Use of 5-methoxy-2-aminoindane ("MEAI") in method for treating metabolic syndrome

Abstract

The present disclosure relates to methods of treating a metabolic condition by administering to a subject in need thereof a therapeutically effective amount of 5-methoxy-2-aminoindane or a salt thereof. The present disclosure also relates to methods of treating a metabolic condition by administering to a subject in need thereof a therapeutically effective amount of 5-methoxy-2-aminoindane, or a pharmaceutically acceptable salt thereof, and N-acylethanolamine, or a pharmaceutically acceptable salt thereof.

Description

[0001] Cross - Reference to Related Applications

[0002] This application claims priority to U.S. Provisional Application No. 63 / 408,683, filed on September 21, 2022, the content of which is hereby incorporated by reference. Technical Field

[0003] The present disclosure particularly relates to methods of treating metabolic conditions and related syndromes by administering a therapeutically effective amount of 5-methoxy-2-aminoindane (“MEAI”). In certain embodiments, the treatment methods include co-administering MEAI with one or more N-acylethanolamines, such as palmitoylethanolamide (“PEA”). Background Art

[0004] Obesity is a chronic disease that has reached epidemic proportions, with more than one-third (34.9% or 78.6 million) of U.S. adults considered obese. Obesity has been described as a catalyst for many conditions, most notably cardiovascular disease, type 2 diabetes mellitus (T2DM), and non-alcoholic fatty liver disease (NAFLD). Although several metabolic factors are associated with the development of obesity, the exact molecular mechanisms involved have not been fully understood.

[0005] Furthermore, despite the severity of obesity and its comorbidities, there are still few anti-obesity agents available on the market. The causes of obesity are attributed to dietary behavior or fast food, personality problems, depression, or genetics. Food addiction is one of the emerging hypotheses for the current epidemic spread of obesity, which is generally associated with both substance use disorders and eating disorders (most prominently binge eating). There is evidence that binge eating sugar-dense palatable foods increases extracellular dopamine in the striatum and thus has addictive potential. In addition, there seem to be several biological and psychological similarities between food addiction and drug dependence, including craving and loss of control.

[0006] However, as shown by the limited options for treating obesity and other metabolic disorders, there is still a need for improved treatment options. Summary of the Invention

[0007] In some embodiments, provided herein are methods for treating a metabolic condition, such as a metabolic condition associated with one or more metabolic syndromes, the methods comprising administering to a subject in need thereof a therapeutically acceptable amount of a pharmaceutical composition comprising 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof, thereby treating the metabolic condition. In certain embodiments, 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered in a dose of about 20 to about 520 mg. In other embodiments, 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered in a dose of about 0.5 to about 40 mg.

[0008] In some embodiments, 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered in a dose of: about 20 to about 100 mg, about 25 to about 90 mg, about 30 to about 80 mg, about 40 to about 70 mg, or about 50 to about 60 mg. In other embodiments, the dose is administered as a single dose or as more than one divided dose. In other embodiments, the dose is administered daily as a single dose or as more than one divided dose. In certain embodiments, 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered twice daily.

[0009] In some embodiments, a therapeutically effective amount of 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof comprises about 0.0084 to about 0.67 mg / kg body weight / day, about 0.33 to about 8.67 mg / kg body weight / day, about 0.33 to about 1.67 mg / kg body weight / day, about 0.42 to about 1.5 mg / kg body weight / day, about 0.5 to about 1.33 mg / kg body weight / day, about 0.67 to about 1.17 mg / kg body weight / day, or about 0.83 to about 1.0 mg / kg body weight / day.

[0010] In some embodiments, the administered pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier and / or excipient. In certain embodiments, the pharmaceutical composition is a free-flowing powder, tablet, capsule, lozenge, liquid, liquid concentrate, suspension, or syrup. In some embodiments, the pharmaceutical composition is a unit dosage form composition. In some embodiments, the amount of 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof in the unit dosage form is about 20 to about 520 mg, about 0.5 to about 40 mg, about 20 to about 100 mg, about 25 to about 90 mg, about 30 to about 80 mg, about 40 to about 70 mg, or about 50 to about 60 mg. In certain embodiments, the amount of 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is about 50 mg.

[0011] In some embodiments, the administration of the pharmaceutical composition is oral, sublingual, buccal, vaginal, rectal, parenteral, transdermal, or by inhalation. In certain embodiments, parenteral administration is intravenous, intramuscular, or subcutaneous.

[0012] In some embodiments, treating a metabolic condition, such as a metabolic condition associated with one or more metabolic syndromes, includes administering a therapeutically acceptable amount of a pharmaceutical composition comprising 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof, and administering an N-acylethanolamine or a pharmaceutically acceptable salt thereof, either alone, concurrently, or simultaneously, to treat the metabolic disorder. The N-acylethanolamine or a pharmaceutically acceptable salt thereof can be in the same or a separate pharmaceutical composition as the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof.

[0013] In some embodiments, the N-acylethanolamine or a pharmaceutically acceptable salt thereof is administered in a dose of about 200 to about 1800 mg. In some embodiments, the N-acylethanolamine or a pharmaceutically acceptable salt thereof is administered in a dose of: about 250 to about 1550, about 300 to about 1200 mg, about 350 to about 950 mg, about 400 to about 700 mg, about 450 to about 600 mg, or about 500 to about 550 mg. In other embodiments, the dose is administered as a single dose or as more than one divided dose. In other embodiments, the dose is administered daily as a single dose or as more than one divided dose. In certain embodiments, the N-acylethanolamine or a pharmaceutically acceptable salt thereof is administered twice daily.

[0014] In some embodiments, 5-methoxy-2-aminoindane and the N-acylethanolamine can be administered in a molar ratio in the range of about 1:0.2 to about 1:2000.

[0015] In some embodiments, the N-acylethanolamine is selected from N-palmitoylethanolamide (PEA), Me-palmitoylethanolamide (Me-PEA), palmitoylcyclohexylamide, palmitoylbutyramide, palmitoylisopropylamide, oleoylethanolamine (OEA), palmitoylisopropylamide (PIA), salts thereof, and any combination thereof. The various possibilities represent separate embodiments of the present disclosure. In certain embodiments, the N-acylethanolamine is PEA or a salt thereof. In certain embodiments, the N-acylethanolamine consists of PEA or a salt thereof. In certain embodiments, the N-acylethanolamine consists of PEA.

[0016] In some embodiments, a therapeutically effective amount of an N-acylethanolamine or a pharmaceutically acceptable salt thereof includes from about 2.5 to about 36.0 mg / kg body weight / day, from about 3.12 to about 31.0 mg / kg body weight / day, from about 3.75 to about 24.0 mg / kg body weight / day, from about 4.38 to about 19.0 mg / kg body weight / day, from about 5.0 to about 14.0 mg / kg body weight / day, from about 5.62 to about 12.0 mg / kg body weight / day, or from about 6.25 to about 11.0 mg / kg body weight / day.

[0017] In some embodiments, the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier and / or excipient. In certain embodiments, the pharmaceutical composition is a free-flowing powder, tablet, capsule, lozenge, liquid, liquid concentrate, suspension, or syrup. In some embodiments, the pharmaceutical composition is a unit dosage form composition.

[0018] In some embodiments, administration of the pharmaceutical composition is oral, sublingual, buccal, vaginal, rectal, parenteral, transdermal, or by inhalation. In certain embodiments, parenteral administration is intravenous, intramuscular, or subcutaneous.

[0019] In some embodiments, administration is by oral, mucosal, nasal, sublingual, inhalation, topical, rectal, vaginal, or parenteral routes. In certain embodiments, parenteral administration is intravenous, intramuscular, or subcutaneous.

[0020] In some embodiments, treating a metabolic condition includes alleviating one or more metabolic syndromes, such as lowering blood pressure, lowering blood glucose, reducing body fat around the waist, normalizing abnormal cholesterol or triglyceride levels, alleviating obesity, reducing overweight, reducing body weight, increasing lean mass, reducing fat mass, alleviating adiposity, increasing energy expenditure, improving glycemic control, reducing hepatic steatosis, reducing glucose uptake, reducing food intake, maintaining glucose homeostasis, reducing dyslipidemia, or preserving liver function. In certain embodiments, improving glycemic control involves improving glucose metabolism, lowering fasting blood glucose levels, or lowering insulin levels. In some embodiments, increasing energy expenditure involves increasing oxygen consumption and carbon dioxide emission, increasing fat oxidation, or increasing locomotive activity.

[0021] In some embodiments, the improved metabolic syndrome involves alleviating obesity. In some embodiments, treating the metabolic syndrome involves reducing overweight associated with obesity. In some embodiments, treating the metabolic syndrome preserves the lean mass of the subject. In some embodiments, treating the metabolic syndrome reduces the fat mass of the subject. In some embodiments, treating the metabolic syndrome alleviates adiposity of the subject.

[0022] In some embodiments, the improved metabolic syndrome includes increased energy expenditure. In some embodiments, treating the metabolic syndrome increases energy expenditure and food consumption remains unchanged. In some embodiments, treating the metabolic syndrome increases energy expenditure and increases fat utilization. In some embodiments, treating the metabolic syndrome increases energy expenditure and normalizes spontaneous activity without overstimulation.

[0023] In some embodiments, the improved metabolic syndrome includes improved glycemic control. In some embodiments, treating the metabolic syndrome reverses hyperglycemia, glucose intolerance, or hyperinsulinemia. In some embodiments, treating the metabolic syndrome treats hepatic steatosis. In some embodiments, treating hepatic steatosis involves reducing one or more of hepatic lipid accumulation, hepatic triglyceride levels, or hepatic cholesterol levels. In some embodiments, treating the metabolic syndrome maintains glucose homeostasis. In some embodiments, maintaining glucose homeostasis involves improving one or more of glucose tolerance, attenuating insulin resistance, reducing dyslipidemia, or reducing hepatic lipid accumulation.

[0024] In some embodiments, a pharmaceutical composition comprising 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is used to treat the metabolic syndrome according to any of the foregoing embodiments.

[0025] In some embodiments, a pharmaceutical composition comprising 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof and palmitoylethanolamide or a pharmaceutically acceptable salt thereof is used to treat the metabolic syndrome according to any of the foregoing embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The foregoing summary, as well as the following detailed description of the disclosure, will be better understood when read in conjunction with the accompanying drawings. For purposes of illustrating the disclosure, the drawings show some, but not all, alternative embodiments. It should be understood, however, that the disclosure is not limited to the precise arrangements and instrumentalities shown. These figures, which are incorporated in and constitute a part of this specification, help to explain the principles of the disclosure.

[0027] Figure 1 shows the acute effects of MEAI administration on food intake patterns and energy utilization. The experimental design is shown ( Figure 1A ), cumulative food intake and total food intake ( Figure 1B , C), cumulative water intake and total water intake ( Figure 1D , E), respiratory exchange ratio (RER) ( Figure 1F ), oxygen consumption rate ( Figure 1G ), carbon dioxide emission rate ( Figure 1H ), total energy expenditure (TEE) ( Figure 1I , J), fat oxidation ( Figure 1K) and carbohydrate oxidation( Figure 1L ). Data are expressed as mean ± SEM from 6 - 8 mice per group. *P < 0.05 versus vehicle - treated group.

[0028] Figure 2 shows acute alterations in activity profiles after MEAI administration. Total locomotor activity( Figure 2A ), ambulatory movement( Figure 2B ), ambulatory speed( Figure 2C ), wheel running distance( Figure 2D ), and total pedometer count( Figure 2E ) are shown. Data are expressed as mean ± SEM from 6 - 8 mice per group. *P < 0.05 versus vehicle - treated group.

[0029] Figure 3 Shows the acute effect of MEAI on sweet preference. Percentage of sucrose preference compared to sterile water during a 48 - hour test period is shown. Data are expressed as mean ± SEM from 8 mice per group. *P < 0.05 versus vehicle - treated group.

[0030] Figure 4 shows that long - term MEAI administration attenuates weight gain and body composition changes associated with obesity. The experimental design for testing the efficacy of MEAI in an HFD - induced obesity model( Figure 4A ), time - course changes in body weight( Figure 4B ), total body weight at the end of the experiment( Figure 4C ), change in total body weight at the end of the experiment( Figure 4D ), percentage of lean mass in total body weight( Figure 4E ), lean mass in grams( Figure 4F ), percentage of fat mass in total body weight( Figure 4G ), and fat mass in grams( Figure 4H ) are shown. Data are expressed as mean ± SEM from 8 - 11 mice per group. *P < 0.05 versus STD vehicle; #P < 0.05 versus HFD vehicle.

[0031] Figure 5 shows the effect of long - term MEAI administration on food consumption and energy metabolism. Rate of food consumption per hour( Figure 5A ), cumulative food consumption, total food consumption in g / day, total food consumption in kcal / day( Figure 5B , C, D), cumulative water intake( Figure 5E ), respiratory exchange ratio (RER)( Figure 5F ), oxygen consumption rate( Figure 5G ), carbon dioxide emission rate( Figure 5H ), total energy expenditure rate( Figure 5I ), J), fat oxidation(Figure 5K ) and carbohydrate oxidation ( Figure 5L ). Data are presented as mean ± SEM from 8 - 11 mice per group. *P < 0.05 versus STD vehicle; #P < 0.05 versus HFD vehicle.

[0032] Figure 6 shows the spontaneous activity after long - term MEAI administration in HFD - induced obesity. Shown are the 24 - hour time - course changes in ambulatory activity ( Figure 6A ), locomotor movement ( Figure 6B ), locomotor movement speed ( Figure 6C ), total distance traveled ( Figure 6D ), running wheel movement distance ( Figure 6E ), wheel speed ( Figure 6F ), and a graph showing the percentage of time that mice spent engaged in various activities ( Figure 6G ). Data are presented as mean ± SEM from 8 - 11 mice per group. *P < 0.05 versus STD vehicle; #P < 0.05 versus HFD vehicle.

[0033] Figure 7 shows the effects of long - term MEAI administration on glucose tolerance and insulin sensitivity. Shown are the blood glucose levels in the glucose tolerance test ( Figure 7A ), the area under the curve (AUC) values of the glucose tolerance test ( Figure 7B ), the percentage of blood glucose in the insulin tolerance test ( Figure 7C ), the AUC values of the insulin tolerance test ( Figure 7D ), fasting blood glucose levels ( Figure 7E ), serum insulin levels ( Figure 7F ), homeostasis model assessment insulin resistance (HOMA - IR) values ( Figure 7G ) and insulin sensitivity index (ISI) values ( Figure 7H ). Data are presented as mean ± SEM from 8 - 11 mice per group. *P < 0.05 versus STD vehicle; #P < 0.05 versus HFD vehicle.

[0034] Figure 8 shows the circulating lipid profiles after MEAI treatment. Shown are HDL levels ( Figure 8A ), LDL levels ( Figure 8B ), the ratio of HDL to LDL ( Figure 8C ), cholesterol levels ( Figure 8D ) and triglyceride levels ( Figure 8E ). Data are presented as mean ± SEM from 8 - 11 mice per group. *P < 0.05 versus STD vehicle; #P < 0.05 versus HFD vehicle.

[0035] Figure 9 shows the effects of MEAI on kidney weight and function. Kidney weight ( Figure 9A ), the ratio of kidney weight to body weight ( Figure 9B ), and BUN as measured by a COBAS chemistry analyzer ( Figure 9C ) are shown. Data are expressed as the mean ± SEM from 8 - 11 mice per group. *P < 0.05 versus STD vehicle; #P < 0.05 versus HFD vehicle.

[0036] Figure 10 shows that MEAI improves obesity - related hepatic steatosis. The weight of liver samples ( Figure 10A ), the ratio of liver weight to body weight ( Figure 10B ), ALT levels ( Figure 10C ), AST levels ( Figure 10D ), ALP levels ( Figure 10E ), hepatic triglyceride content ( Figure 10F ), hepatic cholesterol content ( Figure 10G ), the percentage of Oil Red O - stained area ( Figure 10H ), and Oil Red O - stained samples demonstrating lipid vacuoles in hepatocytes ( Figure 10I ) are shown after long - term treatment with MEAI compared to vehicle. Data are expressed as the mean ± SEM from 8 - 11 mice per group. *P < 0.05 versus STD vehicle; #P < 0.05 versus HFD vehicle. DETAILED DESCRIPTION

[0037] Compounds derived from 2 - aminoindane can be used in the methods disclosed herein. Such compounds are shown to selectively bind to the dopamine D3 receptor. U.S. Patent No. 5,708,018 discloses some 2 - aminoindane derivatives and postulates that these 2 - aminoindane derivatives can be used to treat CNS disorders associated with the dopamine D3 receptor. One such compound is 5 - methoxy - 2 - aminoindane (“MEAI”), the chemical formula of which is:

[0038]

[0039] Other 2 - aminoindane derivatives that can be used in the present disclosure can be represented by compounds of formula I:

[0040]

[0041] wherein R1 and R2 are each independently selected from H, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, aryl, heteroaryl, heterocycloaliphatic, -O(C1-C8)alkyl, OH, -OSO2CF3, -OSO2-(C1-C8)alkyl, -SOR5, -CO2R5, -CONR5R6, -COR5, -CF3, CN, -SR5, -SO2NR5R6, -SO2R5, -OCO-(C1-C8)alkyl, -NCO-(C1-C8)alkyl, -CH2O-(C1-C8)alkyl, -(C1-C6)alkyl-OH, -NHSO2R5 and halogen, or alternatively, R1 and R2 together with two or more phenyl carbon atoms form a -X1-(CR5R6)m-X2- ring, wherein X1 and X2 are each independently selected from C, O, NH or S, and m is 1, 2, 3 or 4;

[0042] R3 and R4 are each independently selected from H, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl and -(CH2)p-thienyl, where p is 1, 2, 3 or 4, or alternatively, R3 and R4 are linked together to form a heterocycle (heterocycloaliphatic or heteroaryl), the heterocycle containing the nitrogen atom to which they are attached; and

[0043] R5 and R6 are each independently selected from H, (C1-C8)alkyl, (C2-C8)alkenyl, (C3-C8)cycloalkyl and aryl.

[0044] In some embodiments, the 2-aminoindane derivatives represented by Formula I as presented herein are defined as follows:

[0045] R1 and R2 are each independently selected from H, (C1-C8)alkyl, (C2-C8)alkenyl, (C2-C8)alkynyl, (C3-C8)cycloalkyl, aryl, -OCH3, OH, -OSO2CF3, -OSO2CH3, -SOR5, -CO2R5, -CONR5R6 -COR5, -CF3, -CN, -SR5, -SO2NR5R6, -SO2R5, -CH2-OH, halogen, phthalimido, phenylthio, pyrrolyl, pyrrolinyl, oxazolyl, or alternatively, R1 and R2 together with two or more phenyl carbon atoms form an -O(CH2)mO- ring, where m is 1 or 2;

[0046] R3 and R4 together with the nitrogen atom to which they are attached are linked together to form a heterocyclic ring containing 4 to 8 carbon atoms; and

[0047] R5 and R6 are each independently selected from H, (C1-C8)alkyl, (C2-C8)alkenyl and (C3-C8)cycloalkyl.

[0048] Non-limiting examples of 2-aminoindane derivatives include:

[0049] (1) 5-Methoxy-2-aminoindane;

[0050] (2) 5,6-Dimethoxy-2-aminoindane;

[0051] (3) 5-Methoxy-2-(N-propylamino)indane;

[0052] (4) 5,6-Dimethoxy-2-(N-propylamino)indane;

[0053] (5) 5,6-Dimethoxy-2-(di-N-butylamino)indane;

[0054] (6) 5-(Trifluoromethylsulfonyloxy)-6-hydroxy-2-(di-N-propylamino)indane;

[0055] (7) 5-(Trifluoromethylsulfonyloxy)-2-(N-propylamino)indane;

[0056] (8) 5,6-(Di-trifluoromethylsulfonyloxy)-2-(N-propylamino)indane;

[0057] (9) 5,6-Dimethoxy-2(pyrrolidino)indane;

[0058] (10) 5-(Trifluoromethylsulfonyloxy)-6-acetoxy-2-(di-N-propylamino)indane;

[0059] (11) 5-Trifluoromethanesulfonyloxy-6-methoxy-2-(di-N-propylamino)indane;

[0060] (12) 5,6-Ethylenedioxy-2-(di-N-propylamino)indane;

[0061] (13) 5,6-Methylenedioxy-2-(di-N-propylamino)indane;

[0062] (14) 5-Hydroxy-2-(n-propylamino)indane;

[0063] (15) 5,6-Dihydroxy-2-(n-propylamino)indane;

[0064] (16) 4-Methyl-2-aminoindane;

[0065] (17) 4,5-Di-methyl-2-aminoindane;

[0066] (18) 5,6-Di-methyl-2-aminoindane;

[0067] (19)6-Methyl-2-aminoindan;

[0068] (20)4-Fluoro-2-aminoindan;

[0069] (21)5-(Isopropyl)-2-aminoindan;

[0070] (22)4,6-Dimethyl-2-aminoindan;

[0071] (23)4,7-Dimethyl-2-aminoindan;

[0072] (24)5-(tert-Butyl)-2-aminoindan;

[0073] (25)5-Propyl-2-aminoindan;

[0074] (26)5-Fluoro-2-(di-N-propylamino)indan;

[0075] (27)6-Methylenedioxy-2-(di-N-propylamino)indan;

[0076] (28)5,6-Dimethoxy-2(pyrrolidino)indan;

[0077] (29)5,6-(Di-methoxycarbonyl)-2-(di-N-propylamino)indan;

[0078] (30)5-(Methoxycarbonyl)-6-hydroxy-2-(di-N-propylamino)indan;

[0079] (31)5-Bromo-2-(dipropylamino)indan;

[0080] (32)(6-Methylthio-indan-2-yl)-dipropyl-amine;

[0081] (33)(6-Methylsulfonyl-indan-2-yl)-dipropyl-amine;

[0082] (34)(6-Methylsulfinyl-indan-2-yl)-dipropyl-amine;

[0083] (35)2-Dipropylamino-indan-5-carbaldehyde;

[0084] (36)(5-Iodo-indan-2-yl)-dipropyl-amine;

[0085] (37)(4-Iodo-indan-2-yl)-dipropyl-amine;

[0086] (38)2-Dipropylamino-indan-5-yl tosylate;

[0087] (39) 2-Dipropylamino-6-hydroxy-indan-5-yl 4-methylbenzenesulfonate;

[0088] (40) N-[2-(Benzyl-propylamino)-indan-5-yl]-4-methylbenzene-sulfonamide;

[0089] (41) N-[2-(Benzyl-propyl-amino)-indan-5-yl]methanesulfonamide;

[0090] (42) 2-[2-(Benzyl-propyl-amino)-indan-5-yl]-isoindole-1,3-dione;

[0091] (43) Benzyl-propyl-(6-pyrrol-1-yl-indan-2-yl)-amine;

[0092] (44) Propyl-(6-pyrrol-1-yl-indan-2-yl)-amine;

[0093] (45) Propyl-(6-pyrrolidin-1-yl-indan-2-yl)-amine;

[0094] (46) Dipropyl-(6-pyrrolidin-1-yl-indan-2-yl)-amine;

[0095] (47) Cyclopropanecarboxylic acid-[2-(benzyl-propyl-amino)-indan-5-yl]acetamide;

[0096] (48) N-[2-(Benzyl-propyl-amino)-indan-5-yl]propanamide;

[0097] (49) N-[2-(Benzyl-propyl-amino)-indan-5-yl]-2,2-dimethylpropanamide;

[0098] (50) 5-(2-Propenyloxy)-2-(di-N-propylamino)-indan;

[0099] (51) 5,6-Di-toluenesulfonyloxy-2-(di-N-propylamino)indan;

[0100] (52) 5-Methanesulfonyloxy-2-(di-N-propylamino)indan;

[0101] (53) 5-Methoxycarbonyl-2-(di-N-propylamino)indan;

[0102] (54) 5-Carboxamido-2-(di-N-propylamino)indan;

[0103] (55) 5,6-Di-trifluoromethanesulfonyloxy-2-(propylamino)indan;

[0104] (56) 4-Methyl-2-(di-N-propylamino)indane;

[0105] (57) 4,5-Di-methyl-2-(di-N-propylamino)indane;

[0106] (58) 5,6-Di-methyl-2-(di-N-propylamino)indane;

[0107] (59) 5-Methyl-2-(di-N-propylamino)indane;

[0108] (60) 4-Fluoro-2-(N-propyl)aminoindane;

[0109] (61) 5-(Isopropyl)-2-(di-N-propylamino)indane;

[0110] (62) 5-(Isopropyl)-2-(N-propylamino)indane;

[0111] (63) 4,6-Dimethyl-2-(di-N-propylamino)indane;

[0112] (64) 4,7-Dimethyl-2-(di-N-propylamino)indane;

[0113] (65) 5-Propyl-2-(di-N-propylamino)indane;

[0114] (66) 5-(tert-Butyl)-2-(di-propylamino)indane (5-(t-butyl)-2-(dim-propylamino)indan);

[0115] (67) 5-Trifluoromethyl-2-(di-N-propylamino)indane;

[0116] (68) 5-Sulfoxamido-2-(di-N-propylamino)indane (5-sulfoxamido-2-(di-N-

[0117] propylamino)indane);

[0118] (69) 5-(3-Thienyl)-2-(di-N-propylamino)indane;

[0119] (70) 5-Ethynyl-2-(di-N-propylamino)indane;

[0120] (71) 5-Acetyl-2-(di-N-propylamino)indane;

[0121] (72) 5-Cyano-2-(di-N-propylamino)indane;

[0122] (73)5-Methoxycarbonyl-6-acetoxy-2-(di-N-propylamino)indane;

[0123] (74)5-Methoxycarbonyl-6-trifluoromethanesulfonyloxy-2-(di-N-propylamino)indane;

[0124] (75)5-Methoxycarbonyl-6-methoxy-2-(di-N-propylamino)indane;

[0125] (76)5-Formyl-6-methoxy-2-(di-N-propylamino)indane;

[0126] (77)5-Hydroxymethyl-6-methoxy-2-(di-N-propylamino)indane;

[0127] (78)5-Carboxy-6-methoxy-2-(di-N-propylamino)indane;

[0128] (79)5-Acetyl-6-methoxy-2-(di-N-propylamino)indane;

[0129] (80)5-Carboxamide-6-methoxy-2-(di-N-propylamino)indane;

[0130] (81)5-Ethynyl-6-methoxy-2-(di-N-propylamino)indane;

[0131] (82)5-Cyano-6-methoxy-2-(di-N-propylamino)indane; and

[0132] (83)5,6-Di-(hydroxymethyl-2-(di-N-propylamino)indane.

[0133] In some embodiments described herein, the 2-aminoindane derivative represented by Formula I is any one of the above compounds (1)-(13), wherein the phenyl moiety is substituted by one or two -OCH3 or -OSO2CF3 groups, or the phenyl moiety bears an -O(CH2)mO- ring fused thereto, where m is 1 or 2. The structural formulas of Compounds 1-13 are depicted in Table A below herein.

[0134] Table A

[0135]

[0136]

[0137]

[0138] N-acylethanolamines (NAEs) are a class of fatty acid amides, lipid-derived signaling molecules. They are formed when one of several types of acyl groups is linked to the nitrogen atom of ethanolamine. These amides can conceptually be formed from fatty acids and ethanolamine, with the release of a water molecule, but the known biosynthesis uses specific phospholipase D to cleave the phospholipid moiety from N-acylphosphatidylethanolamine. Examples of N-acylethanolamines include arachidonoylethanolamine (amide of arachidonic acid (20:4ω-6) and ethanolamine), N-palmitoylethanolamine (amide of palmitic acid (16:0) and ethanolamine), N-oleoylethanolamine (amide of oleic acid (18:1) and ethanolamine), N-stearoylethanolamine (amide of stearic acid (18:0) and ethanolamine), and N-docosahexaenoylethanolamine (amide of docosahexaenoic acid (22:6) and ethanolamine).

[0139] Palmitoylethanolamide (PEA, also known as N-(2-hydroxyethyl)hexadecanamide; hydroxyethyl palmitamide; palmitoyl hydroxyethylamide; N-palmitoylethanolamine; and palmitylethanolamide) is an example of an NAE and is an endogenous fatty acid amide belonging to the class of nuclear factor agonists. The chemical structure of PEA is: PEA has been shown to bind to receptors (nuclear receptors) in the cell nucleus and to exert various biological functions related to chronic pain and inflammation. Studies have shown that PEA interacts with different non-CB1 / CB2 receptors, suggesting that PEA utilizes a unique "parallel" endocannabinoid signaling system. Growing evidence that the production and inactivation of PEA can occur independently of the production and inactivation of AEA and 2-AG further supports this concept. Many of the biological effects of PEA on cells can be attributed to its affinity for PPARs (especially PPAR-α and PPAR-γ). PEA has been shown to have an affinity for the cannabinoid-like G-coupled receptors GPR55 and GPR119, as well as the transient receptor potential vanilloid type 1 receptor (TRPV1). PEA has been shown to have anti-inflammatory, antinociceptive, neuroprotective, and anticonvulsant properties.

[0140] In various embodiments, methods of treating metabolic conditions are disclosed herein, such as by reducing one or more metabolic syndromes generally, and more specifically but not exclusively, methods for treating obesity are disclosed, the methods comprising administering to a subject in need thereof a therapeutically effective amount of MEAI or a pharmaceutically acceptable salt thereof. In some embodiments, the method further comprises administering an N-acylethanolamine or a pharmaceutically acceptable salt thereof.

[0141] In some embodiments, the present disclosure also provides preclinical evidence of the efficacy of MEAI in regulating energy metabolism and alleviating obesity and its associated metabolic abnormalities. In some embodiments, MEAI demonstrates significant effectiveness in preventing or alleviating various conditions associated with metabolic syndrome. In addition to alleviating obesity and reducing body weight, MEAI can also maintain glucose homeostasis, reduce dyslipidemia, and preserve liver function, potentially improving fat utilization and oxidation. In some embodiments, MEAI may have the potential to serve as a novel treatment option for obesity and its associated metabolic disorders.

[0142] Definition:

[0143] "Metabolic syndrome" refers to a cluster of symptoms (which can occur together) that can increase a subject's risk of developing a metabolic condition or being associated with a metabolic condition, such as obesity, heart disease, stroke, or type 2 diabetes. These symptoms, individually or in combination, include elevated blood pressure, hyperglycemia and / or insulin resistance (type 2 diabetes), excessive body fat around the waist, and abnormal cholesterol (low HDL and / or high LDL) triglyceride levels, with or without fatty liver disease, obesity, overweight, overweigh, excessive fat mass, morbid obesity, reduced energy expenditure, abnormal blood glucose control, increased hepatic steatosis, excessive sugar intake, excessive food intake or consumption, abnormal glucose homeostasis, increased dyslipidemia, or abnormal liver function. In some cases, a person exhibiting metabolic syndrome may have excessive body fat around the waist and / or be obese.

[0144] "Metabolic condition" refers to a group of conditions associated with one or more metabolic syndromes. Examples of metabolic conditions include obesity, diabetes, diabetes associated with obesity, cardiovascular disease, non-alcoholic steatohepatitis, fatty liver disease, or dyslipidemia. Examples of dyslipidemia can include subjects with elevated cholesterol levels, elevated triglyceride levels, and / or a reduced HDL / LDL ratio.

[0145] "Overweight" and "obesity" refer to abnormal or excessive fat accumulation that poses a risk to health. "Overweight" means that a subject has a weight higher than what is considered a healthy weight for a given height and a body mass index greater than a first threshold and less than a second threshold. The second threshold can be understood to be greater than the first threshold. For human subjects, the first threshold for body mass index can be 25.0, and the second threshold for body mass index can be 30. "Obesity" means that a subject has a weight that is also higher than what is considered a healthy weight for a given height and a body mass index greater than the second threshold.

[0146] "Type 2 diabetes" means a condition in which a subject is unable to process insulin to regulate blood glucose levels. "Type 2 diabetes" can be understood to develop from and / or be associated with overweight and / or obesity. Insulin resistance can be measured by known assays (such as homeostasis model assessment (HOMA), glucose / insulin ratio, insulin sensitivity test, insulin tolerance test, hyperinsulinemic-euglycemic clamp, or any other known assay for detecting insulin resistance).

[0147] "Isomer" means a compound having the same number and kind of atoms and thus the same molecular weight, but differing in the arrangement or configuration of the atoms in space.

[0148] "Stereoisomer" or "optically active isomer" means a stable isomer having at least one chiral atom or restricted rotation giving rise to a plane of perpendicular asymmetry (such as certain biphenyls, allenes, and spiro compounds) and capable of rotating the plane of polarized light. Because of the presence of asymmetric centers and other chemical structures in the compounds of the present disclosure that can give rise to stereoisomerism, the present disclosure contemplates stereoisomers and mixtures thereof. The compounds of the present disclosure and their salts include asymmetric carbon atoms and can thus exist as single stereoisomers, racemates, and as mixtures of enantiomers and diastereomers. Typically, such compounds are prepared as mixtures of enantiomers and diastereomers, such as racemic mixtures. However, if desired, such compounds can be prepared or isolated as pure stereoisomers, i.e., as individual enantiomers or diastereomers or as stereoisomer-enriched mixtures. As discussed in more detail below, the individual stereoisomers of a compound are prepared by synthesis from an optically active starting material containing the desired chiral center or by preparing a mixture of enantiomeric products followed by separation or resolution, e.g., conversion into a mixture of diastereomers followed by separation or recrystallization, chromatographic techniques, use of a chiral resolving agent, or direct separation of enantiomers on a chiral chromatographic column. Starting compounds of specific stereochemistry are commercially available or are prepared by the methods described below and resolved by techniques well known in the art.

[0149] The biological and pharmacological activities of compounds known in the art are sensitive to the stereochemistry of the compounds. Thus, for example, enantiomers often exhibit significantly different biological activities, including differences in pharmacokinetic properties and pharmacological properties, where the pharmacokinetic properties include metabolism, protein binding, etc., and the pharmacological properties include the type of activity exhibited, the degree of activity, toxicity, etc. Accordingly, those skilled in the art will understand that an enantiomer can be more active or can exhibit beneficial effects when enriched relative to another enantiomer or when separated from another enantiomer. In addition, those skilled in the art will know how to separate, enrich, or selectively prepare the enantiomers of the compounds of the present disclosure based on the knowledge of the present disclosure and the prior art.

[0150] Thus, although the racemic form of a drug can be used, it is generally less effective than administering an equimolar amount of the enantiomerically pure drug; indeed, in some cases, one enantiomer may be pharmacologically inactive and serve only as a simple diluent. For example, although ibuprofen was previously administered as a racemate, it has been shown that only the S-isomer of ibuprofen is effective as an anti-inflammatory agent (however, in the case of ibuprofen, although the R-isomer is inactive, it is converted in vivo to the S-isomer, and thus, the rate of action of the racemic form of the drug is lower than that of the pure S-isomer). In addition, the pharmacological activities of enantiomers can have different biological activities. For example, S-penicillamine is a therapeutic agent for chronic arthritis, while R-penicillamine is toxic. Indeed, some purified enantiomers have an advantage over the racemate because it has been reported that the purified individual isomers have a more rapid percutaneous penetration rate compared to the racemic mixture. See U.S. Patent Nos. 5,114,946 and 4,818,541.

[0151] In some embodiments, the compound is a racemic mixture of (S)- and (R)-isomers. In other embodiments, provided herein are mixtures of compounds, wherein the individual compounds of the mixture are present predominantly in the (S)- or (R)-isomer configuration. For example, the mixture of compounds has the following (S)-enantiomeric excess: greater than about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5% or more. In other embodiments, the mixture of compounds has the following (S)-enantiomeric excess: greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 65% to about 99.5%, greater than about 70% to about 99.5%, greater than about 75% to about 99.5%, greater than about 80% to about 99.5%, greater than about 85% to about 99.5%, greater than about 90% to about 99.5%, greater than about 95% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 98% to greater than about 99.5%, greater than about 99% to about 99.5% or more. In other embodiments, the mixture of compounds has the following (R)-enantiomeric purity: greater than about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5% or more. In some other embodiments, the mixture of compounds has the following (R)-enantiomeric excess: greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 65% to about 99.5%, greater than about 70% to about 99.5%, greater than about 75% to about 99.5%, greater than about 80% to about 99.5%, greater than about 85% to about 99.5%, greater than about 90% to about 99.5%, greater than about 95% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 98% to greater than about 99.5%, greater than about 99% to about 99.5% or more.

[0152] Individual stereoisomers of the compounds of the present disclosure can be synthesized and prepared from commercially available starting materials that contain asymmetric or stereogenic centers, or by preparing a racemic mixture followed by resolution methods well known to those of ordinary skill in the art. Examples of such resolution methods include: (1) attaching the enantiomer mixture to a chiral auxiliary, separating the resulting mixture of diastereomers by recrystallization or chromatography, and releasing the optically pure product from the auxiliary; (2) forming salts with an optically active resolving agent; or (3) directly separating the mixture of optical enantiomers on a chiral chromatographic column. Stereoisomeric mixtures can also be resolved into their component stereoisomers by well-known methods such as chiral-phase gas chromatography, chiral-phase high-performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent. Stereoisomers can also be obtained from stereoisomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthesis methods.

[0153] Thus, if one enantiomer is more pharmacologically active, less toxic, or has a preferred distribution in vivo than the other enantiomer, it would be therapeutically more advantageous to administer the preferred enantiomer. In this way, the patient undergoing treatment will be exposed to a lower total dose of the drug and to a lower dose of the enantiomer or inhibitor of the other enantiomer that may be toxic.

[0154] As used herein, the nomenclature of compounds, including organic compounds, can be given using common names, IUPAC, IUBMB, or CAS recommendations for nomenclature. If a name is given, one of ordinary skill in the art can readily ascertain the structure of the compound by using naming conventions to systematically reconstruct the compound structure or by using commercially available software such as CHEMDRAW TM (Cambridgesoft Corporation, U.S.A.), and the structure of the compound can be easily ascertained. Chemical names are generated using PerkinElmer Professional, 17th Edition.

[0155] The compounds of the present disclosure may contain one or more chiral centers and / or double bonds and, accordingly, exist as stereoisomers such as geometric isomers, enantiomers or diastereomers. The term "stereoisomers" as used herein consists of all geometric isomers, enantiomers or diastereomers. These compounds may be named by the symbols "R" or "S", depending on the configuration of the substituents around the stereogenic carbon atom. The present disclosure encompasses various stereoisomers of these compounds and mixtures thereof. Stereoisomers include enantiomers and diastereomers. A mixture of enantiomers or diastereomers may be named "(±)" in the nomenclature, but one skilled in the art will recognize that the structure may implicitly represent chiral centers. In some embodiments, enantiomers or stereoisomers substantially free of the corresponding enantiomers may be provided.

[0156] In one aspect, the present disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of MEAI or a salt thereof and at least one N-acylethanolamine or a salt thereof.

[0157] In another aspect, the present disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of MEAI or a salt thereof and at least one N-acylethanolamine or a salt thereof, wherein the molar ratio of MEAI to N-acylethanolamine is from about 1:0.2 to about 1:2000.

[0158] As used herein, "pharmaceutical composition" refers to a formulation of the active agents described herein with other chemical components such as physiologically suitable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate the administration of a compound to an organism. As used herein, the phrase "pharmaceutically acceptable carrier" refers to a carrier, excipient or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound. Adjuvants are included under these phrases.

[0159] The term "excipient" as used herein refers to an inert substance added to a pharmaceutical composition to further facilitate the administration of the active ingredient. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and various types of starches, cellulose derivatives, gelatin, oils such as vegetable or fish oils, and polyethylene glycol.

[0160] The term "carrier" as used herein refers to a diluent, adjuvant, excipient or vehicle with which a compound is administered. Such pharmaceutical carriers may be sterile liquids, such as water and oils. Aqueous solutions such as water or saline solutions and aqueous dextrose and glycerol solutions are preferably used as carriers, especially for injectable solutions. Suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences", 18th Edition, by E.W. Martin.

[0161] As used herein, the phrase "pharmaceutically acceptable" refers to molecular entities and compositions that are physiologically tolerable and typically do not produce an allergic or similar untoward reaction when administered to an individual. Preferably, and especially when the formulation is for human use, the term "pharmaceutically acceptable" may mean approved by a regulatory agency such as the U.S. Food and Drug Agency or listed in a recognized pharmacopeia such as the U.S. Pharmacopeia for use in animals.

[0162] As used herein, the term "N-acylethanolamine" generally refers to a class of fatty acid amides, lipid-derived signaling molecules, which are formed when one of several types of acyl groups is attached to the nitrogen atom of ethanolamine. These amides can conceptually be formed from a fatty acid and ethanolamine, with the release of a water molecule, but known biosynthesis uses a specific phospholipase D to cleave the phospholipid moiety from N-acylphosphatidylethanolamine. The suffixes -amine and -amide in these names each refer to the single nitrogen atom of ethanolamine that joins the compounds together: it is called "amine" in ethanolamine because it is considered the free terminal nitrogen in this subunit, while it is called "amide" when it is considered associated with the adjacent carbonyl of the acyl subunit. In this application, the names of these compounds may be encountered as either "amide" or "amine". The term "ethanolamine" is used in a general sense and is meant to include monoethanolamine, diethanolamine, triethanolamine, and mixtures thereof.

[0163] As used herein, the term "derivative" means a compound whose core structure is the same as or closely similar to that of an N-acylethanolamine compound, but which has a chemical or physical modification, such as a different or additional side group.

[0164] As used herein, the term "salt" refers to any form of the active ingredient in which the active ingredient is assumed to be in ionic form and is coupled with a counterion (cation or anion) or is in solution. This also includes complexes of the active ingredient with other molecules and ions, especially complexes that are complexed by ionic interactions. Pharmaceutically acceptable salts are known to those of ordinary skill in the art.

[0165] In certain embodiments, the molar ratio between MEAI and N-acylethanolamine is from about 1:0.2 to about 1:1000. In certain embodiments, the molar ratio between MEAI and N-acylethanolamine is from about 1:0.2 to about 1:900, from about 1:0.2 to about 1:800, from about 1:0.2 to about 1:700, from about 1:0.2 to about 1:600, from about 1:0.2 to about 1:500, from about 1:0.2 to about 1:400, from about 1:0.2 to about 1:300, from about 1:0.2 to about 1:200, from about 1:0.2 to about 1:100, from about 1:0.2 to about 1:50, from about 1:0.2 to about 1:40, from about 1:0.2 to about 1:30, from about 1:0.2 to about 1:20, or from about 1:0.2 to about 1:10. Each of these possibilities represents a separate embodiment of the present disclosure.

[0166] In certain embodiments, the molar ratio between MEAI and N-acylethanolamine is from about 1:0.5 to about 1:2000. In certain embodiments, the molar ratio between MEAI and N-acylethanolamine is from about 1:0.5 to about 1:1000, from about 1:0.5 to about 1:900, from about 1:0.5 to about 1:800, from about 1:0.5 to about 1:700, from about 1:0.5 to about 1:600, from about 1:0.5 to about 1:500, from about 1:0.5 to about 1:400, from about 1:0.5 to about 1:300, from about 1:0.5 to about 1:200, from about 1:0.5 to about 1:100, from about 1:0.5 to about 1:50, from about 1:0.5 to about 1:40, from about 1:0.5 to about 1:30, from about 1:0.5 to about 1:20, or from about 1:0.5 to about 1:10. Each of these possibilities represents a separate embodiment of the present disclosure.

[0167] In certain embodiments, the molar ratio between MEAI and N-acylethanolamine is from about 1:1 to about 1:2000. In certain embodiments, the molar ratio between MEAI and N-acylethanolamine is from about 1:1 to about 1:1000, from about 1:1 to about 1:900, from about 1:1 to about 1:800, from about 1:1 to about 1:700, from about 1:1 to about 1:600, from about 1:1 to about 1:500, from about 1:1 to about 1:400, from about 1:1 to about 1:300, from about 1:1 to about 1:200, from about 1:1 to about 1:100, from about 1:1 to about 1:50, from about 1:1 to about 1:40, from about 1:1 to about 1:30, from about 1:1 to about 1:20, or from about 1:1 to about 1:10. Each of these possibilities represents a separate embodiment of the present disclosure.

[0168] In certain embodiments, the pharmaceutical composition comprises from about 0.5 - 10 mg of MEAI or a salt thereof. In certain embodiments, the pharmaceutical composition comprises from about 1 - 9.5 mg, from about 1.5 - 9 mg, from about 2 - 8.5 mg, from about 2.5 - 8 mg, from about 3 - 7.5 mg, from about 3.5 - 7 mg, from about 4 - 6.5 mg, from about 4.5 - 6 mg, or from about 5 - 5.5 mg of MEAI or a salt thereof. In certain embodiments, the pharmaceutical composition comprises about 0.5 mg, about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 4 mg, about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg, about 7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg, or about 10 mg of MEAI or a salt thereof. Each of the possibilities represents a separate embodiment of the invention. In certain embodiments, the pharmaceutical composition comprises less than about 0.5 mg, less than about 1 mg, less than about 1.5 mg, less than about 2 mg, less than about 2.5 mg, less than about 3 mg, less than about 3.5 mg, less than about 4 mg, less than about 4.5 mg, less than about 5 mg, less than about 5.5 mg, less than about 6 mg, less than about 6.5 mg, less than about 7 mg, less than about 7.5 mg, less than about 8 mg, less than about 8.5 mg, less than about 9 mg, less than about 9.5 mg, or about 10 mg of MEAI or a salt thereof. Each of the possibilities represents a separate embodiment of the invention. In certain embodiments, the pharmaceutical composition comprises from about 0.5 mg to about 1 mg, from about 0.5 mg to about 1.5 mg, from about 0.5 mg to about 2 mg, from about 0.5 mg to about 2.5 mg, from about 0.5 mg to about 3 mg, from about 0.5 mg to about 3.5 mg, from about 0.5 mg to about 4 mg, from about 0.5 mg to about 4.5 mg, from about 0.5 mg to about 5 mg, from about 0.5 mg to about 5.5 mg, from about 0.5 mg to about 6 mg, from about 0.5 mg to about 6.5 mg, from about 0.5 mg to about 7 mg, from about 0.5 mg to about 7.5 mg, from about 0.5 mg to about 8 mg, from about 0.5 mg to about 8.5 mg, from about 0.5 mg to about 9 mg, or from about 0.5 mg to about 9.5 mg of MEAI or a salt thereof. Each of the possibilities represents a separate embodiment of the disclosure.

[0169] In certain embodiments, the pharmaceutical composition comprises from about 200 to 1800 mg of an N-acylethanolamine or a salt thereof. In certain embodiments, the pharmaceutical composition comprises from about 250 to 1550 mg, from about 300 to 1200 mg, from about 350 to 950 mg, from about 400 to 700 mg, from about 450 to 600 mg, or from about 500 to 550 mg of an N-acylethanolamine or a salt thereof. The various possibilities represent separate embodiments of the present disclosure. In certain embodiments, the pharmaceutical composition comprises at least about 50 mg, at least about 100 mg, at least about 150 mg, at least about 200 mg, at least about 250 mg, at least about 300 mg, at least about 350 mg, at least about 400, at least about 450 mg, at least about 500 mg, at least about 550 mg, at least about 600 mg, at least about 650 mg, at least about 700 mg, at least about 750 mg, at least about 800 mg, at least about 850 mg, at least about 900 mg, at least about 950 mg, at least about 1000 mg, at least about 1050 mg, at least about 1100 mg, at least about 1150 mg, at least about 1200 mg, at least about 1250 mg, at least about 1300 mg, at least about 1350 mg, at least about 1400 mg, at least about 1450 mg, at least about 1500 mg, at least about 1550 mg, at least about 1600 mg, at least about 1650 mg, at least about 1700 mg, at least about 1750 mg, or at least about 1800 mg of an N-acylethanolamine or a salt thereof. In certain embodiments, the pharmaceutical composition comprises about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, about 1500 mg, about 1550 mg, about 1600 mg, about 1650 mg, about 1700 mg, about 1750 mg, or about 1800 mg of an N-acylethanolamine or a salt thereof. The various possibilities represent separate embodiments of the present disclosure.

[0170] In some embodiments, a pharmaceutical composition comprising MEAI further comprises a concentration of N-acylethanolamine or a pharmaceutically acceptable salt thereof sufficient to provide a patient with a dose of N-acylethanolamine or a salt thereof of from about 2.5 mg / kg / day to about 36 mg / kg / day body weight. In certain embodiments, the pharmaceutical composition comprises a concentration of N-acylethanolamine or a pharmaceutically acceptable salt thereof sufficient to provide a patient with the following doses: from about 2.5 to about 5 mg / kg / day, from about 5 mg / kg / day to about 7.5 mg / kg / day, from about 7.5 mg / kg / day to about 10 mg / kg / day, from about 10 mg / kg / day to about 12.5 mg / kg / day, from about 12.5 mg / kg / day to about 15 mg / kg / day, from about 15 mg / kg / day to about 17.5 mg / kg / day, from about 17.5 mg / kg / day to about 20 mg / kg / day, from about 20 mg / kg / day to about 22.5 mg / kg / day, from about 22.5 mg / kg / day to about 25 mg / kg / day, from about 25 mg / kg / day to about 27.5 mg / kg / day, from about 27.5 mg / kg / day to about 30 mg / kg / day, from about 30 mg / kg / day to about 32.5 mg / kg / day, from about 32.5 mg / kg / day to about 36 mg / kg body weight / day. In certain embodiments, the pharmaceutical composition comprises a concentration of N-acylethanolamine or a pharmaceutically acceptable salt thereof sufficient to provide a patient with the following doses of N-acylethanolamine or a salt thereof: about 2.5 mg / kg / day, about 5 mg / kg / day, about 7.5 mg / kg / day, about 10 mg / kg / day, about 12.5 mg / kg / day, about 15 mg / kg / day, about 17.5 mg / kg / day, about 20 mg / kg / day, about 22.5 mg / kg / day, about 25 mg / kg / day, about 27.5 mg / kg / day, about 30 mg / kg / day, about 32.5 mg / kg / day or about 36 mg / kg body weight / day. Each of the possibilities represents a separate embodiment of the present disclosure. In certain embodiments, the pharmaceutical composition comprises a concentration of N-acylethanolamine or a pharmaceutically acceptable salt thereof sufficient to provide a patient with the following doses of N-acylethanolamine or a salt thereof: about 2.5 mg / kg / day, less than about 2.5 mg / kg / day, less than about 5 mg / kg / day, less than about 7.5 mg / kg / day, less than about 10 mg / kg / day, less than about 12.5 mg / kg / day, less than about 15 mg / kg / day, less than about 17.5 mg / kg / day, less than about 20 mg / kg / day, less than about 22.5 mg / kg / day, less than about 25 mg / kg / day, less than about 27.5 mg / kg / day, less than about 30 mg / kg / day, less than about 32.5 mg / kg / day or about 36 mg / kg body weight / day. Each of the possibilities represents a separate embodiment of the present disclosure.In certain embodiments, the pharmaceutical composition comprises a concentration of an N-acylethanolamine or a pharmaceutically acceptable salt thereof that is sufficient to provide a patient with a dose of the N-acylethanolamine or its salt of from about 2.5 mg / kg / day to about 5 mg / kg / day, from about 2.5 mg / kg / day to about 7.5 mg / kg / day, from about 2.5 mg / kg to about 10 mg / kg / day, from about 2.5 mg / kg / day to about 12.5 mg / kg / day, from about 2.5 mg / kg / day to about 15 mg / kg / day, from about 2.5 mg / kg / day to about 17.5 mg / kg / day, from about 2.5 mg / kg / day to about 20 mg / kg / day, from about 2.5 mg / kg / day to about 22.5 mg / kg / day, from about 2.5 mg / kg / day to about 25 mg / kg / day, from about 2.5 mg / kg / day to about 27.5 mg / kg / day, from about 2.5 mg / kg / day to about 30 mg / kg / day, from about 2.5 mg / kg / day to about 32.5 mg / kg / day, or from about 2.5 mg / kg / day to about 36 mg / kg body weight / day. Each of the possibilities represents a separate embodiment of the present disclosure.

[0171] In certain embodiments, the N-acylethanolamine is N-palmitoylethanolamine (PEA), Me-palmitoylethanolamide (Me-PEA), palmitoylcyclohexylamide, palmitoylbutyramide, palmitoylisopropylamide, oleoylethanolamine (OEA), palmitoylisopropylamide (PIA), or a salt thereof, or any combination thereof. Each of the possibilities represents a separate embodiment of the invention. In certain embodiments, the N-acylethanolamine is PEA or a salt thereof. In certain embodiments, the N-acylethanolamine consists of PEA or a salt thereof. In certain embodiments, the N-acylethanolamine consists of PEA.

[0172] In some embodiments, administering a pharmaceutical composition comprising an N-acylethanolamine or a pharmaceutically acceptable salt thereof and administering a pharmaceutical composition comprising 5-methoxy-2-aminoindane alone, concurrently, or simultaneously can improve the therapeutic efficacy of the administration alone, concurrently, or simultaneously compared to administering only a pharmaceutical composition comprising 5-methoxy-2-aminoindane. In some embodiments, the required therapeutic dose of 5-methoxy-2-aminoindane can be reduced when administered with an N-acylethanolamine compared to administering only 5-methoxy-2-aminoindane.

[0173] In certain embodiments, the pharmaceutical composition is formulated for systemic administration. In certain embodiments, the pharmaceutical composition is formulated for oral, buccal, nasal, sublingual, inhaled, topical, rectal, vaginal, parenteral, intravenous, intramuscular, or subcutaneous administration. In certain embodiments, the pharmaceutical composition is formulated for oral, buccal, nasal, or sublingual administration. The various possibilities represent separate embodiments of the invention. In certain embodiments, the pharmaceutical composition is formulated for oral administration. In certain embodiments, the pharmaceutical composition is formulated for buccal administration. In certain embodiments, the pharmaceutical composition is formulated for nasal administration. In certain embodiments, the pharmaceutical composition is formulated for sublingual administration.

[0174] Techniques for the formulation and administration of drugs are well known in the art and can be found, for example, in "Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, Pa. The pharmaceutical compositions of the present invention can be manufactured by methods well known in the art, such as by means of conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes.

[0175] For oral administration, the pharmaceutical composition can be readily formulated by combining the active compound with pharmaceutically acceptable carriers well known in the art. Such carriers enable the pharmaceutical composition to be formulated into tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions, etc. for oral ingestion by a patient. Pharmaceutical preparations for oral use can be prepared using solid excipients, optionally grinding the resulting mixture and processing the granule mixture as desired to obtain tablets or dragee cores after adding suitable auxiliaries. Suitable excipients are, in particular, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; starch preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, and sodium carboxymethylcellulose; and / or physiologically acceptable polymers such as polyvinylpyrrolidone (PVP). If desired, disintegrating agents such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate, can be added.

[0176] The term "oral administration" refers to any method of administration in which the active agent can be administered by swallowing, chewing, sucking, or drinking an oral dosage form. Examples of solid dosage forms include conventional tablets, multilayer tablets, capsules, cachets, etc., which release little or no drug in the mouth or in the oral cavity.

[0177] Provide a suitable coating for the sugar-coated pill core. For this purpose, a concentrated sugar solution can be used, which can optionally contain gum arabic, talc, polyvinylpyrrolidone, carbapol gel, polyethylene glycol, titanium dioxide, lacquer solution and a suitable organic solvent or solvent mixture. Dyes or pigments can be added to the tablet or sugar-coated pill coating for identification or to characterize different combinations of the active compound dosage.

[0178] Pharmaceutical compositions suitable for oral use include hard or soft sealed capsules prepared from gelatin and a plasticizer such as glycerol or sorbitol. The capsules can contain a mixture of the active ingredient with a filler such as lactose, a binder such as starch, a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In the soft capsules, the active ingredient can be dissolved or suspended in a suitable liquid such as a fatty oil, liquid paraffin or liquid polyethylene glycol. Additionally, a stabilizer can be added. All formulations for oral administration should be in a dosage suitable for the selected route of administration. For buccal and sublingual administration, the composition can take the form of tablets or lozenges formulated in a conventional manner or in a viscous carrier. Alternatively, the active ingredient can be in powder form for reconstitution with a suitable vehicle such as a sterile, pyrogen-free, water-based solution prior to use.

[0179] Pharmaceutical compositions suitable for use in the context of the present invention include compositions containing an amount of the active ingredient effective to achieve the intended purpose. More specifically, a "therapeutically effective amount" means an amount of the active ingredient effective to prevent, alleviate or improve the symptoms or side effects of a disease or disorder or to prolong the survival of the treated subject. Determination of the therapeutically effective amount is well within the ability of those skilled in the art, especially in view of the detailed disclosure provided herein. More specifically, a "mixture of therapeutically effective amounts" means an amount of at least two active ingredients, where each active ingredient alone may not be a therapeutically effective amount, or where both of the active ingredients may not be therapeutically effective amounts, yet the mixture is effective to prevent, alleviate or improve the symptoms or side effects of a disease or disorder or to prolong the survival of the treated subject. As used herein, the term "mixture" refers to the non-covalent combination of two molecules.

[0180] For any formulation used in the methods of the present invention, the dosage or therapeutically effective amount can be initially estimated from in vitro, in vivo, and cell culture assays. For example, dosages can be formulated in animal models to achieve a desired concentration or titer. Such information can be used to more accurately determine useful dosages in humans. The dosages of the various compounds in the claimed combination depend on several factors, including: the method of administration, the disease to be treated, the severity of the disease, whether the disease is to be treated or prevented, and the age, weight, and health of the human to be treated. In addition, information on the pharmacogenomics of a particular patient (the effect of genotype on the pharmacokinetic, pharmacodynamic, or efficacy characteristics of a therapeutic agent) can influence the dosage used. Continuous daily dosing may not be required; the treatment regimen may require cycles during which the drug is not administered, or the therapy can be provided on an as-needed basis during periods of acute disease exacerbation. Dose escalation may or may not be required; the treatment regimen may require a reduction in the drug dosage. The toxicity and efficacy of the active ingredients described herein can be determined in vitro, in cell cultures, or in experimental animals by standard pharmaceutical procedures. Data obtained from these in vitro and cell culture assays and animal studies can be used to formulate a dosage range for humans. The dosage can vary depending on the dosage form employed and the route of administration utilized. The exact formulation, route of administration, and dosage can be selected by the individual physician in view of the patient's condition (see, e.g., Fingl, E. et al. (1975), "The Pharmacological Basis of Therapeutics," Chapter 1, p. 1). Depending on the severity and responsiveness of the condition to be treated, the administration can be by a single or multiple administrations, where the treatment continues for several days to weeks, or until a cure is effected or until a desired level of alleviation of the disease state is achieved.

[0181] In another aspect, the present disclosure further provides dosage units comprising or consisting of the pharmaceutical compositions described above.

[0182] In certain embodiments, the dosage unit comprises the pharmaceutical composition described above. In certain embodiments, the dosage unit consists of the pharmaceutical composition described above. In certain embodiments, the dosage unit is formulated as a gel, powder, or spray. In certain embodiments, the dosage unit is formulated as a gel. In certain embodiments, the dosage unit is formulated as a powder. In certain embodiments, the dosage unit is formulated as a spray.

[0183] In another aspect, the present disclosure further provides the pharmaceutical compositions or dosage units described above for use in a method of preventing or treating a condition amenable to prevention or treatment by at least one MEAI.

[0184] As used herein, the term "treatment" includes, but is not limited to, any one or more of the following: abolishing, ameliorating, inhibiting, attenuating, alleviating, blocking, suppressing, reducing, delaying, halting, relieving, preventing, or slowing the onset of one or more symptoms or side effects of a disease or condition of the present invention.

[0185] The term "acute" refers to a condition or treatment having a relatively short time course.

[0186] As used herein, the term "chronic" means that the length of time of a disease or condition or treatment of the present invention can be weeks, months, or possibly years. The intensity of the disease or condition can vary according to various circumstances, such as patient age, temperature, season, type of disease, etc.

[0187] As used herein, the term "about", in relation to a value, multiple values, or range of values defined by a minimum and a maximum value, means a value that is 10% lower and / or higher than the corresponding value, multiple values, or range of values. For example, the phrase "about 1" means "0.9 to 1.1", the phrase "about 1 or 2" means "0.9 to 1.1 or 1.8 to 2.2", and the phrase "about 1 to about 2" means "0.9 to 2.2".

[0188] As used herein, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include multiple compounds, including mixtures thereof.

[0189] Toxicity and efficacy can be determined in cell cultures or experimental animals by standard pharmaceutical procedures, such as for determining LD 50 (the lethal dose for 50% of the population) and ED 50 (the therapeutically effective dose for 50% of the population). The dose ratio between toxicity and therapeutic effect is the therapeutic index, and it can be expressed as LD 50 / ED 50 ratio. Compositions with a large therapeutic index are preferably exhibited.

[0190] Data obtained from cell culture assays or animal studies can be used to formulate a dosage range for humans. The therapeutically effective dose achieved in one animal model can be converted for use in another animal, including humans, using conversion factors known in the art (see, for example, Freireich et al., Cancer Chemother. Reports 50(4):219 - 244(1966) and the equivalent surface area dose factors in the following table).

[0191] Table 2. Equivalent surface area dose factors.

[0192]

[0193] The dosage of such compounds is preferably within the range of circulating concentrations, which range includes an ED with little or no toxicity 50 . The dosage can vary within this range, depending on the dosage form employed and the route of administration utilized. Generally, the therapeutically effective amount can vary with the age, condition and sex of the subject and the severity of the medical condition of the subject. The dosage can be determined by a physician and adjusted as needed to accommodate the observed therapeutic effect.

[0194] Those skilled in the art will recognize that both in vivo and in vitro tests using suitable, known and generally accepted cell and / or animal models predict the ability of a test compound to treat or prevent a given disorder.

[0195] Those skilled in the art further recognize that human clinical trials in healthy patients and / or patients suffering from a given disorder, including first-in-human, dose range and efficacy trials, can be completed according to methods well known in the clinical and medical arts.

[0196] Although the present disclosure has been described with reference to certain embodiments, those skilled in the art will understand that various changes can be made and equivalents can be substituted without departing from the scope of the present disclosure. In addition, many modifications can be made to adapt a particular situation or material to the teachings of the present disclosure without departing from its scope. Accordingly, the present disclosure is not intended to be limited to the particular embodiments disclosed, but the present disclosure will include all embodiments falling within the scope of the appended claims.

[0197] The following examples are presented to more fully illustrate some embodiments of the present disclosure. However, it should in no way be construed as limiting the broad scope of the present disclosure.

[0198] Example 1: Anti-obesity effect of 5-methoxy-2-aminoindane (MEAI)

[0199] Methods

[0200] Mice. The experimental protocol used was approved by the Animal Care and Use Committee of the Hebrew University, an institute accredited by AAALAC International. Male 6-week-old C57BL / 6 mice were obtained from Envigo. Animal studies were conducted in accordance with the ARRIVE guidelines, which aim to improve the transparency and reproducibility of preclinical research. The principles of replacement, refinement, or reduction were followed to minimize the number of animals used in this study. All animals were housed under specific pathogen-free (SPF) conditions, in standard plastic cages, with no more than five animals of the same sex and dose group per cage, and natural soft sawdust was provided as bedding.

[0201] Male 6-week-old C57BL / 6 mice (Envigos, Israel) were maintained on a 12-hour light / dark cycle and had free access to food. A total of 32 animals were divided into four experimental groups (N = 8 mice / group) and received a single dose of 40 mg / kg, 60 mg / kg, or 100 mg / kg of MEAI or vehicle (sterile water), which was administered via oral gavage two hours before the dark phase. The animals' drug tolerance, food and water intake, and activity and metabolic parameters were monitored 48 hours after dosing. At the end of the experiment, the animals were euthanized, and tissues (brain, liver, fat, and kidney) and blood were collected and stored frozen for future analysis. To generate diet-induced obesity, C57Bl6 / J mice were fed a high-fat diet (HFD) (60% of calories from fat, 20% from protein, and 20% from carbohydrates; Research Diet, D12492) or a standard laboratory diet (STD, 14% fat, 24% protein, 62% carbohydrates; NIH-31 rodent diet) for 18 weeks.

[0202] Effect of MEAI on obesity. Male C57BL / 6 mice were used to establish DIO by feeding them a high-fat diet (HFD; 60% kcal fat, 20% kcal protein, and 20% kcal carbohydrates; Research Diet, D12492) for 18 weeks. After this period, the mice were treated daily by gavage with vehicle (sterile water, N = 8) or MEAI (N = 11) at a dose of 40 mg / kg / day for 28 days. Age-matched control mice (N = 10) were fed a standard diet (STD; 14% kcal fat, 24% kcal protein, 62% kcal carbohydrates; NIH-31 rodent diet) daily and received vehicle. The body weight of all mice was monitored daily, and by EchoMRI-100H TM(Echo Medical Systems LLC, Houston, TX, USA) measured total body fat and lean mass. On day 29, at the end of the experimental period, mice were euthanized by cervical dislocation under anesthesia. Kidneys, brains, livers, and fat pads were removed and weighed, and samples were snap-frozen or fixed in buffered 4% formalin. Trunk blood was collected to determine biochemical parameters.

[0203] Sucrose preference test. Thirteen-week-old male C57BL / 6 mice maintained on STD and individually housed were acclimated to two water bottles in their home cages for 48 h before testing. Baseline intake was measured by weighing the bottles. On the test day (day 1), 2 h before the onset of the dark phase, fresh water and a 1.5% sucrose solution were added to the bottles, and then the mice were treated orally with MEAI (40 mg / kg, N = 8) or sterile water (N = 8). Mice were allowed to drink freely from either bottle for 24 h, after which the bottles were weighed to measure consumption. The study was repeated for an additional day (day 2), in which the bottles were switched positions (in the cage) to account for side preferences. Sucrose and water intakes over the two days were averaged, and the sucrose preference index was calculated as the average volume of sucrose solution consumed divided by the average volume of total fluid consumed (average water plus average sucrose solution).

[0204] Multiparameter metabolic assessment. The metabolic profiles and food and water intakes of mice were evaluated by using the Promethion High-Definition Behavioral Phenotyping System (Sable Instruments, Inc., Las Vegas, NV, USA). Data acquisition and instrument control were performed using MetaScreen software version 2.2.18.0, and the raw data obtained were processed using ExpeData version 1.8.4 with an analysis script that detailed all aspects of data transformation. Mice with free access to food and water were subjected to a standard 12-h light / 12-h dark cycle, which consisted of a 48-h acclimation period followed by 24 h of sampling. Respiratory gases were measured using the draw mode, negative pressure system, with a GA-3 gas analyzer (Sable Systems, Inc., Las Vegas, NV, USA). Airflow was measured and controlled at a set flow rate of 2000 mL / min with an FR-8 (Sable Systems, Inc., Las Vegas, NV, USA). Water vapor was measured continuously, and its dilution effect on O2 and CO2 was mathematically compensated. The respiratory exchange ratio (RER) was calculated as the ratio of CO2 produced (VCO2) to O2 consumed (VO2) using the following equation (1):

[0205] RER = VCO2 / VO2 (1)

[0206] Total energy expenditure (TEE) was calculated using VO2 and RER according to the following equation (2):

[0207] TEE = VO2 × (3.815 + 1.232 × RER) (2)

[0208] Fat oxidation (FO) and carbohydrate oxidation (CHO) were calculated using VO2 and VCO2 based on equations (3) and (4), respectively:

[0209] FO = 1.69 × VO2 - 1.69 × VCO2 (3)

[0210] CHO = 4.57 × VCO2 - 3.23 × VO2 (4)

[0211] Running wheel movement and spontaneous activity. Assessment of running wheel movement and spontaneous activity was performed using a Promethion high-resolution behavioral phenotyping system (Sable Instruments, Inc., Las Vegas, NV, USA). The number of running wheel revolutions was measured using a monitor that records voluntary running wheel activity, and spontaneous activity was quantified using disruptions of an infrared XYZ beam array with a beam spacing of 0.25 cm.

[0212] Glucose tolerance test (ipGTT) and insulin tolerance test (ipITT). On day 25 of the experiment, mice were fasted overnight and then received a glucose injection (1.5 g / kg intraperitoneally) on the following day (day 26). Blood glucose levels were measured using a glucometer (Bayer, Pittsburgh, PA, USA) at 0 minutes, 15 minutes, 30 minutes, 45 minutes, 60 minutes, 90 minutes, and 120 minutes after injection. Then on the following day (day 27), the mice were fasted for 6 hours and then received insulin (0.75 U / kg, intraperitoneally; Actrapid vial, Novo Nordisk A / S, Denmark). Blood glucose levels were measured at the same intervals as described above. To assess insulin resistance, the homeostasis model assessment of insulin resistance (HOMA-IR) was calculated as fasting serum insulin ([μU / mL] × fasting plasma glucose [mmol / L] / 22.5). The relative insulin sensitivity index (ISI) was calculated as 1 / (glucose × insulin) × 1000, where glucose is expressed as mg / dL and insulin is expressed as mU / L.

[0213] Blood and urine biochemistry. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), cholesterol, triglycerides, high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were determined using a Cobas C-111 chemistry analyzer (Roche, Switzerland). Blood urea nitrogen (BUN) was calculated based on the serum urea level as follows: BUN (mg / dL) = urea (mg / dL) / 2.1428. Fasting blood glucose was measured using a blood glucose meter (Bayer, Pittsburgh, PA). Serum insulin was determined using an ultrasensitive mouse insulin ELISA kit (Crystal Chem, Inc., Elk Grove Village, IL, USA).

[0214] Liver triglyceride and cholesterol content. Liver tissue was extracted as described in (Tam et al., 2012), and its cholesterol and triglyceride content was determined using a Cobas C-111 chemistry analyzer (Roche, Switzerland).

[0215] Histopathology. First, 5-μm paraffin-embedded liver sections from 5 animals per group were stained with hematoxylin-eosin. Liver images were captured using a Zeiss Axio Scope A1 optical microscope (Carl Zeiss AG, Jena, Germany) equipped with a Zeiss AxioCam ICc5 color camera. Ten random 40× fields of view were collected from each animal to obtain representative images.

[0216] Oil Red O staining. Liver cryosections (8 μm) were stained with Oil Red O (catalog number ab150678; Abcam) following the manufacturer's protocol. Images were obtained as described above. For quantitative analysis of Oil Red O staining, the area of lipid droplets in liver cryosections was measured using ImageJ software.

[0217] Statistics. Data were expressed as mean ± SEM. Statistical analysis was performed using GraphPad Prism 6.0 software (GraphPad Software, CA, USA). Unpaired two-tailed Student's t-test was used to determine differences between two groups. For comparisons involving multiple groups and time-dependent variables, ANOVA was employed, followed by Tukey's multiple comparison test. Statistical significance was considered when the p-value was less than 0.05.

[0218] Results

[0219] To evaluate the acute effects of MEAI on food intake patterns and respiratory parameters, a single dose of 40 mg / kg, 60 mg / kg, or 100 mg / kg was administered two hours before the onset of the dark phase, as Figure 1A depicted. The results showed that the drug was well tolerated, with no observable behavioral changes at the 40 mg / kg and 60 mg / kg doses. However, two subjects in the 100 mg / kg group died within hours of drug administration, indicating reduced tolerance at this dose in combination with the stress induced by the metabolic tests and, therefore, excluding them from the analysis. Minor changes in feeding patterns were observed during the active (dark) and inactive (light) phases after MEAI administration, but these changes did not reach statistical significance( Figure 1B , C). Additionally, there were no significant changes in water consumption( Figure 1D , E). In contrast, acute MEAI administration caused significant changes in respiratory parameters. The increase in RER at the 60 mg and 100 mg doses during the light phase( Figure 1F ) was attributed to higher rates of oxygen consumption (VO2) and carbon dioxide emission (VCO2) (as Figure 1G demonstrated). A significant dose-dependent increase in TEE( Figure 1I , J) and corresponding elevations in FO and CHO rates( Figure 1K , L) were evident across the 24-hour period examined, indicating changes in the energy profile.

[0220] Next, the acute effects of MEAI on the pattern of activity were investigated. In summary, the total number of beam breaks recorded, which represents a combination of walking and fine motor movements, did not change significantly( Figure 2A ). Notably, MEAI produced a significant and dose-dependent increase in directed spontaneous activity and velocity, such as moving around the cage for feeding, drinking, and grooming at doses of 40 mg / kg and above( Figure 2B , C). Although there were minor elevations in the pattern of voluntary wheel running at 40 mg / kg and 60 mg / kg, the 100 mg / kg dose group experienced a significant inhibition of wheel running( Figure 2D ). Finally, MEAI caused an increase in the total pedometer count at all doses tested( Figure 2E ).

[0221] To evaluate the effect of MEAI on sweet preference, the sucrose preference test (SPT), a commonly used reward-based test for anhedonia, was employed. After a single injection, the 40 mg / kg dose of MEAI significantly reduced the acute preference of mice for the sucrose solution without any accompanying decrease in water intake levels. This effect was most pronounced during the initial 24-hour period, with a slight decrease noted during the subsequent 24-hour period( Figure 3) These results suggest that MEAI has the potential to impede rewarding stimuli, causing a reduction in the hedonic effects typically associated with palatable foods.

[0222] The effects of MEAI on food addiction behavior and the metabolic efficacy of MEAI in regulating appetite, treating obesity, and related abnormalities were evaluated in a DIO mouse model ( Figure 4A ). To assess the effects of long-term exposure, a suboptimal dose of 40 mg / kg / day of MEAI was tested in this model. At baseline prior to drug treatment, mice fed an HFD exhibited significantly greater body weights compared to the control group fed an STD. After a 28-day treatment period, MEAI treatment significantly reduced the body weights of mice fed an HFD ( Figure 4B ), resulting in an approximately 15% reduction in overall body weight compared to the vehicle-treated group of obese mice ( Figure 4C 、D). Thus, MEAI significantly reduced the overweight of mice fed an HFD. In addition, MEAI treatment reduced obesity associated with obesity in the DIO model, maintaining lean body mass, lean body mass rate, and net lean mass ( Figure 4E 、F), while simultaneously reducing total fat mass ( Figure 4G 、H).

[0223] Analysis of feeding behavior revealed that mice fed an HFD in both the MEAI and vehicle-treated groups consumed fewer meals, as indicated by a decrease in food intake per meal ( Figure 5A ). However, cumulative food intake over a 24-hour period was similar in all groups ( Figure 5B 、C), mainly due to the higher caloric density of the HFD ( Figure 5D ). In addition, as found in the acute setting of lean animals ( Figure 1D 、E), MEAI treatment had no effect on water intake in obese mice ( Figure 5E ).

[0224] Metabolically, the RER was slightly reduced in both the HFD vehicle and MEAI-treated groups compared to the STD vehicle group ( Figure 5F ). Compared to the HFD vehicle-treated group, MEAI administration slightly increased oxygen consumption and carbon dioxide production ( Figure 5G 、H). Notably, compared to both the HFD and STD vehicle-treated groups, the MEAI-treated group showed a significant increase in energy expenditure, with a clear elevation observed during both the light and dark phases ( Figure 5I ). Regression analysis of TEE compared to body weight revealed a significant difference between groups independent of body weight ( Figure 5J ). In addition, compared to both the HFD vehicle-treated group and the STD vehicle-treated group, MEAI treatment caused an increase in the total rate of FO ( Figure 5K)。However, CHO was significantly reduced in both the vehicle-treated and MEAI-treated groups, where the drug itself had no effect( Figure 5L )。

[0225] Analysis of locomotor activity showed that MEAI treatment significantly increased spontaneous locomotion throughout the day, such as walking activity and grooming, but was more prominent during the dark phase of the day, consistent with their nocturnal activity nature( Figure 6A )。Interestingly, compared with the HFD-vehicle-treated group, although the spontaneous locomotor activity, speed, and total distance traveled increased in the MEAI group, the drug did not exceed the level of the STD vehicle-treated group, indicating that it did not induce overstimulation( Figure 6B -D). A similar behavioral pattern was observed in the running wheel movement parameters, which are purely spontaneous activities. Animals treated with MEAI demonstrated an increased ability to run on the spontaneous running wheel, and their speed was similar to that of the STD vehicle-treated group( Figure 6E 、F). In addition, analysis of the time spent by mice in the cage engaging in various activities showed that mice treated with MEAI exhibited a preference for spontaneous activities such as running wheel and walking movements, as well as an extended interaction time with the food and water dispensers( Figure 6G )。

[0226] MEAI improves glucose control in DIO mice. Obesity is a well-known contributing factor to insulin resistance and hyperglycemia, which may ultimately lead to the onset of diabetes. In the DIO model, substantial impairment of glucose tolerance and increased hyperinsulinemia were observed, as demonstrated by the results of glucose and insulin tolerance tests. However, after treatment with MEAI, a significant improvement in glucose metabolism was observed( Figure 7A -D), where fasting blood glucose and insulin levels were also reduced( Figure 7E 、F). These beneficial effects of MEAI were reflected in HOMA-IR and ISI( Figure 7G 、H), indicating a positive effect of MEAI on glucose metabolism. In addition, MEAI normalized insulin sensitivity( Figure 7C and D), indicating a positive effect on glucose metabolism.

[0227] Treatment with MEAI improves HFD-induced dyslipidemia. To investigate whether MEAI could alleviate the dyslipidemia commonly associated with obesity, lipid profiles were analyzed. The results showed that treatment with MEAI significantly reduced LDL levels compared with the HFD vehicle group, while HDL levels did not change significantly. This reduction in LDL levels was accompanied by an increase in the HDL-to-LDL ratio, indicating a positive effect on lipid metabolism( Figure 8A -C). In addition, there was a tendency for cholesterol levels to decrease in the MEAI-treated group( Figure 8D), although this change was not statistically significant. However, no significant alteration in circulating triglyceride levels was observed in any of the study groups( Figure 8E ). These findings suggest that MEAI may have a potential therapeutic effect on dyslipidemia associated with obesity.

[0228] Initial tests examining the effect of MEAI on renal function showed normalization of the kidney weight-to-body weight ratio after MEAI administration, accompanied by a slight improvement in blood urea nitrogen (BUN) levels( Figure 9A -C).

[0229] MEAI reverses obesity-induced liver dysfunction and steatosis. Obesity is a well-established risk factor for the development of NAFLD, which is characterized by hepatic steatosis caused by an imbalance between hepatic fatty acid uptake, synthesis, oxidation, and export. Given the promising effects of MEAI on body weight, fat oxidation, and circulating lipid levels, its effect on hepatic steatosis was investigated. Findings demonstrated that MEAI treatment decreased the liver weight( Figure 10A ) and normalized its ratio to body weight( Figure 10B ) in HFD-fed mice. Although MEAI administration did not significantly alter ALT or AST levels compared to the HFD vehicle group, it significantly decreased ALP levels, which may indicate reduced liver injury( Figure 10C -E). In addition, treatment with MEAI had a positive effect on hepatic lipid accumulation, as evidenced by a significant decrease in hepatic triglycerides and a trend towards decreased hepatic cholesterol levels compared to the HFD vehicle control( Figure 10F 、G). A decrease in Oil Red O staining and a reduction in the number of lipid vacuoles in the livers of MEAI-treated mice further supported these findings compared to the HFD vehicle-treated group( Figure 10H 、I). Collectively, these findings suggest that MEAI may have beneficial effects on hepatic lipid accumulation and liver function in the context of obesity-related NAFLD.

[0230] From the detailed description, many features and advantages of the present disclosure are apparent, and thus it is intended to cover all such features and advantages of the present disclosure that fall within the true spirit and scope of the present disclosure by the appended claims. In addition, since many modifications and variations will readily occur to those skilled in the art, it is not desirable to limit the present disclosure to the exact construction and operation shown and described, and accordingly, all suitable modifications and equivalents falling within the scope of the present disclosure may be resorted to.

[0231] In addition, those skilled in the art will understand that the concepts on which the present disclosure is based can easily be used as a basis for designing other structures, methods, and systems for several purposes for implementing the present disclosure. Therefore, the claims should not be regarded as limited by the foregoing description or examples.

Claims

1. A method for treating a metabolic condition in a subject in need thereof, the method comprising administering to the subject a therapeutically acceptable amount of a pharmaceutical composition comprising 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof, wherein the treatment alleviates one or more metabolic syndromes in the subject.

2. The method according to claim 1, wherein the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered in a dose of about 20 mg to about 520 mg.

3. The method according to claim 1, wherein the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered in a dose of about 0.5 mg to about 40 mg.

4. The method according to claim 1, wherein the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered as the following doses: about 20 mg to about 100 mg, about 25 mg to about 90 mg, about 30 mg to about 80 mg, about 40 mg to about 70 mg, or about 50 mg to about 60 mg.

5. The method according to claim 2, wherein the dose is administered as a single dose or as more than one divided dose.

6. The method according to claim 2, wherein the dose is administered as a single dose or as more than one divided dose daily.

7. The method according to claim 2, wherein the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered twice daily.

8. The method according to claim 1, wherein the therapeutically effective amount comprises about 0.0084 mg / kg body weight / day to about 0.67 mg / kg body weight / day, about 0.33 mg / kg body weight / day to about 8.67 mg / kg body weight / day, about 0.33 mg / kg body weight / day to about 1.67 mg / kg body weight / day, about 0.42 mg / kg body weight / day to about 1.5 mg / kg body weight / day, about 0. mg / kg body weight / day to about 1.33 mg / kg body weight / day, about 0.67 mg / kg body weight / day to about 1.17 mg / kg body weight / day, or about 0.83 mg / kg body weight / day to about 1.0 mg / kg body weight / day.

9. The method according to claim 1, wherein the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier and / or excipient.

10. The method according to claim 9, wherein the pharmaceutical composition is a free-flowing powder, tablet, capsule, lozenge, liquid, liquid concentrate, suspension, or syrup.

11. The method according to claim 10, wherein the pharmaceutical composition is a unit dosage form composition.

12. The method according to claim 11, wherein the amount of 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof in the unit dosage form is about 20 mg to about 520 mg, about 0.5 mg to about 40 mg, about 20 mg to about 100 mg, about 25 mg to about 90 mg, about 30 mg to about 80 mg, about 40 mg to about 70 mg, or about 50 mg to about 60 mg.

13. The method according to claim 12, wherein the amount of 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is about 50 mg.

14. The method according to claim 1, wherein the administration of the pharmaceutical composition is oral, sublingual, buccal, vaginal, rectal, parenteral, transdermal, or by inhalation.

15. The method according to claim 14, wherein the parenteral administration is intravenous, intramuscular, or subcutaneous.

16. A method for treating a metabolic condition in a subject in need thereof, the method comprising administering to the subject a therapeutically acceptable amount of a pharmaceutical composition comprising 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof and a pharmaceutical composition comprising an N-acylethanolamine or a pharmaceutically acceptable salt thereof, wherein the treatment alleviates one or more metabolic syndromes in the subject.

17. The method according to claim 16, wherein the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered in a dose of about 20 mg to about 520 mg.

18. The method according to claim 16, wherein the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered in a dose of about 0.5 mg to about 40 mg.

19. The method according to claim 16, wherein the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered in a dose of: about 20 mg to about 100 mg, about 25 mg to about 90 mg, about 30 mg to about 80 mg, about 40 mg to about 70 mg, or about 50 mg to about 60 mg.

20. The method according to claim 17, wherein the dose is administered as a single dose or in more than one divided dose.

21. The method according to claim 17, wherein the dose is administered as a single dose or in more than one divided dose daily.

22. The method according to claim 19, wherein the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is administered twice daily.

23. The method according to claim 16, wherein the therapeutically effective amount of the 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof comprises about 0.0084 mg / kg body weight / day to about 0.67 mg / kg body weight / day, about 0.33 mg / kg body weight / day to about 8.67 mg / kg body weight / day, about 0.33 mg / kg body weight / day to about 1.67 mg / kg body weight / day, about 0.42 mg / kg body weight / day to about 1.5 mg / kg body weight / day, about 0. mg / kg body weight / day to about 1.33 mg / kg body weight / day, about 0.67 mg / kg body weight / day to about 1.17 mg / kg body weight / day, or about 0.83 mg / kg body weight / day to about 1.0 mg / kg body weight / day.

24. The method according to claim 16, wherein the pharmaceutical composition further comprises at least one pharmaceutically acceptable carrier and / or excipient.

25. The method according to claim 24, wherein the pharmaceutical composition is a free-flowing powder, tablet, capsule, lozenge, liquid, liquid concentrate, suspension or syrup.

26. The method according to claim 25, wherein the pharmaceutical composition is a unit dosage form composition.

27. The method according to claim 26, wherein the amount of 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof in the unit dosage form is about 20 mg to about 520 mg, about 0.5 mg to about 40 mg, about 20 mg to about 100 mg, about 25 mg to about 90 mg, about 30 mg to about 80 mg, about 40 mg to about 70 mg or about 50 mg to about 60 mg.

28. The method according to claim 27, wherein the amount of 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof is about 50 mg.

29. The method according to claim 16, wherein the administration of the pharmaceutical composition is oral, sublingual, buccal, vaginal, rectal, parenteral, transdermal or by inhalation.

30. The method according to claim 29, wherein the parenteral administration is intravenous, intramuscular or subcutaneous.

31. The method according to any one of claims 16 - 30, wherein the N-acylethanolamine is selected from N-palmitoylethanolamine (PEA), Me-palmitoylethanolamide (Me-PEA), palmitoylcyclohexylamide, palmitoylbutyramide, palmitoylisopropylamide, oleoylethanolamine (OEA), palmitoylisopropylamide (PIA), salts thereof and any combination thereof.

32. The method according to claim 31, wherein the N-acylethanolamine is palmitoylethanolamide or a pharmaceutically acceptable salt thereof.

33. The method according to any one of claims 16 - 32, wherein the N-acylethanolamine or a pharmaceutically acceptable salt thereof is administered in a dose of about 200 mg to about 1800 mg, about 250 mg to about 1550 mg, about 300 mg to about 1200 mg, about 350 mg to about 950 mg, about 400 mg to about 700 mg, about 450 mg to about 600 mg or about 500 mg to about 550 mg.

34. The method according to claim 33, wherein the dose is administered as a single dose or in more than one divided dose.

35. The method according to claim 33, wherein the dose is administered as a single dose or in more than one divided dose daily.

36. The method according to claim 35, wherein the N-acylethanolamine or a pharmaceutically acceptable salt thereof is administered twice daily.

37. The method according to claim 16, wherein the therapeutically effective amount of the N-acylethanolamine or a pharmaceutically acceptable salt thereof comprises from about 2.5 mg / kg body weight / day to about 36.0 mg / kg body weight / day, from about 3.12 mg / kg body weight / day to about 31.0 mg / kg body weight / day, from about 3.75 mg / kg body weight / day to about 24.0 mg / kg body weight / day, from about 4.38 mg / kg body weight / day to about 19.0 mg / kg body weight / day, from about 5.0 mg / kg body weight / day to about 14.0 mg / kg body weight / day, from about 5.62 mg / kg body weight / day to about 12.0 mg / kg body weight / day, or from about 6.25 mg / kg body weight / day to about 11.0 mg / kg body weight / day.

38. The method according to any one of claims 16 - 37, wherein the N-acylethanolamine is administered simultaneously with the 5-methoxy-2-aminoindane.

39. The method according to claim 38, wherein the N-acylethanolamine and the 5-methoxy-2-aminoindane are administered in a single pharmaceutical composition.

40. The method according to claim 1 or 16, wherein the administration is by oral, mucosal, nasal, sublingual, inhalation, topical, rectal, vaginal, or parenteral route.

41. The method according to claim 40, wherein the parenteral administration is intravenous, intramuscular, or subcutaneous.

42. The method according to any one of claims 1 - 41, wherein treating metabolic syndrome involves one or more of the following: lowering blood pressure, lowering blood glucose, reducing body fat around the waist, normalizing abnormal cholesterol or triglyceride levels, reducing obesity, reducing overweight, reducing body weight, increasing lean mass, reducing fat mass, reducing obesity, increasing energy expenditure, improving glycemic control, reducing hepatic steatosis, reducing glucose uptake, reducing food intake, maintaining glucose homeostasis, reducing dyslipidemia, or maintaining liver function.

43. The method according to claim 42, wherein improving glycemic control involves one or more of improving glucose metabolism, lowering fasting blood glucose levels, or lowering insulin levels.

44. The method according to claim 42, wherein increasing energy expenditure involves one or more of increasing oxygen consumption and carbon dioxide emission, increasing fat oxidation, or increasing spontaneous activity.

45. The method according to claim 42, wherein treating metabolic syndrome reduces obesity.

46. The method according to claim 45, wherein treating metabolic syndrome involves reducing overweight associated with obesity.

47. The method according to claim 45 or 46, wherein treating metabolic syndrome maintains the lean mass of the subject.

48. The method according to claim 45 or 46, wherein treating metabolic syndrome reduces the fat mass of the subject.

49. The method according to claim 45 or 46, wherein treating metabolic syndrome reduces obesity in the subject.

50. The method according to claim 45 or claim 46, wherein treating metabolic syndrome increases energy expenditure.

51. The method according to claim 50, wherein the food consumption is unchanged.

52. The method according to claim 59, wherein the fat utilization is increased.

53. The method according to claim 50, wherein the spontaneous activity is normalized without over-stimulatory effects.

54. The method according to claim 45 or claim 46, wherein treating metabolic syndrome improves blood glucose control.

55. The method according to claim 45 or claim 46, wherein treating metabolic syndrome reverses hyperglycemia, glucose intolerance or hyperinsulinemia.

56. The method according to claim 45 or claim 46, wherein treating metabolic syndrome improves hepatic steatosis.

57. The method according to claim 56, wherein improving hepatic steatosis involves reducing one or more of hepatic lipid accumulation, hepatic triglyceride levels or hepatic cholesterol levels.

58. The method according to claim 45 or claim 46, wherein treating metabolic syndrome maintains glucose homeostasis.

59. The method according to claim 58, wherein maintaining glucose homeostasis involves increasing one or more of glucose tolerance, attenuating insulin resistance, reducing dyslipidemia or reducing hepatic lipid accumulation.

60. Use of a pharmaceutical composition comprising 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof for treating a metabolic condition as described in any one of claims 1-59.

61. Use of a pharmaceutical composition comprising 5-methoxy-2-aminoindane or a pharmaceutically acceptable salt thereof and an N-acylethanolamine (such as palmitoylethanolamide) or a pharmaceutically acceptable salt thereof for treating a metabolic condition as described in any one of claims 16-59.

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