Uses and methods for promoting increased mitochondrial mass and function

Abstract

Disclosed herein is a method for promoting an increase in mitochondrial mass and function by providing a consumables composition. Some embodiments provided include, for example, administering a compound of formula (I) or a compound of formula (II). Some embodiments provide the composition formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical composition.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 300364, filed January 18, 2022, which is incorporated by reference in its entirety. [Background technology]

[0002] HNF4α is a nuclear receptor transcription factor that controls the expression of downstream genes that are important in multiple aspects of cellular metabolism. The classical view of HNF4α is that its ligand-binding pocket (LBP) is constitutively occupied by fatty acids that play a structural, but not a regulatory, role. However, it has recently been shown that fatty acids in the HNF4α LBP are exchangeable, particularly in cells, in the context of full-length HNF4α. Summary of the Invention

[0003] Disclosed herein is a method for promoting mitochondrial mass and function in a subject in need thereof.In some embodiments, the method comprises administering to a subject in need thereof an oral composition comprising at least one carrier and an effective amount of N-trans-caffeoyltyramine, wherein the subject is eating a high-fat meal.

[0004] Features and advantages of the compositions and methods described herein will become apparent from the following description in conjunction with the accompanying drawings. These drawings illustrate certain aspects of the compositions and methods described in this application and are therefore not to be considered limiting. In the drawings, like reference numbers or symbols typically identify like components unless otherwise indicated by the context. The drawings may not be to scale. [Brief description of the drawings]

[0005] [Figure 1A]Liver sections from mice fed a standard chow (NC), a high fat diet (HFD), or HFD+NCT were immunostained for HNF4α (green nuclear stain) and DAPI (blue nuclear stain). Scale bar = 200 μM. [Figure 1B] Quantification of HNF4α fluorescence intensity from images stained as in FIG. 1A is illustrated. Nonspecific cytoplasmic staining was subtracted from HNF4α nuclear staining and fold changes were calculated relative to NC (NC, N=3; HFD and HFD+NCT, N=12). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). [Figure 1C] Figure 1 illustrates that body weight was measured weekly for 10 weeks. Weight gain was calculated by subtracting baseline body weight at the start of the study (NC N=5, HFD and HFD+NCT, N=15). Values ​​represent mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). [Figure 1D] HFD and HFD+NCT food intake per cage was measured weekly for 10 weeks and demonstrates no difference between the two groups (5 cages for each condition, 3 mice per cage). Values ​​represent mean ± SEM. NS: not significant. [Figure 1E] Figure 1 illustrates fecal TG normalized to fecal dry weight (N=3). Points indicate individual mice. Values ​​represent mean±SEM. NS: not significant. [Figure 1F] Photographs of mice from each treatment group are shown, demonstrating reduced liver size and increased redness, as well as visceral obesity, with NCT resulting in reduced liver size and increased redness. The red box indicates the liver and the arrow indicates the epididymal fat pad. [Figure 1G] Photographs of dissected livers demonstrating reduction in liver size and increased redness with NCT. [Figure 1H]Liver weights were normalized to body weight (NC, N=5; HFD and HFD+NCT, N=15). Points indicate individual mice. Values ​​represent mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). [Figure 1I] Illustrated is a photomicrograph of a liver section stained with Oil Red O. [Figure 1J] Quantification of Oil Red O staining HFD and HFD+NCT values ​​was measured using Image J with consistent thresholding and normalized to NC values ​​to calculate fold change (NC, N=5; HFD and HFD+NCT, N=15). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). [Figure 1K] Liver TG levels were normalized to liver weight (NC, N=5, HFD and HFD+NCT, N=15). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). [Figure 1L] Representative photographs of epididymal fat pads from each group are shown. [Figure 1M] Figure 1 shows epididymal fat pad weight quantified by normalizing to total mouse body weight (NC, N=5, HFD and HFD+NCT, N=15). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). NS: not significant. [Figure 1N] Photographs of mice from each treatment group are shown, demonstrating a reduction in subcutaneous fat (indicated by arrows). [Figure 2A] FIG. 1 illustrates FAO activity in liver lysates prepared in the presence of octanoyl-CoA. Points indicate individual mice (N=6). Values ​​represent the mean±SEM. *p<0.05, **p<0.01 (HFD vs. HFD+NCT). [Figure 2B]Figure 1 illustrates FAO activity prepared in the absence of octanoyl-CoA. Points indicate individual mice (N=6). Values ​​represent mean ± SEM. *p<0.05, **p<0.01 (HFD vs. HFD+NCT). [Figure 2C] FAO activity + octanoyl-CoA - FAO activity - octanoyl-CoA are illustrated. Points represent individual mice (N=6). Values ​​represent the mean ± SEM. NS: not significant. [Figure 2D] Total liver NAD levels are illustrated. Points represent individual mice (N=6). Values ​​represent the mean ± SEM. *p<0.05, **p<0.01 (HFD vs. HFD+NCT). [Figure 3A] Liver sections from NC, HFD, and HFD+NCT mice were immunostained for VDAC-1 (red). DAPI (blue) is for nuclear staining. Scale bar = 200 μM. [Figure 3B] Quantification of hepatic VDAC-1 staining intensity is shown (fold change vs. NC, NC, N=3, HFD and HFD+NCT, N=12). Each point represents an individual mouse. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD v 623 s NC or HFD+NCT). [Figure 3C] Figure 2 illustrates that citrate synthase (CS) activity in liver lysates was reduced by HFD and reversed by NCT ​​(NC, N=3, HFD and HFD+NCT, N=12). Each point represents an individual mouse. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD v 623 s NC or HFD+NCT). [Figure 3D] Western blots for cytochrome C and SDHA are shown. After detection of each protein, the membrane was stained for Ponceau S as a control for protein loading. [Figure 3E]Illustrates quantification of cytochrome C protein expression in mouse liver. Each value was first normalized to Ponceau S staining, then fold change was calculated relative to NC (N=6). Each point represents an individual mouse. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD v 623 s NC or HFD+NCT). [Figure 3F] Illustrates quantification of SDHA protein expression in mouse liver. Each value was first normalized to Ponceau S staining, then fold change was calculated relative to NC (N=6). Each point represents an individual mouse. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD v 623 s NC or HFD+NCT, 0 μM vs. each concentration of NCT in human hepatocytes). NS: not significant. [Figure 3G] Illustrates qPCR analysis in mouse liver for cytochrome C mRNA expression normalized to 18s rRNA (NC, N=3, HFD and HFD+NCT, N=12). Each point represents an individual mouse. Values ​​represent the mean ± SEM. NS: not significant. [Figure 3H] Illustrates qPCR analysis in mouse liver for SDHA mRNA expression normalized to 18s rRNA (NC, N=3, HFD and HFD+NCT, N=12). Each point represents an individual mouse. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD v 623 s NC or HFD+NCT). NS: not significant. [Figure 3I] Illustrates qPCR analysis of cytochrome C mRNA from human primary hepatocytes cultured with the indicated concentrations of NCT (0, 5, 15, 40 μM). Values ​​were normalized to 18s rRNA (N=3). Each point represents an individual human donor. Values ​​represent the mean ± SEM. NS: not significant. [Figure 3J]Illustrated is qPCR analysis of SDHA mRNA from human primary hepatocytes cultured with the indicated concentrations of NCT (0, 5, 15, 40 μM). Values ​​were normalized to 18s rRNA (N=3). Each point represents an individual human donor. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (0 μM vs. each concentration of NCT in human hepatocytes). NS: not significant. [Figure 3K] Illustrates qPCR analysis in mouse liver for mitochondrial DNA (ND1) expression normalized to HK2 (N=4). Each point represents an individual mouse. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD v 623 s NC or HFD+NCT). [Figure 3L] Illustrates qPCR analysis in mouse liver for mitochondrial DNA (16s) expression normalized to HK2 (N=4). Each point represents an individual mouse. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD v 623 s NC or HFD+NCT). [Figure 3M] FIG. 1 illustrates qPCR analysis in mouse liver for HSP60 mRNA expression normalized to 18s rRNA (NC, N=3, HFD and HFD+NCT, N=12). Each point represents an individual mouse. Values ​​represent the mean±SEM. [Figure 3N] Illustrates qPCR analysis in mouse liver for PPARγ mRNA expression normalized to 18s rRNA (NC, N=3, HFD and HFD+NCT, N=12). Each point represents an individual mouse. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD v 623 s NC or HFD+NCT). [Figure 4A] Representative photographs of Western blot analysis of PPARGC1A expression and Ponceau S in mouse liver are shown. [Figure 4B]Figure 1 illustrates PPARGC1A protein expression (fold change relative to NC) in mouse liver normalized with Ponceau S (N=6). Points indicate individual mice. Values ​​represent mean ± SEM. *p<0.05, **p<0.01 (HFD vs. NC or HFD+NCT). [Figure 4C] Illustrates qPCR analysis of PPARGC1A mRNA levels in mouse liver normalized to 18s rRNA (NC, N=3; HFD and HFD+NCT, N=12). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01 (HFD vs. NC or HFD+NCT). NS: not significant. [Figure 4D] Illustrates qPCR analysis of Sirtuin 1 mRNA levels in mouse liver normalized to 18s rRNA (NC, N=3; HFD and HFD+NCT, N=12). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01 (HFD vs. NC or HFD+NCT). [Figure 4E] Illustrates qPCR analysis of Sirtuin 3 mRNA levels in mouse liver normalized to 18s rRNA (NC, N=3; HFD and HFD+NCT, N=12). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01 (HFD vs. NC or HFD+NCT). [Figure 4F] Illustrates qPCR analysis of 18s rRNA-normalized PPARGC1A mRNA levels in human primary hepatocytes (N=3). Points indicate human donors. Values ​​represent mean±SEM. *p<0.05, **p<0.01 (0 μM vs. each concentration of NCT in human hepatocytes). NS: not significant. [Figure 4G] Illustrates qPCR analysis of 18s rRNA-normalized Sirtuin 1 mRNA levels in human primary hepatocytes (N=3). Points indicate human donors. Values ​​represent mean±SEM. *p<0.05, **p<0.01 (0 μM vs. each concentration of NCT in human hepatocytes). NS: not significant. [Figure 4H]Illustrates qPCR analysis of 18s rRNA-normalized Sirtuin 3 mRNA levels in human primary hepatocytes (N=3). Points indicate human donors. Values ​​represent mean±SEM. *p<0.05, **p<0.01 (0 μM vs. each concentration of NCT in human hepatocytes). NS: not significant. [Figure 5A] Illustrates qPCR analysis in mouse liver of IL-6 mRNA levels normalized to 18s rRNA (NC, N=3; HFD and HFD+NCT, N=12). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). NS: not significant. [Figure 5B] Illustrated is qPCR analysis in mouse liver of TNFα mRNA levels normalized to 18s rRNA (NC, N=3; HFD and HFD+NCT, N=12). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). NS: not significant. [Figure 5C] Illustrated is qPCR analysis in mouse liver of IL-1β mRNA levels normalized to 18s rRNA (NC, N=3, HFD and HFD+NCT, N=12). Points indicate individual mice. Values ​​represent mean±SEM. NS: not significant. [Figure 5D] Figure 1 illustrates ELISA analysis of IL-6 secreted into the medium of human primary hepatocytes cultured with palmitate and DMSO (vehicle control) or different concentrations of NCT (N=3). Points indicate individual human donors. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (0 μM vs. each concentration of NCT in human hepatocytes). NS: not significant. [Figure 5E] Figure 1 illustrates qPCR analysis of IL-6 mRNA levels in primary human hepatocytes. Normalization was performed against 18s rRNA (N=3). Points indicate individual human donors. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (0 μM vs. each concentration of NCT in human hepatocytes). NS: not significant. [Figure 5F] Figure 1 illustrates qPCR analysis of TNFα mRNA levels in primary human hepatocytes. Normalization was performed against 18s rRNA (N=3). Points indicate individual human donors. Values ​​represent the mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (0 μM vs. each concentration of NCT in human hepatocytes). NS: not significant. [Figure 5G] Illustrated is an in vivo nitric oxide (NO) analysis. NO expression normalized to liver weight in vivo (NC, N=3; HFD and HFD+NCT, N=4). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). [Figure 5H] Illustrates in vitro nitric oxide (NO) analysis. NO expression normalized to protein in T6PNE cells in vitro (N=3). Points indicate human donors. Values ​​represent mean ± SEM. *p<0.05, **p<0.01, ***p<0.001 (0 μM vs. each concentration of NCT in human T6PNE cells). [Figure 5I] FIG. 1 illustrates that blood alanine aminotransferase (ALT) levels were reduced in HFD+NCT mice compared to HFD mice (NC, N=3; HFD, N=12; HFD+NCT, N=9). Points indicate individual mice. Values ​​represent mean±SEM. *p<0.05, **p<0.01, ***p<0.001 (HFD vs. NC or HFD+NCT). [Figure 6A] Figure 1 illustrates the precipitation of NCT at the subcutaneous injection site of a representative mouse. The red arrow indicates the precipitation of the compound. [Figure 6B] Representative photographs of dissected subcutaneous NCT and DMSO injected mice are shown. [Figure 6C] Photographs of the dissected livers from representative mice (N=6) are shown. There is no difference in the color of the liver. [Figure 6D] Liver weights (normalized to body weight) were measured (N=6). Points indicate individual mice, NS=not significant. Values ​​represent the mean±SEM. [Figure 6E] Representative photographs of dissected methylcellulose and NCT treated mice (N=5) are shown. [Figure 6F] Representative pictures of dissected stomachs of methylcellulose (MC) and NCT treated mice (N=5) are shown. Right: Intact stomach. Left: Open stomach, red arrows indicate NCT deposits in the stomach. [Figure 6G] Representative photographs of dissected livers are shown. There is no difference in the color of the liver. [Figure 6H] Representative pictures of Oil Red O staining on liver sections are shown. Scale bar = 200 μM. [Figure 6I] Figure 1 illustrates NCT compound levels measured in serum and feces (N=4). Points represent individual mice. Values ​​represent the mean ± SEM. [Figure 7A] Immunostaining for cleaved caspase 3 (red) and DAPI (blue) is shown. Scale bar = 200 μM. [Figure 7B] Total DAPI counts per well are depicted. Values ​​represent mean ± SEM. NS: not significant. [Figure 7C] Quantification of cleaved caspase 3 positive cells normalized to cell number measured with DAPI is shown. Values ​​represent mean ± SEM. NS: not significant. [Figure 7D] Immunostaining of cleaved caspase 3 (red), HNF4α (green), and DAPI (blue) in mouse liver is shown. Scale bar = 200 μM. [Figure 8A] FIGURE 2 illustrates quantification of TG levels in serum samples collected from mice fed HFD and NCT-treated HFD diets after 10 weeks (N=6). Points indicate individual mice, NS=not significant. Values ​​represent mean±SEM. [Figure 8B] Free fatty acids were quantified in serum samples from these mice (N=15). Points indicate individual mice, NS=not significant. Values ​​represent the mean±SEM. [Figure 8C]ALP levels were quantified in blood samples from these mice (N=9). Points indicate individual mice, NS=not significant. Values ​​represent the mean±SEM. [Figure 9A] Figure 1 illustrates qPCR analysis in mouse liver of Parp1 mRNA expression normalized at 18s (NC, N=3, HFD and NCT, N=12). Points indicate individual mice. Values ​​represent mean ± SEM. NS: not significant. [Figure 9B] Figure 1 illustrates qPCR analysis in mouse liver of normalized Parp2 mRNA expression at 18s (NC, N=3, HFD and NCT, N=12). Points indicate individual mice. Values ​​represent mean ± SEM. NS: not significant. [Figure 10] A table is illustrated describing the NCT doses, as well as the dietary treatments and duration of post-treatment monitoring. [Figure 11] Illustrates the liver profile panel and hematological analysis tables. [Figure 12] Illustrates the liver profile panel and hematological analysis tables. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0006] The present disclosure provides, inter alia, the discovery of potent HNF4α agonists and their use to reveal a previously unknown pathway by which HNF4α controls the level of fat storage in the liver. Without wishing to be bound by theory, it is believed that this involves the induction of lipophagy by dihydroceramide, the synthesis and secretion of which is controlled by genes induced by HNF4α. HNF4α activators are N-trans caffeoyl tyramine (NCT) and N-trans feruloyl tyramine (NFT), which are structurally related to the known drugs alverine and benfluorex, which are weak HNF4α activators. In the in vitro studies described herein, NCT and NFT induced fat clearance from palmitate-loaded cells.

[0007] In aspects, provided herein are compounds and compositions containing hydroxycinnamic acid amide-containing tyramines.Some embodiments provided herein provide compounds and compositions for use in methods of promoting increased mitochondrial mass and function.

[0008] composition In some aspects, the disclosure provided herein provides plant-derived aromatic metabolites having one or more acidic hydroxyl groups attached to aromatic arenes and their use in metabolic regulation. In one embodiment, the plant-derived aromatic metabolites are structural analogs of compound 1. [ka]

[0009] In particular, the disclosure encompasses compounds of formula (I) or an isomer, salt, homodimer, heterodimer, or conjugate thereof: [ka]

[0010] In some embodiments, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , and R 9 are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C 1-6 Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Heterocyclyl, optionally substituted -(O)C 4-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-12 Heteroaryl and optionally substituted -(O)C 1-6 Alkyl C 1-12 Heteroaryl is selected from the group consisting of aryl, hetero ...

[0011] In some embodiments, R 1 , R 2 , R 3 , and R 8 are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C 1-6 Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Heterocyclyl, optionally substituted -(O)C 4-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-12 Heteroaryl and optionally substituted -(O)C 1-6Alkyl C 1-12 heteroaryl; R 4 , R 5 , R 6 , R 7 , and R 9 are each independently hydrogen, deuterium, hydroxyl, or halogen.

[0012] In some embodiments, R 1 , R 2 , and R 8 are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C 1-6 Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Heterocyclyl, optionally substituted -(O)C 4-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-12 Heteroaryl and optionally substituted -(O)C 1-6 Alkyl C 1-12 heteroaryl; R 3 , R 4 , R 5 , R 6 , R 7 , and R 9 are each independently hydrogen, deuterium, hydroxyl, or halogen.

[0013] In some embodiments, the dashed bond is present or absent.

[0014] In some embodiments, X is CH 2 Or O.

[0015] In some embodiments, Z is CHR a , N.R. a , or O.

[0016] In some embodiments, R a is hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amide, optionally substituted N-amide, optionally substituted ester, optionally substituted -(O)C 1-6 Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Heterocyclyl, optionally substituted -(O)C 4-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-12 Heteroaryl and optionally substituted -(O)C 1-6 Alkyl C 1-12 Heteroaryl is selected from the group consisting of aryl, hetero ...

[0017] In some embodiments, the compound of formula (I) is provided as a pharma- ceutically acceptable salt or solvate thereof.

[0018] In some embodiments, the compound of formula (I) is selected from the group consisting of (E)-3-(3,4-dihydroxyphenyl)-N-(4-ethoxyphenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(2-methoxyethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(2-(methylsulfonyl)ethoxy)phenethyl)acrylamide, (E)-2-(4-(2-(3-(3,4-dihydroxyphenyl)acrylamido)ethyl)phenoxy)acetic acid, ethyl (E)-2-(4-( 2-(3-(3,4-dihydroxyphenyl)acrylamido)ethyl)phenoxy)acetate, (E)-N-(4-(cyclopropylmethoxy)phenethyl)-3-(3,4-dihydroxyphenyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(3,3,3-trifluoropropoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((tetrahydro-2H-pyran-4-yl)methoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((tetrahydro-2H-pyran-4-yl)methoxy)phenethyl)acrylamide (E)-N-(4-(cyanomethoxy)phenethyl)-3-(3,4-dihydroxyphenyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(pyridin-3-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(pyridin-2-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(2-(dimethylamino)ethoxy)-1,3-dihydroxyphenyl)-N-(4-(pyridin-3-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-isobutoxyphenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(pyridin-4-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((4-methoxybenzyl)oxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(oxetan-3-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((tetrahydro-2H-pyran-2-yl)methoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((tetrahydrofuran-2-yl)methoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(thiophen-2-yloxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(3,3-dimethylbutoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(2-hydroxyethoxy)phenethyl)acrylamide, (E)-N-(4-((1H-tetrazol-5-yl)methoxy)phenethyl)-3-(3,4-dihydroxyphenyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((1-methylpyrrolidin-2-yl)methoxy)phenethyl)acrylamide, (E)-2-hydroxy-5-(3-((4-hydroxyphenethyl)amino)-3-oxoprop-1-en-1-yl)phenyl hydrogen carbonate, (E)-3-(4-hydroxy-3-(pyridin-4-yloxy)phenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-3-(4-hydroxy-3-isobutoxyphenyl)-N-(4-hydroxyphenethyl)acryl acrylamide, (E)-3-(3-(4-fluorophenoxy)-4-hydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-3-(3-(cyanomethoxy)-4-hydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-2-(2-hydroxy-4-(3-((4-hydroxyphenethyl)amino)-3-oxoprop-1-en-1-yl)phenoxy)acetic acid, (E)-3-(3-hydroxy-4-(pyridin-4-ylmethoxy)phenyl)-N-(4-hydroxyphenyl)- (E)-3-(4-((4-fluorobenzyl)oxy)-3-hydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-3-(3-hydroxy-4-isobutoxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-3-(4-(cyanomethoxy)-3-hydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-N-(3-(3,4-dihydroxyphenyl)acryloyl)-N-(4-hydroxyphenyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-hydroxyphenethyl)-N-(pyridin-4-ylmethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-hydroxyphenethyl)-N-isobutylacrylamide, (E)-N-(cyanomethyl)-3-(3,4-dihydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, 3-(3,4-dihydroxyphenyl)-N-(4-hydroxyphenethyl)propanamide, 3-(3,4-dihydroxyphenyl)-N-(4-(methylsulfonamido)phenethyl)propanamide, or a pharmaceutical salt, solvate, or combination thereof.

[0019] In some embodiments, the disclosure includes a compound of formula (II). [ka]

[0020] In some embodiments, R 1 , R 2 , R 3 , and R 4 are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C 1-6 Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Heterocyclyl, optionally substituted -(O)C 4-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-12 Heteroaryl and optionally substituted -(O)C 1-6 Alkyl C 1-12 Heteroaryl is selected from the group consisting of aryl, hetero ...

[0021] In some embodiments, the dashed bond is present or absent.

[0022] In some embodiments, Z is CHR a , N.R. a , or O.

[0023] In some embodiments, R a is hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amide, optionally substituted N-amide, optionally substituted ester, optionally substituted -(O)C 1-6 Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Heterocyclyl, optionally substituted -(O)C 4-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-12 Heteroaryl and optionally substituted -(O)C 1-6 Alkyl C 1-12 Heteroaryl is selected from the group consisting of aryl, hetero ...

[0024] In some embodiments, the compound of formula (II) is (E)-3-(3,4-dihydroxyphenyl)-N-(4-ethoxyphenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(2-methoxyethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(2-(methylsulfonyl)ethoxy)phenethyl)acrylamide, (E)-2-(4-(2-(3-(3,4-dihydroxyphenyl)acrylamido)ethyl)phenoxy)acetic acid, ethyl (E)-2-(4- (2-(3-(3,4-dihydroxyphenyl)acrylamide)ethyl)phenoxy)acetate, (E)-N-(4-(cyclopropylmethoxy)phenethyl)-3-(3,4-dihydroxyphenyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(3,3,3-trifluoropropoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((tetrahydro-2H-pyran-4-yl)methoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((tetrahydro-2H-pyran-4-yl)methoxy)phenethyl)acrylamide, (E)-N-(4-(4-fluorobenzyl)oxy)phenethyl)acrylamide, (E)-N-(4-(cyanomethoxy)phenethyl)-3-(3,4-dihydroxyphenyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(pyridin-3-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(pyridin-2-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(2-(dimethyl)ethoxy) (E)-3-(3,4-dihydroxyphenyl)-N-(4-isobutoxyphenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(pyridin-4-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((4-methoxybenzyl)oxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(oxetan-3-ylmethoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((tetrahydro-2H-pyran-2-yl)methoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((tetrahydrofuran-2-yl)methoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(thiophen-2-yloxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(3,3-dimethylbutoxy)phenethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-(2-hydroxyethoxy)phenethyl)acrylamide, (E)-N-(4-((1H-tetrazol-5-yl)methoxy)phenethyl)-3-(3,4-dihydroxyphenyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-((1-methylpyrrolidin-2-yl)methoxy)phenethyl)acrylamide, (E)-2-hydroxy-5-(3-((4-hydroxyphenethyl)amino)-3-oxoprop-1-en-1-yl)phenyl hydrogen carbonate, (E)-3-(4-hydroxy-3-(pyridin-4-yloxy)phenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-3-(4-hydroxy-3-isobutoxyphenyl)-N-(4-hydroxyphenethyl)acryl acrylamide, (E)-3-(3-(4-fluorophenoxy)-4-hydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-3-(3-(cyanomethoxy)-4-hydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-2-(2-hydroxy-4-(3-((4-hydroxyphenethyl)amino)-3-oxoprop-1-en-1-yl)phenoxy)acetic acid, (E)-3-(3-hydroxy-4-(pyridin-4-ylmethoxy)phenyl)-N-(4-hydroxyphenyl)- (E)-3-(4-((4-fluorobenzyl)oxy)-3-hydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-3-(3-hydroxy-4-isobutoxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-3-(4-(cyanomethoxy)-3-hydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, (E)-N-(3-(3,4-dihydroxyphenyl)acryloyl)-N-(4-hydroxyphenyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-hydroxyphenethyl)-N-(pyridin-4-ylmethyl)acrylamide, (E)-3-(3,4-dihydroxyphenyl)-N-(4-hydroxyphenethyl)-N-isobutylacrylamide, (E)-N-(cyanomethyl)-3-(3,4-dihydroxyphenyl)-N-(4-hydroxyphenethyl)acrylamide, 3-(3,4-dihydroxyphenyl)-N-(4-hydroxyphenethyl)propanamide, 3-(3,4-dihydroxyphenyl)-N-(4-(methylsulfonamido)phenethyl)propanamide, or a pharmaceutical salt, solvate, or combination thereof.

[0025] In some embodiments, the compound of formula (II) is provided as a pharma- ceutically acceptable salt or solvate thereof.

[0026] In some embodiments, the disclosure includes a compound of formula (III). [ka] In some embodiments, R 3 and R 4 are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C 1-6 Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 1-6 Alkyl C 2-12 Heterocyclyl, optionally substituted -(O)C 5-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-12 Heteroaryl and optionally substituted -(O)C 1-6 Alkyl C 1-12 Heteroaryl is selected from the group consisting of aryl, hetero ...

[0027] In some embodiments, each independently selected dashed bond is present or absent.

[0028] In some embodiments, Z is CHR a , N.R. a , or O.

[0029] In some embodiments, R a is hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amide, optionally substituted N-amide, optionally substituted ester, optionally substituted -(O)C 1-6 Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Heteroaryl is selected from the group consisting of aryl, hetero ...

[0030] In some embodiments, Q a , Q b , Q c , Q d each independently represents a bond, CHR a , N.R. a , C═O, and —O—.

[0031] In some embodiments, R a is hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amide, optionally substituted N-amide, optionally substituted ester, optionally substituted -(O)C 1-6Alkyl, optionally substituted -(O)C 1-6 Alkenyl, optionally substituted -(O)C 1-6 Alkynyl, optionally substituted -(O)C 4-12 Cycloalkyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Cycloalkyl, optionally substituted -(O)C 4-12 Heterocyclyl, optionally substituted -(O)C 1-6 Alkyl C 4-12 Heterocyclyl, optionally substituted -(O)C 4-12 Aryl, optionally substituted -(O)C 1-6 Alkyl C 5-12 Aryl, optionally substituted -(O)C 1-12 Heteroaryl and optionally substituted -(O)C 1-6 Alkyl C 1-12 Heteroaryl is selected from the group consisting of aryl, hetero ...

[0032] In some embodiments, Q c , Q d In some embodiments, Q is absent. d is absent.

[0033] In some embodiments, n is 1, 2, 3, or 4.

[0034] In some embodiments, the compound of formula (II) is provided as a pharma- ceutically acceptable salt or solvate thereof.

[0035] "Isomers" refers in particular to optical isomers (e.g., essentially pure enantiomers, essentially pure diastereomers, and mixtures thereof), as well as conformational isomers (i.e., isomers that differ only in the angle of at least one chemical bond), positional isomers (especially tautomers), and geometric isomers (e.g., cis-trans isomers).

[0036] In certain embodiments, the compound of formula (I) or formula (II) is selected from: [ka]

[0037] A salt of a compound of the present disclosure refers to a compound that possesses the desired pharmacological activity of the parent compound and includes (1) an acid addition salt formed with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, or an acid addition salt formed with an acid such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzene ... acid addition salts formed with organic acids such as benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, 4-toluenesulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylbutyric acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid; or (2) salts formed when an acidic proton present in the parent compound is replaced.

[0038] As known in the art, homodimers are molecules that are composed of two identical tyramines that contain hydroxycinnamic acid amide subunits. In comparison, heterodimers are molecules that are composed of two different tyramines that contain hydroxycinnamic acid amide subunits. Examples of homodimers in the present disclosure include, but are not limited to, cross-linked N-trans-feruloyltyramine dimers, cross-linked N-trans-caffeoyltyramine dimers, and cross-linked p-coumaroyltyramine dimers. See, for example, King & Calhoun (2005) Phytochemistry 66(20):2468-73, which teaches the isolation of cross-linked N-trans-feruloyltyramine dimers from common scab lesions of potato.

[0039] When a group is described as "optionally substituted," the group may be unsubstituted or substituted with one or more of the indicated substituents. Similarly, when a group is described as "unsubstituted or substituted," if substituted, the substituents may be selected from one or more of the indicated substituents. When no substituents are indicated, this means that the indicated "optionally substituted" or "substituted" group is selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl, aralkyl, heteroaralkyl, (heteroalicyclyl)alkyl, hydroxy, protected hydroxyl, alkoxy, aryloxy, acyl, mercapto, alkylthio, arylthio, cyano, halogen, thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarboxyl, N-thiocarbam ... It means that they may be individually and independently substituted with one or more group(s) individually and independently selected from propyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido, C-carboxy, protected C-carboxy, O-carboxy, isocyanate, thiocyanate, isothiocyanate, nitro, silyl, sulfenyl, sulfinyl, sulfonyl, haloalkyl, haloalkoxy, trihalomethanesulfonyl, trihalomethanesulfonamido, amino, mono-substituted amino groups, and di-substituted amino groups, and protected derivatives thereof.

[0040] For groups in this specification, the subscripts in parentheses below further define the groups as follows: n ) defines the exact number of carbon atoms (n) in the group. For example, "C 1 -C 6 "Alkyl" refers to an alkyl group having 1 to 6 carbon atoms (eg, 1, 2, 3, 4, 5, or 6, or any range derivable therein (eg, 3 to 6 carbon atoms)).

[0041] In addition to isomers, salts, homodimers, heterodimers, and conjugates, hydroxycinnamic acid amide-containing tyramines can also be glycosylated. Hydroxycinnamic acid amide-containing glycosylated tyramines can be produced by transglycosylating hydroxycinnamic acid amide-containing tyramines to add, for example, 1, 2, 3, 4, 5, or more than 5 glucose units to the hydroxycinnamic acid amide-containing tyramine. Transglycosylation can be carried out with any suitable enzyme, including, but not limited to, pullulanase and isomaltase (Lobov, et al. (1991) Agric. Biol. Chem. 55:2959-2965), -galactosidase (Kitahata, et al. (1989) Agric. Biol. Chem. 53:2923-2928), dextrin saccharase (Yamamoto, et al. (1994) Biosci. Biotech. Biochem. 58:1657-1661), or cyclodextrin gluconotransferase, with pullulan, maltose, lactose, partially hydrolyzed starch, and maltodextrin as donors.

[0042] As used herein, "alkyl" refers to a straight or branched hydrocarbon chain that includes a fully saturated (no double or triple bonds) hydrocarbon group. The alkyl group may have 1-20 carbon atoms (each occurrence herein, a numerical range such as "1-20" refers to each integer within the given range, e.g., "1-20 carbon atoms" means that the alkyl group may contain up to 20 carbon atoms and may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., but this definition also encompasses occurrences of the term "alkyl" where no numerical range is specified). The alkyl group may also be a medium sized alkyl having 1-10 carbon atoms. The alkyl group may also be a lower alkyl having 1-6 carbon atoms. The alkyl group of the compound may be represented as "C1-C4 alkyl" or a similar designation. By way of example only, "C1-C4 alkyl" indicates that there are 1-4 carbon atoms in the alkyl chain, i.e., the alkyl chain is selected from methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and t-butyl. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tertiary butyl, pentyl, and hexyl. Alkyl groups can be substituted or unsubstituted.

[0043] As used herein, the term "halogen atom" or "halogen" means any one of the radiostable atoms in column 7 of the periodic table of the elements, such as chloro (Cl), fluoro (F), bromo (Br), and iodo (I) groups.

[0044] In any of the groups described herein, an available hydrogen may be replaced by alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, alkylaryl, heteroaralkyl, heteroarylalkenyl, heteroarylalkynyl, alkylheteroaryl, hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, alkoxyalkoxy, alkoxycarbonyl, acyl, halo, nitro, aryloxycarbonyl, cyano, carboxy, aralkoxycarbonyl, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl, alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio, cycloalkyl, or heterocyclyl.

[0045] Any undefined valency on an atom of a structure shown in this application implicitly represents a hydrogen atom bonded to the atom.

[0046] As used herein, "alkenyl" refers to an alkyl group, as defined herein, containing one or more double bonds in the straight or branched hydrocarbon chain. An alkenyl group can be unsubstituted or substituted.

[0047] As used herein, "alkynyl" refers to an alkyl group, as defined herein, that contains one or more triple bonds in the straight or branched hydrocarbon chain. An alkynyl group can be unsubstituted or substituted.

[0048] As used herein, "cycloalkyl" refers to a fully saturated (no double or triple bonds) monocyclic or polycyclic hydrocarbon ring system. When composed of two or more rings, the rings may be linked together in a fused fashion. Cycloalkyl groups can contain 3 to 10 atoms in the ring(s), or 3 to 8 atoms in the ring(s). Cycloalkyl groups can be unsubstituted or substituted. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0049] As used herein, "aryl" refers to a carbocyclic (all carbon) monocyclic or polycyclic aromatic ring system (e.g., including fused, bridged, or spiro ring systems in which two carbocyclic rings share a chemical bond, e.g., one or more aryl rings with one or more aryl or non-aryl rings) having a fully delocalized pi-electron system throughout at least one of the rings. The number of carbon atoms in an aryl group can vary. For example, an aryl group can be any of C 6 -C 14 Aryl group, C 6 -C 10 Aryl group, or C 6 It may be an aryl group. Examples of aryl groups include, but are not limited to, benzene, naphthalene, and azulene. The aryl group may be substituted or unsubstituted.

[0050] As used herein, "heterocyclyl" refers to a monocyclic or polycyclic ring system containing at least one heteroatom (e.g., O, N, S). Such systems may be unsaturated, may contain some unsaturation, or may contain some aromatic moieties or may be entirely aromatic. Heterocyclyl groups may contain from 3 to 30 atoms. Heterocyclyl groups may be unsubstituted or substituted.

[0051] In certain embodiments, R 1 is present and represents a hydroxy group at the para position, R 2 is a hydroxy group or a lower alkoxy group at the meta position. In certain embodiments, the tyramine containing hydroxycinnamic acid amide having the structure of formula (I) is in the trans configuration.

[0052] As used herein, "heteroaryl" refers to a monocyclic or polycyclic aromatic ring system (a ring system having at least one ring with a fully delocalized π-electron system) containing one or more heteroatoms, i.e., elements other than carbon, including but not limited to nitrogen, oxygen, and sulfur, and at least one aromatic ring. The number of atoms in the ring(s) of a heteroaryl group can vary. For example, a heteroaryl group can contain 4 to 14 atoms in the ring(s), 5 to 10 atoms in the ring(s), or 5 to 6 atoms in the ring(s). Furthermore, the term "heteroaryl" includes fused ring systems in which two rings share at least one chemical bond, such as at least one aryl ring and at least one heteroaryl ring, or at least two heteroaryl rings. Examples of heteroaryl rings include, but are not limited to, furan, furazan, thiophene, benzothiophene, phthalazine, pyrrole, oxazole, benzoxazole, 1,2,3-oxadiazole, 1,2,4-oxadiazole, thiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole, benzothiazole, imidazole, benzimidazole, indole, indazole, pyrazole, benzopyrazole, isoxazole, benzoisoxazole, isothiazole, triazole, benzotriazole, thiadiazole, tetrazole, pyridine, pyridazine, pyrimidine, pyrazine, purine, pteridine, quinoline, isoquinoline, quinazoline, quinoxaline, cinnoline, and triazine. Heteroaryl groups can be substituted or unsubstituted.

[0053] As used herein, the term "amino" refers to -NH 2 Refers to the base.

[0054] As used herein, the term "hydroxy" refers to an --OH group.

[0055] A "cyano" group refers to a "-CN" group.

[0056] A "carbonyl" group refers to a C=O group.

[0057] A "C-amide" group is R A and R B is independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl, aralkyl, or (heteroalicyclyl)alkyl, as defined above; A R B ) group. C-amides can be substituted or unsubstituted.

[0058] An "N-amido" group is one in which R and R A is "RC(═O)N(R )", which may be independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl, aralkyl, or (heteroalicyclyl)alkyl, as defined above. A N-amides refer to an "N-amide" group. N-amides can be substituted or unsubstituted.

[0059] The "urea" group is R A and R B is independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclyl, aralkyl, or (heteroalicyclyl)alkyl, as defined above. A R B )-C(=O)-N(R A R B )-" group. The urea group can be substituted or unsubstituted.

[0060] As used herein, the term "pharmaceutical acceptable salt" is a broad term and is given its usual and customary meaning to those skilled in the art (not limited to any special or customized meaning), and refers to, but is not limited to, a salt of a compound that does not cause significant irritation to the organism to which it is administered and does not abolish the biological activity and properties of the compound. In some embodiments, the salt is an acid addition salt of the compound. Pharmaceutical salts can be obtained by reacting a compound with an inorganic acid, such as hydrohalic acid (e.g., hydrochloric acid or hydrobromic acid), sulfuric acid, nitric acid, and phosphoric acid. Pharmaceutical salts can also be obtained by reacting the compounds with organic acids such as aliphatic or aromatic carboxylic or sulfonic acids, e.g., formic acid, acetic acid (AcOH), propionic acid, glycolic acid, pyruvic acid, malonic acid, maleic acid, fumaric acid, trifluoroacetic acid (TFA), benzoic acid, cinnamic acid, mandelic acid, succinic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, nicotinic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, muconic acid, butyric acid, phenylacetic acid, phenylbutyric acid, valproic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, or naphthalenesulfonic acid. Pharmaceutical salts can also be prepared by reacting a compound with a base to form a salt, for example, an ammonium salt, an alkali metal salt (e.g., lithium salt, sodium salt, or potassium salt), an alkaline earth metal salt (e.g., calcium salt, magnesium salt, or aluminum salt), an organic base (e.g., dicyclohexylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, C 1 -C 7 These can be obtained by forming salts with alkylamines, cyclohexylamine, dicyclohexylamine, triethanolamine, ethylenediamine, ethanolamine, diethanolamine, triethanolamine, tromethamine), and with amino acids (e.g., arginine and lysine), or with inorganic bases (e.g., aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like).

[0061] In any compound described herein having one or more chiral centers, unless the absolute configuration is explicitly indicated, it is understood that each center may be independently in the R or S configuration, or a mixture thereof. Thus, the compounds provided herein may be enantiomerically pure, enantiomerically enriched, or a stereoisomeric mixture, including all diastereomeric and enantiomeric forms. Additionally, in any compound described herein having one or more double bonds that produce geometric isomers that may be defined as E or Z, it is understood that each double bond may be independently E or Z, or a mixture thereof. Stereoisomers may be obtained, if desired, by methods such as stereoselective synthesis and / or separation of stereoisomers by chiral chromatographic columns.

[0062] Likewise, it should be understood that in any compound described, all tautomeric forms are intended to be included.

[0063] It is understood that the compounds described herein may be isotopically or by other means, including but not limited to the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels. Substitution with an isotope, such as deuterium, may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements. Each chemical element depicted in a compound structure may include any isotope of that element. For example, in a compound structure, a hydrogen atom may be explicitly disclosed or understood as being present in the compound. At any position of a compound where a hydrogen atom may be present, the hydrogen atom may be any isotope of hydrogen, including but not limited to hydrogen-1 (protium), hydrogen-2 (deuterium), and hydrogen-3 (tritium). Thus, reference to a compound herein encompasses all possible isotopic forms, unless the context explicitly indicates otherwise.

[0064] It is understood that the compounds described herein may be isotopically or by other means, including but not limited to the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels. Substitution with an isotope, such as deuterium, may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements. Each chemical element depicted in a compound structure may include any isotope of that element. For example, in a compound structure, a hydrogen atom may be explicitly disclosed or understood as being present in the compound. At any position of a compound where a hydrogen atom may be present, the hydrogen atom may be any isotope of hydrogen, including but not limited to hydrogen-1 (protium), hydrogen-2 (deuterium), and hydrogen-3 (tritium). Thus, reference to a compound herein encompasses all possible isotopic forms, unless the context explicitly indicates otherwise.

[0065] It is understood that the methods and formulations described herein include the use of crystalline forms, amorphous phases, and / or pharma- ceutically acceptable salts, solvates, hydrates, and conformers of the compounds of some embodiments, as well as metabolites and active metabolites of these compounds with the same type of activity. Conformers are structures that are stereoisomeric. Conformers are the phenomenon of molecules that have the same structural formula but have different conformations of atoms about a rotational bond (conformers). In certain embodiments, the compounds described herein exist in solvated forms with pharma- ceutically acceptable solvents such as water, ethanol, etc. In other embodiments, the compounds described herein exist in unsolvated forms. Solvates contain either stoichiometric or non-stoichiometric amounts of a solvent and can be formed during the process of crystallization with pharma- ceutically acceptable solvents such as water, ethanol, etc. They form hydrates when the solvent is water, or alcoholates when the solvent is alcohol. In addition, the compounds provided herein can exist in unsolvated and solvated forms. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the compounds and methods provided herein. Other forms in which the compounds of some embodiments may be provided include amorphous, milled, and nanoparticulate forms.

[0066] Similarly, it is understood that the compounds described herein, such as the compounds of some embodiments, include the compounds in any of the forms described herein (e.g., pharma- ceutically acceptable salts, prodrugs, crystalline forms, amorphous forms, solvated forms, enantiomeric forms, tautomeric forms, etc.).

[0067] formulation The substantially pure compound or extract containing the disclosed compound can be combined with a carrier and provided in any suitable form for ingestion by or administration to a subject.In this regard, the compound or extract is added to a consumable product as an exogenous ingredient or additive.Suitable consumable product forms include, but are not limited to, dietary supplements, food ingredients or additives, medical foods, nutraceuticals, or pharmaceutical compositions.In some embodiments, the compound or extract is provided in either liquid or powder form.

[0068] A food ingredient or additive is an edible substance intended, directly or indirectly, to become a component of any food or to otherwise affect its properties (including any substance intended for use in the manufacture, production, packaging, processing, preparation, treatment, packaging, transport, or holding of a food). Foods, particularly functional foods, are foods that have been fortified or enriched during processing to contain additional supplemental nutrients and / or beneficial ingredients. Foods according to the present disclosure can be in the form of, for example, butter, margarine, sweet or savory spreads, condiments, biscuits, health bars, bread, cakes, cereals, candies, confectionery, soups, milk, yogurt or fermented dairy products, cheese, juice-based and vegetable-based beverages, fermented beverages, shakes, flavored waters, teas, oils, or any other suitable food. In some embodiments, the food is a whole food in which the concentration of the compound has been enriched to a level that provides an effective amount of the compound by certain post-harvest and food manufacturing processing methods.

[0069] Dietary supplements are products that are taken orally and contain the compounds or extracts of the present disclosure, and are intended to supplement the diet. Dietary supplements are products derived from food sources that provide additional health benefits in addition to the basic nutritional value contained in the food. Pharmaceutical compositions are defined as any component of a pharmaceutical product intended to provide pharmacological activity or other direct effects in the diagnosis, cure, mitigation, treatment, or prevention of disease, or to affect the structure or any function of the human or other animal body. Dietary supplements, nutraceuticals, and pharmaceutical compositions can be found in many forms, such as capsules, tablets, coated tablets, pills, capsules, pellets, granules, softgels, gelcaps, liquids, powders, emulsions, suspensions, elixirs, syrups, and any other form suitable for use.

[0070] The pharmaceutical compositions disclosed herein can be manufactured in a manner known per se, for example, by conventional mixing, dissolving, granulating, dragee-making, wet-grinding, emulsifying, encapsulating, encapsulating or tabletting processes.In addition, the active ingredient is contained in an amount effective to achieve its intended purpose.Many of the compounds used in the pharmaceutical combinations disclosed herein can be provided as salts with pharma-ceutical compatible counterions.

[0071] There are multiple techniques in the art for administering compounds, salts, and / or compositions, including, but not limited to, oral, rectal, pulmonary, topical, aerosol, injection, infusion, and parenteral delivery, including intramuscular, subcutaneous, intravenous, intramedullary injection, intrathecal, direct intraventricular, intraperitoneal, intranasal, and intraocular injection. In some embodiments, the compounds described herein, including those of formula (I), (II), (III), or pharma- ceutical acceptable salts thereof, can be administered orally.

[0072] Also, the compound, salt, and / or composition can be administered in a local rather than systemic manner, for example, by injecting or implanting the compound directly into the affected area, often in a depot or sustained release formulation.Furthermore, the compound can be administered in a targeted drug delivery system, for example, in liposomes coated with tissue-specific antibodies.The liposomes will target and be taken up selectively by organs.For example, intranasal or pulmonary delivery may be desired to target respiratory diseases or conditions.

[0073] The composition may be presented in a pack or dispenser device, which may contain one or more unit dosage forms containing the active ingredient, if desired. The pack may comprise, for example, metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also bear a notice associated with the container in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, which reflects the approval by the agency of the form of the drug for human or veterinary administration. Such notice may be, for example, a label approved by the U.S. Food and Drug Administration for prescription drugs, or an approved product insert. Compositions may also be prepared, placed in a suitable container, and labeled for the treatment of an indicated condition, which may comprise the compounds and / or salts described herein formulated in a compatible pharmaceutical excipient.

[0074] The compounds, salts, and / or pharmaceutical compositions may be provided to the administering physician or other medical practitioner in the form of a kit. The kit is a package that includes a container containing the compound(s) in a suitable pharmaceutical composition and instructions for administering the pharmaceutical composition to a subject. The kit may also optionally include one or more additional therapeutic agents. The kit may also include separate doses of the compound(s) or pharmaceutical composition for sequential or sequential administration. The kit may optionally include one or more diagnostic tools and instructions for use. The kit may include a suitable delivery device, such as a syringe, along with instructions for administering the compound(s) and any other therapeutic agents. The kit may optionally include instructions for storage, reconstitution (if applicable), and administration of any or all of the included therapeutic agents. The kit may include multiple containers reflecting the number of doses to be given to the subject.

[0075] In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is administered at a dose ranging from about 0.1 to 200 mg / kg body weight. In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is present in an amount of about 0.1 to 1, 0.5 to 1, 0.1 to 10, 0.5 to 10, 1 to 10, 1 to 20, 1 to 30, 1 to 40, 1 to 50, 1 to 60, 1 to 70, 1 to 80, 1 to 90, 1 to 100, 1 to 200, 1 to 300, 1 to 400, 1 to 500, 1 to 600, 1 to 700, 1 to 800, 1 to 900, 1 to 1000, 1 to 11, 1 to 12, 1 to 13, 1 to 13, 1 to 14, 1 to 15, 1 to 16, 1 to 17, 1 to 18, 1 to 19, 10 to 20, 10 to 30, 1 0~40, 10~50, 10~60, 10~70, 10~80, 10~90, 10~100, 10~200, 10~-300, 10~400, 10~500, 10~600, 10~700, 10~800, 10~900, 10~1000, 20~30, 20~40, 20~50, 20~60, 20~70, 20~80, 20~90, 20~100, 20~200, 20~300, 20~400, 20~500, 20~600, 20~700, 20~800, 20~900, 20~1000, 30~40, 30~50, 3 0~60, 30~70, 30~80, 30~90, 30~100, 30~200, 30~300, 30~400, 30~500, 30~600, 30~700, 30~800, 30~900, 30~1000, 40~50, 40~60, 40~70, 40~80, 40~90, 40~100, 40~200, 40~300, 40~400, 40~500, 40~600, 40~700, 40~800, 40~900, 40~1000, 50~60, 50~70, 50~80, 50~90, 50~100, 50~200, 50~300, 50~400, 50~500, 50~600, 50~700, 50~800, 50~900, 60~70, 60~80, 60~90, 60~100, 60~200, 60~300, 60~400, 60~500, 60~600, 60~700, 60~800, 60~900, 60~1000, 70~80, 70~90, 70~100, 70~200, 70~300, 70~400, 70~500, 70~600, 70~700, 70~800, 70~900, 70~1000, 80~90, 80~100,The compound is administered at a dose in the range of 80-200, 80-300, 80-400, 80-500, 80-600, 80-700, 80-800, 80-900, 80-100, 90-100, 90-200, 90-300, 90-400, 90-500, 90-600, 90-700, 90-800, 90-900, 90-1000, 100-150, 100-200, 100-300, 100-400, 100-500, 100-600, 100-700, 100-800, 100-900, or 100-1000 mg / kg body weight. In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is administered at a concentration of about 0.01, 0.02, 0.03, 0.05, 0.07, 0.1, 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, , 18.5, 19, 19.5, 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24, 24.5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29, 29.5, 30, 30.5, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 80, 90, or 95 mg / kg body weight. In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is administered at a concentration of about 0.01, 0.02, 0.03, 0.05, 0.07, 0.1, 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 4, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24, 24 .5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29, 29.5, 30, 30.5, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40mg / m, 2In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is administered in a dose of about 0.01, 0.02, 0.03, 0.05, 0.07, 0.1, 0.25, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 8 , 19.5, 20, 20.5, 21, 21.5, 22, 22.5, 23, 23.5, 24, 24.5, 25, 25.5, 26, 26.5, 27, 27.5, 28, 28.5, 29, 29.5, 30, 30.5, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 mg / kg of subject body weight.

[0076] In some embodiments, the dose of a compound of Formula (I), Formula (II), or Formula (III) is about 0.1 mg to 10 mg, 0.1 mg to 25 mg, 0.1 mg to 30 mg, 0.1 mg to 50 mg, 0.1 mg to 75 mg, 0.1 mg to 100 mg, 0.5 mg to 10 mg, 0.5 mg to 25 mg, 0.5 mg to 30 mg, 0.5 mg to 50 mg, 0.5 mg to 75 mg, 0.5 mg to 100 mg, 1 mg to 10 mg, 1mg~25mg, 1mg~30mg, 1mg~50mg, 1mg~75mg, 1mg~100mg, 2mg~10mg, 2mg~25mg, 2mg~30mg, 2mg~50mg, 2mg~7 5mg, 2mg~100mg, 3mg~10mg, 3mg~25mg, 3mg~30mg, 3mg~50mg, 3mg~75mg, 3mg~100mg, 4mg~100mg, 5mg~10mg, 5mg ~25mg, 5mg~30mg, 5mg~50mg, 5mg~75mg, 5mg~300mg, 5mg~200mg, 7.5mg~15mg, 7.5mg~25mg, 7.5mg~30mg, 7.5m g~50mg, 7.5mg~75mg, 7.5mg~100mg, 7.5mg~200mg, 10mg~20mg, 10mg~25mg, 10mg~50mg, 10mg~75mg, 10mg~100m g, 15 mg to 30 mg, 15 mg to 50 mg, 15 mg to 100 mg, 20 mg to 20 mg, 20 mg to 100 mg, 30 mg to 100 mg, 40 mg to 100 mg, 10 mg to 80 mg, 15 mg to 80 mg, 20 mg to 80 mg, 30 mg to 80 mg, 40 mg to 80 mg, 10 mg to 60 mg, 15 mg to 60 mg, 20 mg to 60 mg, 30 mg to 60 mg, or about 40 mg to 60 mg. In some embodiments, the compound of formula (I), formula (II), or formula (III) administered is about 20 mg to 60 mg, 27 mg to 60 mg, 20 mg to 45 mg, or 27 mg to 45 mg. In some embodiments, the compound of formula (I), formula (II), or formula (III) administered is about 10 mg to about 120 mg. In some embodiments, the compound of Formula (I), Formula (II), or Formula (III) is administered in an amount of about 1 mg to 5 mg, 1 mg to 7.5 mg, 2.5 mg to 5 mg, 2.5 mg to 7.5 mg, 5 mg to 7.5 mg, 5 mg to 9 mg, 5 mg to 10 mg, 5 mg to 12 mg, 5 mg to 14 mg,5mg~15mg、5mg~16mg、5mg~18mg、5mg~20mg、5mg~22mg、5mg~24mg、5mg~26mg、5mg~28mg、5mg~30mg、5mg~32mg、5mg~34mg、5mg~36mg、5mg~38mg、5mg~40mg、5mg~42mg、5mg~44mg、5mg~46mg、5mg~48mg、5mg~50mg、5mg~52mg、5mg~54mg、5mg~56mg、5mg~58mg、5mg~60mg、7mg~7.7mg、7mg~9mg、7mg~10mg、7mg~12mg、7mg~14mg、7mg~15mg、7mg~16mg、7mg~18mg、7mg~20mg、7mg~22mg、7mg~24mg、7mg~26mg、7mg~28mg、7mg~30mg、7mg~32mg、7mg~34mg、7mg~36mg、7mg~38mg、7mg~40mg、7mg~42mg、7mg~44mg、7mg~46mg、7mg~48mg、7mg~50mg、7mg~52mg、7mg~54mg、7mg~56mg、7mg~58mg、7mg~60mg、9mg~10mg、9mg~12mg、9mg~14mg、9mg~15mg、9mg~16mg、9mg~18mg、9mg~20mg、9mg~22mg、9mg~24mg、9mg~26mg、9mg~28mg、9mg~30mg、9mg~32mg、9mg~34mg、9mg~36mg、9mg~38mg、9mg~40mg、9mg~42mg、9mg~44mg、9mg~46mg、9mg~48mg、9mg~50mg、9mg~52mg、9mg~54mg、9mg~56mg、9mg~58mg、9mg~60mg、10mg~12mg、10mg~14mg、10mg~15mg、10mg~16mg、10mg~18mg、10mg~20mg、10mg~22mg、10mg~24mg、10mg~26mg、10mg~28mg、10mg~30mg、10mg~32mg、10mg~34mg、10mg~36mg、10mg~38mg、10mg~40mg、10mg~42mg、10mg~44mg、10mg~46mg、10mg~48mg、10mg~50mg、10mg~52mg、10mg~54mg、10mg~56mg、10mg~58mg、10mg~60mg、12mg~14mg、12mg~15mg、12mg~16mg、12mg~18mg、12mg~20mg、12mg~22mg、12mg~24mg、12mg~26mg、12mg~28mg、12mg~30mg、12mg~32mg、12mg~34mg、12mg~36mg、12mg~38mg、12mg~40mg、12mg~42mg、12mg~44mg、12mg~46mg、12mg~48mg、12mg~50mg、12mg~52mg、12mg~54mg、12mg~56mg、12mg~58mg、12mg~60mg、15mg~16mg、15mg~18mg、15mg~20mg、15mg~22mg、15mg~24mg、15mg~26mg、15mg~28mg、15mg~30mg、15mg~32mg、15mg~34mg、15mg~36mg、15mg~38mg、15mg~40mg、15mg~42mg、15mg~44mg、15mg~46mg、15mg~48mg、15mg~50mg、15mg~52mg、15mg~54mg、15mg~56mg、15mg~58mg、15mg~60mg、17mg~18mg、17mg~20mg、17mg~22mg、17mg~24mg、17mg~26mg、17mg~28mg、17mg~30mg、17mg~32mg、17mg~34mg、17mg~36mg、17mg~38mg、17mg~40mg、17mg~42mg、17mg~44mg、17mg~46mg、17mg~48mg、17mg~50mg、17mg~52mg、17mg~54mg、17mg~56mg、17mg~58mg、17mg~60mg、20mg~22mg、20mg~24mg、20mg~26mg、20mg~28mg、20mg~30mg、20mg~32mg、20mg~34mg、20mg~36mg、20mg~38mg、20mg~40mg、20mg~42mg、20mg~44mg、20mg~46mg、20mg~48mg、20mg~50mg、20mg~52mg、20mg~54mg、20mg~56mg、20mg~58mg、20mg~60mg、22mg~24mg、22mg~26mg、22mg~28mg、22mg~30mg、22mg~32mg、22mg~34mg、22mg~36mg、22mg~38mg、22mg~40mg、22mg~42mg、22mg~44mg、22mg~46mg、22mg~48mg、22mg~50mg、22mg~52mg、22mg~54mg、22mg~56mg、22mg~58mg、22mg~60mg、25mg~26mg、25mg~28mg、25mg~30mg、25mg~32mg、25mg~34mg、25mg~36mg、25mg~38mg、25mg~40mg、25mg~42mg、25mg~44mg、25mg~46mg、25mg~48mg、25mg~50mg、25mg~52mg、25mg~54mg、25mg~56mg、25mg~58mg、25mg~60mg、27mg~28mg、27mg~30mg、27mg~32mg、27mg~34mg、27mg~36mg、27mg~38mg、27mg~40mg、27mg~42mg、27mg~44mg、27mg~46mg、27mg~48mg、27mg~50mg、27mg~52mg、27mg~54mg、27mg~56mg、27mg~58mg、27mg~60mg、30mg~32mg、30mg~34mg、30mg~36mg、30mg~38mg、30mg~40mg、30mg~42mg、30mg~44mg、30mg~46mg、30mg~48mg、30mg~50mg、30mg~52mg、30mg~54mg、30mg~56mg、30mg~58mg、30mg~60mg、33mg~34mg、33mg~36mg、33mg~38mg、33mg~40mg、33mg~42mg、33mg~44mg、33mg~46mg、33mg~48mg、33mg~50mg、33mg~52mg、33mg~54mg、33mg~56mg、33mg~58mg、33mg~60mg、36mg~38mg、36mg~40mg、36mg~42mg、36mg~44mg、36mg~46mg、36mg~48mg、36mg~50mg、36mg~52mg、36mg~54mg、36mg~56mg、36mg~58mg、36mg~60mg、40mg~42mg、40mg~44mg、40mg~46mg、40mg~48mg、40mg~50mg、40mg~52mg、40mg~54mg、40mg~56mg、40mg~58mg、40mg~60mg、43mg~46mg、43mg~48mg、43mg~50mg、43mg~52mg、43mg~54mg、43mg~56mg、43mg~58mg、42mg to 60mg, 45mg to 48mg, 45mg to 50mg, 45mg to 52mg, 45mg to 54mg, 45mg to 56mg, 45mg to 58mg, 45mg to 60mg, 48mg to 50mg, 48mg to 52mg, 48mg to 54mg, 48mg to 56mg, 48mg to 58mg, 48mg to 60mg, 50mg to 52mg, 50mg to 54mg, 50mg to 56mg, 50mg to 58mg, 50mg to 60mg, 52mg to 54mg, 52mg to 56mg, 52mg to 58mg, or 52mg to 60mg. In some embodiments, the dose of the compound of Formula (I), Formula (II), or Formula (III) is 0.1 mg, 0.3 mg, 0.5 mg, 0.75 mg, 1 mg, 1.25 mg, 1.5 mg, 1.75 mg, 2 mg, 2.5 mg, 3 mg, 3.5 mg, 4 mg, 5 mg, about 10 mg, about 12.5 mg, about 13.5 mg, about 15 mg, about 17.5 mg, about 20 mg, about 22.5 mg, about Greater than, equal to, or about 25 mg, about 27 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, or about 1000 mg. In some embodiments, the dose of the compound of Formula (I), Formula (II), or Formula (III) is less than about 0.5 mg, 0.75 mg, 1 mg, 1.25 mg, 1.5 mg, 1.75 mg, 2 mg, 2.5 mg, 3 mg, 3.5 mg, 4 mg, 5 mg, about 10 mg, about 12.5 mg, about 13.5 mg, about 15 mg, about 17.5 mg, about 20 mg, about 22.5 mg, about 25 mg, about 27 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, or about 200 mg.

[0077] As used herein, the term "carrier" refers to a material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., lubricants, magnesium talc, calcium or zinc stearate, or stearic acid), or solvent encapsulating material, that is involved in carrying or transporting the subject compounds from one organ or part of the body to another. Each carrier must be compatible with the other ingredients of the formulation and not deleterious to the subject. Some examples of materials which can function as carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, cellulose acetate, and hydroxylpropylmethylcellulose; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; and (10) cellulose acetate. glycols, such as propylene glycol, (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol, (12) esters, such as ethyl oleate and ethyl laurate, (13) agar, (14) buffers, such as magnesium hydroxide and aluminum hydroxide, (15) alginic acid, (16) pyrogen-free water, (17) isotonic saline, (18) Ringer's solution, (19) ethyl alcohol, (20) pH buffer solutions, (21) polyesters, polycarbonates, and / or polyanhydrides, and (22) other non-toxic compatible substances used in conventional formulations.

[0078] To prepare solid compositions such as tablets or capsules, the compound or extract is mixed with carriers (e.g., conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, or gums) and other diluents (e.g., water) to form a solid composition. This solid composition is then subdivided into unit dosage forms containing an effective amount of the compound of the present disclosure. Tablets or pills containing the compound or extract can be coated or otherwise compounded to provide a dosage form offering the advantage of prolonged action.

[0079] In certain embodiments of the present disclosure, the consumer product composition comprises a compound or extract for reducing or retarding microbial growth, a carrier, and a preservative. In some embodiments, the preservative is added in an amount of up to about 5%. In some embodiments, the composition comprises about 0.01%, 0.1%, 1%, 2%, 3%, 4%, 5% preservative, or ranges including and / or spanning the aforementioned values. In some embodiments, the preservative is about 0.01% to about 1% by weight of the composition. In some embodiments, the preservative is about 1% to about 5% by weight of the composition. Examples of preservatives include, but are not limited to, sodium benzoate, methylparaben, propylparaben, sodium nitrite, sulfur dioxide, sodium sorbate, and potassium sorbate. Other suitable preservatives include, but are not limited to, salts of edetate (also known as salts of ethylenediaminetetraacetic acid, or EDTA, e.g., disodium EDTA).

[0080] The liquid forms that the compounds or extracts of the present disclosure are incorporated into for oral or parenteral administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions that contain edible oils, as well as elixirs and similar vehicles.Suitable dispersants or suspending agents for aqueous suspensions include synthetic natural gums, such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone, or gelatin.The liquid preparations for oral administration can take the form of, for example, solutions, syrups, or suspensions, or they can be provided as dry products for reconstitution with water or other suitable vehicles before use. Such liquid preparations can be prepared by conventional means with acceptable additives such as suspending agents (e.g., sorbitol syrup, methylcellulose, or hydrogenated edible fats), emulsifying agents (e.g., lecithin or acacia), non-aqueous vehicles (e.g., almond oil, oily esters, or ethyl alcohol), preservatives (e.g., methyl or propyl p-hydroxybenzoates or sorbic acid), and artificial or natural colors and / or sweeteners.

[0081] The method of preparing the formulation or composition of the present disclosure includes the step of bringing the compound or extract of the present disclosure into association with the carrier and, optionally, one or more accessory ingredients and / or active ingredients.In general, the formulation is prepared by uniformly and intimately bringing into association the compound or extract of the present disclosure with a liquid carrier or a finely divided solid carrier, or both, and then, if necessary, shaping the product.Thus, the disclosed formulation can consist of, or consist essentially of, the compound or extract described herein in combination with a suitable carrier.

[0082] When the compounds or extracts of the present disclosure are administered to humans and animals as pharmaceuticals, nutraceuticals, or dietary supplements, they may be given by themselves or as compositions containing, for example, 0.1-99% of the active ingredient in combination with an acceptable carrier. In some embodiments, the compounds or extracts of the present disclosure may be administered at about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% w / w, or ranges including and / or spanning the aforementioned values.

[0083] The consumable article may be ingested by a subject and provides less than 100 mg of a compound disclosed herein per day. In certain embodiments, the consumable article provides 10-60 mg / day of tyramine containing hydroxycinnamic acid amide. In certain embodiments, the consumable article provides 10-120 mg / day of tyramine containing hydroxycinnamic acid amide. In certain embodiments, the consumable article provides 100-200 mg / day of tyramine containing hydroxycinnamic acid amide. Effective amounts can be established by methods known in the art and may depend on bioavailability, toxicity, and the like.

[0084] While it is contemplated that individual tyramine containing hydroxycinnamic acid amides may be used in the consumable articles of the present disclosure, it is further contemplated that two or more of the compounds or extracts may be combined in any relative amounts to produce a conventional combination of ingredients containing two or more tyramine containing hydroxycinnamic acid amides in a desired ratio to enhance the efficacy of the product, improve the organoleptic properties or some other measure of quality important to the end use of the product.

[0085] combination Some aspects relate to combinations of a compound of formula (I), (II), or (III) with one or more compounds selected from dihydrosphingosine, ceramide, glycosphingolipids, and sphingosine. In some embodiments, the combination includes more compounds selected from dihydroceramide, ceramide, or sphingosine.

[0086] In some embodiments, the ceramide is selected from the group consisting of natural ceramides, synthetic ceramides, ceramide phosphates, 1-O-acyl-ceramides, dihydroceramides, dihydroceramide phosphates, and 2-hydroxyceramides.

[0087] In some embodiments, the natural ceramide is from pig brain or eggs.

[0088] In some embodiments, the synthetic ceramide is N-octadecanoyl-D-erythro-sphingosine (C18), N-hexadecanoyl-D-erythro-sphingosine (C16) N-acetoyl-D-erythro-sphingosine (C2 ceramide, d18:1 / 2:0), N-butyroyl-D-erythro-sphingosine (C4 ceramide, d18:1 / 4:0), N-hexanoyl-D-erythro-sphingosine (C6 ceramide, d18:1 / 6:0), N-octanoyl-D-erythro-sphingosine (C8 ceramide, d18:1 / 8:0), N- Decanoyl-D-erythro-sphingosine (C10 ceramide, d18:1 / 10:0), N-lauroyl-D-erythro-sphingosine (C12 ceramide, d18:1 / 12:0), N-myristoyl-D-erythro-sphingosine (C14 ceramide, d18:1 / 14:0), N-palmitoyl-D-erythro-sphingosine (C16 ceramide: d18:1 / 16:0), N-heptadecanoyl-D-erythro-sphingosine (C17 ceramide: d18:1 / 17:0), N-stearoyl-D-erythro-sphingosine (C18 ceramide, d18:1 / 18:0), N-oleoyl-D-erythro-sphingosine (C18:1 ceramide, d18:1 / 18:1(9Z)), N-arachidoyl-D-erythro-sphingosine (C20 ceramide, d18:1 / 20:0), N-behenoyl-D-erythro-sphingosine (C22 ceramide, d18:1 / 22:0), N-lignoceroyl-D-erythro-sphingosine (C24 ceramide, d18:1 / 24:0), N-nervonoyl-D-erythro-sphingosine (C24:1 ceramide, d18:1 / 24:1(15Z)), N-acetoacetate N-Octanoyl-D-erythro-sphingosine (C17 base) (C2 ceramide, d17:1 / 2:0), N-Octanoyl-D-erythro-sphingosine (C17 base) (C8 ceramide, d17:1 / 8:0), N-Stearoyl-D-erythro-sphingosine (C17 base) (C18 ceramide, d17:1 / 18:0), N-Oleoyl-D-erythro-sphingosine (C17 base) (C18:1 ceramide, d17:1 / 18:1(9Z)), N-Arachidoyl-D-erythro-sphingosine (C17 base) (C20 ceramide, d17:1 / 20:0),N-lignoceroyl-D-erythro-sphingosine (C17 base) (C24 ceramide, d17:1 / 24:0) and N-nervonoyl-D-erythro-sphingosine (C17 base) (C24:1 ceramide, d17:1 / 24:1(15Z)).

[0089] In some embodiments, the ceramide phosphate is N-acetoyl-ceramide-1-phosphate (ammonium salt) (C2 ceramide-1-phosphate, d18:1 / 2:0), N-octanoyl-ceramide-1-phosphate (ammonium salt) (C8 ceramide-1-phosphate, d18:1 / 8:0), N-lauroyl-ceramide-1-phosphate (ammonium salt) (C12 ceramide-1-phosphate, d18:1 / 12:0), N-palmitoyl-ceramide-1-phosphate (ammonium salt) (C16 ceramide-1-phosphate, d18:1 / 16 ...palmitoyl-ceramide-1-phosphate (ammonium salt) (C16 ceramide-1-phosphate, d18:1 / 16:0), N-palmitoyl-ceramide-1-phosphate (ammonium salt) (C16 ceramide-1-phosphate, d18:1 / 16:0), N-palmitoyl-ceramide-1-phosphate (ammonium salt) (C1 :0), N-oleoyl-ceramide-1-phosphate (ammonium salt) (C18:1 ceramide-1-phosphate, d18:1 / 18:1(9Z), N-lignoceroyl-ceramide-1-phosphate (ammonium salt) (C24 ceramide-1-phosphate, 18:1 / 24:0), N-acetoyl-ceramide-1-phosphate (C17 base) (C2 ceramide-1-phosphate, d17:1 / 2:0), and N-octanoyl-ceramide-1-phosphate (C17 base) (C8 ceramide-1-phosphate, d17:1 / 8:0).

[0090] In some embodiments, the dihydroceramide is N-hexanoyl-D-erythro-sphinganine (C6 dihydroceramide, d18:0 / 6:0), N-octanoyl-D-erythro-sphinganine (C8 dihydroceramide, d18:0 / 8:0), N-palmitoyl-D-erythro-sphinganine (C16 dihydroceramide, d18:0 / 16:0), N-stearoyl-D-erythro-sphinganine (C18 dihydroceramide, d18:0 / 18:0), N-oleoyl-D-erythro-sphinganine (C18:1 dihydroceramide, d18:0 / 18:1(9Z)), N-lignoceroyl-D-erythro-sphinganine (C24 dihydroceramide, d18:0 / 24:0), and N-nervonoyl-D-erythro-sphinganine-D-erythro-sphinganine (C24:1 dihydroceramide, d18:0 / 24:1(15Z)).

[0091] In some embodiments, the dihydroceramide phosphate is N-palmitoyl-D-erythro-dihydroceramide-1-phosphate (ammonium salt) (C16 dihydroceramide-1-phosphate, d18:0 / 16:0) or N-lignoceroyl-D-erythro-dihydroceramide-1-phosphate (ammonium salt) (C24 dihydroceramide-1-phosphate, d18:0 / 24:0).

[0092] In some embodiments, the 2-hydroxyceramide is N-(2'-(R)-hydroxylauroyl)-D-erythro-sphingosine (12:0(2R-OH)ceramide), N-(2'-(S)-hydroxylauroyl)-D-erythro-sphingosine (12:0(2S-OH)ceramide), N-(2'-(R)-hydroxypalmitoyl)-D-erythro-sphingosine (16:0(2R-OH)ceramide), N-(2'-(S)-hydroxypalmitoyl)-D-erythro-sphingosine (16:0(2S-OH)ceramide). ceramide), N-(2'-(R)-hydroxyheptadecanoyl)-D-erythro-sphingosine (17:0(2R-OH)ceramide), N-(2'-(S)-hydroxyheptadecanoyl)-D-erythro-sphingosine (17:0(2S-OH)ceramide), N-(2'-(R)-hydroxystearoyl)-D-erythro-sphingosine (18:0(2R-OH)ceramide), N-(2'-(S)-hydroxystearoyl)-D-erythro-sphingosine (18:0(2S-OH)ceramide), N-(2'-(R)-hydroxystearoyl)-D-erythro-sphingosine (18:0(2S-OH)ceramide), N-(2'-(S)-hydroxyoleoyl)-D-erythro-sphingosine (18:1(2R-OH)ceramide), N-(2'-(S)-hydroxyoleoyl)-D-erythro-sphingosine (18:1(2S-OH)ceramide), N-(2'-(R)-hydroxyarachidoyl)-D-erythro-sphingosine (20:0(2R-OH)ceramide), N-(2'-(S)-hydroxyarachidoyl)-D-erythro-sphingosine (20:0(2S-OH)ceramide), N-(2'-(R)-hydroxybehenoyl)-D-erythro-sphingosine N-(2'-(S)-hydroxylbehenoyl)-D-erythro-sphingosine (22:0(2S-OH)ceramide), N-(2'-(R)-hydroxylignoceroyl)-D-erythro-sphingosine (24:0(2R-OH)ceramide), N-(2'-(S)-hydroxylignoceroyl)-D-erythro-sphingosine (24:0(2S-OH)ceramide), N-(2'-(R)-hydroxynervonoyl)-D-erythro-sphingosine (24:1(2R-OH)ceramide),and N-(2'-(S)-hydroxyl nervonoyl)-D-erythro-sphingosine (24:1 (2S-OH) ceramide).

[0093] In some embodiments, the sphingosine is selected from the group consisting of natural sphingosine, synthetic sphingosine, phosphorylated sphingosine (S1P), and methylated sphingosine.

[0094] In some embodiments, the naturally occurring sphingosine is D-erythro-sphingosine.

[0095] In some embodiments, the synthetic sphingosine is selected from the group consisting of sphingosine (d18:1), sphingosine (d17:1), sphingosine (d20:1), L-threo-sphingosine (d18:1), 1-deoxysphingosine, and 1-desoxymethylsphingosine. In some embodiments, the sphinganine is selected from the group consisting of sphinganine (d18:0), sphinganine (d17:0), sphinganine (d20:0), 1-deoxysphinganine, 1-desoxymethylsphinganine, and L-threo-dihydrosphingosine (d18:0) (safingol). In some embodiments, the phosphorylated sphingosine is selected from the group consisting of sphingosine-1-phosphate (d18:1), sphingosine-1-phosphate (DMA adduct), sphingosine-1-phosphate (d17:1), sphingosine-1-phosphate (d20:1), sphinganine-1-phosphate (d18:0), sphinganine-1-phosphate (d17:0), and sphinganine-1-phosphate (d20:0). In some embodiments, the methylated sphingosine is selected from the group consisting of monomethylsphingosine (d18:1), dimethylsphingosine (d18:1), dimethylsphingosine (d17:1), trimethylsphingosine (d18:1), trimethylsphingosine (d17:1), dimethylsphinganine (d18:0), trimethylsphinganine (d18:0), dimethylsphingosine-1-phosphate (d18:1), and dimethylsphinganine-1-phosphate (d18:0).

[0096] In some embodiments, the glycosphingolipid is selected from the group consisting of natural glycosphingolipids, glycosylsphingolipids, galactosylsphingolipids, lactosylsphingolipids, sulfatides, and α-galactosylceramide (αGalCer).

[0097] In some embodiments, the naturally occurring glycosphingolipid is selected from the group consisting of cerebroside (e.g., from pig brain), glucocerebroside (e.g., from soybean), sulfatide (ammonium salt) (e.g., from pig brain), GM1 ganglioside (ammonium salt) (e.g., from sheep brain), ganglioside GM1 (e.g., from sheep brain), and total ganglioside extract (ammonium salt) (e.g., from pig brain).

[0098] In some embodiments, the glycosylsphingolipid is D-glucosyl-β1-1′-D-erythro-sphingosine (glucosyl(β)sphingosine, d18:1), D-glucosyl-β-1,1′N-octanoyl-D-erythro-sphingosine (C8 glucosyl(β)ceramide, d18:1 / 8:0), D-glucosyl-β-1,1′N-lauroyl-D-erythro-sphingosine (C12 glucosyl(β)ceramide, d18:1 / 12:0), D-glucosyl-β-1,1′N-palmito ... D-glucosyl-β-1,1′N-stearoyl-D-erythro-sphingosine (C18 glucosyl(β)ceramide, d18:1 / 18:0), D-glucosyl-β-1,1′N-oleoyl-D-erythro-sphingosine (C18:1 glucosyl(β)ceramide, d18:1 / 18:1(9Z)), and D-glucosyl β1-1′-N-nervonoyl-D-erythro-sphingosine (C24:1 glucosyl(β)ceramide, dl 8:1 / 24:1(15Z)).

[0099] In some embodiments, the galactosyl sphingolipid is D-galactosyl-β1-1′-D-erythro-sphingosine (galactosyl(β)sphingosine, d18:1), N,N-dimethyl-D-galactosyl-β1-1′-D-erythro-sphingosine (galactosyl(β)dimethylsphingosine, d18:1), D-galactosyl-β-1,1′N-octanoyl-D-erythro-sphingosine (C8 galactosyl(β)ceramide, d18:1 / 8:0), D-galactosyl-β1-1′ ... Tosyl-β-1,1′N-lauroyl-D-erythro-sphingosine (C12 galactosyl(β)ceramide, d18:1 / 12:0), D-galactosyl-β-1,1′N-palmitoyl-D-erythro-sphingosine (C16 galactosyl(β)ceramide, d18:1 / 16:0), and D-galactosyl-β-1,1′-N-norbornoyl-D-erythro-sphingosine (C24:1 galactosyl(β)ceramide, d18:1 / 24:1(15Z)).

[0100] In some embodiments, the lactosyl sphingolipid is D-lactosyl-β1-1′-D-erythro-sphingosine (lactosyl(β)sphingosine, d18:1), D-lactosyl-β-1,1′N-octanoyl-D-erythro-sphingosine (C8 lactosyl(β)ceramide, d18:1 / 8:0), D-lactosyl-β1-1′-N-octanoyl-D-threo-sphingosine (C8 L-threo-lactosyl(β)ceramide, d18:1 / 8:0), D-lactosyl-β-1,1′N-lauroyl-D-erythro-sphingosine (C12 lactosyl(β)ceramide, d18:1 / 12:0), D-lactosyl-β-1,1′N-palmitoyl-D-erythro-sphingosine (C16 lactosyl(β)ceramide, d18:1 / 16:0), D-lactosyl-β-1,1′N-lignoceroyl-D-erythro-sphingosine (C24 lactosyl(β)ceramide, d18:1 / 24:0), and D-lactosyl-β1-1′-N-nervonoyl-D-erythro-sphingosine (C24:1 lactosyl(β)ceramide, d18:1 / 24:1).

[0101] In some embodiments, the sulfatide is 3-O-sulfo-D-galactosyl-β1-1′-N-lignoceroyl-D-erythro-sphingosine (ammonium salt) (e.g., from pig brain), 3-O-sulfo-D-galactosyl-β1-1′-N-lauroyl-D-erythro-sphingosine (ammonium salt) (C12 mono-sulfogalactosyl(β)ceramide, d18:1 / 12:0), 3-O-sulfo-D-galactosyl-β1-1′-N-heptadecanoyl-D-erythro-sphingosine (ammonium salt) (C17 mono-sulfogalactosyl(β)ceramide, d18:1 / 17:0), 3-O-sulfo-D-galactosyl-β1-1′-N-heptadecanoyl-D-erythro-sphingosine (ammonium salt) (C17 mono-sulfogalactosyl(β)ceramide, d18:1 / 17:0), 3-O-sulfo-D- Galactosyl-β1-1′-N-lignoceroyl-D-erythro-sphingosine (ammonium salt) (C24 mono-sulfogalactosyl (β) ceramide (d18:1 / 24:0), 3-O-sulfo-D-galactosyl-β1-1′-N-nervonoyl-D-erythro-sphingosine (ammonium salt) (C24:1 mono-sulfogalactosyl (β) ceramide, d18:1 / 24:1), and 3,6-di-O-sulfo-D-galactosyl-β1-1′-N-lauroyl-D-erythro-sphingosine (ammonium salt) (C12 di-sulfogalactosyl (β) ceramide, d18:1 / 12:0).

[0102] In some embodiments, the phosphosphingolipid is selected from the group consisting of D-erythro-sphingosylphosphoethanolamine (sphingosyl PE, d18:1), N-lauroyl-D-erythro-sphingosylphosphoethanolamine (C17 base) (C12 sphingosyl PE, d17:1 / 12:0), and D-erythro-sphingosylphosphoinositol (sphingosyl PI).

[0103] In some embodiments, phytosphingosine is selected from the group consisting of 4-hydroxysphinganine (Saccharomyces cerevisiae) (D-ribo-phytosphingosine), 4-hydroxysphinganine (C17 base) (D-ribo-phytosphingosine, C17 base), 4-hydroxysphinganine-N,N-dimethyl (Saccharomyces cerevisiae) (phytosphingosine-N,N-dimethyl), 4-hydroxysphinganine-N,N,N-trimethyl (methyl sulfate salt) (Saccharomyces cerevisiae) (phytosphingosine-N,N,N-trimethyl), 4-hydroxysphinganine-1-phosphate (Saccharomyces cerevisiae) (D-ribo-phytosphingosine-1-phosphate), 4-hydroxysphinganine-N,N-dimethyl-1-phosphate (ammonium salt) (Saccharomyces cerevisiae) (phytosphingosine-N,N-dimethyl-1-phosphate), N-acetoyl 4-hydroxysphinganine (saccharomyces cerevisiae) (N-02:0 phytosphingosine), N-octanoyl 4-hydroxysphinganine (saccharomyces cerevisiae) (N-08:0 phytosphingosine), N-stearoyl 4-hydroxysphinganine (saccharomyces cerevisiae) (N-18:0 phytosphingosine), and 4-hydroxysphinganine-1-phosphocholine (saccharomyces cerevisiae) (phytosphingosine phosphocholine).

[0104] Some aspects relate to combinations of a compound of formula (I), (II), or (III) with one or more compounds selected from macrolides, retinides, and DES1 inhibitors. In some embodiments, the one or more retinides are fenretinide, N-(4-hydroxyphenyl)retinamide (4-HPR), 4-oxo-N-(4-hydroxyphenyl)retinamide (4-oxo-HPR), or motretinide. In some embodiments, the DES1 inhibitor is selected from N-[(1R,2S)-2-hydroxy-1-hydroxymethyl-2-(2-tridecyl-1-cyclopropenyl)ethyl]octanamide (GT011) and (Z)-4-((5-(4-chlorophenyl)-1,3,4-oxadiazol-2-yl)amino)-N'-hydroxybenzimidamide (B-0027). In some embodiments, the one or more macrolides are selected from the group consisting of rapamycin, erythromycin, clarithromycin, roxithromycin, azithromycin, fidaxomicin, carbomycin A, josamycin, kitasamycin, midecamycin, oleandomycin, solithromycin, spiramycin, troleandomycin, tyrosine, roxithromycin, telithromycin, cethromycin, solithromycin, solithromycin, tacrolimus, pimecrolimus, sirolimus, cyclosporine, polyene antifungals, and cluentaren.

[0105] How to use The present disclosure provides for promoting an increase in mitochondrial mass by providing a consumer product composition as described herein and at least one carrier. Such a method provides an effective amount of a composition as described herein to a subject in need thereof, thereby enhancing mitochondrial mass and function in the subject. As used herein, the term "subject" refers to an animal, preferably a mammal. In some embodiments, the subject is a veterinary, companion, farm, laboratory, or zoological animal. In other embodiments, the subject is a human.

[0106] In some aspects, administering a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt, isomer, homodimer, heterodimer, or conjugate thereof, increases mitochondrial mass. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a disease or condition associated with enhanced mitochondrial mass in a subject. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a disease or condition associated with enhanced mitochondrial mass in a subject. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a disease or condition associated with enhanced mitochondrial mass and function in a subject.

[0107] In one embodiment, administering a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, treats or improves at least one factor associated with mitochondrial mass and function in a subject. In other aspects, a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, disclosed herein, enhances mitochondrial mass and function in a subject, e.g., by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%, or a range including and / or spanning the foregoing values. In yet another aspect, a composition comprising Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, can be used to improve mitochondrial mass and function, for example, about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about 10% to about 10 ... The improvement or enhancement is in the range of 0%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.

[0108] In some embodiments, an effective amount of a composition described herein is provided to a subject in need thereof, thereby increasing fatty acid oxidation in the subject. As used herein, the term "subject" refers to an animal, preferably a mammal. In some embodiments, the subject is a veterinary, companion, farm, laboratory, or zoological animal. In other embodiments, the subject is a human.

[0109] In some aspects, administering a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt, isomer, homodimer, heterodimer, or conjugate thereof, increases fatty acid oxidation in a subject. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a disease or condition associated with increased fatty acid oxidation in a subject. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a disease or condition associated with increased fatty acid oxidation in a subject. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a disease or condition associated with increased fatty acid oxidation in a subject.

[0110] In one embodiment, administering a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, treats or ameliorates at least one factor associated with increased fatty acid oxidation in a subject. In other aspects, a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, disclosed herein, increases fatty acid oxidation in a subject by, for example, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%, or a range including and / or spanning the foregoing values. In yet another embodiment, a composition comprising Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, enhances fatty acid oxidation at, for example, about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 90%, about The increase is in the range of 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.

[0111] In one embodiment, administering a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, treats or ameliorates at least one factor associated with a high calorie diet in a subject. In other aspects, a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, disclosed herein, treats fatty acid excess in a subject, for example, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%, or ranges including and / or spanning the foregoing values. In yet another embodiment, a composition comprising Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, can be used to reduce one or more signs of aging in a subject in need thereof, for example, about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 9 ...100%, about 10% to about 100%, about 10% to about 100%, about 10% to about 100%, about 10% to about 100%, about 10% to about 100%, about % to about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.In still another aspect, a composition comprising Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, provides cellular energy to a subject in need thereof, for example, about 10% to about 100%, about 20% to about 100%, about 30% to about 100%, about 40% to about 100%, about 50% to about 100%, about 60% to about 100%, about 70% to about 100%, about 80% to about 100%, about 10% to about 90%, about 20% to about 100%, or about 15% to about 15%. about 90%, about 30% to about 90%, about 40% to about 90%, about 50% to about 90%, about 60% to about 90%, about 70% to about 90%, about 10% to about 80%, about 20% to about 80%, about 30% to about 80%, about 40% to about 80%, about 50% to about 80%, or about 60% to about 80%, about 10% to about 70%, about 20% to about 70%, about 30% to about 70%, about 40% to about 70%, or about 50% to about 70%.

[0112] In some embodiments, an effective amount of the composition described herein is provided to a subject in need thereof, thereby increasing NAD in the subject. As used herein, the term "subject" refers to an animal, preferably a mammal. In some embodiments, the subject is a veterinary, companion, farm, laboratory, or zoological animal. In other embodiments, the subject is a human.

[0113] In some aspects, administering a composition comprising a compound of formula (I), formula (II), or formula (III), or a pharma- ceutically acceptable salt, isomer, homodimer, heterodimer, or conjugate thereof, increases NAD in a subject. In some embodiments, a composition comprising a compound of formula (I), formula (II), or formula (III) treats or improves a disease or condition associated with increased NAD in a subject. In some embodiments, a composition comprising a compound of formula (I), formula (II), or formula (III) treats or improves a disease or condition associated with increased NAD in a subject. In some embodiments, a composition comprising a compound of formula (I), formula (II), or formula (III) treats or improves a disease or condition associated with increased NAD in a subject.

[0114] In one embodiment, administering a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, treats or ameliorates at least one factor associated with increased NAD in a subject. In other aspects, a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, disclosed herein, increases NAD in a subject by, for example, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%, or a range including and / or spanning the foregoing values.

[0115] In some embodiments, an effective amount of the composition described herein is provided to a subject in need thereof, thereby inhibiting ceramide to dihydroceramide conversion in the subject. As used herein, the term "subject" refers to an animal, preferably a mammal. In some embodiments, the subject is a veterinary, companion, farm, laboratory, or zoological animal. In other embodiments, the subject is a human.

[0116] In some aspects, administering a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt, isomer, homodimer, heterodimer, or conjugate thereof inhibits ceramide to dihydroceramide conversion in a subject. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a ceramide-associated disease or condition in a subject. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a ceramide-associated disease or condition in a subject. In some embodiments, a composition comprising a compound of Formula (I), Formula (II), or Formula (III) treats or improves a ceramide-associated disease or condition in a subject.

[0117] In one embodiment, administering a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, treats or ameliorates at least one factor associated with inhibition of ceramide to dihydroceramide conversion in a subject. In other aspects, a composition comprising a compound of Formula (I), Formula (II), or Formula (III), or a pharma- ceutically acceptable salt thereof, disclosed herein, inhibits ceramide to dihydroceramide conversion in a subject by, for example, at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%, or a range including and / or spanning the aforementioned values.

[0118] In some embodiments, the subject is on a high fat diet. In some embodiments, the subject is on a high calorie diet. In some embodiments, the subject is obese. In some embodiments, the subject is obese prior to receiving an oral composition described herein. In some embodiments, the subject is overweight. In some embodiments, the subject is overweight prior to receiving an oral composition described herein.

[0119] In some embodiments, the subject is administered the oral composition for at least 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 3 months, 6 months, 1 year, or ranges including and / or spanning the foregoing values. In some embodiments, the subject is administered the oral composition for at least 6 weeks. In some embodiments, the subject is administered the oral composition for at least 10 weeks.

[0120] In some embodiments, the subject's body weight is reduced by about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, or a range including and / or spanning the foregoing values, hi some embodiments, the subject's body weight is reduced by about 2% to about 20%.

[0121] In some aspects, the disclosure provides a method of treating, preventing, improving, or reducing muscle wasting or muscle wasting disorders in a subject suffering from such disorders by administering an effective amount of a composition described herein. Muscle wasting disorders or conditions include, but are not limited to, muscular dystrophies (such as DMD, Becker MD, limb-girdle MD, myotonic MD, and FSHD), myositis (such as dermatomyositis, inclusion body myositis, juvenile myositis, polymyositis, and the like), myopathy (including inherited and acquired myopathies, such as diabetic myopathy or drug-induced myopathy), motor neuron disease (such as Lou Gehrig's disease or amyotrophic lateral sclerosis), myasthenia gravis, neurodegenerative disorders (such as Parkinson's disease, Huntington's disease, and Alzheimer's disease), muscle wasting associated with cancer (such as pancreatic cancer, lung cancer, gastric cancer, ovarian cancer, colorectal cancer, melanoma, and the like). leukemia, lung cancer, prostate cancer, brain cancer, bladder cancer, and head and neck cancer), chronic heart failure (CHF), chronic kidney disease (CKD), liver failure, diabetes, chronic obstructive pulmonary disease (COPD), emphysema, cystic fibrosis, rheumatoid arthritis, osteoarthritis, liver fibrosis, muscle wasting associated with cirrhosis, trauma (such as burns or motorcycle accidents), fractures, organ transplants (such as heart, lung, liver, or kidney transplants), ICU critical care, denervation (such as stalk or spinal cord injury), androgen deprivation therapy, corticosteroid therapy, infections (such as AIDS and tuberculosis), prolonged bed rest, sarcopenic obesity, and age-related sarcopenia. In some embodiments, muscle wasting is reduced in the subject by about, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%, or a range including and / or spanning the foregoing values. In some embodiments, the subject is on a high fat diet.

[0122] In some aspects, the disclosure provides methods of increasing or improving performance recovery in a subject in need thereof by administering an effective amount of a composition described herein. In some embodiments, performance recovery can include increased mobility in the subject. In some embodiments, performance recovery can include increased endurance in the subject. In some embodiments, performance recovery can include increased strength or power in the subject. In some embodiments, performance recovery can include increased speed in the subject. In some embodiments, performance is improved or increased in the subject by about, e.g., at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%, or ranges including and / or spanning the foregoing values. In some embodiments, the subject is on a high fat diet. EXAMPLES

[0123] The following non-limiting examples are provided to further illustrate the present disclosure.

[0124] Example 1 HNF4α is a nuclear receptor transcription factor that controls the expression of downstream genes that are important in multiple aspects of cellular metabolism. The classical view of HNF4α is that its ligand binding pocket (LBP) is constitutively occupied by fatty acids that play a structural but not regulatory role. However, it has recently been shown that fatty acids in the HNF4α LBP are exchangeable, particularly in cells, in the context of full-length HNF4α. Using a cell-based promoter reporter assay for human insulin promoter activity that is highly sensitive to HNF4α activity, it has been demonstrated that fatty acids act as HNF4α antagonists. Using the assay in a high-throughput screen, HNF4α antagonists and more recently agonists have been discovered. Recently, a potent agonist, N-trans-caffeoyltyramine, has been discovered. The availability of potent HNF4α agonists has facilitated in vivo studies that have revealed novel aspects of HNF4α biology. Previously, intraperitoneal (IP) administration of NCT was shown to reverse hepatic steatosis in obese mice fed a high-fat diet through a previously unsuspected pathway involving HNF4α regulation of dihydroceramide synthesis. Dihydroceramide was found to regulate hepatic lipophagy, leading to the reversal of hepatic steatosis.

[0125] Here, another previously unsuspected effect of HNF4α was revealed, made possible by the discovery that HNF4α can be delivered orally mixed with a high-fat diet, allowing for long-term NCT administration. As in previous studies of IP NCT administration, a reduction in hepatic steatosis was observed. However, mice on a high-fat diet administered NCT for 10 weeks were much lower in body weight than those on a high-fat diet alone. The reduction in fat appeared to be due to an increase in fatty acid oxidation, which in turn was attributed to an increase in mitochondrial mass. Consistent with that, there was an increase in the expression of mitochondrial proteins, including VDAC1 and electron transport chain proteins, mitochondrial DNA, and total cellular NAD, most of which resides in the mitochondria. There was a marked reduction in markers of inflammation and cellular stress, including nitric oxide. Thus, NCT is a strong candidate for a drug that can maintain metabolic homeostasis in the face of problems from excessive fatty acid intake.

[0126] In vivo mouse experiments Four-week-old male C57BL / 6J (JAX catalog number 000664) mice were purchased from Jackson laboratory and maintained on a 12-hour light / day cycle throughout the experiment. Mice were allowed to acclimate for 2 weeks prior to the experiment. Six-week-old mice with similar body weights were randomized to a standard diet (NC), a high-fat diet (HFD) (Research Diets, catalog number D12492 60 kcal% fat), or a HFD containing 4000 ppm of NCT (HFD+NCT) (Research Diets, 60 kcal% fat + 4000 ppm of NCT), calculated to provide approximately 400 mg / kg / day of NCT. The HFD diet containing NCT was made with a gray dye to distinguish it from the regular HFD diet, which has a green dye. Mice in each treatment group were placed in separate cages. All mice were provided with equal amounts of fresh chow every week. Weight gain and food intake were measured weekly for 10 weeks. After 10 weeks of dietary treatment, mice were sacrificed for analysis. All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of the Sanford Burnham Prebys Medical Discovery Institute in accordance with national regulations. Diet size was selected based on prior studies of NCT delivered by IP injection.

[0127] Subcutaneous and oral gavage treatments Twelve-week-old C57BL / 6J DIO male mice were purchased from Jackson laboratory (cat. no. 380050) and fed a high-fat diet (Research Diets, cat. no. D12492 60 kcal% fat). Mice were allowed to acclimate for 2 weeks prior to the experiment. Mice at 14 weeks of age were subcutaneously injected twice daily for 2 weeks using a sterile insulin syringe loaded with DMSO or NCT (200 mg / kg mouse body weight). For oral gavage, 14-week-old C57BL / 6J DIO male mice were fed by oral gavage twice daily for 2 weeks using a feeding needle attached to a sterile 1 ml syringe loaded with 200 μL methylcellulose (MC, vehicle control) or NCT (200 mg / kg) dissolved in 200 μL methylcellulose. All mice were maintained on a 12-hour light / day cycle throughout the experiment. For analysis, all mice were sacrificed after 2 weeks of treatment.

[0128] Sample collection Mouse samples were collected as previously described. Briefly, on the last day of treatment, mice received dextrose (3 g / kg body weight) by IP injection to stimulate insulin secretion, which inhibits the release of FFA from adipocytes, leaving liver-derived FFA as the major source of circulating FFA. After 1 h, blood samples were collected via retro-orbital bleeding, and mice were euthanized using pentobarbital. Mice were aseptically dissected, and liver, epididymal fat, and body weight were measured and photographed. Dissected liver samples were immediately washed in sterile cold PBS and cut into small pieces. Half of the liver samples were snap frozen using liquid nitrogen and stored at -80°C for RNA, protein isolation, and liver lysate preparation. The remaining half was fixed in 4% cold paraformaldehyde (PFA, Santa Cruz Biotechnology, USA) and processed for histomorphometry and immunofluorescence.

[0129] Oil Red O staining (in vivo) and analysis: Oil Red O staining was performed as previously described (34). Slides containing frozen liver tissue sections from mice were air-dried for 10–20 min and then rehydrated in distilled water. Sections were immersed in anhydrous propylene glycol (catalog no. 151957, MP Biomedicals, LLC, USA) for 2 min, followed by immersion in 0.5% in Oil Red O solution (catalog no. K043, Poly Scientific R&D, USA) for 2 h. Slides were then differentiated in 85% propylene glycol solution and diluted with dHO. 2 The slides were washed with Oil Red O for 2 h and fixed using glycerin jelly mounting medium. All slides were scanned at 20x magnification using an Aperio Scanscope FL system (Aperio Technologies Inc., Vista, CA, USA). The liver area stained with Oil Red O was measured using Image J software as described, and the Oil Red O stained liver images were opened in Image J software with some modifications as follows: The scale bar of the images was set to 200 um using the Analyze > Set Scale command. The RGB images were then converted to grayscale images using the Image > Type > RGB Stack command and split into red, blue, and green channels. The threshold was manually set to highlight the Oil Red O stained lipid droplets in the green channel using the Image > Adjust > Threshold command. The same threshold was used for all images of all treatment groups, and the Oil Red O stained area % was obtained using the Analyze → Measure Tools command. The fold change was calculated by normalizing the values ​​to images from mice fed a standard diet.

[0130] Triglyceride Analysis TG levels in the liver, serum, and feces of mice were measured using a Triglyceride Calorimetry Assay Kit (Cat. No. 10010303, Cayman Chemicals, USA) according to the manufacturer's instructions. Liver and fecal TG were normalized to the weight of liver and feces, respectively. Fold changes were calculated by normalizing to the values ​​from mice fed NC.

[0131] Free fatty acid quantification Blood samples were collected from dextrose-injected mice and centrifuged to collect serum samples. Serum FFA levels were measured using a Free Fatty Acid Quantification Colorimetric / Fluorometric Kit (Cat. No. K612, BioVision, USA) according to the manufacturer's instructions. Fold changes were calculated by normalizing to values ​​from NC-fed mice.

[0132] Immunofluorescence Frozen liver sections were permeabilized using 0.3% Triton-X and incubated in antigen retrieval solution (antigen retrieval citrate, Biogenex) for 10 min at sub-boiling temperature. Sections were then incubated with blocking buffer (Jackson Immuno Research) containing 5% normal donkey serum, followed by incubation with mouse anti-HNF4α monoclonal antibody (1:800, Catalog No. PP-H1415-00, R&D Systems), rabbit polyclonal anti-VDAC antibody (1:400, Catalog No. PA1-954A, Invitrogen), and cleaved caspase 3 antibody (1:500, Catalog No. 9664, Cell Signaling) at 4°C overnight. Sections were washed and incubated with anti-mouse secondary antibody conjugated with Alexa fluor 488 (1:400, Invitrogen) and anti-rabbit secondary antibody conjugated with rhodamine red for 1 h at room temperature. Nuclei were visualized by counterstaining with DAPI (40,6-diamidino-2-phenylindole, Sigma Aldrich). Slides were fixed using fluorescent mounting medium and images were acquired at 40x magnification using an Olympus IX71 fluorescent microscope. Fluorescence intensities of HNF4α-stained nuclei and VDAC-stained mitochondria were calculated using MetaMorph TL software (version 7.6.5.0, Olympus). Fold changes were calculated by normalizing to values ​​from NC-fed mice.

[0133] Liver profile analysis Mice were anesthetized and 100 μL of whole blood was collected via retro-orbital bleed into lithium heparin blood collection tubes and transferred to a single-use VetScan mammalian liver profile reagent rotor. Levels of multiple analytes, including alkaline phosphatase (ALP), alanine aminotransferase (ALT), gamma glutamyl transferase (GGT), bile acids (BA), TBIL (total bilirubin), albumin (ALB), blood urea nitrogen 371 (BUN), and total cholesterol (CHOL), were quantified using a VetScan VS2 Chemistry Analyzer (Abaxis North America, USA).

[0134] blood cell count analysis Mice were anesthetized, 20 μL of whole blood was collected via retro-orbital bleeding into lithium heparin blood collection tubes, and 20 different hematological parameters were measured using a Hemavet 950 FS blood cell count analyzer (Drew Scientific Group).

[0135] Western blotting Mouse liver extracts were prepared by incubation in RIPA buffer (Invitrogen) containing protease inhibitors (Calbiochem, San Diego, CA). Proteins were quantified by BCA assay (Thermo Scientific). Proteins (40 mg or 80 mg for cytochrome C) were separated on 12% or 16% triglycine gels (Invitrogen) and transferred to Immobilon P membranes (0.2 μm pore size, Millipore). After 1 h in phosphate buffered saline-Tween (PBST) with 3% milk, the membranes were incubated with antibodies against PPARGC1A (PGC1α) (cat. no. NBP1-04676, Novus, 1:1000), SDHA XP (cat. no. 11998, Cell Signaling 1:1000), or cytochrome C (cat. no. 4280, Cell Signaling 1:500), followed by incubation with secondary antibodies conjugated to horseradish peroxidase (1:5000, Jackson Immune or Cell Signaling). Signals were revealed by ECL (Thermo) and imaged with a ChemiDoc MP imager (Bio-Rad). After detection, the membranes were incubated with Ponceau S solution (Sigma) for 1 h to normalize to loaded protein.

[0136] Primary Human Hepatocytes Experiments with primary human hepatocytes were performed by CN-Bio (Cambridge, UK). Briefly, primary human hepatocytes (PHH), human Kupffer cells (HK), and human stellate cells (HSC) were cultured at 6x10 for PHH in 1.6 ml of CN-Bio's HEP lean media containing 5% FCS. 5 cells, 6x10 for HK and HSC 4Cells were seeded in PhysioMimix LC12 MPS culture plates from CN-Bio at 100 μl / s. Cells were maintained at a flow rate of 1 μl / sec throughout the experiment. 24 hours after seeding (day 1), the medium was changed to HEP low fat medium and cells were incubated until day 4 to allow the cells to form microtissues. Four days after seeding, the medium was changed to HEP fat medium and treated with DMSO or NCT (5, 15, 40 μM). Medium was changed on days 6 and 8. Cells were harvested on day 10 for RNA extraction and culture medium was collected for ELISA analysis.

[0137] RT-PCR Total RNA was isolated from liver tissues and primary human hepatocytes using Trizol (Invitrogen). cDNA was amplified using 3 μg of total RNA using qScript cDNA SuperMix (Quanta BioSciences, Beverly, MA, USA). Quantitative real-time PCR (RT-PCR) analysis was performed using SYBR® Select Master Mix (Applied Biosystems) and an ABI 7900HT thermal cycler (Applied Biosystems, Thermo Fisher Scientific). Ct values ​​were normalized to 18s rRNA and expressed as fold change relative to samples from mice fed NC or from human hepatocytes cultured without NCT.

[0138] Fatty acid oxidation assay FAO in liver lysates was measured using a calorimetric assay kit (catalog number E-141, Biomedical Research Service Center, State University of New York, Buffalo, NY) according to the manufacturer's instructions.

[0139] Nicotinamide adenine dinucleotide assay Total NAD levels in liver lysates were analyzed using a calorimetric NAD+ / NADH assay kit (catalog no. MET-5014, Cell Biolabs, Inc. USA) according to the manufacturer's instructions.

[0140] Citrate synthase activity Citrate synthase activity in liver homogenates was measured using the calorimetric-based MitoCheck Citrate Synthase Activity Assay Kit (cat. no. 701040, Cayman Chemicals, USA) according to the manufacturer's instructions.

[0141] BCA assay Bicinchoninic acid (BCA) protein assay was performed using a kit from Thermo Scientific (cat. no. 23225) according to the manufacturer's instructions. The BCA assay was used for protein quantification in the FAO assay and Western blotting. The absorbance at 550 nm was determined using a plate reader.

[0142] Nitric oxide assay T6PNE cells were incubated at 37 °C for 24 h at 5% CO 2 Cells were maintained at 37°C in RPMI (5.5 mM glucose, Corning) supplemented with 10% fetal bovine serum (FBS, Sigma-Aldrich) and 1% penicillin-streptomycin (pen-strep, Gibco). Cells were treated with 0.12 μM palmitate + 0 or 15 μM NCT for 3 days in 10 cm plates and harvested in 500 μl PBS. For tissue preparations, flash-frozen mouse livers were weighed and homogenized in PBS. NO was measured with a QuantiChrom Nitric Oxide Assay kit (D2NO-100, BioAssay Systems). Homogenized samples (150 μl) were diluted with 8 μl ZnSO 4and 8 μl NaOH for deproteinization. For normalization, an aliquot of sample from T6PNE cells was taken for BCA assay before deproteinization. Samples and standards from the kit were incubated with the reagent for 20 min at 60°C and OD540nM was measured.

[0143] Palmitate-BSA Complex Palmitate (150 mM) (Sigma-Aldrich) was prepared in 50% ethanol and precomplexed with 15% fatty acid-free BSA (Research Organics, Cleveland, OH, USA) in a water shaker at 37° C. BSA447-complexed palmitate was used as a 12 mM stock solution for all assays with a final concentration of 0.12 mM palmitate in cell culture medium.

[0144] Mitochondrial DNA analysis Quantification of mtDNA was performed as described (BCurr Protoc Mouse Biol 2017;7:47-54). Snap-frozen mouse livers were homogenized and total cellular DNA was extracted using the QIAamp DNA kit (Qiagen), followed by qPCR using the following primers: 16S rRNA primer FWD: 5′-CCGCAAGGGAAAGATGAAAGAC-3′ REV:5′-TCGTTTGGTTTCGGGGTTTC-3′ ND1 primer FWD: 5′-CTAGCAGAAACAAACCGGGC-3′ REV:5′-CCGGCTGCGTATTCTACGTT-3′ HK2 primer FWD: 5′-GCCAGCCTCTCCTGATTTTAGTGT-3′ REV:5′-GGGAACACAAAAGACCTCTTCTGG-3

[0145] statistical analysis Data are presented as the mean ± SEM of triplicate samples, as indicated. Statistical significance was assessed using Student's t test or ANOVA.

[0146] Long-term administration of NCT led to weight loss Previously, we performed a short 2-week study in which NCT, a potent HNF4α agonist, was administered to obese mice. NCT induced a reduction in hepatic steatosis by stimulation of lipophagy, but no effect on body weight was observed. In that study, NCT was administered intraperitoneally (IP) due to the hydrophobic nature of HNF4α ligands, including natural fatty acid ligands and the non-natural ligands against which it was tested. This resulted in compound precipitation when attempts were made to deliver NCT subcutaneously (SQ) (Figure 6A-6D - NCT subcutaneous and oral gavage delivery resulted in compound precipitation: 14-week-old male C57BL / 6 DIO mice received two doses of 200 mg / kg NCT per day for 2 weeks via subcutaneous injection) or by oral gavage (Figure 6E-I - 14-week-old male C57BL / 6 DIO mice received two doses of methylcellulose (MC, vehicle control) or NCT (200 mg / kg) per day via oral gavage) for 2 weeks. Following oral gavage, NCT was present in the feces but not in the serum (Figure 6I), indicating malabsorption of the compound that left the stomach (Figure 6F). Given the desirability of oral administration, delivery of NCT mixed with a high-fat diet mouse chow (HFD) (Research Diets) was tested. A HFD containing 4,000 ppm NCT (HFD+NCT) (Research Diets) was calculated to provide lean mice weighing approximately 20 gm with approximately the same dose of NCT (400 mg / kg / day) as delivered using the previously used IP administration protocol. HFD+NCT was administered to 6-week-old C57BL / 6 male mice for 10 weeks. Figures 1A-1N illustrate data from an experiment of chronic administration of N-trans-caffeoyltyramine that resulted in reduced high-fat diet-induced weight gain and hepatic steatosis (C57BL / 6 mice fed standard chow (NC), HFD, or HFD+NCT during the 10-week study). To determine whether orally administered NCT was active, we examined its effect on hepatic HNF4α expression, which was previously found to be increased by NCT. HFD led to a decrease in HNF4α expression, which was reversed by NCT, consistent with previous results when NCT was administered IP (Figures 1A and 1B).Thus, oral administration of NCT at doses similar to those administered IP was effective in stimulating HNF4α activity.

[0147] In sharp contrast to previous experiments with 2 weeks of IP NCT administration, where no effect of NCT on body weight was observed, it became apparent at week 5 that mice fed HFD+NCT weighed less than mice fed HFD alone (Figure 1C). By the end of the 10-week experiment, mice fed HFD+NCT weighed approximately 10 gm less than mice fed HFD, a 35-40% difference in body weight. One possible explanation for the lower body weight was that the mice consumed less food, possibly due to the long-term toxicity of NCT. There were no discernible differences in physical activity or other behaviors between HFD-fed and HFD+NCT-fed mice (Figure 10). Caspase 3, a marker of apoptosis, was lower in HFD-fed mice and unchanged with NCT (Figure 7A-7D). T6PNE cells were treated with or without palmitate (0.1 mM) and 0, 5, 10, and 20 µM NCT for 3 days, followed by fixation in 4% PFA and immunostaining for cleaved caspase 3 and DAPI. Quantification was performed using a Celigo imaging cytometer (Nexcelom Bioscience). There was no difference in the amount of HFD and HFD+NCT feed ingested, indicating that NCT was not aversive and did not cause mice to become sick and ingest less food (Figure 1D). Fecal triglycerides (TG) were comparable in the HFD and HFD+NCT groups, ruling out the possibility of fat malabsorption being the cause of the differences in body weight (Figure 1E). HFD+NCT mice were significantly less obese (Fig. 1F) and had substantially less subcutaneous fat (Fig. 1N), but there was no change in epididymal fat pad weight at the end of the study (Fig. 1L and Fig. 1M). HFD+NCT mice had redder livers (Fig. 1G) and lower liver weights (Fig. 1H). Consistent with increased redness and reduced body weight, HFD+NCT livers showed reduced Oil Red O staining (Fig. 1I, quantified in Fig. 1J) and lower TG (Fig. 1K). There were no differences in circulating TG or free fatty acids (FFA) (Fig. 8A-8C - NCT did not modulate TG, FFA, and ALP levels in the circulation).

[0148] NCT led to increased fatty acid oxidation (FAO) in the liver of obese mice with hepatic steatosis, but this was a secondary, not a primary, effect The large difference in body weight between the HFD and HFD+NCT groups was notable along with a reduction in subcutaneous and hepatic adiposity. There are only two routes of fat elimination: fecal via malabsorption (which was excluded) (Figure 1E) and fecal via oxidation. It was therefore hypothesized that NCT induces an increase in FAO. Quantification of FAO was performed using an assay that measures NADH by the conversion of iodonitrotetrazolium (INT) to INT-formazan, mediated by the enzyme diaphorase, which requires NADH. Performing the assay in the presence and absence of added octanoyl-CoA, which is converted to acetyl-CoA via fatty acid β-oxidation with conversion of NAD+ to NADH, provides specificity to FAO. Fatty acid oxidation levels are determined as the INT-formazan levels in the presence of octanoyl-CoA minus the levels in the absence of octanoyl-CoA.

[0149] NCT induced an increase in FAO activity in the presence of octanoyl-CoA (Figure 2A). However, there was also an increase in baseline activity in the absence of octanoyl-CoA (Figure 2B), resulting in no change in overall activity in the assay (Figure 2C). Because the assay is sensitive to cellular mitochondrial mass, this suggests that the effect of NCT may not be specific to fatty acid oxidation but on mitochondrial mass, i.e., NCT may act to stimulate an increase in mitochondrial mass with a consequent increase in total fatty acid oxidation without stimulating an increase in fatty acid oxidation levels per unit of mitochondrial mass. Independent measurements of total cellular NAD, which is mostly located in mitochondria, revealed a substantial increase in the liver of mice treated with NCT (Figure 2D). Thus, the effect of NCT on FAO appeared to be a secondary rather than a primary effect.

[0150] NCT induced an increase in mitochondrial mass and reduced mitochondrial stress Based on the results of oxidation of FAO and the increase in total cellular NAD, tested markers of mitochondrial mass directly determined whether NCT led to an increase. The two most commonly used protein markers of mitochondrial mass are VDAC1, located in the outer mitochondrial membrane (11), and citrate synthase, located in the mitochondrial matrix (12). HFD led to a significant decrease in VDAC1 expression and citrate synthase activity, and these decreases were substantially reversed by NCT ​​(Figure 3A-C).

[0151] The major function of mitochondria is oxidative phosphorylation. To determine whether the increased mitochondrial mass was reflected in increased expression of proteins involved in oxidative phosphorylation, cytochrome C and succinate dehydrogenase expression were measured, both of which are important components of the respiratory apparatus. Cytochrome C plays a dual role in mitochondria, being important for mitochondrial respiration but also playing a role in cell survival. Succinate dehydrogenase, encoded by the SDHA gene, is the catalytic subunit of succinate-ubiquinone oxidoreductase, a complex of the mitochondrial respiratory chain. HFD reduced hepatic cytochrome C and succinate dehydrogenase protein levels, which was reversed by NCT ​​(Figures 3D-F). NCT increased SDHA, but not cytochrome C mRNA, in mouse liver (Figures 3G and 3H). This was reflected in studies with primary human hepatocytes, where NCT increased levels of SDHA but not cytochrome C mRNA (Figures 3I and 3J).

[0152] The increase in mitochondrial protein could be due to an increase in the number of mitochondria or simply due to an increase in the size of mitochondria. Therefore, we measured mitochondrial DNA content and found that NCT induced a significant increase in mitochondrial DNA, as measured by the levels of DNA encoding the mitochondrial genes ND1 and 16S rRNA (Figure 3K and Figure 3L).

[0153] A key feature of NAFLD and its progression to NASH is mitochondrial stress. To determine whether the reduction in hepatic steatosis and increase in mitochondrial mass induced by NCT, translated to 167, reduced mitochondrial stress, we examined the expression of HSP60, a mitochondrial chaperone induced by mitochondrial stress including HFD (Figure 3M). The level of HSP60 mRNA was significantly reduced by NCT, demonstrating the alleviation of mitochondrial stress (Figure 3M). Similarly, PPARγ plays a key role in the mitochondrial stress response, activated by fatty acids and showing high expression in fatty liver disease. NCT administration dramatically reduced PPARγ expression to the level in the liver of mice fed a standard chow diet (Figure 3N).

[0154] NCT increased the activity of the PPARGC1A pathway The ability of NCT to increase mitochondrial mass raised the question of the mechanism by which it occurred. Examination of RNA-seq data from the livers of mice treated with NCT for 2 weeks revealed a ~6-fold increase in PPARGC1A (PGC1a) mRNA (GEO 174848). PPARGC1A plays an important role in mitochondrial biogenesis but is not known to be regulated by HNF4α. After 10 weeks of oral NCT administration, PPARGC1A protein and mRNA were increased in mouse liver (Figure 4A-C). PPARGC1A mRNA levels in primary human hepatocytes cultured with NCT were also elevated (Figure 4F).

[0155] PPARGC1A activity is controlled by sirtuins, which are NAD-dependent deacetylases. Both of these are downstream targets of transcriptional activation by PPARGC1A and activators of PPARGC1A activity by deacetylation. HFD reduced Sirt1 and Sirt3 mRNA levels, consistent with previous studies. NCT almost completely reversed the effect of HFD on sirtuin gene expression in mouse liver (Figure 4D and Figure 4E) and had a profound effect in primary human hepatocytes (Figure 4G and Figure 4H). Poly(ADP-ribose) polymerase (PARP) inhibits mitochondrial function and PPARGC1A activity. In contrast to sirtuin expression, Parp1 and Parp2 mRNA levels were not modulated by NCT ​​(Figure 9A-9B - no difference in Parp1 and Parp2 mRNA expression).

[0156] NCT inhibited inflammation Inflammation is a major factor in the pathophysiology of NAFLD and its progression to NASH. Inflammation contributes to essential features of NASH pathology, including fibrosis and hepatocyte death, ultimately leading to cirrhosis. IL-6 and TNFα are important inflammatory mediators in NASH. Both were significantly reduced by NCT ​​in livers of HFD+NCT mice compared to HFD mice livers (Figure 5A and Figure 5B). Similarly, IL-6 mRNA and protein were reduced by NCT ​​in cultured human hepatocytes (Figure 5D and Figure 5E). TNFα mRNA was also dose-responsively reduced by NCT ​​in cultured human hepatocytes (Figure 5F), whereas IL1β expression was not significantly altered (Figure 5C). Nitric oxide (NO) plays an important role in inflammatory responses, including NAFLD. NO was increased in livers of HFD-fed mice (Figure 5G), as well as in cultured cells treated with palmitate (Figure 5H), and was reduced by NCT ​​in both settings (Figure 5G and Figure 5H). Blood ALT levels were elevated in HFD-fed mice and significantly decreased by NCT ​​(Figure 5I). Blood ALP was unchanged (Figure 8C). Other components of the liver profile panel and hematological analysis showed no differences in HFD vs. HFD+NCT mice (Figures 11 and 12).

[0157] The main finding of the study presented here is that activating HNF4α with the potent HNF4α agonist NCT led to weight loss in mice fed a high-fat diet. This is due to a considerable increase in mitochondrial mass and a resulting increase in fatty acid oxidation. A previous study in which NCT was administered briefly to mice found a different effect, namely stimulation of lipophagy. This led to a reduction in hepatic steatosis, but no change in body weight, as fatty acids released from lipid vesicles by lipophagy seem to redistribute to adipose tissue. Thus, there appear to be at least two independent effects of HNF4α agonism on fat storage, namely an immediate effect on hepatic steatosis by lipophagy and a long-term effect on mitochondrial mass leading to an increase in mitochondrial function, including fatty acid oxidation.

[0158] Key to uncovering the effect of pharmacological activation of HNF4α on mitochondrial mass was the discovery that oral delivery of NCT in a high-fat diet was effective and allowed for long-term delivery of the compound. This was an important goal, as being able to deliver drugs orally is often crucial to achieving effective clinical application. Oral delivery is particularly important for highly prevalent chronic diseases like NAFLD / NASH. Previous studies of NCT used IP delivery because the compound is hydrophobic, limiting some routes of delivery. However, efficacy was demonstrated in the HFD-fed mouse model of NAFLD by combining NCT with a high-fat diet. Of note, there is a significant difference in the microsomal stability of NCT between rodents and humans, with NCT being much more stable in the presence of human microsomes than in rodents. This bodes well for eventual use in humans, but pharmacokinetic studies in the context of human clinical trials are needed. A potentially important consideration is that NCT and fatty acids are likely competing for the HNF4α ligand-binding pocket. The concentrations of fatty acids that act as HNF4α antagonists in the nucleus are unknown, but if related to the overall level of steatosis in a particular organ, this may influence the dose required for efficacy.

[0159] The results led to a 234 increase in mitochondrial mass with weight loss in mice fed HFD+NCT and in mice fed HFD alone. Since food intake was the same and there was no fat malabsorption, an increase in fatty acid oxidation alone can reasonably explain the lower body weight of HFD+NCT mice. This was evidenced by an increase in the oxidation of octanoyl-CoA in mice fed HFD+NCT. However, the increase in fatty acid oxidation was not selective, since there was an increase in NADH production even in the absence of octanoyl-CoA. Consistent with the general increase in mitochondrial mass and function, VDAC1 and citrate synthase were increased, as were the oxidative phosphorylation components cytochrome C and SDHA. Consistent with the general increase in mitochondrial function, total cellular NAD was significantly increased by NCT.

[0160] The pathways that control mitochondrial mass are complex and interlocked, and a protein at the center of those pathways is the transcriptional coactivator PPARGC1A (PGC1α). PPARGC1A interacts with multiple factors involved in mitochondrial biogenesis and function. Notably, it directly interacts with HNF4α in regulating gene expression, particularly gluconeogenesis (26), although HNF4α has not previously been recognized as having an effect on mitochondrial biogenesis. PPARGC1A expression was increased by NCT, but is not thought to be a direct HNF4α target based on ChIP-seq data. In addition, NCT increased the expression of sirtuins that control the level of PPARGC1A activity by deacetylation. Pharmacological enhancement of mitochondrial mass and function has long been a sought-after goal, but robust active compounds with the desired activity have been lacking. A major motivation for finding approaches to increase mitochondrial mass and function is that mitochondrial function declines with age. Given the discovery of increased mitochondrial mass and function with HNFα activation, NCT may be beneficial in that setting. However, a consideration with stimulating HNF4α activity is that it is expressed at high levels only in a subset of tissues, including the liver, intestine, pancreas, and kidney. Thus, the effects of HNF4α agonists may be limited to those organs, although there is evidence of low levels of HNF4α expression elsewhere. Furthermore, the effects of NCT extended to reducing obesity, despite the fact that HNF4α is not expressed in adipocytes. Furthermore, there are important age-related diseases that affect organs that express HNF4α. Most prominent among them is type 2 diabetes, where age and lipotoxic effects on pancreatic β cells over time are significant. Thus, NCT has potential as a type 2 diabetes treatment. The liver plays a key role in type 2 diabetes, and the main paradigm is that cellular stress and inflammation contribute to insulin resistance and dysregulated hepatic gluconeogenesis, and are also important in the progression of NAFLD to NASH. NCT was effective in reducing cellular stress and inflammation, supporting a role in multiple diseases in which these factors are central.

[0161] Important to the eventual clinical application of NCT, no adverse effects of this compound were observed in any of the short-term maximum tolerated dose studies, studies involving IP administration at the fairly high dose of 200 mg / kg twice daily for 2 weeks, and long-term oral administration. Given the chronic nature of lipotoxicity disorders, the lack of observed toxicity of NCT is important, but will clearly require further investigation in human clinical trials.

Claims

1. 1. A method for enhancing mitochondrial mass and function in a subject in need thereof, the method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of a compound of formula (I); 【Chemistry 1】 During the ceremony, R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , and R ９ are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted —(O)C １－６ Alkyl, optionally substituted -(O)C １－６ Alkenyl, optionally substituted -(O)C １－６ Alkynyl, optionally substituted -(O)C ４－１２ Cycloalkyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Cycloalkyl, optionally substituted -(O)C ４－１２ Heterocyclyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Heterocyclyl, optionally substituted -(O)C ４－１２ Aryl, optionally substituted -(O)C １－６ Alkyl C ５－１２ Aryl, optionally substituted -(O)C １－１２ Heteroaryl and optionally substituted -(O)C １－６ Alkyl C １－１２ heteroaryl, the dashed bond is present or absent; X is CH ２ or O, Z is CHR ａ , N.R. ａ or O; R ａ is hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C １－６ Alkyl, optionally substituted -(O)C １－６ Alkenyl, optionally substituted -(O)C １－６ Alkynyl, optionally substituted -(O)C ４－１２ Cycloalkyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Cycloalkyl, optionally substituted -(O)C ４－１２ Heterocyclyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Heterocyclyl, optionally substituted -(O)C ４－１２ Aryl, optionally substituted -(O)C １－６ Alkyl C ５－１２ Aryl, optionally substituted -(O)C １－１２ Heteroaryl and optionally substituted -(O)C １－６ Alkyl C １－１２ heteroaryl; The subject is on a high fat or high calorie diet.

2. 10. The method of claim 1, wherein the subject is administered the oral composition for at least six weeks.

3. 3. The method of claim 1 or 2, wherein the subject's body weight is reduced by about 2% to about 20%.

4. 3. The method of claim 1 or 2, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical composition.

5. 3. The method of claim 1 or 2, wherein the compound of formula (I) is selected from the group consisting of N-cis-caffeoyltyramine, N-trans-feruloyltyramine, N-cis-feruloyltyramine, p-coumaroyltyramine, cinnamoyltyramine, sinapoyltyramine, and 5-hydroxyferuloyltyramine.

6. 3. The method of claim 1 or 2, wherein the subject is administered about 10 mg to about 120 mg of the compound of the oral composition.

7. The method of claim 1 or 2, wherein the oral composition is formulated as a tablet or capsule.

8. 3. The method of claim 1 or 2, wherein the carrier is selected from at least one of sugars, starches, celluloses, powdered tragacanth, malt, gelatin, talc, excipients, oils, glycols, polyols, esters, agar, buffers, alginic acid, isotonic saline, ethyl alcohol, pH buffer solutions, or polyesters.

9. The method of claim 1 or 2, wherein the compound of formula (I) comprises from about 0.1% to about 99% of the composition.

10. 3. The method of claim 1 or 2, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical.

11. The method of claim 1 or 2, wherein the oral composition further comprises a preservative.

12. The method of claim 11, wherein the preservative is from about 0.01% to about 5% by weight of the composition.

13. The method of claim 1 or 2, wherein the oral composition has a pH of from 2 to 7.

4.

14. The method of claim 1 or 2, wherein the oral composition is formulated as a liquid.

15. 1. A method for increasing fatty acid oxidation in a subject in need thereof, the method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of a compound of formula (I); 【Chemistry 2】 During the ceremony, R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , and R ９ are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted —(O)C １－６ Alkyl, optionally substituted -(O)C １－６ Alkenyl, optionally substituted -(O)C １－６ Alkynyl, optionally substituted -(O)C ４－１２ Cycloalkyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Cycloalkyl, optionally substituted -(O)C ４－１２ Heterocyclyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Heterocyclyl, optionally substituted -(O)C ４－１２ Aryl, optionally substituted -(O)C １－６ Alkyl C ５－１２ Aryl, optionally substituted -(O)C １－１２ Heteroaryl and optionally substituted -(O)C １－６ Alkyl C １－１２ heteroaryl, the dashed bond is present or absent; X is CH ２ or O, Z is CHR ａ , N.R. ａ or O; R ａ is hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C １ - ６ Alkyl, optionally substituted -(O)C １－６ Alkenyl, optionally substituted -(O)C １ - ６ Alkynyl, optionally substituted -(O)C ４－１２ Cycloalkyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Cycloalkyl, optionally substituted -(O)C ４－１２ Heterocyclyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Heterocyclyl, optionally substituted -(O)C ４－１２ Aryl, optionally substituted -(O)C １－６ Alkyl C ５－１２ Aryl, optionally substituted -(O)C １－１２ Heteroaryl and optionally substituted -(O)C １－６ Alkyl C １－１２ heteroaryl; The subject is on a high fat or high calorie diet.

16. 16. The method of claim 15, wherein the subject is administered the oral composition for at least 6 weeks.

17. 17. The method of claim 15 or 16, wherein the subject's body weight is reduced by about 2% to about 20%.

18. 17. The method of claim 15 or 16, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical composition.

19. 17. The method of claim 15 or 16, wherein the compound of formula (I) is selected from the group consisting of N-cis-caffeoyltyramine, N-trans-feruloyltyramine, N-cis-feruloyltyramine, p-coumaroyltyramine, cinnamoyltyramine, sinapoyltyramine, and 5-hydroxyferuloyltyramine.

20. 17. The method of claim 15 or 16, wherein the subject is administered about 10 mg to about 120 mg of the compound of the oral composition.

21. 17. The method of claim 15 or 16, wherein the oral composition is formulated as a tablet or capsule.

22. 17. The method of claim 15 or 16, wherein the carrier is selected from at least one of sugars, starches, celluloses, powdered tragacanth, malt, gelatin, talc, excipients, oils, glycols, polyols, esters, agar, buffers, alginic acid, isotonic saline, ethyl alcohol, pH buffer solutions, or polyesters.

23. The method of claim 15 or 16, wherein the compound of formula (I) comprises from about 0.1% to about 99% of the composition.

24. 17. The method of claim 15 or 16, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical.

25. The method of claim 15 or 16, wherein the oral composition further comprises a preservative.

26. 26. The method of claim 25, wherein the preservative is from about 0.01% to about 5% by weight of the composition.

27. The method of claim 15 or 16, wherein the oral composition has a pH of 2 to 7.

4.

28. 17. The method of claim 15 or 16, wherein the oral composition is formulated as a liquid.

29. 1. A method for treating fatty acid excess in a subject in need thereof, the method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of N-trans-caffeoyltyramine; The subject is on a high fat or high calorie diet.

30. 30. The method of claim 29, wherein the subject is administered the oral composition for at least 6 weeks.

31. 31. The method of claim 29 or 30, wherein the subject's body weight is reduced by about 2 to about 20%.

32. 31. The method of claim 29 or 30, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical composition.

33. 1. A method for reducing one or more signs of aging in a subject in need thereof, the method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of N-trans-caffeoyltyramine; The subject is on a high fat or high calorie diet.

34. 34. The method of claim 33, wherein the subject is administered the oral composition for at least 6 weeks.

35. 35. The method of claim 33 or 34, wherein the subject's body weight is reduced by about 2 to about 20%.

36. 36. The method of any one of claims 33 to 35, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical composition.

37. 1. A method for increasing cellular energy in a subject in need thereof, said method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of N-trans-caffeoyltyramine; The subject is on a high fat or high calorie diet.

38. 38. The method of claim 37, wherein the subject is administered the oral composition for at least 6 weeks.

39. 39. The method of claim 37 or 38, wherein the subject's body weight is reduced by about 2 to about 20%.

40. 40. The method of any one of claims 37 to 39, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical composition.

41. 1. A method for providing an energy boost in a subject in need thereof, the method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of N-trans-caffeoyltyramine; The subject is on a high fat or high calorie diet.

42. 42. The method of claim 41, wherein the subject is administered the oral composition for at least six weeks.

43. 43. The method of claim 41 or 42, wherein the subject's body weight is reduced by about 2 to about 20%.

44. 44. The method of any one of claims 41 to 43, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical composition.

45. 1. A method for increasing NAD in a subject in need thereof, the method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of N-trans-caffeoyltyramine; The subject is on a high fat or high calorie diet.

46. 46. ​​The method of claim 45, wherein the subject is administered the oral composition for at least 6 weeks.

47. 47. The method of claim 45 or 46, wherein the subject's body weight is reduced by about 2 to about 20%.

48. 47. The method of any one of claims 45 to 46, wherein the oral composition is formulated as a dietary supplement, a food ingredient or additive, a medical food, a nutraceutical, or a pharmaceutical composition.

49. 1. A method for reducing muscle atrophy in a subject in need thereof, the method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of a compound of formula (I); 【Chemistry 3】 During the ceremony, R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , and R ９ are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted —(O)C １－６ Alkyl, optionally substituted -(O)C １－６ Alkenyl, optionally substituted -(O)C １－６ Alkynyl, optionally substituted -(O)C ４－１２ Cycloalkyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Cycloalkyl, optionally substituted -(O)C ４－１２ Heterocyclyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Heterocyclyl, optionally substituted -(O)C ４－１２ Aryl, optionally substituted -(O)C １－６ Alkyl C ５－１２ Aryl, optionally substituted -(O)C １－１２ Heteroaryl and optionally substituted -(O)C １－６ Alkyl C １－１２ heteroaryl, the dashed bond is present or absent; X is CH ２ or O, Z is CHR ａ , N.R. ａ or O; R ａ is hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C １－６ Alkyl, optionally substituted -(O)C １－６ Alkenyl, optionally substituted -(O)C １－６ Alkynyl, optionally substituted -(O)C ４－１２ Cycloalkyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Cycloalkyl, optionally substituted -(O)C ４－１２ Heterocyclyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Heterocyclyl, optionally substituted -(O)C ４－１２ Aryl, optionally substituted -(O)C １－６ Alkyl C ５－１２ Aryl, optionally substituted -(O)C １－１２ Heteroaryl and optionally substituted -(O)C １－６ Alkyl C １－１２ heteroaryl.

50. 1. A method for increasing performance recovery in a subject in need thereof, the method comprising: administering to said subject in need thereof an oral composition comprising at least one carrier and an effective amount of a compound of formula (I); 【Chemistry 4】 During the ceremony, R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , R ８ , and R ９ are each independently hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted —(O)C １－６ Alkyl, optionally substituted -(O)C １－６ Alkenyl, optionally substituted -(O)C １－６ Alkynyl, optionally substituted -(O)C ４－１２ Cycloalkyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Cycloalkyl, optionally substituted -(O)C ４－１２ Heterocyclyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Heterocyclyl, optionally substituted -(O)C ４－１２ Aryl, optionally substituted -(O)C １－６ Alkyl C ５－１２ Aryl, optionally substituted -(O)C １－１２ Heteroaryl and optionally substituted -(O)C １－６ Alkyl C １－１２ heteroaryl, the dashed bond is present or absent; X is CH ２ or O, Z is CHR ａ , N.R. ａ or O; R ａ is hydrogen, deuterium, hydroxyl, halogen, cyano, nitro, optionally substituted amino, optionally substituted C-amido, optionally substituted N-amido, optionally substituted ester, optionally substituted -(O)C １－６ Alkyl, optionally substituted -(O)C １－６ Alkenyl, optionally substituted -(O)C １－６ Alkynyl, optionally substituted -(O)C ４－１２ Cycloalkyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Cycloalkyl, optionally substituted -(O)C ４－１２ Heterocyclyl, optionally substituted -(O)C １－６ Alkyl C ４－１２ Heterocyclyl, optionally substituted -(O)C ４－１２ Aryl, optionally substituted -(O)C １－６ Alkyl C ５－１２ Aryl, optionally substituted -(O)C １－１２ Heteroaryl and optionally substituted -(O)C １－６ Alkyl C １－１２ heteroaryl.

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