Resveratrol and quercetin-loaded nanoparticles and methods of use thereof

Abstract

The present invention provides nanoparticles or compositions thereof comprising resveratrol and / or quercetin, or derivatives, prodrugs, or pharmaceutically acceptable salts thereof, at least one lipid, at least one surfactant, and at least one vitamin E. In some embodiments, the nanoparticles or compositions thereof further comprise at least one targeting agent. In some embodiments, the nanoparticles or compositions thereof are targeted for an adipose tissue of interest. In some aspects, the present invention provides methods of activating brown adipose tissue or browning white adipose tissue or beige adipose tissue of interest. In some aspects, the present invention provides methods of reducing body fat, inducing weight loss, and / or improving glucose homeostasis and insulin sensitivity. In some aspects, the present invention provides methods of optimizing the composition of the gut microbiota. In some aspects, the present invention provides methods of treating or preventing various diseases or disorders, such as obesity.

Description

[0001] TITLE OF THE INVENTION

[0002] RESVERATROL AND QUERCETIN-LOADED NANOPARTICLES AND METHODS OF

[0003] USE THEREOF

[0004] CROSS-REFERENCE TO RELATED APPLICATIONS

[0005] This application claims priority to U.S. Provisional Patent Application No. 63 / 514,935, filed July 21, 2024, the disclosure of which is hereby incorporated by reference in its entirety.

[0006] REFERNCE TO A SEQUENCE LISTING SUBMITTED AS AN XML FILE This application hereby incorporates by reference the entire contents of the Sequence Listing File named “206339-0098-00WO_SequenceListing.xml” in XML format, with a creation date of July 17, 2024, and a size of 2,898 bytes.

[0007] BACKGROUND OF THE INVENTION

[0008] Prevalence of obesity and its related metabolic diseases is increasing in the U.S. and worldwide, and there is a critical need for developing an effective and safe therapeutic product. Pathologically expanded white adipose tissue (WAT) is strongly associated with increased multiple comorbidities, such as diabetes, cardiovascular disease, and certain types of cancer. Current lifestyle intervention (low compliance), pharmacotherapy (low efficacy), and bariatric surgery (high side effects and cost) have many disadvantages and cannot eradicate obesity in general populations. More practical and better approaches to combating obesity are urgently needed.

[0009] Many phytochemicals including resveratrol (R) and quercetin (Q) have antiobesity, anti-inflammation, and other beneficial effects. But their low levels of stability and bioavailability limit their applications.

[0010] Thus, there is a need in the art for compositions and methods for treating or preventing various diseases or disorders, such as obesity, obesity-related diseases or disorders, diabetes, fatty liver disease, cardiovascular disease, and other metabolic diseases and disorders. The present invention satisfies this unmet need. SUMMARY OF THE INVENTION

[0011] In one aspect, the present invention relates to a nanoparticle comprising: a) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, wherein resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, is present in a concentration range of about 0.1 wt% to about 90 wt%; b) quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, wherein quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, is present in a concentration range of about 0.1 wt% to about 90 wt%; c) at least one lipid, wherein the lipid is present in a concentration range of about 5 wt% to about 99 wt%; d) at least one surfactant, wherein the surfactant is present in a concentration range of about 10 wt% to about 75 wt%; and e) at least one vitamin E, wherein the vitamin E is present in a concentration range of about 5 wt% to about 85 wt%.

[0012] In some embodiments, the at least one lipid is selected from the group consisting of a fatty acid, wax, sterol, lipid-soluble vitamin, vitamin A, vitamin D, vitamin E, vitamin K, monoglyceride, diglyceride, triglyceride, phospholipid, phosphatidylcholine, L-a- phosphatidyl choline (PC), soy PC, egg PC, phosphatidic acid (PA), phosphatidylethanolamine (PE), lecithin, phosphatidylserine (PS), phosphoinositide, phosphatidylinositol (PI), phosphatidylinositol phosphate (PIP), phosphatidylinositol bisphosphate (PIP2), phosphatidylinositol trisphosphate (PIP3), phosphosphingolipid, sphingolipid, ceramide phosphorylcholine (SPH), ceramide phosphorylethanolamine (Cer-PE), ceramide phosphoryl lipid, and any combination thereof. In some embodiments, the at least one lipid is phosphatidylcholine.

[0013] In some embodiments, the at least one surfactant is selected from the group consisting of a polyethylene glycol (PEG), functionalized PEG, polyethylene glycol (15)-hydroxystearate , polyethylene glycol (15)-hydroxy stearate containing PEG, polysorbate 80, phospholipids, PEG21 -stearate, PEG40-stearate, PEGlOO-stearate, PEG(10-1000)-fatty acid, mustard, lecithin, soy lecithin, egg lecithin, monoglycerides, diglycerides, polysorbates, carrageenan, guar gum, canola oil, polysorbates, sodium dodecyl sulfate, lauryl dimethyl amine oxide, cetyltrimethylammonium bromide (CTAB), poly ethoxylated alcohols, polyoxyethylene sorbitan, octoxynol, N, N-dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide (HTAB), polyoxyl 10 lauryl ether, bile salts, sodium deoxy cholate, sodium cholate, polyoxyl castor oil, nonylphenol ethoxylate, cyclodextrins, methylbenzethonium chloride, and any combination thereof. In some embodiments, the at least one surfactant is polyethylene glycol (15)-hy dr oxy stearate , polyethylene glycol (15)-hydroxy stearate containing PEG.

[0014] In some embodiments, the at least one vitamin E is selected from the group consisting of a-tocopherol, a-tocopherol acetate (aTA), a-tocopherol nicotinate, D-a-tocopheryl polyethylene glycol 1000 succinate, P-tocopherol, P-tocopherol acetate, P-tocopherol nicotinate, y-tocopherol, y-tocopherol acetate, ' / -tocopherol nicotinate, 5-tocopherol, 6-tocopherol acetate, 8-tocopherol nicotinate, a-tocotrienol, a-tocotrienol acetate, a-tocotrienol nicotinate, P-tocotrienol, P- tocotrienol acetate, P-tocotrienol nicotinate, y-tocotrienol, y-tocotrienol acetate, y-tocotrienol nicotinate, 5-tocotrienol, 8-tocotrienol acetate, 8-tocotrienol nicotinate, and any combination thereof. In some embodiments, the at least one vitamin E is aTA.

[0015] In some embodiments, the nanoparticle further comprises at least one targeting agent. In some embodiments, the at least one targeting agent targets an adipose tissue. In some embodiments, the adipose tissue is a white adipose tissue (WAT), beige adipose tissue, brown adipose tissue (BAT), or any combination thereof. In some embodiments, the targeting agent is selected from the group consisting of proteins, peptides, antibodies, lipids, polysaccharides, nucleic acids, aptamers, and small molecules.

[0016] In some embodiments, the composition further comprises at least one phytochemical or bioactive compound selected from the group consisting of curcumin, theaflavins, thearubigins, epigallocatechin gallate (EGCG), (+)-catechin, (-)-epicatechin, (-)-epicatechin gallate, (-)- epigallocatechin, (+)-gallocatechin, isorhamnetin, kaempferol, myricetin, apigenin, luteolin, baicalein, chrysin, forskolin, chlorophyll a, chlorophyll b, eriodictyol, hesperetin, naringenin, taxifolin, catechins, luteolin, cyanidin, genistein, daidzein, genistein, glycitein, biochanin A, formononetin, cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, proanthocyanidins, a-carotene, P-carotene, -cryptoxanthin, lutein, zeaxanthin, lycopene, and any combination thereof.

[0017] In some embodiments, the nanoparticle further comprises at least one coating agent. In some embodiments, the nanoparticle is a biodegradable nanoparticle. In some embodiments, the nanoparticle increases bioavailability of resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0018] In some embodiments, the nanoparticle: induces browning of a white adipose tissue (WAT), beige adipose tissue, or any combination thereof; increases the level or activity of a brown adipose tissue (BAT), beige adipose tissue, or any combination thereof; increases the concentration of cellular cyclic adenosine monophosphate (cAMP); increases the level or activity of uncoupling protein 1 (UCP1) or PR domain containing 16 (PRDM16); increases the level or activity of a 5' adenosine monophosphate-activated protein kinase (AMPK); increases the level or activity of sirtuin 1 (SIRT1); increases the level or activity of tight junction protein-related genes; increases the level or activity of occluding (OCLN); increases the level or activity of peroxisome proliferator-activated receptor gamma coactivator 1 -alpha (PGC-la); decreases the level or activity of a fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), or a combination thereof; decreases the level or activity of NF-KB, tumor necrosis factor alpha (TNF-a), interferon-y, Interleukin 6 (IL-6), IL-4, IL- la, monocyte chemoattractant protein 1 (MCP1), or a combination thereof; decreases the level or activity of a phosphodiesterase (PDE); or any combination thereof.

[0019] In some embodiments, the nanoparticle: increases a mitochondrial biogenesis, mitochondrial function, thermogenesis, or a combination thereof; increases gut integrity, gut health, or a combination thereof; increases insulin sensitivity; increases energy expenditure of at least one cell of interest; improves metabolic health; improves gut microbiota profile; improves glucose homeostasis, lipid homeostasis, or a combination thereof; decreases fat mass and body weight; decreases inflammatory response; decrease liver damage; or any combination thereof.

[0020] In some embodiments, the nanoparticle does not change the level or concentration of lean mass.

[0021] In some embodiments, the present invention provides a composition comprising at least one nanoparticle of the present invention. In some embodiments, the composition is selected from the group consisting of an oral formulation, an injectable solution, a solution for transdermal delivery, an injectable hydrogel, a surgically implantable hydrogel, and any combination thereof.

[0022] In some embodiments, the present invention provides a method of increasing the concentration of a cellular cyclic adenosine monophosphate (cAMP) in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle of the invention; at least one composition of the invention; or resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof. In some embodiments, the method increases the concentration of a cellular cyclic adenosine monophosphate (cAMP) in an adipose tissue.

[0023] In some embodiments, the present invention provides a method of selectively delivering resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof to an adipose tissue, wherein the method comprises administering: at least one nanoparticle of the invention; at least one composition of the invention; or resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0024] In some embodiments, the present invention provides a method of inducing browning of an adipose tissue in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle of the invention; at least one composition of the invention; or resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0025] In some embodiments, the adipose tissue of a method of the present invention is a white adipose tissue (WAT), beige adipose tissue, brown adipose tissue (BAT), or any combination thereof.

[0026] In some embodiments, the present invention provides a method of reducing body fat in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle of the invention; at least one composition of the invention; or resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0027] In some embodiments, the present invention provides a method of inducing weight loss in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle of the invention; at least one composition of the invention; or resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0028] In some embodiments, the present invention provides a method of treating or preventing a metabolic disease or disorder in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle of the invention; at least one composition of the invention; or resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof. In some embodiments, the metabolic disease or disorder is selected from the group consisting of obesity, obesity-related disease or disorder, diabetes, fatty liver disease, non-alcoholic fatty liver disease, cardiovascular disease, heart disease, stroke, hypertension, idiopathic intracranial hypertension, coronary artery disease, atrial fibrillation, atherosclerosis, venous thromboembolism, obstructive sleep apnea , obesity hypoventilation syndrome, metabolic syndrome, insulin resistance, esophageal adenocarcinoma, gastric cancer, colorectal cancer, hepatocellular carcinoma, cholangiocarcinoma, pancreatic cancer, endometrial carcinoma, ovarian cancer, breast cancer, renal cell carcinoma, multiple myeloma, and any combination thereof.

[0029] In some embodiments, a method of the present invention comprises administration via oral administration, injection, or a combination thereof.

[0030] In some embodiments, a method of the present invention the method further comprises a fecal microbiota transplantation. In some embodiments, the fecal microbiota transplantation is a human fecal microbiota transplantation (HFT).

[0031] BRIEF DESCRIPTION OF THE DRAWINGS

[0032] The following detailed description of various embodiments of the invention are better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, there are shown in the drawings illustrative embodiments. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

[0033] Figure 1, comprising Figure 1A through Figure ID, depicts representative results demonstrating that nanoencapsulation improves reseveratrol’s (R) aqueous solubility. Figure 1A depicts an illustration of R-nanoparticle (R-NP) structure and composition. Figure IB depicts representative transmission electron microscopy (TEM) images of R-NPs. Figure 1C depicts representative images demonstrating the solubility of R-NPs. Figure ID depicts representative results demonstrating the stability of R-NPs.

[0034] Figure 2, comprising Figure 2A through Figure 2G, depicts representative results demonstrating that R-NPs decrease 50% of fat mass but do not change lean mass after IV delivery. Figure 2A depicts representative body weight of animals treated with saline, vacant nanoparticles (V-NPs), free R, and R-NPs over time. Figure 2B depicts representative fat mass after treatment and % body fat of animals treated over time. Figure 2C depicts representative lean mass after treatment and % lean body mass over time. Figure 2D depicts representative food intake of animals treated over time. Figure 2E depicts representative gonadal (g) WAT and inguinal (i) WAT mass of treated animals. Figure 2F depicts representative images of H&E staining of iWAT of treated animals; scale bar = 200 pm. Figure 2G depicts representative average adipocyte size of iWAT.. For Figure 2A through Figure 2E and Figure 2G, data are presented as means ± SEM, bars or lines without a common superscript differ (p<0.05).

[0035] Figure 3, comprising Figure 3A through Figure 3D, depicts representative results demonstrating that R-NPs deliver more R to WAT and induce iWAT browning more than free R. Figure 3A depicts representative results demonstrating R content in gWAT and iWAT. Figure 3B depicts representative results demonstrating that R content in the liver is lower in animals treated with R-NPs. Figure 3C depicts representative results demonstrating that iWAT UCP-1 mRNA levels are increased with R-NP treatment. Figure 3D depicts representative images demonstrating iWAT UCP-1 protein levels in iWAT isolated from C57BL / 6J mice; scale bar = 200 pm. For Figure 3A through Figure 3C, data are presented as mean ± SEM, n = 10, bars without a common superscript differ (p<0.05).

[0036] Figure 4, comprising Figure 4A through Figure 4E, depicts representative results demonstrating that R-NPs improve glucose homeostasis and attenuate inflammation in C57BL / 6J mice. Figure 4A depicts representative fasting blood levels of insulin. Figure 4B depicts representative fasting blood levels of glucose. Figure 4C depicts representative plasma concentrations of TNF-a. Figure 4D depicts representative plasma concentrations of IL-6. Figure 4E depicts representative plasma concentrations of MCP-1. Data are means ± SEM, n = 10, bars without a common superscript differ (p<0.05). Figure 5, comprising Figure 5A and Figure 5B, depicts representative results demonstrating nanoencapsulation improves Q’s aqueous solubility. Figure 5 A depicts a representative TEM image of Q-NPs stained by 2% of uranyl acetate. Figure 5B depicts a representative image of free Q, Q-NP, V-NP suspensions in I xPBS.

[0037] Figure 6 depicts schematic images of eight treatments.

[0038] Figure 7 depicts a representative schematic showing the structures of resveratrol and quercetin and their encapsulation into nanoparticles.

[0039] Figure 8, comprising Figure 8A through Figure 8C, depicts representative results demonstrating encapsulating R within NPs can substantially augment its aqueous solubility, chemical stability, and anti-obesity effects. Figure 8A depicts representative images of the solubility of various treatments and TEM images of R-NPs, Q-NPs, and RQ-NPs. Figure 8B depicts representative in vitro release profiles of free R, free Q, free R and Q, R-NPs, Q-NPs, and RQ-NPs. Figure 8C depicts representative anti-obesity effects of free R and R-NPs.

[0040] Figure 9 depicts a schematic representation of methods of treatment examined.

[0041] Figure 10, comprising Figure 10A through Figure 10E, depicts representative experimental results demonstrating RQ-NPs decrease of body weight, fat mass and liver inflammation, and improve glucose intolerance of obese mice. Figure 10A depicts representative body weight of mice during the 10-week treatment. Figure 10B depicts representative fat mass (%) of mice in the final week. Figure 10C depicts representative average daily food intake of mice. Figure 10D depicts representative liver inflammation related genes expression. Figure 10E depicts representative AUC of glucose tolerance test.

[0042] Figure 11, comprising Figure 11A through Figure 1 ID, depicts representative results demonstrating that RQ-NPs increase thermogenesis- and gut integrity -related gene expression. Figure 11 A depicts representative thermogenesis-related gene expression in brown adipose tissue (BAT). Figure 1 IB depicts representative thermogenesis-related gene expression in inguinal white adipose tissue (iWAT) and gonadal adipose tissue (gWAT). Figure 11C depicts representative gut function-related gene expression in duodenum and jejunum. Figure 1 ID depicts representative gut function-related gene expression in ileum and colon

[0043] Figure 12 depicts representative relative abundance of bacteria in the gut microbiome at the phylum level.

[0044] Figure 13 depicts representative experimental results demonstrating the beta- diversity of the gut microbiome. Figure 13A depicts representative beta-diversity in mice fed a low-fat diet, a high-fat diet, or a high-fat diet and treated with free RQ or RQ-NPS. Figure 13B depicts representative beta-diversity in mice fed a high-fat diet with or without a human fecal transplant (HFT). Figure 13C depicts representative beta-diversity in mice fed a high-fat diet with a human fecal transplant treated with free RQ, RQ-NP, or no treatment.

[0045] DETAILED DESCRIPTION

[0046] The present invention is based, in part, on the unexpected discovery that nanoparticles comprising resveratrol and quercetin had synergistically increased anti-obesity activities. Thus, in one aspect, the present invention provides nanoparticles or compositions thereof comprising resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof, at least one lipid, at least one surfactant, and at least one vitamin E. In some embodiments, the nanoparticles or compositions thereof further comprise at least one targeting agent. In some embodiments, the nanoparticles or compositions thereof are targeted for an adipose tissue of interest. In some embodiments, the nanoparticles or compositions thereof further comprise at least one coating agent.

[0047] In some aspects, the present invention provides methods of activating brown adipose tissue of interest. In other aspects, the present invention provides methods of browning white adipose tissue and / or beige adipose tissue of interest. In various aspects, the present invention also provides methods of increasing the concentration of a cellular cAMP.

[0048] In some aspects, the present invention provides methods of reducing body fat, inducing weight loss, and / or improving glucose homeostasis and insulin sensitivity. In some aspects, the present invention provides methods of treating or preventing a metabolic disease or disorder, such as obesity, obesity-related disease or disorder, diabetes, fatty liver disease, nonalcoholic fatty liver disease, cardiovascular disease, etc.

[0049] Definitions

[0050] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention.

[0051] As used herein, each of the following terms has the meaning associated with it in this section.

[0052] The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0053] “About” as used herein when referring to a measurable value, for example numerical values and / or ranges, such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20%, ±10%, ±5%, ±1%, or ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. For example, “about 40 [units]” may mean within ± 25% of 40 (e.g., from 30 to 50), within ± 20%, ± 15%, ± 10%, ± 9%, ± 8%, ± 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, ± 1%, less than ± 1%, or any other value or range of values therein or therebelow. Furthermore, the phrases “less than about [a value]” or “greater than about [a value]” should be understood in view of the definition of the term “about” provided herein.

[0054] As used herein, the term “nanoparticle” refers to particles having a particle size on the nanometer scale (e.g., about 1 nm - 10,000 nm). For example, the nanoparticle may have a particle size up to about 2,000 nm. In another example, the nanoparticle may have a particle size up to about 100 nm. In another example, the nanoparticle may have a particle size up to about 6 nm. As used herein, “nanoparticle” refers to a number of nanoparticles, including, but not limited to, liposomes, lipid nanoparticles, polymer nanoparticles, organic nanoparticles, inorganic nanoparticles, biocompatible nanoparticles, such as biocompatible organic nanoparticles, biocompatible inorganic nanoparticles, etc., nanoclusters, nanocapsules, core-shell nanocapsules, nanovesicles, micelles, block copolymer micelles, lamaellae shaped particles, polymersomes, dendrimers, emulsions, exosomes, self-emulsifying drug delivery systems (SEDDS), microspheres, micro- structured lipid carriers, nano-structured lipid carriers, and other nano-size particles of various other small fabrications that are known to those of skill in the art. Examples of suitable nanoparticles useful in the invention include, but are not limited to, those described in Wang et al., 2014, J. Nutri. Biochem., 25:363-376, which is incorporated herein by reference in its entirety. The shapes and compositions of nanoparticles may be guided during condensation of atoms by selectively favoring growth of particular crystal facets to produce spheres, rods, wires, discs, cages, core-shell structures and many other shapes. The definitions and understandings of the entities falling within the scope of nanocapsule are known to those of skill in the art, and such definitions are incorporated herein by reference and for the purposes of understanding the general nature of the subject matter of the present application. However, the following discussion is useful as a further understanding of some of these terms.

[0055] As used herein, the phrase “lipid nanoparticle” refers to a transfer vehicle comprising at least one lipid (e.g., cationic lipids, non-cationic lipids, polar lipids, and PEG- modified lipids) and / or at least one polymers. Examples of suitable lipids include, but are not limited to, the phospholipid compounds (e.g., phosphatidylglycerol, phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, sphingolipids, cerebrosides, and gangliosides). Examples of suitable polymers include, but are not limited to, polyacrylates, polyalkycyanoacrylates, polylactide, polylactide-polyglycolide copolymers, polycaprolactones, dextran, albumin, gelatin, alginate, collagen, chitosan, cyclodextrins, dendrimers, and polyethylenimine.

[0056] The term “liposome” as used herein refers to microscopic vesicles or particles made up of at least one lipid bilayer enclosing an internal aqueous medium. To form liposomes, the presence of at least one “vesicle-forming lipid” is needed, which is an amphipathic lipid capable of either forming or being incorporated into a lipid bilayer. Any suitable vesicle-forming lipid may be used to form the lipid bilayer constituting the liposomes. Vesicle-forming lipid includes, but not limited to, phospholipids such as phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidic acid (PA), phosphatidylethanolamine (PE) or phosphatidyl serine (PS), and charged lipids, such as a positively charge lipid or a negatively charged lipid. The term “liposome” may have the same meaning and may be interchanged with the term “lipid vesicle.” Liposome examples include large unilamellar vesicles, multilamellar vesicles, paucilamellar vesicles, small unilamellar vesicles, reverse phase evaporation vesicles, French press vesicles, and ether injection vesicles. Products incorporating liposomes include, but are not limited to, adjuvants, drug carriers, and cleansers. The following references disclose non-limiting examples of methods and / or nonlimiting examples of apparatuses for manufacturing liposomes: “LIPOSOMES — Potential for Commercial Application”, by Dr. Norman D. Weiner, presented at the Emulsion-Suspension Technology Conference, Oct. 20-23, 1997, at New Brunswick, N.J.; U.S. Pat. No. 4,911,928 to Wallach, issued Mar. 27, 1990; U.S. Pat. No. 4,855,090 to Wallach, issued Aug. 8, 1989; and U.S. Pat. No. 4,895,452 to Yioumas et al., issued Jan. 23, 1990.

[0057] For example, the term “nanocapsule” refers to a vesicular system or hollow particle with a shell surrounding a core-forming space, which, in certain instances, can be used for transporting a payload on a nanoscale level. A nanocapsule may also be a nano-sized version of a container. The payload of the nanocapsule can be, but is not limited to drugs, medicaments, pharmaceutical compositions, chemical compositions, therapeutic compositions, biological macromolecules, dyes, biological material, immunological compositions, nutritional compositions, vitamins, proteins, nucleic acids, antibodies, and vaccines. Various materials may be used for producing such nanocapsules. Nanocapsule refers to a particle having a hollow core that is surrounded by a shell, such that the particle has a size of less than about 1000 nanometers. When a nanocapsule includes a bioactive component, the bioactive component is located in the core that is surrounded by the shell of the nanocapsule.

[0058] As used herein, the term “nanocage” refers to a nanocapsule, whereby the shell is not solid, as described for the nanocapsule, but has multiple holes or pores in its shell, thereby making it possible for the payload within the core of the nanocage to come into contact with the surrounding environment. These holes or pores may be regular or irregular in shape and / or spacing on the surface of the particle.

[0059] The term “micelle,” a useful article in the employment of a general aspect of the present invention, can generally be thought of as a small - on the order of usually nanometers in diameter - aggregate of amphiphilic linear molecules having a polar, or hydrophilic end and an opposite non-polar, or hydrophobic end. These linear molecules can be comprised of simple molecules, or polymeric chains. A micelle can also be referred to as an aggregate of surfactant molecules dispersed in a liquid colloid. A typical micelle in aqueous solution can form an aggregate with the hydrophilic “head” regions in contact with surrounding solvent, and the sequestering of the hydrophobic tail regions in the micelle center. Other and similar definitions, descriptions and understandings of micelles are also known to those of skill in the art and are incorporated herein by reference.

[0060] The term “polymersome” as used herein refers to a vesicle-type which is typically composed of block copolymer amphiphiles, i.e., synthetic amphiphiles that have an amphiphilicity similar to that of lipids. By virtue of their amphiphilic nature (having a more hydrophilic block (head) and a more hydrophobic block (tail)), the block copolymers are capable of self-assembling into a head-to-tail and tail-to-head bilayer structure similar to liposomes. Compared to liposomes, polymersomes have much larger molecular weights, with number average molecular weights typically ranging from 1,000 to 100,000, from 2,500 to 50,000 or from 5,000 to 25,000, are typically chemically more stable, less leaky, less prone to interfere with biological membranes, and less dynamic due to a lower critical aggregation concentration. These properties result in less opsonisation and longer circulation times. The terms “more hydrophilic” and “more hydrophobic” as used in the context of the ampohiphilic nature of the block copolymers are used in a relative sense, i.e., both can be either hydrophilic or hydrophobic, as long as the difference in polarity between the blocks is sufficient for the formation of polymersomes according to the present invention. In view of the creation of a cavity in which water may be incorporated, in certain aspects the more hydrophilic end of the polymer is to be hydrophilic per se. Further, in view of the use as a therapeutic agent carrier, it is desired that also hydrophobic and / or hydrophilic therapeutic agents can be incorporated into the polymersomes. In one embodiment, the hydrophobic end of the polymer is hydrophobic per se. In one embodiment, the amphiphilic nature of the block copolymers is realized in the form of a block copolymer comprising a block made up of more hydrophilic monomeric units (A) and a block made up of more hydrophobic units (B), the block copolymer having the general structure AnBm, with n and m being integers of from 5 to 5,000, 10 to 1,000, or 10 to 500. It is also conceivable that at least one further units or blocks are built-in, e.g., a unit C with an intermediate hydrophilicity so as to yield a terpolymer having the general structure AnCpBm, with n and m being as defined above, and p being an integer of from 5 to 5,000, 10 to 1,000, or 10 to 500. Any of the blocks can itself be a copolymer, i.e., comprise different monomeric units of the required hydrophilic respectively hydrophobic nature. In one embodiment, the blocks themselves are homopolymeric. Any of the blocks, in particular the more hydrophilic block, may bear charges. The number and type of charges may depend on the pH of the environment. Any combination of positive and / or negative charges on any of the blocks is contemplated by the present invention.

[0061] “Dendrimers” have descriptions, definitions and understandings in the literature. For example, and without limitation and including other and similar definitions, descriptions and understandings in the art, the term dendrimer from the Greek word, “dendron,” for tree, can refer to a synthetic, three-dimensional molecule with branching parts. Descriptions and understandings of dendrimers can be gleaned from Holister et al., Dendrimers, Technology White Papers nr. 6, pub. October 2003 by cientifica, as well as the other literature published by those skilled in the art on dendrimers, all of which are incorporated herein by reference.

[0062] “Lamella” is a term whose definitions, descriptions and understandings are also known to those of skill in the art and which are incorporated herein by reference. In a very general sense, lamella or lamellae refers to plate-like, gill-shaped or other layered structures.

[0063] The definitions, descriptions and understandings of “nanovesicle” are well known to those of skill in the art, and are incorporated herein by reference. For example, “nanovesicle” can refer to a variety of small sac, sac-like or globular structures capable of containing fluid or other material therein.

[0064] As used herein, the term “exosome” refers to a subset of circulating microvesicles that are preformed microvesicles that are released from the cell following the exocytic fusion of intracellular multivesicular bodies with the plasma membrane, i.e., exosomes have an endocytic origin. As used herein, it is not intended that an exosome of the invention be limited by any particular size or size range. For example, exosomes include a variety of nanoparticles, including microvesicles, epididimosomes, argosomes, exosome-like vesicles, microparticles, promininosomes, prostasomes, dexosomes, texosomes, dex, tex, archeosomes and oncosomes. Exosomes are secreted by a wide range of cells, such as mammalian cells, and are secreted under both normal and pathological conditions. Exosomes, in some embodiments, function as intracellular messengers by virtue of carrying mRNA or other contents from a first cell to another cell (or plurality of cells). In some embodiments, exosomes are involved in blood coagulation, immune modulation, metabolic regulation, cell division, and other cellular processes.

[0065] The term “emulsion” as used herein refers to a mixture of two or more substances, such as liquids, which are normally immiscible, in which one substance forms droplets that are dispersed within another substance. One substance (the dispersed phase) is dispersed in the other (the continuous phase). Depending on the substances used, the droplets of an emulsion may be in the range of 1 nm to 100 pm, e.g., 1 nm to 100 nm, 1 pm to 50 pm, etc. For example, in one embodiment, the continuous phase is an aqueous phase, and the dispersed phase is an organic (oily or hydrophobic) phase; that is, the emulsion is an oil-in-water emulsion.

[0066] As used herein, the term “self-emulsifying drug delivery systems (SEDDS)” refers to isotropic mixtures, consisting of oils, surfactants, and / or cosolvents. In some embodiments, SEDDS increase solubility and bioavailability of poorly soluble drugs. For example, in some embodiments, designed SEDDS formulations are used to improve the oral absorption of highly lipophilic compounds. Examples of suitable SEDDS useful in the invention include, but are not limited to, those described in WO 2002 / 007712 A2 and Wang et al., 2014, J. Nutri. Biochem., 25:363-376, which are incorporated herein by reference in their entireties.

[0067] As used herein, the term “polymorph” refers to crystalline forms having the same chemical composition but different spatial arrangements of the molecules, atoms, and / or ions forming the crystal.

[0068] As used herein, the term “analog,” “analogue,” or “derivative” is meant to refer to a chemical compound or molecule made from a parent compound or molecule by at least one chemical reaction. As such, an analog can be a structure having a structure similar to that of the small molecule therapeutic agents described herein or can be based on a scaffold of a small molecule therapeutic agents described herein, but differing from it in respect to certain components or structural makeup, which may have a similar or opposite action metabolically. An analog or derivative can also be a small molecule that differs in structure from the reference molecule, but retains the essential properties of the reference molecule. An analog or derivative may change its interaction with certain other molecules relative to the reference molecule. An analog or derivative molecule may also include a salt, an adduct, tautomer, isomer, prodrug, or other variant of the reference molecule.

[0069] As used herein, the term “prodrug” refers to an agent that is converted into the parent drug in vivo. For example, the term “prodrug” refers to a derivative of a known direct acting drug, which derivative has enhanced delivery characteristics and therapeutic value as compared to the drug, and is transformed into the active drug by an enzymatic or chemical process. In some embodiments, “prodrug” refers to an inactive or relatively less active form of an active agent that becomes active by undergoing a chemical conversion through at least one metabolic process. In one embodiment, upon in vivo administration, a prodrug is chemically converted to the biologically, pharmaceutically, or therapeutically active form of the compound. In another embodiment, a prodrug is enzymatically metabolized by at least one step or processes to the biologically, pharmaceutically, or therapeutically active form of the compound. For example, the present compounds can be administered to a subject as a prodrug that includes an initiator bound to an active agent, and, by virtue of being degraded by a metabolic process, the active agent is released in its active form.

[0070] The term “tautomers” are constitutional isomers of organic compounds that readily interconvert by a chemical process (tautomerization).

[0071] The term “isomers” or “stereoisomers” refers to compounds, which have identical chemical constitution, but differ with regard to the arrangement of the atoms or groups in space.

[0072] As used herein, the term “stabilizers” refers to either, or both, primary particle and / or secondary stabilizers, which may be polymers or other small molecules. Non-limiting examples of primary particle and / or secondary stabilizers for use with the present invention include, e.g., starch, modified starch, and starch derivatives, gums, including but not limited to polymers, polypeptides, albumin, amino acids, thiols, amines, carboxylic acid, and any combinations or derivatives thereof. Other examples include, but are not limited to, xanthan gum, alginic acid, other alginates, benitoniite, veegum, agar, guar, locust bean gum, gum arabic, quince psyllium, flax seed, okra gum, arabinoglactin, pectin, tragacanth, scleroglucan, dextran, amylose, amylopectin, dextrin, etc., cross-linked polyvinylpyrrolidone, ion-exchange resins, potassium polymethacrylate, carrageenan (and derivatives), gum karaya, and biosynthetic gum. Other examples of useful primary particle and / or secondary stabilizers include polymers, such as polycarbonates (linear polyesters of carbonic acid); microporous materials (bisphenol, a microporous poly(vinylchloride), micro-porous polyamides, microporous modacrylic copolymers, microporous styrene-acrylic and its copolymers); porous polysulfones, halogenated poly(vinylidene), polychloroethers, acetal polymers, polyesters prepared by esterification of a dicarboxylic acid or anhydride with an alkylene polyol, poly(alkylenesulfides), phenolics, polyesters, asymmetric porous polymers, cross-linked olefin polymers, hydrophilic microporous homopolymers, copolymers or interpolymers having a reduced bulk density, and other similar materials, poly(urethane), cross-linked chain-extended poly(urethane), poly(amides), poly(imides), poly(benzimidazoles), collodion, regenerated proteins, semi-solid cross-linked poly(vinylpyrrolidone).

[0073] The terms “coat,” “coated,” or “coating,” as used herein, refer to at least a partial coating of the nanoparticle. One hundred percent coverage is not necessarily implied by these terms.

[0074] An “effective amount” or “therapeutically effective amount,” as used herein, means an amount, which provides a therapeutic or prophylactic benefit. For example, an “effective amount” or “therapeutically effective amount” of a nanoparticle is that amount of compound, which is sufficient to provide a beneficial effect (e.g., deliver a nutritional or bioactive agent) to the subject to which the nanoparticle is administered.

[0075] “Pharmaceutically acceptable” refers to those properties and / or substances, which are acceptable to the subject from a pharmacological / toxicological point of view and to the manufacturing pharmaceutical chemist from a physical / chemical point of view regarding composition, formulation, stability, subject acceptance and bioavailability. “Pharmaceutically acceptable carrier” refers to a medium that does not interfere with the effectiveness of the biological activity of the active ingredient(s) and is not toxic to the host to which it is administered.

[0076] As used herein, the term “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the invention within or to the subject such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound useful within the invention, and not injurious to the subject. Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as com starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations. As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, anti-bacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound useful within the invention, and are physiologically acceptable to the subject. Supplementary active compounds may also be incorporated into the compositions. The “pharmaceutically acceptable carrier” may further include a pharmaceutically acceptable salt of the compound useful within the invention. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the invention are known in the art.

[0077] The term “pharmaceutically acceptable salt” refers to any pharmaceutically acceptable salt, which upon administration to the subject is capable of providing (directly or indirectly) a compound as described herein. In some embodiments, such salts are acid addition salts with physiologically acceptable organic or inorganic acids. Examples of the acid addition salts include mineral acid addition salts such as, for example, hydrochloride, hydrobromide, hydroiodide, sulphate, nitrate, phosphate, and organic acid addition salts, such as, for example, acetate, trifluoroacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, malate, mandelate, methane sulphonate, and p-toluenesulphonate. Examples of the alkali addition salts include inorganic salts, such as, for example, sodium, potassium, calcium and ammonium salts, and organic alkali salts, such as, for example, ethylenediamine, ethanolamine, N,N- dialkylenethanolamine, triethanolamine, and basic amino acids salts. However, it will be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the invention since those may be useful in the preparation of pharmaceutically acceptable salts. Procedures for salt formation are conventional in the art.

[0078] The term “solvate” in accordance with this invention should be understood as meaning any form of the active compound in accordance with the invention in which the said compound is bonded by a non-covalent bond to another molecule (normally a polar solvent), including especially hydrates and alcoholates.

[0079] The term “pharmacological composition,” “therapeutic composition,” “therapeutic formulation” or “pharmaceutically acceptable formulation” can mean, but is in no way limited to, a composition or formulation that allows for the effective distribution of an agent provided by the invention, which is in a form suitable for administration to the physical location most suitable for their desired activity, e.g., systemic administration. The pharmaceutical composition facilitates administration of the compound to an organism. Multiple techniques of administering a compound exist in the art including, but not limited to, intravenous, oral, aerosol, parenteral, ophthalmic, pulmonary and topical administration.

[0080] The term “therapeutic” as used herein means a treatment and / or prophylaxis. A therapeutic effect is obtained by suppression, diminution, remission, or eradication of at least one sign or symptom of a disease or disorder state.

[0081] As used herein, the terms “therapeutic compound,” “therapeutic agent”, “drug”, “active pharmaceutical”, and “active pharmaceutical ingredient” are used interchangeably to refer to chemical entities that display certain pharmacological effects in a body and are administered for such purpose. Non-limiting examples of therapeutic agents include, but are not limited to, hydrophilic therapeutic agents, hydrophobic therapeutic agents, antibiotics, antibodies, small molecules, anti-cancer agents, chemotherapeutic agents, immunomodulatory agents, RNA molecules, siRNA molecules, DNA molecules, gene editing agents, gene-silencing agents, CRISPR-associated agents (e.g., guide RNA molecules, endonucleases, and variants thereof), analgesics, vaccines, anticonvulsants; anti-diabetic agents, antifungal agents, antineoplastic agents, anti-parkinsonian agents, anti-rheumatic agents, appetite suppressants, biological response modifiers, cardiovascular agents, central nervous system stimulants, contraceptive agents, dietary supplements, vitamins, minerals, lipids, saccharides, metals, amino acids (and precursors), nucleic acids and precursors, contrast agents, diagnostic agents, dopamine receptor agonists, erectile dysfunction agents, fertility agents, gastrointestinal agents, hormones, immunomodulators, anti-hypercalcemia agents, mast cell stabilizers, muscle relaxants, nutritional or bioactive agents, ophthalmic agents, osteoporosis agents, psychotherapeutic agents, parasympathomimetic agents, parasympatholytic agents, respiratory agents, sedative hypnotic agents, skin and mucous membrane agents, smoking cessation agents, steroids, sympatholytic agents, urinary tract agents, uterine relaxants, vaginal agents, vasodilator, anti-hypertensive, hyperthyroids, anti-hyperthyroids, anti-asthmatics and vertigo agents. In certain embodiments, the at least one therapeutic agents is water-soluble, poorly water-soluble drug or a drug with a low, medium, or high melting point. The therapeutic agents may be provided with or without a stabilizing salt or salts.

[0082] Some examples of active ingredients suitable for use in the pharmaceutical formulations and methods of the present invention include: hydrophilic, lipophilic, amphiphilic or hydrophobic, and that can be solubilized, dispersed, or partially solubilized and dispersed, on or about the nanocluster. The active agent-nanocluster combination may be coated further to encapsulate the agent-nanocluster combination and may be directed to a target by functionalizing the nanocluster with, e.g., aptamers and / or antibodies. Alternatively, an active ingredient may also be provided separately from the solid pharmaceutical composition, such as for coadministration. Such active ingredients can be any compound or mixture of compounds having therapeutic or other value when administered to an animal, particularly to a mammal, such as drugs, nutrients, cosmeceuticals, nutraceuticals, diagnostic agents, nutritional or bioactive agents, and the like. The active agents described herein may be found in their native state, however, they will generally be provided in the form of a salt. The active agents described herein include their isomers, analogs, and derivatives.

[0083] A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate.

[0084] In contrast, a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal’s state of health.

[0085] The terms “patient,” “subject,” “individual,” and the like are used interchangeably herein, and refer to any animal, or cells thereof whether in vitro or in situ, amenable to the methods described herein. In certain non-limiting embodiments, the patient, subject or individual is a human.

[0086] To “treat” a disease as the term is used herein, means to reduce the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject.

[0087] As used herein, “treating a disease or disorder” means reducing the frequency with which a symptom of the disease or disorder is experienced by a patient. Disease and disorder are used interchangeably herein.

[0088] A disease or disorder is “alleviated” if the severity of a sign or symptom of the disease or disorder, the frequency with which such a sign or symptom is experienced by a patient, or both, is reduced.

[0089] By the term “modulating,” as used herein, is meant mediating a detectable increase or decrease in the level of a response in a subject compared with the level of a response in the subject in the absence of a treatment or compound, and / or compared with the level of a response in an otherwise identical but untreated subject. The term encompasses perturbing and / or affecting a native signal or response thereby mediating a beneficial therapeutic response in a subject.

[0090] Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0091] Description

[0092] The present invention is based, in part, on the unexpected discovery that nanoparticles comprising resveratrol and quercetin had synergistically increased anti-obesity activities. Thus, in one aspect, the present invention provides nanoparticles or compositions thereof comprising resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof, at least one lipid, at least one surfactant, and at least one vitamin E. In some embodiments, the nanoparticles or compositions thereof further comprise at least one targeting agent. In some embodiments, the nanoparticles or compositions thereof are targeted for an adipose tissue of interest. In some embodiments, the nanoparticles or compositions thereof further comprise at least one coating agent.

[0093] In some aspects, the present invention provides methods of activating brown adipose tissue of interest. In other aspects, the present invention provides methods of browning white adipose tissue and / or beige adipose tissue of interest. In various aspects, the present invention also provides methods of increasing the concentration of a cellular cAMP.

[0094] In some aspects, the present invention provides methods of reducing body fat, inducing weight loss, and / or improving glucose homeostasis and insulin sensitivity. In some aspects, the present invention provides methods of treating or preventing a metabolic disease or disorder, such as obesity, obesity-related disease or disorder, diabetes, fatty liver disease, nonalcoholic fatty liver disease, cardiovascular disease, etc.

[0095] Composition of Nanoparticles

[0096] The present invention relates, in part, to a nanoparticle comprising resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof. In some embodiments, the nanoparticle further comprises at least one lipid, at least one surfactant, and at least one vitamin E. In various embodiments, the nanoparticle selectively targets at least one adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.). In various embodiments, the nanoparticle selectively binds at least one adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.).

[0097] In various embodiments, the nanoparticle increases the concentration of a cellular cAMP in an adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.), induces browning of an adipose tissue of interest (e.g., WAT, beige adipose tissue, etc.), activates an adipose tissue of interest (e.g., BAT, etc.), increases the level or activity of a 5’ adenosine monophosphate-activated protein kinase (AMPK), increases the level or activity sirtuin 1 (SIRT1), increases the level or activity peroxisome proliferator-activated receptor gamma coacvtivator 1-alpha (PGC-la), decreases the level or activity of a fatty acid synthase (FAS), decreases the level or activity of acetyl-CoA carboxylase (ACC), decreases the level or activity of NF-KB, decreases the level or activity of interferon-y, decreases the level or activity of IL-4, decreases the level or activity of IL- la, decreases the level or activity of a phosphodiesterase (PDE), or a combination thereof. In some embodiments, the nanoparticle increases mitochondrial biogenesis, increases mitochondrial function and thermogenesis, increases gut integrity, increases gut health, increases insulin sensitivity, increases energy expenditure of at least one cell of interest, improves metabolic health, improves gut microbiota profde, improves glucose homeostasis, improves lipid homeostasis, decreases fat mass and body weight, decreases inflammatory response, decreases liver damage, or a combination thereof. In some embodiments, the nanoparticle reduces body fat in a subject. In some embodiments, the nanoparticle induces weight loss in a subject. In some embodiments, the nanoparticle does not change the level or concentration of lean mass.

[0098] In another aspect, the present invention provides a composition comprising resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, at least one lipid, at least one surfactant, and at least one vitamin E.

[0099] In various embodiments, the composition increases the concentration of a cellular cAMP in an adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.), induces browning of an adipose tissue of interest (e.g., WAT, beige adipose tissue, etc.), activates an adipose tissue of interest (e.g., BAT, etc.), increases the level or activity of a 5’ adenosine monophosphate-activated protein kinase (AMPK), increases the level or activity of sirtuin 1 (SIRT1), increases the level or activity of peroxisome proliferator-activated receptor gamma coacvtivator 1-alpha (PGC-la), decreases the level or activity of a fatty acid synthase (FAS), decreases the level or activity of acetyl-CoA carboxylase (ACC), decreases the level or activity of NF-KB, decreases the level or activity of interferon-^, decreases the level or activity of IL-4, decreases the level or activity of IL- la, decreases the level or activity of a phosphodiesterase (PDE), or a combination thereof. In some embodiments, the composition increases mitochondrial biogenesis, increases mitochondrial function and thermogenesis, increases gut integrity, increases gut health, increases insulin sensitivity, increases energy expenditure of at least one cell of interest, improves metabolic health, improves gut microbiota profile, improves glucose homeostasis, improves lipid homeostasis, decreases fat mass and body weight, decreases inflammatory response, decreases liver damage, or a combination thereof. In some embodiments, the composition reduces body fat in a subject. In some embodiments, the composition induces weight loss in a subject. In some embodiments, the composition does not change the level or concentration of lean mass.

[0100] In various embodiments, resveratrol, or the derivative, prodrug, or pharmaceutically acceptable salt thereof, is encapsulated within the nanoparticle, adhered to the surface of the nanoparticle, integrated into the structure of the nanoparticle, bound to the nanoparticle, or any combination thereof. In one embodiment, resveratrol, or the derivative, prodrug, or pharmaceutically acceptable salt thereof, coats the outside surface of the nanoparticle. In one embodiment, resveratrol, or the derivative, prodrug, or pharmaceutically acceptable salt thereof, coats a portion of the outside surface of the nanoparticle.

[0101] In various embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 0.01 wt% to about 99.99 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 0.1 wt% to about 99.9 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 60 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 55 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 0.1 wt% to about 50 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 50 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 10 wt%.

[0102] For example, in some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 0.01 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 0.02 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 0.05 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 0.15 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 1.0 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 7 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 10 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 20 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 30 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 40 wt%. In some embodiments, the nanoparticle or composition comprises resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 50 wt%.

[0103] In various embodiments, quercetin, or the derivative, prodrug, or pharmaceutically acceptable salt thereof, is encapsulated within the nanoparticle, adhered to the surface of the nanoparticle, integrated into the structure of the nanoparticle, bound to the nanoparticle, or any combination thereof. In one embodiment, quercetin, or the derivative, prodrug, or pharmaceutically acceptable salt thereof, coats the outside surface of the nanoparticle. In one embodiment, quercetin, or the derivative, prodrug, or pharmaceutically acceptable salt thereof, coats a portion of the outside surface of the nanoparticle.

[0104] In various embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 0.01 wt% to about 99.99 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 0.1 wt% to about 99.9 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 60 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 55 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 0.1 wt% to about 50 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 50 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration range of about 1 wt% to about 10 wt%.

[0105] For example, in some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 0.01 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 0.02 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 0.05 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 0.15 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 1.0 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 7 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 10 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 20 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 30 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 40 wt%. In some embodiments, the nanoparticle or composition comprises quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, in a concentration of about 50 wt%.

[0106] In some embodiments, the lipid is selected from a fatty acid, wax, sterol, lipid- soluble vitamin, such as vitamins A, D, E, and K, monoglyceride, diglyceride, triglyceride, phospholipids, cholesterol, PEG-phospholipids, lipid conjugates, or any combination thereof. In some embodiments, the phospholipid is selected from phosphatidylcholine (PC), such as soy or egg L-a-phosphatidylcholine (PC), phosphatidic acid (PA), phosphatidylethanolamine (cephalin) (PE), lecithin, phosphatidyl serine (PS), phosphoinositide, phosphatidylinositol (PI), phosphatidylinositol phosphate (PIP), phosphatidylinositol bisphosphate (PIP2), phosphatidylinositol trisphosphate (PIP3), phosphosphingolipid, phospholipid-phosphatic acid, sphingolipid, ceramide phosphorylcholine (sphingomyelin) (SPH), ceramide phosphorylethanolamine (sphingomyelin) (Cer-PE), ceramide phosphoryl lipid, or any combination thereof.

[0107] In various embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 0.1 wt% to about 100 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 1 wt% to about 100 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 5 wt% to about 90 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 1 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 10 wt% to about 80 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 10 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 10 wt% to about 50 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 15 wt% to about 45 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration range of about 35 wt% to about 40 wt%.

[0108] For example, in some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 1 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 2 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 5 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 5.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 10 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 13 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 15 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 20 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 25 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 30 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 35 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 37 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 40 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 45 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 50 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 60 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 80 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 90 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 95 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 95.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 99 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 99.9 wt%. In some embodiments, the nanoparticle or composition comprises at least one lipid in a concentration of about 100 wt%.

[0109] In some embodiments, the surfactant is any surfactant that is known to those of skill in the art, any emulsifier that is known to those of skill in the art, or any combination thereof. Examples of such surfactants include, but are not limited to, polyethylene glycol (PEG), functionalized PEG, such as polyethylene glycol 15 -hydroxy stearate (Kolliphor® HS15), POLYOXYL (40) STEARATE, poly(ethylene glycol)-100-stearate (Myrj59), poly(ethylene glycol)-40-stearate (Myrj 52), Tween® 80 (polysorbate 80), mustard, lecithin, such as soy lecithin and egg lecithin, monoglycerides, diglycerides, polysorbates, carrageenan, guar gum, canola oil, polysorbates (Tween™), sodium dodecyl sulfate (sodium lauryl sulfate), lauryl dimethyl amine oxide, cetyltrimethylammonium bromide (CTAB), polyethoxylated alcohols, polyoxyethylene sorbitan, octoxynol (Triton XI 00™), N, N-dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide (HTAB), polyoxyl 10 lauryl ether, Brij 721™ (poly(ethylene glycol) 21 -stearate), bile salts, such as sodium deoxycholate and sodium cholate, polyoxyl castor oil (Cremophor™), nonylphenol ethoxylate (Tergitol™), cyclodextrins, methylbenzethonium chloride (Hyamine™), or a combination thereof. Examples of such emulsifier include, but are not limited to emulsifiers listed in “Emulsifiers: Types and Uses, R Miller, Kansas State University, Manhattan, KS, USA, 2016 Elsevier Ltd. All rights reserved,” incorporated herein by reference in its entirety.

[0110] In various embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 0.1 wt% to about 100 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 1 wt% to about 100 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 1 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 10 wt% to about 80 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 10 wt% to about 75 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 10 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 10 wt% to about 50 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 15 wt% to about 45 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration range of about 35 wt% to about 40 wt%.

[0111] For example, in some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 1 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 2 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 5 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 5.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 10 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 13 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 15 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 20 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 25 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 30 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 35 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 37 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 40 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 45 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 50 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 60 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 80 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 90 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 95 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 95.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 99 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 99.9 wt%. In some embodiments, the nanoparticle or composition comprises at least one surfactant in a concentration of about 100 wt%.

[0112] In some embodiments, the vitamin E is selected from a-tocopherol, a-tocopherol acetate (aTA), a-tocopherol nicotinate, D-a-tocopheryl polyethylene glycol 1000 succinate, - tocopherol, P-tocopherol acetate, P-tocopherol nicotinate, y-tocopherol, y-tocopherol acetate, y- tocopherol nicotinate, 5-tocopherol, 6-tocopherol acetate, 6-tocopherol nicotinate, a-tocotrienol, a-tocotrienol acetate, a-tocotrienol nicotinate, P-tocotrienol, P-tocotrienol acetate, P-tocotrienol nicotinate, y-tocotrienol, y-tocotrienol acetate, y-tocotrienol nicotinate, 6-tocotrienol, 8- tocotrienol acetate, 6-tocotrienol nicotinate, and any combination thereof.

[0113] In various embodiments, the nanoparticle or composition comprises at least one vitamin E in a concentration range of about 0.1 wt% to about 100 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration range of about 1 wt% to about 100 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration range of about 1 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration range of about 5 wt% to about 75 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration range of about 10 wt% to about 80 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration range of about 10 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration range of about 10 wt% to about 50 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration range of about 15 wt% to about 45 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration range of about 35 wt% to about 40 wt%.

[0114] For example, in some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 1 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 2 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 5 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 5.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 10 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 13 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 15 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 20 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 25 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 30 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 35 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 37 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 40 wt%. Tn some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 45 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 50 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 60 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 80 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 90 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 95 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 95.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 99 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 99.9 wt%. In some embodiments, the nanoparticle or composition comprises at least one vitamins E in a concentration of about 100 wt%.

[0115] In various embodiments, the nanoparticle or composition further comprises at least one targeting agent that selectively targets an adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.). In various embodiments, the targeting agent is encapsulated within the nanoparticle, adhered to the surface of the nanoparticle, integrated into the structure of the nanoparticle, bound to the nanoparticle, or any combination thereof. In one embodiment, the targeting agent coats the outside surface of the nanoparticle. In one embodiment, the targeting agent coats a portion of the outside surface of the nanoparticle. Examples of such targeting agents include, but are not limited to, proteins, peptides, antibodies, lipids, polysaccharides, nucleic acids, aptamers, and small molecules. In some embodiments, the targeting agent is a peptide of SEQ ID NO: 1 (CSWKYWFGEC) or SEQ ID NO:2 (GSWKYWFGEGGC).

[0116] In various embodiments, the nanoparticle comprises the targeting agent in a concentration range of about 0.01 mol% to about 99.99 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration range of about 0.1 mol% to about 99.9 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration range of about 1 mol% to about 70 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration range of about 1 mol% to about 60 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration range of about 1 mol% to about 55 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration range of about 1 mol% to about 50 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration range of about 10 mol% to about 50 mol%.

[0117] For example, in some embodiments, the nanoparticle comprises the targeting agent in a concentration of about 0.01 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration of about 0.02 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration of about 0.05 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration of about 0.15 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration of about 1.0 mol%. In some embodiments, the nanoparticle comprises the targeting agent in a concentration of about 10 mol%.

[0118] In various embodiments, the nanoparticle or composition further comprises at least one helper compound. In some embodiments, the helper compound is a helper lipid, helper polymer, or any combination thereof. In some embodiments, the helper lipid is phospholipid, cholesterol lipid, polymer, cationic lipid, neutral lipid, charged lipid, steroid, steroid analogue, polymer conjugated lipid, stabilizing lipid, or any combination thereof.

[0119] In some embodiments, the phospholipid is dioleoyl-phosphatidylethanolamine (DOPE) or a derivative thereof, distearoylphosphatidylcholine (DSPC) or a derivative thereof, distearoyl-phosphatidylethanolamine (DSPE) or a derivative thereof, stearoyloleoylphosphatidylcholine (SOPC) or a derivative thereof, l-stearioyl-2-oleoyl- phosphatidyethanol amine (SOPE) or a derivative thereof, N-(2,3-dioleoyloxy)propyl)-N,N,N- trimethylammonium chloride (DOTAP) or a derivative thereof, or any combination thereof.

[0120] In some embodiments, the cholesterol lipid is cholesterol or a derivative thereof, such as a substituted cholesterol molecule. In some embodiments, the nanoparticle or composition comprises a mixture of cholesterol and a substituted cholesterol molecule.

[0121] In some embodiments, the polymer is polyethylene glycol (PEG) or a derivative thereof.

[0122] As used herein, the term “cationic lipid” refers to a lipid that is cationic or becomes cationic (protonated) as the pH is lowered below the pK of the ionizable group of the lipid, but is progressively more neutral at higher pH values. At pH values below the pK, the lipid is then able to associate with negatively charged nucleic acids. In certain embodiments, the cationic lipid comprises a zwitterionic lipid that assumes a positive charge on pH decrease.

[0123] In some embodiments, the cationic lipid comprises any of a number of lipid species which carry a net positive charge at a selective pH, such as physiological pH. Such lipids include, but are not limited to, N,N-dioleyl-N,N-dimethylammonium chloride (DODAC); N- (2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA); N,N-distearyl-N,N- dimethylammonium bromide (DDAB); N-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP); 3-(N-(N',N'-dimethylaminoethane)-carbamoyl)cholesterol (DC-Chol), N-(l- (2,3-dioleoyloxy)propyl)-N-2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoracetate (DOSPA), dioctadecylamidoglycyl carb oxy spermine (DOGS), l,2-dioleoyl-3- dimethylammonium propane (DODAP), N,N-dimethyl-2,3-dioleoyloxy)propylamine (DODMA), and N-(l,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide (DMRIE). Additionally, a number of commercial preparations of cationic lipids are available which can be used in the present invention. These include, for example, LIPOFECTIN® (commercially available cationic liposomes comprising DOTMA and 1,2- dioleoyl-sn-3-phosphoethanolamine (DOPE), from GIBCO / BRL, Grand Island, N.Y.); LIPOFECTAMINE® (commercially available cationic liposomes comprising N-(l-(2,3- dioleyloxy)propyl)-N-(2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoroacetate (DOSPA) and (DOPE), from GIBCO / BRL); and TRANSFECTAM® (commercially available cationic lipids comprising dioctadecylamidoglycyl carboxyspermine (DOGS) in ethanol from Promega Corp., Madison, Wis.). The following lipids are cationic and have a positive charge at below physiological pH: DODAP, DODMA, DMDMA, l,2-dilinoleyloxy-N,N- dimethylaminopropane (DLinDMA), 1 ,2-dilinolenyloxy-N,N-dimethylaminopropane (DLenDMA).

[0124] In one embodiment, the cationic lipid is an amino lipid. Suitable amino lipids useful in the invention include those described in WO 2012 / 016184, incorporated herein by reference in its entirety. Representative amino lipids include, but are not limited to, 1,2- dilinol ey oxy-3 -(dimethylamino)acetoxypropane (DLin-DAC), 1,2-dilinoley oxy-3 - morpholinopropane (DLin-MA), l,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2- dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1 -linoleoyl-2-linoleyloxy-3- dimethylaminopropane (DLin-2-DMAP), l,2-dilinoleyloxy-3 -trimethylaminopropane chloride salt (DLin-TMA.Cl), l,2-dilinoleoyl-3 -trimethylaminopropane chloride salt (DLin-TAP.Cl), 1,2- dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), 3-(N,N-dilinoleylamino)-l,2- propanediol (DLinAP), 3-(N,N-dioleylamino)-l,2-propanediol (DOAP), l,2-dilinoleyloxo-3-(2- N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), and 2,2-dilinoleyl-4- dimethylaminomethyl-[l,3]-di oxolane (DLin-K-DMA).

[0125] In one embodiment, the lipid is a PEGylated lipid, including, but not limited to, DSPE-PEG-DBCO, DOPE-PEG-Azide, DSPE-PEG- Azide, DPPE-PEG- Azide, DSPE-PEG- Carboxy-NHS, DOPE-PEG-Carboxylic Acid, DSPE-PEG-Carboxylic acid.

[0126] The term “neutral lipid” refers to any one of a number of lipid species that exist in either an uncharged or neutral zwitterionic form at physiological pH. Representative neutral lipids include diacylphosphatidylcholines, diacylphosphatidylethanolamines, ceramides, sphingomyelins, dihydro sphingomyelins, cephalins, and cerebrosides.

[0127] Exemplary neutral lipids include, for example, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE) and dioleoyl-phosphatidylethanolamine 4- (N-maleimidomethyl)-cyclohexane-l -carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoylphosphatidylethanolamine (DSPE), distearoyl-phosphatidylethanolamine (DSPE)-maleimide- PEG, distearoyl-phosphatidylethanolamine (DSPE)-maleimide-PEG2000, 16-0-monom ethyl PE, 16-O-dimethyl PE, 18-1 -trans PE, l-stearioyl-2-oleoyl-phosphatidy ethanol amine (SOPE), stearoyloleoylphosphatidylcholine (SOPC), and l,2-dielaidoyl-sn-glycero-3- phophoethanolamine (transDOPE). In one embodiment, the neutral lipid is 1,2-distearoyl-sn- glycero-3 -phosphocholine (DSPC).

[0128] In some embodiments, the composition comprises a neutral lipid selected from DSPC, DPPC, DMPC, DOPC, POPC, DOPE, or SM.

[0129] A “steroid” is a compound comprising the following carbon skeleton:

[0130] In certain embodiments, the steroid or steroid analogue is cholesterol. In some of these embodiments, the molar ratio of the cationic lipid.

[0131] The term “anionic lipid” refers to any lipid that is negatively charged at physiological pH. These lipids include phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoylphosphatidylethanolamines, N- succinylphosphatidylethanolamines, N-glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids.

[0132] The term “polymer conjugated lipid” refers to a molecule comprising both a lipid portion and a polymer portion. An example of a polymer conjugated lipid is a pegylated lipid. The term “pegylated lipid” refers to a molecule comprising both a lipid portion and a polyethylene glycol portion. Pegylated lipids are known in the art and include polyethylene glycol (PEG), maleimide PEG (mPEG), DSPE-PEG-DBCO, l-(m on om ethoxy-poly ethyl eneglycol)-2, 3 -dimyristoylglycerol (PEG-s- DMG), DOPE-PEG- Azide, DSPE-PEG- Azide, DPPE-PEG- Azide, DSPE-PEG-Carboxy-NHS, DOPE-PEG- Carboxylic Acid, DSPE-PEG-Carboxylic acid and the like.

[0133] In certain embodiments, the stabilizing lipid is a polyethylene glycol-lipid (pegylated lipid). Suitable polyethylene glycol-lipids include PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramides (e.g., PEG-CerC14 or PEG-CerC20), PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols. Representative polyethylene glycol-lipids include PEG-c- DOMG, PEG-c-DMA, and PEG-s-DMG. In one embodiment, the polyethylene glycol-lipid is N- [(methoxy poly(ethylene glycol)2ooo)carbamyl]-l,2-dimyristyloxlpropyl-3-amine (PEG-c-DMA). In one embodiment, the polyethylene glycol-lipid is PEG-c-DOMG). In other embodiments, the nanoparticles comprise a pegylated diacylglycerol (PEG-DAG) such as l-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG-DMG), a pegylated phosphatidylethanoloamine (PEG-PE), a PEG succinate diacylglycerol (PEG-S-DAG) such as 4- O-(2’,3’-di(tetradecanoyloxy)propyl-l-O-(<n-methoxy(polyethoxy)ethyl)butanedioate (PEG-S- DMG), a pegylated ceramide (PEG-cer), or a PEG dialkoxypropylcarbamate such as co- methoxy(polyethoxy)ethyl-N-(2,3-di(tetradecanoxy)propyl)carbamate or 2,3- di(tetradecanoxy)propyl-N-(w-methoxy(polyethoxy)ethyl)carbamate.

[0134] In various embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 0 wt% to about 100 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 0.01 wt% to about 99.99 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 0.1 wt% to about 99.9 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 0.1 wt% to about 90 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 0.1 wt% to about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 5 wt% to about 95 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 0.5 wt% to about 50 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 0.5 wt% to about 47 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration range of about 2.5 wt% to about 47 wt%.

[0135] For example, in some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 0.01 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 0.1 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 0.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 1 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 1.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 2 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 2.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 5 wt%. Tn some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 10 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 12 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 15 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 16 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 20 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 25 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 30 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 35 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 37 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 40 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 45 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 46.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 47 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 50 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 60 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 63 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 70 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 80 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 90 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 95 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 95.5 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 99 wt%. In some embodiments, the nanoparticle or composition comprises at least one helper compound in a concentration of about 100 wt%.

[0136] In various aspects, the nanoparticle comprises at least one stabilizer.

[0137] In one embodiment, the nanoparticle or composition is a polymersome. Any polymersome known in the art may be utilized. Thus, in one embodiment, the nanoparticle or composition comprises a homopolymer. In some embodiments, the nanoparticle or composition comprises a block copolymer that is a triblock, tetrablock, pentablock, or at least six block copolymer. In some embodiments, the nanoparticle or composition comprises polyethylene oxide) (PEO) block copolymer, poly(ethylethylene) (PEE), poly (butadiene) (PB or PBD), poly(styrene) (PS), poly (isoprene) (PI), PEI, poly(lactide-co-glycolic acid) (PLGA), biodegradable PLGA, polyethylene glycol (PEG), poly(lactide-co-glycolic acid)-polyethylene glycol (PLGA-PEG), poly(lactide-co-glycolic acid)-block-polyethylene glycol (PLGA-b-PEG), biodegradable PLGA-PEG, biodegradable PLGA-b-PEG, polyanhydride, polyanhydride-block- PEG copolymers, zwitterionic poly(carbobetaine), zwitterionic poly(sulfobetaine)-containing, zwitterionic poly(carbobetaine) and zwitterionic poly(sulfobetaine)-containing copolymers, poly(acrylic acid-co-distearin acrylate), poly(trimethylene carbonate)-block-poly(L-gluatamic acid), poly(ethylene glycol-block-L-aspartic acid), poly(2-hydroxyethyl-co-octadecyl aspartamide), poly(ethylene glycol-co-trimethylene carbonate-co-caprolactone, polypropylene oxide block copolymers, polyethylene oxide-block-polypropylene oxide copolymers, or any combination thereof.

[0138] In some embodiments, the nanoparticle or composition comprises poly(s- caprolactone) (PCL) diblock co-polymer. In some embodiments, the nanoparticle or composition comprises poly(ethylene oxide)-block-poly(s-caprolactone) (PEO-b-PCL) based diblock copolymers. In some embodiments, the nanoparticle or composition is derived from the coupling of poly(lactic acid), poly(glycolide), poly(lactic-coglycolic acid) and / or or poly(3- hydroxybutyrate) with PEO. In some embodiments, the nanoparticle or composition comprises PLGA. In some embodiments, the nanoparticle or composition comprises PEG. In one embodiment, the nanoparticle or composition comprises poly(lactide-co-glycolic acid)- polyethylene glycol (PLGA-PEG). For example, in some embodiments, a PLGA-PEG polymersome encapsulates the ICGJ and optionally PEI.

[0139] In one embodiment, the nanoparticle further comprises a cationic polymer. The cationic polymer may be a straight chain polymer (i.e., linear polymer) or a branched chain polymer (i.e., branched polymer), including hyperbranched polymers. In one embodiment the cationic polymer is a branched cationic polymer. In one embodiment, the cationic polymer is cross-linked. In one embodiment, the cationic polymer is a polyamine. In one embodiment, the cationic polymer has a molecular weight of 5 kDa-3000 kDa. For example, in one embodiment, the cationic polymer has a molecular weight of 5 kDa-2000 kDa, 5 kDa-1500 kDa, 5 kDa-1000 kDa, 5 kDa-800 kDa, 5 kDa-500 kDa, 5 kDa-300 kDa or 5 kDa-200 kDa or 800 kDa-3000 kDa.

[0140] In one embodiment, the cationic polymer is a polyalkyleneimine (e g., polyethyleneimine), polyallylamine, polyamidoamine, or poly(amino-co-ester). In one embodiment, the cationic polymer is polyethyleneimine (PEI), chitosan, poly(2-N,N- dimethylaminoethylmethacrylate), or poly-L-lysine. In one embodiment, the cationic polymer stabilizes the nanocapsule.

[0141] In various embodiments, the nanoparticle or composition further comprises at least one phytochemical or bioactive compound selected from the group consisting of curcumin, theaflavins, thearubigins, epigallocatechin gallate (EGCG), (+)-catechin, (-)-epicatechin, (-)- epicatechin gallate, (-)-epigallocatechin, (+)-gallocatechin, isorhamnetin, kaempferol, myricetin, apigenin, luteolin, baicalein, chrysin, forskolin, chlorophyll a, chlorophyll b, eriodictyol, hesperetin, naringenin, taxifolin, catechins, luteolin, cyanidin, genistein, daidzein, genistein, glycitein, biochanin A, formononetin, cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, proanthocyanidins, a-carotene, P-carotene, P-cryptoxanthin, lutein, zeaxanthin, lycopene, and any combination thereof.

[0142] In various embodiments, the nanoparticle or composition further comprises at least one coating agent. In one embodiment, the coating agent coats the outside surface of the nanoparticle. In one embodiment, the coating agent coats a portion of the outside surface of the nanoparticle. In some embodiments, the coating agent comprises a starch, stabilizer, plasticizer, lipid, polymer, polysaccharide, protein, zein, soy protein, whey, casein, fatty acid, wax, neutral lipid, resin, cellulose, chitosan, alginate, starch, or any combination thereof.

[0143] In one embodiment, the coating agent comprises a biocompatible polymer. Examples of coating agents include, but are not limited to, biocompatible polymer, a biodegradable polymer, a multifunctional linker, starch, modified starch, and starch derivatives, gums, including but not limited to polymers, polypeptides, albumin, amino acids, alcohols (e.g., PVA, ethyl alcohol, etc.), thiols, amines, carboxylic acid and combinations or derivatives thereof, citric acid, xanthan gum, alginic acid, other alginates, benitoniite, veegum, agar, guar, locust bean gum, gum arabic, quince psyllium, flax seed, okra gum, arabinoglactin, pectin, tragacanth, scleroglucan, dextran, amylose, amylopectin, dextrin, etc., cross-linked polyvinylpyrrolidone, ion-exchange resins, potassium polymethacrylate, carrageenan (and derivatives), gum karaya and biosynthetic gum, polycarbonates (linear polyesters of carbonic acid); microporous materials (bisphenol, a microporous poly(vinylchloride), micro-porous polyamides, microporous modacrylic copolymers, microporous styrene-acrylic and its copolymers); porous polysulfones, halogenated poly(vinylidene), polychloroethers, acetal polymers, polyesters prepared by esterification of a dicarboxylic acid or anhydride with an alkylene polyol, poly(alkylenesulfides), phenolics, polyesters, asymmetric porous polymers, cross-linked olefin polymers, hydrophilic microporous homopolymers, copolymers or interpolymers having a reduced bulk density, and other similar materials, poly(urethane), crosslinked chain-extended poly(urethane), poly(imides), poly(benzimidazoles), collodion, regenerated proteins, semi-solid cross-linked polyvinylpyrrolidone), monomeric, dimeric, oligomeric or long-chain, copolymers, block polymers, block co-polymers, polymers, PEG, dextran, modified dextran, polyvinyl alcohol, polyvinylpyrollidone, polyacrylates, polymethacrylates, polyanhydrides, polypeptides, albumin, alginates, amino acids, thiols, amines, carboxylic acids, phospholipids, albumin, dextran, gelatin, poly(ethylene glycerol) (PEG), poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, sialic acid, poly(N- acetylglucosamine) (Chitin), Chitosan, poly (3 -hydroxy valerate), poly(D,L-lactide-co-glycolide), poly(l-lactide-co-glycolide), poly(3 -hydroxybutyrate), poly(4-hydroxybutyrate), poly(3- hydroxybutyrate-co-3 -hydroxy valerate), polyorthoester, polyanhydride, poly(glycolic acid), poly(glycolide), poly(L-lactic acid), poly(L-lactide), poly(D,L-lactic acid), poly(D,L-lactide), poly(L-lactide-co-D,L-lactide), poly(caprolactone), poly(L-lactide-co-caprolactone), poly(D,L- lactide-co-caprolactone), poly(glycolide-co-caprolactone), poly(trimethylene carbonate), polyester amide, poly(glycolic acid-co-trimethylene carbonate), co-poly(ether-esters) (e g. PEO / PLA), polyphosphazenes, fibrin, fibrin glue, fibrinogen, cellulose, starch, collagen and hyaluronic acid, elastin and hyaluronic acid, polyurethanes, silicones, polyesters, polyolefins, polyisobutylene and ethylene-alphaolefin copolymers, acrylic polymers and copolymers other than polyacrylates, vinyl halide polymers and copolymers, polyvinyl chloride, polyvinyl ethers, polyvinyl methyl ether, polyvinylidene halides, polyvinylidene chloride, poly (vinylidene fluoride), poly(vinylidene fluoride-co-hexafluoropropylene), polyacrylonitrile, polyvinyl ketones, polyvinyl aromatics, polystyrene, polyvinyl esters, polyvinyl acetate, acrylonitrilestyrene copolymers, ABS resins, polyamides, Nylon 66, poly caprolactam, polycarbonates including tyrosine-based polycarbonates, polyoxymethylenes, polyimides, polyethers, polyurethanes, rayon, rayon-triacetate, cellulose, cellulose acetate, cellulose butyrate, cellulose acetate butyrate, cellophane, cellulose nitrate, cellulose propionate, cellulose ethers, carboxymethyl cellulose, fullerenes, lipids, or any combination thereof.

[0144] In some embodiments, the coating agent comprises at least one molecules selected from gelatin, albumin, dextrose, dextran, a high molecular weight poly(ethylene glycol) or a high molecular weight poly(vinylpyrrolidone), hyaluronic acid, heparin, heparin sulfate, sialic acid, Chitosan, or any combination thereof.

[0145] In some embodiments, the nanoparticles comprise a biodegradable polymer comprises PLGA, poly(D,L-lactide-co-glycolide), poly(D,L-lactide), poly(D,L-lactide-co- lactide), poly(L-lactide), poly(glycolide), poly(L-lactide-co-glycolide), poly(caprolactone), poly(glycolide-co-trimethylene carbonate), poly (3 -hydroxybutyrate), poly (3 -hydroxyb utyrate- co-3 -hydroxy valerate), poly(4-hydroxybutyrate), poly(ester amide), poly(ester-sulfoester amide), poly(orthoester) or poly(anhydride), and a combination thereof.

[0146] In various embodiments, the nanoparticle has an average hydrodynamic diameter of from about 10 nm to about 10,000 nm, from about 130 nm to about 2,500 nm, from about 10 nm to about 1,000 nm, from about 10 nm to about 500 nm, from about 10 nm to about 100 nm, from about 10 nm to about 80 nm, from about 10 nm to about 70 nm, from about 20 nm to about 60 nm, about 50 nm, from about 20 nm to about 50 nm, about 45 nm, of about 30 nm, from about 50 nm to about 150 nm, from about 20 nm to about 80 nm, or about 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, 900 nm, 1,000 nm, 2,000 nm, 2,500 nm, 5,000 nm, or 10,000 nm.

[0147] For example, in various embodiments, the nanoparticle has an average particle size (e g., average hydrodynamic diameter of the nanoparticle) below about 2,500 nm. In some embodiments, the nanoparticle has a particle size (e.g., average hydrodynamic diameter of the nanoparticle) of between about 1 nm to about 2,500 nm. In some embodiments, the nanoparticle has a particle size (e.g., average hydrodynamic diameter of the nanoparticle) of between about 1 nm to about 300 nm. In some embodiments, the nanoparticle has a particle size (e.g., average hydrodynamic diameter of the nanoparticle) of between about 1 nm to about 200 nm.

[0148] In some embodiments, the nanoparticle has a narrow size distribution. In some embodiments, the nanoparticles have a poly dispersity index (PDI) of less than about 0.4, less than about 0.35, less than about 0.3, less than about 0.25, less than about 0.225, less than about 0.2, less than about 0.1. In some embodiments, the nanoparticles have a PDI between about 0.1 and about 0.01, between about 0.09 and about 0.02, between about 0.08 and about 0.03, between about 0.07 and about 0.04, between about 0.06 and about 0.05.

[0149] In various embodiments, the nanoparticle of the present invention is substantially non-toxic.

[0150] In one embodiment, the nanoparticle is a biodegradable nanoparticle.

[0151] In some embodiments, the nanoparticle is any type of nanoparticle, including, but not limited to, liposomes, lipid nanoparticles, organic nanoparticles, inorganic nanoparticles (e.g., metal nanoparticles, such as gold nanoparticles, iron nanoparticles, ZnO nanoparticles, TiCh nanoparticles, etc.), biocompatible nanoparticles, such as biocompatible organic nanoparticles, biocompatible inorganic nanoparticles, etc., polymer nanoparticles, nanoclusters, nanocapsules, core-shell nanocapsules, nanovesicles, micelles, block copolymer micelles, lamaellae shaped particles, polymersomes, dendrimers, emulsions, exosomes, SEDDS, microspheres, micro- structured lipid carriers, nano-structured lipid carriers, and other nano-size particles of various other small fabrications that are known to those of skill in the art.

[0152] For example, in some embodiments, the nanoparticle is a liposome, lipid nanoparticle, nanocapsule, nanocarrier, nanoemulsion, micelle, solid lipid nanoparticle (SLN), nanostructured lipid carrier (NLC), or any combination thereof.

[0153] In one embodiment, the nanoparticle is a liposome. In one embodiment, the nanoparticle is a biodegradable liposome. In some embodiments, the liposome comprises phospholipids, cholesterol, sphingolipids, ceramides, hapten-conjugated lipids, or any combination thereof.

[0154] In one embodiment, the nanoparticle is a lipid nanoparticle. In some embodiments, the lipid nanoparticles comprise phospholipids, cholesterol, PEG-phospholipids, lipid-soluble vitamins, lipid conjugates, or any combination thereof.

[0155] In one aspect, the invention is not limited to any particular cargo or agent for which the nanoparticle of the invention is able to carry or transport. Rather, the invention includes any agent that can be carried by the nanoparticle of the invention. For example, the agents that can be carried by the nanoparticle of the invention include, but are not limited to, therapeutic agents.

[0156] In some embodiments, the agent is adhered to the surface of the nanoparticle of the invention. In some embodiments, the agent is integrated into the structure of the nanoparticle of the invention. In some embodiments, the nanoparticle of the invention encapsulates at least one agent. In some embodiments, the nanoparticle of the invention comprises at least two agents.

[0157] In one aspect of the invention, the agent is a therapeutic agent. In one embodiment, the nanoparticle or composition of the invention encapsulates the therapeutic agent. In one embodiment, the nanoparticle or composition of the invention is bound to the therapeutic agent. In one embodiment, the therapeutic agent is a hydrophobic therapeutic agent. In one embodiment, the therapeutic agent is a hydrophilic therapeutic agent. Examples of such therapeutic agents include, but are not limited to, at least one drugs, proteins, amino acids, peptides, antibodies, antibiotics, small molecules, anti-cancer agents, chemotherapeutic agents, immunomodulatory agents, RNA molecules, siRNA molecules, DNA molecules, gene editing agents, gene-silencing agents, CRISPR-associated agents (e.g., guide RNA molecules, endonucleases, and variants thereof), medical imaging agents, therapeutic moieties, at least one non-therapeutic moieties or a combination to target cancer or atherosclerosis, selected from folic acid, peptides, proteins, aptamers, antibodies, siRNA, poorly water soluble drugs, anti-cancer drugs, antibiotics, analgesics, vaccines, anticonvulsants; anti-diabetic agents, antifungal agents, antineoplastic agents, anti-parkinsonian agents, anti-rheumatic agents, appetite suppressants, biological response modifiers, cardiovascular agents, central nervous system stimulants, contraceptive agents, dietary supplements, vitamins, minerals, lipids, saccharides, metals, amino acids (and precursors), nucleic acids and precursors, contrast agents, diagnostic agents, dopamine receptor agonists, erectile dysfunction agents, fertility agents, gastrointestinal agents, hormones, immunomodulators, anti-hypercalcemia agents, mast cell stabilizers, muscle relaxants, nutritional agents, ophthalmic agents, osteoporosis agents, psychotherapeutic agents, parasympathomimetic agents, parasympatholytic agents, respiratory agents, sedative hypnotic agents, skin and mucous membrane agents, smoking cessation agents, steroids, sympatholytic agents, urinary tract agents, uterine relaxants, vaginal agents, vasodilator, anti-hypertensive, hyperthyroids, anti-hyperthyroids, anti-asthmatics and vertigo agents, or any combinations thereof. For example, in one embodiment, the therapeutic agent is a therapeutic agent for preventing or treating metabolic diseases or disorders.

[0158] In one embodiment, the agent is at least one non-therapeutic moieties. In some embodiments, the nanoparticle or composition of the invention comprises at least one therapeutic moiety, at least one non-therapeutic moiety, or any combination thereof.

[0159] In some embodiments, the nanoparticle or composition further comprises a biocompatible metal. Examples of biocompatible metals include, but are not limited to, copper, copper sulfide, iron oxide, cobalt and noble metals, such as gold and / or silver. One of ordinary skill in the art will be able to select of a suitable type of nanoparticle or composition taking into consideration at least the type of therapy to be performed.

[0160] One of skill in the art would understand that different embodiments of nanoparticles disclosed herein can be used in connection with any aspect of the described invention.

[0161] In various embodiment, nanoparticle or composition increases bioavailability of resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0162] In one aspect of the invention, the nanoparticle or composition induces browning of a white adipose tissue (WAT), beige adipose tissue, or any combination thereof; increases the level or activity of a brown adipose tissue (BAT), beige adipose tissue, or any combination thereof; increases the concentration of cellular cyclic adenosine monophosphate (cAMP), increases the concentration of a an uncoupling protein 1 (UCP1); increases the level or activity of a PR domain-containing 16 (PRDM16); increases the level or activity of a 5' adenosine monophosphate-activated protein kinase (AMPK); increases the level or activity of sirtuin 1 (SIRT1); increases the level or activity of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-la); increase expression of a tight junction protein-related gene (e.g., occluding (OCLN); decreases the level or activity of a fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), or a combination thereof; decreases the level or activity of NF-KB; decreases the level or activity of NF-KB, interferon-y, Interleukin 6 (IL-6), IL-4, IL-1 a, monocyte chemoOattractant protein 1 (MCP-6), tumor necrosis factor alpha (TNF-a), or a combination thereof; decreases the level or activity of a phosphodiesterase (PDE); or any combination thereof.

[0163] In one aspect of the invention, the nanoparticle or composition increases a mitochondrial biogenesis, mitochondrial function, thermogenesis, or a combination thereof; increases gut integrity, gut health, or a combination thereof; increases insulin sensitivity; increases energy expenditure of at least one cell of interest; improves metabolic health; improves gut microbiota profile; improves glucose homeostasis, lipid homeostasis, or a combination thereof; decreases fat mass and body weight; decreases inflammatory response; decrease liver damage; or any combination thereof.

[0164] In one embodiment, the nanoparticle does not change the level or concentration of lean mass.

[0165] In some aspects, the present invention provides a composition comprising at least one nanoparticle of the present invention, resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, at least one lipid, at least one surfactant, and at least one vitamin E. In some embodiments, the composition is oral formulation, an injectable solution, a solution for transdermal delivery, an injectable hydrogel, a surgically implantable hydrogel, and any combination thereof.

[0166] Methods

[0167] In one aspect, the present invention provides a method of delivering resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, to a subject. In some embodiments, the present invention provides a method of delivering resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, to a subject in need thereof. In some embodiments, a subject in need has at least one metabolic disease or disorder.

[0168] In some embodiments, the present invention provides a method of delivering resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, to an adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.). In some embodiments, the present invention provides a method for in vivo delivery of resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, to an adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.).

[0169] In one aspect, the present invention provides a method of increasing the concentration of a cellular cAMP in an adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.). In one aspect, the present invention provides a method of browning an adipose tissue of interest (e.g., WAT, beige adipose tissue, etc.). In one aspect, the present invention provides a method of activating an adipose tissue of interest (e.g., BAT, etc.). In one embodiment, the method increases the level or activity of a 5' adenosine monophosphate- activated protein kinase (AMPK). In one embodiment, the method increases the level or activity of sirtuin 1 (SIRT1). In one embodiment, the method increases the level or activity of peroxisome proliferator-activated receptor gamma coactivator 1 -alpha (PGC-la). In one embodiment, the method decreases the level or activity of a fatty acid synthase (FAS), acetyl- CoA carboxylase (ACC), or a combination thereof. In one embodiment, the method decreases the level or activity of NF-KB. In one embodiment, the method decreases the level or activity of interferon-y, IL-4, IL- la, or a combination thereof. In one embodiment, the method decreases the level or activity of a phosphodiesterase (PDE). In one aspect, the present invention provides a method of reducing a percent body fat composition in a subject in need thereof. In another aspect, the present invention provides a method of inducing weight loss in a subject in need thereof.

[0170] In another aspect, the present invention provides a method of treating a metabolic disease or disorder in a subject in need thereof. In some embodiments, the metabolic disease or disorder is selected from obesity, diabetes, heart disease, stroke, cardiovascular disease, hypertension, idiopathic intracranial hypertension, coronary artery disease, atrial fibrillation, atherosclerosis, venous thromboembolism, obstructive sleep apnea , obesity hypoventilation syndrome, metabolic syndrome, insulin resistance, non-alcoholic fatty liver disease, esophageal adenocarcinoma, gastric cancer, colorectal cancer, hepatocellular carcinoma, cholangiocarcinoma, pancreatic cancer, endometrial carcinoma, ovarian cancer, breast cancer, renal cell carcinoma, multiple myeloma, or any combination thereof. In some embodiments, the metabolic disease or disorder is obesity. Thus, in one aspect, the present invention provides a method of treating obesity in a subject in need thereof.

[0171] In one aspect, the present invention relates to a method of inducing browning of an adipose tissue in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle described above; at least one composition described above; resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof; or any of combination thereof.

[0172] In one aspect, the present invention relates to a method of reducing body fat in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle described above; at least one composition described above; resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof; or any of combination thereof.

[0173] In one aspect, the present invention relates to a method of inducing weight loss in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle described above; at least one composition described above; resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof; or any of combination thereof.

[0174] In one aspect, the present invention relates to a method of treating or preventing a metabolic disease or disorder in a subject in need thereof, wherein the method comprises administering to the subject: at least one nanoparticle described above; at least one composition described above; resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof; or any of combination thereof.

[0175] In one embodiment, the method further comprises a fecal microbiota transplantation. In one embodiment, the fecal microbiota transplantation is a human fecal microbiota transplantation (HFT).

[0176] In one aspect, the present invention relates to a method of optimizing or improving the gut microbiota in a subject in need thereof, wherein the method comprises administering to the subject: a) a fecal microbiota transplantation; and b) at least one nanoparticle described above; at least one composition described above; resveratrol, or derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof; or any of combination thereof.

[0177] In various embodiments, the method comprises administering an effective amount of resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, to a subject in need. In some embodiments, the method comprises subcutaneously injecting an effective amount of resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, into an adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.) of a subject. In various embodiments, the method comprises administering an effective amount of at least one nanoparticle of the present invention to a subject in need. In some embodiments, the nanoparticle is targeted for an adipose tissue of interest (e g., WAT, beige adipose tissue, BAT, etc ). In some embodiments, the method comprises administering an effective amount of at least one composition of the present invention to the subject. In some embodiments, the composition comprises at least one nanoparticle targeted for an adipose tissue of interest (e.g., WAT, beige adipose tissue, BAT, etc.).

[0178] In one embodiment, the method comprises administration of the composition to a subject. In certain embodiments, the method comprises administering a plurality of doses to the subject. In another embodiment, the method comprises administering a single dose of the composition, where the single dose is effective in delivery of the target therapeutic agent.

[0179] Administration of the compositions of the invention in a method of treatment can be achieved in a number of different ways, using methods known in the art. In one embodiment, the method of the invention comprises systemic administration of the subject, including for example enteral or parenteral administration, oral administration, injection, or a combination thereof. In some embodiments, the method comprises intrafat delivery of the composition. In certain embodiments, the method comprises intradermal delivery of the composition. In another embodiment, the method comprises intravenous delivery of the composition. In some embodiments, the method comprises intramuscular delivery of the composition. In one embodiment, the method comprises subcutaneous delivery of the composition. In one embodiment, the method comprises inhalation of the composition. In one embodiment, the method comprises intranasal delivery of the composition. In one embodiment, the method comprises transdermal delivery.

[0180] In some embodiments, administration comprises intravenous, intranasal, intramuscular, subcutaneous, transdermal, or intrafat delivery of resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, and / or quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof nanoparticle and / or composition of the present invention.

[0181] It will be appreciated that the composition of the invention may be administered to a subject either alone, or in conjunction with another agent.

[0182] The pharmaceutical compositions useful for practicing the invention may be administered to deliver a dose of from 0.1 ng / kg / day and 100 mg / kg / day. In one embodiment, the invention envisions administration of a dose which results in a concentration of the compound of the present invention from 10 nM and 10 pM in a mammal.

[0183] Typically, dosages which may be administered in a method of the invention to a mammal, for example a human, range in amount from 0.01 pg to about 50 mg per kilogram of body weight of the mammal, while the precise dosage administered will vary depending upon any number of factors, including but not limited to, the type of mammal and type of disease state being treated, the age of the mammal and the route of administration. In one embodiment, the dosage of the compound will vary from about 0.1 pg to about 100 mg per kilogram of body weight of the mammal. In one embodiment, the dosage of the compound will vary from about 0.1 pg to about 50 mg per kilogram of body weight of the mammal. In one embodiment, the dosage of the compound will vary from about 0.1 pg to about 10 mg per kilogram of body weight of the mammal. In one embodiment, the dosage will vary from about 1 pg to about 1 mg per kilogram of body weight of the mammal.

[0184] The composition may be administered to a mammal as frequently as several times daily, or it may be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as once every several months or even once a year or less. The frequency of the dose will be readily apparent to the skilled artisan and will depend upon any number of factors, such as, but not limited to, the type and severity of the disease being treated, the type and age of the mammal, etc.

[0185] In certain embodiments, administration of a composition of the present invention may be performed by single administration or boosted by multiple administrations. In one embodiment, the invention includes a method comprising administering a combination of compositions described herein. In certain embodiments, the combination has an additive effect, wherein the overall effect of the administering the combination is approximately equal to the sum of the effects of administering each composition. In other embodiments, the combination has a synergistic effect, wherein the overall effect of administering the combination is greater than the sum of the effects of administering each composition.

[0186] In one aspect, the therapeutic compounds or compositions of the invention may be administered prophylactically (i.e., to prevent obesity or a metabolic disease or disorder) or therapeutically (i.e., to treat obesity or a metabolic disease or disorder) to subjects suffering from or at risk of (or susceptible to) developing the obesity or a metabolic disease or disorder. Such subjects may be identified using standard clinical methods.

[0187] In the context of the present invention, prophylactic administration occurs prior to the manifestation of overt clinical symptoms of a metabolic disease or disorder, such that metabolic disease or disorder is prevented or alternatively delayed in its progression. In the context of the field of medicine, the term “prevent” encompasses any activity which reduces the burden of mortality or morbidity from a disease. Prevention can occur at primary, secondary and tertiary prevention levels. While primary prevention avoids the development of a disease, secondary and tertiary levels of prevention encompass activities aimed at preventing the progression of a disease and the emergence of symptoms as well as reducing the negative impact of an already established disease by restoring function and reducing disease-related complications.

[0188] The composition of the invention can be useful in combination with therapeutic, anti-cancer, and / or radiotherapeutic agents. Thus, the present disclosure provides a combination of the present nanoparticle with therapeutic, anti-cancer, and / or radiotherapeutic agents for simultaneous, separate, or sequential administration. The composition of the invention and the other anticancer agent can act additively or synergistically.

[0189] The therapeutic agent, anti-cancer agent, and / or radiation therapy can be administered according to therapeutic protocols well known in the art. It will be apparent to those skilled in the art that the administration of the therapeutic agent, anti-cancer agent, and / or radiation therapy can be varied depending on the disease being treated and the known effects of the anti -cancer agent and / or radiation therapy on that disease. Also, in accordance with the knowledge of the skilled clinician, the therapeutic protocols (e.g., dosage amounts and times of administration) can be varied in view of the observed effects of the administered therapeutic agents (i.e., anti -neoplastic agent or radiation) on the patient, and in view of the observed responses of the disease to the administered therapeutic agents, and observed adverse effects.

[0190] Pharmaceutical Compositions

[0191] The formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or at least one other accessory ingredient, and then, if necessary or desirable, shaping or packaging the product into a desired single- or multi-dose unit.

[0192] Although the description of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for ethical administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the pharmaceutical compositions of the invention is contemplated include, but are not limited to, humans and other primates, mammals including commercially relevant mammals such as non-human primates, cattle, pigs, horses, sheep, cats, and dogs.

[0193] Pharmaceutical compositions that are useful in the methods of the invention may be prepared, packaged, or sold in formulations suitable for ophthalmic, oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, intravenous, intracerebroventricular, intradermal, intramuscular, or another route of administration. Other contemplated formulations include projected nanoparticles, liposomal preparations, resealed erythrocytes containing the active ingredient, and immunogenic-based formulations.

[0194] A pharmaceutical composition of the invention may be prepared, packaged, or sold in bulk, as a single unit dose, or as a plurality of single unit doses. As used herein, a “unit dose” is discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.

[0195] The relative amounts of the active ingredient, the pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical composition of the invention will vary, depending upon the identity, size, and condition of the subject treated and further depending upon the route by which the composition is to be administered. By way of example, the composition may comprise between 0.1% and 100% (w / w) active ingredient.

[0196] In addition to the active ingredient, a pharmaceutical composition of the invention may further comprise at least one additional pharmaceutically active agent.

[0197] Controlled- or sustained-release formulations of a pharmaceutical composition of the invention may be made using conventional technology.

[0198] As used herein, “parenteral administration” of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subject and administration of the pharmaceutical composition through the breach in the tissue. Parenteral administration thus includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like. In particular, parenteral administration is contemplated to include, but is not limited to, intrafat, transdermal delivery, intraocular, intravitreal, subcutaneous, intraperitoneal, intramuscular, intradermal, intrastemal injection, intratumoral, intravenous, intracerebroventricular and kidney dialytic infusion techniques.

[0199] Formulations of a pharmaceutical composition suitable for parenteral administration comprise the active ingredient combined with a pharmaceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further comprise at least one additional ingredient including, but not limited to, suspending, stabilizing, or dispersing agents. In one embodiment of a formulation for parenteral administration, the active ingredient is provided in dry (i.e. powder or granular) form for reconstitution with a suitable vehicle (e.g. sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

[0200] The pharmaceutical compositions may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to the known art, and may comprise, in addition to the active ingredient, additional ingredients such as the dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations may be prepared using a non-toxic parenterally-acceptable diluent or solvent, such as water or 1,3-butane diol, for example. Other acceptable diluents and solvents include, but are not limited to, Ringer’s solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or di-glycerides. Other parentally-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer systems. Compositions for sustained release or implantation may comprise pharmaceutically acceptable polymeric or hydrophobic materials such as an emulsion, an ion exchange resin, a sparingly soluble polymer, or a sparingly soluble salt.

[0201] A pharmaceutical composition of the invention may be prepared, packaged, or sold in a formulation suitable for pulmonary administration via the buccal cavity. Such a formulation may comprise dry particles which comprise the active ingredient and which have a diameter in the range from about 0.5 to about 7 nanometers, or from about 1 to about 6 nanometers. Such compositions are conveniently in the form of dry powders for administration using a device comprising a dry powder reservoir to which a stream of propellant may be directed to disperse the powder or using a self-propelling solvent / powder-dispensing container such as a device comprising the active ingredient dissolved or suspended in a low-boiling propellant in a sealed container. In one embodiment, such powders comprise particles wherein at least 98% of the particles by weight have a diameter greater than 0.5 nanometers and at least 95% of the particles by number have a diameter less than 7 nanometers. In one embodiment, at least 95% of the particles by weight have a diameter greater than 1 nanometer and at least 90% of the particles by number have a diameter less than 6 nanometers. In one embodiment, dry powder compositions include a solid fine powder diluent such as sugar and are conveniently provided in a unit dose form.

[0202] Low boiling propellants generally include liquid propellants having a boiling point of below 65°F at atmospheric pressure. Generally the propellant may constitute 50 to 99.9% (w / w) of the composition, and the active ingredient may constitute 0.1 to 20% (w / w) of the composition. The propellant may further comprise additional ingredients such as a liquid non-ionic or solid anionic surfactant or a solid diluent (in some aspects, having a particle size of the same order as particles comprising the active ingredient).

[0203] Formulations of a pharmaceutical composition suitable for parenteral administration comprise the active ingredient combined with a pharmaceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations may further comprise at least one additional ingredient including, but not limited to, suspending, stabilizing, or dispersing agents. In one embodiment of a formulation for parenteral administration, the active ingredient is provided in dry (i.e., powder or granular) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

[0204] The pharmaceutical compositions may be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution may be formulated according to the known art, and may comprise, in addition to the active ingredient, additional ingredients such as the dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations may be prepared using a non-toxic parent erally-acceptable diluent or solvent, such as water or 1,3-butane diol, for example. Other acceptable diluents and solvents include, but are not limited to, Ringer’s solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or di-glycerides. Other parentally-administrable formulations that are useful include those that comprise the active ingredient in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer system. Compositions for sustained release or implantation may comprise pharmaceutically acceptable polymeric or hydrophobic materials such as an emulsion, an ion exchange resin, a sparingly soluble polymer, or a sparingly soluble salt.

[0205] As used herein, “additional ingredients” include, but are not limited to, at least one of the following: excipients; surface active agents; dispersing agents; inert diluents; granulating and disintegrating agents; binding agents; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives; physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents, demulcents; buffers; salts; thickening agents; fillers; emulsifying agents; antioxidants; antibiotics; antifungal agents; stabilizing agents; and pharmaceutically acceptable polymeric or hydrophobic materials. Other “additional ingredients” which may be included in the pharmaceutical compositions of the invention are known in the art and described, for example in Remington’s Pharmaceutical Sciences (1985, Genaro, ed., Mack Publishing Co., Easton, PA), which is incorporated herein by reference.

[0206] EMBODIMENTS

[0207] Embodiment l is a nanoparticle comprising: a) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, wherein resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, is present in a concentration range of about 0.1 wt% to about 90 wt%; b) quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, wherein quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, is present in a concentration range of about 0.1 wt% to about 90 wt%; c) at least one lipid, wherein the lipid is present in a concentration range of about 5 wt% to about 99 wt%; d) at least one surfactant, wherein the surfactant is present in a concentration range of about 10 wt% to about 75 wt%; and e) at least one vitamin E, wherein the vitamin E is present in a concentration range of about 5 wt% to about 85 wt%.

[0208] Embodiment 2 is the nanoparticle of embodiment 1, wherein the at least one lipid is selected from the group consisting of a fatty acid, wax, sterol, lipid-soluble vitamin, vitamin A, vitamin D, vitamin E, vitamin K, monoglyceride, diglyceride, triglyceride, phospholipid, phosphatidylcholine, L-a-phosphatidylcholine (PC), soy PC, egg PC, phosphatidic acid (PA), phosphatidylethanolamine (PE), lecithin, phosphatidyl serine (PS), phosphoinositide, phosphatidylinositol (PI), phosphatidylinositol phosphate (PIP), phosphatidylinositol bisphosphate (PIP2), phosphatidylinositol trisphosphate (PIP3), phosphosphingolipid, sphingolipid, ceramide phosphorylcholine (SPH), ceramide phosphorylethanolamine (Cer-PE), ceramide phosphoryl lipid, and any combination thereof.

[0209] Embodiment 3 is the nanoparticle of embodiment 1 or 2, wherein the at least one lipid is phosphatidylcholine.

[0210] Embodiment 4 is the nanoparticle of any one of embodiments 1-3, wherein the at least one surfactant is selected from the group consisting of a polyethylene glycol (PEG), functionalized PEG, polyethylene glycol (15)-hydroxy stearate , polyethylene glycol (15)- hydroxy stearate containing PEG, polysorbate 80, phospholipids, PEG21 -stearate, PEG40- stearate, PEGlOO-stearate, PEG(10-1000)-fatty acid, mustard, lecithin, soy lecithin, egg lecithin, monoglycerides, diglycerides, polysorbates, carrageenan, guar gum, canola oil, polysorbates, sodium dodecyl sulfate, lauryl dimethyl amine oxide, cetyltrimethylammonium bromide (CTAB), polyethoxylated alcohols, polyoxyethylene sorbitan, octoxynol, N, N- dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide (HTAB), polyoxyl 10 lauryl ether, bile salts, sodium deoxycholate, sodium cholate, polyoxyl castor oil, nonylphenol ethoxylate, cyclodextrins, methylbenzethonium chloride, and any combination thereof.

[0211] Embodiment 5 is the nanoparticle of any one of embodiments 1-4, wherein the at least one surfactant is polyethylene glycol (15)-hydroxystearate , polyethylene glycol (15)- hydroxystearate containing PEG.

[0212] Embodiment 6 is the nanoparticle of any one of embodiments 1-5, wherein the at least one vitamin E is selected from the group consisting of a-tocopherol, a-tocopherol acetate (aTA), a-tocopherol nicotinate, D-a-tocopheryl polyethylene glycol 1000 succinate, P-tocopherol, 0- tocopherol acetate, P-tocopherol nicotinate, y-tocopherol, ' / -tocopherol acetate, y-tocopherol nicotinate, 8-tocopherol, 8-tocopherol acetate, 8-tocopherol nicotinate, a-tocotrienol, a- tocotrienol acetate, a-tocotrienol nicotinate, P-tocotrienol, P-tocotrienol acetate, P-tocotrienol nicotinate, y-tocotrienol, y-tocotrienol acetate, y-tocotrienol nicotinate, 5-tocotrienol, 8- tocotrienol acetate, 8-tocotrienol nicotinate, and any combination thereof.

[0213] Embodiment 7 is the nanoparticle of any one of embodiments 1-6, wherein the at least one vitamin E is aTA.

[0214] Embodiment 8 is the nanoparticle of any one of embodiments 1-7, wherein the nanoparticle further comprises at least one targeting agent.

[0215] Embodiment 9 is the nanoparticle of embodiment 8, wherein the at least one targeting agent targets an adipose tissue.

[0216] Embodiment 10 is the nanoparticle of embodiment 9, wherein the adipose tissue is a white adipose tissue (WAT), beige adipose tissue, brown adipose tissue (BAT), or any combination thereof.

[0217] Embodiment 11 is the nanoparticle or 8-10, wherein the targeting agent is selected from the group consisting of proteins, peptides, antibodies, lipids, polysaccharides, nucleic acids, aptamers, and small molecules.

[0218] Embodiment 12 is the nanoparticle of any one of embodiments 1-11, wherein the composition further comprises at least one phytochemical or bioactive compound selected from the group consisting of curcumin, theaflavins, thearubigins, epigallocatechin gallate (EGCG), (+)-catechin, (-)-epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin, (+)-gallocatechin, isorhamnetin, kaempferol, myricetin, apigenin, luteolin, baicalein, chrysin, forskolin, chlorophyll a, chlorophyll b, eriodictyol, hesperetin, naringenin, taxifolin, catechins, luteolin, cyanidin, genistein, daidzein, genistein, glycitein, biochanin A, formononetin, cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, proanthocyanidins, a-carotene, P-carotene, P- cryptoxanthin, lutein, zeaxanthin, lycopene, and any combination thereof.

[0219] Embodiment 13 is the nanoparticle of any one of embodiments 1-12, wherein the nanoparticle further comprises at least one coating agent.

[0220] Embodiment 14 is the nanoparticle of any one of embodiments 1-13, wherein the nanoparticle is a biodegradable nanoparticle.

[0221] Embodiment 15 is the nanoparticle of any one of embodiments 1-14, wherein the nanoparticle increases bioavailability of resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0222] Embodiment 16 is the nanoparticle of any one of embodiments 1-15, wherein the nanoparticle: a) induces browning of a white adipose tissue (WAT), beige adipose tissue, or any combination thereof; b) increases the level or activity of a brown adipose tissue (BAT), beige adipose tissue, or any combination thereof; c) increases the concentration of cellular cyclic adenosine monophosphate (cAMP); d) increases the level or activity of uncoupling protein 1 (UCP1) or PR domain containing 16 (PRDM16); e) increases the level or activity of a 5' adenosine monophosphate-activated protein kinase (AMPK); f) increases the level or activity of sirtuin 1 (SIRT1); g) increases the level or activity of tight junction protein-related genes; h) increases the level or activity of occluding (OCLN); i) increases the level or activity of peroxisome proliferator-activated receptor gamma coactivator 1 -alpha (PGC-la); j) decreases the level or activity of a fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), or a combination thereof; k) decreases the level or activity of NF-KB, tumor necrosis factor alpha (TNF-a), interferon-y, Interleukin 6 (IL-6), IL-4, IL- la, monocyte chemoattractant protein 1 (MCP1), or a combination thereof; l) decreases the level or activity of a phosphodiesterase (PDE); or any combination thereof.

[0223] Embodiment 17 is the nanoparticle of any one of embodiments 1-16, wherein the nanoparticle: a) increases a mitochondrial biogenesis, mitochondrial function, thermogenesis, or a combination thereof; b) increases gut integrity, gut health, or a combination thereof; c) increases insulin sensitivity; d) increases energy expenditure of at least one cell of interest; e) improves metabolic health; f) improves gut microbiota profile; g) improves glucose homeostasis, lipid homeostasis, or a combination thereof; h) decreases fat mass and body weight; i) decreases inflammatory response; j) decrease liver damage; or any combination thereof.

[0224] Embodiment 18 is the nanoparticle of any one of embodiments 1-17, wherein the nanoparticle does not change the level or concentration of lean mass.

[0225] Embodiment 19 is a composition comprising at least one nanoparticle of embodiments 1- 18.

[0226] Embodiment 20 is the composition of embodiment 19, wherein the composition is selected from the group consisting of an oral formulation, an injectable solution, a solution for transdermal delivery, an injectable hydrogel, a surgically implantable hydrogel, and any combination thereof.

[0227] Embodiment 21 is the method of increasing the concentration of a cellular cyclic adenosine monophosphate (cAMP) in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of any one of embodiments 1-18; b) at least one composition of embodiment 19 or 20; or c) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0228] Embodiment 22 is the method of embodiment 21, wherein the method increases the concentration of a cellular cyclic adenosine monophosphate (cAMP) in an adipose tissue.

[0229] Embodiment 23 is a method of selectively delivering resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof to an adipose tissue, wherein the method comprises administering: a) at least one nanoparticle of any one of embodiments 1-18; b) at least one composition of embodiment 19 or 20; or c) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0230] Embodiment 24 is a method of inducing browning of an adipose tissue in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of any one of embodiments 1-18; b) at least one composition of embodiment 19 or 20; or c) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0231] Embodiment 25 is the method of any one of embodiments 22-24, wherein the adipose tissues is a white adipose tissue (WAT), beige adipose tissue, brown adipose tissue (BAT), or any combination thereof.

[0232] Embodiment 26 is a method of reducing body fat in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of any one of embodiments 1-18; b) at least one composition of embodiment 19 or 20; or c) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0233] Embodiment 27 is a method of inducing weight loss in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of any one of embodiments 1-18; b) at least one composition of embodiment 19 or 20; or c) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0234] Embodiment 28 is a method of treating or preventing a metabolic disease or disorder in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of any one of embodiments 1-18; b) at least one composition of embodiment 19 or 20; or c) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

[0235] Embodiment 29 is the method of embodiment 28, wherein the metabolic disease or disorder is selected from the group consisting of obesity, obesity-related disease or disorder, diabetes, fatty liver disease, non-alcoholic fatty liver disease, cardiovascular disease, heart disease, stroke, hypertension, idiopathic intracranial hypertension, coronary artery disease, atrial fibrillation, atherosclerosis, venous thromboembolism, obstructive sleep apnea , obesity hypoventilation syndrome, metabolic syndrome, insulin resistance, esophageal adenocarcinoma, gastric cancer, colorectal cancer, hepatocellular carcinoma, cholangiocarcinoma, pancreatic cancer, endometrial carcinoma, ovarian cancer, breast cancer, renal cell carcinoma, multiple myeloma, and any combination thereof.

[0236] Embodiment 30 is the method of any one of embodiments 21-29, wherein the composition is administered via oral administration, injection, or a combination thereof.

[0237] Embodiment 31 is the method of any one of embodiments 21-30, wherein the method further comprises a fecal microbiota transplantation.

[0238] Embodiment 32 is the method of embodiment 31, wherein the fecal microbiota transplantation is a human fecal microbiota transplantation (HFT).

[0239] EXPERIMENTAL EXAMPLES

[0240] The invention is now described with reference to the following Examples. These Examples are provided for the purpose of illustration only, and the invention is not limited to these Examples, but rather encompasses all variations that are evident as a result of the teachings provided herein.

[0241] Example 1 : Nanoencapsulation of Resveratrol and Quercetin

[0242] Obesity prevalence and magnitude are increasing dramatically in the United States and worldwide. More than two-thirds of U.S adults are either overweight or obese. Obesity is characterized by an increase in fat mass. It is strongly and positively associated with metabolic syndrome, which is characterized by abdominal obesity, dyslipidemia and insulin resistance. Two different adipose tissues are found in mammals: white adipose tissue (WAT) is the major site for energy storage; brown adipose tissue (BAT) plays an important role in thermogenic energy expenditure. The expanded WAT in obesity is well-recognized as an active endocrine and inflammatory organ, typically associated with infiltration of pro-inflammatory immune cells, and excess release of pro-inflammatory cytokine / chemokines, adipokines, and free fatty acids into circulation, all of which negatively impact whole body metabolism. If the condition persists, type II diabetes, cardiovascular disease, hypertension and certain types of cancer can develop. In addition to its function in thermogenic effects, BAT is also considered to be a “metabolic sink” for free fatty acids, glucose and other metabolites. Although BAT is a therapeutic target for treating obesity, many adults do not possess active BAT. Even some adults have BAT, their BAT is decreased or even disappeared with aging. Fortunately, beige / brown- like adipocytes, if formed in WAT, also have thermogenic functions similar to the classic brown adipocytes in BAT. Uncoupling protein 1 (UCP-1), predominantly expressed in brown and beige adipocytes, serves as a marker for these cells. UCP-1 found in the inner mitochondrial membrane of brown and beige adipocytes can dissipate the proton electrochemical gradient generated from oxidative phosphorylation in the form of heat (Hales C. M. et al., Prevalence of Obesity and Severe Obesity Among Adults: United States, 2017-2018. NCHS Data Brief. 2020: 1-8; Ogden C. L. et al., Jama., 2020, 324: 1208-1210; Kalupahana N. S. et al., Adv Nutr., 2011, 2:304-16; Xue R. et al., Am. J. Physiol. Endocrinol. Metab., 2014, 306:E363-72; Gesta S. et al., Cell., 2007, 131 :242-56; Townsend K. L. et al., Trends Endocrinol Metab., 2014, 25: 168-77; Rasoulini N. et al., J Clin Endocrinol Metab. 2008, 93:S64-73; Goktas Z. et al., Mediators Inflamm., 2013:861496; Goktas Z. et al., Front Endocrinol (Lausanne). 2013, 4:69; Kajimura S. et al., Cell Metab. 2015, 22:546-59; Porter C. et al., Cell metabolism. 2016, 24:246-255; Virtanen K. A. et al., New England Journal of Medicine. 2009, 360: 1518-1525; Rogers N. H., Annals of medicine. 2015, 47: 142-149; Sidossis L. et al., The Journal of clinical investigation. 2015, 125:478-486; Min S. Y. et al., Nature medicine. 2016, 22:312; Tseng Y. H. et al., Nat Rev Drug Discov. 2010, 9:465-82; Virtanen K. A. et al., N Engl J Med. 2009, 360: 1518-25; Cannon B. et al., Physiol Rev. 2004, 84:277-359).

[0243] The stromal fractions of WAT and BAT contain adipose stromal stem cells (ASCs), which can be differentiated into brown adipocytes and white adipocytes, respectively. The ASC in WAT can also be differentiated into beige adipocytes. The differentiation of ASC in WAT is regulated by some transcriptional factors including CCAAT / enhancer-binding proteins (C / EBPs) and peroxisome-proliferator-activated receptor y (PPARy). Brown adipocyte differentiation is regulated by PPARy, PPARy co-activator la (PGCl ) and transcription factor PR-domain-containing 16 (PRDM16) can specify a brown adipogenic program. The NAD- dependent deacetylase sirtuin 1 (SIRT1) can deacetylate and further activate PPARy, indirectly promoting browning of WAT. This modification is essential for enhancing PPARy-binding activity and recruiting PRDM16 to PPARy, which can specify brown adipogenic lineage transcriptional activity and initiate browning of WAT. Beige adipocytes can also be formed from mature white adipocytes via trans-differentiation. Increasing the amount and / or activity of beige adipocytes offers perspective on the potential preventive and therapeutic applications on obesity and its related metabolic disorders / diseases (Tang W. et al., Science. 2008, 322:583-6; Tran K. V. et al., Cell Metab. 2012, 15:222-9; Tontonoz P. et al., Cell. 1994, 79:1147-56; Wu Z. et al., Curr Opin Cell Biol. 1999, 11 :689-94; Puidserver P. et al., Cell. 1998, 92:829-39; Kajimura S. et al., Nature. 2009, 460: 1154-8; Seale P. et al., Cell Metab. 2007, 6:38-54; Qiang L. et al., Cell. 2012, 150:620-32; Rosenwald M. et al., Adipocyte. 2014, 3:4-9; Vargas-Castillo A. et al., Arch. Med. Res. 2017, 48:401-413; Tseng Y. H. et al., Nat Rev Drug Discov. 2012, 9:465-82).

[0244] Emerging evidence suggests a causal link between microbial dysbiosis and obesity. For example, using a bidirectional Mendelian randomization approach, data from 952 individuals in the LifeLines-DEEP cohort showed that the short-chain fatty acid (SCFA) butyrate generated by a shift toward increased butyrate-producing gut bacteria, such as Eubacterium rectale and Roseburia intestinalis, improved insulin response. Long-term and short-term antibiotic use has also shown causation: changing the gut microbiome profde by decreasing its diversity, if persists, it increases obesity and type 2 diabetes. However, while long-term use increases these conditions, short-term use can change luminal SCFAs, particularly butyrate, and shift colonic metabolism toward glucose utilization. A recent review covers additional support from mouse and human studies showing that critical shifts from proteolytic to saccharolytic fermentation could decrease or prevent metabolic diseases by decreasing harmful ammonia, phenols and branched-chain fatty acids. Further correlational studies have demonstrated that obese subjects compared to lean subjects have different gut microbiota composition, supporting the universality of the gut-profde connection. These data are consistent, showing obesity is associated with changed Firmicutes / Bacteroidetes ratios, increased Lactobacillus reuteri, bacterial family Christensenel laceae, Clostridium, Oscillibacter, Hydrogenoanaerobacterium, and decreased Lactobacillus plantanim in human and research animal guts. Finally, high fat diets have been shown to ultimately drive these changes in the gut microbiota (Sanna S. et al., Nat Genet. 2019, 51 :600-605; Zarrinpar A. et al., Nat Commun. 2018, 9:2872; Canfora E. E. et al., Nat Rev Endocrinol. 2019, 15:261-273; Lynch S. V. et al., N. Engl. J. Med. 2016, 375:2369- 2379; Turnbaugh P. J. et al., Nature. 2006, 444: 1027-31; Peters B. A. et al., Sci. Rep. 2018, 8:9749; Liu R. et al., Nat Med. 2017, 23:859-868; Maruvada P. et al., Cell Host Microbe. 2017, 22:589-599; Ley R. E. et al., Proc. Natl. Acad. Sci. USA. 2005, 102:11070-5; Duncan S. H. et al., Int. J. Obes. (Lond). 2008, 32: 1720-4; Million M. et al., Int. J. Obes. (Lond). 2012, 36:817- 25; Vandeputte D. et al., Nature. 2017, 551 :507-511; Bondia-Pons I. et al., FASEB J. 2014, 28:4169-79; Cani P. D. et al., Diabetes. 2008, 57: 1470-81).

[0245] Many bioactive compounds show a promise in combating obesity and its related metabolic disorders, but their low levels of aqueous solubility, stability, and bioavailability in the body make administering them in physiological doses unrealistic. This is particularly true to / ra / 7.s-resveratrol (R) and quercetin (Q), which demonstrated promise in preventing and / or treating obesity and its associated metabolic dysfunction in some clinical trials.

[0246] Resveratrol (3,5,4'-trihydroxy- / ran5-stilbene, R) is a natural polyphenolic compound, abundant in red grape skin, in the leaf epidermis of the grape vine, peanuts, and mulberry. Many cell culture studies have demonstrated that R at concentrations between 10 to 100 pM exhibits anti-obesity activities by inducing adipocyte apoptosis, decreasing adipocyte differentiation and proliferation, causing cell cycle arrest, lowering lipogenesis, enhancing fatty acid beta (P)-oxidation, mitochondria biogenesis, and activities, as well as inhibiting inflammatory response. If enough R is delivered, it can induce browning of subcutaneous WAT of obese mice. When mice are given a high fat diet containing 0.04% to 0.4% (w / w) of R supplement, R decreases visceral adipogenesis and inflammatory response, enhances insulin sensitivity, lowers liver fat content, increases mitochondrial biogenesis, mimics diet restriction response and improves survival and health of those mice on a high-fat diet. With orally administered R supplement (80 and 480 mg / day for the 1stand 2ndyear, respectively), adipocyte size decreased and insulin sensitivity improved in visceral WAT of rhesus monkeys on a high- fat, high-sugar diet for 2 years. R can directly inhibit cAMP phosphodiesterases (PDEs), which degrade cAMP. Increased cellular cAMP concentrations can activate AMPK, which further activates SIRT1 and PGC-la, resulting in increased mitochondrial biogenesis and function and decreased inflammation. R has strong anti-inflammatory effects in studies using in vitro cellbased experiments, in vivo animal models, and human clinical trials. SIRT1 deacetylase activity can deacetylate forkhead box protein 01 (F0X01) and NF-KB, which are key mediators in regulating inflammation and other related pathways (Fremont L. et al., Life Sci. 2000, 66:663- 73; Bums J. et al., J. Agric. Food Chem. 2002, 50:3337-40; Hurst W. J. et al., J, Agric. Food Chem. 2008, 56:8374-8; Wang S. et al., J. Nutr. Biochem. 2014, 25: 1-18; Dong W. et al., Int. J. Mol. Med. 2014, 33:1161-8; Prasad K. et al., Int. J. Angiol. 21:7-18; Zu Y. et al., J. Control Release. 2021, 333:339-351; Baur J. A. et al., Nature. 2006, 444:337-42; Kim S. et al., Biochem. Pharmacol. 2011, 81 : 1343-51; Jimenez-Gomez Y. et al., Cell. Metab. 2013, 18:533-45; Park S. J. et al., Cell. 2012, 148:421-33; Chung J. H. et al., Aging. 2012, 4: 144-5; Pacholec M. et al., The Journal of Biological Chemistry. 2010, 285:8340-51; Beher D. et al., Chemical Biology & Drug Design. 2009, 74:619-24; Tennen R. I. et al., Cell. 2012, 148:387-9; Poulsen M. M. et al., Biochim. Biophys. Acta. 2015, 1852: 1124-36).

[0247] Human studies indicate that R can maintain metabolic health, but evidence is inconclusive regarding its effectiveness for obesity prevention or treatment. The major problems with oral administration of R are its low aqueous solubility and bioavailability in humans. The solubility of R in water and physiological fluid is very low, about 0.03 mg / mL; its solubility is high in ethanol (50 mg / mL). The oral absorption of R is about 75% in humans, mainly via transepithelial diffusion. But R’s bioavailability is less than 1%, because majority of R is extensively metabolized by various transformations including glucuronidation, sulfation, methylation, and oxidative degradation. After oral administration, only 1-5% of unmetabolized R is found in the blood, 25% is eliminated in the feces without absorption, and >70% is extensively metabolized in the liver, intestinal tract and gut microbiota. When orally administering about 0.3 mg / kg body weight of R to healthy adult males, blood peak concentrations of R appear at 0.5 h, and the peak plasma unmetabolized R concentration is less than 1 pM. Even when giving human subjects high doses of R (single doses of 0.5, 1, 2.5, or 5 g) via oral administration, the plasma unmetabolized R peaked between 0.8 and 1.5 h at concentrations ranging from 0.3 pM to 2.3 pM. Most of cell studies used R concentrations much higher than its physiological concentrations, which brings questions whether these in vitro data can be used to elucidate the mechanisms in the human body, and how to increase unmetabolized R (active form) concentrations in the body. The peak plasma concentrations of R conjugates were two to forty times higher than those of unmetabolized R. The major R metabolites in the body are glucuronides and sulfate conjugates (inactive forms) that have less bioactivities than unmetabolized R. During absorption in the small intestine, more than 90% of R became glucuronide and sulfate conjugates. These conjugates have less absorption rate than unmetabolized R, because they can be easily effluxed back to the small intestine lumen via the ATP-binding cassette (ABC) family on the enterocytes. The unabsorbed R and its conjugates move to the colon, where gut microbiota further metabolize them. Gut Slackia equolifaciens and Adlercreutzia equolifaciens can reduce R, its glucuronide and sulfate conjugates, and produce dihydroresveratrol, dihydroresveratrol glucuronide and sulfate conjugates, respectively. These polar metabolites can enter the blood stream via enteric absorption, and eventually excreted into the urine. Gut bacterial P-D-glucuronidase and sulfatase can also cleave and remove glucuronide and sulfate and yield a small amount of aglycones. Gut microbiota also produce other metabolites including lunularin and 3,4-dihydroxytrans-stilbene. But the metabolism of these metabolites has not yet been investigated. Absorbed R and its metabolites are further transformed via glucuronidation, sulfation, and methylation in the liver. The liver can excrete about 10% of R metabolites back to the intestinal lumen, presumably via biliary secretion. Enterohepatic recirculation occurs consistently. R-3-O-P-glucuronide, R-3-sulfate, R-4'-O-P-glucuronide are the most abundant metabolites in the blood, urine, and tissues. Majority of these polar metabolites are excreted into the urine by kidneys. A trace amount of R metabolites is accumulated in the tissues / organs. After intragastric administration of R (5.9 mg / kg body weight) to pigs for 6 h, 12 major organs / tissues including the liver, brain, heart, spleen, kidneys, adipose tissue were collected, R and its metabolites were determined in these organs / tissues using HPLC- MS / MS. Up to 0.5% of administered dose were found in these organs / tissues. Both unmetabolized R and R-3-O-glucuronide were detected in subcutaneous and mesenteric WAT, but the unmetabolized R is extremely low, less than 0.01 % of administered R dose. Hence, there is a critical need to increase R aqueous solubility and bioavailability, decrease R metabolism, prolong its circulation time, and enhance R delivery to adipose tissue (Barger J. L., Ann. N. Y. Acad. Sci. 2013, 1290: 122-9; Poulsen M. M. et al., Ann. N. Y. Acad. Sci. 2013, 1290:74-82; Bonechi C. et al., PLoS One. 2012, 7:e41438; Walle T., Ann. N. Y. Acad. Sci. 2011, 1215:9-15; Brill S. S. et al., J. Pharm. Pharmacol. 2006, 58:469-79; Goldberg D. M. et al., Clin. Biochem. 2003, 36:79-87; Boocock D. J. et al., Cancer Epidemiol. Biomarkers Prev. 2007, 16:1246-52; Wenzel E. et al., Mol. Nutr. Food Res. 2005, 49:472-81; Kuhnle G. et al., Biochem. Biophys. Res. Commun. 2000, 272:212-7; Colom H. et al., Pharm. Res. 2011, 28: 1606-21; Wenzel E. et al., Mol. Nutr. Food Res. 2005, 49:482-94; Bird J. K. et al., Adv. Nutr. 2017, 8:839-849; Bode L. M. et al., Am. J. Clin. Nutr. 2013, 97:295-309; Yu C. et al., Pharm. Res. 2002, 19: 1907-14; Azorin-Ortuno M. et al., Mol. Nutr. Food Res. 2011, 55: 1154-68).

[0248] Quercetin (3, 3’, 4’, 5’-7-pentahydroxy flavone, Q) is abundant in onions, apples, red grapes, tomato, broccoli and leafy green vegetables. Q protects against diet induced obesity in high fat diet-induced obese mice by increasing energy expenditure, lowering weight gain, improving blood lipid profile and metabolic condition, and decreasing inflammation. Q can inhibit lipogenesis via down-regulating expression of fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC), increasing energy expenditure via up-regulating expression of CPT-la, PGC-la, and UCP-1, and decrease circulating inflammatory biomarkers including interferon-y, IL-4, and IL-la via suppressing NF-KB activation. A high fat diet supplemented with 0.8%- 1% of Q increased energy expenditure via induce browning of WAT and activating BAT in C57BL / 6J mice. Q exhibits these anti-obesity activities via the p38 mitogen-activated protein kinase (MAPK) and AMPK / SIRT1 signaling pathways. However, the evidence of human studies is inconclusive regarding its effectiveness for combating obesity. Major problems, as with R, are its low solubility and bioavailability in humans or research animals. Q is a poorly-water soluble compound; its solubility in water and physiological fluid is less than 0.003 g / L at 25°C. Additionally, Q has a low level of bioavailability. After oral administration of 1 g / day for 28 days, the plasma Q concentrations in human subjects only reached 1.5 pM. Moreover, Q is extensively metabolized by various transformations including methylation, glucuronidation, sulfation and ring fission in vivo. Q aglycone is absorbed in both stomach and small intestine, but glycosylated forms of Q cannot be absorbed. Beta-glucosidase on the brush border and in the gut microbiota can convert them into aglycone via deglycosylation. Q is metabolized via three phases. Phase I metabolism of Q using in cytochrome P450 enzymes has not been reported. Phase II conjugation of Q at the small intestine produces glucuronidated, sulfated and methylated metabolites. Phase III efflux of phase II Q conjugates occurs at the small intestine. Phase II Q conjugates are also secreted from enterocytes into the vein or lymph, and transported to the liver. Phase II conjugation enzymes in the hepatocytes further metabolize Q and its metabolites. Q metabolites produced in the liver either enter the circulation system, or secrete into the bill for enterohepatic recirculation. The unabsorbed Q and its metabolites move to the colon, in where gut microbiota further metabolize them. The primary Q metabolites are 2-(3,4- dihydroxyphenyl)acetic acid, phloroglucinol, 3,4-dihydroxybenzaldehyde, acetate, butyrate and 3,4-dihydroxytoluene in the colon. Q has a low bioavailability. Following oral administration of Q aglycone at dose 100-1,000 mg, the peak plasma Q concentrations was dose-dependently increased from 0.1 to 2 pM. Increasing Q solubility can enhance its bioavailability. In summary, Q has a poor solubility, and extensive metabolism, and Q metabolites have less anti-obesity bioactivities than unmetabolized Q. Circulating Q cannot target to adipose tissue. These limitations result in Q’s inefficiency in fighting obesity in humans. Hence, there is a critical need to increase Q’s solubility, bioavailability, and delivery to adipose tissue (Manach C. et al., Am. J. Clin. Nutr. 2005, 81 :230S-242S; Williamson G. et al., Am. J. Clin. Nutr. 2005, 81 :243S-255S; Stewart L. K. et al., Metabolism. 2008, 57:S39-46; Jung C. H. et al., Phytother. Res. 2013, 27: 139-43; Rivera L. et al., Obesity (Silver Spring). 2008, 16:2081-7; Chen S. et al., Mediators Inflamm. 2016, 2016:9340637; Ahn J. et al., Biochem. Biophys. Res. Commun. 2008, 373:545- 9; Silvester A. J. et al., J. Nutr. Biochem. 2019, 64: 1-12; Pei Y. et al., J. Nutr. Biochem. 2021, 88: 108532; Zhu M. et al., Planta. Med. 2000, 66:444-7; Sun M. et al., Colloids. Surf. B. Biointerfaces. 2014, 113: 15-24; Smith A. J. et al., Mol. Pharm. 2011, 8:1867-76; Kumari A. et al., PLoS One. 2012, 7:e41230; Guo Y. et al., J. Nutr. Biochem. 2015, 26:201-10; Conquer J. A. et al., J. Nutr. 1998, 128:593-7; Boots A. W. et al., Eur. J. Pharmacol. 2008, 585:325-37; Walle T. et al., J. Nutr. 2001, 131 :2648-52; Murakami A. et al., Cancer Lett. 2008, 269:315-25; Russo G. L., Biochem. Pharmacol. 2007, 74:533-44).

[0249] Biodegradable and biocompatible nanoparticles (NPs) smaller than 100 nm are extravasated effectively into tissues and cleared much slower than large carriers by the reticuloendothelial system in the liver and spleen. Recently, nanostructured lipid carriers have received considerable attention because of their small size, stability, biocompatibility and biodegradability, low cytotoxicity, and easily scaled-up synthesis processes. They are made from lipids including mono-, di- or triglycerides, fatty acids, surfactant(s), and water, and their structure is composed of a hydrophilic shell and a hydrophobic lipid core. Hydrophobic drugs and compounds, such as R and Q, can be easily encapsulated into the lipid core. It is well known that mono-, di- or tri-glycerides and fatty acids are high energy molecules, and increased consumption of these lipids are positively correlated with high risks of obesity, cardiovascular disease and diabetes. These lipids have successfully been replaced with a-tocopherol acetate (Sigma T3001, melting temperature 25°C) and (±)-a-tocopherol nicotinate (Sigma T 5134, melting temperature 38°C). These two Vitamin E derivatives have different structures that can lead to high loading capacity of the hydrophobic core. Additionally, a-tocopherol acetate and (±)-a-tocopherol nicotinate have many beneficial effects including antioxidant property that further protect R and Q from degradation. R and Q-loaded nanoparticles ( RQ-NPs), are composed of R, Q, phosphatidylcholine, a-tocopherol acetate, (±)-a-tocopherol nicotinate, and Kolliphor® HS15. All NP components are generally recognized as safe (GRAS) by the U.S. Food and Drug Administration (FDA). NPs with different melting temperatures can be prepared by changing amounts of a-tocopherol acetate and (±)-a-tocopherol nicotinate. For example, they can be solid at room temperature or storage temperature, with R and Q stable in the solid lipid core. When they are taken into the body, they can be semi-liquid at human body temperature. Hence, the stability of NPs and release of R and Q is controlled by manipulating two Vitamin E derivatives (Zhang L. et al., Clin. Pharmacol. Ther. 2008, 83:761-9; Peer D. et al., Nat. Nanotechnol. 2007, 2:751-60; Nishiyama N., Nat. Nanotechnol. 2007, 2:203-4; Puri A. et al., Crit. Rev. Ther. Drug Carrier Syst. 2009, 26:523-80; Muller R. H. et al., Adv. Drug Deliv. Rev. 2002, 54 Suppl. ES131-55; Teeranachaideekul V. et al., Int. J. Pharm. 2007, 340: 198-206).

[0250] Free bioactive compounds have a low level of bioavailability, because of poor intestinal absorption, reduced distribution, extensive metabolism, and quick excretion. Nanoencapsulation enhances bioavailability via improving absorption, distribution, metabolism and excretion (ADME). Lipid NPs like RQ-NPs can penetrate intestinal epithelial barriers (absorbption) either directly by passive transcellular or paracellular diffusion or indirectly through clathrin-mediated and caveolae-mediated endocytosis followed by transcytosis. A small amount of released R and Q in the enterocytes can also be incorporated into chylomicrons. Both RQ-NPs and chylomicrons are excreted into the mesenteric lymph, and eventually move into the circulation. R and Q delivery to the mesenteric lymphatic system (distribution) has two major advantages: it circumvents the first-pass metabolism in the liver and it to delivers more R and Q to extra-hepatic tissues. In addition, Kolliphor® HS15, a surfactant, is incorporated on the outer layer of NPs. Kolliphor® HS15 contains poly(ethylene glycerol) (PEG) and maintains NPs’ integrity and stability by protecting them from degradation by enzymes and prolong the circulation of NPs by stabilizing them against opsonization. Disease tissues like expanded WAT have leaky blood vessels with enhanced permeability and retention. NPs can easily penetrate and accumulate into these tissues, resulting in delivering more cargos (R and Q) to WAT. Nanoencapsulation protects R and Q from degradation and metabolism. Further, because nanoencapsulated unmetabolized R and Q (hydrophobic) cannot be excreted into the urine, NPs larger than 10 nm cannot be eliminated by kidneys. This results in more RQ-NPs being absorbed into lymph and eventually into the blood stream. Prolonged circulation by PEG coating of NPs enable delivery of more unmetabolized and active R and Q to WAT, resulting in enhanced antiobesity bioactivities (Rein M. J. et al., Br. J. Clin. Pharmacol. 2013, 75:588-602; Su C. et al., Adv. Dru Deliv. Rev. 2019, 143:97-114; Griffin B. T. et al., Adv. Dru Deliv. Rev. 2016, 106:367-380; Neves A. R. et al., J. Colloid. Interface Sci. 2016, 463:258-65; Lundquist P. et al., Adv. Drug Deliv. Rev. 2016, 106:256-276; Rysanek P. et al., Pharm. Res. 2020, 37: 166; Muller R. H. et al., Int. J. of Pharmaceutics, 1996;144: 115-121; Olbrich C. et al., Int. J. Pharm. 1999, 180:31-9; Heurtault B. et al., Pharm. Res. 2002, 19:875-80; Choi H. S. et al., Nat. Biotechnol. 2007, 25: 1165-70).

[0251] It is known that both R and Q improve gut health. Involvement of the gut microbiota and subsequent interindividual differences in the metabolic fate of R may explain some of the persistent gaps in the literature. Polyphenols such as R and Q interact with gut microbiota in reciprocal ways that alter the gut microbial community while also eliciting health benefits to the host. In the presence of a high fat diet, R rescues gut microbiota dysbiosis through decreased abundance of pathogens and enhanced abundance of beneficial bacteria Lactobacillus and Bifidobacterium. Bacteria Slackia equolifaciens and Adlercreutzia equolifaciens convert / ra / i.s-R to dihydroresveratrol, which has been shown to have anti-inflammatory and antiproliferative effects. Resveratrol further improves Bacteroidetes.Firmicutes ratios and reverses the gut microbial dysbiosis induced by a high-fat diet. Supplementing R in a high fat diet in a dose range between 0.2 to 0.4% additionally increases the level of Bacteroides, Lactobacillus, Bifidobacterium, and Akkermausia, and decrease Prevotella, Ruminococcaceae, Anaerotruncus, Alistipes, Helicobacter, Firmicutes, and Peptococcaceae . This causative link between R and gutmicrobiome shift has been recently reviewed with numerous studies pointing to a confirmed role of this shift leading to browning of WAT and reversing of obesity - supporting the coining of the “gut-microbiota-adipose tissue” axis. Recent fecal microbiota transplant studies suggest R- associated microbes enhance insulin sensitivity and decrease weight gain in high mice fed a high fat diet. Similarly, Q can modulate the intestinal microbiota composition and revert gut microbiota imbalance induced by a high fat diet, with subsequent inhibition of inflammasome response in C57BL / 6J mice. Q also improves the Bacteroidetes.Firmicutes ratio, and inhibits growth of Bifidobacterium catemdatum, Lactobacillus spp. and Enterococcus caccae. When Wistar rats were treated with a high fat diet in combination with R (15 mg / kg body weight / day), or Q (30 mg / kg body weight / day), or R + Q, those treated with R+Q had decreased Firmicutes and an improved Bacteroidetes.Firmicutes ratio, lower relative abundance of Desulfovibrionaceae, Acidaminococcaceae, C oriobacteriaceae, Bilophila, Lachnospiraceae, and improved gut microbiota dysbiosis induced by the high fat diet. Aglycones have beneficial effects towards investigated bacteria, while glycosides did not exhibit any effects on intestinal microflora. Encapsulating R and Q into NPs increases their solubility and prevent their metabolism by gut enzymes, which can enhance gut health (Ozdal T. et al., Nutrients. 2016, 8:78; Qiao Y. et al., Food Funct. 2014, 5: 1241-9; Chen M. L. et al., MBio. 2016, 7:e02210-15; Zhou L. et al., Deciphering the Anti-obesity Benefits of Resveratrol: The "Gut Microbiota- Adipose Tissue" Axis. Front Endocrinol (Lausanne). 2019, 10:413; Wang P. et al., Int. J. of Obesity, 44, 213-225, 2020; Wang P. et al., Free Radic. Biol. Med. 2020, 156:83-98; Porras D. et al., Free Radic. Biol. Med. 2017, 102: 188-202; Wicinski M. et al., Nutrients, 2020 Feb, 12(2): 350; Etxeberria U. et al., J. Nutr. Biochem. 2015, 26:651-60).

[0252] Gut microbiota and R and Q have reciprocal effects and the gut microbiota plays a role in transforming R and Q into other metabolites in different gastrointestinal sections, especially in the context of NP delivery systems. This was investigated using both conventional and germ-free C57BL / 6J mice. R and Q metabolism was measured in the liver, kidneys, WAT, BAT and blood. Nutrition and metabolism are closely related, since all metabolites in a biological system come from diet, directly or indirectly. Notably, metabolomics provides a promising approach to study small molecular-weight metabolites and their dynamic changes in biological systems. In particular, mass spectrometry (MS)-based metabolomics has proven to be a highly useful tool for analyzing metabolic alterations that are sensitive to nutrition and diet changes. In an in vivo study using rats, untargeted metabolomics was used for metabolic fecal fingerprinting of R and Q using a high-fat sucrose dietary model. Significantly differing metabolites included carbohydrate derivatives or conjugates in the Q group. The R group was differentiated by compounds related to nucleotides. Moreover, R and R-derived microbial metabolites (dihydroresveratrol and lunularin) were also identified as significant for the classification based on the type of treatment. Interestingly, Zhuang et al. used a rat model to investigate the anti-obesity effects of the combination of Q and R (CQR), in which the ratio of Q and R is 2: 1. They found 22 differential urinary metabolites related to the HFD-induced obesity, which were reversed in a dose-dependent manner by CQR. These key differential metabolites were involved in amino acid metabolism, galactose metabolism, pantothenate and CoA biosynthesis, pyruvate metabolism and lysine degradation. In addition, a study used both transcriptomic and metabolomic profiling approaches show synergistic effects of Q and R supplementation on obesity. The integration of transcriptomic and metabonomic data indicated that Q and R supplementation enhanced processes of glycolysis and fatty acid oxidation, as well as suppressed gluconeogenesis. Based on previous studies, metabolomics is employed to provide holistic and comprehensive insights into the pathophysiological mechanisms of nanoencapsulated Q and R (Zampieri M. et al., Current Opinion in Chemical Biology, 2017, 36: 15-23; Griffin J. L. et al., Nature Reviews Cardiology. 2011, 8:630-643; Nagana Gowda G. A. et al., Curr. Metabolomics. 2013, 1 :227-240; Gu H. et al., Future Oncol. 2012, 8: 1207-1210; Dunn W. B. et al., Chemical Society Reviews. 2011, 40:387-426; Xia J. et al., Metabolomics. 2013, 9:280-299; Fan T. W. M. et al., J. Biomol. NMR. 2011, 49:267-280; Reaves M. L. et al., Curr. Opin. Biotechnol. 2011, 22: 17-25; Patti G. J. et al., Nat. Rev. Mol. Cell Biol. 2012, 13:263- 269; McCartney A. et al., Cancer Treatment Reviews. 2018, 67:88-96; Cui L. et al., Mass Spectrometry Reviews. 2018, 37:772-792; Yan M. et al., Analytica Chimica Acta. 2018, 1037:41-54; Markley J. L. et al., Curr. Opin. Biotechnol. 2017, 43:34-40; Riekeber E. et al., FlOOORes. 2017, 6: 1148-1148; Fiehn O. Curr. Protoc. Mol. Biol. 2016, 114:30.4.1-30.4.32; Etxeberria U. et al., Food Funct. 2015, 6:2758-67; Zhuang T. et al., Front. Pharmacol. 2021, 12:655563; Zhou M. et al., J. Proteome. Res. 2012, 11 :4961-71).

[0253] A nanoencapsulation approach could portend a breakthrough in increasing bioavailability and, promoting gut health and anti-obesity bioactivities of bioactive compounds found in the fruits and vegetables that are regularly consumed. This research helps develop a novel preventive and therapeutic modality for obesity with fewer and less toxic side effects. The outcomes also produce dramatic savings in the cost of medical care and improve the quality of life.

[0254] Biocompatible and biodegradable R-NPs, comprising phosphatidylcholine, alphatocopherol acetate, Kolliphor® HS15 containing PEG, and R, were synthesized. As illustrated in Figure 1 A, phosphatidylcholine is used to form a monolayer of NP membrane. Hydrophilic heads of phosphatidylcholine face outward (towards the aqueous environment), and their two fatty acid tails are buried inside the hydrophobic core of the NPs, and R is encapsulated into the Vitamin E hydrophobic core. The particle size was less than 100 nm in diameter as determined by a transmission electron microscope (TEM), as showin in Figure IB, and a Brookhaven BIMAS particle size analyzer. The encapsulation efficiency of RES was more than 90%, and loading capacity was 22%. R-NPs exhibit a zeta potential of about -26 mV. To examine solubility, 10 mg of R was added into 5 m of 1XPBS, and did not readily dissolve as shown in Figure 1C. R-NPs containing the same amount of R, on the other hand, were completely dissolved in 5 m of 1 xPBS (equivalent to 2 g / L). Therefore, it can be seen that nanoencapsulation dramatically increased R’s aqueous solubility. The particles size of R-NPs did not change at 4 °C and 22 °C for 24 hours, but did increase at 37°C (Figure ID). The poly dispersity index of R-NPs was slightly increased around 15% at 4 °C and 22 °C after 3 days storage (Figure ID), though the zeta potential did not change during storage.

[0255] To determine the anti-obesity activities, mice were fed a high-fat diet for 9 weeks. During weeks 5-9, they were intravenously injected on a biweekly basis with one of the following 4 treatments: saline control (treatment 1), free R (treatment 2), R-NPs (treatment 3), and void (V)-NPs (treatment 4). The dose of R was 52.5 mg / kg body weight at each injection time, hence, the weekly injection dose was 105 mg / kg body weight / week (equivalent 15 mg / kg body weight per day). After sacrificing mice, blood, inguinal WAT (iWAT), and gonadal WAT (gWAT) were collected. As compared to free R, R-NP-treated mice had 10% lower body weight (Figure 2A), 2.5-fold lower fat mass, and 1.5-fold lower % body fat (Figure 2B). Lean mass was similar among all groups (Figure 2C). Since R-NP-treated mice had the lowest body weight, they had the highest % body lean (Figure 2C). Food intake was similar among the four groups (Figure 2D). As compared to free R, R-NP-treated mice had lost 40% gWAT and 25% iWAT (Figure 2E). Hematoxylin and eosin (H&E) staining images (Figure 2F) and quantified adipocyte area data (Figure 2G) indicated that the iWAT adipocyte size from R-NP-treated mice was 1.6-fold smaller than that of free R-treated mice. Further, while R-NPs decreased 50% of fat mass they did not change lean mass after IV delivery.

[0256] R-NPs also deliver more R to WAT and induced iWAT browning. R-NP-treated mice, compared to free R-treated mice, had increased R content in iWAT and gWAT (Figure 3 A), decreased R content in the liver (Figure 3B), more than 2-fold increased iWAT UCP-1 mRNA levels (Figure 3C), and significantly increased iWAT UCP-1 protein levels (Figure 3D). Nanoencapsulation increases the stability of R or other compounds by protecting them from degradation and metabolism. Therefore, intact R-NPs in the circulation are delivered to WAT to exert anti-obesity activity, partially via induced browning of subcutaneous WAT (Zhang J. et al., J. Nutr. Biochem. 2016, 30:14-23; Zhang J. et al., J. Agric. Food Chem. 2013, 61 :9200-9; Zu Y. et al., Colloids. Surf. B. Biointerfaces. 2017, 164:414-423).

[0257] Compared to free R-treated C57BL / 6J mice, those treated with R-NPs had lower blood levels of insulin (Figure 4A) and glucose (Figure 4B), and lower homeostatic model assessment for insulin resistance (HOMA-IR). In parallel to the lowest WAT weight and smallest adipocytes size, R-NPs decreased plasma concentrations of TNF-a, IL-6 and MCP-1 (Figure 4C through Figure 4E) and F4 / 80 expression in iWAT, demonstrating that R-NPs improve glucose homeostasis and attenuate inflammation (Zu Y. et al., J. Control Release. 2021;333:339-351).

[0258] Blood chemistry panels including liver panel (total bilirubin, aspartate amino transferase, alanine amino transferase, alkaline phosphatase, glutamate dehydrogenase), renal panel (total serum protein, albumin, creatinine, blood urea nitrogen), and electrolytes, as well as histological examination for liver, heart, lung, kidneys, brain, spleen, and skeletal muscle, were performed. All reports and data revealed no additional abnormality compared to saline-treatment mice, indicating no toxicity of IV administration of R-NPs for 5 weeks (Zu Y. et al., J. Control Release. 2021;333:339-351).

[0259] Q-NPs were then prepared using a phase inversion-based process. The Q-NPs consist of Q, vitamin E acetate, soy phosphatidylcholine, PEG, surfactant and water. Q-NPs were homogeneous and spherical in shape and about 30 nm in diameter. Their mean particle diameter was around 30 nm in both TEM (Figure 5A) and Brookhaven BI-MAS particle size analyzer measurement. Their poly dispersity index was 0.059. Nanoencapsulation dramatically increased Q solubility in 1 xPBS. When 20 mg of Q was encapsulated into NPs, the Q-NPs were completely dissolved in 10 m of 1 *PBS (equivalent to 2 g / L) at room temperature, showing a translucent state with visible opalescence (Figure 5B). 20 mg of free Q, on the contrary, could not dissolve into the same amount of 1 *PBS at the same temperature (Figure 5B) (Sun M. et al., Colloids SurfB Biointerfaces. 2014;113: 15-24).

[0260] The stability of free Q and Q-NPs in simulated gastric solution (pH=1.2) and simulated intestinal solutions (pH=6.8 and pH=7.4) was measured. Q-NPs had significantly higher stability than free Q in all solutions. Male pre-cannulated Sprague-Dawley rats were given oral gavage of free Q or Q-NPs at dose 50 mg / kg body weight, and blood collected via the catheters at hour 0, 1, 2, 3, 4, 6, 8, 10, 14, 24, 36, and 48 into heparinized tubes. Blood Q concentrations were measured using an HPLC system (Sun M. et al., Colloids Surf B Biointerfaces. 2014; 113: 15-24). Pharmacokinetic parameters were determined on each individual set of data by using a non-compartmental model (Song I. et al., Eur. J. Clin. Pharmacol. 2016;72:665-70). Q-NP -treated compared to free Q-treated rats had 2-fold higher maximum plasma concentration (Cmax), 1.8-fold higher the area under the curve for the concentration-time curve from time zero to time 24h (AUC o^24h), and 2-fold higher area under the curve for the plasma concentration-time curve from time zero to infinity (AUC o— ®) as shown in Table 1. The relative bioavailability Frat infinity was calculated as Fr= AUC Q-NLC, O^ / / AUC Q. < ®. The halflife (ti / 2) = 0.693 / k. The mean residence time (MRT) = 1 / k. Q-NPs had about a significantly 2- fold increase in blood Q concentrations and its relative bioavailability as compared to free Q (Table 1), demonstrating nanoencapsulation improves Q’s pharmacokinetics and bioavailability.

[0261] Table 1. Pharmacokinetic parameters and bioavailability of quercetin after oral administration of free and NP-encapsulated quercetin

[0262] Preparation and characterization of R-NPs, Q-NPs and RQ-NPs: R-NPs, Q-NPs and RQ-NPs are prepared from a lipid mixture composed of 3 mg R and / or 3 mg Q, 6 mg soy phosphatidylcholine, 12 mg a-tocopherol acetate and (±)-a-tocopherol nicotinate, and 12 mg Kolliphor® HS15. After dissolving them in ethanol, ethanol was removed using a nitrogen evaporator. After adding 76°C deionized water, the suspension was homogenized for 1 min followed by sonication for additional 1 min to make R-NPs, Q-NPs and RQ-NPs. V-NPs are prepared using the same method without R and Q. All steps are performed under nitrogen to prevent R and Q degradation. After preparation, all NPs are immediately put on ice. The melting temperatures of a-tocopherol acetate and a-tocopherol nicotinate are 25°C and 38°C, respectively. The hydrophobic core of NPs is solid and stable when the NPs are stored at or below 25°C. The R-NPs, Q-NPs and RQ-NPs are isolated by ultracentrifugation or ultrafiltration (Millipore Amicon Ultra-15). R-NPs, Q-NPs and RQ-NPs are resuspended in lx PBS. The size and morphology of R-NPs are measured using a 200Kv Hitachi H-8100 analytical TEM. The size, size distribution, and zeta potential are measured by using Malvern Zetasizer. R and Q will are detected using a HPLC system (Waters Breeze HPLC system) with a C 18 reverse-phase column and a UV detector. The R mobile phase consists of acetonitrile and 0.2% formic acid in water (30:70, v:v), the flow rate is 1 mL / min. The detection wavelength is 370 nm. The Q mobile phase is acetonitrile / methanol / water (20:60:20, v:v:v) with a flow rate of 1.1 mL / min (Zu Y. et al., J. Control Release. 2021, 333:339-351; Zu Y. et al., Colloids. Surf. B. Biointerfaces. 2018,

[0263] 164:414-423). The detection wavelength is 262 nm. R and Q encapsulation efficiency and loading capacity are calculated according to the following:

[0264] Weight of R / Q added — Weight of free R / Q

[0265] Encapsulation Efficiency (%) = -n— , , , - x 100%

[0266] K 7 7'v 7Weight of R / Q added

[0267] Solubility and stability of R-NPs, Q-NPs and RQ-NPs: The solubility of R and Q in free and nanoencapsulated form dissolved in PBS (pH 7.4) are measured using an HPLC system. The stability of free R and Q, R-NPs, Q-NPs, and RQ-NPs are measured in PBS (pH 7.4) at 4 °C, 22 °C, and 37 °C. The stability is measured in simulated gastric fluid (pH 1.2) and simulated intestinal fluid (pH 6.8 and 7.4) (Ricca Chemical, Arlington, TX) at 37 °C.

[0268] The in vitro release behavior of R and Q is assessed by a dialysis method as previously described. Briefly, free R and Q, R-NPs, Q-NPs, and RQ-NPs are put into two dialysis bags, which are dipped into the dissolution medium composed of 1 *PBS and methanol (9: 1, v:v). The dissolution medium is stirred at 200 rpm at 37 °C, and completely replaced by fresh pre-warmed medium every 2 h. R and Q concentrations in the dissolution medium are measured using the above HPLC methods.

[0269] Pharmacokinetics and biodistribution: Far-red fluorescent lipophilic dye DiD is encapsulated into all NPs. After fasting for 6 h, 144 C57BL / 6J mice are given one of the following 5 treatments: free R (treatment 1), free Q (treatment 2), free F+Q (treatment 3), R-NPs (treatment 4), Q-NPs (treatment 5) and RQ-NPs (treatment 6) dissolved in saline containing 0.1% of Tween 80 via oral gavage. The dose is 80 mg / kg body weight for R and / or Q either as free compound or quantity loaded into the NPs. Twenty-four mice in each treatment group are divided into 8 sub-groups at random for eight sampling points (1, 2, 4, 8, 12, 24, 48, and 72 h after oral administration). Three mice in one sub-group at each time point are humanely sacrificed after their blood is drawn from the abdominal vein. Mice in the time point 24 h group are put into metabolic cages and urine (0-4 h, 5-12 h, and 13-24 h) and feces are collected from these mice. Plasma is obtained by blood centrifugation at l,500xg at 4 °C for 25 min, then aliquoted, frozen and stored at -80 °C until analysis. R and Q concentrations in plasma are determined using an Agilent 1290 UHPLC system. Compartment analysis is used to analyze the data using the standard practical pharmacokinetic program version 97. The following pharmacokinetic parameters of blood, heart, liver, lung, kidneys, spleen, brain, skeletal muscle and GI tract are obtained: area under the concentration-time curve (AUC), half-life (ti 2), total plasma or tissue clearance (Cis), apparent distribution volume (Vd), and rate constant of elimination phase (0).

[0270] The biodistribution of DiD-labeled NPs in mice is measured using an IVIS® Lumina XR imaging system after blood collection. The duodenum jejunum, ileum, cecum, proximal colon, and distal colon are excised and imaged under the IVIS system in the presence or absence of GI lumen content, then they are embedded into an OCT compound. The liver, kidneys, iWAT, gWAT, retroperitoneal WAT (rWAT), interscapular BAT (iBAT) are excised and washed and imaged under the IVIS system, then weighed and stored in OCT and at -80°C until analysis.

[0271] Relevant metabolites, examined via LC-MS / MS, related to Q and R include R-3- O-Sulfate, R-3-O-P-D-glucuronide, dihydroresveratrol, R-4'-O-D-glucuronide, phenylacetic acid, 2-hydroxyphenylacetic acid, 2,4,6-trihydroxybenzoic acid, phloroglucinol, p-hydroxybenzoic acid, etc. These metabolites in the blood, urine and collected tissues are ported to an LC-MS / MS platform. LC-MS / MS experiments are performed on an Agilent 1290 UPLC-6490 QQQ-MS system (Almeida A. F. et al., Compr. Rev. Food Sci. Food Saf. 2018, 17:714-731; Ghu H. et al., Anal. Chem. 2015, 87: 12355-62; Zhu J. et al., J. Proteome. Res. 2014, 13:4120-30; Carrol P. A. et al., Cancer Cell. 2015, 27:271-85; Reyes N. L. et al., Proc. Natl. Acad. Sci. U S A. 2015, 112:424-9; Sperber H. et al., Nat. Cell. Biol. 2015, 17: 1523-35; Ghu H. et al., Angew. Chem. Int. Ed Engl. 2016, 55:15646-15650; Dai D. F. et al., Aging Cell. 2014, 13:529-39; Zhu J. et al., Anal. Bioanal. Chem. 2015, 407:7857-63; Li C. Y. et al., Drug Metab. Dispos. 2018, 46: 1226- 1240; Shao D. et al., Nat. Commun. 2018, 9:2935).

[0272] The cross-sections of duodenum jejunum, ileum, cecum, proximal colon, and distal colon are imaged using a fluorescence microscope with a Cy7 filter. DiD NP signals are quantified using a NIH image J software, providing NP distribution in the GI tract.

[0273] The morphology of different intestinal fractions are examined using a TEM. The cross-sections are stained with hematoxylin and eosin, and evaluated under a microscope. A portion of different intestinal fractions are mounted into modified Ussing Chambers (Physiologic Instruments, Inc.). The permeability is determined by measuring the transmural resistance.

[0274] Measurement of intestinal transport barrier proteins includes mRNA and protein levels of monocarboxylic transporter 1 (Mctl), Mct3, Met 4, Met 5, ZO-1, ZO-2, occludin, claudin, and mucin family and are determined using real-time PCR and immunohistochemistry, respectively.

[0275] Determining R and Q and their metabolites in mesenteric lymph: Thirty male C57BL / 6J mice undergo a surgery for cannulation of mesenteric lymph duct using a PD’s published method with modification using a needled catheter. During the surgery, a feeding tube is put to each mouse. Then the mouse is placed into a Bollman restraint cage, given 5% glucose in saline infusion (0.5 mL / h) via the feeding tube during overnight recovery. After recovery, mice are given the same treatments (n=5) as above followed by saline infusion via the feeding tube. About 200 pL of lymph is collected hourly for 6 h. Blood, liver, kidneys, iWAT, gWAT, rWAT, iBAT are excised and collected, and NP signals in lymph and tissues are determined using the IVIS system (Wang S. et al., J. Nutr. 2006, 136:2791-6; Wang S. et al., J. Nutr. Biochem. 2006, 17:492-8; Banan B. et al., J. Surg. Res. 2021, 260:399-408).

[0276] The size of chylomicrons in lymph are between 200 - 1,200 nm, but the NPs are less than 100 nm. Chylomicrons and NPs are collected using a Superose GL column. DiD dye in the non-particle fraction, and the fractions of chylomicrons and NPs, are measured using an IVIS system. R, Q, and their metabolites in lymph, non-particle fraction, fractions of chylomicrons, and NPs are measured using targeted LC-MS / MS.

[0277] R-NPs, Q-NPs and RQ-NPs improve solubility and stability of R and Q. More NPs are distributed into blood, lymph, and adipose tissues, and dramatically increase their R and Q concentrations, especially unmetabolized R and Q. NPs are transported in lymph, then into the blood stream. NPs do not have adverse effects on gut integrity and permeability.

[0278] Determining anti-obesity bioactivities of RQ-NPs in C57BL / 6J mice: The effects of R and Q via NPs delivery are apparent in C57BL / 6J mice (a commonly used high-fat diet induced obesity animal model), in terms of body weight and fat mass reduction. The high fat diet induces obesity, which is reduced by the intervention with RQ-NPs. The anti-obesity mechanisms are determined by measuring lipid content; adipocyte size and mitochondria amount; fatty acid P-oxidation and mitochondrial respiration; and mRNA and protein expression of genes involved in adipogenesis, lipogenesis, lipolysis in the liver and adipose tissue, which are correlated with more unmetabolized R and Q in the blood, adipose tissue depots. Since NPs are composed of biocompatible and biodegradable components, there are no associated toxicides to organs or tissues. Routine necropsy, blood test and histological examination for the liver, heart, lung, kidneys, brain, spleen, GI tract and skeletal muscle are performed.

[0279] After one week of acclimation, 128 male C57BL / 6J mice (Jackson Laboratory) are weighed and randomly assigned to one of eight groups (16 mice / group). Mice receive one of the eight treatments (Figure 6) via mixing into the semi-purified high fat diet.

[0280] Primary outcomes are body weight and percent body fat; there are 12 pairwise considerations of 8 treatment groups over these two outcomes. In a one-way ANOVA study, a sample of 128 mice, divided among 8 groups, achieves a power of no less than 80% to detect a given between-group difference if the true effect size is at least 1.5. This power assumes an F test is used with a significance level of 0.05. C57BL / 6J mice are fed a high fat diet (fat providing 45% of energy) prepared by following the D12451 formula from Research Diets. Since all NPs contain vitamin E, less Vitamin E is added in the four NP treatment groups to ensure that total vitamin E content in the diet is the same among all groups. Since the NPs are solid at room temperature, NPs are mixed with diet components. R or Q will supplement 0.1% of diet (RQ treatments contain 0.1% of each), that is equivalent about 80 mg / kg body weight administration for R or Q. The dose is in the commonly used dose range of R and Q. Mice are allowed to drink and eat the high fat diet ad libitum. Mice receive treatments for 8 weeks with food intake and body weight measured biweekly. At week 8, mice are placed into metabolic cages for collecting 24-h urine and feces. After 8 weeks, mice are fasted (6 h), humanely sacrificed, and complete necropsies are performed. Blood is collected from the abdominal artery under anesthesia and lymph is collected from the mesenteric lymph duct. Liver, lung, spleen, skeletal muscle, iWAT (subcutaneous), gWAT and rpWAT (visceral), iBAT, kidneys, brain, heart and GI tract of each mouse are collected, measured, weighed and described in detail. Each tissue is cut into 3 pieces to be immediately frozen in liquid nitrogen followed by storage at -80 °C; embedded in OCT compound; and fixed in 4% paraformaldehyde (for histology).

[0281] Body composition is measured using Echo-MRI weekly. Under anesthesia, transponders are implanted into the abdominal cavity of mice before the study. Core body temperature is measured for 24 h each week. Meanwhile, mice are put into CLAMs cages for 24 h on the same days. Respiratory quotient and total energy expenditure are calculated as in Weir (Weir J. B. et al., J. Physiol. 1949, 109: 1-9).

[0282] On the day of sacrifice (week 8), at least 1 mL of blood is collected, half of which is used for safety evaluation. The remaining blood is centrifuged at 2,000 x g at 4 °C for 20 min. Plasma concentrations of free fatty acids, triglycerides, TC, LDL-C and high-density lipoprotein (HDL)-cholesterol are measured by an enzymatic method as described previously (Wang S. et al., Atherosclerosis. 2009, 204: 147-55). Non-HDL-chol esterol = TC - HDL-cholesterol.

[0283] Mice are fasted for 6 h at weeks 0, 4 and 8. Fasting blood is collected from tail veins at week 0 and 4 and from the abdominal vein at week 8. Plasma is collected by blood centrifugation. Blood glucose and insulin (Millipore) concentrations are measured using commercial kits. The homeostasis model assessment of insulin resistance (HOMA-IR) is calculated using fasting blood glucose and insulin concentrations. The following calculation is used to estimate insulin resistance: HOMA-IR = fasting glucose (mmol / L) x fasting insulin (mU / L) / 22.5.

[0284] Glucose tolerance tests (GTT) and insulin tolerance tests (ITT) are performed at the end of treatments. For GTT, mice are fasted for 6 h and then injected intraperitoneally with glucose at a dose of 1 mg / kg body weight. Blood is collected from the tail snip and blood glucose measured with a glucometer at 0, 15, 30, 60, 90, and 120 min post injection. To measure ITT, mice are fasted for 6 h and then injected intraperitoneally with insulin at a dose of 0.75 U / kg body weight. Blood glucose is measured as well.

[0285] Plasma is obtained for measurement of inflammatory and anti-inflammatory factors, adiponectin, and leptin. The liver, iWAT, and gWAT are homogenized, and after centrifuging, the supernatant is collected. Concentrations of IL-la, IL-ip, IL-6, IL-10, IL-12p70, IL-17A, IL-23, IL-27, MCP-1, IFN-P, IFN-y, TNF-a, and GM-CSF in the plasma, liver, iWAT and gWAT are measured using a LEGENDplex ™ mouse inflammation panel kit and flow cytometry. Adiponectin and leptin are determined using the MesoScale Multiplex and ELISA kits (R&D Systems).

[0286] Macrophage infdtration is analyzed by immunohistochemistry analysis of macrophage marker (e.g., mac-2) on the liver and WAT tissue section slides. Liver and WAT lipid are extracted and liver and WAT triglyceride and cholesterol content are measured as described in the literature (Wang S. et al., Clin. Nutr. 2014, 33:260-6; Wang S. et al., Nutr. Res. 2013, 33: 1072-82)

[0287] Adipocyte size, macrophage infiltration and mitochondria are also measured. Sections of WAT and BAT are stained using hematoxylin-eosin. Mean diameter is calculated using NIH Image J software. Staining of Ml- and M2- type macrophages is performed using an immunohistochemistry method. Frozen sections of WAT and BAT are fixed and stained using MitoTracker Green FM dye (Life Technologies). The amount and location of mitochondria in white and brown adipocytes is visualized under a fluorescent microscope (EVOS FL Auto).

[0288] Fatty acid oxidation and mitochondrial respiration are measured using Seahorse. Mitochondrial DNA are measured using real-time PCR.

[0289] RNA and proteins are extracted from the liver and WAT using standard methods. cDNA synthesis and real-time PCR are conducted as described previously. Protein levels are measured by Western blotting or immunohistochemistry as described previously. Genes of interest in WAT include GLUT4, ADRB3, UCP1, UCP2, UCP3, PPARy, PGC-la, PRDM-16, SIRT1, SCD1, FABP4, FASN, HMGCR, ACC1, GAPDH, CPT, Cidea, PRDM16, Elovl3, CPTla, MCAD, Tmeme26, DIO2, and HSL, with 36B4 used as an endogenous control. Genes of interest in the liver include TNFa, MCP1, TLR4, NLRP3, HSF, SOCS4, and IL-6, with P- actin used as an endogenous control.

[0290] Untargeted aqueous metabolomics are used to examine broad changes in hydrophilic metabolome. The high sensitivity of LC / GC-MS methods detect >1,200 aqueous metabolites / features in a typical biological sample. Untargeted profiling is performed using Agilent 7820 GC-5977 MS and Thermo Scientific Electron Orbitrap Velos Pro UPLC-MS instruments in the blood, liver iWAT, gWAT and iBAT. To identify peaks from the MS spectra, in-house chemical standards (-400 aqueous metabolites), commercial Agilent database (Mass Profder Professional Software, already in Gu’s lab), robust NIST and Fiehn’s libraries, the HMDB metabolite library, and the METLIN database are used (Qi Y. et al., Evid. Based Complement. Alternat. Med. 2013, 2013:438680; Yao W. et al., Anal. Bioanal. Chem. 2014, 406:7367-78; Buas M. F. et al., Gynecol. Oncol. 2016, 140: 138-44; Babushok V. I. et al., J. Chromatogr. A. 2007, 1157:414-21; Wishart D. S. et al., Nucleic Acids Res. 2013, 4ED801-7; Tautenhahn R. et al., Nat. Biotechnol. 2012, 30:826-8; Wishart D. S. et al., Nucleic Acids Res. 2018, 46:D608-d617).

[0291] Untargeted lipidomics profiling is used to examine broad changes in lipids. Lipidomics focuses on the lipid metabolism, and it has become an important approach to describe the metabolic differences induced by different biological status. Global lipid changes in in the blood, liver iWAT, gWAT and iBAT are investigated using the Thermo Scientific Electron Orbitrap Velos Pro UPLC-MS system. In a typical run, more than 400 identified lipid compounds are observed, comparing the obtained >1,500 MS features against the in-house chemical standards (-550 lipids) and, public databases such as the Lipidmaps database (Briigger B., Annual Review of Biochemistry. 2014, 83:79-98; Hsu F-F., Analytical and bioanalytical chemistry. 2018, 410:6387-6409; Newsholme P., Essays in Biochemistry. 2016, 60:407-407; Chen X. et al., Oncotarget. 2016, 7:36622-36631; Sethi S. et al., Prostaglandins & Other Lipid Mediators. 2017, 128-129:8-16; Wenk M. R., Nature Reviews Drug Discovery. 2005, 4:594- 610).

[0292] Targeted LC-MS / MS is used to examine R and Q metabolites of interest. R and Q, and their metabolites in blood, lymph, urine, feces, and tissues (liver, iWAT, gWAT, iBAT, kidneys, spleen, duodenum jejunum, ileum, cecum, proximal colon, and distal colon) of each mouse are determined using the targeted LC-MS / MS as described.

[0293] Intestinal histomorphology and permeability and intestinal transport barrier proteins are measured. These assays are conducted as previously described to compare the effect of short-term and long-term treatments on gut integrity.

[0294] After 8 weeks of treatment, half of the mice from each treatment group are randomly selected for safety evaluation. Complete necropsies are performed on the day of sacrifice, including full gross and microscopic evaluation of all organ-systems. After terminal exsanguinations under isoflurane, heart, liver, lung, kidneys, skeletal muscle, brain, GI tract, and spleen of each mouse are collected, measured, weighed and described in detail. They are fixed, embedded, sectioned, and stained for histological examination and evaluation. Blood is used for hematology and clinical chemistry.

[0295] RQ-NPs compared to other treatment groups significantly reduce body weight and fat mass of C57BL / 6J mice via increasing lipolysis and fat oxidation in WAT, decreasing adipogenesis and lipogenesis in WAT and liver, activating thermogenesis, browning iWAT, reducing inflammation, that are correlated with more unmetabolized R and Q in WAT depots. There are no associated toxicities to organs or tissues. Both R and Q in RQ-NPs have synergistic effects on exerting the above beneficial effects.

[0296] Nanoencapsulation dramatically increases R / Q solubility and prevents them from metabolism by enzymes. Even though more RQ-NPs are absorbed in the small intestine, RQ-NPs compared to free R / Q still deliver more soluble and unmetabolized R / Q in the lower small intestine, cecum and large intestine, which has beneficial effects on the gut microbial community structure and relative abundance of specific microbial taxa. The gut microbiota and host metabolize free and nanoencapsulated R / Q differently, and these differences in metabolites throughout the gut coincide with gut microbiota composition, and anti-obesity effects.

[0297] The gut-microbiota of conventionally-raised mice fed RQ-NPs is profiled. At 8 weeks of age, small and large intestinal contents are collected from 80 of the C57BL / 6J mice (n=10 per group). Samples are isolated into separate fractions based on intestinal location (duodenum, jejunum, ileum, cecum, proximal colon, and distal colon). Contents from each location are snap frozen at -80 °C in sterile tubes until further processing. Microbial DNA is extracted and isolated from the intestinal contents using DNeasy® PowerSoil® Pro DNA Isolation Kits (Qiagen, Hilden, Germany) according to manufacturer protocol using a vortexbased beadbeater. Bacterial community analysis is performed via next generation sequencing using the MiSeq Illumina platform (San Diego, CA). Amplicon sequencing of the V4 region of the 16S rRNA gene is performed using the barcoded primer set 515f / 806r and 5 Prime Hot Master Mix (5 Prime, Germany), as described by the Earth Microbiome Project (EMP). PCR amplifications for each sample are performed in triplicate, and DNA amplicons pooled and quantified using the Quant-iT™ PicoGreen® dsDNA Assay Kit (ThermoFisher, Waltham, MA). A no-template control sample is included during library preparation as a control for extraneous nucleic acid contamination. A total of 200 ng of DNA from each sample is pooled and then cleaned using the QIAquick® PCR purification kit (QIAGEN, Hilden, Germany). The pooled sample is quantified using the Illumina library Quantification Kit ABI Prism® (Kapa Biosystems, Wilmington, MA) and diluted to a final concentration of 4 nM. The sample is further denatured and diluted to a final concentration of 4 pM with 15% PhiX. Finally, the DNA library is loaded in the MiSeq Illumina (SN M02149, with the MiSeq Control Software v 2.5.0.5) and run using the version 2 module, with 2 x 250 paired-end reads, and following the directions of the manufacturer.

[0298] Pooled sequences are de-multiplexed and quality filtered using the QIIME2 software package (Bolyen E. et al., Nat. Biotechnol. 2019;37:852-857). Sequences are assigned to amplicon sequence variants (ASVs) with a 99% similarity threshold using QIIME’s uclust- based open-reference ASV picking protocol against most recent SILVA or GreenGenes reference databases. Sequences that do not match the reference database are clustered de novo; thus, all sequences are included in the analysis. Core diversity analyses are performed on the ASV tables, including alpha and beta diversity as well as taxonomic summaries as provided by QIIME2 software.

[0299] Forty 8-week-old C57BL / 6 germ-free mice are fed the same treatment as described (n=5) to discern microbial vs. host metabolisms related to the compounds delivered. Diet is formulated to be identical as the high fat diet in macronutrient composition prior to double gamma irradiation to maintain sterility. However, 1.5x vitamin mix is used to account for micronutrient loss during irradiation. The germ-free mice from Co-I Cross’ breeding colony is housed in separate gnotobiotic cages through a Sentry Sealed Positive Pressure system. Mice receive one of the eight treatments via drink bottles daily. After 2 weeks, intestines (duodenum, jejunum, ileum, cecum, proximal colon, and distal colon), blood, liver, kidneys, spleen, adipose tissue depots (iWAT, gWAT, iBAT) are collected and the profiles of R and Q metabolites are assessed in each intestinal segment and tissue and blood. Further, at time of euthanasia, a sterility test is performed in cecal content to confirm axenic status using multiple methods including anaerobic and aerobic culturing, gram-staining, and PCR.

[0300] Data is checked for normality, and appropriate transformations are performed when necessary prior to statistical analysis (SPSS). The results are analyzed by Student’s t-test for two groups, and an analysis of variance followed by Tukey’s post hoc test is performed to compare multiple group means. Differences are considered significant at / ? < 0.05. Each data point is measured in triplicate, and each experiment is performed at least three times independently wherever possible. Microbiome composition, alpha- (observed ASVs, phylogenetic diversity, and taxa evenness) and beta-diversity (UniFrac phylogenetic distance between samples) metrics are calculated in Qiime 2 (Quantitative Insights into Microbial Ecology 2). Random forest-based regression models are used to evaluate the relative rates of microbial change over time in response to treatment; microbiota composition is considered the predictor of animal age in weeks with treatment group as an independent variable. Significant differences in beta-diversity are evaluated using permutational MANOVA models, while longitudinal changes in alpha-diversity and microbiota change over time are examined by piecewise linear model tree analysis. Significant differences in the relative abundance at each time point are evaluated using LEfSe to identify microbial biomarkers that correlate with treatment-induced anti-obesity outcomes. To correct for multiple comparisons, P values of the estimated treatment effect are adjusted to false discovery rates (FDRs) (Lozupone C. et al., Appl. Environ. Microbiol. 2005, 71 :8228-35; Bolyen E. et al., Nat. Biotechnol. 2019, 37: 1091;

[0301] Breiman L., Machine Learning. 2001, 45:5-32; G. M. Fitzmaurice NML, J. H. Ware, Applied Longitudinal Analysis, Hoboken, NJ; 2004; Segata N. et al., Genome Biol. 2011, 12:R60).

[0302] This innovation is a potential breakthrough in combating obesity by using bioactive R and Q with maximized efficacy and minimized immunogenicity and side effects. It provides new insights and advance knowledge on bioactive compounds, nutrition, and obesity, as well as dramatic savings in the cost of medical care.

[0303] Examining anti-obesity effects ofRQ-NPs in human fecal microbiota transplant C57BL / 6J mice In addition to the anti-obesity effects of nanosized resveratrol and quercetin (RQ- NPs), the underlying mechanisms of phytochemicals (such as resveratrol and quercetin) is of interest. Trans-resveratrol (R) and quercetin (Q), have displayed numerous health advantages, including anti-obesity effects. However, their low solubility and stability limit their application. Encapsulating trans-resveratrol (R) and quercetin (Q) into nanoparticles (RQ-NPs) can overcome these issues. Gut microbiota (GM) also play a pivotal role in obesity development, and transresveratrol and quercetin metabolism (Figure 7).

[0304] Recent dietary intervention and obesity-related research largely concentrates on the thermogenesis activation of adipose tissues and gut microbiota regulation. Adipose tissues are essential for maintaining temperature homeostasis because they boost heat generation, which is linked to higher fatty acid and glucose burning and therefore reduce body weight. Growing evidence also suggests that the gut microbiota is linked to glucose metabolism and energy balance, both of which are also linked to obesity development. In addition, “humanizing” the gut of antibiotic-treated mice via healthy human fecal transplantation (HFT) provides insights into understanding of how the GM can regulate human health. (B. B. Lowell et al., Nature 404(6778) (2000) 652-60; M. Cedikova et al., Stem Cells Int. (2016) 6067349; P. D. Cani et al., Nat. Rev. Endocrinol. 10(2) (2014) 74-6).

[0305] The characteristics of RQ-NPs are investigated and evaluated for their antiobesity effects on C57BL / 6J mice with or without HFT, establishing their connection with gut microbiota alteration, and determine their relationships with thermogenesis activation.

[0306] To investigate the anti-obesity effects of resveratrol + quercetin nanoparticles on high-fat-diet-fed C57BL / 6J obese mice, and weekly body weight changes are monitored. In addition, specific physiological changes that accompany weight loss are also investigated, such as insulin / glucose tolerance test, energy expenditure and plasma profiles. It is believed that RQ- NPs serve as an effective therapeutic intervention for promoting weight loss.

[0307] The role of adipose tissue thermogenesis activation and mice original GM and HFT-treated mice GM in the beneficial effect of RQ-NPs intake is investigated. This is achieved by evaluating changes in mice gut microbiota homeostasis and investigating thermogenesis- related signaling pathways regulations in RQ-NPs treatment groups. RQ-NPs could combat obesity by activating thermogenesis and regulating gut microbiota compositions (favoring “good” strains and inhibiting “bad” strains). The outcomes yield references for future application of nanoencapsulated resveratrol and quercetin for the treatment of obesity.

[0308] The amplified mucoadhesion resulting from the application of NPs bears the advantage of refining the oral delivery of inadequately adsorbed drugs, proteins, and nucleotides, by prolonging the duration and magnitude of their interaction with the intestinal mucus layer. Previous studies demonstrated that both R and Q can be easily dispersed and encapsulated in the hydrophobic core of the lipid NPs with improved stability and beneficial activities (Y. Zhu et al., Colloids Surf B Biointerfaces 2018 Apr 1 : 164:414-423; M. Sun et al., Colloids Surf B Biointerfaces, 2014 Ian 1 :113: 15-24; L. M. Ensign et al., Adv Drug Deliv Rev 64(6) (2012) 557- 70; J. M. Gamboa et al., Adv Drug Deliv Rev 65(6) (2013) 800-10; Y. Zhu et al., J Control Release 2021 May 10:333:339-351).

[0309] According to previous research, dietary intervention with the abovementioned R and Q activates brown adipose tissues (BAT) and induces the browning of white adipose tissues (WAT). Previous research also demonstrated that WAT browning and BAT activation may play a significant role in preventing the development of obesity by boosting body thermogenesis. The uncoupling protein 1 (UCP1 an important marker of browning, enhances adipose tissue thermogenic activity. Additionally, WAT browning is accompanied by an increase in the mRNA expression level of the PPARg-coactivator- la (PGC-1 ). However, many markers are involved in the thermogenesis process and the underlying mechanisms of R and Q remain incompletely understood, especially for nanosized R and Q. Elucidating the complex mechanism that regulates the thermogenesis of BAT and WAT provides future approaches for treating obesity. Thus, further research is being conducted to determine the relationship between dietary intervention with NPs, obesity, and thermogenesis (J. M. Andrade et al., Eur J Nutr 53(7) (2014) 1503-10; Y. Pei et al., J Nutr Biochem 88 (2021) 108532; N. Arias et al., Obesity (Silver Spring) 25(1) (2017) 111-121; P. Cohen et al., Mol Biol Cell 27(16) (2016) 2523-7; L. Vergnes et al., J Biol Chem 295(44) (2020) 15054-15069; A. Roesler et al., Biochem J 477(3) (2020) 709-725; R. H. Freire et al., International Journal of Obesity 40(3) (2016) 479-486; X. Liu et al., Biochem Biophys Res Commun 545 (2021) 189-194; J. Liang et al., Diabetes Metab Syndr Obes 2021 Oct 27: 14:4351- 4360).

[0310] There is need for elucidating the underlying anti-obesity mechanisms related to the gut microbiota. Previous studies suggest that altering the gut microbiota offers an innovative therapeutic target for obesity. Additionally, dietary intervention influences gut microbiota composition, which subsequently affects human metabolism. More research is needed to ascertain the connections between the regulation of the gut microbiota and dietary interventions involving RQ-NPs. Thus, this work is addressing this need and could inform new treatment alternatives for combating obesity disease. It is hypothesized that NPs treatment may alter the gut microbiota composition, and then improve its related metabolic syndromes, which may be correlated with hindered obesity development (P. J. Turnbaugh et al., J Physiol 587(Pt 17) (2009) 4153-8; A. F. Pihl et al., Child Obes 12(4) (2016) 292-9; R. K. Singh et al., Transl Med 15(1) (2017) 73; G. K. John et al., Genes (Basel). 2018 Mar: 9(3): 167; X. Zhang et al., Geroscience 43(2) (2021) 709-725).

[0311] NPs are optimized using vitamin E as a hydrophobic core for protection of resveratrol and quercetin, and their solubility, stability, bioavailability, and anti-obesity activity improvement. Encapsulation of resveratrol and quercetin in NPs comprised of soy phosphatidylcholine (PC), Kolliphor® (polyethylene glycol (15)-hydroxy-12-stearate), and alpha-tocopherol acetate (the primary form of vitamin E) with an appropriate ratio (4:7:22:22), significantly enhances their solubility and stability.

[0312] To leverage prior work and advance NPs technology, the design of RQ-NPs is optimized and their physical stability (size, polydispersity index, and zeta potential) and chemical stability (encapsulation efficiency) are investigated first. Preliminary findings on resveratrol (R) indicate that encapsulating R within NPs composed of soy PC, aTA, and Kolliphor® substantially augment its aqueous solubility and chemical stability. Notably, while 0.75 mg of free R failed to completely dissolve in 1 mL of 1 x PBS at room temperature, R-NPs incorporating the same amount of resveratrol manifested a translucent state with discernible opalescence (Figure 8A). Furthermore, an / / / vitro analysis of the release pattern of R from R- NPs was conducted and compared it to that of free R using a dialysis method. The results revealed that free R exhibited a burst release pattern, with 90% of R being released within the first 2 hours (Figure 8B). On the other hand, R-NPs released only around 40% of R within the first 2 hours, thus demonstrating the potential of nanotechnology to significantly enhance the stability of R. Moreover, in vivo experiments showed that R-NPs had a more potent anti-obesity effect as compared to free R (Figure 8C). Preliminary findings on quercetin (Q) similarly indicate that synthesizing Q-NPs can enhance its water solubility by at least 1000-fold, and showed a sustained release pattern (Figure 8B).

[0313] Those observations led to further investigation into the applications of R-NPs, Q- NPs, and combined RQNPs in the area anti-obesity. The long-term goal of this research is to provide new avenues for tackling obesity by leveraging the potential of nutritional phytochemicals and NPs technology and expanding the scope of natural R and Q applications in the domain of obesity therapy. The primary hypothesis of this proposal is that RQ-NPs may represent a targeted and efficacious therapeutic intervention for treating obesity, through the modulation of gut microbiota and the thermogenesis stimulation of adipose tissue.

[0314] High-fat-diet (HFD)-fed C57BL / 6J obese mice with and without healthy human fecal transplantation (HFT) are used to investigate the role of GM in the beneficial anti-obesity effects of RQ-NPs (Figure 9). Male C57BL / 6J mice (6 weeks old) are acclimatized for a week and fed a low-fat diet (LFD) with ad libitum access to water. Afterward, they are randomly assigned to one of 7 groups: LFD (10% of calories derived from fat), high-fat diet (HFD, 45% of calories derived from fat), HFD + HFT, free RQ + HFD, free RQ + HFD + HFT, RQ-NPs + HFD, RQ-NPs + HFD + HFT. R and Q are orally administered to mice three times per week at 10 mg / kg body weight for 10 weeks. According to prior research, administering free R to obese Zucker rats at a lower dosage of 10 mg / kg per day for a duration of 8 weeks results in a significant decrease in both serum free fatty acid levels and abdominal fat content. Furthermore, another study indicated that administering Free R treatment to high-fat diet (HFD) mice at a dosage of 5 or 10 mg / kg for a period of 17 weeks significantly prevented the development of obesity. Thus, in this study, 10 mg / kg body weight dose is selected for comparing the antiobesity effects of free RQ and RQ-NPs. Sterile cages are changed every day to avoid coprophagy in rodent models. Body weight and composition are measured weekly. At the end of the experiment, mice are fasted for 16 hours, euthanized, and their blood, livers, BAT, and WAT immediately removed, weighed, and preserved. Tissues are sectioned after being embedded in the CRYO-OCT compound, and tissue sections (7 pm) are prepared and stained with hematoxylin and eosin (H&E) for general morphological observations (L. Rivera et al., Biochem Pharmacol 77(6) (2009) 1053-63; J. P. Huang et al., J Cardiovasc Pharmacol 72(5) (2018) 231- 2410).

[0315] Mice are treated with an antibiotic cocktail consisting of 0.5 g / L vancomycin, 1 g / L neomycin, 0.1 g / L metronidazole, and 1 g / L amphotericin-B for 2 weeks in autoclaved drinking water ad libitum. Flavoring (20 g / L grape flavored Kool-Aid Drink) is added in the autoclaved drinking water for all groups. Freshly voided fecal samples are collected weekly.

[0316] Twelve hours after the final antibiotic gavage, mice in the fecal transfer treatments are gavaged with 150 pL fecal material reconstituted in sterile saline from human donors. Mice in the weekly fecal transfer treatments are gavaged weekly for 7 weeks with fecal material from human donors.

[0317] At the end of the treatment, a glucose tolerance test (GTT) is conducted. Baseline glucose values (0 min) are measured by collecting blood samples from the tail vein prior to the injection of glucose (1.5 g / kg body weight). Additional blood samples are collected at regular intervals of 15 min, 30 min, 60 min, 90 min, and 120 min.

[0318] Mice's energy expenditure (EE) is determined using the CLAMS. The system continuously records O2 consumption (FO2, mL / kg / h), CO2 production (ECO2, mL / kg / h), and physical activity over a 24-hour period. The following formula is used to calculate the respiratory exchange ratio (RER) and EE.

[0319] VcoRER = Vo23.815 + 1.232E, EE - -v. - x Vo

[0320] Vco2

[0321] High-density lipoprotein (HDL), low-density lipoprotein (LDL), total cholesterol (TC), and triglycerides (TG) are measured using kits from Sigma and Cayman after collecting the samples. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are measured using kits from Sigma after the samples are collected.

[0322] Obesity arises from a metabolic disorder caused by disturbed energy homeostasis, where energy intake exceeds energy expenditure. In recent years, studies also reported that activating BAT heat production could provide a potential alternative method for regulating the energy balance. Additionally, the formation of beige cells in WAT, also referred to as "WAT browning", is essential in increasing EE. Research has also shown that certain stimuli, including chronic cold exposure, exercise, and consumption of nutritious foods, can modulate BAT activation and WAT browning. Therefore, the administration of RQ-NPs exhibits their antiobesity properties by stimulating brown adipose tissue activation and inducing white adipose tissue browning in HFD-induced obese mice (R. Zhu et al., Food Chemistry 141(1) (2013) 482- 487; J. Villarroya et al., J Endocrinol. 2019 Nov:243(2):R19-R27; K. A. Virtanen et al., N Engl J Med 2009:360: 1518-1525; M. Harms et al., Nature Medicine 19(10) (2013) 1252-1263; J. Nedergaard et al., Cell Metabolism 20(3) (2014) 396-407).

[0323] In addition to the thermogenic activity of adipose tissues, the mammalian gut microbiome also exerts a significant influence on the host's well-being, encompassing energy metabolism and regulation of body weight. Many studies in mice have suggested that dysbiosis, or imbalance, of the gut microbiota, is linked with the initiation, progression, and aggravation of various disorders, such as obesity. Moreover, research also indicated that diet intervention plays an important role in regulating GM. Given this association, GM also plays an important role in regulating obesity development in HFD-fed mice supplemented with RQ-NPs (P. J. Turnbaugh et al., Nature 444(7122) (2006) 1027-31; V. K. Ridaura et al., Science 341(6150) (2013) 1241214; C. Chang et al., Best Pract Res Clin Gastroenterol 30(1) (2016) 3-15; H. Xu et al., J Immunol Res 2019 (2019) 7546047; H. Tilg et al., Gastroenterology 136(5) (2009) 1476-83).

[0324] At the end of the experiment, four intestinal regions (duodenum, jejunum, ileum, and colon) are isolated and tissue sections (7 pm) are prepared and stained with hematoxylin and eosin (H&E) for histological comparison. GM from small intestinal contents, caecal contents, and large intestinal contents are collected for future GM analysis.

[0325] Bacterial community analysis is performed via next-generation sequencing in the MiSeq Illumina platform. Amplicon sequencing of the V4 region of the 16S rRNA gene is performed with the barcoded primer set 515f / 806r designed by Caporaso et al. 2011 and following the protocol by the Earth Microbiome Project (EMP) for the library preparation. PCR amplifications for each sample are done in duplicate, then pooled and quantified using an Accublue kit. A no-template control sample is included during the library preparation as a control for extraneous nucleic acid contamination. 240 ng of DNA per sample are pooled and then cleaned using QIA quick PCR purification kit (QIAGEN). The pool is quantified by using the qubit. Then, the DNA pool is diluted to a final concentration of 4 nM then denatured and diluted to a final concentration of 4 pM with a 25% of PhiX. Finally, the DNA library is loaded in the MiSeq Illumina and run using the version 2 module, 2x250 paired-end, following the directions of the manufacturer.

[0326] The evaluation of beta-diversity is calculated through the R mocroeco package and depicted through principal coordinate analysis (PCoA).

[0327] The mRNA levels and protein levels of intestinal lipid and glucose transport- and intestine integrityrelated genes, and the adipose tissues lipid metabolism- and thermogenesis- related genes (Table 2). The extraction of total RNA from the adipose tissues (BAT and WAT) and intestinal tissues (duodenumjejunum, ileum, and colon) are performed, and the subsequent RT-qPCR analysis and data analysis are conducted following established protocols. Furthermore, the investigation of Western blot determinations of specific pathways are conducted (F. Zhou et al., lournal of Functional Foods 62(July) (2019) 103509-103509).

[0328] Statistical analysis is conducted using SPSS (version 28.0, IBM Corp., Armonk, NY). One-way analysis of variance (ANOVA) is used to evaluate the significance of differences between groups and Bonferroni statistical test are applied as a Post Hoc, and p < 0.05 is considered statistically significant.

[0329] 'able 2. Genes investigated in this study

[0330] RQ-NPs were found to decrease body weight, fat mass and liver inflammation, and improve glucose intolerance of obese mice as shown in Figure 10. RQ-NPs increase thermogenesis and gut integrity-related gene expression as shown in Figure 11. RQ-NPs alter the microbiota composition at the phylum level, which increases the relative abundance of Bacteroidetes, and decreases the relative abundance of Verrucomicrobia in HFT-treated mice as shown in Figure 12. In addition, RQ-NPs changed the beta-diversity of high-fat diet-treated mice’s gut microbiota (Figure 13 A). Human fecal transplantation itself engrafted the mice with a different gut microbiota composition (Figure 13B), and RQ-NP treatment further altered the gut microbiota composition of HFT-treated mice (Figure 13C).

[0331] Compared to Free RQ, RQ-NPs treated mice had: decreased body weight and fat mass, increased EE and expression of thermogenic genes, lowered liver inflammation-related gene expression, and activated thermogenesis- and gut integrity-related gene expressions. Both RQ-NPs and Free RQ increased relative abundance of Bacteroidetes in HFT-mice, while decreasing the relative abundance of Verrucomicrobia in HFT-mice. RQ-NPs decreased body weight and fat mass, improved glucose homeostasis, improved liver function, enhanced gut integrity, and optimized microbiota composition of obese mice.

[0332] Compared to the control, mice treated with RQ-NPs, with or without HFT, had 17% and 21% lower body weight, respectively, and had 38% and 30% lower fat mass, respectively. Increased EE was reflected by increased O2 consumption and CO2 release in RQ- NP -treated mice with or without HFT. Compared to the control mice, RQ-NP -treatment with or without HFT showed 32% and 25% lower area under the curve during GTT. RQ-NP -treated mice with or without HFT compared to the control mice also had 90% and 80% increase in HDL levels, and 13% and 15% decrease in LDL / VLDL levels, respectively. RQ-NP treatment with or without HFT decreased blood AST and ALT levels that were correlated with lowered expression of liver inflammatory factor genes. Results also indicated that RQ-NPs with or without HFT- treated groups showed better anti-obesity effects compared with free RQ with or without HFT- treated groups in these measurements. RQ-NPs enhanced expression of gut integrity genes, and increased relative abundance of Bacteroidetes of obese mice.

[0333] Oral administration of RQ-NPs decreased body weight and fat mass, improved glucose homeostasis, decreased liver inflammatory response and liver damage, enhanced gut integrity, and optimized the microbiota composition of HFD-fed obese mice.

[0334] Furthermore, encapsulated R and Q into lipid nanoparticles (NPs) were made of phospholipids, vitamin E, and surfactant. Obese mice were given the RQ-NPs via oral administration. As compared to free RQ-treated mice, those RQ-NPs-treated mice had significantly lower body weight and fat mass, improved glucose homeostasis, decreased liver inflammatory response and liver damage, enhanced gut integrity, and optimized the microbiota composition of HFD-fed obese mice.

[0335] From these results, it can be seen that biodegradable RQ-NPs increase stability and bioavailability of R and Q, resulting in improved gut integrity, health, glucose homeostasis, and microbiota improvement. Further, RQ-NPs decreased inflammation as well as body weight and fat mass without a loss in lean mass. As such, biocompatible and biodegradable RQ-NPs can be used to combat obesity, diabetes, fatty liver disease, cardiovascular disease, and other metabolic diseases and disorders.

[0336] The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

CLAIMSWhat is claimed is:

1. A nanoparticle comprising: a) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, wherein resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, is present in a concentration range of about 0.1 wt% to about 90 wt%; b) quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, wherein quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, is present in a concentration range of about 0.1 wt% to about 90 wt%; c) at least one lipid, wherein the lipid is present in a concentration range of about 5 wt% to about 99 wt%; d) at least one surfactant, wherein the surfactant is present in a concentration range of about 10 wt% to about 75 wt%; and e) at least one vitamin E, wherein the vitamin E is present in a concentration range of about 5 wt% to about 85 wt%.

2. The nanoparticle of claim 1, wherein the at least one lipid is selected from the group consisting of a fatty acid, wax, sterol, lipid-soluble vitamin, vitamin A, vitamin D, vitamin E, vitamin K, monoglyceride, diglyceride, triglyceride, phospholipid, phosphatidylcholine, L-a- phosphatidylcholine (PC), soy PC, egg PC, phosphatidic acid (PA), phosphatidylethanolamine (PE), lecithin, phosphatidylserine (PS), phosphoinositide, phosphatidylinositol (PI), phosphatidylinositol phosphate (PIP), phosphatidylinositol bisphosphate (PIP2), phosphatidylinositol trisphosphate (PIP3), phosphosphingolipid, sphingolipid, ceramide phosphoryl choline (SPH), ceramide phosphorylethanolamine (Cer-PE), ceramide phosphoryl lipid, and any combination thereof.

3. The nanoparticle of claim 2, wherein the at least one lipid is phosphatidylcholine.

4. The nanoparticle of claim 1, wherein the at least one surfactant is selected from the group consisting of a polyethylene glycol (PEG), functionalized PEG, polyethylene glycol(15)-hydroxy stearate , polyethylene glycol (15)-hydroxystearate containing PEG, polysorbate 80, phospholipids, PEG21- stearate, PEG40-stearate, PEGlOO-stearate, PEG(10-1000)-fatty acid, mustard, lecithin, soy lecithin, egg lecithin, monoglycerides, diglycerides, polysorbates, carrageenan, guar gum, canola oil, polysorbates, sodium dodecyl sulfate, lauryl dimethyl amine oxide, cetyltrimethylammonium bromide (CTAB), polyethoxylated alcohols, polyoxyethylene sorbitan, octoxynol, N, N-dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide (HTAB), polyoxyl 10 lauryl ether, bile salts, sodium deoxy cholate, sodium cholate, polyoxyl castor oil, nonylphenol ethoxylate, cyclodextrins, methylbenzethonium chloride, and any combination thereof.

5. The nanoparticle of claim 4, wherein the at least one surfactant is polyethylene glycol (15)-hydroxystearate , polyethylene glycol (15)-hydroxystearate containing PEG.

6. The nanoparticle of claim 1, wherein the at least one vitamin E is selected from the group consisting of a-tocopherol, a-tocopherol acetate (aTA), a-tocopherol nicotinate, D-a- tocopheryl polyethylene glycol 1000 succinate, P-tocopherol, P-tocopherol acetate, P-tocopherol nicotinate, y-tocopherol, y-tocopherol acetate, y-tocopherol nicotinate, 8-tocopherol, 8-tocopherol acetate, 8-tocopherol nicotinate, a-tocotrienol, a-tocotrienol acetate, a-tocotrienol nicotinate, P- tocotrienol, P-tocotrienol acetate, P-tocotrienol nicotinate, y-tocotrienol, y-tocotrienol acetate, y- tocotrienol nicotinate, 8-tocotrienol, 6-tocotrienol acetate, 8-tocotrienol nicotinate, and any combination thereof.

7. The nanoparticle of claim 6, wherein the at least one vitamin E is aTA.

8. The nanoparticle of claim 1, wherein the nanoparticle further comprises at least one targeting agent.

9. The nanoparticle of claim 8, wherein the at least one targeting agent targets an adipose tissue.

10. The nanoparticle of claim 9, wherein the adipose tissue is a white adipose tissue (WAT), beige adipose tissue, brown adipose tissue (BAT), or any combination thereof.

11. The nanoparticle of claim 10, wherein the targeting agent is selected from the group consisting of proteins, peptides, antibodies, lipids, polysaccharides, nucleic acids, aptamers, and small molecules.

12. The nanoparticle of claim 1, wherein the composition further comprises at least one phytochemical or bioactive compound selected from the group consisting of curcumin, theaflavins, thearubigins, epigallocatechin gallate (EGCG), (+)-catechin, (-)-epicatechin, (-)- epicatechin gallate, (-)-epigallocatechin, (+)-gallocatechin, isorhamnetin, kaempferol, myricetin, apigenin, luteolin, baicalein, chrysin, forskolin, chlorophyll a, chlorophyll b, eriodictyol, hesperetin, naringenin, taxifolin, catechins, luteolin, cyanidin, genistein, daidzein, genistein, glycitein, biochanin A, formononetin, cyanidin, delphinidin, malvidin, pelargonidin, peonidin, petunidin, proanthocyanidins, a-carotene, P-carotene, -cryptoxanthin, lutein, zeaxanthin, lycopene, and any combination thereof.

13. The nanoparticle of claim 1, wherein the nanoparticle further comprises at least one coating agent.

14. The nanoparticle of claim 1, wherein the nanoparticle is a biodegradable nanoparticle.

15. The nanoparticle of any one of claims 1-14, wherein the nanoparticle increases bioavailability of resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

16. The nanoparticle of claim 15, wherein the nanoparticle: a) induces browning of a white adipose tissue (WAT), beige adipose tissue, or any combination thereof; b) increases the level or activity of a brown adipose tissue (BAT), beige adipose tissue, or any combination thereof; c) increases the concentration of cellular cyclic adenosine monophosphate(cAMP); d) increases the level or activity of uncoupling protein 1 (UCP1) or PR domain containing 16 (PRDM16); e) increases the level or activity of a 5' adenosine monophosphate-activated protein kinase (AMPK); f) increases the level or activity of sirtuin 1 (SIRT1); g) increases the level or activity of tight junction protein-related genes; h) increases the level or activity of occluding (OCLN); i) increases the level or activity of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-la); j) decreases the level or activity of a fatty acid synthase (FAS), acetyl-CoA carboxylase (ACC), or a combination thereof; k) decreases the level or activity of NF-KB, tumor necrosis factor alpha (TNF-a), interferon-y, Interleukin 6 (IL-6), IL-4, IL- la, monocyte chemoattractant protein 1 (MCP1), or a combination thereof; l) decreases the level or activity of a phosphodiesterase (PDE); or any combination thereof.

17. The nanoparticle of claim 15, wherein the nanoparticle: a) increases a mitochondrial biogenesis, mitochondrial function, thermogenesis, or a combination thereof; b) increases gut integrity, gut health, or a combination thereof; c) increases insulin sensitivity; d) increases energy expenditure of at least one cell of interest; e) improves metabolic health; f) improves gut microbiota profde; g) improves glucose homeostasis, lipid homeostasis, or a combination thereof; h) decreases fat mass and body weight; i) decreases inflammatory response; j) decrease liver damage; orany combination thereof.

18. The nanoparticle of claim 15, wherein the nanoparticle does not change the level or concentration of lean mass.

19. A composition comprising at least one nanoparticle of claim 15.

20. The composition of claim 19, wherein the composition is selected from the group consisting of an oral formulation, an injectable solution, a solution for transdermal delivery, an injectable hydrogel, a surgically implantable hydrogel, and any combination thereof.

21. A method of increasing the concentration of a cellular cyclic adenosine monophosphate (cAMP) in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of claim 15; or b) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

22. The method of claim 21, wherein the method increases the concentration of a cellular cyclic adenosine monophosphate (cAMP) in an adipose tissue.

23. A method of selectively delivering resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof to an adipose tissue, wherein the method comprises administering: a) at least one nanoparticle of claim 15; or b) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

24. A method of inducing browning of an adipose tissue in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of claim 15; or b) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

25. The method of claim 22, wherein the adipose tissues is a white adipose tissue (WAT), beige adipose tissue, brown adipose tissue (BAT), or any combination thereof.

26. A method of reducing body fat in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of claim 15; or b) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

27. A method of inducing weight loss in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of claim 15; or b) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

28. A method of treating or preventing a metabolic disease or disorder in a subject in need thereof, wherein the method comprises administering to the subject: a) at least one nanoparticle of claim 15; or b) resveratrol, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, quercetin, or a derivative, prodrug, or pharmaceutically acceptable salt thereof, or any combination thereof.

29. The method of claim 28, wherein the metabolic disease or disorder is selected from the group consisting of obesity, obesity-related disease or disorder, diabetes, fatty liver disease, non-alcoholic fatty liver disease, cardiovascular disease, heart disease, stroke, hypertension, idiopathic intracranial hypertension, coronary artery disease, atrial fibrillation, atherosclerosis, venous thromboembolism, obstructive sleep apnea , obesity hypoventilation syndrome, metabolic syndrome, insulin resistance, esophageal adenocarcinoma, gastric cancer, colorectal cancer, hepatocellular carcinoma, cholangiocarcinoma, pancreatic cancer, endometrial carcinoma, ovarian cancer, breast cancer, renal cell carcinoma, multiple myeloma, and any combination thereof.

30. The method of claim 28, wherein the composition is administered via oral administration, injection, or a combination thereof.

31. The method of claim 28, wherein the method further comprises a fecal microbiota transplantation.

32. The method of claim 31, wherein the fecal microbiota transplantation is a human fecal microbiota transplantation (HFT).

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