EA040171B1 - PHARMACEUTICAL COMPOSITION FOR TREATMENT OF OBESITY AND ITS ASSOCIATED DISORDERS AND METHOD FOR ITS PRODUCTION - Google Patents

Description

Technical field

The present invention relates to a pharmaceutical composition/composition for use in the prevention of dietary obesity. More specifically, the invention relates to a composition/composition containing a synergistic combination of oleoylethanolamide (OEA) and other natural ingredients, for the treatment of obesity and related disorders associated with fat metabolism, such as non-alcoholic steatohepatitis (NASH) and non-alcoholic fatty liver disease (NAFLD) . It may also be helpful in the fight against obesity by maintaining body weight. The invention also provides various compositions and methods for their preparation.

State of the art

Obesity has become one of the most significant public health problems currently facing the world. Obesity is a serious health problem involving numerous factors. Experience shows that obesity also involves control of appetite regulation and energy metabolism. In the United States, the incidence of obesity has risen by 32% among adults and 40% among children over the past two decades. Obesity is not just about being overweight. It is a metabolic disorder caused by the accumulation of excess dietary calories in visceral fat and the release of high concentrations of free fatty acids into various organs. As defined by the World Health Organization (WHO), a body mass index (BMI) above 25 is classified as overweight and a BMI above 30 as obese. According to general statistics, the population of overweight and obese individuals is approximately over 4.5 billion, 27% of whom are obese.

The increasing incidence of obesity is an international concern as it increases the risk of various metabolic disorders and diseases such as insulin resistance, type 2 diabetes, hyperlipidemia, coronary heart disease, steatosis, cardiovascular disease, diabetes, chronic lower respiratory disease, chronic hepatitis and cirrhosis of the liver, hypertension, kidney failure, as well as NASH, NAFLD and certain types of cancer. Obesity not only increases mortality and causes serious medical complications, but also affects daily life.

The incidence of NAFLD has increased in parallel with the emergence of central obesity and is now the most common fatty liver disease in developed countries. NAFLD is defined as the presence of hepatic steatosis with or without inflammation and fibrosis, in the absence of prior evidence of alcohol intake. NAFLD is subdivided into NAFLD and NASH. In NAFLD, hepatic steatosis is present without evidence of significant inflammation, whereas in NASH, hepatic steatosis is associated with liver inflammation that may be histologically indistinguishable from alcoholic steatohepatitis.

The histological spectrum of NAFLD includes the presence of steatosis per se, fatty liver, and inflammation. NASH is a more serious chronic liver disease characterized by excessive accumulation of fat in the liver, which includes chronic inflammation leading to progressive fibrosis (scarring) that can lead to cirrhosis, hepatocellular carcinoma (HCC), subsequent liver failure, and death (Brunt et al ., Am J Gastroenterol 94:2467-2474, 1999; Brunt, Semin Liver Dis 21:3-16, 2001; Takahashi Y et al., World J Gastroenterol 18:2300-2308, 2012).

NAFLD is generally recognized as being associated with metabolic syndrome such as insulin resistance, type 2 diabetes mellitus, hyperlipidemia, hypertension, and obesity. It usually occurs in people who are overweight or obese. NAFLD affects 30% of the world's population and approximately 80% of obese people. NAFLD is more common in women and the more common cause is obesity. Recent studies have also shown that overweight and obesity have been clearly and progressively associated with increased occurrence of NAFLD.

NASH is a progressive, acute form of NAFLD. Over a recent period of 10 years, up to 20% of patients with NASH have been diagnosed with cirrhosis of the liver, and approximately 10% have had a fatal outcome due to liver disease. NASH progresses to cirrhosis in approximately 1520% of affected individuals and is currently one of the leading rates for liver transplantation in the United States. There is currently no proven treatment for NASH.

Despite the continuous increase in the incidence of obesity associated with chronic liver diseases such as NAFLD and NASH in all developed countries, limited pharmacological drugs are currently available to the public to treat obesity or its comorbidities in an effective and safe manner.

Decreased nutrient absorption, reduced appetite, and increased thermogenesis are being considered as possible pharmacological treatments for obesity. All of them have their own significant drawbacks. Decreased nutrient absorption (including, for example, impaired fat absorption) can cause gastrointestinal discomfort. Usually,

- 1 040171 Appetite reduction is expected to involve effects on brain structures, thereby leading to brain targeting problems and contamination of other tissues. Increasing thermogenesis can also have serious side effects.

Obesity treatment.

For effective treatment of obesity, current treatments typically include diet and exercise programs, lifestyle management, pharmacotherapy, and surgery. Treatment decisions are generally made based on the severity of obesity, the severity of underlying medical conditions, the subject's risk category, and the subject's expectations.

Marked improvements in cardiovascular risk and the occurrence of diabetes have been observed, along with a loss of 5-10% of total weight, following clinical guidelines for the treatment of obesity, which recommend a target cut-off of 10% body weight loss from baseline. Unfortunately, available drug therapies to promote weight loss do not provide adequate benefit to most obese subjects due to side effects, contraindications, or lack of response (National Heart, Lung and Blood Institute, Clinical guidelines on the identification, evaluation, and treatment of overweight and obesity in adults: the evidence report, NIH Publication No. 98-4083, September 1998).

Treatment of obesity with drugs (means).

Current treatment options for type IIb hyperlipidemia, which occurs in most obese people, are limited. Although statins are very effective in lowering LDL-C, they are generally not very effective in lowering triglyceride levels. Some statins at high dose levels, such as atorvastatin 80 mg, significantly lower triglyceride levels. However, treatment with high doses of statins can cause muscle pain (myalgia) and is often not well tolerated by patients. In addition, treatment with high doses of statins carries with it an increased risk of serious muscle toxicity such as rhabdomyolysis. Another treatment, which typically involves the administration of a combination of a cholesterol-lowering agent, such as a statin, and a triglyceride-lowering agent, such as fibrate, niacin, or fish oil. However, commonly used triglyceride-lowering agents may not be suitable or may not be well tolerated, for example, fibrates are associated with myalgia and an increased risk of muscle toxicity, fish oil must be taken multiple times daily, and niacin causes hot flushes, especially when given in combination with statins.

The most common over-the-counter drugs for the treatment of obesity are phenylpropanolamine and ephedrine, while the most common prescription drug is fenfluramine. These medicines have been removed from the market for safety reasons. Drugs approved for the long-term treatment of obesity fall into two categories.

1. Amphetamines and sibutramine, which act on the hypothalamus to control appetite stimulation in the CNS. (Appetite suppressants via the CNS).

2. Orlistat, which is a lipase inhibitor that blocks fat absorption in the gastrointestinal tract and reduces energy absorption (Gut Lipase Inhibitors [Cooke et al 2006 Nat Rev Drug Discov 5:919-31].).

Common side effects associated with these drugs include tachycardia, hypertension, encopresis, and/or valvular failure, making the development of anti-obesity drugs of paramount importance.

Although appetite suppressants and lipase inhibitors function in the gut through very different mechanisms, they share the same goal of reducing body weight while reducing the amount of calories that enter the general circulation. Unfortunately, these indirect treatments result in only small initial weight loss (approximately 5% compared to placebo), which is usually not maintained. After one or two years of treatment, most patients regain or exceed their original weight. In addition, most approved anti-obesity therapies have unwanted and often dangerous side effects that can complicate treatment and interfere with the patient's quality of life.

For example, orlistat, sibutramine and liraglutide, i.e. the components used until now to treat metabolic syndrome or obesity cause a number of serious side effects. In particular, the European Medicines Agency (EMA) reviewed the safety and efficacy of sibutramine and concluded that the benefits of sibutramine do not outweigh its risks, and that all marketing authorizations for medicinal products containing sibutramine should temporarily cease to be valid in the territory. throughout Europe (see Questions and answers on the suspension of medicines containing sibutramine - Outcome of a procedure under Article 107 of Directive 2001/83/EC, EMA/808179/2009). In addition, the main side effects of orlistat include steatorrhea, encopresis, and inhibition of the absorption of fat-soluble vitamins, thereby causing the patient taking orlistat to:

- 2 040171 (i) avoid high-fat foods and (ii) separately take dietary supplements containing fat-soluble vitamins and other fat-soluble nutrients.

Treatment of obesity through surgery.

Bariatric surgery can be considered a weight loss intervention for subjects whose BMI is 40 kg/m ² or greater. Subjects with a BMI of 35 kg/m ² and an associated serious medical condition are also potential subjects for this treatment option. Unfortunately, postoperative complications often result from bariatric surgical procedures, including hemorrhage, embolism or thrombosis, wound complications, deep suppuration, respiratory complications, and gastrointestinal blockages; during the postoperative period, a second operation is sometimes required to eliminate these complications. Significant and serious adverse events associated with bariatric surgery are common, occurring in approximately 4 percent of procedures performed (including death in 0.32% of subjects undergoing laparoscopic banding or bypass, respectively).

Lifestyle.

The most important advice given to people with this condition is aerobic exercise, regulation of diet and eating behavior, and weight loss (whether obese or overweight). Since the effectiveness of lifestyle changes has not been confirmed, therapeutic strategies are required. However, potent and safe therapeutic strategies for these diseases are imperfect. There are currently no effective pharmaceutical treatments to address this pandemic problem. Although surgical procedures can reduce weight by 50-90%, this is limited by the risk of surgery and associated side effects. Thus, there is still a need to develop alternative and/or improved therapeutic strategies for the treatment of obesity and its associated disorders.

Medical treatment of NAFLD.

Treatment of symptoms of NAFLD/NASH includes lipid-lowering agents, insulin sensitizers (agents), and reduction of inflammation through antioxidant agents (such as vitamin E, selenium, and betaine), anti-apoptotic agents, and anti-cytokine agents, and may also include reduction of total cholesterol, loss weight control, control of any associated diabetes, reduction or elimination of alcohol consumption, treatment of high blood pressure, and regular exercise. (American College of Gastroenterology; WebMD).

The most important advice given to people with this condition is to reduce their weight (if they are obese or overweight), eat a balanced and healthy diet, increase physical activity, abstain from alcohol, and avoid unnecessary medications. Another experimental approach for the treatment of NAFLD/NASH is the use of newer antidiabetic drugs, even for people without diabetes. The main goal of clinical treatment of NAFLD/NASH is to reduce the risk of cardiovascular disease and type II diabetes. The risks of these diseases are greatly reduced by lowering blood triglyceride levels, including LDL cholesterol, lowering blood pressure and lowering blood glucose levels.

Current treatments target insulin resistance (eg, metmorphine, thiazolidinediones) or insulin release from beta cells (eg, sulfonylurea, exenatide). However, these treatments have various disadvantages, including side effects, limited efficacy, and undesirable long-term effects. However, there are currently no approved treatments for NASH/NAFLD per se.

NAFLD can continue and cause cirrhosis or liver cancer, so early and effective treatment is important. Otherwise, the only treatment option for patients with advanced liver cirrhosis is liver transplantation. However, this procedure can be performed in a small number of patients due to the presence of surgical contraindications and organ deficiency.

Although there have been continued advances, there remains a strong need for safer, more effective and improved therapies for the treatment of obesity and its associated diseases such as NASH/NAFLD. Thus, compound(s) of natural origin, which is(are) active(s) when administered orally, which(s) will reduce obesity and related diseases, are desirable.

The need for natural active ingredients.

OEA is an endogenous peroxisome proliferator-activated receptor alpha (PPAR-α) agonist and is the result of a combination of oleic acid and ethanolamine. OEA is a naturally occurring lipid mediator that inhibits food intake and body weight gain and has therefore been a molecule of recent scientific interest in the search for therapeutic strategies for the treatment of obesity in humans. OEA is a fatty acid metabolite that is produced by the gut to signal satiety to the brain, thereby regulating appetite and healthy body fat. OEA contributes to the regulation of lipid metabolism, promotes hunger control by sending appetite-suppressing messages to the brain, and has also been shown to help lower blood levels of both triglyceride and cholesterol.

P-aminoisobutanoic acid (BAIBA) is a non-protein p-amino acid, a thymine catabolite, which is further broken down into propionyl-CoA, methylmalonyl-CoA, and succinyl-CoA within the mitochondria, especially in the liver. BAIBA can also be produced by catabolism of the branched chain amino acid valine. BAIBA reduces body fat percentage through increased fatty acid oxidation (FAO) and reduced primary liver lipogenesis in mice. BAIBA enhances brown staining of white adipose tissue and FFA in the liver via peroxisome proliferator-activated receptor α (PPARα) and may contribute to protection against exercise-induced metabolic disorders. In humans, BAIBA concentrations increase with exercise and are inversely associated with metabolic risk factors. Thus, BAIBA may contribute to protection against exercise-induced metabolic disorders.

Antioxidants also play an important role in the treatment of obesity. An increase in leptin leads to an increase in adhesion molecules; adipocytes attract monocytes, which then transform into macrophages. Macrophages are capable of producing large amounts of oxidants such as peroxynitrites. Obesity-associated increase in oxidative stress has been investigated by various methods of measuring thiobarbituric acid reactive substances (TBBA), malondialdehyde (MMD) production, oxidized LDL, oxidized urinary albumin, peroxide assessed by reactive oxygen assays, and protein carbonyl formation in erythrocytes. A review of the literature indicates that, in most studies, obesity and type 2 diabetes are associated with increased oxidative stress. Thus, oxidative stress is also associated with weight loss and not with diet per se. Given that obesity and type 2 diabetes are associated with increased oxidative stress, there is a need for an antioxidant to control excess free radicals or toxins produced.

α-glucosidase inhibitors can be used as a new class of antidiabetic drug. By competitive inhibition of glucosidase activity, these inhibitors help prevent the rapid breakdown of sugars and thereby control blood sugar levels. In the 1980s, α-glucosidase inhibitors became a new class of antidiabetic drug. Inhibitors of α-glucosidase slow down the process of assimilation and absorption of carbohydrates by competitively blocking the activity of glucosidase. All of them contain sugar components and their synthesis involves cumbersome multi-stage procedures. Moreover, clinically synthesized α-glucosidase inhibitors have been associated with severe gastrointestinal side effects. Thus, there is a need to find alternatives that can display α-glucosidase inhibitory activity without side reactions.

1-deoxynojirimycin (DNJ) is a glucose analog with a minor amino group instead of an oxygen atom in the pyranose ring of glucose. DNG potently inhibits α-glucosidase in the small intestine by binding to the active site of α-glucosidase. DNJ has also been found in the leaves and fruits of M. alba, suggesting that the well-known effect of mulberry leaves may be due to DNJ inhibiting postprandial hyperglycemia by inhibiting α-glucosidase in the small intestine. The use of DNG-enriched mulberry leaf extract slightly reduced triglyceride levels after long-term use (12 weeks), although the reduction was not statistically significant. Several studies have demonstrated that phytochemicals from natural resources provide new options for the treatment of diabetes. Mulberry leaf extracts have been used in China and other Asian countries to treat diabetes based on reports of anti-diabetic effects in experimental animals.

Cannabinoid-based drugs have therapeutic potential for the treatment of pain. Increasing endocannabinoid levels with inhibitors of fatty acid amide hydrolase (FAAH) has an analgesic effect. Fatty acid amide hydrolase (FAAH) is a membrane-bound serine hydrolase that belongs to the motif of the amidase hydrolase family. The FAAH enzyme breaks down fatty acid amides such as anandamide (N-arachidonoylethanolamine), noleoylethanolamide (N-OEA), N-palmitoyletanolamide (N-PEA), and oleamide. FAAH belongs to a broad and different class of enzymes called the amidase (AS) family motif. FAAH inhibitors are a class of molecules that deactivate fatty acid amide hydrolase enzymes by preventing the hydrolysis of anandamide, oleoylethanolamide and palmitoylethanolamide, thereby increasing their endogenous levels. Known chemically synthesized FAAH inhibitors are BIA 10-2474, URB524, URB-597, URB694, URB937, etc. These inhibitors are disclosed in a published article by Colombano et al. called O-(Triazolyl) methyl carbamates as a novel and potent class of

- 4 040171 fatty acid amide hydrolase (FAAH) inhibitors and in an article published by Otrubova et al. Entitled

The discovery and development of inhibitors of fatty acid amide hydrolase (FAAH).

Related prior art

WO02080860 relates to a method for reducing food intake in mammals by using

OEA.

WO200434968 relates to a method for reducing food intake in mammals by administering a PPARa agonist and a chemically synthesized CB-1 receptor agonist.

WO2004045307 describes food products and supplements containing OEA for satiety induction and weight loss.

WO2004078113 relates to a method for preventing or treating obesity and obesity-related complications by administering a Cox-2 inhibitor in combination with one or more weight-reducing agents selected from the group consisting of OEA.

WO200546580 describes the use of OEA to reduce body weight, modulate lipid metabolism in the body and reduce food intake.

WO2007006319 discloses the use of an appetite suppressant or satiety inducing agent, such as OEA, for the manufacture of a pharmaceutical composition for the prevention or therapeutic treatment of diseases or disorders associated with impaired appetite regulation in mammals.

WO2009142713 relates to a method for reducing lipid absorption in an animal, comprising administering to the animal one or more fatty acid alkanolamides, such as OEA, in an amount effective to reduce lipid absorption by the animal.

WO201290225 discloses a fixed dose pharmaceutical composition containing co-crystals of metmorphine or pharmaceutically acceptable salts thereof, as well as OEA with other antidiabetic agents.

WO2012154711 discloses a composition containing OEA in a therapeutically effective amount for accelerating the growth of lean body mass.

CN102579414 relates to the use of OEA as an appetite suppressant.

WO201437546 discloses a fluid emulsion containing oleoylethanolamide, oil, water and an emulsifier, as well as a free-flowing powder containing oleoylethanolamide.

WO2016185468 discloses a pharmaceutical composition comprising a therapeutically effective amount of a mixture of at least one opioid or salt thereof and at least one Nacylethanolamine or salt thereof, wherein the molar ratio between the opioid or salt thereof and Nacylethanolamine or salt thereof is from about 1:1 to about 1:100.

WO201725588 describes a nanoparticle containing at least one of a biocompatible lipid and a lipophilic therapeutic agent such as oleoylethanolamide, a lubricant and/or a diagnostic agent, the nanoparticle having a surface containing a water soluble polymer.

There are several patent applications (WO 2012/015704, WO 2013/028570, WO 2014/017936, WO 2015/07613, WO 2015/07615, WO 2015/012708, WO 2015/016728, WO 2015/157313), in which disclosed chemically synthesized various types of FAAH inhibitors that increase the level of OEA. However, these chemically synthesized FAAH inhibitors may have side effects when administered to humans or animals. Several studies have identified serious side effects (including death) of chemically synthesized FAAH inhibitors. Some of these include Eddleston Michael et al; Implications of the BIA-102474-101 study for review of firstinto-human clinical trials, Br J Clin Pharmacol (2016) 81 582-586; Mallet et al.; FAAH inhibitors in the limelight, but regrettably, International Journal of Clinical Pharmacology and Therapeutics, Vol. 54 - no. 7/2016(498-501); and Kaur et al. What failed BIA 10-2474 Phase I clinical trial? Global speculations and recommendations for future Phase I trials, J Pharmacol Pharmacotherapy. 2016 Jul-Sep; 7(3): 120-126.

In particular, there are many problems with the treatment of obesity and associated disorders associated with fat metabolism, such as NAFLD, NASH, such as those disclosed in the prior art documents above. In addition, as described above, currently available treatments for obesity and its associated disorders are ineffective and complicated due to relapse rates and a wide range of side effects such as endothelial dysfunction and cardiovascular risk. The inventors in the present application have identified the urgent need to provide effective and safe treatment of patients with obesity. Thus, the inventors have developed a pharmaceutical composition/formulation containing a synergistic combination of OEA with other active ingredients that control dietary obesity, as well as treat other metabolic disorders associated with fat metabolism, such as NASH, NAFLD, with an acceptable safety profile.

There is also a need for an effective composition/composition for the treatment of excess weight as well as for supporting weight loss.

Therefore, there is also an urgent need to develop treatments for delaying the development, prevention of formation or inversion of the fatty liver condition. Currently available treatments for NAFLD and NASH are quite limited and there is a need for safe and effective treatment of patients suffering from NAFLD and NASH.

Despite the attempts made in the prior art, the treatment of obesity and related disorders remains difficult. Thus, it would be beneficial to provide effective and safe treatment to prevent obesity-related complications.

Disclosure of the essence of the invention

The present invention provides a pharmaceutical composition for the treatment of obesity and related disorders, comprising a combination of:

42.55% by weight oleoylethanolamide (OEA),

2.13% by weight of valine and

31.91% by weight pantethine.

In a preferred aspect, the pharmaceutical composition of the present invention further comprises a natural alpha-glucosidase inhibitor, a fatty acid amide hydrolase (FAAH) inhibitor, or a naturally occurring natural alpha-glucosidase inhibitor, or a combination thereof.

In one embodiment, the natural alpha-glucosidase inhibitor in the pharmaceutical composition is 1-deoxynojirimycin.

The amount of natural alpha-glucosidase inhibitor may range from 0.5 to 1.5% by weight of the composition.

Further, the natural FAAH inhibitor in the pharmaceutical composition can be selected from quercetin, myricetin, isorhamnetin, kaempferol, pritimerin, curcumin, biochanin A, genistein and daidzein.

Preferably, the amount of natural FAAH inhibitor in the pharmaceutical composition is in the range of 0.5 to 25% by weight of the composition.

Additionally, the pharmaceutical composition of the invention contains pharmaceutically acceptable excipients.

Said pharmaceutically acceptable excipients may be selected from diluent, binder, lubricant, glidant and solvent.

The amount of said diluent may range from 1 to 30% by weight of the composition, the amount of binder may range from 0.5 to 1.5% by weight of the composition, the amount of lubricant may range from 0.5 to 5% by weight. by weight of the composition, the amount of glidant may be in the range of 1.0 to 25% by weight of the composition, or the solvent may be taken in sufficient quantity.

The pharmaceutical composition according to the invention may be in the form of a tablet, capsule, sachet, pill, liquid or solid filled hard capsule, soft capsule, powder, granule, suspension, solution or modified release formulation.

In another aspect of the present invention, a method for preparing a pharmaceutical composition of the invention is described. The method includes:

(a) separately weighing the OEA, valine and pantethine in separate containers, (b) sieving the pre-weighed OEA, valine and pantethine separately, (c) mixing the contents in step b, (d) preparing the binder solution in a separate container and adding it to the step c to obtain a paste, and sifting the paste to obtain granules, (e) drying the obtained granules to reduce the dryness level (DS) to less than 1.5% v / v, (f) sieving the dried granules through a suitable sieve and (g) screening the pre-weighed lubricant(s) or glidant(s) separately through a suitable sieve and adding to step f.

Brief description of the drawings

Fig. 1 - study comparing the increase in percentage of body weight in different groups of animals and the effect of the introduction of the study composition/formulation.

Fig. 2 is a study comparing percentage reduction in serum triglyceride levels in various groups of animals and the effect of administering the test composition/formulation.

Fig. 3 is a study comparing the percentage reduction in total serum cholesterol in various groups of animals and the effect of administration of the study composition/formulation.

Fig. 4 is a study comparing percentage reduction in serum LDL levels in different groups of animals and the effect of administration of the study composition/formulation.

Fig. 5 is a study comparing the percentage reduction in hepatic triglyceride levels in different groups of animals and the effect of administering the test composition/formulation.

Fig. 6 is a study comparing the reduction in percentage of total liver cholesterol in different groups of animals and the effect of administration of the study composition/formulation.

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Detailed description of the invention

While the invention has been described and illustrated with reference to certain embodiments, one skilled in the art will appreciate that various adaptations, changes, modifications, substitutions, deletions, or additions of procedures and protocols can be made without departing from the spirit and scope of the invention. For example, effective dosages other than the specific dosages set forth herein above may be employed as a result of variation in the response of the mammal being treated to any indication of the compounds of the invention as set forth above. The specific pharmacological responses observed may vary according to and depending on the specific active compounds selected along with the pharmaceutical carriers present. In addition, responses may vary depending on the type of formulation used and the mode of administration, such expected variations or differences in results are expected in accordance with the objectives and practical implementations of the present invention. Thus, it is intended that the invention be defined by the scope of the claims, and that such claims should be interpreted broadly as appropriate.

The present invention is directed to a pharmaceutical composition/composition containing a synergistic combination of OEA with other natural active ingredients that effectively reduces body weight. In addition, the pharmaceutical composition/composition improves liver function by significantly reducing the level of triglyceride and cholesterol in the liver, and also reduces steatosis, regardless of the etiology of the pathology. In addition, it may be administered to prevent NAFLD and NASH in a subject suffering from metabolic syndrome and/or obesity and/or diabetes.

The inventors have carried out exhaustive scientific studies along with pre-clinical studies and have found that the combination of OEA with other natural ingredients of the present invention is extremely effective in the treatment of obesity and associated fat metabolism disorder such as NAFLD and NASH by significantly lower serum and liver triglyceride and cholesterol levels.

Within the scope of the present invention, it has been found that the synergistic combination of OEA with other natural active ingredients has unexpected and particularly advantageous properties. This makes them particularly useful in the treatment and/or prevention (including preventing, slowing, delaying and/or reversing progression or reducing the incidence or delaying outcome) of NAFLD, including hepatic steatosis, NASH and/or hepatic fibrosis, and/ or disorders associated with them, as well as to prevent cirrhosis of the liver (irreversible ongoing scarring of the liver) and / or hepatocellular carcinoma.

The aim of the present invention is to provide a pharmaceutical composition/composition containing a synergistic combination of OEA with other natural active ingredients that effectively reduces body weight.

In an embodiment, the present invention provides a pharmaceutical composition/formulation comprising a synergistic combination of OEA with other natural active ingredients for use in the treatment and/or prevention of obesity and associated fat metabolism disorder such as NASH, NAFLD.

The present invention also provides a pharmaceutical composition/formulation containing a synergistic combination of OEA with other natural active ingredients for reducing serum and liver triglyceride and cholesterol levels.

The present invention further provides a pharmaceutical composition/formulation comprising a synergistic combination of OEA with other natural active ingredients for the treatment, delay, progression and/or prevention of fatty liver diseases resulting from obesity, including, but not limited to, alcoholic fatty liver simplex, alcoholic steatohepatitis (ASH), alcoholic liver fibrosis, alcoholic cirrhosis, NAFLD, non-alcoholic simple fatty liver, NASH, non-alcoholic liver fibrosis and non-alcoholic cirrhosis.

The composition/formulation of the present invention may also reduce body weight and alter fat metabolism.

Another object of the present invention is to provide a composition/formulation containing a synergistic combination of OEA with natural active ingredients. Natural active ingredients can be selected from natural BAIBA, natural FAAH inhibitor, natural alpha-glucosidase inhibitor, natural antioxidant, or combinations thereof.

In an aspect, the present invention provides a composition/formulation for the treatment of obesity and a related disorder associated with fat metabolism, said composition/formulation comprising a synergistic combination of:

(a) oleoylethanolamide (OEA);

(b) natural β-aminoisobutanoic acid (BAIBA);

(c) a natural antioxidant; And

- 7 040171 (d) optionally, one or more other natural active ingredients selected from:

(i) a natural fatty acid amide hydrolase (FAAH) inhibitor, (ii) a natural alpha-glucosidase inhibitor.

Therefore, the main object of the present invention is to provide a pharmaceutical composition/composition containing a synergistic combination of OEA with natural BAIBA and/or natural antioxidant. In one aspect, the composition/formulation may optionally comprise a natural alpha-glucosidase inhibitor and/or a naturally occurring FAAH inhibitor.

It is known in the art that OEA plays an important determining role in lipid metabolism and can enhance fatty acid oxidation in primary cultured skeletal muscle cells, liver cells and cardiomyocytes. An animal study confirmed that OEA regulates fatty acid metabolism by activating PPAR-α. Obesity and insulin resistance are often accompanied by lipid metabolism disorders, which are the main cause of NAFLD. OEA, an endogenous PPAR-α agonist, has a beneficial effect on high-fat diet-induced NAFLD in rats by promoting fatty acid β-oxidation and re-inhibiting lipogenesis. In the prior art, OEA has been considered as an improved regulator for the treatment of obesity, atherosclerosis, and other metabolic disorders. However, the effects of the composition/formulation of the present invention, containing a synergistic combination of OEA with other natural ingredients, to improve the treatment of alimentary obesity and fatty liver diseases resulting from obesity, have not been recorded. This synergistic combination of the present invention plays a fundamental role in the treatment of obesity, which is often accompanied by lipid disorders and is the main cause of NAFLD. The combination of OEA with other natural active ingredients is capable of providing a safe pharmaceutical formulation with enhanced or synergistic effects compared to OEA alone in the treatment of dietary obesity and fatty liver disease resulting from obesity.

The composition/composition of the present invention containing OEA, natural BAIBA, a natural antioxidant, together with other natural ingredients such as an alpha glucosidase inhibitor, is helpful in the treatment of obese/obese people suffering from diabetes.

Side effects from the use of synthetic ingredients (such as a chemically synthesized FAAH inhibitor) in pharmaceutical compositions are widely known. The composition/composition of the present invention contains natural FAAH inhibitors that inhibit the breakdown of OEA and increase the level of OEA in the body without any side effects.

In a preferred aspect, the ratio of OEA:natural BAIBA:natural antioxidant is in the range of 15-55:2-61:23-41. In yet another preferred aspect, the ratio of OEA:natural BAIBA:natural antioxidant is 55:2:41.

The pharmaceutical composition/composition of the present invention contains OEA, and the amount of OEA in the pharmaceutical composition/composition of the present invention is in the range from 10 to 70% by weight of the composition/composition. In an embodiment, the amount of OEA is in the range of 20 to 70% by weight. In still another embodiment, the amount of OEA is in the range of 30 to 70% by weight. In still another embodiment, the amount of OEA is in the range of 40 to 70% by weight. In still another embodiment, the amount of OEA is in the range of 50 to 70% by weight. In still another embodiment, the amount of OEA is in the range of 60 to 70% by weight.

In a preferred embodiment, the implementation of the pharmaceutical composition/formulation of the present invention contains OEA, and the amount of OEA in the pharmaceutical composition/formulation is in the range from 100 to 1000 mg per unit dose.

The pharmaceutical composition/formulation of the present invention contains a natural BAIBA selected from amino acids such as valine, isoleucine or the like. The amount of natural BAIBA in the pharmaceutical composition/formulation of the present invention ranges from 2 to 60% by weight of the composition/formulation. In an embodiment, the amount of natural BAIBA is in the range of 5 to 60% by weight. In still another embodiment, the amount of natural BAIBA is in the range of 10 to 60% by weight. In still another embodiment, the amount of natural BAIBA is in the range of 20 to 60% by weight. In yet another embodiment, the amount of natural BAIBA is in the range of 30 to 60% by weight. In still another embodiment, the amount of natural BAIBA is in the range of 40 to 60% by weight. In yet another embodiment, the amount of natural BAIBA is in the range of 50 to 60% by weight.

In a preferred embodiment, the pharmaceutical composition/formulation of the present invention contains natural BAIBA selected from valine, isoleucine, or the like, wherein the amount of natural BAIBA in the pharmaceutical composition/formulation is in the range of 10 mg to 3000 mg per unit dose.

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The pharmaceutical composition/composition of the present invention contains a natural antioxidant selected from vitamin E, glutathione, selenium, pantethine, fucoxanthin or the like. The amount of natural antioxidant in the pharmaceutical composition/composition of the present invention is in the range of 20 to 35% by weight of the composition/composition. In an embodiment, the amount of natural antioxidant is in the range of 25 to 35% by weight. In another embodiment, the amount of natural antioxidant is in the range of 30 to 35% by weight.

In a preferred embodiment, the pharmaceutical composition/formulation of the present invention contains a natural antioxidant selected from selenium, glutathione, vitamin E, pantethine, fucoxanthin, or the like, wherein the amount of the natural antioxidant in the pharmaceutical composition/formulation is in the range of 0.1 to 500 mg for the standard dose.

In yet another aspect, the pharmaceutical composition/formulation of the present invention contains a synergistic combination of OEA, valine, pantethine and pharmaceutically acceptable excipients.

The pharmaceutical composition/composition of the present invention optionally contains other ingredients selected from a naturally occurring alpha-glucosidase inhibitor or a naturally occurring FAAH inhibitor, or a combination thereof.

The pharmaceutical composition/formulation of the present invention further comprises a natural alpha-glucosidase inhibitor, such as 1-deoxynojirimycin, which is derived from mulberry leaves. The amount of natural alpha-glucosidase inhibitor in the pharmaceutical composition/composition of the present invention is in the range of 0.5 to 1.5% by weight of the composition/composition.

In a preferred embodiment, the pharmaceutical composition/composition of the present invention comprises a natural alpha-glucosidase inhibitor such as 1-deoxynojirimycin, wherein the amount of the natural alpha-glucosidase inhibitor in the pharmaceutical composition/formulation is in the range of 1 to 40 mg per unit dose.

The pharmaceutical composition/composition of the present invention contains natural FAAH inhibitors selected from quercetin, myricetin, isorhamnetin, kaempferol, pritimerin, curcumin, biochanin A, genistein, daidzein, or the like. The amount of natural FAAH inhibitor in the pharmaceutical composition/formulation of the present invention is in the range of 0.5 to 25% by weight of the composition/formulation.

In a preferred embodiment, the pharmaceutical composition/formulation of the present invention contains a natural FAAH inhibitor selected from quercetin, myricetin, isorhamnetin, kaempferol, pristimerin, curcumin, biochanin A, genistein, daidzein, or the like, wherein the amount of the natural FAAH inhibitor in the pharmaceutical composition/formulation is is in the range of 1 mg to 100 mg per standard dose.

The pharmaceutical composition/composition of the present invention may be in the form of tablets, capsules, granules, powder, sachets, suspension, solution, modified release formulations, topical formulations, and the like. The compositions of the present invention contain suitable excipients such as diluents, disintegrants, binders, solubilizing agents, lubricants, glidants, solvents, etc.

In a preferred embodiment, the implementation of the pharmaceutical composition/formulation of the present invention may be made for oral administration. For oral administration, a solid pharmaceutical composition may be in the form of, but not limited to, tablets, capsules, pills, hard capsules filled with liquids or solids, soft capsules, powders or granules, sachets, and the like. The compositions may additionally contain pharmaceutically acceptable excipients. Preferred excipients are selected from diluent, disintegrant, binder, glidant, etc.

Diluents are selected from microcrystalline cellulose, lactose (anhydrous/monohydrate/spray dried), starch, cellulose powder, silicified microcrystalline cellulose, ammonium alginate, calcium carbonate, calcium lactate, dibasic calcium phosphate (anhydrous/dibasic dihydrate/tribasic), calcium silicate, sulfate calcium, cellulose acetate, pressed sugar, confectionery sugar, corn starch, pregelatinized starch, dextrates, dextrin, dextrose, erythritol, ethyl cellulose, fructose, fumaric acid, glyceryl palmitostearate, isomalt, kaolin, lactitol, magnesium carbonate, magnesium oxide, maltodextrin, maltose , mannitol, medium chain triglycerides, polydextrose, polymethacrylates, simethicone, sodium alginate, sodium chloride, sorbitol, sterilizable maize, sucrose, sugar spheres, β-cyclodextrin sulfobutyl ether, talc, tragacanth, trehalose, xylitol, or the like. The amount of diluent in the pharmaceutical composition/composition of the present invention ranges from 1 to 30% by weight of the composition/composition.

The binder is selected from hypromellose, starch, gum arabic, agar, alginic acid, calcium carbonate, calcium lactate, carbomers, sodium carboxymethyl cellulose, carrageenan, cellulose acetate phthalate, carob, chitosan, copovidone, corn starch, pregelatinizi

- 9 040171 starch, cottonseed oil, dextrates, dextrin, dextrose, ethylcellulose, gelatin, glycerol behenate, guar gum, hydrogenated vegetable oil type I, hydroxyethylcellulose, hydroxyethylmethylcellulose, hydroxypropylcellulose, inulin, lactose, liquid glucose, low-substituted hypromellose, magnesium aluminosilicate, maltodextrin, maltose, methylcellulose, microcrystalline cellulose, pectin, poloxamer, polycarbophil, polydextrose, polyethylene oxide, polymethacrylates, povidone, sodium alginate, stearic acid, sucrose, sunflower oil, tricaprylin, vitamin E polyethylene glycol succinate, zein or the like. The amount of binder in the pharmaceutical composition/composition of the present invention is in the range of 0.5 to 1.5% by weight of the composition/composition.

The lubricant is selected from magnesium stearate, zinc stearate, calcium stearate, glycerol monostearate, glycerol behenate, glycerol palmitostearate, hydrogenated castor oil, hydrogenated type I vegetable oil, light mineral oil, magnesium lauryl sulfate, medium chain triglycerides, mineral oil, myristic acid, palmitic acid , poloxamer, polyethylene glycol, sodium benzoate, sodium chloride, sodium lauryl sulfate, sodium stearyl fumarate, stearic acid, talc, potassium benzoate, or the like. The amount of lubricant in the pharmaceutical composition/composition of the present invention is in the range of 0.5 to 5% by weight of the composition/composition.

The glidant is selected from colloidal silicon dioxide, talc, tribasic calcium phosphate, powdered cellulose, hydrophobic colloidal silicon dioxide, magnesium oxide, magnesium silicate, magnesium trisilicate, silicon dioxide, or the like. The amount of glidant in the pharmaceutical composition/composition of the present invention ranges from 1 to 25% by weight of the composition/composition.

The solvent is selected from water, alcohol, isopropyl alcohol, propylene glycol, almond oil, benzyl alcohol, benzyl benzoate, butylene glycol, silicon dioxide, castor oil, corn (maize) oil, cottonseed oil, dibutyl phthalate, diethyl phthalate, dimethyl ether, albumin, dimethyl phthalate, dimethyl sulfoxide, dimethylacetamide, ethyl acetate, ethyl lactate, ethyl oleate, glycerin, glycofurol, isopropyl myristate, isopropyl palmitate, light mineral oil, medium chain triglycerides, methyl lactate, mineral oil, monoethanolamine, octyldodecanol, olive oil, walnut oil, polyethylene glycol, polyoxyl 35 castor oil, propylene carbonate, pyrrolidone, safflower oil, sesame oil, soybean oil, sunflower oil, triacetin, tricaprylin, triethanolamine, triethyl citrate, triolein, water-soluble solvents or the like. The solvent in the pharmaceutical composition/composition of the present invention is used in an amount from sufficient to 100% by weight of the composition/composition.

The development of pharmaceutical compositions or formulations where one or more ingredients are derived from natural sources, such as BAIBA, an antioxidant, an alpha-glucosidase inhibitor, and an FAAH inhibitor, is problematic for the developer. Such problems are associated with providing a dosage form of a suitable size, while containing an effective amount of active ingredients. Problems are also associated with providing stable formulations while maintaining the desired pharmacokinetic properties. According to publicly available information, the combination of OEA with natural BAIBA and a natural antioxidant and/or a natural alpha-glucosidase inhibitor and/or a natural FAAH inhibitor is currently unknown. The present invention provides a stable and therapeutically effective composition/composition containing OEA, natural BAIBA, natural antioxidant and optionally containing natural alpha-glucosidase inhibitor, natural FAAH inhibitor.

Some preferred compositions/formulations of the present invention are as follows:

Composition/composition 1

Composition/composition 2

- 10 040171

Composition/composition 3

No. n/n Ingredient Quantity (% by weight) 1 . OEA 10-70% 2. Natural BAIBA 2-60% 3. natural antioxidant 20-35% 4 . natural inhibitor of alpha glucosidases 0.5-1.5%

Composition/composition 4

No. p / p Ingredient Quantity (% by weight) 1 . OEA 10-70% 2. Natural BAIBA 2-60% 3. natural antioxidant 20-35% 4 . Natural FAAH Inhibitor 0.5-25% 5. natural inhibitor of alpha glucosidases 0.5-1.5%

General method for obtaining the composition/composition of the present invention.

1) Accurate weighing of all materials in separate containers.

2) Screen the pre-weighed OEA, natural antioxidant, natural BAIBA, diluent(s) and glidant(s) through a #40 sieve.

3) Mixing the contents from step 2 in a high speed mixer granulator with agitator at low speed.

4) Obtaining a binder solution: Weighing the binder(s) means(s) and its(their) dissolution in the solvent(s) in a separate container to obtain a clear transparent solution.

5) Adding binder solution to step 3 in WASH at low stirrer speed.

6) Screening and drying the resulting granulated wet mass in a fluid bed dryer at 50+5° C. until the dryness level (DS) of the mixture is reduced to less than 1.5% v/v.

7) Screen the dried granules through a #18 sieve and a #24 sieve.

8) Screen the pre-weighed lubricant(s) or glidant(s) separately through a #40 sieve and blend with step 7.

9) Filling the mixture into HPMC capsule shells and curing.

10) Transferring the filled capsules to a polishing dispenser and a visual inspection machine to remove powder residue adhering to the capsule shells.

Examples.

It should be understood that the following examples are given only to illustrate the present invention. Some modifications to the products and/or methods used may be implemented and still achieve the objectives of the invention. It is assumed that such modifications are included in the scope of the claimed invention. For experiments, OEA and natural antioxidant were obtained from Wuxi Cima Science Co., Ltd. No.288 Shibawan Road Wuxi 214064 Jiangsu, China; Natural BAIBA was obtained from Shantou Jiahe Biologic Technology Co., Ltd. Address: No.4 Lane 1 Xigang Road, Shantou, 515021, P.R. China; 1-deoxynojirimycin (DNJ) derived from mulberry leaves was obtained from Bio-gen Extracts Pvt. Ltd, No. 39/2, 3rd Floor, 8th Mile, T. Dasarahalli, Tumkur Road, Bengaluru- 560057, and natural FAAH inhibitors were obtained from Navpad Trade Impex, Office No.215, Sai Chamber, Near Bus Depot, Santacruz (East), Mumbai - 400055.

- 11 040171

Example 1

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide (OEA) 44.44 2 L-valine 2.22 3 Microcrystalline cellulose pH 101 29, 44 4 colloidal silicon dioxide 17.78 binder solution 5 HPMC 5 cps 0.56 6 Isopropyl alcohol in sufficient quantity 7 Methylene dichloride in sufficient quantity Extragranular Ingredients 8 colloidal silicon dioxide 2.78 9 magnesium stearate 2.78 General content 100.00

Example 2

No. p / p Ingredients % by weight Intragranular Ingredients 1 2 3 4 Solution ci 5 6 7 Oleoylethanolamide Isoleucine Dicalcium phosphate colloidal silicon dioxide stinging PVP-K 30 Isopropyl alcohol Methylene dichloride 40.00 45.00 8.50 4.00 0.50 in sufficient quantity in sufficient quantity Extragranular Ingredients 8 colloidal silicon dioxide 1.00 9 Talc 1.00 General content 100.00

Example 3

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 43.48 2 Fucoxanthin 21.74 3 Isoleucine 10, 87 4 Dicalcium phosphate 9, 24 5 colloidal silicon dioxide 8.70 binder solution 5 PVP-K 30 0.54 6 Isopropyl alcohol in sufficient quantity 7 Methylene dichloride in sufficient quantity Extragranular Ingredients 8 colloidal silicon dioxide 2.72 9 magnesium stearate 2.72 General content 100.00

- 12 040171

Example 4

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 50.00 2 L-valine 2.50 3 Daidzein 12.50 4 Anhydrous lactose 18.13 5 colloidal silicon dioxide 10.00 binder solution 6 Ethylcellulose 0.63 7 Isopropyl alcohol in sufficient quantity 8 Methylene dichloride in sufficient quantity Extragranular Ingredients 9 zinc stearate 3, 13 10 Talc 3, 13 General content 100.00

Example 5

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 57.14 2 L-valine 2.86 3 Genistein 1.14 4 Mannitol 20.29 5 colloidal silicon dioxide 11.43 binder solution 6 Starch 1.43 7 Isopropyl alcohol in sufficient quantity 8 Purified water in sufficient quantity Extragranular Ingredients 9 magnesium stearate 2.86 10 colloidal silicon dioxide 2.86 General content 100.00

Example 6

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 50.00 2 L-valine 2.50 3 Mericetin 25.00 4 Microcrystalline cellulose 101 pH 6, 25 5 colloidal silicon dioxide 10.00 binder solution 6 HPMC 5 cps 1.25 7 Isopropyl alcohol in sufficient quantity 8 Methylene dichloride in sufficient quantity Extragranular Ingredients 9 Talc 2.50 10 colloidal silicon dioxide 2.50 General content 100.00

- 13 040171

Example 7

Example 8

Example 9

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 57.14 2 L-valine 2, 86 3 Lactose monohydrate 14.29 4 colloidal silicon dioxide 17.14 binder solution 5 PVP K-30 1.43 6 DNJ (mulberry leaf extract) 1, 14 7 Isopropyl alcohol in sufficient quantity 8 Methylene dichloride in sufficient quantity Extragranular Ingredients 9 zinc stearate 3.00 10 Total sod< № p / p Intragra 1 2 3 4 5 Solution with 6 7 8 9 Vnegranulul 10 eleven Total sod № p / p Intragra 1 2 3 4 5 Talc erzhanie Ingredients zero ingredients Oleoylethanolamide L-valine Daidzein Lactose monohydrate colloidal silicon dioxide binder PVP k-30 DNJ (mulberry leaf extract) Isopropyl alcohol Methylene dichloride bright ingredients zinc stearate Talc holding | Ingredients zero ingredients Oleoylethanolamide L-valine Mericetin Dicalcium phosphate colloidal silicon dioxide 3.00 100.00 % by weight 50.00 2.50 12.50 12.50 15.00 1.25 100 in sufficient quantity in sufficient quantity 2, 63 2, 63 100.00 % by weight 50.00 2.50 25.00 7.75 10.00 binder solution 6 HPMC 5 cps 1.25 7 DNJ (mulberry leaf extract) 100 8 Isopropyl alcohol in sufficient quantity

- 14 040171

Example 10

Example 11.

No. p / p Ingredients % by weight 9 Methylene dichloride in sufficient quantity Extragranular Ingredients 10 magnesium stearate 1.25 eleven colloidal silicon dioxide 1.25 General content 100.00

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 40.00 2 Valine 2.00 3 Daidzein 10.00 4 Pantethine (dissolved) 30.00 5 Dicalcium phosphate 7.00 6 colloidal silicon dioxide 8.00 binder solution 7 HPMC 5 cps 1.00 8 Isopropyl alcohol in sufficient quantity 9 Methylene dichloride in sufficient quantity Extragranular Ingredients 10 magnesium stearate 1.00 eleven colloidal silicon dioxide 1.00 General content 100.00

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 44.44 2 L-valine 2.22 3 Genistein 0.89 4 Pantethine (dissolved) 33.33 5 Dicalcium phosphate 6, 89 No. p / p Ingredients % by weight 6 colloidal silicon dioxide 8.89 binder solution 7 PVP K-30 1, 11 8 Isopropyl alcohol in sufficient quantity 9 Methylene dichloride in sufficient quantity Extragranular Ingredients 10 magnesium stearate 1, 11 eleven colloidal silicon dioxide 1.11 General content 100.00

- 15 040171

Example 12.

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 44.44 2 L-valine 2.22 3 Pantethine (dissolved) 33.33 4 Dicalcium phosphate 6, 89 5 colloidal silicon dioxide 8.89 binder solution 6 HPMC 5 cps 1.11 7 DNJ (mulberry leaf extract) 0.89 8 Isopropyl alcohol in sufficient quantity 9 Methylene dichloride in sufficient quantity Extragranular Ingredients 10 magnesium stearate 1.11 eleven colloidal silicon dioxide 1, 11 General content 100.00

Example 13

№ p / p | Ingredients % by weight Intragame! No. p / p 1 2 3 4 5 6 Solution a 7 8 9 aular ingredients Ingredients Oleoylethanolamide L-valine Pantethine (dissolved) Daidzein Microcrystalline cellulose pH 101 Colloidal silica binder PVP k-30 DNJ (mulberry leaf extract) Isopropyl alcohol % by weight 40.00 2.00 30.00 10.00 6.20 8.00 100 0.80 in sufficient quantity 10 Methylene dichloride in sufficient quantity Extragranular Ingredients eleven zinc stearate 1.00 12 Talc 1.00 General content 100.00

- 16 040171

Example 14

Example 15

Example 16

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 44.44 2 L-valine 2.22 3 Pantethine (dissolved) 33, 33 4 Microcrystalline cellulose pH 101 6.00 5 colloidal silicon dioxide 8.89 binder solution 6 PVP K-30 1.11 7 DNJ (mulberry leaf extract) 0.89 8 Genistein 0.89 9 Isopropyl alcohol V enough

No. p / p Ingredients % by weight 10 Methylene dichloride in sufficient quantity Extragranular Ingredients eleven zinc stearate 1.11 12 Talc 1.11 General content 100.00 № p / p | Intragame 1 2 3 4 5 Solution with 6 7 8 Zero Ingredients Oleoylethanolamide Pantethine (dissolved) L-valine Microcrystalline cellulose pH 101 colloidal silicon dioxide binder Hypromellose 5 cps Isopropyl alcohol dichloromethane % by weight 42.55 31.91 2, 13 9, 04 8.51 0.53 in sufficient quantity in sufficient quantity Extragranular Ingredients 9 colloidal silicon dioxide 2, 66 10 magnesium stearate 2, 66 General content 100.00

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 61.54 2 Pantethine (dissolved) 23, 08 3 L-valine 7, 69 4 Microcrystalline cellulose pH 101 1.85 5 colloidal silicon dioxide 3, 08

- 17 040171

No. p / p Ingredients % by weight Solution with binder 10 Hypromellose 5 cps 0.62 eleven Isopropyl alcohol in sufficient quantity 12 dichloromethane in sufficient quantity Extragranular Ingredients 14 colloidal silicon dioxide 1.08 15 magnesium stearate 1, 08 General content 100.00

Example 17.

No. p / p Ingredients % by weight Intragranular Ingredients 1 Oleoylethanolamide 14.29 2 Pantethine (dissolved) 21.43 3 L-valine 57.14 4 Microcrystalline cellulose pH 101 1.57 5 colloidal silicon dioxide 3.57 binder solution 8 Hypromellose 5 cps 0.57 9 Isopropyl alcohol in sufficient quantity 10 dichloromethane in sufficient quantity Extragranular Ingredients 14 colloidal silicon dioxide 0.71 15 magnesium stearate 0.71 General content 100.00

Example 18 Stability and Dissolution Studies of the Formulations of Example 1

Stability condition. Accelerated Stability Study 40°C, 75% RH.

- 18 040171

No. researched Specification Study duration P/ Π nie Granulated Start 1 month 3 months 6 months powder from 1 Description white to almost white, filled into capsules sized Corresponds tvuet Reply there is Reply there is Reply there is "0" 2 Average capsule weight 550±7.5% 551.5 mg 550.9 mg 552.5 mg 550.7 mg 3 General content 450±7.5% 450.5 mg 452.8 mg 454.8 mg 452.7 mg 4 Time to break up the awns No more than 30 minutes 7-8 minutes 6-8 minutes 7-8 minutes 6-7 minutes 5 Analysis From 90.0% to — 5, 1 Analysis OEA 110.0% of declared content From 90.0% to 99.5% 100.7% 99.6% 98.9% 5, 2 Analysis of Lvaline 110.0% declared 98.6% 99.5% 100.2% 97.6% content 6 Dissolution 6, 1 Dissolved ie OEA Not less than 75% of the declared content 89.5% 87.7% 88.6% 88.9% 6, 2 Dissolved No less than ie L- valine 75% claimed content 83.6% 82.5% 89.2% 84.6%

- 19 040171 sti.

Example 19 Stability and Dissolution Studies of the Formulations of Example 12

Stability condition. Accelerated Stability Study 40°C, 75% RH

No. p/n Research e Specification Study duration 13 6 Start of months of months of months Granulated white powder 1 Description to almost white filled into capsules sized Corresponds tvuet Reply there is Reply there is Reply there is "0" 2 Average capsule weight 550±7.5% 553.7 mg 552.7 mg 553.8 mg 554.8 mg 3 General content 450±7.5% 451.8 mg 451.7 mg 453.5 mg 451.9 mg 4 Decay time No more than 30 minutes 7th minutes b-e minutes 7-8 minutes 6-7 minutes 5 Analysis From 90.0% to — 5.1 OEA analysis 110.0% declared 98.1% 99.6% 97.5% 97.1% content 5.2 Pantethine Analysis (50%) From 90.0% to 110.0% of declared content From 90.0% to 97.3% 97.6% 97.3% 98.1% 5.3 Analysis L- valine 110.0% declared content 97.4% 99.3% 100.0% 98.7% Analysis From 90.0% to 5.4 extract leaves 110.0% declared 91.4% 92.3% 93.1% 92.9% mulberries content 6 Dissolution Not less than 75% — 6, 1 OEA analysis declared 88.7% 86.8% 86.5% 87.1% content Dissolution Not less than 75% 6.2 pantethine declared 81.9% 80.1% 84.5% 85.7% (50%) content 6, 3 Dissolution L-valine Not less than 75% declared 84.8% 82.5% 84.3% 85.9% content 6, 4 Dissolution of mulberry leaf extract Not less than 70% within 60 minutes 89.2% 85.1% 83.3% 81.5%

- 20 040171

Example 20 Stability and Dissolution Study of the Formulations of Example 15

Stability condition. Accelerated Stability Study 40°C, 75% RH.

No. p/n Study Specification Study duration Start 1 months 3 months 6 months 1 Description White to off-white granular powder filled into "0" size capsules Corresponds Reply there is Reply there is Reply there is 2 Average capsule weight 570±7.5% 572.4 mg 571.8 mg 573.4 mg 571.6 mg 3 General content 470±7.5% 471.4 mg 473.7 mg 475.9 mg 471.4 mg 4 Disintegration time No more than 30 minutes 6-7 minutes 6-8 minutes 7-8 minutes 6-7 minutes 5 Analysis 5.1 OEA analysis From 90.0% to 110.0% declared content 98.2% 99.1% 98.4% 98.7% 5.2 Analysis of pantethine (dissolved o) From 90.0% to 110.0% declared content 97.1% 98.0% 98.1% 97.0% 5.3 Analysis of Lvaline From 90.0% to 110.0% of the declared content 99.5% 98.6% 99.3% 98.5% 6 Dissolution 6, 1 Dissolution OEA Not less than 75% of the declared content 88.4% 88.6% 89.5% 89.8% 6.2 Dissolution of pantethine (dissolved o) Not less than 75% of the declared content 81.1% 80.3% 82.1% 85.4% 6, 3 Dissolution L-valine Not less than 75% of the declared content 82.5% 83.4% 88.1% 85.5%

Example 21 Animal study.

Observation of the anti-obesity effect of a synergistic combination or composition/formulation containing OEA and other natural ingredients using a high-fat diet-induced obesity rat model. The following studies have been carried out:

male Wistar rats (Rat Rattus) (6 per group) were kept in a vivarium in a light/dark atmosphere on a 12-hour cycle at a temperature and relative humidity in the range of 1829°C and 30-70%, respectively. To maintain appropriate conditions, temperature and relative humidity were recorded three times daily. Throughout the experiment, the animals were fed standard rat food pellets (manufactured by VRK Nutritional Solutions, Pune) and ad libitum reverse osmosis water.

The composition at a normal diet with briquettes contained protein (18.1% v/v), fat (3.1% v/v), fiber (5.5% v/v), calcium (1.2% v/v) , phosphorus (0.5% v/v), total ash (5.5% v/v), carbohydrate (63% v/v) and moisture (7% v/v).

High fat diet composition contained normal diet with briquettes (36.5% v/v), soy powder (25% v/v), cheese (31% v/v), Dalda ghee (1% v/v) , Vitamin & Mineral Blend (6% v/v), DL-Methionine (0.3% v/v), Baking Powder (0.1% v/v), Sodium Chloride (0.1% v/v), supplied from the domestic market.

To evaluate the anti-obesity effect, forty-eight (48) male rats were screened and divided into eight groups. For comparative analysis, the groups were subdivided as follows:

normal control, disease control, individual component treatment groups, reference drug, and test composition/formulation of the present invention. In table. 1 provides details of the various groups and treatments used in the study.

Table 1

No. p / p Group Number of animals Test drug (mg/kg, orally) 1. G1 (normal control) 6 Normal control 2. G2 (disease control) 6 Disease control 3. G3 (OEA) 6 OEA (41 mg/kg) 4. G4 (L-valine) 6 L-valine (2 mg/kg) 5. G5 (pantethine) 6 Pantethine (31 mg/kg) 6. G6 (OEA+L-valine) 6 OEA (41 mg/kg) + L-valine (2 mg/kg) (Example 1) 7. G7 [Test composition (OEA+Lvaline+pantethine)] 6 OEA (41 mg/kg) + L-valine (2 mg/kg) + pantethine (31 mg/kg) (Example 15) 8. G8 (reference drug) 6 Reference drug (Orlistat 10 mg/kg)

treatment protocol.

The test animals were examined over a study period of 12 weeks. Animals in group (G1) were fed a normal briquette diet while the other groups were fed a high fat diet. After completion of the 12-week study, each group was analyzed for body weight and other evaluation parameters such as serum biochemistry (triglycerides, total cholesterol, and low-density lipoprotein (LDL) cholesterol), analysis of liver homogenates, and liver histopathology.

In table. 2 shows an analysis of the body weight measured in different groups.

In table. 3 is a summary of the results obtained from serum biochemistry in terms of % reduction in triglycerides, total cholesterol, and LDL.

In table. Table 4 describes a summary of the results obtained from liver biochemistry in terms of % reduction in triglycerides and total cholesterol.

table 2

Group No. Group ID Body weight G % body weight gain G1 Normal control 276.00 0.00 G2 Disease control 340.17 23.25 G3 OEA 310.00 12.32 G4 L-valine 338.67 22.71 G5 Pantethine 327.83 18.78 G6 OEA+L-valine 316.67 14.73 G7 OEA+L-valine+pantethine 293.33 6.28 G8 Orlistat 298.00 7.97

All the above values are averages.

Table 3

Group No. Group ID Triglycerides total cholesterol LDL (mg/dl) % lower NIA (mg/dl) % down NIA (mg/dl) % lower NIA G1 Normal 70.24 - 79.71 - 21.23 - G2 Disease control 122.78 - 147.24 - 57.97 - G3 OEA 100.05 18.51 105.54 28.32 35.54 38.70 G4 L-valine 115.23 6, 15 140.00 4.92 55.45 4.35 G5 Pantethine 110.13 10.30 135.54 7.95 49.17 15, 19 G6 OEA+L-valine 96.22 21.63 103.22 29.90 33.32 42.53 G7 OEA+L- valine + pantethine 73.98 39.75 80.55 45.29 22.52 61, 16 G8 Orlistat 77.62 36.78 84.27 42.77 25.74 55, 60

All values above are averages.

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Table 4

Group No. Group ID Triglycerides total cholesterol (mg/dl) % reduction (mg/dl) % reduction G1 Normal control 40.23 - 56.21 - G2 Disease control 83.00 - 90.25 - G3 OEA 59.98 27.74 74.12 17.87 G4 L-valine 80.02 3.59 85.23 5.56 G5 Pantethine 75.22 9.38 82.41 8.68 G6 OEA+L-valine 55.33 33.34 68.32 24.30 G7 OEA+L-valine + Pantethine 43.84 47.18 57.22 36, 60 G8 Orlistat 44.82 46.00 60.33 33.15

All values above are averages.

Interpretation and conclusion.

Obesity is essentially caused by an excess accumulation of fatty tissue, which leads to many disorders such as, but not limited to, insulin resistance, diabetes, fatty liver disease, coronary insufficiency, hypertension, and polycystic ovary syndrome. In obese humans, levels of total cholesterol, triglycerides, and LDL cholesterol fall outside the normal ranges. Similarly, the high fat diet rat model is a universally accepted model for investigating the efficacy of a drug in the treatment of obesity. In the experimental procedure presented above, obesity was induced by introducing a high fat diet (HFFA) over a period of 12 weeks. In comparison, animals in the normal control group were given a normal briquette diet, while the rest of the groups were given RVF.

As a routine experiment, the body weight of all animals was monitored weekly. In addition, various serum biochemistry parameters such as triglycerides, total cholesterol and LDL cholesterol were analyzed at the end of the 12 week treatment period. In this respect, at the end of the study protocol, all animals were sacrificed and the liver isolated. The liver homogenate of each individual animal was prepared and the level of total cholesterol and triglycerides was determined on it. In addition, a portion of the liver was used for histopathological studies.

Body weight analysis.

The main observation during the experiment was the monitoring of the body weight of the animals. Although the animals were examined weekly for change in body weight, mean body weight was measured at the end of 12 weeks and tabulated. 2. It is clear that the mean body weight of the disease control group (G2) was significantly higher (23.25%) compared to the normal group (G1). The test composition (G7) showed a tendency to reduce weight gain in an effective manner due to the composition/formulation of the present invention. From the results presented in Table 2, which presents a well-designed study comparing the % body weight gain in various groups of animals and the effect of the administration of the test composition/formulation, it seems obvious that the test composition effectively reduces weight gain compared to other groups, including reference preparation (Fig. 1).

Pathophysiology research.

At the end of the 12-week period, there was a significant increase (p<0.001) in serum total cholesterol, triglycerides, and LDL-cholesterol in the disease control group compared to the normal control group, suggesting that obesity was successfully induced in the models during the study .

From the results presented in table. 3, there is a significant reduction (p<0.001) in serum total cholesterol, triglycerides and LDL cholesterol when the test composition (G7) was given to the treatment group compared to the disease control group (FIGS. 2-4). These results are indicative of the fact that the use of the combination of the test composition/composition (G7) for the treatment of obesity and related disorders in rats has a significant efficacy in contrast to the individual components indicated in groups G3-G6. The test formulation also showed improved results compared to the reference formulation.

In addition, after 12 weeks of treatment, the level of total cholesterol and triglycerides was also determined in the liver homogenate. As indicated in Table. 4, there was a reduction in triglycerides and total cholesterol observed in the liver homogenate for all treatment groups (G3-G8) compared to the disease control group (FIGS. 5-6). More specifically, it should be noted that there was a significant reduction (p<0.001) in total cholesterol and triglycerides in the liver when the treatment group was given the test composition (G7) compared to the disease control group. This suggests that treatment with the combination of OEA was more effective in lowering total cholesterol and triglycerides in the liver compared to single components such as

- 23 040171 is indicated in groups G3-G6. The test formulation also showed improved results compared to the reference formulation.

As is apparent from the summary of data in Table 3, the % reduction in serum triglycerides, total cholesterol, and LDL levels was comparatively different in terms of parameters measured. It was observed that the % reduction in triglycerides due to the effect of the drug administered separately in groups G3, G4, G5 and G6 was 18.51, 6.15, 10.30 and 21.63, respectively, while the test composition (G7) by of the present invention showed improved results with a 39.75% reduction in triglycerides. In addition, it was observed that the % reduction in total cholesterol due to the effect of the drug administered separately in groups G3, G4, G5 and G6 was 28.32, 4.92, 7.95 and 29.90, respectively, while the test composition (G7) of the present invention showed improved results with a 45.29% reduction in total cholesterol. Similarly, the % LDL reduction due to the effect of the drug administered alone in groups G3, G4, G5 and G6 was observed to be 38.70, 4.35, 15.19 and 42.53, respectively, while the test composition ( G7) of the present invention showed improved results with a 61.16% reduction in LDL levels.

Similarly, it was observed that in the case of liver histopathology, the level of steatosis was significantly lower in the case of the treatment group given the test composition (G7).

As is apparent from the summary of data in Table 4, the % reduction in hepatic triglycerides and total cholesterol had comparative differences in terms of measured parameters. It was observed that the % reduction in triglycerides due to the effect of the drug administered separately in groups G3, G4, G5 and G6 was 27.74, 3.59, 9.38 and 33.34, respectively, while the test composition (G7) by of the present invention showed improved results with a 47.18% reduction in triglycerides. In addition, it was observed that the % reduction in total cholesterol due to the effect of the drug administered separately in groups G3, G4, G5 and G6 was 17.87, 5.56, 8.68 and 24.30, respectively, while the test composition (G7) of the present invention showed improved results with a 36.60% reduction in total cholesterol.

Among the treatment groups, it was observed that the test composition (G7) exhibited the highest anti-obesity activity in rats given RVCF. A summary of the data indicating this is given in Table. 2-4 and in Fig. 1-6. From the study, it is clear that the test composition/formulation (G7) provides a synergistic effect compared to the individual components of the composition and the reference drug, and also plays an important role in the anti-obesity activity in rats that were given RVCF.

Moreover, no mortality was observed in any group of animals, suggesting that obesity did not lead to death, but increased the prevalence of the disease in animals.

Conclusion.

Based on an experimental animal study, it can be concluded that the test composition of the present invention was more effective and has a synergistic effect compared to a single drug for the treatment of rats with RVCA.

Based on the study, it was also observed that the pharmaceutical composition/formulation of the present invention was safe and effective in reducing body weight, triglyceride and cholesterol levels, and thus reduced the risk of obesity and associated fat metabolism disorders.

While the invention has been described and illustrated with reference to certain embodiments, one skilled in the art will appreciate that various adaptations, changes, modifications, substitutions, deletions, or additions of procedures and protocols can be made without departing from the spirit and scope of the invention. For example, effective dosages other than the specific dosages set forth herein above may be employed as a result of variation in the response of the mammal being treated to any indication of the compounds of the invention as set forth above. The observed specific pharmacological responses may vary according to and depending on the specific active compounds chosen or whether pharmaceutical carriers are present, as well as the type of formulation used and the mode of administration, such variations or differences being expected to be consistent with the objectives and practical implementation. of the present invention. Thus, it is intended that the invention be defined by the scope of the claims, and that such claims should be interpreted broadly as appropriate.

Claims (11)

CLAIM 1 . Pharmaceutical composition for the treatment of obesity and related disorders, containing a combination of: 42.55% by weight oleoylethanolamide (OEA), 2.13% by weight of valine, and - 24 040171 31.91% by weight pantethine. 2. The pharmaceutical composition of claim 1, wherein the composition further comprises a naturally occurring alpha-glucosidase inhibitor, a naturally occurring fatty acid amide hydrolase (FAAH) inhibitor, or a combination thereof. 3. The pharmaceutical composition according to claim 2, wherein the natural alpha-glucosidase inhibitor is 1-deoxynojirimycin. 4. The pharmaceutical composition according to claim 2, wherein the amount of the natural alpha-glucosidase inhibitor is in the range of 0.5 to 1.5% by weight of the composition. 5. The pharmaceutical composition of claim 2, wherein the natural FAAH inhibitor is selected from quercetin, myricetin, isorhamnetin, kaempferol, pritimerin, curcumin, biochanin A, genistein, and daidzein. 6. The pharmaceutical composition of claim 2, wherein the amount of the natural FAAH inhibitor is in the range of 0.5 to 25% by weight of the composition. 7. Pharmaceutical composition according to any one of the preceding claims, additionally containing pharmaceutically acceptable excipients. 8. The pharmaceutical composition according to claim 7, wherein the pharmaceutically acceptable excipients are selected from diluent, binder, lubricant, glidant and solvent. 9. The pharmaceutical composition according to claim 8, in which the amount of diluent is in the range from 1 to 30% by weight of the composition, the amount of binder is in the range from 0.5 to 1.5% by weight of the composition, the amount of lubricant is in the range of 0.5 to 5% by weight of the composition, the amount of glidant is in the range from 1.0 to 25% by weight of the composition, or the solvent is taken in sufficient quantity. 10. A pharmaceutical composition according to any one of the preceding claims, wherein the composition is in the form of a tablet, capsule, sachet, pill, liquid or solid filled hard capsule, soft capsule, powder, granule, suspension, solution or modified release formulation. 11. A method for producing a pharmaceutical composition according to any one of the preceding paragraphs, including: a. separate weighing of OEA, valine and pantethine in separate containers, b. sifting pre-weighed OEA, valine and pantethine separately, c. mixing content in step b, d. obtaining a binder solution in a separate container and adding it to step c to obtain a paste, and sieving the paste to obtain granules, e. drying the resulting granules to reduce the dryness level (DR) to less than 1.5% v/v, f. sieving the dried granules through a suitable sieve and g. screening the pre-weighed lubricant(s) or glidant(s) separately through a suitable sieve and adding to step f.