WO2025151671A1 - Compositions and methods for treating diet-induced obesity and associated metabolic disorders - Google Patents

Abstract

Embodiments pertain to a composition suitable for use in treating or preventing a metabolic disorder in a subject. The composition may include a therapeutically effective amount of indacaterol, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof. The composition may also include at least one glucagon-like peptide- 1 (GLP-1) agonist. Additional embodiments pertain to methods of treating or preventing a metabolic disorder in a subject by administering to the subject a composition of the present disclosure.

Description

TITLE

COMPOSITIONS AND METHODS FOR TREATING DIET-INDUCED OBESITY AND ASSOCIATED METABOLIC DISORDERS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63/620,574, filed on January 12, 2024. The entirety of the aforementioned application is incorporated herein by reference.

BACKGROUND

[0002] A need exists to develop novel therapeutic strategies for treating or preventing obesity and related metabolic disorders. Numerous embodiments of the present disclosure aim to address the aforementioned need.

SUMMARY

[0003] In some embodiments, the present disclosure pertains to a composition suitable for use in treating or preventing a metabolic disorder in a subject. In some embodiments, the composition includes a therapeutically effective amount of at least one compound. In some embodiments, the compound includes, without limitation, indacaterol, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof. In some embodiments, the compound also includes at least one glucagon-like peptide- 1 (GLP-1) agonist. In some embodiments, the GLP-1 agonist includes, without limitation, semaglutide, exenatide, liraglutide, albiglutide, dulaglutide, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof.

[0004] Additional embodiments of the present disclosure pertain to methods of treating or preventing a metabolic disorder in a subject by administering to the subject a composition of the present disclosure. The compositions of the present disclosure may be utilized to treat or prevent various metabolic disorders. For instance, in some embodiments, the metabolic disorder to be treated or prevented includes, without limitation, obesity, diet-induced obesity, type 2 diabetes (T2D), diet-induced hyperglycemia, cardiovascular disease, hyperlipidemia, hepatic lipidosis, liver fibrosis, metabolic syndrome, insulin resistance, or combinations thereof.

[0005] The compositions of the present disclosure may be administered to various subjects. For instance, in some embodiments, the subject is a human being. In some embodiments, the subject is an obese subject. In some embodiments, the obese subject has a body mass index of more than 30 (BMI >30).

DESCRIPTION OF THE DRAWINGS

[0006] FIGS. 1A-1H illustrate the identification of FDA approved drugs targeting the catalytic binding domain of hMLYCD. FIG. 1A shows crystal structures of hMLYCD (PDB ID:2YGW) highlighting the catalytic dyad of S329 (Magenta) and H423 (Blue). This is supported with key amino acids residues needed for binding and highlighted as grey spheres including L253, F288, N421, F422, and Y456. This was downloaded and processed using Pymol. FIG. IB provides a schematic flow for HTVS using FRED from OpenEye molecular modeling software to elucidate 15 drugs showing three binding modes within the catalytic binding domain of hMLYCD. FIGS. 1C-1E show representative drugs (Pemetrexed, Losartan, and Indacaterol) representing binding mode 1-3 generated by Vida, where the three drugs showed potential hydrophobic-hydrophobic interaction supported with hydrogen bonds (dashed green lines). FIGS. 1F-1H show IC50 profiling of Pemetrexed, Losartan, and Indacaterol against MLYCD activity in a dose-dependent manner, monitored with excitation Z340 nm and an emission Z460 nm. The activity (%) was calculated based on control. (n=3 independent trials, data are presented; mean ± SEM). [0007] FIGS. 2A-2G illustrate that indacaterol binds to hMLYCD. FIG. 2A shows the catalytic binding domain of MLYCD in the absence and presence of Indacaterol (Purple) featuring the catalytic dyad S329 (magenta) and H423 (blue). FIG. 2B shows IC50 profiling of Indacaterol against MLYCD activity in a dose- and time-dependent manner. The activity (%) was calculated based on control (n=3 independent replicates, data are presented; mean ± SEM). FIG. 2C shows a thermal shift assay of MLYCD in the presence of Indacaterol and CBM-301940. FIGS. 2D-2E show relative Oil Red O staining and quantification for 3T3-L1 fibroblasts stained with different concentrations of indacaterol. FIG. 2F shows Seahorse XFe24 extracellular flux analyzer traces of a glycolysis stress test. FIG. 2G shows quantification of glycolysis, glycolytic capacity, and glycolytic reserves. Statistical analyses were carried out using one-way ANOVA followed by Bonferroni’s post-hoc analysis and unpaired student t-test, where ****p<0.0001, ***p<0.001, **p<0.01, and *p<0.05.

[0008] FIGS. 3A-3M show in vivo evaluation of indacaterol (5 and 10 mg/kg/day, s.c.) administration in the diet-induced obesity mouse model. FIG. 3A provides a schematic for in vivo administration of indacaterol (5 andlO mg/kg/day, s.c.) in a high fat diet (HFD)-induced obesity mouse model. FIG. 3B shows whole-body weight for vehicle and indacaterol mice in HFD- induced obese mice after 6 weeks of administration. FIG. 3C shows total-body weight measurement over 6 weeks of administration of vehicle and indacaterol subcutaneous at 5 and 10 mg/kg/day, s.c. for high fat diet-induced obesity mouse model compared to matched age mice on regular diet. FIGS. 3D-3E show serum total Cholesterol and free fatty acid. FIGS. 3F-3G show a glucose tolerance test and AUC data. FIGS. 3H-3I show an insulin tolerance test and AUC data. FIGS. 3J-3K show fat and lean mass. FIGS. 3L-3M show interscapular brown adipose tissue images using high-resolution FLIR T560 infra-red thermal imaging for vehicle and indacaterol (5 and 10 mg/kg/day, s.c.). Data is presented as mean ± SEM. Statistical analyses were carried out using one-way ANOVA followed by Bonferroni’s post-hoc analysis, where ****p<0.0001, ***p<0.001, **p<0.01, and *p<0.05. [0009] FIGS. 4A-4J show that indacaterol reduces food intake and elevates metabolic rates and thermogenesis in a HFD-induced obesity mouse model. FIG. 4A shows a schematic for metabolic profiling in the DIO mice using indirect calorimetry chamber (n= 4 for each group). FIGS. 4B- 4C show oxygen (VO2) consumption over three days and average (VO2). FIGS. 4D-E show carbon dioxide (VCO2) production over three days and average (CO2). FIGS. 4F-G show RER over three days and average RER. FIGS. 4H-I show heat over three days and average in both dark and light cycles. FIG. 4J shows cumulative food intake. Data is presented as mean ± SEM. Panels in FIGS. 4B-G and 4J were analyzed by one-way ANOVA, followed by Bonferroni’s post- hoc analysis. Panels in FIGS. 4H-I were analyzed by linear mixed models with repeated measures using body composition = 29.295 g as a covariatc where ****p<0.0001, ***p<0.001, **p<0.01, and *p<0.05.

[0010] FIGS. 5A-5O show that indacaterol activates glucose oxidation and insulin signaling while inhibiting MLYCD activity in HFD-obese mouse models. FIG. 5A shows a schematic for in vivo administration of vehicle and indacaterol in HFD-induced obese mice until harvest of snap-frozen livers. FIG. 5B shows immunoblots for vehicle and indacaterol (5 and 10 mg/kg) -treated liver lysates (n=5 independent replicates for each group). FIGS. 5C-5H show densitometry analysis for FASN, CPT1A, CD36, p-AKT, AKT, MLYCD, p-PDH, PDH, and P-actin expression levels. FIG. 51 show immunoblots for vehicle and indacaterol (5 and 10 mg/kg)-treated brown adipose tissue lysate lysates (n=5 independent replicates for each group). FIGS. 5J-O show densitometry analysis for FASN, CPT1A, CD36, p-AKT, AKT, MLYCD, p-PDH, PDH, and P-actin expression levels. Data is presented as mean ± SEM. Statistical analyses were carried out using one-way ANOVA followed by Bonferroni’s post-hoc analysis, where ****p<0.0001, ***p<0.001,

**p<0.01, and *p<0.05.

[0011] FIGS. 6A-6B show safety and pharmacokinetics (PK) profiling of Indacaterol. FIG. 6A show representative images for H&E staining of vehicle and indacaterol (5 and 10 mg/kg/day, s.c.) treated mice (n=3 for each group) over a 50-day treatment period for isolated liver, kidney, and lung tissues. The red box highlights the presence of mild alveolar inflammation for lung tissue treated with 10 mg/kg of indacaterol. FIG. 6B shows PK behavior of indacaterol (5mg/kg, s.c.). Data is presented as mean ±SD (n=3 C57B6/J mice). Where error bars are not visible, they fall within the symbols. The inset shows the first 8 h after administration.

[0012] FIGS. 7A-7M show that indacaterol can reduce whole-body weight and improve glucose homeostasis in HFD-induced obese mice independently in the presence of non-selective pharmacological blocking of P-adrenergic receptor. FIG. 7A shows a fluorescence-based MLYCD assay for selective 2 adrenergic agonists at 10 pM monitored with excitation X340 nm and an emission X460 nm (n=2 independent trials). FIG. 7B shows a schematic for concurrent administration of propranolol (5 mg/kg/day, i.p.) and indacaterol (5 mg/kg/day, sc.) (n=4-5 mice for each group). FIG. 7C shows a whole-body weight for vehicle and indacaterol mice in HFD- induced obese mouse after 50 days. FIG. 7D shows a total-body weight measurement for vehicle, indacaterol (5 mg/kg/day, s.c.), and propranolol (5 mg/kg/day, i.p.) + indacaterol (5 mg/kg/day, s.c.) treated groups. FIGS. 7E-F show a glucose tolerance test and AUC data. FIGS. 7G-H show fat and lean mass. FIG. 71 shows food intake. FIG. 7J shows oxygen (VO2) consumption in the dark and light cycles. FIG. 7K shows carbon dioxide (VCO2) production in the dark and light cycles. FIGS. L-M show interscapular brown adipose tissue images using high-resolution FLIR T560 infra-red thermal imaging for vehicle, indacaterol (5 mg/kg/day, s.c.), and propranolol (5 mg/kg/day, i.p.) + indacaterol (5 mg/kg/day, s.c.). Data is presented as mean ± SEM. Statistical analyses were carried out using one-way ANOVA followed by Bonferroni’s post-hoc analysis, where ****p<0.0001, ***p<0.001, **p<0.01, and *p<0.05. [0013] FIGS. 8A-8E show RNA-Seq for isolated livers from indacaterol- and vehicle-treated HFD-induced obese mice. FIG. 8A shows Venn diagrams, which show the common genes of two cohorts (Vehicle vs Indacaterol-treated livers, normalized to Ctrl). FIG. 8B shows a heat map for representative downregulated genes induced by indacaterol administration. FIG. 8C shows GO of the downregulated pathways assigning fatty acid metabolism, carbohydrate/glucose metabolism, and insulin regulation. FIG. 8D shows a heat map for representative upregulated genes induced by indacaterol administration. FIG. 8E shows GO of the upregulated pathways assigning fatty acid metabolism, carbohydrate/glucose metabolism, and insulin regulation.

[0014] FIGS. 9A-9C show transcriptomic signature for Indacaterol in metabolic reprogramming by inhibiting fatty acid synthesis/oxidation and activating glycolysis/glucose oxidation. FIG. 9A shows a graphical presentation of expressed genes in fatty acid synthesis pathway in indacaterol- treated mice. FIG. 9B shows graphical presentation of expressed genes in fatty acid oxidation pathway in indacaterol-treated mice. FIG. 9C shows graphical presentation of expressed genes in glycolysis and glucose oxidation in indacaterol-treated mice. RNA-seq determined differential gene expressions for isolated livers from indacaterol-treated HFD-induced obese mice.

[0015] FIGS. 10A-10L show in vivo evaluation of indacaterol (5 mg/kg/day, s.c.) and semaglutide (10 nmol/kg, s.c.) administration in the diet-induced obesity mouse model. FIG. 10A shows whole-body weight for the vehicle, indacaterol, and semaglutide mice in HFD-induced obese mice after 35 days of administration. FIG. 10B shows total-body weight measurement over 35 days administration of vehicle, indacaterol, and semaglutide subcutaneous. FIGS. 10C-D show lean and fat mass. FIGS. 10E-F show cumulative food intake. FIGS. 10G-H show oxygen (VO2) consumption over three days and average (VO2). FIGS. 101- J show carbon dioxide (VCO2) production over three days and average (CO2). FIGS. 10K-L show total energy expenditure over three days and average in both dark and light cycles. Data is presented as mean ± SEM. Statistical analyses were carried out using one-way ANOVA followed by Bonferroni’s post-hoc analysis, where ****p<0.0001, ***p<0.001, **p<0.01, and *p<0.05. [0016] FIGS. 11A-11C show in vivo evaluation of dual administration of 1.6 mg/kg indacaterol and lOnmol/kg semaglutide in HFD-induced obese mice. FIG. 11A shows total-body weight measurement over 30 days administration of vehicle and dual administration of indacaterol and semaglutide subcutaneous. FIGS. 11B-11C show lean and fat mass. Data is presented as mean ± SEM. Statistical analyses were carried out using Welsh’s t-test analysis, where ****p<0.0001, ***p<0.001, **p<0.01, and *p<0.05.

DETAILED DESCRIPTION

[0017] It is to be understood that both the foregoing general description and the following detailed description are illustrative and explanatory, and are not restrictive of the subject matter, as claimed. In this application, the use of the singular includes the plural, the word “a” or “an” means “at least one”, and the use of “or” means “and/or”, unless specifically stated otherwise. Furthermore, the use of the term “including”, as well as other forms, such as “includes” and “included”, is not limiting. Also, terms such as “element” or “component” encompass both elements or components comprising one unit and elements or components that include more than one unit unless specifically stated otherwise.

[0018] The section headings used herein are for organizational purposes and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including, but not limited to, patents, patent applications, articles, books, and treatises, are hereby expressly incorporated herein by reference in their entirety for any purpose. In the event that one or more of the incorporated literature and similar materials defines a term in a manner that contradicts the definition of that term in this application, this application controls.

[0019] Obesity and related metabolic disorders represent public health concerns. In particular, obesity is a global health socioeconomic problem with a prevalence of 41%, leading to a significant mortality rate of four million cases annually worldwide. By 2030, around 50% of Americans will suffer from obesity and associated complications. [0020] Obesity is linked to several co -morbidities, including hyperglycemia, type 2 diabetes (T2D), dyslipidemia, cardiovascular disease, and heart failure. Obesity usually occurs due to high- calorie intake than is burned by exercise or normal daily activities, leading to increased body mass index and abnormal or massive fat accumulation.

[0021] One of the key factors in obesity is leptin resistance, which causes fat to accumulate in nonadipose tissues such as the pancreas and skeletal muscles. This accumulation results in increased fatty acid oxidation and free fatty acid production, which induces peripheral insulin resistance. This process leads to lipotoxicity, reduced glucose oxidation, and the development of T2D.

[0022] The current non-surgical therapeutic interventions for obesity are limited to (1) dietary caloric restrictions, (2) administration of Orlistat that can inhibit pancreatic lipases to prevent the absorption of ingested fats up to 32% to be excreted in the feces, leading to steatorrhea, (3) administration of a combination of Naltrexone (opioid-based antagonist) and bupropion (antidepressant) to suppress appetite and hunger centers in the hypothalamus with severe adverse effects, (4) administration of centrally acting serotonin agonists and combinations of centrally acting appetite suppressing and antiepileptic drugs, and more recently, (5) glucagon-like peptide- 1 (GLP-1) receptor agonists (GLP-1 RA). GLP-1 agonists, such as Liraglutide (Saxenda®), have shown significant potential for inducing dose-dependent weight loss in patients with T2D. However, there are several reports of adverse effects associated with the use of GLP-1 analogs. These adverse effects include chronic intestinal pseudo-obstruction (CIP) and loss of lean body mass.

[0023] The American Diabetes Association (ADA) has warned about the risks of compounded GLP-1 RA and dual GIP/GLP-1 RA products, which are not FDA-approved. These medications have been linked to dosing errors and adverse events. Additionally, counterfeit versions have entered the U.S. market and are being sold online by unregulated sources, posing serious safety risks due to questionable quality and efficacy. Therefore, there is still a critical need to develop novel therapeutic strategics while tackling integrated molecular targets contributing to the progression of obesity and associated metabolic disorders. [0024] During fatty acid synthesis, malonyl-CoA is produced by acetyl-CoA carboxylase. Subsequently, malonyl-CoA is metabolized by fatty acid synthase (FASN) and incorporated into long-chain fatty acids. The enzyme malonyl-CoA decarboxylase (MLYCD) catalyzes the decarboxylation of malonyl-CoA to produce acetyl-CoA, which stimulates the mitochondrial uptake of free fatty acids for [3-oxidation. This relieves the malonyl-CoA-mediated inhibition of carnitine palmitoyl transferase (CPT1), the rate-limiting enzyme in fatty acid oxidation. Overexpression of MLYCD in the skeletal muscles for mouse models resulted in a decline in malonyl-CoA levels, exacerbating insulin resistance and inhibiting insulin signaling for high-fat diet (HFD)-induced obese mouse models. Previous studies showed that when MLYCD was ovcrcxprcsscd in the mediobasal hypothalamus of rats, it lowered malonyl-CoA levels and increased food intake.

[0025] MLYCD inhibitors centrally increased malonyl-CoA levels in the hypothalamus, reduced food intake, and induced hypophagia. Several studies showed that MLYCD knock-out (MLYCD'' ) in cell lines lead to inhibition of fatty acid oxidation and induction of tricarboxylic acid (TCA) cycle alteration mediated by accumulation of Malonyl-CoA. Additionally, MLYCD knock-out (MLYCD^-) adult mice with ischemic heart diseases can inhibit fatty acid oxidation through malonyl-CoA inhibitory effect for CPT1 to shift energy metabolism in cardiometabolic diseases and promote glucose oxidation. Additionally, the inhibition of MLYCD is associated with improved insulin sensitivity and decreased inflammatory response in obese animal models.

[0026] MLYCD inhibitors managed to inhibit fatty acid oxidation and stimulate cardiac glucose oxidation, thus protecting the heart from ischemic injury and improving cardiac function post- myocardial infarction. Conversely, MLYCD knock-out (MLYCD''') mice had lower survival rates and health issues around and before weaning, including lower body weight, increased body fat, liver problems, and heart issues. However, these problems improved as the mice aged, especially when their diet changed from high-fat to standard. Despite such convincing evidence, there is limited progress in the development of therapeutic drugs that specifically target MLYCD. [0027] As such, a need exists to develop novel therapeutic strategies for treating or preventing obesity and related metabolic disorders. Numerous embodiments of the present disclosure aim to address the aforementioned need.

[0028] Compositions

[0029] In some embodiments, the present disclosure pertains to a composition. In some embodiments, the composition is suitable for use in treating or preventing a metabolic disorder in a subject. In some embodiments, the composition includes a therapeutically effective amount of at least one compound. In some embodiments, the compound includes, without limitation, indacaterol, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof.

[0030] In some embodiments, the compound includes indacaterol. In some embodiments, the indacaterol includes: pharmaceutically acceptable salt thereof, a pharmaceutically acceptable hydrate thereof, or combinations thereof. In some embodiments, the indacaterol includes the following structure: [0031] In some embodiments, the compound also includes at least one glucagon-like peptide- 1 (GLP-1) agonist. In some embodiments, the GLP-1 agonist includes, without limitation, semaglutide, exenatide, liraglutide, albiglutide, dulaglutide, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof. In some embodiments, the GLP-1 agonist includes semaglutide.

[0032] In some embodiments, the GLP-1 agonist is in the same composition as the indacaterol. In some embodiments, the GLP-1 agonist is in a composition separate from the indacaterol.

[0033] In some embodiments, the compositions of the present disclosure also include at least one solubilizing agent. In some embodiments, the solubilizing agent includes, without limitation, polyethylene glycol, glycerin, propylene glycol, ethanol, sorbitol, polyoxyethylated glycerides, polyoxyethylated oleic glycerides, polysorbates, sorbitan monooleate, hydroxypropyl-beta- cyclodextrin (HPCD), polyoxyl 40 hydrogenated castor oil, polyoxyl hydroxystearates, or combinations thereof.

[0034] Methods of treating or preventing a metabolic disorder in a sub ject

[0035] Additional embodiments of the present disclosure pertain to methods of treating or preventing a metabolic disorder in a subject by administering to the subject a composition of the present disclosure. As set forth in more detail herein, the methods of the present disclosure can have numerous embodiments.

[0036] Administration.

[0037] The compositions of the present disclosure may be administered to subjects in various manners. For instance, in some embodiments, the administration occurs by a method that includes, without limitation, oral administration, inhalation, subcutaneous administration, intravenous administration, intraperitoneal administration, intramuscular administration, intrathecal injection, intra-articular administration, topical administration, central administration, peripheral administration, aerosol-based administration, nasal administration, transmucosal administration, transdermal administration, parenteral administration, intravenous administration, or combinations thereof. In some embodiments, the administration occurs by subcutaneous administration. In some embodiments, the administration occurs by intravenous administration.

[0038] The compounds in the compositions of the present disclosure may be administered at various amounts. For instance, in some embodiments, indacaterol in a composition of the present disclosure is administered at a concentration of less than about 100 mg per kg of body weight. In some embodiments, indacaterol in a composition of the present disclosure is administered at a concentration of less than about 50 mg per kg of body weight. In some embodiments, indacaterol in a composition of the present disclosure is administered at a concentration of less than about 25 mg per kg of body weight. In some embodiments, indacaterol in a composition of the present disclosure is administered at a concentration of less than about 15 mg per kg of body weight. In some embodiments, indacaterol in a composition of the present disclosure is administered subcutaneously at a concentration of about 5 and 10 mg per kg of body weight. In some embodiments, indacaterol in a composition of the present disclosure is administered subcutaneously at concentrations of 5 and 10 mg per kg of body weight. In some embodiments, indacaterol in a composition of the present disclosure is administered subcutaneously at a concentration of 1.6 mg/kg of body weight.

[0039] In some embodiments, the indacaterol is co-administered with at least one glucagon-like peptide-1 (GLP-1) agonist. In some embodiments, the GLP-1 agonist is in the same composition as the indacaterol. In some embodiments, the GLP-1 agonist is in a composition separate from the indacaterol. [0040] In some embodiments, a GLP-1 agonist (e.g., Semaglutide) is administered at a concentration of at least about 10 nmol per kg of body weight. In some embodiments, a GLP-1 agonist is administered at a concentration of at least about 50 nmol per kg of body weight. In some embodiments, a GLP-1 agonist (e.g., Semaglutide) is administered at a concentration of at least about 100 nmol per kg of body weight. In some embodiments, a GLP-1 agonist (e.g., Semaglutide) is administered subcutaneously at a concentration of 10 nmol per kg of body weight. In some embodiments, a GLP-1 agonist is administered at a concentration of 10 nmol per kg of body weight. In some embodiments, a GLP-1 agonist (e.g., Semaglutide) is administered at a concentration of 10 nmol per kg of body weight.

[0041] In some embodiments, a GLP-1 agonist (e.g., Semaglutide) is administered subcutaneously at a concentration of 10 nmol per kg of body weight results in weight loss, reduced fat mass and reduced lean mass. In some embodiments, concurrent administration of 10 nmol per kg of Glp-1 agonist (e.g., Semaglutide) and Indacaterol at a concentration of 1.6 mg per kg body weight results in reduced body weight, reduced fat mass, and preserved lean muscle mass.

[0042] Metabolic disorders

[0043] The compositions of the present disclosure may be utilized to treat or prevent various metabolic disorders. For instance, in some embodiments, the metabolic disorder to be treated or prevented includes, without limitation, obesity, diet-induced obesity, type 2 diabetes (T2D), diet- induced hyperglycemia, cardiovascular disease, hyperlipidemia, hepatic lipidosis, liver fibrosis, metabolic syndrome, insulin resistance, or combinations thereof.

[0044] In some embodiments, the metabolic disorder to be treated or prevented includes obesity. In some embodiments, the metabolic disorder to be treated or prevented includes severe obesity. Severe obesity can represent serious introduction to associated comorbidities, including insulin resistance, T2D, coronary artery diseases, renal disorders, and colon cancer. Such conditions are the main underlying causes for elevated levels of morbidity and mortality worldwide.

[0045] Subjects [0046] The compositions of the present disclosure may be administered to various subjects. For instance, in some embodiments, the subject is a human being. In some embodiments, the subject is an obese subject. In some embodiments, the obese subject has a body mass index of more than 30 (BMI >30). In some embodiments, the subject is diagnosed with type 2 diabetes. In some embodiments, the subject is suffering from obesity, diet-induced obesity, type 2 diabetes (T2D), diet-induced hyperglycemia, cardiovascular disease, hyperlipidemia, hepatic lipidosis, liver fibrosis, metabolic syndrome, insulin resistance, or combinations thereof.

[0047] In some embodiments, the subject is a non-human mammal. In some embodiments, the non-human mammal includes, without limitation, a horse, a rabbit, a mouse, a rat, a pig, a sheep, a cow, a dog, or a cat. In some embodiments, the non-human mammal is a domestic animal, such as a dog or a cat.

[0048] The compositions of the present disclosure may have various therapeutic effects on subjects. For instance, in some embodiments, the administration of the composition results in weight loss, restoration of glucose homeostasis, improvement in insulin sensitivity, reduced total serum cholesterol, reduced free fatty acid levels, reduced fatty liver, reduced food intake, enhanced metabolic rate, or combinations thereof. In some embodiments, the administration of the composition results in weight loss.

[0049] Additional Embodiments

[0050] Reference will now be made to more specific embodiments of the present disclosure and experimental results that provide support for such embodiments. However, Applicant notes that the disclosure below is for illustrative purposes only and is not intended to limit the scope of the claimed subject matter in any way.

[00 1 ] Example 1. Indacaterol induces metabolic reprogramming in obesity [0052] Obesity is a common debilitating condition that poses a significant financial burden to global healthcare. The dysfunction of key regulatory enzymes of fatty acid synthesis and oxidation predisposes to obesity and associated metabolic diseases. Malonyl-CoA is known for its metabolic regulatory role in fatty acid synthesis, and it is decarboxylated by malonyl-CoA decarboxylase (MLYCD) to produce acetyl-CoA.

[0053] Although pharmacological targeting of MLYCD could be a viable path to treat obesity, there are no FDA-approved drugs targeting MLYCD. Therefore, Applicant performed an in-silico screen to identify FDA-approved drugs that inhibit MLYCD catalytic activity. Applicant’s screen yielded several promising candidates based on binding profiles as well as safety and side effect profiles.

[0054] Applicant screened the top 15 hits for their inhibitory profiles against MLYCD activity. Indacaterol, a selective P2 adrenergic receptor agonist, showed inhibitory profiles against MLYCD catalytic activity in a time-dependent manner. This was associated with a significant change in the melting curves in MLYCD when incubated with indacaterol. Subcutaneous administration of indacaterol to high-fat diet-induced obese mice reduced food intake, enhanced thermogenesis, induced weight loss with preservation of lean mass, restored glucose homeostasis, improved insulin sensitivity, and reduced total serum cholesterol as well as free fatty acid levels, independent of pharmacological inhibition of P-adrenergic receptor, and with no toxicity.

[0055] Notably, RNA-Seq and immunoblotting of key markers revealed that indacaterol results in metabolic reprogramming by inducing a metabolic switch from fatty acid utilization to glucose oxidation. Collectively, these results identify indacaterol as an FDA-approved drug with therapeutic potential for the treatment of obesity and associated metabolic diseases.

[0056] Example LI. Structural basis and small molecules for targeting MLYCD [0057] The crystal structure of apo hMLYCD (PDB ID: 2YGW) was elucidated to provide structural insights for the binding mechanism of malonyl-CoA (PMID 23791943). However, the crystal structures of hMLYCD with malonyl-CoA or acetyl-CoA had not been successfully resolved. MLYCD catalytic domain shares structural homology with GCN5-related N- acetyltransferase superfamily with a conserved His-Ser/Thr catalytic dyad induced by acetyl-CoA. However, there are limited examples of MLYCD inhibitors. For example, the MLYCD inhibitors CBM-3000864 and CBM-301940 have been shown to block malonyl-CoA degradation and decrease fatty acid oxidation while accelerating glucose oxidation without changing glycolysis rates to protect ischemic hearts in demand-induced ischemia in pigs. This resulted in a metabolic switch in the energy substrate utilization to glucose as the primary energy source in the heart.

[0058] Later, novel heteroaryl-based analogs targeting MLYCD, including isoxazole, imidazole, pyridyl, and thiazole-based derivatives coupled with bis-trifluoromethyl moiety to offer promising hits in the nanomolar range. However, preclinical testing was not conducted on most of these compounds. Although MLYCD is a druggable candidate for preventing and treating obesity and metabolic disorders, there are no clinically proven nor FDA-approved MLYCD inhibitors. Therefore, Applicant’s objectives are to screen FDA-approved drugs that target MLYCD, characterize the binding profile of the most promising drugs against MLYCD, assess the in vivo potency of the drug on glucose and lipid metabolism, and determine the preclinical effects of chronic systemic administration of the drug on energy balance and body composition.

[0059] Example 1.2. In silico molecular modeling of FDA-approved drugs that can target the catalytic dyad of MLYCD

[0060] Applicant leveraged the resolved X-ray crystal structure of hMLYCD (PDB ID: 2YGW) for high-through output virtual screening (HTVS) via docking of the energy-minimized FDA- approved drug library using FRED from OpenEye© molecular modeling software. Applicant’s preliminary structural analysis revealed three binding modes for potential 15 hits to the catalytic bind domain of MLYCD (FIG. IB). [0061] Applicant’s top hits were categorized based on their binding mode profiles, where binding mode (1) included Pemetrexed, Pimozide, Dexlanzoprazole, Delavirdine, Pazopanib, Pantoprazole, Guanfacine, and Paroxetine (FIGS. 1A and 1C). Binding mode (2) involved three angiotensin II receptor blockers: Losartan, Irbesartan, and Valsartan (FIGS. ID and IB). Binding mode (3) included Indacaterol, Levomefolic acid, Folinic acid, and Calciol (FIGS. IE and 1C). Applicant’s structural analysis suggested that binding mode (1) showed better occupancy for the hydrophobic pocket of the catalytic binding domain of MLYCD supported by a network of hydrogen bonds with S292, T305, and F422. Binding mode (2) showed a similar binding pattern within the same catalytic pocket, lacking further extended occupancy to fill the hydrophobic pocket represented by F422, N426, and T305. Applicant extended docking simulations to assign S329 and H423 as constraints while HTVS for the FDA library. This elucidated the structural insights shown by binding mode (3) considering hydrophobic-hydrophobic interactions supported with hydrogen bonding represented by S292, P332, H423, and D438.

[0062] Example 1,3. Indacaterol can inhibit MLYCD activity

[0063] Next, Applicant screened the top 15 hits against MLYCD catalytic activity at 10 pM using fluorescence-based MLYCD assays by monitoring acetyl-CoA formation using the malate dehydrogenase (MD)/citrate synthase (CS) coupling system, as previously described. Next, Applicant conducted IC50 profiling for pemetrexed (1.73 ± 0.28 pM), losartan (9.78 ± 2.01 pM), and indacaterol (4.27 ± 1.31 pM) and showed significant inhibitory profiles for MLYCD compared with the control (1% DMSO) (FIGS. 1F-1H).

[0064] Indacaterol, a selective 02 adrenergic receptor agonist, inhibited MLYCD catalytic activity in a dose-dependent manner at different residence times at IC504.27 ± 1.31 pM (30 mins), 0.54 ± 0.173 pM (Ihr), and 0.086 ± 0.01 pM (2hr) (FIG. 2B). Applicant’s thermal shift assay (TSA) showed that the melting curve behavior for MLYCD and indacaterol (ATm = 8.30 ± 1.04 °C), supported by the behavior of CBM-301940 (positive control, MLYCD inhibitor) to show (ATm = 1.50 ± 0.50 °C) (FIG. 2C). This suggests that indacaterol can induce significant changes upon binding to MLYCD, compared to vehicle and positive control. [0065] Collectively, these results support the potential notion that indacaterol directly interacts with purified hMLYCD. Applicant’s in vitro studies suggested that indacaterol inhibits lipid accumulation after differentiation to adipocytes in the embryonic mouse 3T3-L1 fibroblast using Oil Red O staining in a dose-dependent manner compared to vehicle- treated cells (FIGS. 2D-2E). Applicant performed a seahorse glycolysis stress test on differentiated 3T3-L1 cells that showed a significant elevation of glycolysis, glycolytic capacity, and reverse after treating cells with indacaterol at a concentration of (IpM) compared to vehicle (FIGS. 2F-2G).

[0066] Example 1.4. Indacaterol promotes weight loss, improves whole -blood glucose levels, mitigates insulin resistance, reduces food intake, and elevates metabolic rates in a high-fat diet (HFD)-induced obesity mouse model

[0067] Next, Indacaterol was administrated subcutaneously (s.c.) at two sub-maximal tolerated doses (5 and 10 mg/kg/day) in DIO mice in the HFD-induced obesity C57BL/6J mouse model for six weeks. The indacaterol-treated groups significantly reduced body weight, total serum cholesterol, and free fatty acid levels compared to the vehicle-treated group associated with improving glucose homeostasis and insulin sensitivity after 4 weeks of daily injections (FIGS. 3A-I). Body composition analysis at the end of the study showed that indacaterol treatment significantly reduced fat mass while preserving lean mass (FIGS. 3J-K). The Applicant also recorded the interscapular brown adipose tissue (iBAT) temperature using high-resolution FLIR infrared thermal imaging camera that showed elevation in the BAT temperature in the treated group compared to the control group indicating increased thermogenesis and metabolism (FIGS. 3L-M).

[0068] Applicant has used food intake monitoring systems and energy expenditure chambers to understand the mechanism of body weight loss associated with indacaterol administration. The treated mice groups showed elevated VO2 consumption and VCO2 production rates during the dark and light cycles, indicating increased metabolic rates (FIGS. 4B-4E). Regarding the respiratory exchange rate RER, the treated group showed a significant elevation in both dark and light cycles (FIGS. 4F-G). This indicates a switch in the substrate utilization preference to glucose utilization instead of fatty acid oxidation. Interestingly, indacaterol-treated mice showed a significant elevation of energy expenditure during the dark cycle, indicating increased whole-body thermogenesis (FIGS. 4H-I). That was coupled with a 23% reduction in food intake in both dark and light cycles (FIG. 4J). Next, Applicant harvested the liver of indacaterol and vehicle-treated cohorts to show the significant reduction of oil red o staining for indacaterol-treated liver, suggesting the ability of indacaterol to inhibit lipid accumulation.

[0069] Example 1.5. Indacaterol induces fatty acid utilization switch towards glucose oxidation and activates insulin signaling in vivo

[0070] To gain insights into signaling mechanisms of indacaterol-induced improvements in glucose clearance, liver and brown adipose tissues were harvested and lysed for western blotting analysis for vehicle- and indacaterol-treated cohorts (FIGS. 5A, 5B, and 51). The mitochondrial pyruvate dehydrogenase (PDH) is a central enzyme regulating glucose utilization relative to fatty acids for energy homeostasis. Previously, cardiac-specific deletion of pyruvate dehydrogenase kinase 4 (PDK4) resulted in the activation of PDH activity to induce mitochondrial substrate switch from fatty acids to glucose utilization.

[0071] Indacaterol-treated liver lysates inhibited fatty acid synthetase (FASN), CPT1A1, CD36, and p-PDH without affecting total PDH levels compared to vehicle-treated liver lysate for HFD- obese mice, showing the activation of PDH activity and inhibition of CD36, to suggest the shift of TCA cycle metabolism from fatty acid to glucose oxidation in the isolated livers (FIGS. 5C-F) and brown adipose tissues (FIGS. 5J-M). Herein, Applicant shows the ability of indacaterol to increase in the AKT phosphorylated signal (p-AKT), suggesting the insulin signaling activation in the isolated livers (FIG. 5G) and brown adipose tissue (FIG. 5N). This was associated with a significant reduction of MLY CD for indacaterol-treated liver lysates (FIG. 5H) and brown adipose tissues (FIG. 50).

[0072] Example 1.6. Safety and pharmacokinetics (PK) profiling of Indacaterol

[0073] Next, Applicant identified the toxicological profiles for subcutaneous administration of vehicle and indacaterol (5 and 10 mg/kg/day) over a 50-day treatment period through isolation of liver, kidney, and lung tissue for histopathological analysis to account for necrosis, inflammation, or neoplasia. The two dose levels showed no pronounced morphological toxicity in the isolated tissues. However, the 10 mg/kg dose showed mild (5-10%) perivascular and alveolar inflammation in the lung tissue (FIG. 6A). Hence, Applicant pursued subsequent experiments at a 5 mg/kg dose level.

[0074] Applicant analyzed the indacaterol plasma concentrations after a single subcutaneous dose of 5 mg/kg up to 144 h in C57B6/J mice (FIG. 6B). Non-compartmental analysis of the concentration-time course generated the parameters. Cmax (132.5 ± 35.6 ng/ml, 0.33 pM) observed at Tmax (Ihr) is higher than IC50 of indacaterol against MLYCD after 2 hrs of pre-incubation at 0.08 pM in vitro. Based on the PK profile after single-dose subcutaneous administration, no significant accumulation of plasma concentrations at steady state would be expected upon longterm once-daily dosing, despite the relatively long half-life of about 80 h observed in the terminal phase (72h and later). This follows from the very low concentration values in that terminal phase, which arc on the order of 1% or less of Cmax. [0075] Example 1,7, Indacaterol reduces whole-body weight and improves glucose homeostasis in HFD-induced obese mice independently in the presence of pharmacological inhibition of 0- adrenergic receptor

[0076] Next, Applicant addressed a mechanistic question if indacaterol’ s phenotype is due to the P2-adrenergic agonistic activity. In humans, p2-adrenergic agonists increase resting metabolic rate by 10-50%, increase muscle mass, and reduce fat mass in a dose-dependent manner. However, recent reports highlighted the limitation of salbutamol to induce weight loss in humans, which suggest that the main mechanism of action is due to p2-adrenergic agonistic activity. The results showed that salbutamol stimulates the glucose uptake by brown adipose tissue only while it has no effect on glucose uptake by skeletal muscles.

[0077] This prompted Applicant to screen other members for p2 adrenergic receptor agonists (Salbutamol, formoterol, albuterol, terbutaline, and olodaterol) along with indacaterol at 10 pM using fluorescence-based MLYCD assay. Only Indacaterol inhibited MLYCD activity compared to P2 receptor agonists. This provides evidence that Indacaterol can contribute to whole-body weight loss due to MLYCD inhibition (FIG. 7B).

[0078] Suppose the metabolic effects of indacaterol are mediated by P-adrenergic signaling. In that case, administration of a non-selective P-adrenergic blocker is expected to reverse or attenuate the effects of indacaterol on energy balance and glucose clearance. To test this, Applicant administered propranolol (5 mg/kg/day, i.p; non-selective P-blocker) before administration of indacaterol (5 mg/kg/day, s.c.). Applicant found that indacaterol-treated mice and Propranolol+ indacaterol-treated mice had similar reductions in body weight, fat mass, and food intake, comparable increases in metabolism rate and iBAT temperature, and similar improvements in glucose clearance (FIGS. 7C-M). [0079] These findings suggest that indacaterol can reduce whole-body weight at 5 mg/kg and improve energy balance and glycemic control in an independent p2-adrenergic sympathetic activity. This experiment was repeated at a higher dose of indacaterol, where propranolol (5 mg/kg/day, i.p) was administered, followed by indacaterol (10 mg/kg/day, s.c.). As with the low dose study, the high dose of indacaterol and indacaterol+ propranolol similarly reduced body weight and food intake, increased metabolic rates with elevated VO2 consumption and VCO2 production rates, increased IBAT temperature, and improved glucose clearance. Together, these results confirm that indacaterol's weight loss, thermogenic effect, and glycemic improvements are independent of p2 adrenergic sympathetic activity.

[0080] Example 1.8. Hepatic transcriptomic signature of indacaterol treated obese mice

[0081] Applicant performed RNA-sequencing (RNA-seq) analysis on isolated livers of indacaterol and vehicle-treated HFD-induced obese mice to understand the differentially expressed genes and associated regulated pathways (FIG. 8A). The RNA-Seq showed that indacaterol downregulated Mlycd, Cptla, Fasn, Acly, Pdkl, Fabp2, Aldob, and Acaalb (FIG. 8B). Gene ontology (GO) analysis of the downregulated genes showed enrichment in several pathways including fatty acid metabolism (fatty acid metabolic process, fatty acid biosynthetic process, fatty acid beta-oxidation, acetyl-CoA metabolic process), carbohydrate/glucose metabolism (glycolytic process, pyruvate biosynthetic process, glycogen metabolic process, regulation of carbohydrates/glucose metabolic process), and insulin regulation (negative regulation of insulin secretion) (FIG. 8C). Reciprocally, the RNA-Seq showed that indacaterol upregulated Adipoq, Lipe, Lepr, Fabp4, Pkm, UCP1, Pfkm, and Irs2 (FIG. 8D).

[0082] GO analysis of the upregulated genes showed enrichment in several pathways of fatty acid metabolism (brown fat cell/ fat cell differentiation), carbohydrates/glucose metabolism (glucose/carbohydrate transmembrane transport, glucose binding/homeostasis), and insulin regulation (positive regulation of insulin secretion) (FIG. 8E). Collectively, indacaterol showed downregulation for the key players in the fatty acid oxidation and upregulation of all the key players in glucose oxidation (FIGS. 9A-9C). This was further validated by KEGG pathway mapping to show the enrichment analysis for several molecular pathways, including upregulation of ECM-receptor interaction, glycolysis, insulin secretion, glucagon signaling pathway, and PPAR signaling pathway, as well as downregulation of fatty acid biosynthesis, fatty acid metabolism, fatty acid degradation, and regulation of lipolysis in adipocytes.

[0083] Example 1,9. Discussion

[0084] Despite recent advances in obesity pharmacotherapies, there is still a significant lack of a metabolic reprogramming therapeutic strategy to reverse the progression of obesity and associated metabolic syndrome. Dysfunction of key regulatory enzymes of fatty acid synthesis and oxidation predisposes to obesity and associated metabolic diseases. Different reports showed that overexpression of MLYCD in skeletal muscles resulted in a decrease of malonyl-CoA levels, leading to exacerbation of insulin resistance via the inactivation of insulin signaling. Another study showed that overexpression of MLYCD in the hypothalamus increased food intake and total body weight. Global knock-out MLYCD mouse models have been validated for metabolic switching towards glucose utilization and improvement of cardiac function in ischemic reperfusion injury mouse models and myocardial infarction. These layers of evidence support a key role for MLYCD in the pathogenesis of obesity and as a potential therapeutic molecular’ target.

[0085] The X-ray crystal structure for MLYCD, showing the catalytic mechanism for malonyl- CoA controlled by the catalytic dyad of His423 and Ser329 for hMLYCD. These findings support the rationale of identifying FDA-approved drugs targeting the catalytic binding domain of hMLYCD via structure-based drug repurposing. [0086] In this Example, Applicant identified novel druggable recognition sites on hMLYCD via HTVS, and screened the top 15 candidates against hMLYCD using a fluorescence-based MLYCD assay, and identified three drugs (Indacaterol, Pemetrexed, and Losartan) based on their IC50 inhibition of MLYCD activity. Given that indacaterol is a long-acting 02 adrenergic receptor agonist that has been approved for treatment of asthma and coronary obstructive pulmonary disease (COPD), it was selected for further testing based on its safety profile and broad therapeutic index.

[0087] Applicant validated that indacaterol managed to bind to the catalytic domain of MLYCD via induction of temperature shift (ATm = 8.30 ± 1.04 °C), compared to CBM-301940 (positive control, MLYCD inhibitor), which showed (ATm = 1.50 ± 0.50 °C) only. Finally, given the promise of these FDA-approved drugs for use in humans, Applicant next investigated their role in the management of obesity using pre-clinical HFD-obese mouse models.

[0088] Based on already identified safety profiles and human equivalent maximum tolerated doses, Applicant evaluated the potential of systemic administration of indacaterol at two doses (5 and 10 mg/kg/day, s.c). Applicant found that indacaterol dose-dependently decreased body weight, which was likely due to reduced food intake associated with concurrent increase in energy expenditure and thermogenesis via modulation of MLYCD activity.

[0089] In contrast, other 02-agonists like clenbuterol and salbutamol do not have the same effect on body weight or food intake. Where, they showed increased food intake and body weight gain in 3 months-old rats with normal-chow diet. The effect of indacaterol can be attributed due to its modulation for G-protein coupled receptor (Gs) activity. However, a recent elegant report showed that Gs activation of intestinal K-cells in HFD-induced obese mice improved glucose tolerance and insulin sensitivity with no effect of body weight, fat/lean mass, and food intake. This suggests that indacaterol showed the anti-obesity phenotype in an independent manner of Gs signaling activation. [0090] The effect of indacaterol on increasing energy expenditure aligns with previous reports showing a significant increase in energy expenditure and physical activity with rest and moderate activity in COPD patients treated with indacaterol for four weeks. Clinical trials on GLP-1 agonists, like semaglutide and liraglutide, indicate that 39-40% of the weight lost with these drugs is lean mass, which is a concern. Importantly, indacaterol reduced fat mass while preserving the lean mass in treated mice, unlike other GLP-1 agonists or p2-agonists like clenbuterol.

[0091] However, Formoterol, an indacaterol-structurally related 2-agonist, showed an increase in lean body mass, and increased soleus and heart mass after chronic administration of 2 mg/kg in rats. Indacaterol significantly improved glucose clearance and insulin resistance, as evidenced by a notable reduction in the AUC for both glucose and insulin tolerance tests. In contrast, previous studies reported that clenbuterol did not affect insulin sensitivity.

[0092] Indacaterol also increased respiratory quotient exchange rates, suggesting that it induces metabolic reprogramming and shifts the preference for systemic substrate use towards glucose oxidation. These findings are consistent with previous studies on increased cardiac glucose oxidation with other MYLCD inhibitors and MYLCD knockout mice.

[0093] Further, Applicant provides evidence that the effects of indacaterol are specific to the inhibition of MLYCD and suppression of hepatic lipogenesis, as indacaterol reduced the relative abundance of MLYCD, FASN, CPT1A1, CD36, and p-PDH signal without affecting total PDH levels, in isolated liver and BAT lysates. An increase in hypothalamic malonyl-CoA levels by central administration of the FASN inhibitor C75 was reported to induce sympathetic nervous system (SNS) activation and upregulation of UCP3 transcript in the skeletal muscle, which was reversed by administration of propranolol, a non-selective p-blocker. Recently, salbutamol, a short-acting P -2- adrenergic receptor agonist, was reported to increase glucose uptake by BAT and increase energy expenditure, which was prevented by propranolol in humans. [0094] Based on these studies, Applicant asked whether indacaterol’s anti-obesity effects are related to its [32- adrenergic receptor agonism. Applicant screened other members for P2 adrenergic receptor agonists (Salbutamol, formoterol, albuterol, terbutaline, and olodaterol) along with indacaterol at 10 pM using a fluorescence-based MLYCD assay. Only Indacaterol inhibited MLYCD activity compared to [32 receptor agonists, suggesting that the weight loss from indacaterol is very likely due to MLYCD inhibition.

[0095] In support of such MLYCD specificity, Applicant showed that propranolol (non-selective P-blocker) did not attenuate or reverse the weight loss, thermogenic, and glycemic improvements, by indacaterol in HFD-induced obese mice. Though the roles of specific P-adrcncrgic receptors in such in vivo effects cannot be confirmed, Applicant’s findings support the notion that indacaterol-induced body weight loss and metabolic and glycemic improvements are partly independent of P-adrcncrgic sympathetic activity. Applicant’s comprehensive RNA-Seq analyses showed downregulation of differentially expressed genes involved in fatty acid utilization such as Mlycd, Cptla, Fasn, Acly, and pdkl for indacaterol-treated livers isolated from HFD-induced obese mice. Reciprocally, indacaterol upregulated hepatic Adipoq, which is involved in regulating glucose homeostasis, lipid metabolism, and insulin sensitivity, and hepatic Lepr that encodes for Leptin receptor protein.

[0096] Applicant also demonstrated the pharmacokinetic profile of indacaterol, and the antiobesity properties that are retained, even at low doses without overt toxicity. Briefly, this Example spans several disciplines to provide druggable recognition sites and identifying therapeutic modalities with established efficacy profiles to target hMLYCD. This was associated with preclinical validation and mechanistic investigation for indacaterol for the first time in HFD- induced obese mouse models. [0097] Applicant believes that subcutaneous systemic administration of indacaterol will offer metabolic reprogramming to induce metabolic switch from fatty acid utilization to glucose oxidation. Collectively, this was associated with significant reduction of whole-body weight, restoration of glucose homeostasis, activation of insulin signaling, and increase of metabolic thermogenesis rates to mitigate obesity and associated metabolic disorders.

[0098] Example 2. Indacaterol in combination with semaglutide induces metabolic reprogramming in obesity

[0099] GLP-1 agonists, such as semaglutide, have shown promise in promoting dose- dependent weight loss in type 2 diabetes (T2D) patients. However, these medications can cause adverse effects like chronic intestinal pseudo-obstruction (CIP) and loss of lean body mass. The American Diabetes Association (ADA) has cautioned against the use of compounded GLP-1 RA and dual G1P/GLP-1 RA products, which are not FDA-approved, due to risks of dosing errors and adverse events. Additionally, counterfeit versions of these medications have entered the U.S. market, being sold online by unregulated sources, posing serious safety risks due to questionable quality and efficacy.

[00100] In this Example, Applicant compares the effects of the FDA-approved drug Indacaterol, as an MLYCD inhibitor, and Semaglutide, a GLP-1 agonist, on high-fat diet (HFD) induced obese mice. The mice received daily treatments of either a vehicle, 5 mg/kg Indacaterol, or 10 nmol/kg Semaglutide subcutaneously for 35 days. Indacaterol and Semaglutide treatments significantly reduced whole-body weight (FIG. 10B), fat mass (FIG. 10D), and food intake (FIGS. 10E and 10F) in mice compared to the vehicle treatment. Interestingly, Indacaterol-treated mice maintained their lean mass, while Semaglutide-treated mice lost about 20% of their lean mass (FIG. 10C). [00101] Indirect calorimetry measurements using energy expenditure clams showed significant O2 consumption (FIGS. 10G and 10H) and CO2 production (FIGS. 10J and 10J) elevations in both indacaterol and semaglutide-treated mice, indicating increased metabolic rates. Indacaterol- treated mice showed a significant increase in total energy expenditure (TEE), while semaglutide- treated mice showed reduced TEE (FIGS. 10K and 10L).

[00102] Next, Applicant addressed the effect of dual administration of 1.6 mg/kg indacaterol and 10 nmol/kg semaglutide in HFD-induced obese mice. This resulted in a significant reduction of total body weight (FIG. 11A) and fat mass (FIG. 11B) while preserving the lean mass (FIG. 11C). This suggests that dual administration of indacaterol and semaglutide can rescue the lean mass loss associated with sole semaglutide administration.

[00103] Without further elaboration, it is believed that one skilled in the art can, using the description herein, utilize the present disclosure to its fullest extent. The embodiments described herein are to be construed as illustrative and not as constraining the remainder of the disclosure in any way whatsoever. While the embodiments have been shown and described, many variations and modifications thereof can be made by one skilled in the art without departing from the spirit and teachings of the invention. Accordingly, the scope of protection is not limited by the description set out above, but is only limited by the claims, including all equivalents of the subject matter of the claims. The disclosures of all patents, patent applications and publications cited herein are hereby incorporated herein by reference, to the extent that they provide procedural or other details consistent with and supplementary to those set forth herein.

Claims

WHAT IS CLAIMED IS:

1. A method of treating or preventing a metabolic disorder in a subject, said method comprising: administering to the subject a composition comprising a therapeutically effective amount of a compound selected from the group consisting of indacaterol, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof.

2. The method of claim 1, wherein the compound comprises indacaterol, and wherein the indacaterol comprises: pharmaceutically acceptable salt thereof, a pharmaceutically acceptable hydrate thereof, or combinations thereof.

3. The method of claim 2, wherein the indacaterol comprises the following structure:

4. The method of claim 2, wherein the indacaterol is administered at a concentration of less than about 100 mg per kg of body weight.

5. The method of claim 2, wherein the indacaterol is administered at a concentration of less than about 50 mg per kg of body weight.

6. The method of claim 2, wherein the indacaterol is administered at a concentration of less than about 25 mg per kg of body weight.

7. The method of claim 2, wherein the indacaterol is administered at a concentration of less than about 15 mg per kg of body weight.

8. The method of claim 2, wherein the indacaterol is administered at a concentration of about 10 mg per kg of body weight.

9. The method of claim 1, wherein the indacaterol is co-administered with at least one glucagon- like peptide- 1 (GLP-1) agonist.

10. The method of claim 9, wherein the GLP-1 agonist is in the same composition as the indacaterol.

11. The method of claim 9, wherein the GLP-1 agonist is in a composition separate from the indacaterol.

12. The method of claim 9, wherein the GLP-1 agonist is selected from the group consisting of semaglutide, exenatide, liraglutide, albiglutide, dulaglutide, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof.

13. The method of claim 9, wherein the GLP-1 agonist comprises semaglutide.

14. The method of claim 9, wherein the GLP-1 agonist is administered at a concentration of at least about 10 nmol per kg of body weight.

15. The method of claim 9, wherein the GLP-1 agonist is administered at a concentration of at least about 50 nmol per kg of body weight.

16. The method of claim 9, wherein the GLP-1 agonist is administered at a concentration of at least about 100 nmol per kg of body weight.

17. The method of claim 1, wherein the administration occurs by a method selected from the group consisting of oral administration, inhalation, subcutaneous administration, intravenous administration, intraperitoneal administration, intramuscular administration, intrathecal injection, intra- articular administration, topical administration, central administration, peripheral administration, aerosol-based administration, nasal administration, transmucosal administration, transdermal administration, parenteral administration, intravenous administration, or combinations thereof.

18. The method of claim 1, wherein the administration comprises subcutaneous administration.

19. The method of claim 1, wherein the metabolic disorder to be treated or prevented is selected from the group consisting of obesity, diet-induced obesity, type 2 diabetes, diet-induced hyperglycemia, cardiovascular disease, hyperlipidemia, hepatic lipidosis, liver fibrosis, metabolic syndrome, insulin resistance, or combinations thereof.

20. The method of claim 1, wherein the metabolic disorder to be treated or prevented comprises obesity.

21. The method of claim 1, wherein the subject is a human being.

22. The method of claim 1, wherein the subject is obese, and wherein the obese subject has a body mass index of more than 30 (BMI >30).

23. The method of claim 1, wherein the subject is diagnosed with type 2 diabetes.

24. The method of claim 1, wherein the administration of the composition results in weight loss, restoration of glucose homeostasis, improvement in insulin sensitivity, reduced total serum cholesterol, reduced free fatty acid levels, reduced fatty liver, reduced food intake, enhanced metabolic rate, or combinations thereof.

25. A composition for use in treating or preventing a metabolic disorder in a subject, said composition comprising a therapeutically effective amount of a compound selected from the group consisting of indacaterol, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof.

26. The composition of claim 25, wherein the compound comprises indacaterol, and wherein the indacaterol comprises: , a pharmaceutically acceptable salt thereof, a pharmaceutically acceptable hydrate thereof, or combinations thereof.

27. The composition of claim 26, wherein the indacaterol comprises the following structure:

28. The composition of claim 25, wherein the compound further comprises at least one glucagon-like peptide- 1 (GLP-1) agonist.

29. The composition of claim 28, wherein the GLP-1 agonist is in the same composition as the indacaterol.

30. The composition of claim 28, wherein the GLP-1 agonist is in a composition separate from the indacaterol.

31. The composition of claim 28, wherein the GLP-1 agonist is selected from the group consisting of semaglutide, exenatide, liraglutide, albiglutide, dulaglutide, pharmaceutically acceptable salts thereof, pharmaceutically acceptable hydrates thereof, or combinations thereof.

32. The composition of claim 28, wherein the GLP-1 agonist comprises semaglutide.