RS64163B1 - Sinergistic combination of glucomannan and monacolin and their use as non-toxic hypolipemics - Google Patents

Description

TECHNICAL FIELD

This invention relates to a natural hypolipemic preparation consisting of a combination of active substances isolated from natural material for use in the prophylaxis or treatment of increased blood fats, cholesterol, triglycerides and related to atherosclerosis, elevated blood pressure and other diseases caused by metabolic disorders, inhibiting HMG-CoA reductase, as well as a procedure for their preparation. According to the IPC, the invention belongs to classes A23L31/00, A61K31/736, A61K36/888, A61K36/899, A61P1/00.

TECHNICAL PROBLEM

This invention solves the problem of obtaining a natural, non-toxic and safe preparation, which reduces the level of fat in the blood and which has a pronounced activity against metabolic disorders and diseases caused by such disorders. The subject invention unexpectedly corrects metabolic disorders, reduces the level of fat in the blood, without producing the known toxic manifestations of previously known preparations containing the same active principle.

The invention provides an effective, safe, non-toxic and economical natural product which as such can be used as a dietary supplement for preventive purposes, but also for the suppression of diseases caused by metabolic disorders and the increased level of fat caused by them, both in adults and in children.

It is believed that the alarming increase in the incidence of cardiovascular diseases is the result of the modern way of eating, which is characterized by an increase in the consumption of refined fats and carbohydrates, with a simultaneous decrease in the consumption of polyunsaturated fatty acids and fibers. The most common problems caused by such a diet, which leads to metabolic syndrome, are elevated blood cholesterol levels, hypertension, dyslipidemia, elevated blood pressure, or diseases such as diabetes, cardiovascular problems, and hepatic steatosis.

Numerous epidemiological and experimental studies have shown that metabolic syndrome affects almost half of the population of adults over 50 years of age. It is believed that this incidence will continue to increase in the future, given the prevalence of obesity in early childhood. Excess body fat, especially when it is localized in the abdominal area, leads to an imbalance of fat and sugar metabolism that induces hyperinsulinemia (a high level of insulin in the blood as an indicator of increased resistance to this hormone due to increased pancreatic secretion caused by cell resistance to sugar intake).

Modern drugs used in the treatment of hypercholesterolemia are statins (HMG-CoA reductase inhibitors) and fibrates (most useful in the presence of high triglycerides). However, the use of these drugs, it is now completely certain, causes a large number of side effects. Statins have numerous serious side effects, the most serious of which are myopathy, hepatotoxicity and liver damage as well as dermatomyositis, and on laboratory blood tests, this is visible in the form of elevated levels of transaminases, alkaline phosphatase, γ-glutamyl transpeptidase, bilirubin and abnormalities of the thyroid gland function.

Due to the serious toxicity of antilipemic drugs, there is a need to find a safer therapy for lowering blood fat levels. In addition to strategies for combating metabolic disorders, based on lifestyle and diet changes, the use of natural substances with beneficial properties is also common. An alternative strategy of classic pharmacological intervention is the use of nutraceuticals with a good tolerance profile and activity on the lipid profile.

A good effect on cholesterol synthesis is e.g. policosanol. It decreases HMG-CoA-reductase expression in a concentration-independent manner, possibly through receptor-mediated mechanisms that inhibit transcription for this enzyme. Some studies have shown that policosanol induces other effects on cardiovascular risk, similar to the pleiotropic effects attributed to statins.

It is also known that one type of monacolin, which is part of red fermented rice, monacolin-K, is an inhibitor of HMG-CoA reductase, an enzyme that participates in the synthesis of cholesterol in the liver, and is known in the pharmaceutical industry as mevinolin or lovastatin. It is capable of inhibiting HMG-CoA reductase, which is a key enzyme in cholesterol biosynthesis. Cholesterol synthesis is inhibited by monacolins, which are metabolites extracted from fermented rice with the help of a special type of Monascus purpureus yeast. 15 types of monacolin are known and all of them have lower or greater antilipemic activity, the most active of which is monacolin K (mevinolin or lovastatin).

Red rice contains up to 5% monacolin K, which is a natural substance that has been used in China for thousands of years to reduce fat. However, monacolins, which are structurally statins, although to a lesser extent than the purified individual synthetic pharmaceutical statin preparations, themselves cause unwanted effects. However, their use as nutritional supplements, unlike prescription drugs, carries an increased risk due to the accumulation of side effects, which makes them a group of nutraceuticals that are not safe and require serious monitoring of the effects of their action.

It is also known that glucomannan (3 g per day) is particularly useful in reducing body weight and lipid levels, but also for constipation, i.e. regulation of bowel function as well as in irritable bowel syndrome. Glucomannan is a high molecular weight polysaccharide extracted from the root of the Amorphophallus konjac plant, which is used in Japanese cuisine as a gelling agent. This fiber has the ability to bind to itself a large amount of water, increasing the volume from 60 to 200 times and giving a soft gelatinous mass. This feature gives glucomannan a double beneficial effect, binding and expelling excess water from the body and causing a feeling of satiety.

There are preparations on the market for lowering blood lipid levels, which, in addition to fermented red rice extract, also contain other active substances such as plant fibers, artichoke, policosanol and the like, which through their synergistic action enhance the effect of monacolin K. However, all these preparations are products that require monitoring of monacolin side effects due to their toxicity.

The need for new natural substances and compositions with which it will be possible to effectively prevent and treat metabolic disorders that result in an increase in the level of fat in the blood, and other diseases that are related to it, without having the side effects that the known preparations have, is a serious technical problem faced by modern pharmaceutical science, and has led to the subject of this invention. A special challenge is obtaining an economical natural product with a proven and safe pharmacological effect.

The subject invention is particularly useful in patients with poor tolerance to traditional medicines or those who reject traditional medicines, and it achieves a solution to the problem of obtaining a natural, economical, effective and harmless preparation for the treatment of metabolic disorders and lowering the increased level of lipids in the blood, as well as diseases caused by these disorders.

STATE OF THE ART

The use of red fermented rice was first documented as early as 800 AD in China during the Tang Dynasty, and is described in detail in the Chinese Pharmacopoeia. According to those records, red fermented rice was recommended for digestive and intestinal problems, and for blood circulation, and red rice powder, known as Zhi Tai, but also as an alcoholic extract - Xue Zhi Kang, was used. Red rice is still used today in traditional Chinese cuisine as an addition to dishes, drinks, cosmetics, but also as a natural preservative, color, spice and raw material for rice wine.

In the late 1970s, researchers in the United States and Japan isolated lovastatin first from Aspergillus and monacolin from Monascus. Chemical analysis soon showed that lovastatin and one of the monacolins - monacolin K, are identical in terms of their chemical structure.

Monacolin K, a natural statin - lovastatin, is produced by mycotic fermentation of rice by Monascus purpureus and is an inhibitor of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase. In China, it has been a traditional medicine for reducing fat for centuries.

Red fermented rice obtained by fermentation with the fungus Monascus purpureus as a dietary supplement is recommended for people with elevated cholesterol. Its effectiveness, either as an independent food supplement or in combination with other preparations, 20 has been examined in more than 30 clinical studies available in the PubMed database. In almost all studies, a significant decrease in the concentration of total cholesterol, as well as "bad" LDL-cholesterol, was observed in those patients who took the preparation compared to placebo.

One of those works published on PUB MED; Duodecim.; 2010;126(6):623-6. Enkovaara AL reports that preparations based on red rice containing about 5 to 10 mg of lovastatin at the daily dose recommended on the package can reduce blood cholesterol levels but show similar drawbacks and interactions as statin drugs.

Preparations containing red rice contain various active substances, but the most important for their action are monacolins, which block the synthesis of cholesterol in the liver. One of the monacolins - monacolin K, whose chemical structure is identical to lovastatin, a drug from the statin group that is prescribed for the treatment of high cholesterol, has the same structure and the same effect in the body.

Different species of Monascus fungi will produce different amounts and types of monacolin. The "Vent" strain of Monascus purpureus, when properly fermented and processed, yields a red rice powder containing about 0.4% monacolin, of which approximately half will be monacolin K (identical to lovastatin).

The safety of fermented rice products has not been determined, among other things, because commercial supplements have highly variable amounts of monacolin and often contain high levels of the toxin citrinin without being precisely declared, so defining risks is difficult and this makes these products risky. On the other hand, large amounts of pure monacolin K in red rice extract indicate the enrichment of such an otherwise natural product with synthetic, purified lovastatin.

Due to all of the above, nutritional supplements - preparations containing red rice extract - carry the risk of side effects and interactions characteristic of drugs from the statin group, especially in patients who have previously experienced side effects of statins such as muscle and liver damage.

Today, it is known that red fermented rice, which, among other things, contains monacolin K, inhibits HMG-CoA reductase, reduces the level of total cholesterol, triglycerides and reduces the level of LDL and increases the level of HDL cholesterol in the blood, helps digestion and as such, is used as a safe food additive, is used to color and improve various food products, is taken to strengthen the body, helps digestion and blood circulation, spleen health and stops diarrhea.

One of the published studies showed in controlled trials that red rice works very well, leading to a 22% reduction in total cholesterol, LDL cholesterol by 28%, triglycerides by 32%, while HDL cholesterol concentration increased by 20% after only 8 weeks of use.

A 2005 study published in the "European Journal of Endocrinology" showed that taking 600 mg of red fermented rice twice a day caused an average reduction in LDL cholesterol of 26% and total cholesterol was reduced by an average of 20% after 8 weeks. Lovastatin is more potent than red fermented rice because according to the prescribing information for Mevacor®, lovastatin reduces total cholesterol by 16 to 24% and LDL cholesterol by 21 to 32%, depending on the daily dose. In a meta-analysis of 91 randomized clinical trials with a duration of 12 weeks, a total of 68,485 participants, LDL-cholesterol was lowered by 24 - 49% depending on the statin used.

Despite all of the above, red fermented rice has become a popular over-the-counter supplement, i.e. food supplement, due to the fact that its effect cannot be much different from the effect of lovastatin or other statin drugs, which have proven harmful effects. In the US, therefore, red rice extracts containing lovastatin were classified as drugs by the US FDA and removed from the market.

Lovastatin is a cholesterol-lowering drug from the statin group of drugs. It was the second drug of this class to be discovered. The drug was discovered by Merck in 1978 after testing 18 compounds. Lovastatin is twice as potent as its predecessor, mevastatin. It is structurally similar to hydroxymethylglutarate (HMG), a substitute for HMG-coenzyme A (HMG-CoA), a substrate in the cholesterol biosynthetic pathway. Lovastatin is a competitive inhibitor of HMG-CoA reductase with 20,000 times greater binding affinity than HMG-CoA. Lovastatin differs structurally from meVastatin by one methyl group in the 6' position. It is a prodrug that is activated in vivo by hydrolysis of the lactone ring.

Lovastatin is a prescription drug, the dosage is standardized and regulated by the FDA, so each batch has the same amount of drug and the dosage for lovastatin is 20 mg per day with a maximum daily dose of 80 mg.

Red fermented rice is a dietary supplement, and is not standardized as a drug. According to research conducted by Natural Standard, the amount of monacolin in the tested capsules varies from 0.15 to 3.37 mg per capsule.

Although it is generally well tolerated by most people, one of the more serious side effects of red yeast extract is the development of muscle pain as a result of the destruction of muscle tissue - rhabdomyolysis. Symptoms include muscle pain, tenderness and weakness. The destruction of muscle tissue can lead to kidney damage over time and to their failure (renal failure). The risk is higher in those who take high doses of this extract or take it together with lovastatin.

Red fermented rice extract may also cause liver damage. Symptoms of liver damage include yellowing of the eyes and skin, dark urine and elevated liver enzymes. Allergic reactions have also been reported with the use of red yeast extract, including itching, difficulty breathing, swelling of the face, tongue, and throat, along with difficulty swallowing, gastrointestinal side effects, gas, and bloating.

Medline Plus and Memorial Sloan Kettering Cancer Center warn that red yeast that is not properly fermented contains a substance, citrinin, which is toxic to the kidneys and liver.

However, in 2011, EFSA granted approval for monacolin K from fermented red rice (Joumal 2011 ;9(7):2304) to normalize LDL cholesterol.

On the other hand, glucomannan - propol mannan is a mega-molecule extract from the Amorphophallus konjac plant Amorphophallus propol from the Konjac plant family and it is the most concentrated known fiber that enables weight loss while maintaining satiety. It has been used in Japan for over 300 years to achieve balance in the human body. It is a natural substance with the highest molecular weight and its effect has been clinically proven at the pharmaceutical level, that it works to reduce body weight in a healthy and natural way, by eliminating fat through physical means, and leads to a reduction of up to 2.5 kg of pure fat per month, a reduction in the level of LDL ("bad" cholesterol) in the blood, a reduction in the level of triglycerides in the blood, but also a reduction and stabilization of the blood sugar level. It also has a probiotic effect, improving the health of the intestinal flora and has a strong anti-constipation effect thanks to better intestinal peristalsis.

Propol mannan binds dietary fat in the body, controls the glycemic index (GI) and keeps it at a low level, increases intestinal efficiency and contributes to better utilization of food, gives a feeling of prolonged satiety, lowers bad cholesterol, and regulates blood sugar levels.

As such, it received approval from the European Food Safety Authority in 2010 (EFSA Journal 2010;8(10):1798), and the health claims related to konjac mannan - glucomannan, i.e. propol mannan, are weight reduction, reduction of glycemic response after a meal, maintenance of normal blood glucose concentration, maintenance of normal blood triglyceride concentration, maintenance of normal cholesterol concentration, maintenance of normal intestinal function and reduction of potentially pathogenic microorganisms in the intestines.

EFSA Joumal 2009;7(9).T258 approval for glucomannan relates to the normalization of blood cholesterol and its effect on improving human health.

Konjac glucomannan are fibers of high molecular weight soluble in water, which after hydration form a highly viscous gel that increases the initial volume by up to 200 times, acts as a magnet for binding fats and prevents dietary fats from being absorbed in the gastrointestinal tract, and also shows a glandular effect in the digestive tract because it increases the excretion of bile and excess bile is removed through feces and blocks the reabsorption of cholesterol. Konjac mannan, Konjac glucomannan is a slightly branched polysaccharide consisting of (3 -1,4 linked D-glucose and D-mannose in a ratio of approximately 1 to 1.6. The average molecular weight is 200 - 2000 kDa and this makes it a macromolecule. It is used in food, disperses<U>in cold or warm water and is a source of dietary fiber. It is usually made from the roots of the Konjac plant. 30 Glucommannan powder, capsules and tablets are used as a medicine. Glucomannan is used for weight loss, high cholesterol, high blood pressure and stomach disorder called dumping syndrome. In food, glucomannan is used as a thickening agent or gelling agent. Glucomannan flour and powder are used for food production.

The beneficial properties of the Konjac plant have been known since ancient times. Its dietary fibers can collect up to 10 times more fat and water than fibers normally used in food and expel them from the body through the stool. That is why glucomannan is effective in lowering body weight as well as blood sugar and cholesterol.

In the world, glucomannan is used in the form of nutritional supplements (tablets and capsules), then soup in a bag, meal replacement shakes such as powdered food, coffee to reduce the feeling of hunger, various drinks, biscuits and crackers to achieve suspension of hunger. Body weight and body health are regulated by controlling and preventing overeating. Over 100 clinical studies have documented the aforementioned activities of propol mannan.

Both active substances have EFSA approval.

There are papers that talk about the effectiveness of monacolin K from red fermented rice and glucomannan, associated with a low-calorie diet, in improving the lipid profile and hypertension, a symptom of metabolic syndrome.

In the patent state of the art there are the following patents and applications related to glucomannans and monacolins:

The most relevant literature related to this topic is the capsule formulation described by patent CN106822503 that contains purified powder of the konjac plant and powder and extract of red rice to reduce serum lipids that inhibits the formation of atherosclerotic plaque, protects vascular endothelial cells and inhibits lipid deposition in the liver. The application protects the formulation of the capsule, but this formulation did not solve the problem of toxicity.

CN106889605 is a patent that describes a composition composed of red fermented rice extract and konjac glucomannan, as well as numerous other herbal active ingredients from traditional Chinese medicine that are used as a medicine to reduce blood fat, improve HDL-c values with the effect of lowering blood pressure and normalizing glucose metabolism.

Application W02018/007820 describes a probiotic with one or more Lactobacillus strains WITH the addition of statins, stanols and sterols, monacolin K, fatty acids, niacin, soluble fiber and/or prebiotics, from the statins fluvastatin, pravastatin, simvastatin, atorvastatin and/or rosuvastatin but without lovastatin.

The invention described in the patent CN103110103 provides a nutritional replacement for any meal in the form of a powder that is prepared by adding hot water, which contains, in addition to konjac glucomannan and red fermented rice extract, collagen, barley, lentinan powder, oats, lotus, calcium lactate, salt and is successfully applied to remove excess fat and lose weight, treat fatty liver, reduce blood pressure, blood sugar levels, increase bone density, improve hair quality, immunity and similarly.

CN105146572 describes an invention that relates to the processing and condensation of meat in the production of sausages with the addition, among other ingredients, of red fermented rice extract and konjac glucomannan.

CN105285702 describes an invention for preparing wheat pastry with the addition of glucomannan and red fermented rice, which strengthens the hematopoietic function of the human body and improves blood circulation.

Patent JPH10114682 describes the formulation of a drug that, in addition to the active substance (chemotherapeutic drug, antibiotic, antipyretic analgesic, anti-inflammatory agent and antipsychotic), also contains glucomannan in pharmaceutical form and does not refer to a combination with monacolin K or any other antilipemic drug.

In the patent state of the art, there are patents related to formulations containing glucomannan and lovastatin, but they also contain other active substances such as niacin (Vitamin VZ), Bacillus coagulans, cyclodextrin, in various forms for the treatment of obesity, fibrosis, etc., but not a single patent has been found that refers to the combination of only active substances glucomannan and a mixture of monacolin with a certain amount of monacolin K (lovastatin), not in any pharmaceutical form, nor for use as an antilipemic, and especially not that the problem of monacolin toxicity was solved. A large number of patents have been found that refer to formulations that, among other active substances, may contain either glucomannan or monacolin, which are known both in the literature and in traditional Chinese medicine and cuisine, but not a single patent refers to the synergistic action of these active substances in the treatment of hyperlipemia, without adverse toxic effects.

The work published in 2015 in the journal Biology and Medicine under the title Effects of Monacolin K of Red Rice and Glucomannan, Combined with a Low Calorie Diet, in Treatment of Dyslipidemia and Hypertension Esposito et al., describes the subject and the results of the research from which it can be seen that the combination of active substances monacolin K from red fermented rice and glucomannan, which are known as extremely active antilipemics, has an effect on reducing lipids but with the application of a low-calorie diet.

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However, this work did not eliminate the fear of monacolin K side effects, nor does the work deal with this problem.

In the literature, the investigation of the enhanced effect of statin action with the addition of glucomannan has already been described. In the book Combination TĆegaru in Dyslipidemia by Maciej Banach, published by Springer Intemational Publishing: Imprint: Addis, 2015, on page 204, it is stated that clinical studies have been conducted that have shown that the use of soluble fiber, glucomannan, plant sterols and probiotics inhibits the absorption of cholesterol and bile salts, and there are clinical studies that show that these nutrients have additional effects on statins.

Known works refer to the use of two active substances, but their application is limited by toxicity and side effects caused by monacolins - statins.

In addition to all the mentioned literature and patent data, the use of red rice extract in combination with other active ingredients is evidently limited by the toxicity of such preparations according to the state of the art.

DESCRIPTION OF THE INVENTION

The invention described here refers to a natural food supplement based on a combination of highly purified raw materials konjac glucomannan and fermented red rice, with added sugar and/or sweeteners, which has a synergistic antilipemic effect, without harmful and hepatotoxic effects. These two active ingredients in combination strongly lower cholesterol levels, by strongly suppressing synthesis and rapidly increasing cholesterol excretion. The overall synergistic effect is the improvement of the lipid status of the user of this product, which can be used preventively but also as an addition to already existing hyperlipidemic therapy, all without the hitherto known toxic effect, in a total daily dose of up to 10 mg of monacolin K. The effects of applying the stated daily dose were examined on 27 subjects. The effects of the dose of the active substance monacolin K applied in this way, 5 mg twice a day, which are within the scope of the effects of the usual pharmacotherapy that is applied in multiple doses of the active substance, unequivocally prove the synergistic effect of the combination of the described natural preparation, which contains, in addition to monacolin K, glucomannan. The invention is not limited to the stated dosage, but it is also possible to use larger doses in accordance with the prescribed maximum daily doses in pharmaceuticals, which is up to 80 mg of lovastatin, i.e. monacolin K.

According to the present invention, the procedure for obtaining a safe product for reducing the level of lipids in the serum containing red fermented rice extract and glucomannan consists in the use of a highly purified dry extract of red fermented rice from Monascus purpureus where the monacolin content is defined as containing at least 0.5% of monacolin K but which does not contain monacolin A - citrinin in an amount greater than the detection limit of the method, which is 15 pg. And the second active component of this combination of active substances, plant fiber glucomannan, is highly purified propol mannan obtained from the plant Amorphophallus Propol (Konjak Koch) without the detected presence of sulfur dioxide and arsenic.

Some species of the fungus Monascus purpureus produce the toxic monacolin A known as citrinin, which can cause kidney and liver damage if found in such a natural mixture. Citrinin is a mycotoxin that is often found in food and causes various toxic effects, nephrotoxic, hepatotoxic, cytotoxic.

Citrinin was one of many mycotoxins discovered in 1930 by H. Raistrick and AC Hetherington. In 1941, H. Raistrick and G. Smith identified citrinin as a substance with broad antibacterial activity. However, in 1946, Ambrose and F. De Eds showed that citrinin was toxic to mammals. In 1948, the chemical structure was solved by V.B. Vhallei et al.

Citrinin is a natural compound, first isolated from the fungus Penicillium citrinum, after which it got its name, but it is also produced by other types of fungi Penicillium, Monascus and Aspergillus. In 1993, the International Agency for Research on Cancer of the World Health Organization began to evaluate the carcinogenic potential of mycotoxins and maximum levels of unwanted substances in animal feed were prescribed in the EU Directive of the European Parliament and of the Council of May 7, 2002, but no maximum level has yet been determined for citrinin in feed.

Citrinin is a polyketide mycotoxin, which is a secondary metabolite of some types of fungi. Its IUPAC name is (3R,4S)-4,6-dihydro-8-hydroxy-3,4,5-trimethyl-6-oxo-3H-2-benzopyran-7-carboxylic acid, the molecular formula is C13H14O5, and it is hardly soluble in cold water. The mechanism of action of citrinin in the body has not yet been clarified, and its intermediates during biotransformation are also unknown.

The types of toxicity that have been studied and confirmed for citrinin are acute toxicity, nephrotoxicity, genotoxicity and its carcinogenicity.

The acute toxicity of citrinin depends on the route of administration and the species used for research. Oral administration has the highest lethal dose, and the LD50 of this administration is e.g. for rabbits it is 134 mg/kg body weight while intravenous administration defines the lowest lethal dose of LD50 19 mg/kg. Across crops, the LD50 is 57 mg/kg body weight for ducklings.

The composition of the formulation that is the subject of this invention includes a highly purified red fermented rice extract that was tested by the VICAM Citri test LC-MS/MS method (LOQ 15 µg/kg) and showed a value of citrinin <15 µg/kg, i.e. citrinin was not detected.

Also, the level of allergen (gluten), determined by the R-blOpHARM ELISA method (LOQ 5 mg/kg) is below the detection limit, i.e. < 5 mg/kg.

The novelty of this invention refers to the evidence documented here that such highly purified monacolins and glucomannan have a pronounced synergistic antilipemic effect without increasing liver enzymes as an indicator of hepatotoxicity, which is usually present when these substances are used in known formulations.

There is a theory that additional substances naturally present in red rice extract, or mixtures of natural statins, enhance the action of monacolin and that this is the advantage of these preparations compared to prescription drugs (individual statins), but also that they contribute to reduced toxicity. There is currently no scientific evidence to support this theory.

The invention in question provides a new argument in support of such claims, because in the clinical trial that was conducted, by controlling the liver enzymes of volunteers, it was established that there was no increase in this toxicity indicator beyond the limit of normal values, which

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confirms that the product described in this patent has no unwanted toxic effects, although it consists of a mixture of natural statins - monacolin, which contains at least 0.5% of monacolin K, purified, in which monacolin A is not detected - citrinin and purified plant fibers glucomannan, in the form of propol mannan in which arsenic and sulfur dioxide are not detected.

According to the invention, the therapeutic bioactivity of the product formulated in this way can be used for the prophylaxis and treatment of metabolic disorders that result in an increased amount of lipids in the blood serum, because they have a strong inhibitory power on fat synthesis and act on increased fat excretion.

Testing on volunteers confirmed an increase in the activity of the preparation formulated in this way compared to previously known ones, without any toxic manifestations being recorded.

The invention is based on basic evidence that in subjects who did not take other antilipemic therapy and did not change their lifestyle or diet, using only the product that is the subject of this invention, which consists of two active substances, fermented red rice extract and konjac glucomannan, there was a decrease in the level of total cholesterol, a decrease in LDL cholesterol, an increase in HDL cholesterol, a decrease in triglycerides, a decrease in body weight, a decrease in body fat, a decrease in blood sugar concentration, without an increase, above permitted values, toxicity indicators that were examined, namely gamma glutamyl transferase and creatine kinase. The antilipemic effect was confirmed without toxic manifestations that are usually characteristic of the use of the mentioned active substances. According to the subject invention, it was established that the same effect of the tested preparation was achieved by using a dose that was several times smaller than the pharmaceutical lovastatin. This further confirms the synergistic effect of the combination of glucomannan and red fermented rice monacolin. This is significant because the same effect is achieved by using a much smaller amount of the active statin substance, which further reduces the harmful effects of using these compounds.

According to the invention, it was shown that the extract of red fermented rice in combination with glucomannan is a highly potent inhibitor of HMG-CoA reductase, without accompanying toxic effect. Among the side effects, sporadic spasms of low intensity, in rare cases difficult emptying or nausea, rarely flatulence or abdominal pain and the appearance of gas were recorded, but none of the listed sporadic side effects can be directly related to the use of the preparation, and accordingly, none of the subjects left the trial due to any of the side effects of the trial.

It was also observed that the effect of applying the product, which is the subject of the invention, is proportional to the initial values of cholesterol and triglycerides.

BEST MODE FOR APPLICATION OF THE INVENTION

Due to all of the above, the product containing red rice extract in combination with glucomannan, which are the subject of this invention, are effective in preventing and/or treating metabolic disorders and hyperlipemia, and therefore high blood pressure, atherosclerosis and other diseases caused by increased levels of lipids in the blood.

The invention relates to obtaining and using a natural product as a prophylactic and therapeutic agent (food additive) and can be applied to both adult patients, due to its proven safety, and children.

Interventional testing of the effectiveness of the product that is the subject of the invention was performed on 27 subjects (12 women and 15 men) with an average age of 55 years, with the youngest subject being 33 and the oldest 63 years old. In the group there were 1.4% undernourished, 5.19% normally nourished, 9.3% obese and 85% overweight subjects. 4 control examinations were performed for 15 days with the recommended intake of 2 sachets of the preparation in two equal daily doses of 5 g of the oral powder preparation 15-20 minutes before lunch and dinner with plenty of water or as a meal replacement. No one was taking cholesterol-lowering therapy, and the study was conducted with the subjects' usual diet, without dieting and increasing physical activity.

Anthropometric parameters were monitored on the InBody 720 device, waist and hip circumference, blood pressure, and biochemical blood analyzes were performed at the beginning and end of the study. The parameters examined are glucose, cholesterol, HDL, LDL, triglycerides, urea, creatinine, uric acid, total bilirubin, ALT, AST, gamma GT, creatinine kinase.

The following examples illustrate the inventive concept, in accordance with the description of the invention indicated in this patent, in terms of the formulation and effects of the application of the invention.

The foregoing examples provide illustration of the invention, and nothing herein should be construed as limiting the overall scope of the invention.

The invention will be further illustrated by non-limiting examples.

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EXAMPLES

Example 1 - Formulation of a powder composition for oral administration

40 kg of highly purified konjac glucomannan (without detectable sulfur dioxide and arsenic) is mixed with purified dry extract of fermented red rice (Monascus purpureus) in an amount that ensures a content per dose of monacolin K of at least 5 mg (without detectable citrinin) and 50 kg of a mixture of excipients containing 5-50%, glucose monohydrate as a sweetener, 5- 35%, acidity regulator, sodium hydrogencarbonate and citric acid, aroma, 1 - 5% magnesium oxide anti-caking agent and steviol glycoside as an additional sweetener. The powder formulated in this way is stored at a temperature of 15 - 25 °C and used for various oral dosage forms. 5 g of the powder formulated in this way contains a minimum amount of 5 mg of monacolin K and is suitable for formulating a unit dose that can also be a meal replacement.

Example 2 - Change in body weight (kg)

Table 1. - Change in body weight (kg)

The test results from Table 1 show that, with the use of the product that is the subject of this patent, the subjects lost an average body weight of 1.04 kg in 45 days of use.

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Example 3 - Change in body fat (kg)

Table 2. - Change in body fat (kg)

The test results listed in Table 2 show that, with the use of the product that is the subject of this patent, the subjects lost an average of 1.23 kg of body fat in 45 days of use.

Example 4 - Weight loss expressed as a percentage of subjects

Table 3. - average number of respondents who lost weight in different ranges

The listed results from Table 3 show that by applying the product that is the subject of this patent, 2/3 of the subjects lost body weight in 45 days of application, with 22% of the subjects losing twice as much as the average.

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Example 5 - Examination of blood parameters

Table 4. - Results of blood parameters tests at the beginning and at the end of the study on all 27 subjects

Gamma-GT - y-glutamyltransferase or gamma glutamyl transferase (GGT) or gamma glutamyl transpeptidase (GGTP) is a membrane enzyme from the group of peptidases that catalyzes the transfer of a glutamic acid residue released by hydrolysis to an amino acid or peptide and is also involved in glutathione metabolism. In the body, it occurs predominantly bound to cell membranes. The most gamma glutamyl transferase is found in the kidneys, prostate, liver, small intestine epithelium and brain, and is found in places of intensive absorption of amino acids.

Although GGT is also present in the pancreas and kidneys, an increase in serum GGT catalytic activity is actually the most sensitive indicator of liver damage. This enzyme is considered the most sensitive indicator of hepatobiliary disease, because even a small disruption of cellular integrity or minimal cholestasis leads to a significant increase in gamma GT activity in the serum. Normal values differ from the gender and age of the examinee, but from 8 to 65 units of GGT per liter of blood (u/L) can be considered normal values.

In addition to the fact that GGT activity is elevated in cirrhosis of the liver, acute viral hepatitis, chronic hepatitis, pancreatitis, myocardial infarction, the existence of an obstruction (obstacle) in the bile ducts, primary or metastatic liver tumors, decompensated heart disease and malignant pancreatic tumor, as well as liver damage caused by alcoholism, it is also closely related to the use of hepatotoxic drugs.

Any change in GGT in subjects selected according to the mentioned criteria would without any doubt indicate the hepatotoxicity of the tested product which is the subject of the patent.

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Bearing in mind that during the application of the product there was a significant drop of 13% and not an increase in the level of GGT activity, it can be concluded that the product in question does not have a hepatotoxic effect on users.

On the other hand, creatine kinase (CK) or creatine phosphokinase (CPK) is an enzyme that catalyzes the reaction of creatine and adenosine triphosphate (ATP), resulting in creatine phosphate and adenosine diphosphate (ADP).

In tissues that quickly consume energy, such as striated muscles, but also the brain, or smooth muscles, phosphocreatine serves as a quick source for ATP regeneration, so creatine kinase is a very important enzyme for that tissue. Normal values for men are < 177 u/L and for women < 153 u/L.

Creatine kinase in the blood in elevated concentrations can be a marker of myocardial infarction, rhabdomyolysis, muscular dystrophy or acute kidney failure. As such, in this study, it is used as an indicator of exactly those side effects of statins, but also of natural monacolins, i.e. monacolin K.

Bearing in mind that during the application of the product there was no increase in CK beyond the permitted values, it can be concluded that the product in question does not pose a risk for the occurrence of common side effects of natural monacolins such as rhabdomyolysis, muscular dystrophy or acute kidney failure.

According to the test results from Table 4, in addition to the CK remaining within the limits of the permitted values, a 13% drop in Gamma GT was recorded, which makes the product non-toxic and safe for use. Also, without a drop in HDL, there was a lowering of the T.DT level. by 28%, as well as total cholesterol by 20%, while triglycerides were reduced by 23%, and the average drop in sugar for all 27 subjects was 2%.

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Example 6. - Changes in cholesterol values

Table 5. - Change in cholesterol level in all 27 subjects

The results of the cholesterol level test at the beginning and at the end of the test, shown in Table 5, which are presented in Figure 1, show that in all 27 subjects, after 45 days of use, there was a drop in blood cholesterol levels by an average of 20%, and that the highest drop was recorded in the subjects with the highest cholesterol level at the beginning of the test. The effect of the preparation is proportional to the initial cholesterol values.

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Example 7 - Blood parameters of subjects (7) with cholesterol < 6 mmol/L at the beginning of the tests a

Table 6 - Blood parameters of subjects with cholesterol < 6 mmol/L The test results given in Table 6 showed that, in the group of subjects who had cholesterol < 6 mmol/L at the beginning of the study, the subject product did not lead to an increase in creatine kinase beyond normal values, although a 20% jump was recorded and there was a 15% decrease in GGT. Also, with an increase in HDL levels by 9.5%, there was a decrease in LDL levels by 23% and total cholesterol by 17%, while triglycerides were reduced by 31%, and the average sugar drop for these 7 subjects was 0.5%.

Example 8 - Blood parameters of the subject (11) with cholesterol > 6 mmol/L at the beginning of the test

Table 7 - Blood parameters of subjects with cholesterol > 6 mmol/L The test results given in Table 7 showed that, in the group of subjects who had cholesterol > 6 mmol/L at the beginning of the study, the product in question did not lead to an increase in creatine kinase beyond normal values, although a jump of 40% was recorded and there was a decrease of GGT by 15%. Also, with an increase in HDL levels by 3%, there was a decrease in LDL levels by 31% and total cholesterol by 16%, while triglycerides were reduced by 21%, and the average sugar drop for these 11 subjects was 3%.

Example 9 - Parameters of subjects (7) with cholesterol > 7 and < 8 mmol/L at the beginning of the study

Table 8. - Blood parameters of subjects (7) with cholesterol > 7 and < 8 mmol/L The test results given in Table 8 showed that, in the group of subjects who at the beginning of the 5 study had cholesterol > 7 and < 8 mmol/L, the subject product did not lead to an increase in creatine kinase outside of normal values, although a jump of 5% was recorded and there was a decrease of GGT by 13%. Also, with an increase in the level of HDL by 0.8%, there was a decrease in the level of LDL by 35% and total cholesterol by 25%, while triglycerides were also reduced by 25%, and the average drop in sugar for these 7 subjects was 5%.

Example 10 - Blood parameters of the subject (2) with cholesterol > 8 mmol/L at the beginning of the study

Table 9. - Blood parameters of subjects (2) with cholesterol > 8 mmol/L The test results given in Table 9 showed that, in the group of subjects who at the beginning of the study had cholesterol > 8 mmol/L, the product in question did not lead to an increase but to a drop of creatine kinase by 38%, which made the CK level remain significantly below normal values and the GGT level remained the same as it was, significantly below the permitted values. Also, with an increase in HDL levels by 21%, there was a decrease in LDL levels by 36% and total cholesterol by 32%, while triglycerides were also reduced by 36%, and there was an average jump in sugar levels for these 2 subjects of 6%.

Example 11. - Changes in triglyceride values

Table 10. - Change in triglyceride levels in all 27 subjects

The results of the triglyceride level test at the beginning and at the end of the test are shown in Table 10.

10, which are presented in Figure 2, show that in all 27 subjects, after 45 days of use, the level of triglycerides in the blood decreased by 17%, and that the largest decrease was recorded in the subjects with the highest cholesterol level at the beginning of the study. The effect of the preparation is proportional to the initial values of triglycerides.

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Example 12 - Change in triglycerides in all n = 27 subjects compared to initial values

Table 11 - Change in triglyceride levels during the test

The listed results from Table 11 show that, using the product that is the subject of this invention, there was an average reduction of triglycerides in the blood of 23%, with the largest loss of 29% recorded in the group of subjects who had the highest level of triglycerides at the beginning of the test (> 2.5 mmol/L), then in the middle group (> 1.5 and < 2.5 mmol/L) the drop was 25%, while in the group of subjects with < 1.5 mmol/L triglycerides the drop was 6%.

The stated test results, which showed that there is only a rare occurrence of mild cramps, flatulence, gas and nausea, which cannot be related to the use of the preparation, with a tendency to decrease during the course of the test, with improved emptying, and especially due to giving a feeling of satiety, confirmed that the formulation that is the subject of the invention can be used as a meal replacement.

With these examples, the product has demonstrated high efficiency in lowering cholesterol and triglyceride levels and a mostly mild effect on reducing blood sugar levels.

The effect of the preparation is proportional to the initial values of cholesterol and triglycerides.

On average, for 45 days of using the preparation, the drop in triglycerides is 17% and cholesterol is 20%, while the drop in LDL was an average of 27%.

During the test, the subjects lost an average of 1.04 kg of body weight, but also 1.22 kg of body fat.

Not a single subject left the trial due to side effects.

The level of GGT was reduced by an average of 8%, and none of the toxicity indicators, neither CK nor GGT, exceeded the permitted values.

All the presented examples illustrate the fact that the formulation of the preparation as described in this patent has successfully solved the technical problem of obtaining and using a natural preparation for lowering blood fats, which contains an extract of fermented red rice containing a mixture of natural statins, of which the content of active monacolin K is at least 0.5% in combination with konjac mannan, without known toxic and side effects.

Claims (10)

PATENT CLAIMS 1. A synergistic pharmaceutical formulation, characterized in that it contains up to 50% by weight of purified glucomannans and up to 20% by weight of a highly purified dry extract of fermented red rice containing a mixture of natural monacolins, with a maximum of 50 ppb of citrinin. 2. Synergistic pharmaceutical formulation according to claim 1, characterized in that it is a purified dry extract of red fermented rice obtained by fermentation with Monascus purpureus so that it contains a minimum of 0.5% by weight of monacolin K. 3. Synergistic pharmaceutical formulation according to claims 1 and 2, characterized in that glucomannan can be konjac glucomannan. 4. Synergistic pharmaceutical formulation according to claims 1 - 3, characterized in that it is in powder form suitable for formulating oral pharmaceutical dosage forms. 5. Synergistic pharmaceutical formulation according to claims 1 - 4, characterized in that it is suitable for formulating a unit dose for oral administration containing 5 mg of monacolin K. 6. Synergistic pharmaceutical formulation according to claims 1 - 5, indicated by the fact that other active substances and excipients can be added to the active substances. 7. Use of a pharmaceutical formulation according to claims 1-6, characterized in that it is used as a meal replacement. 8. A synergistic pharmaceutical formulation according to claims 1 - 6, indicated by the fact that it is used in a daily dose of 10 mg of monacolin K, acts on the suppression of cholesterol synthesis and supports the rapid excretion of cholesterol, blocks re-absorption of cholesterol and metabolic disorders, without adverse toxic and hepatotoxic effects, without increasing the liver enzyme gamma glutamyl transferase (GGT), without increasing creatine kinase (CK), and without the risk of rhabdomyolysis, muscular dystrophy and/or acute kidney failure. 9. Synergistic pharmaceutical formulation according to claims 1 - 6, characterized by the fact that it is used to lower the level of total lipids in the blood serum, lower LDL cholesterol, lower triglycerides, weight loss, acts on atherosclerosis, blood pressure, prevention and treatment of heart and blood flow diseases and other diseases caused by metabolic disorders. 10. The procedure for the preparation of a synergistic pharmaceutical formulation according to claims 1 - 6, characterized by the fact that highly purified dry extract of fermented red rice, with a maximum of 50 ppb citrinin, is added to the highly purified konjac glucomannan, without detected sulfur dioxide and arsenic, in an amount that ensures a content per dose of monacolin K of at least 5 mg, and a mixture of excipients containing 5 - 50% glucose is added monohydrate as a sweetener, 5 - 35% acidity regulator, sodium hydrogencarbonate and citric acid, aroma, 1 - 5% magnesium oxide anti-caking agent and steviol glycoside as an additional sweetener. 2 Published and printed by: Institute for Intellectual Property, Belgrade, Kneginje Ljubice 5 2