Methods for fat reduction

Abstract

The present disclosure provides a method for inhibiting adipocyte differentiation and / or inducing adipocyte death in a subject, comprising administrating an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acid) to a subject in need thereof.

Description

PATENT Attorney Docket No.: G4590-18500PCT METHODS FOR FAT REDUCTION Field of the Invention

[0001] The present disclosure relates to a field of treatment. Particularly, the present disclosure provides a method for reducing body fat and / or treating and / or preventing obesity. Background of the Invention

[0002] Obesity is a burgeoning global health problem. People who are overweight or obese face increased morbidity and mortality due to serious chronic diseases including diabetes, cardiovascular diseases, stroke, dementia, metabolic syndrome and cancer. More recently, overweight and obesity have been implicated as a significant risk factors for SARS-CoV2 infection. Pathologically, obesity involves an increase in adipose tissue mass, especially white adipose, via the increase in size and / or numbers of adipocytes, the latter achieved through cellular differentiation of precursor cells into mature adipocytes (adipogenesis) or hyperplasia. Given the central role of adipocytes in adipose tissue expansion, small molecule modulators of adipocyte physiology will be invaluable both as research tools and as therapeutic agents to address obesity-linked adipocyte dysfunction and obesity-related health issues. Summary of the Invention

[0003] The present disclosure provides a method for inhibiting adipocyte differentiation and / or inducing adipocyte death in a subject, comprising administrating an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) to a subject in need thereof. The present disclosure also provides a composition comprising an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) for use in a method for inhibiting adipocyte differentiation and / or inducing adipocyte death in a subject. The present disclosure also provides a use of an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) in the G4590-18500PCT_20240918_Specification_TBF - 1 -PATENT Attorney Docket No.: G4590-18500PCT manufacture of a medicament for inhibiting adipocyte differentiation and / or inducing adipocyte death in a subject.

[0004] The present disclosure also provides a method for inducing fat tissue reduction and / or inducing cell death of adipocytes in a subject, comprising administrating an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acid) to a subject in need thereof. The present disclosure also provides a composition comprising an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) for use in a method for inducing fat tissue reduction and / or inducing cell death of adipocytes in a subject. The present disclosure also provides a use of an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) in the manufacture of a medicament for inducing fat tissue reduction and / or inducing cell death of adipocytes in a subject.

[0005] In one embodiment, the induction of fat tissue reduction does not induce inflammation-associated side effects.

[0006] In one embodiment, the nitro-fatty acids selectively induced adipocyte death.

[0007] The present disclosure also provides a method for treating and / or preventing inflammation, overweight, obesity and metabolic diseases associated with obesity and / or for governing chronic weight management, comprising administrating an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acid) to a subject in need thereof. The present disclosure also provides a composition comprising an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) for use in a method for treating and / or preventing inflammation, overweight, obesity and metabolic diseases associated with obesity and / or for governing chronic weight management. The present disclosure also provides a use of an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) in the G4590-18500PCT_20240918_Specification_TBF - 2 -PATENT Attorney Docket No.: G4590-18500PCT manufacture of a medicament for treating and / or preventing inflammation, overweight, obesity and metabolic diseases associated with obesity and / or for governing chronic weight management.

[0008] In one embodiment, the treatment and / or prevention of overweight, obesity and metabolic diseases associated with obesity and / or governance of chronic weight management does not induce inflammation-associated side effects.

[0009] In some embodiments, the metabolic diseases associated with obesity include, but are not limited to, ectopic fat accumulation, lipomas, insulin resistance, gastroesophageal reflux disease, cellulite, proteus syndrome, obesity caused by phosphatase and tensin homolog (PTEN) mutation(s), Dercum's disease, Gardner syndrome, submental fat, cardiometabolic disease, hyperuricemia, hepatic steatosis, nonalcoholic steatohepatitis, cardiovascular disease, prediabetes, osteoarthritis, obstructive sleep apnea, hepatic steatosis, type 2 diabetes, cardiovascular diseases, high blood pressure, high LDL cholesterol, low HDL cholesterol, high levels of triglycerides (dyslipidermia), coronary heart disease, gallbladder disease, osteoarthritis, stroke, sleep apnea and breathing problems and cancers.

[0010] The present disclosure also provides a method for treating, preventing and / or alleviating obesity-associated liver, pancreas and kidney pathologies induced by high-fat diets, comprising administrating an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro- fatty acid) to a subject in need thereof. The present disclosure also provides a composition comprising an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) for use in a method for treating, preventing and / or alleviating obesity-associated liver, pancreas and kidney pathologies induced by high-fat diets. The present disclosure also provides a use of an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) in the G4590-18500PCT_20240918_Specification_TBF - 3 -PATENT Attorney Docket No.: G4590-18500PCT manufacture of a medicament for treating, preventing and / or alleviating obesity-associated liver, pancreas and kidney pathologies induced by high-fat diets.

[0011] In one embodiment, the treatment, prevention and / or alleviation of obesity-associated liver, pancreas and kidney pathologies induced by high-fat diets does not induce inflammation- associated side effects.

[0012] In some embodiments, examples of the nitro-fatty acid described herein include, but are not limited to, nitro-oleic acid, (NO2-OA), nitro-palmitoleic acid (NO2-POA), nitro linolenic acid (NO2-LNA), nitro-arachidonic acid (NO2-AA), nitro eicosapentaenoic acid (NO2-EPA), nitro-docosahexaenoic acid (NO2-Dha), nitro-hydroxy acid, and nitro-allyl fatty acid and any combination thereof.

[0013] In some further embodiments, the nitro-fatty acid described herein is 9-nitro-oleic acid (9N-OA), 10-nitro-oleic acid (10N-OA), 12-nitro-oleic acid (12N-OA), 9-nitro-9-cis- octadecenoic acid, 10-nitro-9-cis-octadecenoic acid, 9-nitro-9,12-octadecadienoic acid, 10-nitro- 9,12-octadecadienoic acid, 12-nitro-9,12- octadecadienoic acid, 13-nitro-9,12-octadecadienoic acid, 9-nitrolinoleic acid, 10-nitrolinoleic acid, 12-nitrolinoleic acid, 13-nitrolinoleic acid, 5- nitro-5,8,11,14-eicosatetraenoic acid, 6-nitro-5,8,11,14-eicosatetraenoic acid, 8-nitro-5,8,11,14- eicosatetraenoic acid, 9-nitro-5,8,11,14-eicosatetraenoic acid, 11-nitro-5,8,11,14-eicosatetraenoic acid, 12-nitro-5,8,11,14-eicosatetraenoic acid, 14-nitro-5,8,11,14-eicosatetraenoic acid, 15-nitro- 5,8,11,14-eicosatetraenoic acid, 10-nitro-4,7,10,13,16,19-docosahexaenoic acid, 11-nitro- 4,7,10,13,16,19-docosahexaenoic acid, 13-nitro-4,7,10,13,16,19-docosahexaenoic acid, 14-nitro- 4,7,10,13,16,19-docosahexaenoic acid, 16-nitro-4,7,10,13,16,19-docosahexaenoic acid, 17-nitro- 4,7,10,13,16,19-docosahexaenoic acid, 19-nitro-4,7,10,13,16,19-docosahexaenoic acid or 20- nitro-4,7,10,13,16,19-docosahexaenoic acid and any combination thereof. G4590-18500PCT_20240918_Specification_TBF - 4 -PATENT Attorney Docket No.: G4590-18500PCT

[0014] In a further embodiment, the nitro-fatty acid described herein is 9-nitro-oleic acid (9N-OA) or 10-nitro-oleic acid (10N-OA) or a combination thereof.

[0015] In some embodiments, the electrophilic (unsaturated) fatty acids (such as electrophilic nitro-fatty acids) can be administered separately, concurrently or subsequently with a second agent selected from anti-cholesterol drug, antidiabetic drug and anti-obesity drug.

[0016] In some embodiments, the electrophilic fatty acids described herein include, but are not limited to, nitro-fatty acids (NO2-FA) and oxo-fatty acids. US 9750725B2 is incorporated as reference. In some embodiments, the electrophilis fatty acid has the following formula:R1is a heterocyclyl; is an optional double bond; W is —OH, —C(O)H, —C(O), —C(O)Rp, —COOH, —COORp, —Cl, —Br, —I, —CF3, — CN, —SO3, —SO2Rp, —SO3H, —NH3+, —NH2Rp+, —NRpRqRt, —NO2, ═O, ═NRp, ═CF2, or —CHF; each V is, independently —CH or —C—, wherein: V is —CH— when W is —OH, —H, —C(O)H, —C(O), —C(O)Rp, —COOH, —COORp, — Cl, —Br, —I, —F, —CF3,—CN, —SO3,—SO2Rp, —SO3H, —NH3+, —NH2Rp+, —NRpRqRtor —NO2;or V is —C— when W is ═O, ═NRp, ═CF2or ═CHF; Rpand Rqare each, independently, H, (C1-C8) alkyl, aryl, or (C1-C8) haloalkyl; Rtis (C1-C8) alkyl, aryl, or (C1-C8) haloalkyl; G4590-18500PCT_20240918_Specification_TBF - 5 -PATENT Attorney Docket No.: G4590-18500PCT Rband Rb′are each independently, —H, —OH, —C(O)H, —C(O), —C(O)Rp, —COOH, — COORp, —Cl, —Br, —I, —F, —CF3,—CHF2,—CH2F, —CN, —SO3,—SO2Rp, SO3H, — NH3+, —NH2Rp+, —NRpRqRtor —NO2, wherein: Rband Rb′are not simultaneously non-hydrogen groups; a is an integer between 5 and 15 inclusive, c is an integer from 1 and 15 inclusive, and f is an integer between 5 and 15 inclusive.

[0017] In some embodiments, W is —H, ═O or ═NRp; V is —CH— and W is —H; V is — C— and W is ═O; or c is 1, V is —C—, and W is ═O.

[0018] In some embodiments, Rband Rb′, are each, independently, —H, —OH, —CN, or — NO2;or Rband Rb′are each —H.

[0019] In some embodiments, a is 3.

[0020] In some embodiments, R1is a heteroaryl or a heterocycloalkyl.

[0021] In some further embodiments, the electrophilic fatty acid iucludes but is not limited to 13-oxo-(7Z,10Z,14A,16Z,19Z)-docosa-7,10,14,16,19-pentanoic acid, 17-oxo- (7Z,10Z,13Z,15A,19Z)-docosa-7,10,13,15,19-pentanoic acid, 13-oxo- (4Z,7Z,10Z,14A,16Z,19Z)-docosa-4,7,10,14,16,19-hexanoic acid and 17-oxo- (4Z,7Z,10Z,13Z,15A,19Z)-docosa-4,7,10,13,15,19-hexanoic acid, where A indicates either E or Z configuration, and any combination thereof. Brief Description of the Drawings

[0022] Figures 1 (A) to (C) show that nitro-oleate / fatty acids selectively kill hMSCs-derived adipocytes. (A) Proportion of live MSCs (left panel) or MSCs-derived adipocytes (right panel) after 48 h treatment with different concentrations of the indicated lipids. Error bars, S.E.M. *p<0.05, **p<0.01. n=3. (B) Cellular imaging of total (Hoechst) and dead (propidium iodide, PI) G4590-18500PCT_20240918_Specification_TBF - 6 -PATENT Attorney Docket No.: G4590-18500PCT adipocytes upon treatment with 5 μM of the different lipids or 20 μM of DCA for the indicated amount of time. Scale bar, 50 μm. n=3. (C) Proportion of dead cells at different times after lipid treatment calculated from the cellular images obtained from B. Error bars, S.E.M. *p<0.05, **p<0.01, n=3.

[0023] Figures 2 (A) and (B) show viability assay of (A) DCA-treated hMSCs and MDAs (B) FA and N-OA-treated hMSCs-derived osteoblasts.

[0024] Figures 3 (A) to (D) show that nitro-oleate-mediated adipocyte killing does not induce inflammation. (A) Production of pro-inflammatory cytokines (IL-6, IL-1β, and TNFα) by hMSCs-derived adipocytes after 48 h treatment with 5 μM of indicated lipids or 20 μM of DCA. Error bars, S.E.M. *p<0.05, **p<0.01, n=3. (B) Proliferation of PHA-treated T cells alone with indicated lipids or (C) after co-culturing with lipid-treated hMSCs-derived adipocytes. (D) IL-2 and IFNγ production by PHA-activated T cells after co-culturing with lipid-treated hMSCs- derived adipocytes. Error bars, S.E.M. *p<0.05, **p<0.01. n=3.

[0025] Figures 4 (A) to (D) show weight loss and fat reduction effect of N-OA in HFD mice. (A) Schematic graph for the generation of HFD mice (left panel) and the plan for lipid treatment (right panel). C: Carbohydrate, F: Fat, P: Protein. (B) Body weight change over the course of lipid administration (2 weeks), which is expressed in the actual body weight (left panel) or the percentage change (right panel) of body weight compared to the start of the drug treatment, n=4. ***P<0.001 (One-way analysis of variance, followed by Dunnett’s post-hoc test) (C) Representative photographs of vehicle or N-OA-treated mice fed with the indicated diets after sacrifice. The abdominal fat pads (epididymal fat) were dissected, weighed and normalized to individual body weight (% body weight), n=4. ns, not significant, *p < 0.05, ***p < 0.001 (two- sided unpaired t-test) (D) Representative H&E images of tissue from the abdominal fat pads (top G4590-18500PCT_20240918_Specification_TBF - 7 -PATENT Attorney Docket No.: G4590-18500PCT panel) and the measured sizes of adipocytes (bottom panel) from vehicle or N-OA-treated mice fed with the indicated diets. Scale bar, 50 μm, n=4. ***p < 0.001 (two-sided unpaired t-test).

[0026] Figures 5 (A) to (E) show that intraperitoneally administered N-OA ameliorates HFD-induced liver, pancreas, and kidney abnormalities. (A) Biochemical analyses of total cholesterol, triglyceride, GOT (AST), and GPT (ALT) levels in the plasma of vehicle or N-OA- treated mice fed with the indicated diets. n=4. *p < 0.05, **p < 0.01, ***p < 0.001 (two-sided unpaired t-test) (B) Livers from vehicle or N-OA-treated mice fed with the indicated diets. n=4. (C-E) Representative H&E images of the liver tissue (C), pancreatic tissue (D), and kidney tissue (E) from vehicle or N-OA-treated mice fed with the indicated diets. Blue asterisks, HFD- induced abnormality in pancreas islet. Black arrow, HFD-induced abnormality in the kidney glomerulus. Red arrow, HFD-induced abnormality abnormality in the renal tubule. Scale bars in (C), 50 μm. Scale bars in (D) and (E), 25 μm.

[0027] Figures 6 (A) to (F) show N-OA induced fat reduction of local fat tissue through intra-fat injection. (A) Schematic graph for the lipid treatment through intra-fat injection in HFD- fed mice. (B) Percentage change of body weight compared to the Day 0 over the 2-week lipid treatment. n=3. ***P<0.001 (One-way analysis of variance, followed by Dunnett’s post-hoc test) (C) Weekly food intake per mouse during lipid treatment (2 weeks). n=3, ns, not significant versus vehicle (two-sided unpaired t-test) (D) Representative photos of the abdomen areas of vehicle, N-OA, or DCA-treated HFD mice after sacrifice (above). (E) Representative photos of dissected abdominal fat pads (top panel) and weight of the fat pads (bottom panel) from vehicle, NOA, or DCA-treated HFD mice. n=3, *P<0.05 (two-sided unpaired t-test). (F) Representative H&E images of abdominal fat pads. The black arrow points to crown-like structures indicative of dead adipocytes. Scale bars, 25 μm. G4590-18500PCT_20240918_Specification_TBF - 8 -PATENT Attorney Docket No.: G4590-18500PCT

[0028] Figures 7 (A) to (F) show intra-fat administration of N-OA harbors systematical effects in reducing fat accumulation in the liver, pancreas, and kidney. (A, B) Biochemical analyses of total cholesterol, triglyceride, GOT (AST), and GPT (ALT) levels in the plasma of mice with the indicated treatments. n=3, ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001 (two-sided unpaired t-test) (C) Representative livers from the mice after sacrifice. (D-F) The liver (D), pancreas (E), and kidney (F) from mice treated with the indicated lipids were sectioned and stained with hematoxylin & eosin (top panels) or with a combination of DPAI (DNA, blue) and BODIPY (lipid droplet, green) (bottom panels). Black scale bars, 50 μm. White scale bars, 25 μm. Detailed Description of the Invention

[0029] The present disclosure shows that in addition to inhibiting adipocyte differentiation, nitro-fatty acids selectively induced the cell death of human mesenchymal stem cells (hMSCs)- derived adipocytes. The present disclosure demonstrates that nitro-oleate (N-OA) directly reduced fat tissue by triggering adipocyte death, while exerting beneficial systemic effects in alleviating obesity-associated liver, pancreas and kidney pathologies induced by high-fat diets. Notably, the present disclosure shows that the adipocyte death and fat tissue reduction induced by N-OA does not induce obvious inflammation-associated side effects normally observed for deoxycholic acid (DCA). These results highlight that N-OA is an improved therapeutic agent for fat-reduction compared to FDA-approved drugs (such as DCA). Overall, the present disclosure provides insights into the biological roles and applications of nitro-fatty acids as chemical modulators of the adipose tissue and obesity-related metabolic disorders.

[0030] Small molecule modulators of adipocyte differentiation or viability are viewed as a way to manage obesity and obesity-related diseases. Excluding extracts, a number of natural G4590-18500PCT_20240918_Specification_TBF - 9 -PATENT Attorney Docket No.: G4590-18500PCT and synthetic compounds are shown to inhibit adipogenic differentiation and lipogenesis, and / or induce apoptosis in terminally differentiated mature adipocytes. Some of these compounds require particularly high (>100 µM) concentrations to achieve the reported effects in vitro, usually in murine 3T3-L1 preadipocytes and adipocytes. Even fewer compounds have been shown to work on primary or human adipocytes or in vivo. Approved by the FDA for submental fat removal by inducing adipocyte lysis, deoxycholic acid (DCA) remains the benchmark for fat- reducing compounds despite the high doses required and common inflammation-associated side effects in the clinic.

[0031] Produced in cells as a result of inflammation, digestion or cellular metabolism, electrophilic nitro-fatty acids confer beneficial anti-inflammatory and cytoprotective effects in disease models characterized by dysfunctional metabolism. While nitro-fatty acids have been shown to accumulate and be metabolized in adipose tissues in vivo, the effects of nitro-fatty acids on the physiology of adipocytes and adipose tissue, which are central to our systemic metabolism, remains unclear and to be determined.

[0032] The present disclosure demonstrates the selective killing effects of nitro-fatty acids or electrophilic fatty acids on adipocytes in vitro and in vivo. In addition to inhibiting adipocyte differentiation, the present disclosure shows that nitro-fatty acids or electrophilic fatty acids selectively induced cell death of adipocytes via a different mechanism from FDA-approved fat-reducing agent DCA. The present disclosure demonstrates that nitro-oleate (N-OA) or electrophilic fatty acids directly reduced fat tissue by inducing adipocyte death, while exerting beneficial systemic effects in alleviating obesity-associated liver, pancreas and kidney pathologies induced by high-fat diets. Notably, the present disclosure shows that N-OA- or electrophilic fatty acid-induced adipocyte death and fat tissue reduction did not induce the G4590-18500PCT_20240918_Specification_TBF - 10 -PATENT Attorney Docket No.: G4590-18500PCT obvious inflammation-associated side effects normally observed for DCA, highlighting the possibility of N-OA or electrophilic fatty acids being an improved therapeutic agent for fat- reduction. The present disclosure provides a novel application of nitro-fatty acids or electrophilic fatty acids and informs the design of small molecule modulators of the adipose tissue for therapeutic intervention of obesity and related diseases.

[0033] Electrophilic fatty acid nitroalkenes (NO2-FA) are products of nitric oxide and nitrite- mediated unsaturated fatty acid nitration. These electrophilic products induce pleiotropic signaling actions that modulate metabolic and inflammatory responses in cell and animal models. Nitro fatty acids is a class of compounds that are formed by the reaction of unsaturated fatty acids (UFAs) (such as oleic (OA), linoleic (LA), etc.) with secondary products of nitrogen monoxide (NO.) and nitrite anions (NO2−). Examples of the nitro fatty acids include, but are not limited to, In some embodiments, examples of the nitro-fatty acid described herein include, but are not limited to, nitro-oleic acid, (NO2-OA), nitro-palmitoleic acid (NO2-POA), nitro linolenic acid (NO2-LNA), nitro-arachidonic acid (NO2-AA), nitro eicosapentaenoic acid (NO2-EPA), nitro-docosahexaenoic acid (NO2-Dha), nitro-hydroxy acid, and nitro-allyl fatty acid and any combination thereof.

[0034] Examples of the electrophilic fatty acids include, but are not limited to, those disclosed in US 9750725B2 that is incorporated into reference. US 9750725B2 demonstrates the following compounds:G4590-18500PCT_20240918_Specification_TBF - 11 -PATENT Attorney Docket No.: G4590-18500PCT In Formula (I), X is selected from the group consisting of —CH2—, —OH, —S, —ORtand — NRpRq; Y is selected from the group consisting of —C(O)—, O, —S—, and —NRpRq; W is selected from the group consisting of —OH, —H, ═S, —SRp, —C(O)H, —C(O), —C(O)Rp, — COOH, —COORp, —Cl, —Br, —I, —F, —CF3, —CN, —SO3, —SO2Rp, —SO3H, —NH3+, — NH2Rp+, —NRpRqRt, NO2, ═O, ═NRp, ═CF2, and ═CHF and V is —CH— when W is selected from the group consisting of —OH, —H, —C(O)H, —C(O), —C(O)Rp, —COOH, —COORp, —Cl, —Br, —I, —F, —CF3, —CN, —SO3, —SO2Rp, —SO3H, —NH3+, —NH2Rp+, —NRpRqRtand NO2and V is —C— when W is selected from the group consisting of ═O, ═NRp, ═CF2, and ═CHF. The indices a, b, c, d, e, and f independently are integers between 0 and 15 inclusive. In one embodiment c is 0 when d is not 0. Alternatively, d is 0 when c is not 0; such that the sums (a+b+c+e+f) and (a+b+d+e+f) independently are equal to an integer that conforms to the formula 2n or 2n+1, wherein n is an integer between 3 and 15 inclusive. Substituents —Rp, —Rqand —Rtare independently selected from H, (C1-C8)alkyl and (C1- C8)haloalkyl. In Formula I —Ra, —Ra′, —Rb, —Rb′, —Rc, —Rc′, are independently selected from the group consisting of —H, —OH, —C(O)H, —C(O), —C(O)Rp, —COOH, —COORp, —Cl, —Br, — I, —F, —CF3, —CHF2, —CH2F, —CN, —SO3, —SO2Rp, —SO3H, —NH3+, —NH2Rp+, — NRpRqRtand NO2. Additionally, —Raand —Ra′do not simultaneously represent non-hydrogen groups; —Rband — Rbdo not simultaneously represent non-hydrogen groups; and, similarly, —Rcand —Rc′do not simultaneously represent non-hydrogen groups. G4590-18500PCT_20240918_Specification_TBF - 12 -PATENT Attorney Docket No.: G4590-18500PCT In Formula I, an optional double bond is indicated by , while , when present, together with X and Y and the carbon atom to whicha 5- to 6-membered heterocyclyl or heteroaryl ring.

[0035] Particularly, the representative compounds include, but are not limited to, 13-oxo- (7Z,10Z,14A,16Z,19Z)-docosa-7,10,14,16,19-pentanoic acid, 17-oxo-(7Z,10Z,13Z,15A,19Z)- docosa-7,10,13,15,19-pentanoic acid, 13-oxo-(4Z,7Z,10Z,14A,16Z,19Z)-docosa- 4,7,10,14,16,19-hexanoic acid, 17-oxo-(4Z,7Z,10Z,13Z,15A,19Z)-docosa-4,7,10,13,15,19- hexanoic acid, where A indicates either E or Z configuration.

[0036] The electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids), or its composition as described herein can be can be formulated with a suitable carrier in various forms, such as solids, solutions, powders, fluid emulsions, fluid suspensions, semi-solids, and dry powders containing an effective amount of the electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids). These formulations may also include pharmaceutically acceptable diluents, fillers, disintegrants, binders, lubricants, surfactants, hydrophobic vehicles, water soluble vehicles, emulsifiers, buffers, humectants, moisturizers, solubilizers, antioxidants, preservatives, and similar ingredients. The means and methods for administration are well- known in the art, and guidance can be found in various pharmacologic references.

[0037] The electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids), or its composition as described herein can be formulated for parenteral or intravenous administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection can be presented in unit dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions can take such forms as suspensions, solutions or G4590-18500PCT_20240918_Specification_TBF - 13 -PATENT Attorney Docket No.: G4590-18500PCT emulsions in oily or aqueous vehicles, and can contain formulatory agents such as suspending, stabilizing and / or dispersing agents.

[0038] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent.

[0039] The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic diluent or solvent suitable for injection, such as a solution in 1,3- butanediol. Acceptable vehicles and solvents that can be used include water, Ringer's solution, and isotonic sodium chloride solution. Additionally, sterile fixed oils are commonly used as a solvent or suspending medium. Any mild fixed oil can be used, including synthetic mono- or diglycerides. Fatty acid diluents like oleic acid are also used in injectable preparations. Other fatty acid diluents that may be useful include stearic acid, metallic stearate, sodium stearyl fumarate, fatty acid, fatty alcohol, fatty acid ester, glyceryl behenate, mineral oil, vegetable oil, paraffin, leucine, silica, silicic acid, talc, propylene glycol fatty acid ester, polyethoxylated castor oil, polyethylene glycol, polypropylene glycol, polyalkylene glycol, polyoxyethylene-glycerol fatty ester, polyoxyethylene fatty alcohol ether, polyethoxylated sterol, polyethoxylated castor oil, polyethoxylated vegetable oil, and similar substances.

[0040] The formulations are solid dosage forms for oral administration including capsules, tablets, pills, powders, and granules. In such embodiments, the electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids), or its composition as described herein may be admixed with one or more inert diluent such as sucrose, lactose, or starch. Such dosage forms may also comprise, as in normal practice, additional substances other than inert diluents, e.g., G4590-18500PCT_20240918_Specification_TBF - 14 -PATENT Attorney Docket No.: G4590-18500PCT lubricating agents such as magnesium stearate. In the case of capsules, tablets, and pills, the dosage forms may also comprise buffering agents and can additionally be prepared with enteric coatings.

[0041] Additional embodiments that could be beneficial for administering the the electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty), or its composition orally include liquid forms of dosage. In these embodiments, a liquid dosage could consist of a pharmaceutically acceptable emulsion, solution, suspension, syrup, or elixir containing inert diluents commonly used in the field, such as water. These compositions may also contain adjuvants, such as wetting agents, emulsifying and suspending agents, as well as sweetening, flavoring, and perfuming agents.

[0042] The electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids), or its composition as described herein can be administered through various conventional methods and routes where they are effective. Administration can be either systemic or local, such as parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, transdermal, oral, buccal, ocular, intravaginal, inhalation, depot injections, or implants. In some cases, the administration may be parenteral or intravenous, with or without stabilizing additives to enhance systemic uptake, tissue half-life, and intracellular delivery. Hence, the electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids), or its composition as described herein can be administered in various ways (either alone or in combination with other pharmaceuticals), such as through injection (including short-acting, depot, implant, and pellet forms administered subcutaneously or intramuscularly).

[0043] In summary, the electrophilic (unsaturated) fatty acids (such as an electrophilic nitro- fatty acids) selectively induced adipocyte death while impairing adipogenesis in vitro using G4590-18500PCT_20240918_Specification_TBF - 15 -PATENT Attorney Docket No.: G4590-18500PCT MDAs. The fat-reducing activity of N-OA in diet-induced obesity mouse models without obvious inflammation-related side effects triggered by adipocyte death. The study sets the framework to understand the novel applications of the electrophilic (unsaturated) fatty acids (such as an electrophilic nitro-fatty acids) as therapeutic agents to for fat reduction and to pharmacologically manage obesity and obesity-related diseases. Examples

[0044] Example 1 Nitro fatty acids selectively kill human adipocytes

[0045] Since hMSCs are a major source of adipocyte progenitors in vivo, we chose to use hMSCs-derived adipocytes (MDAs) to explore the roles of nitro-fatty acids on the differentiation and physiology of human adipocytes. After hMSCs differentiation in a defined adipogenic medium, we obtained lipid droplet-containing cells expressing the relevant adipocyte markers. Functionally similar to mature human fat cells, MDAs provide an excellent model system to study the effects of chemical compounds on adipogenesis and adipocyte physiology.

[0046] Treatment of hMSCs and MDAs with different lipids revealed that nitro-linoleic acid and nitro-oleate (N-OA) rapidly and selectively induced the death of MDAs at low micromolar concentrations (Table 1). The induction of cell death by nitro-fatty acids is specific to MDAs as these nitro-fatty acids did not affect the viability of MSCs (Figure 1A) and hMSC-derived osteoblasts (Figure 2B). This killing effect is independent of the peroxisome proliferator- activated receptor γ (PPARγ) agonist effect reported for nitro-fatty acids,30,31 since high- affinity PPARγ ligand rosiglitazone did not affect MDAs survival. Additionally, the electrophilic nitro moiety is required since killing of MDAs was not observed with oleate (OA) and linoleate (Figure 1A). Molecular recognition of the nitro-fatty acids is also important since MDAs viability was unaffected by another electrophilic lipid, 15-deoxy-∆-12,14-prostaglandin J2 (15d- G4590-18500PCT_20240918_Specification_TBF - 16 -PATENT Attorney Docket No.: G4590-18500PCT PGJ2). 15d-PGJ2 also covalently modifies nucleophilic amino acid residues of cellular proteins via Michael addition, including a number of proteins also targeted by nitro-fatty acids such as KEAP1 and p65 subunit of NF-κB. Last but not least, compared to DCA, a bile acid that induces non-specific cell lysis for localized fat reduction, N-OA showed significantly increased killing of MDAs within 36 h at much lower CC50 values (Figure 2A, Table 1). Table 1. CC50values of indicated compounds in the hMSCs, MDAs and osteoblasts. CompoundshMSCsMDAs Osteoblasts (CC50)(CC50) (CC50)

[0047] Example 2 Nitro-oleate killing of adipocytes does not induce a pro- inflammatory response

[0048] Given that obesity-related diseases are linked to chronic inflammation of adipose tissue, and that adipocyte death is linked to adipose tissue inflammation in obesity, we next determined if N-OA induced adipocyte death triggers a pro-inflammatory response.

[0049] We first tested for the release of proinflammatory cytokines by adipocytes treated with different lipids. For DCA, MDAs death was accompanied by significantly increased levels of IL-6, IL-1β, and TNF-α in the culture supernatants (Figure 3A). This is in line with clinical observations that local administration of DCA often causes side effects like pain and swelling at injection sites. In contrast, no increase in IL-1β, and TNF-α were observed for MDAs treated with nitro-oleic acid (Figure 3A). The modest increase in IL-6 levels observed for N-OA treated G4590-18500PCT_20240918_Specification_TBF - 17 -PATENT Attorney Docket No.: G4590-18500PCT MDAs was unrelated to adipocyte death since this increase was also seen for OA treatment, which did not kill adipocytes. These results suggest a lack of an inflammatory response to adipocyte death triggered by N-OA and mechanistic differences between N-OA and DCA in the induction of adipocyte death.

[0050] Since adipocytes can regulate T lymphocyte-mediated adaptive immunity in various metabolic diseases, we tested the effects of lipid-induced adipocyte death on T cell response. Consistent with the increased production of pro-inflammatory cytokines by dying adipocytes, DCA-treated MDAs promoted the proliferation of co-cultured T lymphocytes and stimulated the production of T cell activated cytokines, IFN-γ and IL-2 upon PHA activation (Figure 3C, D). On the other hand, this response was suppressed with N-OA treatment, as observed by the reduced expansion of activated T cells co-cultured with N-OA treated MDAs (Figure 3C, D). Our data further underscored the importance of the electrophilic nitro moiety of N-OA as N-OA could inhibit the activation of T cells with or without co-culture with adipocytes in contrast to OA, which generally enhanced the T cell response (Figure 3B, C).

[0051] Overall, these results demonstrated that unlike DCA, N-OA did not generate a highly pro-inflammatory environment upon adipocyte killing, suggesting mechanistic differences in how N-OA and DCA trigger the death of human adipocytes. These results also suggest the possibility of less inflammation-associated side effects when using nitro-fatty acids instead of DCA for the reduction of adipose tissues in vivo.

[0052] Example 3 Nitro-oleate induces weight loss and fat reduction in HFD-fed mice

[0053] Given the efficient and selective killing of MDAs by N-OA in vitro, we examined ability of N-OA to reduce fat tissues in vivo using mouse models. Male C56BL / 6 mice fed on G4590-18500PCT_20240918_Specification_TBF - 18 -PATENT Attorney Docket No.: G4590-18500PCT normal-chow diet (NCD) or high-fat diet (HFD) for 12 weeks were treated with N-OA or vehicle intraperitoneally every 2 days for 2 weeks (Figure 4A). Interestingly, N-OA induced significant weight-loss in HFD mice but not NCD mice (Figure 4B), perhaps due to the lower proportion of body fat in the latter (Figure 4C). Indeed, the size and weight of inguinal fat pads were significantly reduced in N-OA-treated HFD mice compared to vehicle-treated animals (Figure 4C). Consistent with the lack of change in body weight compared to vehicle controls, the size or the weight of inguinal fat pads remained unaffected in NCD mice treated with N-OA (Figure 4C). Nonetheless, the fat-reducing effects of N-OA can still be observed in NCD mice as histological analyses of inguinal fat tissues revealed significantly reduced adipocyte sizes NCD and HFD mice treated with N-OA (Figure 4D).

[0054] We also evaluated total cholesterol (T-Cho), triglyceride (TG), glutamic oxaloacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) levels in the plasma. As expected, T-Cho, GOT, and GPT levels were higher in HFD mice compared to NCD mice, reflecting metabolic dysregulation in HFD mice (Figure 4A). Histological analyses of liver tissue from HFD mice revealed features reminiscent of non-alcoholic fatty liver disease (NAFLD), including distinct accumulation of lipid droplets and hepatocyte ballooning (Figure 5B, C). Notably, N-OA treatment significantly ameliorated the NAFLD-like features in HFD mice, lowered GOT / GPT levels and reduced hepatic lipid accumulation compared to the vehicle group (Figure 5A-C). Notably, HFD mice were also found to develop pancreas (islet) and kidney (renal glomerulus and tubule) abnormalities, which were also attenuated by systemic N-OA treatment (Figure 5D, E). G4590-18500PCT_20240918_Specification_TBF - 19 -PATENT Attorney Docket No.: G4590-18500PCT

[0055] These data demonstrate that in addition to promoting weight loss and fat reduction in vivo, intraperitoneally administered N-OA has systemic beneficial effects in alleviating obesity- associated liver, pancreas and kidney pathologies.

[0056] Example 4 Nitro-oleate induces reduction of local fat tissue through intra- fat injection.

[0057] In order to examine the direct and systemic effects of N-OA on adipocytes in vivo, we directly injected vehicle, OA, N-OA, or DCA into one of the two inguinal fat pads in HFD mice, and vehicle into the other fat pad by intra-fat injection (Figure 6A). Compared to the stable body weight of vehicle- and OA-treated mice, mice treated with N-OA and DCA showed significantly reduced body weight during treatment by almost 10% (Figure 6B). This was not due to differences in food intake since the food intake was similar for all treatment groups (Figure 6C). Notably, compared to DCA, which induced noticeable and expected local tissue swelling and hemorrhage at injection sites, no obvious side effects were observed for N-OA treated mice (Figure 6D). Consistent with their ability to kill adipocytes in vitro, we observed significant reduction in the size and weight of inguinal fat pads injected with N-OA and DCA compared to paired vehicle-treated fat pads from the same animals (Figure 6E). The appearance of characteristic crown-like structures, a specific phenomenon involving the engulfment of dead adipocytes by macrophages, is consistent with N-OA and DCA inducing adipocyte death in vivo (Figure 6F).

[0058] In addition to the direct effects on adipocytes and adipose mass as a result of local intra-fat treatments, we also noted systemic effects of N-OA. Similar to earlier results when N- OA was administered intraperitoneally, T-Cho, TG, GOT, and GPT levels were significantly reduced in animals receiving intra-fat injections of N-OA (Figure 7A). This effect specific to N- G4590-18500PCT_20240918_Specification_TBF - 20 -PATENT Attorney Docket No.: G4590-18500PCT OA since T-Cho, GOT, and GPT levels were unaffected in DCA-treated mice. Further demonstrating the importance of the electrophilic nitro-moiety on N-OA, OA treatment instead exacerbated the pathophysiological effects of HFD by elevating plasma TG and GOT levels (Figure 7A). Histological analyses of various tissues were consistent with the sera biochemical analyses. Lipid accumulation in the liver (Figure 7C, D), pancreas (Figure 8E) and kidney (Figure 7F) were strikingly reduced only in N-OA treated mice but not in OA- and DCA-treated animals.

[0059] Overall, these results indicated that N-OA directly reduced fat tissue by inducing adipocyte death in vivo, and demonstrated the systemic beneficial effects of N-OA in alleviating obesity-associated liver, pancreas and kidney pathologies. The lack of obvious inflammation- associated side effects observed for N-OA treated mice compared to DCA-treated mice was consistent with our in vitro data, and highlighted the possibility of N-OA being an improved therapeutic agent for fat-reduction.

[0060] Our animal studies showcased that using N-OA in vivo as an anti-obesity medication for weight loss and fat reduction. Intraperitoneally administrated N-OA showed significant weight loss in HFD mice while the weight of N-OA treated NCD remains unchanged. The inguinal fat tissue as well as adipocytes size were smaller HFD than in NCD mice treated with N-OA. These results uncovered that the N-OA harbored fat-targeting (or obese-targeting) capability in vivo, which is consistent with our in vitro results. N-OA also has systemic anti- obesity effects in HFD mice, including reduced blood cholesterol, HFD-induced fatty liver, and HFD-induced abnormalities in the pancreas and kidney. Overall, we found that the therapeutic potential of N-OA in reversing HFD- or obesity-induced pathogenesis in vivo. G4590-18500PCT_20240918_Specification_TBF - 21 -PATENT Attorney Docket No.: G4590-18500PCT

[0061] Topical delivery of anti-fat agents for local fat reduction is increasingly popular for tissue repair and aesthetic medicine mainly due to faster treatment outcomes and ease of handling. For injectable DCA (Kybella®), however, fat reduction is commonly accompanied by adverse side effects such as pain, swelling, warmth, numbness, and bruising at injection sites. These side effects might be due to the secretion of pro-inflammatory cytokines by the damaged adipose tissue and the activation / expansion of stimulated T lymphocytes. In contrast, proinflammatory responses were not observed with N-OA mediated adipocyte killing in vitro and in vivo, suggesting that N-OA may be a superior alternative to DCA for fat reduction in the clinic.

[0062] Direct injection of N-OA into the inguinal fat pads of HFD mice resulted in strikingly smaller fat pads without noticeable side effects. The significantly more pronounced fat loss in N- OA-treated fat pads than vehicle-treated contralateral fat pads supported the direct effects of N- OA on fat reduction. Additionally, systemic effects of N-OA were also observed. Systemic effects such as downregulation of blood cholesterol, GOT, and GPT, and reduced HFD- associated fat accumulation in multiple organs were observed in HFD mice receiving intra-fat injections of N-OA. These beneficial systemic effects are unique to N-OA as these effects were not seen in animals with pronounced fat loss as a result of intra-fat DCA injections.

[0063] Overall, our results suggested that administration of N-OA for body weight control can help alleviate metabolic disorders and diseases linked to high-cholesterol, including cardiovascular disease, diabetes, and high blood pressure, as well as fatty liver, fatty pancreas, and fatty kidney. In addition, N-OA could also be used topically to locally reduce adipose tissue with potentially fewer adverse effects than the currently used agent (DCA) in the clinic. While G4590-18500PCT_20240918_Specification_TBF - 22 -PATENT Attorney Docket No.: G4590-18500PCT oral administration of N-OA is possible and being investigated in the clinic, the effects of oral administration of N-OA on fat reduction remains to be determined.

[0064] In conclusion, we showed that nitro-fatty acids selectively induced adipocyte death while impairing adipogenesis in vitro using MDAs. We further demonstrated fat-reducing activity of N-OA in diet-induced obesity mouse models without obvious inflammation-related side effects triggered by adipocyte death, unlike FDA-approved fat reducing agent DCA. The study sets the framework to understand the novel applications of nitro-fatty acids as therapeutic agents to for fat reduction and to pharmacologically manage obesity and obesity-related diseases. G4590-18500PCT_20240918_Specification_TBF - 23 -

Claims

PATENT Attorney Docket No.: G4590-18500PCT Claims What is claimed is:

1. A composition comprising an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) for use in a method for inhibiting adipocyte differentiation and / or inducing adipocyte death in a subject.

2. A composition comprising an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) for use in a method for inducing fat tissue reduction in a subject, comprising administrating an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro- fatty) to a subject in need thereof.

3. A composition comprising an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) for use in a method for treating and / or preventing obesity and metabolic diseases associated with obesity, comprising administrating an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty) to a subject in need thereof.

4. The composition of claim 3, wherein the metabolic diseases associated with obesity include, but are not limited to, ectopic fat accumulation, lipomas, insulin resistance, gastroesophageal reflux disease, cellulite, proteus syndrome, obesity caused by phosphatase and tensin homolog (PTEN) mutation(s), Dercum's disease, Gardner syndrome, submental fat, cardiometabolic disease, hyperuricemia, hepatic steatosis, nonalcoholic steatohepatitis, prediabetes, obstructive sleep apnea, hepatic steatosis, type 2 diabetes, cardiovascular diseases, high blood pressure, high LDL cholesterol, low HDL cholesterol, high levels of triglycerides (dyslipidermia), coronary heart disease, gallbladder disease, osteoarthritis, stroke, sleep apnea and breathing problems and cancers. G4590-18500PCT_20240918_Specification_TBF - 24 -PATENT Attorney Docket No.: G4590-18500PCT 5. A composition comprising an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty acids) for use in a method for treating and / or preventing alleviating obesity-associated liver, pancreas and kidney pathologies, and the inflammatory immunity which induced by high-fat diets, comprising administrating an electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty) to a subject in need thereof.

6. The composition of any proceeding claims, wherein the nitro-fatty acid is nitro-oleic acid, (NO2-OA), nitro-palmitoleic acid (NO2-POA), nitro linolenic acid (NO2-LNA), nitro- arachidonic acid (NO2-AA), nitro-eicosapentaenoic acid (NO2-EPA), nitro-docosahexaenoic acid (NO2-Dha), nitro-hydroxy acid or nitro-allyl fatty acid or any combination thereof.

7. The composition of any proceeding claims, wherein the nitro-fatty acid is 9-nitro-oleic acid (9N-OA), 10-nitro-oleic acid (10N-OA), 12-nitro-oleic acid (12N-OA), 9-nitro-9-cis- octadecenoic acid, 10-nitro-9-cis-octadecenoic acid, 9-nitro-9,12-octadecadienoic acid, 10-nitro- 9,12- octadecadienoic acid, 12-nitro-9,12- octadecadienoic acid, 13-nitro-9,12-octadecadienoic acid, 9-nitrolinoleic acid, 10-nitrolinoleic acid, 12-nitrolinoleic acid, 13-nitrolinoleic acid, 5- nitro-5,8,11,14-eicosatetraenoic acid, 6-nitro-5,8,11,14-eicosatetraenoic acid, 8-nitro-5,8,11,14- eicosatetraenoic acid, 9-nitro-5,8,11,14-eicosatetraenoic acid, 11-nitro-5,8,11,14-eicosatetraenoic acid, 12-nitro-5,8,11,14-eicosatetraenoic acid, 14-nitro-5,8,11,14-eicosatetraenoic acid, 15-nitro- 5,8,11,14-eicosatetraenoic acid, 10-nitro-4,7,10,13,16,19-docosahexaenoic acid, 11-nitro- 4,7,10,13,16,19-docosahexaenoic acid, 13-nitro-4,7,10,13,16,19-docosahexaenoic acid, 14-nitro- 4,7,10,13,16,19-docosahexaenoic acid, 16-nitro-4,7,10,13,16,19-docosahexaenoic acid, 17-nitro- 4,7,10,13,16,19-docosahexaenoic acid, 19-nitro-4,7,10,13,16,19-docosahexaenoic acid or 20- nitro-4,7,10,13,16,19-docosahexaenoic acid or any combination thereof. G4590-18500PCT_20240918_Specification_TBF - 25 -PATENT Attorney Docket No.: G4590-18500PCT 8. The composition of any proceeding claims, wherein the nitro-fatty acid is 9-nitro-oleic acid (9N-OA) or 10-nitro-oleic acid (10N-OA) or a combination thereof.

9. The composition of any proceeding claims, wherein the nitro-fatty acid or an electrophilic fatty acid can be administered separately, concurrently or subsequently with a second agent selected from anti-cholesterol drug, antidiabetic drug and anti-obesity drug.

10. The composition of any proceeding claims, wherein the electrophilic fatty acid has the following formula,R1is a heterocyclyl; is an optional double bond; W is —OH, —C(O)H, —C(O), —C(O)Rp, —COOH, —COORp, —Cl, —Br, —I, —CF3, — CN, —SO3, —SO2Rp, —SO3H, —NH3+, —NH2Rp+, —NRpRqRt, —NO2, ═O, ═NRp, ═CF2, or —CHF; each V is, independently —CH or —C—, wherein: V is —CH— when W is —OH, —H, —C(O)H, —C(O), —C(O)Rp, —COOH, —COORp, — Cl, —Br, —I, —F, —CF3,—CN, —SO3,—SO2Rp, —SO3H, —NH3+, —NH2Rp+, —NRpRqRtor —NO2;or V is —C— when W is ═O, ═NRp, ═CF2or ═CHF; Rpand Rqare each, independently, H, (C1-C8) alkyl, aryl, or (C1-C8) haloalkyl; Rtis (C1-C8) alkyl, aryl, or (C1-C8) haloalkyl; G4590-18500PCT_20240918_Specification_TBF - 26 -PATENT Attorney Docket No.: G4590-18500PCT Rband Rb′are each independently, —H, —OH, —C(O)H, —C(O), —C(O)Rp, —COOH, — COORp, —Cl, —Br, —I, —F, —CF3,—CHF2,—CH2F, —CN, —SO3,—SO2Rp, SO3H, — NH3+, —NH2Rp+, —NRpRqRtor —NO2, wherein: Rband Rb′are not simultaneously non-hydrogen groups; a is an integer between 5 and 15 inclusive, c is an integer from 1 and 15 inclusive, and f is an integer between 5 and 15 inclusive.

11. The composition of claim 10, wherein W is —H, ═O or ═NRp; V is —CH— and W is —H; V is —C— and W is ═O; or c is 1, V is —C—, and W is ═O.

12. The method of claim 10 or 11, wherein Rband Rb′, are each, independently, —H, — OH, —CN, or —NO2;or Rband Rb′are each —H.

13. The composition of claim 10, 11 or 12, wherein a is 3.

14. The composition of any proceeding claims, wherein the electrophilic fatty acid is 13- oxo-(7Z,10Z,14A,16Z,19Z)-docosa-7,10,14,16,19-pentanoic acid, 17-oxo- (7Z,10Z,13Z,15A,19Z)-docosa-7,10,13,15,19-pentanoic acid, 13-oxo- (4Z,7Z,10Z,14A,16Z,19Z)-docosa-4,7,10,14,16,19-hexanoic acid or 17-oxo- (4Z,7Z,10Z,13Z,15A,19Z)-docosa-4,7,10,13,15,19-hexanoic acid, where A indicates either E or Z configuration or any combination thereof.

15. The composition of any proceeding claims, wherein the induction of fat tissue reduction does not induce inflammation-associated side effects.

16. The composition of any proceeding claims, wherein the electrophilic (unsaturated) fatty acid (such as an electrophilic nitro-fatty) selectively induced adipocyte death. G4590-18500PCT_20240918_Specification_TBF - 27 -

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