Oral pharmaceutical immediate release composition and method of treatment for weight loss

Abstract

Disclosed is an oral pharmaceutical composition for treating a variety of diseases, comprising a denatonium cation salt and an acid anion selected from the group consisting of acetate (DA), citrate (DC), tartrate (CT), maleate (DM) and combinations thereof (collectively referred to as "denatonium salt") and a pharmaceutical excipient for gastric release of the denatonium salt. Further disclosed is an oral immediate release pharmaceutical composition for substantially releasing an API (active pharmaceutical ingredient) in the gastric region for a gastrointestinal formulation, wherein the API comprises an effective amount of denatonium salt. Preferably, the oral immediate release pharmaceutical formulation comprises about 0.5 g to about 5 g of denatonium salt, delivering a daily dose of about 20 mg to about 150 mg of denatonium salt to an adult.

Description

Technical Field

[0001] The present disclosure provides an oral pharmaceutical composition for treating a variety of diseases, comprising a denatonium cation salt and an acid anion selected from the group consisting of acetate (DA), citrate (DC), tartrate (CT), maleate (DM) and a combination thereof (collectively referred to as "denatonium salt") and a pharmaceutical excipient for gastric release of the denatonium salt. The present disclosure further provides an oral immediate release pharmaceutical composition for substantially releasing an API (active pharmaceutical ingredient) in the gastric region of a gastrointestinal formulation, wherein the API comprises an effective amount of the denatonium salt. Preferably, the oral immediate release pharmaceutical formulation comprises about 0.5 g to about 5 g of the denatonium salt, delivering a daily dose of about 20 mg to about 150 mg of the denatonium salt to an adult. Background Art

[0002] Chemosensory signaling of nutrients plays a role in regulating appetite, digestion, and metabolism. In particular, multiple G protein-coupled receptors (GPCRs) of the bitter taste receptor (TAS2R) family are found not only in the oral cavity, but also on enteroendocrine cells, human gastric smooth muscle cells, adipocytes, and at sites in the chemoreceptor trigger zone in the medulla of the brain.

[0003] Obesity is a global epidemic that has serious health and socioeconomic consequences for millions of adults and children (Bluher, Nat. Rev. Endocrinol. 15 (2019) 288-298). Globally, at least 13% of adults and 7% of children are obese, but in several countries, the prevalence of obesity is at least 30% of the total population (Ng et al., Lancet 384 (2014) 766-781).

[0004] The ideal treatment for obesity is diet and physical exercise, but the observed success rate of such programs is low, around 20%. This is often due to the strong appetite drive, which has redundant pathways for stimulation and is difficult to overcome, as suppressing one appetite-producing pathway often leads to upregulation of compensatory alternative pathways, which induce hunger over time. The various drugs that are already commercially available are generally mediocre in effectiveness, or have associated risks and side effects that many people consider intolerable, or both.

[0005] Anorectic stimulant compounds such as ephedrine, fenfluramine, and dexfenfluramine have been withdrawn from the market due to associated cardiovascular safety risks. Drugs that interfere with nutrient absorption such as the lipase inhibitor orlistat, which blocks fat processing in the intestine, lead to oily stools and diarrhea. CNS-targeted drugs such as sibutramine (a monoamine oxidase inhibitor), rimonabant (a cannabinoid receptor antagonist), etc. have significant central nervous system (CNS) "off-target" effects, often resulting in unintended psychiatric or neurological manifestations.

[0006] Behavioral interventions for obesity such as exercise programs and dietary changes often fail, and bariatric surgery is not an option for most people. Anti-obesity drugs can effectively reduce body weight; however, they are associated with side effects ranging from headaches, nausea, and dizziness to serious psychiatric and cardiovascular events (MO Dietrich et al., Nat. Rev. Drug Discov. 11 (2012) 675-691). Given the enormous medical, social, and economic burden of obesity, there is an urgent need to develop new, safe, and effective therapeutics for this debilitating and potentially fatal disease.

[0007] Bitter taste receptors (TAS2Rs) comprise a family of several G protein-coupled receptors (GPCRs) expressed on the tongue as well as other organs including the brain, oral cavity, lung, pancreas, and gastrointestinal mucosa (Jaggupilli et al., Mol. Cell. Biochem. 426 (2017) 137-147).

[0008] Denatonium benzoate activates eight human TAS2Rs (TAS2R 4, 8, 10, 13, 39, 43, 46, and 47) to varying degrees (Meyerhof et al., Chem. Senses 35 (2010) 157-170). In rodent obesity models, denatonium benzoate suppresses food intake and inhibits weight gain (Avau et al., PLoS One 10 (2015) e0145538; and Glendinning et al., Physiol. Behav. 93 (2008) 757-765). In addition, in healthy volunteers, denatonium benzoate attenuated intragastric gastric motility, reduced nutrient tolerance, reduced hunger ratings, and increased postprandial satiety after intragastric administration (Avau et al., Sci. Rep. 5 (2015) 15985; and Deloose et al., Am. J. Clin. Nutr. 105 (2017) 580-588). However, due to the disgusting nature of denatonium benzoate, two studies of denatonium benzoate presented a significant side effect problem. Therefore, there is a significant need in the art to address obesity and related disorders with safer improved bitter agonists. The present disclosure addresses this need. Summary of the invention

[0009] The present disclosure is based on the discovery that denatonium salts with acid-tasting anions have a better side effect profile seen in comparative in vivo studies compared to denatonium benzoate, the only available denatonium salt and the denatonium salt reported in earlier studies.

[0010] The present disclosure provides an oral pharmaceutical composition for treating a variety of diseases, comprising a denatonium cation salt and an acid anion selected from the group consisting of acetate (DA), citrate (DC), tartrate (CT), maleate (DM) and a combination thereof (collectively referred to as "denatonium salt") and a pharmaceutical excipient for gastric release of the denatonium salt. The present disclosure further provides an oral immediate release pharmaceutical composition for substantially releasing an API (active pharmaceutical ingredient) in the gastric region of a gastrointestinal formulation, wherein the API comprises an effective amount of the denatonium salt. Preferably, the oral immediate release pharmaceutical formulation comprises about 0.5 g to about 5 g of the denatonium salt, delivering a daily dose of about 20 mg to about 150 mg of the denatonium salt to an adult.

[0011] The present disclosure provides an oral pharmaceutical immediate gastric release pharmaceutical formulation ("oral formulation") comprising particles comprising a denatonium cation salt and an acid anion selected from the group consisting of acetate (DA), citrate (DC) tartrate (CT), maleate (DM) and a combination thereof (collectively referred to as "denatonium salt") and a pharmaceutical excipient for gastric release of the denatonium salt. Preferably, the pharmaceutical excipient comprises talc, cellulose and sugar. Preferably, the oral formulation further comprises an organic acid selected from the group consisting of acetic acid, malic acid, maleic acid, citric acid and a combination thereof. Preferably, the oral formulation further comprises about 0.5 g to about 5 g of acetic acid. More preferably, the daily dose of acetic acid for adults is about 1.5 g to about 3 g. Preferably, the daily dose of DA for adults is about 10 mg to about 600 mg or about 5 mg / kg to about 50 mg / kg body weight per day. More preferably, the daily dose of DA for adults is about 10 mg to about 200 mg. Most preferably, the daily dose of DA for adults is about 10 mg to about 100 mg, or for achieving a concentration of about 10 ppb to about 10 ppm in the gastrointestinal tract. In view of the sustained release or immediate release characteristics, the daily dose of DA is once a day, twice a day or three times a day.

[0012] Further, the present disclosure provides a sustained release oral formulation comprising DA and acetic acid powder in a sustained release cellulose and mannitol excipient formulation. Preferably, the daily dose of DA for adults is about 10 mg to about 600 mg. More preferably, the daily dose of DA for adults is about 10 mg to about 200 mg. Most preferably, the daily dose of DA for adults is about 10 mg to about 100 mg, or for achieving a concentration of about 10 ppb to about 10 ppm in the gastrointestinal tract. Preferably, the oral formulation comprises about 0.01% to about 10 wt % DA and about 10% to about 90 wt % dry acetic acid powder. Preferably, the dosage of DA administered is about 500 nmol / kg to about 4 μmol / kg. Preferably, the dosage of DA administered to adults is about 10 mg to about 50 mg. In view of the release characteristics, the daily dose of DA is once a day, twice a day or three times a day.

[0013] The present disclosure further provides a method for achieving weight loss, comprising administering an oral pharmaceutical immediate gastric release pharmaceutical formulation ("oral formulation") comprising particles comprising a denatonium cation salt and an acid anion selected from the group consisting of acetate (DA), citrate (DC) tartrate (CT), maleate (DM) and combinations thereof (collectively referred to as "denatonium salt") and a pharmaceutical excipient for gastric release of the denatonium salt. Preferably, the pharmaceutical excipient comprises talc, cellulose and sugar. Preferably, the oral formulation further comprises an organic acid selected from the group consisting of acetic acid, malic acid, maleic acid, citric acid and combinations thereof. Preferably, the oral formulation further comprises about 0.5 g to about 5 g of acetic acid. More preferably, the daily dose of acetic acid for adults is about 1.5 g to about 3 g. Preferably, the daily dose of DA for adults is about 10 mg to about 600 mg or about 5 mg / kg to about 50 mg / kg body weight per day. More preferably, the daily dose of DA for adults is about 10 mg to about 200 mg. Most preferably, the daily dose of DA for adults is from about 10 mg to about 100 mg, or to achieve a concentration of from about 10 ppb to about 10 ppm in the gastrointestinal tract. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Comparison of a 56-day DIO mouse weight loss study (Example 3) showing body weights on the indicated days. The highest dose DA group (23.1 mg / kg) showed the lowest mean body weight.

[0015] Figure 2 Results are shown for body weight changes from the 56-day study in Example 3. Animals treated with 23.1 mg / kg DA showed the lowest weight gain relative to the high dose DB group.

[0016] Figure 3 Results are shown for serum insulin at the end of the 56-day study in Example 3. Serum insulin in the 23.1 mg / kg DA group was close to baseline values ​​(ie, before treatment) and was significantly lower compared to the vehicle-treated group.

[0017] Figure 4 It was shown that there was no statistical difference in serum HBAlc levels among all experimental groups in Example 3.

[0018] Figure 5 Cumulative food consumption over 24 hours for the one-day rat study described in Example 4 is shown.

[0019] Figure 6 Shown are the mean absolute body weight changes during the 56 day treatment period in DIO mice from Example 6.

[0020] Fig. 7A and 7BDose-lethal curves for DA and DB from Example 7 are shown.

[0021] Figure 8 Display drug product / formulation flow chart. DETAILED DESCRIPTION

[0022] The present disclosure is based on a surprising discovery that the anti-obesity effects of denatonium salts with acidic anions (acetate, citrate, tartrate and maleate) are excellent (both safety and efficacy) using in vitro and in vivo obesity models. The aim of our study was to determine the effects of denatonium salts with acidic anions on food and water consumption, weight control.

[0023] In a short-term food intake inhibition study, the doses of DA administered in Sprague Dawley rats were 7.5, 15, 30, and 60 pmol / kg. The corresponding human equivalent doses (HED) were 1.2, 2.4, 4.9, and 9.7 pmol / kg, respectively. In a long-term food intake inhibition study, the dose of DA was 60 pmol / kg in C57BL / 6NTac mice. The corresponding HED was 4.9 pmol / kg. As background, according to Avau et al., Sci. Rep. 5 (2015) 15985), oral administration of only 60 pmol / kg (26.8 mg / kg) of denatonium benzyl (DB), a salt related to DA, significantly inhibited gastric emptying rate in normal C57BL / 6 mice. In another study, once daily treatment with 60 pmol / kg DB (26.8 mg / kg) induced weight loss in C57BL / 6DIO mice over a 28-day period compared to vehicle. According to Avau et al., healthy volunteers receiving 1 pmol / kg DB showed reduced nutrient tolerance and increased satiety. Thus, the disclosed formulations provide a DA dose of about 500 nmol / kg to about 10 pmol / kg, which corresponds to about 10 mg to about 230 mg for adults.

[0024] Table 1: Denatonium salts

[0025]

[0026] Denatonium Benzyl (DB)

[0027] IUPAC name: Benzyl-[2-(2,6-dimethylanilino)-2-oxoethyl]-diethylammonium benzoate Molecular formula: C28H34N2O3

[0028] Molecular weight:446.581g / mol

[0029] CAS No.:3734-33-6

[0030] ChemSpider ID:

[0031] Denatonium, commonly available as denatonium benzoate (trade names such as BITTERANT-b, BITTER+PLUS, Bitrex, or Aversion). It is used as an aversive (bittering) agent to prevent inappropriate ingestion. Denatonium benzoate is used in denatured alcohol, antifreeze, nail biting agents, respiratory mask fit testing, animal repellents, liquid soaps, and shampoos. It is not known to pose any long-term health risks.

[0032] Therapies that can trigger extraoral bitter taste receptors in the gut, brain, and other areas such as adipocytes using compounds with low intrinsic toxicity offer a relatively safe means to selectively reduce appetite and increase satiety without the "off-target" CNS effects or gastrointestinal disturbances typical of other obesity drugs.

[0033] In addition to obesity, one clinical use of oral ingestion of DA-containing tablets or pills in combination with an organic acid, such as acetic acid, is Prader-Willi syndrome. Among the key features of this genetic disorder are a persistent hunger drive and a lack of satiety even after eating large amounts of food. Accordingly, the present disclosure provides a method for treating Prader-Willi syndrome (PWS) comprising an anti-obesity oral formulation comprising (a) denatonium acetate (DA), (b) an organic acid selected from the group consisting of acetic acid, malic acid, maleic acid, citric acid, and combinations thereof, and (c) a pharmaceutical excipient that promotes sustained release during transit through the gastrointestinal tract. Example

[0034] Example 1

[0035] This example describes a method for formulating Denatonium Acetate / Acetic Acid Release Tablets, 44.6 mg / 500 mg.

[0036] Table 2

[0037]

[0038] Microcrystalline cellulose (Avicel PH101), denatonium acetate, PVP 30 (half quantity) and mannitol were added to a 10 cubic foot V blender and mixed for 10 minutes. The mixture was transferred to a high shear granulator and granulation was started with a controlled spray of acetic acid (half quantity) at a rate of 800 g / min. After granulation, the wet granules were removed and placed in a tray dryer controlled at 50°C for a period of time until the final moisture content was less than 2% w / w. The dried granules were then passed through a Fitzmill equipped with an 18 mesh screen. The milled granules were then placed back into the same high shear granulator and the remaining half of PVP 30 was added and granulated again with the remaining half of acetic acid. The wet granules were removed and dried at 50°C until the moisture content was less than 2%. The dried granules were milled in a Fitzmill with an 18 mesh screen and then mixed with magnesium stearate for 5 minutes in a 10 cubic foot V blender and the final mixture (uncoated tablets) was compressed in a tablet press with a target weight of 786.6 mg and a hardness of 10 kp.

[0039] The coating solution was prepared by dispersing dibutyl sebacate in Aquacoat ECD 30 dispersion and gently mixing for 1 hour. Uncoated tablets were loaded in a pan coater and sprayed with the coating solution at a controlled spray rate of 80 g / min. Drying was continued for 30 minutes after the coating was completed.

[0040] Example 2

[0041] This example describes a method for formulating Denatonium Acetate / Acetic Acid Immediate Release Tablets, 22.3 mg / 250 mg.

[0042] Table 3

[0043] Components <![CDATA[Per dose 2 , mg]]> Quantity,kg Denatonium acetate 22.3 4.46 Acetic acid, NF (36.5% w / w) 685 137 Microcrystalline Cellulose 100 20 Mannitol 90.2 18.04 Polyvinylpyrrolidone 30 (PVP 30) 25 5 Magnesium Stearate 2.5 0.5

[0044] 2 One dose can come from 1-5 tablets.

[0045] Microcrystalline cellulose (Avicel PH101), denatonium acetate, PVP 30 (half the quantity) and mannitol were added to a 10 cubic foot V blender and mixed for 10 minutes. The mixture was transferred to a high shear granulator and granulation was started with a controlled spray of acetic acid (half the quantity) at a rate of 800 g / min. After granulation, the wet granules were removed and placed in a tray dryer controlled at 50°C for a period of time until the final moisture content was less than 2% w / w. The dried granules were then passed through a Fitzmill equipped with an 18 mesh screen. The milled granules were then placed back into the same high shear granulator and the remaining half of the PVP 30 was added and granulated again with the remaining half of the acetic acid. The wet granules were removed and dried at 50°C until the moisture content was less than 2%. The dried granules were milled in a Fitzmill with an 18 mesh screen and then mixed with magnesium stearate for 5 minutes in a 10 cubic foot V blender and the final mixture was compressed in a tablet press with a target weight of 500 mg and a hardness of 10 kp.

[0046] Example 3

[0047] This example shows acute and chronic in vivo studies comparing the weight loss properties of two salts, DA and DB (denatonium benzyl), with the same cation and different anions. The 56-day study determined the behavioral effects of the bitter taste receptor agonist denatonium acetate (DA) compared to denatonium benzyl (DB) in a diet-induced obesity (DIO) mouse model. Animals were acclimated to life for at least 3 days, maintained on a standard chow diet, 12:12 light / dark cycle and housed in groups of 2-3 in stacked filtered cages. The study duration was a 3-5 day acclimation period + a 28 day study period and a 2-3 day test period after the study. There were two DA doses, 2.9 and 23.1 mg / kg BID (3.1 and 60 μmol / kg BID), 26.8 mg / kg DB BID and a distilled water control vehicle, DA and DB were formulated in distilled water. Mice were C57BL / 6NTad mice at least 12 weeks old and 15 mice per group (low-dose DA, high-dose DA, high-dose DB and control). There were daily gross observations, and body weight measurements for each animal on days 0, 1, 4, 7, 9, 11, 14, 16, 18, 21, 23, 25, 28, 30, 32, 34, 36, 39, 41, 43, 46, 48, 50, 53, and 56. Food intake was measured on days 0, 7, 14, 21, 28, 35, 42, 49, and 56. Metabolic biomarkers (blood glucose, blood insulin, blood HbA1c) were measured at the beginning and end of the study. DA, DB, or distilled water control were administered orally (PO) at a volume of 5 mL / kg body weight.

[0048] Results are shown in Figure 1-4 , showing that high-dose DA outperformed high-dose DB in terms of weight loss improvement.

[0049] Example 4

[0050] This example provides a 24 hour study comparing DA to DB in rats (male Sprague Dawley, Charles River) over a 24 hour period. Five groups of 15 rats each were QID gavaged with distilled water for vehicle control, QID gavaged with DB at a dose of 26.8 mg / kg, QD gavaged with DA low dose 2.9 mg / kg, and QD gavaged with DA high dose 23.1 mg / kg. Food intake was measured 2 hours, 4 hours, 6 hours, 8 hours, and 24 hours after administration. The results of cumulative food consumption over a 24 hour period are shown in Figure 5 , drug treatment had a significant main effect on cumulative food consumption, with the high-dose DA group having the largest effect.

[0051] Example 5

[0052] This example describes the synthesis of denatonium acetate (DA).

[0053] Step 1: Synthesis of denatonium hydroxide from lidocaine

[0054] Add 25 g of lidocaine, 60 ml of water and 17.5 g of benzyl chloride to a reflux apparatus under stirring and heating at 70-90°C. Heat and stir the solution for 24 hours before setting. Cool the solution to 30°C. Remove unreacted reagents with 3x 10 mL of toluene. Dissolve 65 g of sodium hydroxide in 65 mL of cold water under stirring and add it to the aqueous solution under stirring over a period of 3 hours. Filter the mixture, wash with some water and dry in the open air. Recrystallize in hot chloroform or hot ethanol.

[0055]

[0056] Step 2: Preparation of denatonium acetate from denatonium hydroxide

[0057] 10 g of denatonium hydroxide (MW: 342.475 g / mol, 0.029 mol), 20 mL of acetone, and 2 g of glacial acetic acid (0.033 mol) dissolved in 15 mL of acetone were added to a reflux apparatus, and the mixture was stirred and heated to 35° C. for 3 hours. Then evaporated to dryness and recrystallized in hot acetone.

[0058]

[0059] Example 6

[0060] This example compares the efficacy of DA with DB in terms of food suppression and weight control. As background, according to Avauet al., Sci. Rep. 2015; 5: 15985, oral administration of 26.8 mg / kg DB significantly inhibited gastric emptying rate in normal C57BL / 6 mice. In our first in vivo study, 45 male SD rats (purchased from Envigo, 8-10 weeks old) were divided into three groups (15 rats per group) and administered a single oral dose of vehicle (distilled water), 26.8 mg / kg DB, or 23.1 mg / kg DA, respectively, with a 24-hour observation period to compare the efficacy of DB with DA in reducing food intake.

[0061] Table 4: Mean cumulative food intake during the 24-hour observation period

[0062]

[0063] The mean cumulative food intake during the 24-hour observation period is presented in Table 4. Administration of DB or DA reduced cumulative food intake during all indicated time intervals compared to vehicle. In addition, a greater reduction in food intake was observed for doses of 23.1 mg / kg DA than for 26.8 mg / kg DB, despite the fact that the molar dose of DA was even lower than that of DB (57.4 pmol / kg vs. 60 pmol / kg). Thus, these data show that DA has a greater efficacy than DB in terms of food intake reduction based on the different anions of the salt.

[0064] In another published paper, once daily treatment with 26.8 mg / kg DB induced weight loss in C57BL / 6 diet-induced obese (DIO) mice over a 28-day period compared to vehicle (Avau et al., PLoS One 2015; 10(12): e0145538). In a second in vivo study, 45 male C57BL / 6 DIO mice (purchased from Envigo, 18 weeks old, fed a high-fat diet) were divided into three groups (15 mice each) and orally administered with vehicle (distilled water), 26.8 mg / kg DB, or 23.1 mg / kg DA twice daily (BID) for a 56-day treatment period to compare the efficacy of DB with DA in terms of food intake reduction and body weight control. Briefly, food weight was recorded once a week for each cage at 0 hours and then 24 hours later, allowing calculation of food consumption for that 24-hour interval. In addition, mice were weighed three times a week (every 2-3 days) starting from day 0.

[0065] The mean food consumption per animal for 24-hour intervals on the indicated measurement days during the treatment period is shown in Table 5. Of note, mice dosed with 23.1 mg / kg DA exhibited nominally reduced food consumption compared to vehicle-dosed mice; this effect was seen throughout the study. Lower food consumption was also seen in animals dosed with 26.8 mg / kg DB on days 0, 7, 28, 35, 42, and 49 (compared to vehicle-dosed mice), but not on days 14, 21, and 56. Moreover, food consumption in animals treated with 23.1 mg / kg DA was less than that in mice treated with 26.8 mg / kg DB on all indicated measurement days except day 42.

[0066] Table 5: Mean food consumption per animal in 24-hour intervals on the indicated measurement days

[0067]

[0068] The mean absolute weight change (in grams) and standardized weight change (% of baseline) for the three treatment groups during the 56-day treatment period are presented in Figure 6 and Table 6.

[0069]

[0070] Feeding with a high-fat diet induced weight gain in all three experimental groups. However, treatment with 26.8 mg / kg DB or 23.1 mg / kg DA resulted in less weight gain compared with vehicle treatment. In particular, weight gain in animals treated with 23.1 mg / kg DA was less than that in animals treated with 26.8 mg / kg DB from day 34 to day 56. Based on these data, DA has a stronger effect than DB not only in terms of food intake reduction but also in terms of body weight control based on the different anions of the salt.

[0071] Example 7

[0072] This example shows the maximum tolerated dose of two denatonium salts, commercially available denatonium benzyl (DB, molecular weight: 446.58 g / mol) and denatonium acetate (monohydrate) (DA, molecular weight (MW): 402.53 g / mol) synthesized under GMP conditions by Aardvark Therapeutics under supply contract. The drugs were administered to Sprague Dawley rats with a 14-day observation period. 24 male Sprague Dawley (SD) rats and 24 female SD rats were purchased from Envigo, 8-10 weeks old. The DA group had four dose levels (120, 360, 1000, and 2000 mg / kg, administered by oral gavage as a single dose), 3 rats per sex, and a total of 6 animals per dose level; the DB group had four dose levels (120, 360, 1000, and 2000 mg / kg, administered by oral gavage as a single dose), 3 rats per sex, and a total of 6 animals per dose level. The estimated median lethal dose (LD50) was determined by calculating the LD50 by nonlinear regression [model: Y=100 / (1+10^(LogEC50-X)), Hill slope=1.0]. The mortality rates for all dose levels in both experimental groups are presented in Table 7.

[0073] Table 7: Mortality at all dose levels for DA and DB

[0074]

[0075]

[0076] Although the MTD (maximum tolerated dose) of DA and DB in rats is the same (360 mg / kg), administration of 1000 mg / kg DA resulted in a lower mortality rate than the same dose of DB (50% vs. 66.7%). Therefore, these data show that DA is a safer drug than DB based on the different anions of the salts.

[0077] The dose-mortality curves for DA and DB are shown in Fig. 7A and 7B The estimated LD50 values ​​and fitting parameters for DA and DB are presented in Table 8. The estimated LD50 for DA was higher than that for DB (945 mg / kg vs. 784 mg / kg), with similar goodness-of-fit parameters.

[0078] Table 8: Estimated LD50 and fitting parameters

[0079]

[0080] Therefore, based on the different anions of the salts, DA is a safer drug than DB.

[0081] Example 8

[0082] This example provides an immediate release 50 mg granule formulation of denatonium acetate monohydrate (DA) as the free base as an immediate gastric release oral pharmaceutical formulation.

[0083]

[0084] A schematic diagram of the formulation process is shown in Figure 8 .

[0085] The detailed fabrication steps are described as follows.

[0086] 1. Drug layering process - drug layered granulation

[0087] The drug layering process was carried out in a fluidized bed granulator (rotor granulator) equipped with a rotor insert. The drug solution was prepared by dissolving Kollidon 30 and denatonium acetate in ethanol. The drug solution was sprayed tangentially onto a bed of sugar spheres (35 / 45 mesh) moving in a circular motion in the rotor granulator. The final drug loaded granules were then dried in the rotor granulator for 10 minutes, removed and screened through a #20 mesh.

[0088] 2. Seal coating process - seal coating pellets

[0089] The seal coating dispersion was prepared by dissolving Hypromellose E5 separately in a mixture of ethanol and purified water (1:1) until a clear solution was obtained. The remaining amount of ethanol was then added to the above solution followed by talc. The dispersion was mixed for 20 minutes to allow the talc to be evenly dispersed. The seal coating dispersion was tangentially sprayed onto the drug loaded granules to achieve a weight gain of 5%. The seal coating granules were then dried in a rotor granulator for 5 minutes, removed and further dried in a tray dryer / oven at 55°C for 2 hours. The seal coating granules were then screened through a #20 mesh.

[0090] 3. Final Mixing - Denatonium Immediate Release (IR) Granules

[0091] The seal coating granules were mixed with talc sieved through a #60 mesh using a V-mixer for 10 minutes and taken out. The mixed seal coating beads (Denatonium IR granules) were used for capsule filling.

[0092] 4. Capsules - Denatonium capsules, 50 mg

[0093] 50 mg of Denatonium IR granules were filled into No. 1 white opaque hard gelatin capsules using an automatic capsule filling machine. The capsules were then passed through an online capsule polisher and metal detector. In-process control of capsule weight and appearance was implemented during the capsule filling process. Acceptable quality limit (AQL) sampling and testing of composite samples was implemented by Quality Assurance (QA) during the capsule filling process. Finished composite samples were collected and analyzed as per release test specifications.

[0094] 5. Packaging - capsule, 50mg-30 capsules

[0095] Thirty 50 mg capsules were packaged into 50 / 60 cc white HDPE round S-line bottles with 33 mm white CRC caps. The bottles were twisted and sealed using an induction sealer.

Claims

1. Use of denatonium acetate in the preparation of a medicament for achieving weight loss.

2. The use of claim 1, wherein the denatonium acetate is in granules comprising the denatonium acetate and a pharmaceutical excipient for gastric release of the denatonium acetate.

3. The use of claim 2, wherein the pharmaceutical excipient comprises talc, cellulose and sugar.

4. The use of claim 2, wherein the particles are in an oral pharmaceutical immediate gastric release pharmaceutical formulation.

5. The use of claim 4, wherein the formulation releases an effective amount of the denatonium acetate in the gastric region of the gastrointestinal tract.

6. The use of claim 4, wherein the formulation comprises 0.5 g to 5 g of the denatonium acetate.

7. The use of claim 4, wherein the formulation further comprises an organic acid selected from the group consisting of acetic acid, malic acid, maleic acid, citric acid and combinations thereof.

8. The use of claim 4, wherein the formulation further comprises 0.5 g to 5 g of acetic acid.

9. The use of claim 1, wherein the daily dose of the denatonium acetate for adults is 10 mg to 600 mg.

10. The use of claim 1, wherein the daily dose of the denatonium acetate for adults is 5 mg / kg to 50 mg / kg body weight per day.

11. The use of claim 1, wherein the daily dose of the denatonium acetate for adults is 10 mg to 200 mg.

12. The use of claim 1, wherein the daily dose of the denatonium acetate for adults is 10 mg to 100 mg.

13. The use of claim 1, wherein the denatonium acetate achieves a concentration of 10 ppb to 10 ppm in the gastrointestinal tract at a daily dose for adults.

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