RBP4 antagonists for treatment and prevention of non-alcoholic fatty liver disease and gout

Abstract

The present invention provides a method for treating non-alcoholic fatty liver disease (NAFLD) in a subject suffering from the disease, the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective for treating the subject, thereby treating the subject, the compound is a retinol binding protein 4 (RBP4) antagonist which is a non-retinol-like retinol binding protein 4 (RBP4) antagonist. The present invention provides a method for treating gout in a subject suffering from this disease, the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective for treating the subject, the compound being a retinol binding protein 4 (RBP4) antagonist.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 201980051205.3, titled "RBP4 Antagonists for the Treatment and Prevention of Non-Alcoholic Fatty Liver Disease and Gout", with a filing date of August 1, 2019. Technical Field

[0002] Throughout this application, certain publications are cited in parentheses. The complete citations of these publications can be found at the end of the specification. The disclosures of these publications are incorporated herein by reference in their entirety to more fully describe the prior art relevant to the present invention.

[0003] This invention was made with government support under Grant Nos. NS074476 and EY027027 awarded by the National Institutes of Health. The government has certain rights in this invention. Background Art

[0004] Non-alcoholic fatty liver disease (NAFLD)

[0005] Non-alcoholic fatty liver disease (NAFLD) encompasses a range of conditions related to lipid deposition in liver hepatocytes. Hepatic steatosis refers to the accumulation of lipids in the liver. NAFLD is characterized by hepatic steatosis caused by reasons other than excessive alcohol consumption. Clinically, hepatic steatosis is defined as a hepatic triglyceride content exceeding 5% of the total liver weight. Although simple hepatic steatosis is the least extreme within the NAFLD spectrum, it can progress to more severe conditions within the NAFLD spectrum, such as mild hepatic steatosis and non-alcoholic steatohepatitis (NASH). NASH is an extreme form of NAFLD, characterized by lipid accumulation in the liver, accompanied by inflammation and hepatocyte damage or fibrosis. NASH often leads to severe liver complications, such as cirrhosis and hepatocellular carcinoma.

[0006] NAFLD is the most common form of chronic liver disease in the United States, affecting an estimated 75 million to 100 million people. There is currently no approved drug therapy for any form of NAFLD. Developing a drug therapy for NAFLD is of great importance.

[0007] Gouty arthritis

[0008] Gouty arthritis (gout) is the most common form of inflammatory arthritis, affecting over 8 million people in the United States (Lawrence, R.C. et al. 2008). Uric acid is a metabolite produced by purine metabolism that is present in many foods and human tissues (Terkeltaub, R.A. 2001; Burns, C. et al. 2013). Gout is caused by elevated levels of uric acid in the blood, which lead to the deposition of monosodium urate crystals in tissues. These crystals form when the concentration of uric acid in tissues and in circulation exceeds the solubility limit, resulting in a gout attack. Risk factors for gout include being overweight or obese, having high blood pressure, drinking alcohol, using diuretics, having a diet rich in meat and seafood, consuming too much fructose, and having poor kidney function (Choi, H.K. et al. 2004a; Choi, H.K. 2004b; Krishnan, E. 2012).

[0009] An acute attack occurs when urate crystals in the joint cause acute inflammation. The attack is characterized by pain, redness, swelling, and heat that lasts from days to weeks. The pain can be mild or severe. Most initial attacks occur in the lower extremities. The typical presentation at the metatarsophalangeal joint of the big toe (podagra) is the presenting joint in 50% of gout patients. Chronic gout is characterized by chronic arthritis, with joint soreness and pain. People with gout may also develop tophi or nodules of urate crystals deposited in soft tissues. The clinically inactive (latent) phase between gout attacks occurs after the acute attack has subsided. People with gout continue to have hyperuricemia, which leads to the continued deposition of urate crystals in tissues and causes damage. As the disease progresses, the latent phase becomes shorter.

[0010] Uric acid is synthesized from its precursor xanthine by an enzyme called xanthine oxidase (XO). Therefore, XO inhibitors (such as allopurinol and febuxostat) are dominant on the market (Stamp, L.K. et al. 2015; Love, B.L. et al. 2010). However, the most common cause of elevated circulating uric acid levels is insufficient renal secretion of uric acid. The slightly more effective probenecid and the recently approved lesinurad are treatments that increase renal secretion of uric acid.

[0011] The incidence and prevalence of gout are rising. This is due to factors such as an increasing aging population and lifestyle factors, where many elderly people take thiazide diuretics and prophylactic aspirin that promote hyperuricemia, while lifestyle factors are characterized by excessive fructose and alcohol intake in the diet that promotes hyperuricemia, lack of physical activity, and abdominal fat accumulation (Burns, C. et al. 2013; Choi, H.K. et al. 2004a).

[0012] In the treatment of gout, there is still a large amount of unmet clinical need. Among 8 million gout patients, more than 3 million are receiving urate-lowering therapies (mainly XO inhibitors). Nevertheless, 1 million patients still experience 3 or more attacks each year, indicating the need for better urate-lowering therapies. SUMMARY OF THE INVENTION

[0013] The present invention provides a method for treating a subject suffering from non-alcoholic fatty liver disease (NAFLD), the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound is a non-retinoid retinol-binding protein 4 (RBP4) antagonist.

[0014] The present invention provides a method for treating a subject suffering from gout, the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound is a retinol-binding protein 4 (RBP4) antagonist.

[0015] The present invention provides a method for treating a subject suffering from non-alcoholic fatty liver disease (NAFLD) or gout, the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound has the following structure

[0016]

[0017] wherein L is a linking group having the following structure:

[0018]

[0019] and Z is a group having the following structure:

[0020]

[0021] wherein:

[0022] R1, R2, R3, R4 and R5 are each independently H, halogen, CF3 or C1-C4 alkyl;

[0023] R6 is H, OH or halogen or absent;

[0024] ψ is absent or present, and when present is a bond;

[0025] B is a substituted or unsubstituted heterobicyclic, pyridazine, pyrazole, pyrazine, thiadiazole or triazole, wherein the heterobicyclic is not a chlorine-substituted indole; and

[0026] When the pyrazole is substituted, it is not substituted with trifluoromethyl;

[0027] B’ is a substituted or unsubstituted phenyl, pyridine, pyrimidine, benzyl, pyrrolidine, sulfolane, oxetane, CO2H or (C1-C4 alkyl)-CO2H,

[0028] wherein the substituted phenyl is not substituted with trifluoromethyl or 3-(methyl carboxylate), the substituted pyridine is not substituted with trifluoromethyl, and the substituted pyrrolidine is not substituted with hydroxamic acid, and

[0029] the substituted or unsubstituted pyrrolidine is bonded to the carbonyl group through a carbon-carbon bond;

[0030] A is absent or present, and when present is B1 is a substituted or unsubstituted monocyclic, bicyclic, heteromonocyclic, heterobicyclic, benzyl, CO2H or (C1-C4 alkyl)-CO2H,

[0031] wherein when B1 is CO2H, then A is present and is R7 is an alkyl;

[0032] X is N or CR8, where R8 is H, OH or halogen;

[0033] B2 has the following structure:

[0034]

[0035] wherein:

[0036] α and β are each a bond that is present or absent;

[0037] X1 is N, NH or NR 99 ,

[0038] where R 99 is an alkyl, alkenyl or alkynyl;

[0039] X2 is C or N;

[0040] X3 is CH or N;

[0041] R9, R 10 and R 11 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3,

[0042] wherein:

[0043] X1, X2, and X3 are each N, α is present and β is absent; or

[0044] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or

[0045] X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0046] X1 is NH or NR 99 , X2 is C, X3 is N, α is absent and β is present, where when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R9, R 10 and R 11 is not H,

[0047] or B2 has the following structure:

[0048]

[0049] wherein:

[0050] R 12 、R 13 and R 14 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3,

[0051] or a pharmaceutically acceptable salt thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] Figure 1 : Single oral administration (PO) or intravenous administration (IV) of a dose of Compound 1 induces a strong reduction in the circulating levels of serum RBP4 in wild-type mice. The upper figure shows the data for intravenous administration of 2 mg of Compound 1 per kg of mouse body weight. The lower figure shows the data for oral administration of 5 mg of Compound 1 per kg of mouse body weight.

[0053]

[0054] Compound 1

[0055] Plasma samples were collected at baseline and 10 time points after compound 1 administration, and RBP4 levels were measured. Three groups of mice were used for oral and intravenous administration experiments (mouse group 1, mouse group 2, and mouse group 3). Each mouse group consisted of 5 animals. Plasma RBP4 levels were determined using an RBP4 (mouse / rat) DuoSet ELISA kit (AdipoGen, Switzerland) according to the manufacturer's instructions.

[0056] Figure 2 : As described by Lee 2016, an adi-hRBP4 transgenic mouse model was generated. The human hRBP4 transgene was introduced into the ROSA26 locus. The human transgene contained a loxP-flanked stop cassette that blocked expression. These mice were mated with mice expressing adiponectin-Cre, and thus the stop cassette was removed only in adipocytes. Therefore, in this model, human RBP4 was specifically expressed in adipose tissue.

[0057] Figure 3 : Experimental design for evaluating the efficacy of compound 1 in an adi-hRBP4 genetic model of hepatic steatosis.

[0058] Figure 4 : In the transgenic mouse line, compound 1 induced a decrease in serum levels of human and mouse RBP4. The left bar in each pair of bars is lightly shaded and represents baseline data. The right bar in each pair of bars is darkly shaded and represents the serum levels of human and mouse RBP4 at the end of the study.

[0059] Figure 5 : Dynamics of body weight changes in three groups of transgenic animals: Both groups on a high-fat diet (HFD) accumulated more weight than the control group. The body weight of the HFD group increased significantly compared to the standard diet group (*P<0.05; **P<0.01; two-way RM ANOVA, combined with Holm-Sidak post hoc comparison test)

[0060] Figure 6 : Significance between HFD and HFD + compound. Treatment with compound 1 significantly reduced the HFD-induced body weight gain at several time points. A significant difference in body weight was detected between the group with only HFD and the treatment group (HFD + compound 1) (*P<0.05; **P<0.01; two-way RM ANOVA, combined with Holm-Sidak post hoc comparison test).

[0061] Figure 7: As shown in the comparison between the treatment group (HFD + Compound 1) and the group with only HFD, treatment with Compound 1 significantly reduced body weight at the end of the 29-day treatment period. A significant difference was observed between the HFD group and the treatment group (HFD + Compound 1) (P = 0.0153; one-way RM ANOVA, combined with Holm-Sidak post hoc comparison test).

[0062] Figure 8 : There was no difference in food intake between the treatment (HFD + Compound 1) group and the HFD-only group (two-way RM ANOVA, combined with Holm-Sidak post hoc comparison test).

[0063] Figure 9 : Compound 1 significantly reduced hepatic free fatty acid concentration. At the end of the 29-day study, a significant difference was observed between the HFD-only group and the treatment group (HFD + Compound 1) (P = 0.011; one-way ANOVA, combined with Holm-Sidak post hoc comparison test). The data shown were determined at the end of the 29-day study.

[0064] Figure 10 : Compound 1 significantly reduced hepatic triglycerides. A significant difference was observed between the HFD-only group and the treatment group (HFD + Compound 1) (P = 0.01; one-way ANOVA, combined with Holm-Sidak post hoc comparison test). The data shown were determined at the end of the 29-day study.

[0065] Figure 11 : Compound 1 significantly reduced lipid deposition in the liver of treated animals. A. Liver histological score.

[0066] Figure 11 B: Compound 1 significantly reduced lipid deposition in the liver of treated animals. B. The average score for the normal diet group was 0.

[0067] Figure 11 C: Compound 1 significantly reduced lipid deposition in the liver of treated animals. C. The average score for the HFD-only group was 2.9.

[0068] Figure 11 D: Compound 1 significantly reduced lipid deposition in the liver of treated animals. D. The average score for the treatment group (HFD + Compound 1) was 1.6. A significant difference was observed between the HFD-only group and the treatment group (HFD + Compound 1) (P < 0.001; one-way ANOVA, combined with Holm-Sidak post hoc comparison test). The data shown were determined at the end of the 29-day study.

[0069] Figure 12: Compound 1 significantly reduced serum uric acid. There was a significant difference only between the HFD group and the treatment group (HFD + Compound 1) (P = 0.03; one-way ANOVA, combined with Holm-Sidak post hoc comparison test). However, there was no significant statistical difference between the normal diet group and the HFD-only group. The data shown were determined at the end of the 29-day study.

[0070] Figure 13 : No steatosis was observed in the kidneys of the animals under HFD. This figure is a representative image of the HFD-only group. No lipid droplets were observed in the renal tubules in any of the three experimental groups. The data shown were determined at the end of the 29-day study.

[0071] Figure 14 : Representative isotherms of A1120 (positive control) and fatty acids binding to human RBP4. 10 nM 3 H-retinol was used as the radioligand. Fatty acids (such as palmitic acid, oleic acid, and linoleic acid) could displace radiolabeled retinol from RBP4, indicating that they could bind to the retinol-binding pocket of RBP4. This data implies that RBP4 expressed in adipose tissue may be involved in the transport of fatty acids from adipose tissue to the liver. Compound 1 and other RBP4 antagonists can inhibit this fatty acid transport because they compete for binding to the same retinol-binding pocket in RBP4.

[0072]

[0073] Figure 15 : In vivo PK data of analog compounds 1 and 2 after IV and PO administration in rodents a 。

[0074] a The dosing groups consisted of three naive male CD-1 mice or adult male Sprague-Dawley rats. The data were presented as mean ± SD. b Total body clearance. c The maximum observed concentration of the compound in plasma. d The time of the maximum observed concentration of the compound in plasma after oral administration. e The apparent half-life at the terminal phase of elimination of the compound from plasma. f The volume of distribution at steady state. The area under the plasma concentration-time curve of the compound in plasma from 0 to the last time point was quantifiable. hBioavailability; F = (AUCINFpo × doseiv) ÷ (AUCINFiv × dosepo) Intravenous (IV) formulation = 3% dimethacrylate / 45% polyethylene glycol 300 / 12% ethanol / 40% sterile water; IV dosage = 2 mL / kg; Oral (PO) formulation = 2% Tween 80 in 0.9% saline; PO dosage = 5 mL / kg. Dosage regimens of Compound 3 in mice and rats: 2.0 mg / kg IV, 5 mg / kg PO. Dosage regimens of Compound 1 in mice: 2.0 mg / kg IV, 5 mg / kg PO. Dosage regimens of Compound 1 in rats: 1.0 mg / kg IV, 2.0 mg / kg IV, 5 mg / kg PO.

[0075] Figure 16 A PK / PD characteristics of Compound 1 in mice. Plasma RBP4 levels in CD-1 mice after single oral administration of 5 mg / kg of Compound 1.

[0076] Figure 16 B: PK / PD characteristics of Compound 1 in mice. Plasma RBP4 levels in CD-1 mice after single intravenous administration of 2 mg / kg of Compound 1.

[0077] Intravenous dose (D) of 2 mg / kg of Compound 1. Data are presented as mean ± SD. For each time point of blood collection, three mice were used in the study.

[0078] Figure 16 C: PK / PD characteristics of Compound 1 in mice. Plasma compound levels after single oral administration of a 5 mg / kg dose of Compound 1. Data are presented as mean ± SD. For each time point of blood collection, three mice were used in the study.

[0079] Figure 16 D: PK / PD characteristics of Compound 1 in mice. Plasma compound levels after single oral administration of a 2 mg / kg dose of Compound 1. Data are presented as mean ± SD. For each time point of blood collection, three mice were used in the study.

[0080] Figure 17Effect of oral administration of Compound 1 on circulating levels of serum RBP4 in adi-hRBP4 mice. Serum levels of murine RBP4 were measured at baseline (black circles) and at the end of 29-day compound treatment (red squares) using a species-specific rodent or human enzyme-linked immunosorbent assay. A statistically significant 90% decrease in both human and murine RBP4 was seen at the end of the study in Compound 1-treated mice compared to baseline (two-way ANOVA, combined with Holm-Sidak post-comparison test, ****P < 0.0001). A significant decrease in both human and murine RBP4 concentrations was detected in Compound 1-treated adi-hRBP4 mice compared to vehicle-treated knockout controls (two-way ANOVA, combined with Holm-Sidak post-comparison test, P < 0.0001). Error bars show SD; bar graphs show mean values. Each data point on the graph represents the serum RBP4 concentration from an individual animal. The number of male adi-hRBP4 mice in each treatment group was as follows: 8 on normal chow, 7 on HFD, and 8 on HFD with Compound 1.

[0081] Figure 17 B: Effect of oral administration of Compound 1 on circulating levels of serum RBP4 in adi-hRBP4 mice. Serum levels of human RBP4 were measured at baseline (black circles) and at the end of 29-day compound treatment (red squares) using a species-specific rodent or human enzyme-linked immunosorbent assay. A statistically significant 90% decrease in both human and murine RBP4 was seen at the end of the study in Compound 1-treated mice compared to baseline (two-way ANOVA, combined with Holm-Sidak post-comparison test, ****P < 0.0001). A significant decrease in both human and murine RBP4 concentrations was detected in Compound 1-treated adi-hRBP4 mice compared to vehicle-treated knockout controls (two-way ANOVA, combined with Holm-Sidak post-comparison test, P < 0.0001). Error bars show SD; bar graphs show mean values. Each data point on the graph represents the serum RBP4 concentration from an individual animal. The number of male adi-hRBP4 mice in each treatment group was as follows: 8 on normal chow, 7 on HFD, and 8 on HFD with Compound 1.

[0082] Figure 18Compound 1 partially prevented high-fat diet-induced obesity in adi-hRBP4 mice. Weight gain was measured in male adi-hRBP4 mice fed normal chow (n = 8), HFD (n = 7), and HFD with 58 (n = 8). Weight gain in compound-treated HFD mice was significantly reduced at four time points starting from day 19 compared with the untreated HFD group (two-way repeated measures (RM) ANOVA, combined with Holm-Sidak post hoc comparison test, *P < 0.05; **P < 0.01). At all time points studied, weight gain in food-fed mice was lower than that in the HFD group (two-way RM ANOVA, combined with Holm-Sidak post hoc comparison test, ***P < 0.001; ****P < 0.0001). Values represent mean percentage change in body weight relative to baseline. Error bars show SD.

[0083] Figure 18 B: Compound 1 partially prevented high-fat diet-induced obesity in adi-hRBP4 mice. Daily food consumption was normalized to body weight in male adi-hRBP4 mice fed normal chow, HFD, and HFD with compound 1. There was no difference in food consumption between the untreated HFD group and compound-treated HFD mice (two-way RM ANOVA, combined with Holm-Sidak post hoc comparison test). Values represent mean normalized daily food consumption. Error bars show SD.

[0084] Figure 19 A Effect of oral administration of compound 1 at a dose of 20 mg / kg on hepatic free fatty acid and triglyceride levels in obese adihRBP4 mice. Hepatic FFA (A) levels in male adi-hRBP4 mice fed normal chow (n = 8), HFD (n = 7), and HFD with compound 58 (n = 8). Hepatic FFA (P = 0.0107, one-way ANOVA, combined with Holm-Sidak post hoc comparison test) and triglyceride (P = 0.0104, one-way ANOVA, combined with Holm-Sidak post hoc comparison test) levels were significantly reduced in compound 1-treated HFD mice compared with the untreated HFD group. Graph bars show mean values; error bars show SD; *P < 0.05; ****P < 0.0001. Each data point on the graph represents FFA or TG concentration from an individual animal.

[0085] Figure 19B: Effect of oral administration of Compound 1 at a dose of 20 mg / kg on hepatic free fatty acid and triglyceride levels in obese adihRBP4 mice. Liver TG (B) levels in male adi-hRBP4 mice fed normal chow (n = 8), HFD (n = 7), and HFD with Compound 58 (n = 8). Compared with the untreated HFD group, hepatic FFA (P = 0.0107, one-way ANOVA, combined with Holm-Sidak post hoc comparison test) and triglyceride (P = 0.0104, one-way ANOVA, combined with Holm-Sidak post hoc comparison test) levels were significantly reduced in Compound 1-treated HFD mice. Graph bars show mean values; error bars show SD; *P < 0.05; ****P < 0.0001. Each data point on the graph represents the FFA or TG concentration from a single animal.

[0086] Figure 20 A Effect of Compound 1 on hepatic lipid deposition in adi-hRBP4 mice. Representative liver frozen sections stained with Oil Red O showed the fatty liver status in chow-fed, HFD, and Compound 1-treated HFD adi-hRBP4 mice. The compound was orally administered at a dose of 20 mg / kg.

[0087] Figure 20 B: Effect of Compound 1 on hepatic lipid deposition in adi-hRBP4 mice. Histological scores of Oil Red O-stained liver cryosections from chow-fed, HFD, and Compound 1-treated HFD adi-hRBP4 mice. Hepatic steatosis was classified as grade 0 (0% of hepatocytes had macrovesicular steatosis), grade 1 (<33% of hepatocytes had macrovesicular steatosis), grade 2 (33 - 66% of hepatocytes had macrovesicular steatosis), and grade 3 (>66% of hepatocytes had macrovesicular steatosis). Data were analyzed using one-way ANOVA (combined with Holm-Sidak post hoc test). Graph bars show mean values; error bars show SD; ***P < 0.001. Each data point on the graph represents the histological score of steatosis from a single animal. The number of male adi-hRBP4 mice in each treatment group was: normal chow 8, HFD 7, and HFD with Compound 1 8.

[0088] Figure 21: Binding of fatty acids to RBP4. (A) Isotherms of palmitic acid, oleic acid, linoleic acid, and docosahexaenoic acid binding to human RBP4. 10 nM of 3H-retinol was used as the radioligand. (B) Superposition of the minimum binding conformations of retinol (black, 5NU7), antagonist compound 3 (purple, 3FMZ), and palmitic acid, oleic acid, linoleic acid, and docosahexaenoic acid (orange). Phe36 is dark green in 3FMZ (adjacent to the fatty acid carboxylic acid group) and light green in the 5NU7 model (adjacent to the alcohol group of 1). Contact residues are labeled and illustrated in stick format (MOE, Chemical Computing Group, Inc., Montreal, Canada). Detailed Description

[0089] The present invention provides a method for treating a subject suffering from non-alcoholic fatty liver disease (NAFLD), the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound is a non-retinoid retinol-binding protein 4 (RBP4) antagonist.

[0090] The present invention provides a method for treating a subject suffering from gout, the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound is a retinol-binding protein 4 (RBP4) antagonist.

[0091] The present invention provides a method for treating a subject suffering from non-alcoholic fatty liver disease (NAFLD) or gout, the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound has the following structure

[0092]

[0093] wherein L is a linking group having the following structure:

[0094]

[0095] and Z is a group having the following structure:

[0096]

[0097] wherein:

[0098] R1, R2, R3, R4, and R5 are each independently H, halogen, CF3, or C1-C4 alkyl; R6 is H, OH, or halogen or is absent;

[0099] ψ is absent or present and, when present, is a bond;

[0100] B is a substituted or unsubstituted heterobicyclic, pyridazine, pyrazole, pyrazine, thiadiazole or triazole,

[0101] wherein said heterobicyclic is not a chlorine-substituted indole; and

[0102] when said pyrazole is substituted, it is not substituted with trifluoromethyl;

[0103] B’ is a substituted or unsubstituted phenyl, pyridine, pyrimidine, benzyl, pyrrolidine, sulfolane, oxetane, CO2H or (C1-C4 alkyl)-CO2H,

[0104] wherein said substituted phenyl is not substituted with trifluoromethyl or 3-(methyl carboxylate), said substituted pyridine is not substituted with trifluoromethyl, and said substituted pyrrolidine is not substituted with hydroxamic acid, and

[0105] said substituted or unsubstituted pyrrolidine is bonded to the carbonyl group through a carbon-carbon bond;

[0106] A is absent or present and, when present, is B1 is a substituted or unsubstituted monocyclic, bicyclic, heteromonocyclic, heterobicyclic, benzyl, CO2H or (C1-C4 alkyl)-CO2H,

[0107] wherein when B1 is CO2H, then A is present and is R7 is an alkyl;

[0108] X is N or CR8, where R8 is H, OH or a halogen;

[0109] B2 has the following structure:

[0110]

[0111] wherein:

[0112] α and β are each a bond that is present or absent;

[0113] X1 is N, NH or NR 99 ,

[0114] where R 99 is an alkyl, alkenyl or alkynyl;

[0115] X2 is C or N;

[0116] X3 is CH or N;

[0117] R9, R 10 and R 11each independently is H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl--NH2, alkyl-OAc, alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3,

[0118] wherein:

[0119] X1, X2 and X3 are each N, α is present and β is absent; or

[0120] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or

[0121] X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0122] X1 is NH or NR 99 , X2 is C, X3 is N, α is absent and β is present, wherein when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R9, R 10 and R 11 is not H,

[0123] or B2 has the following structure:

[0124]

[0125] wherein:

[0126] R 12 、R 13 and R 14 each independently is H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl--NH2, alkyl-Oac, alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3, or a pharmaceutically acceptable salt thereof.

[0127] In one embodiment, the subject has an NAFLD disease selected from the group consisting of: hepatic steatosis (fatty liver), non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma.

[0128] In some embodiments, the method further comprises the step of determining or having determined the level of RBP4 in the adipose tissue of a subject and administering the pharmaceutical composition if the level of RBP4 in the adipose tissue is elevated.

[0129] In one embodiment, the method further comprises the step of determining or having determined the level of RBP4 in the serum of a subject and administering the pharmaceutical composition if the level of RBP4 in the serum is elevated.

[0130] In one embodiment, the amount of the compound is effective to reduce the level of RBP4 in the adipose tissue of a subject. In another embodiment, the amount of the compound is effective to reduce the level of RBP4 in the serum of a subject. In another embodiment, the amount of the compound is effective to reduce the level of uric acid in the serum of a subject. In another embodiment, the amount of the compound is effective to normalize the concentration of triglycerides in the liver of a subject.

[0131] In one embodiment, the amount of the compound is effective to normalize the concentration of free fatty acids in the serum of a subject. In another embodiment, the amount of the compound is effective to normalize the concentration of free fatty acids in the liver of a subject. In some embodiments, the amount of the compound is effective to prevent the transport of fatty acids by RBP4. In further embodiments, the amount of the compound is effective to prevent the transport of fatty acids by RBP4 to the liver. In one embodiment, the amount of the compound is effective to inhibit the binding between RBP4 and fatty acids.

[0132] In one embodiment, the fatty acid is from adipose tissue.

[0133] In one embodiment, the subject does not have an elevated serum RBP4 level. In another embodiment, the subject has an elevated serum RBP4 level. In some embodiments, the serum RBP4 level is elevated by more than 3 micrograms / ml.

[0134] In some embodiments, the NAFLD is hepatic steatosis selected from simple hepatic steatosis and mild hepatic steatosis.

[0135] In one embodiment, the compound is not a ligand of the nuclear receptor RAR. In another embodiment, the RBP4 antagonist is a non-retinoid antagonist. In another embodiment, the RBP4 antagonist is not fenritinide.

[0136] In some embodiments, L is and Z is

[0137] In further embodiments, L is and Z is In additional embodiments, L is Z is Two or more of R1, R2, R3, R4, or R5 are not H,

[0138] and when R1 is CF3, R2 is H, R3 is F, R4 is H, and R5 is H, or R1 is H, R2 is CF3, R3 is H, R4 is CF3, and R5 is H, or R1 is Cl, R2 is H, R3 is H, R4 is F, and R5 is H, or R1 is CF3, R2 is H, R3 is F, R4 is H, and R5 is H, or R1 is CF3, R2 is F, R3 is H, R4 is H, and R5 is H, or R1 is Cl, R2 is F, R3 is H, R4 is H, and R5 is H, then B is not

[0139]

[0140] In some embodiments, L is and Z is is Z is and

[0141] R6 is absent or present, and when present is H, OH, or

[0142] halogen and when ψ is present, then R6 is absent, and when ψ is absent, then R6 is present.

[0143] In some embodiments, L is and

[0144] Z is

[0145] In some embodiments, L is and

[0146] Z is

[0147] In some embodiments, L is Z is R6 is H, and A is

[0148] In some embodiments, L is and Z is

[0149] In some embodiments, the compound is:

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171]

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183] or a pharmaceutically acceptable salt thereof.

[0184] In some embodiments, the compound is:

[0185]

[0186]

[0187]

[0188]

[0189] or a pharmaceutically acceptable salt thereof.

[0190] In some embodiments, the compound is:

[0191]

[0192]

[0193]

[0194]

[0195] or a pharmaceutically acceptable salt thereof.

[0196] In some embodiments, the compound is:

[0197]

[0198]

[0199]

[0200] or a pharmaceutically acceptable salt thereof.

[0201] In some embodiments, the compound is:

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208] or a pharmaceutically acceptable salt thereof.

[0209] In some embodiments, the compound is:

[0210]

[0211]

[0212] or a pharmaceutically acceptable salt thereof.

[0213] In some embodiments, the compound is:

[0214]

[0215]

[0216]

[0217] or a pharmaceutically acceptable salt thereof.

[0218] In some embodiments, the compound is:

[0219] or a pharmaceutically acceptable salt thereof.

[0220] In some embodiments, the compound is:

[0221]

[0222]

[0223] or a pharmaceutically acceptable salt thereof.

[0224] The method of the present invention includes a pharmaceutical composition, wherein the compound has the following structure:

[0225]

[0226] Wherein:

[0227] R1, R2, R3, R4 and R5 are each independently H, halogen, CF3 or C1-C4 alkyl; X is N or CR6,

[0228] wherein R6 is H, OH or halogen;

[0229] A is absent or present, and when present is B has the following structure:

[0230]

[0231] wherein:

[0232] α and β are each a bond that is present or absent;

[0233] X1 is N, NH or NR 10 ,

[0234] where R 10 is alkyl, alkenyl or alkynyl;

[0235] X2 is C or N;

[0236] X3 is CH or N;

[0237] R7, R8 and R9 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl--NH2, alkyl-OAc alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3,

[0238] wherein:

[0239] X1, X2 and X3 are each N, α is present and β is absent; or

[0240] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or

[0241] X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0242] X1 is NH or NR 10 , X2 is C, X3 is N, α is absent and β is present, wherein when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R7, R8 and R9 is not H,

[0243] Alternatively, B has the following structure:

[0244]

[0245] Wherein:

[0246] R 11 、R 12 and R 13 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3, or a pharmaceutically acceptable salt thereof.

[0247] In some embodiments, the compounds of the method have the following structure:

[0248]

[0249] In some embodiments, the compounds of the method have the following structure:

[0250]

[0251] In some embodiments, the compounds of the method have the following structure:

[0252]

[0253] In some embodiments, the compounds of the method have the following structure:

[0254]

[0255] Wherein:

[0256] R1, R2, R3, R4 and R5 are each independently H, halogen, CF3 or C1-C4 alkyl; and B has the following structure:

[0257]

[0258] Wherein:

[0259] α and β are each a bond that is present or absent;

[0260] X1 is N, NH or NR 10 ,

[0261] where R 10 is alkyl, alkenyl or alkynyl;

[0262] X2 is C or N;

[0263] X3 is CH or N;

[0264] R7, R8, and R9 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN, or CF3, where:

[0265] X1, X2, and X3 are each N, α is present and β is absent; or

[0266] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or

[0267] X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0268] X1 is NH or NR 10 、X2 is C, X3 is N, α is absent and β is present, where when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R7, R8, and R9 is not H,

[0269] Alternatively, B has the following structure:

[0270]

[0271] where:

[0272] R 11 、R 12 and R 13 are each independently H, halogen, alkyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN, or CF3,

[0273] or a pharmaceutically acceptable salt thereof.

[0274] In some embodiments, the compound of the method has the following structure:

[0275]

[0276] where:

[0277] R1, R2, R3, R4, and R5 are each independently H, halogen, CF3, or C1-C4 alkyl;

[0278] Y is alkyl;

[0279] A is either absent or present, and when present, is and B has the following structure:

[0280]

[0281] wherein:

[0282] α and β are each a bond that is either present or absent;

[0283] X1 is N, NH, or NR 10 ,

[0284] where R 10 is alkyl, alkenyl, or alkynyl;

[0285] X2 is C or N;

[0286] X3 is CH or N;

[0287] R7, R8, and R9 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN, or CF3, wherein:

[0288] X1, X2, and X3 are each N, α is present and β is absent; or

[0289] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or

[0290] X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0291] X1 is NH or NR 10 , X2 is C, X3 is N, α is absent and β is present, where when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R7, R8, and R9 is not H,

[0292] Alternatively, B has the following structure:

[0293]

[0294] wherein:

[0295] R 11 , R 12 and R 13Each independently is H, halogen, alkyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3,

[0296] or a pharmaceutically acceptable salt thereof.

[0297] In some embodiments, the compounds of the method have the following structure:

[0298]

[0299] In some embodiments, the compounds of the method have the following structure:

[0300]

[0301] Wherein:

[0302] α and β are each a bond that is present or absent;

[0303] X1 is N, NH or NR 10 ,

[0304] wherein R 10 is alkyl, alkenyl or alkynyl;

[0305] X2 is C or N;

[0306] X3 is CH or N;

[0307] R7, R8 and R9 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl--NH2, alkyl-OAc alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3,

[0308] Wherein:

[0309] X1, X2 and X3 are each N, α is present and β is absent; or

[0310] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or

[0311] X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0312] X1 is NH or NR 10, X2 is C, X3 is N, α is absent and β is present, where when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R7, R8, and R9 is not H.

[0313] In some embodiments, B or B2 has the following structure:

[0314]

[0315] Wherein:

[0316] R 11 、R 12 and R 13 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3.

[0317] In some embodiments, B or B2 has the following structure:

[0318]

[0319] R7, R8, and R9 are each independently H, halogen, alkyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3.

[0320] In some embodiments, B or B2 has the following structure:

[0321]

[0322] R7, R8, and R9 are each independently H, halogen, alkyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3.

[0323] In some embodiments, B or B2 has the following structure:

[0324]

[0325] R7, R8, and R9 are each independently H, halogen, alkyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN, or CF3.

[0326] In some embodiments, B or B2 has the following structure:

[0327]

[0328] R7, R8, and R9 are each independently H, halogen, alkyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN, or CF3; and R 10 is alkyl, alkenyl, or alkynyl.

[0329] In some embodiments, R7, R8, and R9 are each independently H, Cl, Br, F, OCH3, OCH2CH3, CF3, CN, CH3, CH3CH3, C(O)OH, or C(O)-NH2.

[0330] In some embodiments, R7, R8, and R9 are each independently H, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0331] In some embodiments, R7, R8, and R9 are each independently H, halogen, or alkyl.

[0332] In some embodiments, two of R7, R8, and R9 are both H, and the remaining one of R7, R8, and R9 is not H.

[0333] In some embodiments, one of R7, R8, and R9 is H, and the remaining two of R7, R8, and R9 are both not H.

[0334] In some embodiments, B or B2 has the following structure:

[0335]

[0336] In some embodiments, R7, R8, and R9 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0337] In some embodiments, B or B2 has the following structure:

[0338]

[0339] In some embodiments, R7 and R9 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0340] In some embodiments, B or B2 has the following structure:

[0341]

[0342] In some embodiments, R7, R8, and R9 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0343] In some embodiments, B or B2 has the following structure:

[0344]

[0345] In some embodiments, R7 and R9 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0346] In some embodiments, B in the compound has the following structure:

[0347]

[0348] In some embodiments, R7, R8, and R9 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0349] In some embodiments, B or B2 has the following structure:

[0350]

[0351] In some embodiments, R7 and R9 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F or CH2CH2Br.

[0352] In some embodiments, B or B2 has the following structure:

[0353]

[0354] In some embodiments, R7, R8 and R9 in the compound are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F or CH2CH2Br; and R 10 is an alkyl group.

[0355] In some embodiments, B or B2 has the following structure:

[0356]

[0357] In some embodiments, R7 and R9 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F or CH2CH2Br; and R 10 is an alkyl group.

[0358] In some embodiments, B or B2 has the following structure:

[0359]

[0360] R 11 、R 12 and R 13 are each independently H, a halogen, an alkyl group, an alkyl-OH, an alkyl-NH2, an alkyl-OAc, an alkyl-O-alkyl, a haloalkyl group, a cycloalkyl group, an O-alkyl, an NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3.

[0361] In some embodiments, R 11 、R 12 and R 13 are each independently H, Cl, Br, F, OCH3, OCH2CH3, CF3, CN, CH3, CH3CH3, C(O)OH or C(O)-NH2.

[0362] In some embodiments, R 11 , R 12 and R 13 are each independently H, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0363] In some embodiments, R 11 , R 12 and R 13 are each independently H, a halogen, or an alkyl group.

[0364] In some embodiments, two of R 11 , R 12 and R 13 are H, while the remaining one of R 11 , R 12 and R 13 is not H.

[0365] In some embodiments, one of R 11 , R 12 and R 13 is H, while the remaining two of R 11 , R 12 and R 13 are not H.

[0366] In some embodiments, B or B2 has the following structure:

[0367]

[0368] In some embodiments, R 11 , R 12 and R 13 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0369] In some embodiments, B or B2 has the following structure:

[0370]

[0371] In some embodiments, R 11 and R 13 are each independently H, CH3, Br, Cl, F, CH2CH2OH, CH2CH2OCH3, CH2CH2OAc, CH2CH2Cl, CH2CH2F, or CH2CH2Br.

[0372] In some embodiments, X is N. In some embodiments, X of the compound is CH.

[0373] In some embodiments, R1, R2, R3, R4, and R5 are each H, t-Bu, Cl, F, or CF3.

[0374] In some embodiments, R1, R2, R3, and R4 are each H; and

[0375] R5 is CF3 or t-Bu.

[0376] In some embodiments,

[0377] R1, R3, and R4 are each H;

[0378] R2 is a halogen;

[0379] R5 is CF3 or t-Bu.

[0380] In some embodiments,

[0381] R1, R2, R3, and R4 are each H;

[0382] R5 is CF3 or t-Bu.

[0383] In some embodiments,

[0384] R1, R2, R3, and R4 are each H;

[0385] R5 is CF3.

[0386] In some embodiments, one of R1, R2, R3, R4, and R5 is not H.

[0387] In some embodiments, two of R1, R2, R3, R4, and R5 are not H.

[0388] In some embodiments, two or more of R1, R2, R3, R4, and R5 are not H.

[0389] In some embodiments, three of R1, R2, R3, R4, and R5 are not H.

[0390] In some embodiments, three or more of R1, R2, R3, R4, and R5 are not H.

[0391] In some embodiments, the compound of the method has the following structure:

[0392]

[0393]

[0394]

[0395] Its pharmaceutically acceptable salts.

[0396] In some embodiments, B or B2 is not

[0397] The present invention provides compounds having the following structure:

[0398]

[0399] Wherein:

[0400] R1, R2, R3, R4 and R5 are each independently H, halogen, alkyl, haloalkyl, O-haloalkyl, aryl or heteroaryl;

[0401] X is N or CR6,

[0402] wherein R6 is H, OH or halogen;

[0403] A is absent or present, and when present is B has the following structure:

[0404]

[0405] Wherein:

[0406] α and β are each a bond that is present or absent;

[0407] X1 is N, NH or NR 10 ,

[0408] wherein R 10 is alkyl, alkenyl or alkynyl;

[0409] X2 is C or N;

[0410] X3 is CH or N;

[0411] R7, R8 and R9 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl--NH2, alkyl-OAc alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3,

[0412] Wherein:

[0413] X1, X2 and X3 are each N, α is present and β is absent; or

[0414] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or

[0415] X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0416] X1 is NH or NR 10 , X2 is C, X3 is N, α is absent and β is present, wherein when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R7, R8 and R9 is not H, or, B has the following structure:

[0417]

[0418] wherein

[0419] X4 and X5 are each independently N or CH; and

[0420] R 11 , R 12 and R 13 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3, or a pharmaceutically acceptable salt thereof.

[0421] 54. A compound having the following structure:

[0422]

[0423] wherein:

[0424] R1, R2, R3, R4 and R5 are each independently H, halogen, CF3 or C1-C4 alkyl, aryl or heteroaryl; and

[0425] B has the following structure:

[0426]

[0427] wherein:

[0428] α and β are each a bond that is present or absent;

[0429] X1 is N, NH or NR 10 ,

[0430] wherein R 10 is alkyl, alkenyl or alkynyl;

[0431] X2 is C or N;

[0432] X3 is CH or N;

[0433] R7, R8, and R9 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN, or CF3, where:

[0434] X1, X2, and X3 are each N, α is present and β is absent; or

[0435] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0436] X1 is NH or NR 10 , X2 is C, X3 is N, α is absent and β is present, where when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R7, R8, and R9 is not H,

[0437] Or, B has the following structure:

[0438]

[0439] Where:

[0440] R 11 , R 12 and R 13 are each independently H, halogen, alkyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN, or CF3,

[0441] or a pharmaceutically acceptable salt thereof.

[0442] A compound having the following structure:

[0443]

[0444] Where:

[0445] R1, R2, R3, R4, and R5 are each independently H, halogen, CF3, or a C1-C4 alkyl, aryl, or heteroaryl; Y is alkyl;

[0446] A is absent or present, and when present, is and B has the following structure:

[0447]

[0448] wherein:

[0449] α and β are each a bond that is present or absent;

[0450] X1 is N, NH or NR 10 ,

[0451] where R 10 is alkyl, alkenyl or alkynyl;

[0452] X2 is C or N;

[0453] X3 is CH or N;

[0454] R7, R8 and R9 are each independently H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3, wherein:

[0455] X1, X2 and X3 are each N, α is present and β is absent; or

[0456] X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or X1 is N, X2 is N, X3 is CH, α is present and β is absent; or

[0457] X1 is NH or NR 10 , X2 is C, X3 is N, α is absent and β is present, where when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R7, R8 and R9 is not H,

[0458] Alternatively, B has the following structure:

[0459]

[0460] where

[0461] R 11 , R 12 and R 13each independently is H, halogen, alkyl, alkyl-OH, alkyl-NH2, alkyl-OAc, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3,

[0462] or a pharmaceutically acceptable salt thereof.

[0463] In some embodiments of the above compounds, one of R1, R2, R3, R4 and R5 is not H.

[0464] In some embodiments of the above compounds, two of R1, R2, R3, R4 and R5 are not H.

[0465] In some embodiments of the above compounds, R1, R2, R3, R4 and R5 are H, methyl, ethyl, phenyl, t-Bu, i-Pr, OCF3, CF3, OCF2CF3, CF2CF3, Cl, Br or F, respectively.

[0466] In some embodiments of the above compounds, R1, R2, R3 and R4 are all H; and R5 is -H, OCF3, CF2CF3, methyl, ethyl, i-Pr or phenyl.

[0467] In some embodiments, the compound has the following structure:

[0468]

[0469] its pharmaceutically acceptable salt.

[0470] The compounds and classes of compounds disclosed in PCT International Publication Nos. WO 2014 / 152013, WO 2015 / 168286, WO 2014 / 151959, WO 2014 / 152018 and WO 2014 / 151936 can be used in the methods of the present invention. Accordingly, the compounds and classes of compounds disclosed in each of PCT International Publication Nos. WO 2014 / 152013, WO 2015 / 168286, WO 2014 / 151959, WO 2014 / 152018 and WO 2014 / 151936 are hereby incorporated by reference into the methods of the present invention.

[0471] In an embodiment, the compound is an RBP4 antagonist.

[0472] In one embodiment, the amount of the compound is 5 - 1000 mg, 5 - 800 mg, 5 - 200 mg, 45 - 200 mg, 45 - 1000 mg, 45 - 800 mg, 10 - 50 mg, 96 mg, 24 mg, or 10 mg per day.

[0473] In some embodiments, the method further comprises administering an amount of a second agent, the second agent being (R)-(+)-[(5,6-dichloro-2,3,9,9a-tetrahydro-3-oxo-9a-propyl-1H-fluoren-7-yl)oxy]acetic acid (DPOFA), a non-steroidal anti-inflammatory drug (NSAID) (such as indomethacin, colchicine, lesinurad), a corticosteroid (e.g., betamethasone, prednisone, dexamethasone, cortisone, hydrocortisone, methylprednisolone, prednisolone), a biologic anti-IL-1alpha / beta agent (e.g., canakinumab, rilonacept, anakinra), allopurinol, benzbromarone, pegloticase, and other forms of uricase, topiroxostat (FYX-051), ulodesine (BCX4208), KUX-1151, RLBN1001, RDEA3170, alprofenac (MBX-102), levotofisopam, UR-1102, PF-06743649, BCX4208, SHR4640, lumiracoxib, tranilast, topiroxostat, LC350189, bucillamine, AC-201, Huzhen capsules (including Polygonum cuspidatum and Ligustrum lucidum), MPC-004, FYU-981, sodium bicarbonate, SEL-212, SEL-037, apremilast, TMX-67, SSS11, D-0120, febuxostat, or probenecid, or an ester or salt thereof effective in treating a subject, thereby treating the subject.

[0474] In one embodiment, the second agent is DPOFA.

[0475] In some embodiments, the subject has gout.

[0476] In one embodiment, the amount of the second agent or the amount of the compound is effective to reduce the uric acid level in the blood of the subject. In another embodiment, the amount of the second agent or the amount of the compound is effective to reduce the uric acid reabsorption in the kidneys of the subject.

[0477] In one embodiment, the amount of the second agent can effectively increase the uric acid clearance rate in a subject.

[0478] In some embodiments, the amount of the second agent can effectively increase the uric acid level in the urine of a subject.

[0479] In one embodiment, the amount of the second agent can effectively increase the renal clearance rate of uric acid in a subject. In additional embodiments, the amount of the second agent or the amount of the compound can effectively alleviate one or more symptoms associated with gout in a subject.

[0480] In some embodiments, the one or more symptoms associated with gout are joint pain, arthritis, joint swelling and redness, and reduced range of motion of the joint. In additional embodiments, the amount of the second agent or the amount of the compound can effectively prevent gout in a subject.

[0481] In one embodiment, the prevention includes increasing the uric acid level in the urine of a subject. In another embodiment, the prevention includes reducing the uric acid level in the blood of a subject. In another embodiment, the prevention includes increasing the uric acid clearance rate of a subject. In another embodiment, the prevention includes reducing the reabsorption of uric acid in the kidneys of a subject.

[0482] In one embodiment, the prevention includes increasing the renal clearance rate of uric acid in a subject. In another embodiment, the prevention includes alleviating one or more symptoms associated with gout in a subject.

[0483] In some embodiments, the one or more symptoms associated with gout are joint pain, arthritis, joint swelling and redness, and reduced range of motion of the joint.

[0484] In one embodiment, the gout is chronic gout. In different embodiments, the gout is acute gout.

[0485] In some embodiments, the amount of the second agent or the amount of the compound prevents recurrence of chronic gout.

[0486] In one embodiment of the present invention, the subject is a mammal.

[0487] In some embodiments, the subject is female, and administration of the compound reduces the uric acid level by 2.4 - 6.0 mg / dL. In another embodiment, the subject is male, and administration of the compound reduces the uric acid level by 3.4 - 7.0 mg / dL.

[0488] In another embodiment, administration of the compound reduces the uric acid level in the subject to less than 7 mg / dL.

[0489] The present invention also provides a pharmaceutical composition for treating a subject suffering from non - alcoholic fatty liver disease (NAFLD) or gout, said pharmaceutical composition comprising an amount of a retinol - binding protein 4 (RBP4) antagonist or a compound as defined above.

[0490] In some embodiments, the pharmaceutical composition further comprises an amount of a second agent for treating a subject suffering from non - alcoholic fatty liver disease (NAFLD) or gout.

[0491] In some embodiments, the RBP4 antagonist or compound and the second agent are prepared for simultaneous, contemporaneous or combined administration.

[0492] The present invention further provides a pharmaceutical composition comprising a retinol - binding protein 4 (RBP4) antagonist or a compound as defined above, said pharmaceutical composition being used in combination therapy with a pharmaceutical composition comprising a second drug for treating non - alcoholic fatty liver disease (NAFLD) or gout.

[0493] The present invention additionally provides a pharmaceutical composition comprising an amount of a retinol - binding protein 4 (RBP4) antagonist or a compound as defined above, as adjunctive therapy to a second agent or in combination with a second drug, for treating a subject suffering from non - alcoholic fatty liver disease (NAFLD) or gout.

[0494] The present invention also provides the use of a retinol - binding protein 4 (RBP4) antagonist or a compound as defined above in the preparation of a medicament for treating a subject suffering from non - alcoholic fatty liver disease (NAFLD) or gout.

[0495] As used herein, "combination" refers to a combination of agents for use in therapy by simultaneous or contemporaneous administration. Simultaneous administration refers to the administration of a mixture of a first compound and a second compound (whether a true mixture, suspension, emulsion or other physical combination). In this case, the combination can be a mixture of the first compound and the second compound combined just prior to administration or in separate containers. Contemporaneous administration refers to the administration of the first compound and the second compound either simultaneously or at sufficiently close times such that additive and / or preferably synergistic activity relative to the activity of the first compound or the second compound alone is observed.

[0496] As used herein, "co - administration" or "co - administered" refers to the administration of two agents given at sufficiently close time intervals to allow the separate therapeutic effects of each agent to overlap.

[0497] As used herein, "add-on" or "add-on therapy" refers to a combination of agents for a therapy, where a subject receiving the therapy initiates a first treatment regimen of one or more agents and, before initiating a second treatment regimen of one or more different agents other than the first treatment regimen, whereby not all of the agents used in the therapy are initiated simultaneously. For example, adding Compound 1 therapy to a subject who has already received DPOFA therapy.

[0498] As used herein, a "non-retinoid RBP4 antagonist" is an RBP4 antagonist that is not a retinoid. A retinoid is a natural or synthetic analogue of retinol, which consists of four isoprenoid units linked head-to-tail. Retinoids are described in the Joint Commission on Biochemical Nomenclature of IUPAC-IUB (1982).

[0499] Unless otherwise stated, when the structure of a compound of the invention includes an asymmetric carbon atom, it is understood that the compound exists as a racemate, a racemic mixture, and as isolated single enantiomers. All such isomeric forms of these compounds are expressly included in the invention. Unless otherwise stated, each stereoisomeric carbon can be of the R or S configuration. It is thus understood that, unless otherwise stated, isomers arising from such asymmetry (e.g., all enantiomers and diastereomers) are included within the scope of the invention. Such isomers can be obtained in substantially pure form by classical separation techniques and by stereochemically controlled synthesis (as described in "Enantiomers, Racemates and Resolutions" by J. Jacques, A. Collet, and S. Wilen, published by John Wiley & Sons, NY, 1981). For example, separation can be carried out by preparative chromatography on a chiral column.

[0500] The invention also intends to include all isotopes of atoms that occur on the compounds disclosed herein. Isotopes include those atoms having the same atomic number but different mass numbers. As general examples and not as limitations, isotopes of hydrogen include tritium and deuterium. Isotopes of carbon include C-13 and C-14.

[0501] It should be noted that any symbol for carbon in the structures throughout this application, when no other symbol is used, is intended to represent all isotopes of carbon, e.g., 12 C, 13 C or 14 C. Further, any compound containing 13 C or 14 C can specifically have the structure of any compound disclosed herein.

[0502] It should be noted that any symbol for hydrogen in the structures throughout this application, when no other symbol is used, is intended to represent all isotopes of hydrogen, such as 1 H, 2 H or 3 H. In addition, any compound containing 2 H or 3 H can specifically have the structure of any compound disclosed herein.

[0503] Isotopically labeled compounds can generally be prepared by conventional techniques known to those skilled in the art, using appropriate isotopically labeled reagents in place of the non-labeled reagents employed.

[0504] The terms "substituted", "substituent", and "substitution" refer to the functional groups as described above, where one or more bonds to the hydrogen atoms contained therein are replaced by bonds to non-hydrogen or non-carbon atoms, provided that normal valences are maintained and the substitution results in a stable compound. Substituted groups also include groups where one or more bonds to one or more carbon or one or more hydrogen atoms are replaced by one or more bonds to heteroatoms (including double or triple bonds). Examples of substituents include the above-described functional groups and halogens (i.e., F, Cl, Br, and I); alkyl groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, and trifluoromethyl; hydroxy; alkoxy groups such as methoxy, ethoxy, n-propoxy, and isopropoxy; aryloxy groups such as phenoxy; arylalkoxy groups such as benzyloxy (phenylmethoxy) and p-trifluoromethylbenzyloxy (4-trifluoromethylphenylmethoxy); heteroaryloxy; sulfonyl groups such as trifluoromethanesulfonyl, methanesulfonyl, and p-toluenesulfonyl; nitro, nitrosyl; mercapto; thioalkyl groups such as methylthio, ethylthio, and propylthio; cyano; amino groups such as amino, methylamino, dimethylamino, ethylamino, and diethylamino; and carboxyl. When multiple substituent moieties are disclosed or claimed, the substituted compound can be substituted singly or multiply by one or more of the disclosed or claimed substituent moieties independently. Independent substitution means that the (two or more) substituents can be the same or different.

[0505] In the compounds used in the methods of the present invention, unless otherwise specifically defined, the substituents can be substituted or unsubstituted.

[0506] In the compounds used in the methods of the present invention, alkyl, heteroalkyl, monocyclic, bicyclic, aryl, heteroaryl, and heterocyclic groups can be further substituted by replacing one or more hydrogen atoms with alternative non-hydrogen groups. These include, but are not limited to, halogens, hydroxy, mercapto, amino, carboxyl, cyano, and carbamoyl.

[0507] It will be understood that the substituents and the substitution patterns on the compounds used in the method of the present invention can be selected by those of ordinary skill in the art to provide compounds that are chemically stable and can be readily synthesized from readily available starting materials by techniques known in the art. If the substituent itself is substituted by more than one group, it should be understood that these multiple groups can be on the same carbon or on different carbons, as long as a stable structure is obtained.

[0508] In selecting the compounds for use in the method of the present invention, those of ordinary skill in the art will recognize that the various substituents, i.e., R1, R2, etc., will be selected in accordance with well-known principles of chemical structure connectivity.

[0509] As used herein, "alkyl" is intended to include branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. Thus, as in "C1-C n alkyl", C1-C n is defined to include groups having a linear or branched arrangement of 1, 2,......, n-1 or n carbons, and specifically includes methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, isopropyl, isobutyl, sec-butyl, etc. One embodiment can be C1-C 12 alkyl, C2-C 12 alkyl, C3-C 12 alkyl, C4-C 12 alkyl, etc. One embodiment can be C1-C8 alkyl, C2-C8 alkyl, C3-C8 alkyl, C4-C8 alkyl, etc. "Alkoxy" denotes an alkyl group as described above linked by an oxygen bridge.

[0510] The term "alkenyl" refers to a straight-chain or branched non-aromatic hydrocarbon group that contains at least 1 carbon-carbon double bond, and up to the maximum number of non-aromatic carbon-carbon double bonds that can be present. Thus, C2-C n alkenyl is defined to include groups having 1, 2,......, n-1 or n carbons. For example, "C2-C6 alkenyl" refers to an alkenyl group that has 2, 3, 4, 5 or 6 carbon atoms, and respectively at least one carbon-carbon double bond and up to 3 carbon-carbon double bonds in the case of C6 alkenyl. Alkenyl includes vinyl, propenyl, butenyl, and cyclohexenyl. As described above for alkyl, the straight-chain, branched, or cyclic portion of the alkenyl can contain double bonds and can be substituted if a substituted alkenyl is indicated. One embodiment can be C2-C 12 alkenyl or C2-C8 alkenyl.

[0511] The term "alkynyl" refers to a straight-chain or branched non-aromatic hydrocarbon group that contains at least 1 carbon-carbon triple bond, and up to the maximum number of non-aromatic carbon-carbon triple bonds that can be present. Thus, C2-C nAlkynyl is defined to include groups having 1, 2,......, n - 1 or n carbons. For example, "C2 - C6 alkynyl" refers to an alkynyl group having 2 or 3 carbon atoms and 1 carbon - carbon triple bond, or an alkynyl group having 4 or 5 carbon atoms and at most 2 carbon - carbon triple bonds, or an alkynyl group having 6 carbon atoms and at most 3 carbon - carbon triple bonds. Alkynyl includes ethynyl, propynyl, and butynyl. As described above for alkyl, the straight - chain or branched - chain portion of the alkynyl may contain a triple bond and may be substituted if a substituted alkynyl is indicated. One embodiment may be C2 - C n alkynyl. One embodiment may be C2 - C 12 alkynyl or C3 - C8 alkynyl.

[0512] Alkyl may be unsubstituted or substituted by one or more substituents, including but not limited to halogen, alkoxy, alkylthio, trifluoromethyl, difluoromethyl, methoxy, and hydroxy.

[0513] As used herein, "C1 - C4 alkyl" includes branched and straight - chain C1 - C4 alkyl.

[0514] As used herein, "heteroalkyl" includes branched and straight - chain saturated aliphatic hydrocarbon groups having at least 1 heteroatom within the chain or branch.

[0515] As used herein, "cycloalkyl" includes rings of alkanes having a total of three to eight carbon atoms or any number within that range (i.e., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl).

[0516] As used herein, "heterocycloalkyl" is intended to mean a 5 - to 10 - membered non - aromatic ring containing 1 to 4 heteroatoms selected from O, N, and S, and includes bicyclic groups. Thus, "heterocyclic group" includes but is not limited to the following: imidazolyl, piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, tetrahydropyranyl, dihydropiperidinyl, tetrahydrothienyl, etc. If the heterocycle contains nitrogen, it is understood that its corresponding N - oxide is also covered by this definition.

[0517] As used herein, "aryl" is intended to mean any stable monocyclic, bicyclic, or polycyclic carbocyclic ring having at most 10 atoms in each ring, where at least one ring is aromatic and may be unsubstituted or substituted. Examples of such aryl moieties include but are not limited to: phenyl, p - tolyl (4 - methylphenyl), naphthyl, tetrahydronaphthyl, indanyl, phenanthryl, anthryl, or acenaphthylenyl. In the case where the aryl substituent is bicyclic and one ring is non - aromatic, it is understood that the attachment is through the aromatic ring.

[0518] The term "alkylaryl" refers to an alkyl group as described above, wherein one or more bonds to the hydrogen contained therein are replaced by bonds to an aryl group as described above. It should be understood that the "alkylaryl" group is attached to the core molecule through a bond from the alkyl group, and the aryl group serves as a substituent on the alkyl group. Examples of the arylalkyl moiety include, but are not limited to, benzyl (phenylmethyl), p-trifluoromethylbenzyl (4-trifluoromethylphenylmethyl), 1-phenylethyl, 2-phenylethyl, 3-phenylpropyl, 2-phenylpropyl, and the like.

[0519] As used herein, the term "heteroaryl" refers to a stable monocyclic, bicyclic or polycyclic ring having up to 10 atoms in each ring, wherein at least one ring is aromatic and contains 1-4 heteroatoms selected from O, N and S. Bicyclic aromatic heteroaryls include, but are not limited to, phenyl, pyridine, pyrimidine or pyridazine rings which are (a) fused to a 6-membered aromatic (unsaturated) heterocycle having one nitrogen atom; (b) fused to a 5- or 6-membered aromatic (unsaturated) heterocycle having two nitrogen atoms; (c) fused to a 5-membered aromatic (unsaturated) heterocycle having one nitrogen atom and one oxygen or one sulfur atom; or (d) fused to a 5-membered aromatic (unsaturated) heterocycle having one heteroatom selected from O, N or S. Heteroaryls within this definition include, but are not limited to: benzimidazolyl, benzofuranyl, benzofurazanyl, benzpyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carbolinyl, cinnolinyl, furanyl, dihydroindolyl, indolyl, indolizinyl, indazolyl, isobenzofuranyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, oxazoline, isoxazoline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridopyridinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrazolyl, tetrazolopyridinyl, thiadiazolyl, thiazolyl, thiophenyl, triazolyl, azetidinyl, aziridinyl, 1,4-dioxanyl, hexahydroazepinyl, dihydrobenzimidazolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl, dihydrofuranyl, dihydroimidazolyl, dihydroindolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl, dihydrooxazolyl, dihydropyrazinyl, dihydropyrazolyl, dihydropyridyl, dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl, dihydrotetrazolyl, dihydrothiadiazole, dihydrothiazolyl, dihydrothiophenyl, dihydrotriazolyl, dihydroazetidinyl, methylenedioxybenzyl, tetrahydrofuranyl, tetrahydrothiophenyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, benzotriazolyl, benzoxazolyl, isoxazolyl, isothiazolyl, furanyl, thiophenyl, benzothiophenyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, tetrahydroquinolinyl. In the case where the heteroaryl substituent is bicyclic and one ring is non-aromatic or does not contain heteroatoms, it is understood that the attachment is made respectively through the aromatic ring or through the ring containing heteroatoms. If the heteroaryl contains a nitrogen atom, it is understood that its corresponding N-oxide is also covered by this definition.

[0520] As used herein, "monocyclic" includes any stable polycyclic carbocyclic ring of up to 10 atoms, and can be unsubstituted or substituted. Examples of such non-aromatic monocyclic elements include, but are not limited to: cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. Examples of such aromatic monocyclic elements include, but are not limited to: phenyl. As used herein, "heteromonocyclic" includes any monocyclic ring containing at least one heteroatom.

[0521] As used herein, "bicyclic" includes any stable polycyclic carbocyclic ring of up to 10 atoms fused to a polycyclic carbocyclic ring of up to 10 atoms, where each ring is independently unsubstituted or substituted. Examples of such non-aromatic bicyclic elements include, but are not limited to: decahydronaphthalene. Examples of such aromatic bicyclic elements include, but are not limited to: naphthalene. As used herein, "heterobicyclic" includes any bicyclic ring containing at least one heteroatom.

[0522] The term "phenyl" is intended to denote an aromatic six-membered ring containing six carbons and any of its substituted derivatives.

[0523] The term "benzyl" is intended to denote a methylene group directly attached to a benzene ring. Benzyl is a methyl group in which the hydrogen is replaced by a phenyl group and any of its substituted derivatives.

[0524] The term "pyridine" is intended to denote a heteroaryl having a six-membered ring containing 5 carbon atoms and 1 nitrogen atom and any of its substituted derivatives.

[0525] The term "pyrazole" is intended to denote a heteroaryl having a five-membered ring containing three carbon atoms and two nitrogen atoms, and any of its substituted derivatives, where the nitrogen atoms are adjacent to each other.

[0526] The term "indole" is intended to denote a heteroaryl having a five-membered ring fused to a benzene ring, where the five-membered ring contains 1 nitrogen atom directly attached to the benzene ring.

[0527] The term "oxetane" is intended to denote a non-aromatic four-membered ring containing three carbon atoms and one oxygen atom and any of its substituted derivatives.

[0528] The compounds used in the methods of the present invention can be prepared by techniques well known in organic synthesis and techniques familiar to those of ordinary skill in the art. However, these may not be the only means of synthesizing or obtaining the desired compounds.

[0529] The compounds of the present invention can be prepared by the techniques described in the following literature: Vogel’s Textbook of Practical Organic Chemistry, A.I. Vogel, A.R. Tatchell, B.S. Furnis, A.J. Hannaford, P.W.G. Smith, (Prentice Hall) 5th Edition (1996), March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, Michael B. Smith, Jerry March, (Wiley-Interscience) 5th Edition (2007), and the references therein, which are incorporated herein by reference. However, these may not be the only means of synthesizing or obtaining the desired compounds.

[0530] The compounds of the present invention can be prepared by the techniques described herein or the techniques described in PCT International Publication Nos. WO / 2014 / 152013, WO / 2015 / 168286, WO / 2014 / 151959, WO / 2014 / 152018, and WO / 2014 / 151936, the respective contents of which are incorporated herein by reference.

[0531] The various R groups attached to the aromatic rings of the compounds disclosed herein can be added to the rings by standard procedures such as those shown in the following literature: Advanced Organic Chemistry: Part B: Reaction and Synthesis, Francis Carey and Richard Sundberg, (Springer) 5th Edition (2007), the content of which is incorporated herein by reference.

[0532] Another aspect of the present invention includes the compounds of the present invention as pharmaceutical compositions.

[0533] As used herein, the term "pharmaceutically active agent" refers to any substance or compound suitable for administration to a subject and that provides a biological activity or other direct effect in the treatment, cure, mitigation, diagnosis, or prevention of a disease, or that affects the structure or any function of the subject. Pharmaceutically active agents include, but are not limited to, substances and compounds described in the following references: Physicians’ Desk Reference (PDR Network, LLC; 64th edition; November 15, 2009) and “Approved Drug Products with Therapeutic Equivalence Evaluations” (U.S. Department Of Health And Human Services, 30th edition, 2010), which are incorporated herein by reference. Pharmaceutically active agents having pendant carboxylic acid groups can be modified according to the invention using standard esterification reactions and methods readily available and known to those of ordinary skill in the art of chemical synthesis. In the case where the pharmaceutically active agent does not have a carboxylic acid group, one of ordinary skill in the art will be able to design and incorporate a carboxylic acid group into the pharmaceutically active agent, whereupon esterification can then be carried out, provided that the modification does not interfere with the biological activity or effect of the pharmaceutically active agent.

[0534] The compounds of the present invention may be in the form of salts. As used herein, "salt" refers to a salt of a compound of the present invention, which is modified by preparing an acid or base salt of the compound. In the case of a compound for treating a disease, the salt is pharmaceutically acceptable. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral acid or organic acid salts of basic residues (such as amines); alkali metal salts or organic salts of acidic residues (such as phenols). The salts can be made with organic or inorganic acids. Such acid salts are chlorides, bromides, sulfates, nitrates, phosphates, sulfonates, formates, tartrates, maleates, malates, citrates, benzoates, salicylates, ascorbates, etc. Phenol salts are alkaline earth metal salts, sodium salts, potassium salts or lithium salts. In this regard, the term "pharmaceutically acceptable salt" refers to relatively non-toxic inorganic and organic acid or base addition salts of the compounds of the present invention. These salts can be prepared in situ during the final isolation and purification of the compounds of the present invention, or by reacting the purified compounds of the present invention in their free base or free acid form with a suitable organic or inorganic acid or base, respectively, and isolating the salts so formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthoate, mesylate, gluconate, lactobionate and laurylsulfonate, etc. (see, for example, Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0535] Salts or pharmaceutically acceptable salts are contemplated for all compounds disclosed herein. In some embodiments, pharmaceutically acceptable salts or salts of any of the above compounds of the present invention.

[0536] As used herein, "treatment" refers to preventing, slowing down, halting or reversing the progression of a disease or infection. Treatment may also mean improving one or more symptoms of a disease or infection. One embodiment of "treating gout" is delaying or preventing the onset, progression or reducing the severity of gout.

[0537] As used herein, "normalizing", in terms of normalizing the concentration in a diseased subject, means increasing or decreasing the concentration such that the concentration is closer to the concentration in a subject without the disease.

[0538] As used herein, "elevating", such as the concentration of RBP4 being elevated in a diseased subject, means that the concentration of RBP4 is increased compared to the concentration in a subject without the disease.

[0539] The compounds of the present invention can be administered in various forms, including those detailed herein. The treatment with the said compounds can be a component of combination therapy or adjuvant therapy, i.e., treating a subject in need of the drug by combining one or more compounds of the present invention, or administering additional drugs for the said disease thereto. The combination therapy can be sequential therapy, where the patient is treated first with one drug and then with another, or the two drugs are administered simultaneously. Depending on the dosage form employed, they can be administered independently by the same or two or more different routes of administration.

[0540] As used herein, "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent or vehicle for delivering the compounds of the present invention to an animal or a human. The vehicle can be liquid or solid and is selected according to the intended mode of administration under consideration. Liposomes are also pharmaceutically acceptable carriers.

[0541] The dosage of the compounds administered in the treatment will vary according to a variety of factors, such as the pharmacodynamic properties of the particular chemotherapeutic agent and its mode and route of administration; the age, sex, metabolic rate, absorption efficiency, health status and body weight of the recipient; the nature and degree of the symptoms; the type of concurrent therapy administered; the frequency of treatment; and the desired therapeutic effect.

[0542] The dosage unit of the compounds for the methods of the present invention can contain a single compound or a mixture thereof with other agents. The compounds can be administered in oral dosage forms such as tablets, capsules, pills, powders, granules, elixirs, tinctures, suspensions, syrups and emulsions. The compounds can also be administered intravenously (bolus or infusion), intraperitoneally, subcutaneously or intramuscularly, or directly into or onto the site of infection, e.g., by injection, topical application or other methods, all using dosage forms well known to those of ordinary skill in the art of pharmacy.

[0543] The compounds used in the method of the present invention can be administered in admixture with a suitable pharmaceutical diluent, extender, excipient or carrier (collectively referred to herein as pharmaceutically acceptable carriers), which are appropriately selected according to the intended form of administration and in accordance with conventional pharmaceutical practice. The unit will be in a form suitable for oral, rectal, topical, intravenous or direct injection or parenteral administration. The compounds can be administered alone or in admixture with a pharmaceutically acceptable carrier. The carrier can be solid or liquid, and the type of carrier is generally selected based on the type of administration used. The active agent can be co-administered in the form of tablets or capsules, liposomes, in the form of agglomerated powders or liquids. Examples of suitable solid carriers include lactose, sucrose, gelatin and agar. Capsules or tablets are easy to formulate and are made easy to swallow or chew; other solid forms include granules and bulk powders. Tablets may contain suitable binders, lubricants, diluents, disintegrants, colorants, flavorants, flow-inducing agents and melting agents. Examples of suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents (including esters), emulsions, syrups or elixirs, suspensions, solutions and / or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifiers, suspending agents, diluents, sweeteners, thickening agents and melting agents. Oral dosage forms optionally contain flavorants and colorants. Parenteral and intravenous forms may also include minerals and other materials to make them compatible with the type of injection or delivery system selected.

[0544] Techniques and compositions for preparing dosage forms useful in the present invention are described in the following references: 7 Modern Pharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors, 1979); Pharmaceutical Dosage Forms: Tablets (Lieberman et al., 1981); Ansel, Introduction to Pharmaceutical Dosage Forms, Second Edition (1976); Remington's Pharmaceutical Sciences, 17th Edition (Mack Publishing Company, Easton, Pa., 1985); Advances in Pharmaceutical Sciences (David Ganderton, Trevor Jones, Eds., 1992); Advances in Pharmaceutical Sciences Vol. 7. (David Ganderton, Trevor Jones, James McGinity, Eds., 1995); Aqueous Polymeric Coatings for Pharmaceutical Dosage Forms (Drugs and the Pharmaceutical Sciences, Series 36 (James McGinity, Ed., 1989); Pharmaceutical Particulate Carriers: Therapeutic Applications: Drugs and the Pharmaceutical Sciences, Vol 61 (Alain Rolland, Ed., 1993); Drug Delivery to the Gastrointestinal Tract (Ellis Horwood Books in the Biological Sciences. Series in Pharmaceutical Technology; J.G. Hardy, S.S. Davis, Clive G. Wilson, Eds.); Modem Pharmaceutics Drugs and the Pharmaceutical Sciences, Vol 40 (Gilbert S. Banker, Christopher T. Rhodes, Eds.).All of the above publications are hereby incorporated by reference.

[0545] Tablets may contain suitable binders, lubricants, disintegrants, coloring agents, flavoring agents, flow inducing agents, and melting agents. For example, for oral administration in the form of tablet or capsule dosage units, the active pharmaceutical ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as lactose, gelatin, agar, starch, sucrose, glucose, methylcellulose, magnesium stearate, dibasic calcium phosphate, calcium sulfate, mannitol, sorbitol, and the like. Suitable binders include starch, gelatin, natural sugars such as glucose or β-lactose, corn sweeteners, natural and synthetic gums such as gum arabic, tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, waxes, and the like. Lubricants used in these dosage forms include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agar, bentonite, xanthan gum, and the like.

[0546] The compounds used in the methods of the present invention may also be administered in the form of a liposome delivery system such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids such as cholesterol, stearylamine, or phosphatidylcholine. The compounds may be administered as a component of a tissue-targeted emulsion.

[0547] The compounds used in the methods of the present invention may also be conjugated to a soluble polymer as a targetable drug carrier or as a prodrug. Such polymers include polyvinylpyrrolidone, pyran copolymers, poly(hydroxypropylmethacrylamide)-phenol, poly(hydroxyethyl aspartamide)-resorcinol, or poly(ethylene oxide)-polylysine substituted with palmitoyl residues. In addition, the compounds may be conjugated to a class of biodegradable polymers useful for achieving controlled release of drugs such as polylactic acid, polyglycolic acid, copolymers of polylactic acid and polyglycolic acid, poly(ε-caprolactone), polyhydroxybutyrate, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates, and crosslinked or amphiphilic block copolymers of hydrogels.

[0548] Gelatin capsules may contain the active ingredient compound and a powdered carrier such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, and the like. Similar diluents can be used to make compressed tablets. Both tablets and capsules can be made into immediate release products or sustained release products to provide continuous release of the drug over several hours. Compressed tablets can be coated with sugar coating or film coating to mask any unpleasant taste and protect the tablets from atmospheric effects, or coated with enteric coating to selectively disintegrate in the gastrointestinal tract.

[0549] For oral administration in liquid dosage forms, the oral pharmaceutical components are combined with any oral, non-toxic, pharmaceutically acceptable inert carriers such as ethanol, glycerol, water, etc. Examples of suitable liquid dosage forms include solutions or suspensions in water, pharmaceutically acceptable fats and oils, alcohols or other organic solvents (including esters), emulsions, syrups or elixirs, suspensions, solutions and / or suspensions reconstituted from non-effervescent granules, and effervescent preparations reconstituted from effervescent granules. Such liquid dosage forms may contain, for example, suitable solvents, preservatives, emulsifying agents, suspending agents, diluents, sweetening agents, thickening agents, and melting agents.

[0550] Oral liquid dosage forms may contain coloring and flavoring agents to increase patient acceptance. Generally, water, suitable oils, saline, aqueous dextrose (glucose) solutions, and related sugar solutions and diols (such as propylene glycol or polyethylene glycol) are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain water-soluble salts of the active ingredient, suitable stabilizers, and, if necessary, buffering substances. Antioxidants such as sodium bisulfite, sodium sulfite, or ascorbic acid, alone or in combination, are suitable stabilizers. Citric acid and its salts and sodium edetate are also used. In addition, parenteral solutions may contain preservatives such as benzalkonium chloride, methylparaben, propylparaben, and chlorobutanol. Suitable pharmaceutical carriers are described in the standard reference book in the field: Remington's Pharmaceutical Sciences, Mack Publishing Company.

[0551] The compounds used in the method of the present invention may also be administered intranasally using a suitable intranasal vehicle, or by the transdermal route, using transdermal skin patches in forms known to those of ordinary skill in the art. For administration in the form of a transdermal delivery system, the administration is generally continuous rather than intermittent throughout the dosing regimen.

[0552] Parenteral and intravenous forms may also include minerals and other materials to make them compatible with the type of injection or delivery system selected.

[0553] Each embodiment disclosed herein is considered applicable to each other disclosed embodiment. Accordingly, all combinations of the various elements described herein are within the scope of the present invention.

[0554] The present invention will be better understood by reference to the following experimental details, but those skilled in the art will readily understand that the specific experiments described in detail are merely illustrative of the present invention as more fully described in the claims that follow.

[0555] Experimental Details

[0556] Example 1:

[0557] AsFigure 1 As shown, single oral and intravenous administration of compound 1 induced a significant decrease in circulating levels of serum RBP4 in wild-type mice.

[0558] The main objectives of this study were to determine the targeting of compound 1 in mice and to determine whether compound 1 was active in mice. To perform this assay, the effects of oral and intravenous administration of compound 1 on the dynamics of plasma RBP4 levels in male CD-1 mice were investigated. Aliquots of the collected plasma samples were used to analyze plasma RBP4 concentrations using a standard ELISA protocol (Adipogen RBP4 (Mouse / Rat) Dual ELISA Kit) as described by the manufacturer of the ELISA kit. Plasma RBP4 concentrations were measured at 10 time points after IV or PO compound administration. Due to the small amount of blood in mice, blood samples can be collected from a single animal up to 3 times. Samples from different animals were used to analyze RBP4 concentrations at different time points. Pre-dose plasma samples were collected from all animals participating in the study, and the average pre-dose RBP4 concentration was used as a baseline. The experimental design of this study is shown in Table 1.

[0559] Table 1: Experimental design and mouse grouping

[0560]

[0561] a Blood was collected from 3 untreated mice as baseline controls.

[0562] b The intravenous route is through the tail vein.

[0563] For IV administration, compounds were prepared as solutions in 3% DMA / 45% PEG300 / 12% ethanol / 40% sterile water. For PO administration, compounds were prepared as 2% in 0.9% saline. In summary, oral (5 mg / kg) and intravenous (2 mg / kg) administration of compound 1 resulted in a significant reduction in plasma RBP4 concentrations in male CD-1 mice. The maximum mean plasma RBP4 reduction was 84.9% 12 hours after oral (PO) administration and 81.4% 8 hours after intravenous (IV) administration. The findings provide evidence for the in vivo activity of the test compound in mice.

[0564] Embodiment 2:

[0565] Emerging evidence suggests that in addition to its retinol transport function, RBP4 can play a pathogenic role in several common diseases such as insulin resistance, type 2 diabetes (T2D), metabolic syndrome, and low-grade vascular inflammation. There is substantial evidence supporting the association between elevated circulating levels of RBP4 and the development of NAFLD.

[0566] Lee 2016 recently reported a transgenic mouse model ("adi-hRBP4 mice") in which human RBP4 can be specifically expressed in adipocytes. Due to the increased production of human RBP4 in mouse adipose tissue, the circulating levels of RBP4 in these transgenic mice are mildly elevated. These transgenic mice gain weight and develop hepatic steatosis. In fact, when fed a regular diet, these mice develop NAFLD at 3 - 4 months of age; when fed a high-fat diet (HFD), the deterioration of their metabolic phenotype is faster than that of their littermate controls. Thus, adi-hRBP4 mice are a transgenic model of hepatic steatosis.

[0567] Compound 1 (an advanced RBP4 antagonist) was evaluated in adi-hRBP4 mice. In this experiment, male adi-hRBP4 mice were divided into three age-matched treatment groups. The first group was fed a regular diet (hereinafter referred to as the "regular diet group"), the second group was fed a high-fat diet (HFD) (hereinafter referred to as the "HFD-only group"), and the third group was fed HFD and compound 1 (hereinafter referred to as the "treatment group"). Compound 1 was formulated into food to provide a daily oral dose of 20 mg compound 1 / kg mouse body weight to the treatment group. The treatment period was four (4) weeks. The following variables were measured: body weight, food consumption, liver lipids, blood chemistry, urine analysis, liver histology, kidney histology, retina - histology, and ERG.

[0568] The experimental design is outlined in Figure 2 and the experimental data of this experiment are shown in Figures 3 - 14

[0569] Both human and mouse RBP4 are present in the mouse circulatory system (serum), as Figure 4 shown. Human RBP4 is expressed and secreted in adipose tissue, while mouse RBP4 is mainly produced in the liver. However, the circulating concentration of human RBP4 produced in adipose tissue in untreated animals is approximately 5% of that of mouse RBP4 mainly produced in the liver. As Figure 4 shown, in HFD-fed adi-hRBP4 mice, the administration of compound 1 significantly reduced the circulating levels of both human and mouse serum RBP4 levels.

[0570] As Figure 6 and Figure 7 ​As shown, it was also found that Compound 1 significantly reduced HFD-induced weight gain, although there was no difference in food intake between the HFD-only group and the treatment group ( Figure 8 ). Compared with the HFD-only group, it was also found that Compound 1 significantly reduced the concentrations of liver triglycerides and hepatic free fatty acids ( Figure 9 and 10 ).

[0571] Figure 14 It was shown that fatty acids can serve as ligands for RBP4, indicating that RBP4 may be involved in the transport of fatty acids from adipose tissue to the liver. Compound 1 and other RBP4 antagonists can inhibit this transport because they compete for binding to the same ligand-binding pocket in RBP4.

[0572] Therefore, reducing the levels of RBP4 in humans and mice is associated with the significant efficacy of Compound 1 in reducing weight gain and normalizing the concentrations of triglycerides and free fatty acids in the liver.

[0573] This data indicates that Compound 1 and similar compounds can be used to treat diseases within the scope of human NAFLD. Compound 1 and similar compounds are particularly suitable for treating diseases within the scope of NAFLD in patients with elevated serum RBP4 levels.

[0574] Example 3: Treatment of gout

[0575] Hepatic steatosis in NAFLD induces upregulation of xanthine oxidase in the liver, leading to increased uric acid production. In people with NAFLD, elevated circulating uric acid concentrations can cause gout symptoms. NAFLD is closely associated with the development of hyperuricemia.

[0576] Administering Compound 1 to the transgenic mice described in Example 2 significantly reduced the circulating levels of uric acid. Therefore, Compound 1 and similar compounds can be used to treat gout and gouty arthritis in patients with NAFLD. Compound 1 and similar compounds can also be used to treat gout and gouty arthritis in patients without NAFLD.

[0577] Example 4: Combination treatment of gout

[0578] Since Compound 1 inhibits the production of uric acid in the liver, Compound 1 and similar compounds can be used in combination with drugs that induce the excretion of uric acid in the kidneys. An example of such a drug is DPOFA, which is exemplified in PCT International Publication No. WO / 2017 / 083652, which is incorporated herein by reference. In view of the disclosure herein, these two compounds have complementary mechanisms of action. Specifically, DPOFA increases the excretion of uric acid through urine, while Compound 1 inhibits the upregulation of NAFLD-induced xanthine oxidase (XO) in the liver. XO is the enzyme responsible for uric acid synthesis in the liver, which is the main site of uric acid production. Thus, in the combination, Compound 1 reduces the production of uric acid, while DPOFA increases the uric acid excreted from the body, thereby treating the subject.

[0579] Example 5: Efficacy in a Mammalian Model

[0580] The effectiveness of the compounds listed herein was tested in adi-hRBP4 mice. The adi-hRBP4 mouse model exhibits a moderate increase in human RBP4 expression in mouse adipose tissue and is considered a model of hepatic steatosis. The compounds were administered orally at 20 mg / kg for 4 weeks. The serum levels of human and mouse RBP4 were reduced in the treated animals. In the treated mice, the levels of RBP4 and free fatty acids (FFAs) in the liver were reduced. (1) The levels of free fatty acids in the serum, (2) triglycerides in the liver, and (3) serum uric acid were also reduced.

[0581] Example 6: Administration to a Subject

[0582] A subject with NAFLD is administered an amount of Compound 1. The amount of the compound is effective to treat the subject.

[0583] A patient with NAFLD and elevated circulating levels of serum RBP4 is administered an amount of Compound 1. The amount of the compound is effective to treat the subject.

[0584] A subject with hepatic steatosis is administered an amount of Compound 1. The amount of the compound is effective to treat the subject.

[0585] A subject with non-alcoholic steatohepatitis (NASH) is administered an amount of Compound 1. The amount of the compound is effective to treat the subject.

[0586] A subject with cirrhosis is administered an amount of Compound 1. The amount of the compound is effective to treat the subject.

[0587] A subject with hepatocellular carcinoma is administered an amount of Compound 1. The amount of the compound is effective to treat the subject.

[0588] Administer a certain amount of Compound 1 to a subject with gout. The amount of the compound can effectively treat the subject.

[0589] The structures of Compounds 2 - 4 are shown below.

[0590]

[0591] Administer a certain amount of Compound 2 to a subject with NAFLD. The amount of the compound can effectively treat the subject.

[0592] Administer a certain amount of Compound 2 to a subject with hepatic steatosis. The amount of the compound can effectively treat the subject.

[0593] Administer a certain amount of Compound 2 to a subject with non - alcoholic steatohepatitis (NASH). The amount of the compound can effectively treat the subject.

[0594] Administer a certain amount of Compound 2 to a subject with liver cirrhosis. The amount of the compound can effectively treat the subject.

[0595] Administer a certain amount of Compound 2 to a subject with hepatocellular carcinoma. The amount of the compound can effectively treat the subject.

[0596] Administer a certain amount of Compound 2 to a subject with gout. The amount of the compound can effectively treat the subject.

[0597] Administer a certain amount of Compound 3 to a subject with NAFLD. The amount of the compound can effectively treat the subject.

[0598] Administer a certain amount of Compound 3 to a subject with hepatic steatosis. The amount of the compound can effectively treat the subject.

[0599] Administer a certain amount of Compound 3 to a subject with non - alcoholic steatohepatitis (NASH). The amount of the compound can effectively treat the subject.

[0600] Administer a certain amount of Compound 3 to a subject with liver cirrhosis. The amount of the compound can effectively treat the subject.

[0601] Administer a certain amount of Compound 3 to a subject with hepatocellular carcinoma. The amount of the compound can effectively treat the subject.

[0602] Administer a certain amount of Compound 3 to a subject with gout. The amount of the compound can effectively treat the subject.

[0603] Administer an amount of Compound 4 to a subject with NAFLD. The amount of the compound can effectively treat the subject.

[0604] Administer an amount of Compound 4 to a subject with hepatic steatosis. The amount of the compound can effectively treat the subject.

[0605] Administer an amount of Compound 4 to a subject with non-alcoholic steatohepatitis (NASH). The amount of the compound can effectively treat the subject.

[0606] Administer an amount of Compound 4 to a subject with liver cirrhosis. The amount of the compound can effectively treat the subject.

[0607] Administer an amount of Compound 4 to a subject with hepatocellular carcinoma. The amount of the compound can effectively treat the subject.

[0608] Administer an amount of Compound 4 to a subject with gout. The amount of the compound can effectively treat the subject.

[0609] Example 7: DPOFA in combination with Compound 1

[0610] Administer an amount of Compound DPOFA in combination with Compound 1 to a subject with gout. The amounts of DPOFA and Compound 1 can effectively treat the subject with gout. The combination is more effective in treating gout than each compound administered alone.

[0611] Example 8: DPOFA in combination with Compound 1

[0612] Administer an amount of Compound DPOFA in combination with Compound 1 to a subject with gout and NAFLD. The amounts of DPOFA and Compound 1 can effectively treat the subject with gout and NAFLD. The combination is more effective in treating gout and NAFLD than each compound administered alone.

[0613] Example 9: DPOFA in combination with Compound 1 prevents gout in a subject

[0614] Administer an amount of Compound DPOFA in combination with Compound 1 to a subject with a history of gout attacks or at risk of gout attacks. The amounts of DPOFA and Compound 1 can effectively prevent gout attacks in the subject. The combination is more effective in preventing gout than each compound administered alone.

[0615] Example 10: In vivo activity: PK properties of Compound 1 and Compound 3 in rodents.

[0616] In naive male CD-1 mice and adult Sprague-Dawley male rats, both Compound 1 and Compound 3 had favorable PK profiles ( Figure 15 ). The compounds showed moderate to low clearance and good half-lives (t 1 / 2 ) in both species. Adequate exposures (AUC last ) and good oral bioavailability (%F) were also obtained. The good oral bioavailability, moderate half-lives, appropriate exposures, and good ADME properties obtained in mice fully justified the use of Compound 1 and Compound 3 to establish PK / PD and ultimately proof-of-concept in a mouse animal model.

[0617] Example 11: In Vivo Activity: PK / PD Correlation of Compound 1 in Mice.

[0618] Prior to testing Compound 1 in a transgenic model of hepatic steatosis in mice, an acute dosing study of Compound 1 in mice was conducted to measure the effect of the compound on circulating plasma RBP4 levels and to establish a PK / PD correlation ( Figure 16 ). After a single oral dose (5 mg / kg) of Compound 1, a maximum reduction of 85% in plasma RBP4 was observed ( Figure 16 A), while after a 2 mg / kg intravenous dose, plasma RBP4 was reduced by 81% ( Figure 16 B). The reduction of serum RBP4 in vivo after oral and intravenous administration of Compound 1 ( Figure 16 A, B) showed a good correlation with the compound concentration in plasma ( Figure 16 C, D). The long-term exposure and moderate to low clearance of Compound 1 obtained after a single oral dose were closely related to the extent of RBP4 reduction (85% reduction at the 12-hour time point) and the duration of the RBP4 reduction effect (71% reduction at the 24-hour time point).

[0619] Furthermore, the magnitude of RBP4 reduction was closely related to the predicted free drug concentration of Compound 1 in plasma, which exceeded the concentration required to disrupt the RBP4-TTR interaction measured in an in vitro HTRF assay. These data confirmed the in vivo targeting of Compound 1 and revealed a good PK / PD relationship between compound exposure and the biological response in mice, and further confirmed the properties of Compound 1 in a transgenic model of hepatic steatosis in mice.

[0620] Example 12: In a transgenic model of hepatic steatosis in mice, Compound 1 reduced the circulating levels of adipose-derived RBP4.

[0621] As previously reported by Blaner and colleagues (Lee, S.-A. et al. 2016), a genetic model of hepatic steatosis in mice can be generated by targeting the mouse Rosa26 locus with a human RBP4 (hRBP) cDNA construct with a loxP-neor-stop cassette. To specifically express hRBP4 in mouse adipocytes, knock-in mice were bred with adiponectin-Cre mice. Specific expression of human RBP4 in mouse adipocytes did not result in a significant increase in the circulating levels of RBP4, and there were no changes in retinol levels in plasma, liver, and adipose tissue, while causing obesity, impaired glucose tolerance, and a significant increase in liver triglyceride (TG) levels (Lee, S.-A. et al. 2016).

[0622] To examine the effect of compound 1 on the metabolic parameters of male adi-hRBP4 mice, the compound was administered as a formulation at a dose of 20 mg / kg per day to HFD food for 29 days. When the animals were switched from standard food to a high-fat diet (60% of calories from fat), the administration of the compound started at 20 weeks of age. Long-term oral administration of compound 1 caused a 90% reduction in the circulating levels of serum RBP4 in mice and humans (secreted from adipose tissue)( Figure 17 ). In untreated adi-hRBP4 mice, the circulating levels of adipose-derived human RBP4 (2 - 3 μg / mL) accounted for a small fraction of 3 - 5% of mouse-specific serum RBP4 produced mainly in the liver( Figure 17 ). Notably, this modest increase in the circulating levels of RBP4 conferred by adipocyte secretion was sufficient to trigger the induction of a strong metabolic phenotype.

[0623] Example 13: Compound 1 reduces weight gain in obese adi-hRBP4 mice.

[0624] During the 29-day study period, the weight gain of adihRBP4 mice on a high-fat diet was significantly higher than that of transgenic animals maintained on standard food( Figure 18 A). Five days after the start of high-fat feeding, there was a significant statistical difference in the percentage of weight gain between mice fed food and HFD adi-hRBP mice( Figure 18 A). By administering compound 1, the weight gain of HFD animals was significantly reduced. Nineteen days after high-fat diet feeding, there was a significant statistical difference in weight gain between compound 1-treated and untreated HFD mice( Figure 18 A). At the end of the 29-day treatment (2.2 ± 1.7 g), it was 53% less than that of untreated HFD animals (4.7 ± 1.6 g). The reduction in weight gain of compound 1-treated adi-RBP4 mice was not related to a decrease in food intake, as compound 1 did not alter the consumption of HFD foodFigure 10 B).

[0625] Example 14: Compound 1 reduces hepatic lipid levels in obese adi-hRBP4 mice.

[0626] According to previous reports (Lee, S.-A. et al. 2016), the levels of hepatic free fatty acids (FFAs) and triglycerides (TGs) in adi-hRBP4 mice maintained on a high-fat diet were significantly higher than those in transgenic animals maintained on standard chow ( Figure 19 ; for both FFAs and TGs, P < 0.0001). Administration of Compound 1 decreased the FFA level by 30% ( Figure 11 A; P = 0.0107), and the TG level by 29% ( Figure 11 B; P = 0.0104).

[0627] Consistent with the dynamics of hepatic TG accumulation, histological examination of frozen liver sections stained with Oil Red O ( Figure 20 A) and hepatic steatosis grading ( Figure 20 A) confirmed that adi-hRBP4 mice maintained on an HFD had significantly more steatosis compared to chow-fed transgenic mice that did not show evidence of hepatic steatosis.

[0628] Significant improvement in hepatic steatosis was observed in adi-hRBP4 obese mice treated with Compound 1, with lipid droplets showing a decreasing trend compared to untreated adi-hRBP4 mice maintained on an HFD ( Figure 18 B). Hepatic steatosis grading ( Figure 12 B) showed that the degree of steatosis was significantly reduced by 43% (P < 0.001) in HFD-fed adi-hRBP4 mice treated with Compound 1, further confirming the ability of Compound 1 to alleviate hepatic steatosis in adi-hRBP4 mice.

[0629] Example 15: Binding of fatty acid ligands to RBP4.

[0630] It is not clear how a modest increase in RBP4 expression in adipose tissue promotes the development of hepatic steatosis. It is speculated that this ability of RBP4 may be related to the binding and transport of endogenous non-retinoid ligands, the nature and quantity of which are not yet clear at this point. Consistent with the ability of RBP4 to interact with non-retinoid ligands, previous crystallographic studies of heterologously expressed RBP4 have shown that accidentally discovered fatty acid ligands (oleic acid and linoleic acid) from the expression host bind in the ligand-binding pocket of RBP4 (Huang, H.-J. et al. 2009; Nanao, M.S. et al. 2009).

[0631] The latest X-ray crystallography and mass spectrometry findings reported by Monaco and colleagues confirmed that RBP4 can bind fatty acids (Perduca, M. et al. 2018). The ability of retinol-binding proteins to bind hydrophobic non-retinoid ligands may be common, as illustrated by the recent description of the interaction of cellular retinol-binding protein 1 with certain cannabinoids (Silvaroli, J.A. et al. 2019).

[0632] The affinities of palmitic acid, oleic acid, linoleic acid, and docosahexaenoic acid for RBP4 were tested using a SPA assay that measures the displacement of radioactive retinol in purified human RBP4 ( Figure 21 A). These competitive binding experiments confirmed that the tested fatty acids function as weak RBP4 ligands. Our docking model is also consistent with the ability of RBP4 to bind fatty acid ligands. Figure 13 B shows the overlap of compound 3 and the fatty acids palmitic acid, oleic acid, linoleic acid, and docosahexaenoic acid docked in our 3FMZ model, as well as all-trans retinol docked within our 5NU7 (holo-RBP4) model. The figure shows a good overlap between all-trans retinol and compound 3, as both present similar geometries and project their polar fragments towards the opening of the binding cavity, where they make H-bond interactions with Gln98 and Arg121, respectively. Similar to all-trans retinol and compound 3, the fatty acids also extend their hydrophobic tails through the narrow β-barrel and into the β-ionone pocket, and their polar carboxylic acids bind to residues closer to the opening of the binding cavity, namely Leu36 and Phe36.

[0633] Discussion

[0634] On the one hand, the present invention relates to small molecules for the treatment of NAFLD diseases. The specific use of small molecules as non-retinoid RBP4 antagonists for the treatment of non-alcoholic fatty liver disease (NAFLD) diseases is disclosed herein. The compounds listed herein have been shown to bind RBP4 in vitro at biologically significant concentrations and / or to antagonize the RBP4-TTR interaction in vitro. The additional compounds described herein are analogs of the compounds listed herein that similarly bind RBP4 in vitro at biologically significant concentrations and antagonize the RBP4-TTR interaction in vitro.

[0635] Fenritinide [N-(4-hydroxy-phenyl)retinamide, 4HRP] is a retinoid drug previously developed for the treatment of cancer. Similar to other retinoid drugs, fenritinide is non-specific and thus capable of binding to multiple drug targets. Its anti-cancer activity is attributed to the ability to generate reactive oxygen species and induce apoptosis in malignant tumor cells. Some other activities of this compound are mediated through the action of this compound as a ligand for the nuclear receptor RAR. The ability of fenritinide to act as an RBP4 ligand has been well characterized within the different activity profiles of fenritinide. For example, fenritinide has been found to bind to RBP4, displace all-trans retinol of RBP4 (Berni 1992), disrupt the RBP4-TTR interaction (Berni 1992, Schaffer 1993), reduce serum RBP4 and retinol (Adams 1995), and inhibit intraocular all-trans retinol uptake and slow down the visual cycle (Radu 2005). However, it is known from the literature that the beneficial effects of fenritinide in diabetes and obesity models are not mediated by its activity as an RBP4 antagonist. Koh 2012 reported that fenritinide was able to improve insulin sensitivity and reduce hepatic lipid levels in obese mice. This observation is consistent with earlier reports which showed that long-term administration of fenritinide could prevent high-fat diet-induced obesity, insulin resistance, and hepatic steatosis (Preitner 2009). However, it was convincingly shown in Preitner that, given that fenritinide was equally effective in reducing fat mass in RBP4 knockout animals, its activity as an RBP4 antagonist did not mediate the beneficial effects of fenritinide on reducing HFD-induced obesity. Motani 2009 also demonstrated that the ability of fenritinide to improve insulin sensitivity in HFD-fed mice was independent of its activity as an RBP4 antagonist. Overall, although Koh (and the earlier work of Preitner and Motani) showed the beneficial effects of fenritinide in relevant HFD-induced obesity models, the earlier work of Preitner and Motani convincingly attributed these beneficial effects to activities of fenritinide that were independent of its involvement with RBP4. For these reasons, in view of Koh 2012 and Preitner 2009, one of ordinary skill in the art would not reasonably expect the administration of an RBP4 antagonist to treat NAFLD disease.

[0636] Fenritinide also has toxicity and teratogenicity. Therefore, the safety profile of fenritinide may not be compatible with long-term administration in individuals with non-life-threatening conditions.

[0637] As briefly discussed above, Koh 2012 discloses the use of fenritinide (“FEN”) in genetically obese (ob / ob) mice. In Koh, the ob / ob mice were divided into two groups and both groups were fed a high-fat diet (HFD), but FEN was administered to only one group. Koh 2012 reported that FEN reduced weight gain, insulin resistance, and fatty liver in HFD-fed ob / ob mice. Koh also reported that FEN decreased circulating RBP4 and TTR levels (page 371). In addition, Koh reported a decrease in RBP4 levels in the adipose tissue of FEN-treated obese mice (right column, page 374). Koh noted that “after fenritinide treatment, lower levels of circulating lipids may be transported to tissues including the liver” (right column, page 374). In contrast to Koh 2012, Lee 2016 showed that in mice not fed a high-fat diet, free fatty acids were redistributed from adipose tissue to the liver rather than to all tissues as implied by Koh.

[0638] Koh 2012 further showed that plasma adiponectin levels differed between vehicle-treated and FEN-treated mice, which may be related to reduced triglyceride (TG) synthesis and increased free fatty acid (FFA) oxidation in FEN-treated mice. However, due to an increase in the level of the adiponectin-stimulated transcription factor PPARα, Koh later noted that their findings suggested that FEN may generally affect fatty acid oxidases in obese mice (pages 374 to 375). Koh further concluded that lower hepatic fat accumulation resulted from increased fatty acid oxidation rather than reduced lipogenesis (page 375). In contrast, Lee 2016 found that increased TNF and leptin expression led to RBP4-induced inflammation in adipose tissue, resulting in increased lipolysis. Lee 2016 further pointed out that the free fatty acids generated by lipolysis were taken up by the liver, which led to increased TG levels and hepatic steatosis.

[0639] Thus, it is shown not only that the positive properties of fenritinide in the HFD-induced obesity model are independent of its activity as an RBP4 antagonist, but also that Koh 2012 proposed an incorrect mechanism for its proposed treatment with fenritinide.

[0640] In addition, Koh 2012 did not disclose the use of any RBP4 antagonist to treat non-alcoholic fatty liver disease (NAFLD), and the mice used in Koh's study were not a specific model of NAFLD. In contrast, the data in the present application are from a transgenic model of hepatic steatosis, a NAFLD disease.

[0641] Therefore, Koh 2012 does not provide any reasonable insights into the successful treatment of NAFLD with RBP4 antagonists.

[0642] Although Lee 2016 discloses substantial evidence correlating elevated circulating levels of RBP4 with the development of NAFLD (page 1534), Lee 2016 also points out that some studies have reported evidence of a lack of association between RBP4 and NAFLD (page 1543). Thus, the full role of elevated circulating RBP4 levels in the development of NAFLD remains unclear.

[0643] Lee 2016 further states that the development of obesity leads to increased expression of RBP4 in adipocytes (page 1534). However, Lee specifically states that "whether RBP4 can truly stimulate hepatic steatosis in a liver-autonomous manner remains to be determined." Similar to Koh 2012, Lee 2016 found no increase in the rate of de novo lipogenesis in the livers of adi-hRBP4 mice (page 1540). However, unlike Koh, Lee did not find any statistically significant genotype-dependent differences in free fatty acid (FFA) oxidation. Instead, Lee states that his data support the conclusion that increased hepatic uptake of circulating fatty acids (FFAs) "substantially" contributes to the fatty liver phenotype observed in adi-hRBP4 mice.

[0644] However, Lee 2016 does not show what effect an RBP4 antagonist has on adi-hRBP4 mice or on any subject. For example, none of the experiments disclosed in Lee 2016 administered any RBP4 antagonist to a subject. In fact, Lee 2016 does not suggest that RBP4 antagonists can be used to treat NAFLD.

[0645] In the present application, the applicant has in particular shown that RBP4 antagonists can normalize the concentration of triglycerides and free fatty acids in the liver, thereby treating NAFLD disease.

[0646] Currently, there is no FDA-approved pharmaceutical therapy for any form of NAFLD. The present invention identifies non-retinoid RBP4 antagonists that can be used to treat NAFLD and other conditions characterized by the excessive accumulation of RBP4 in adipocytes. Without wishing to be bound by any scientific theory, since the accumulation of RBP4 in adipocytes appears to be the direct cause of fatty liver in NAFLD, the compounds described herein are disease modifiers because they directly address the cause of NAFLD. The present invention provides novel methods of treatment that will treat patients with NAFLD as well as patients with conditions characterized by excessive RBP4 in adipocytes.

[0647] References

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[0654] Koh IU, Jun HS, Choi JS, Lim JH, Kim WH, Yoon JB, et al. Fenretinide ameliorates insulin resistance and fatty liver in obese mice. Biol Pharm Bull 2012; 35:369–375. Lee, Seung-Ah et al. “Adipocyte-Specific Over-Expression of Retinol Binding Protein 4 (RBP4) Causes Hepatic Steatosis in Mice.” Hepatology (Baltimore, Md.) 64.5 (2016):1534–1546. PMC.

[0655] Lee, S.A., Yuen, J.J., Jiang, H., Kahn, B.B. & Blaner, W.S. Adipocyte-specific overexpression of retinol-binding protein 4 causes hepatic steatosis in mice. Hepatology 64, 1534–1546 (2016).

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Claims

1. A method for treating a subject with non-alcoholic fatty liver disease (NAFLD), the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound is a non-retinoid retinol-binding protein 4 (RBP4) antagonist.

2. A method for treating a subject with gout, the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound is a retinol-binding protein 4 (RBP4) antagonist.

3. A method for treating a subject with non-alcoholic fatty liver disease (NAFLD) or gout, the method comprising administering to the subject a pharmaceutical composition comprising an amount of a compound effective to treat the subject, thereby treating the subject, wherein the compound has the following structure wherein L is a linking group having the following structure: and Z is a group having the following structure: wherein: R1, R2, R3, R4, and R5 are each independently H, halogen, CF3, or a C1-C4 alkyl, aryl, or heteroaryl; R6 is H, OH, halogen, or absent; ψ is absent or present, and when present is a bond; B is a substituted or unsubstituted heterobicyclic, pyridazine, pyrazole, pyrazine, thiadiazole, or triazole, wherein the heterobicyclic is not a chloro-substituted indole; and the pyrazole, when substituted, is not substituted with trifluoromethyl; B’ is a substituted or unsubstituted phenyl, pyridine, pyrimidine, benzyl, pyrrolidine, sulfolane, oxetane, CO2H, or (C1-C4 alkyl)-CO2H, wherein the substituted phenyl is not substituted with trifluoromethyl or 3-(methyl carboxylate), the substituted pyridine is not substituted with trifluoromethyl, and the substituted pyrrolidine is not substituted with hydroxamic acid, and the substituted or unsubstituted pyrrolidine is bonded to the carbonyl group through a carbon-carbon bond; A is non-existent or existent, and when existent, is B1 is a substituted or unsubstituted monocyclic, bicyclic, heteromonocyclic, heterobicyclic, benzyl, CO2H, or (C1-C4 alkyl)-CO2H, where when B1 is CO2H, then A is present and is R7 is an alkyl; X is N or CR8, where R8 is H, OH, or halogen; B2 has the following structure: wherein: α and β are each a bond that is present or absent; X1 is N, NH or NR 99 , wherein R 99 is an alkyl, alkenyl or alkynyl group; X2 is C or N; X3 is CH or N; R9, R 10 and R 11 each independently is H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl--NH2, alkyl-OAc alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3, wherein: X1, X2, and X3 are each N, α is present and β is absent; or X1 is NH, X2 is C, X3 is CH, α is absent and β is present; or X1 is N, X2 is N, X3 is CH, α is present and β is absent; or X1 is NH or NR 99 , X2 is C, X3 is N, α is absent and β is present, where when X1 is NH, X2 is C, X3 is N, α is absent and β is present, then one of R9, R 10 and R 11 is not H Alternatively, B2 has the following structure: wherein: R 12 、R 13 and R 14 each independently is H, halogen, alkyl, alkenyl, alkynyl, alkyl-OH, alkyl-NH2, alkyl-Oac, alkyl-O(CO)-alkyl, alkyl-O-alkyl, haloalkyl, cycloalkyl, O-alkyl, NH-alkyl, C(O)OH, C(O)-NH2, C(O)-N(CH3)2, C(O)-NHCH3, NHC(O)-N(CH3)2, CN or CF3, or a pharmaceutically acceptable salt thereof.

4. The method according to claim 3, wherein R1, R2, R3, R4, and R5 are each independently H, halogen, CF3, or a C1-C4 alkyl.

5. The method according to any one of claims 1-4, wherein the subject has an NAFLD disease selected from the group consisting of: hepatic steatosis (fatty liver), non-alcoholic steatohepatitis (NASH), cirrhosis, and hepatocellular carcinoma.

6. The method according to any one of claims 1-5, wherein the method further comprises the step of determining or having determined the level of RBP4 in the adipose tissue of the subject, and administering the pharmaceutical composition if the level of RBP4 in the adipose tissue is elevated.

7. The method according to any one of claims 1-6, wherein the method further comprises the step of determining or having determined the level of RBP4 in the serum of the subject, and administering the pharmaceutical composition if the level of RBP4 in the serum is elevated.

8. The method according to any one of claims 1-7, wherein the amount of the compound is effective to reduce the level of RBP4 in the adipose tissue of the subject or reduce the level of RBP4 in the serum of the subject.

9. The method according to any one of claims 1-8, wherein the amount of the compound is effective to reduce the level of uric acid in the serum of the subject.

10. The method according to any one of claims 1-9, wherein the amount of the compound is effective to normalize the concentration of triglycerides in the liver of the subject, or normalize the concentration of free fatty acids in the serum of the subject, and normalize the concentration of free fatty acids in the liver of the subject.

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