WO2007058480A1 - Composition having effect on treatment and prevention of diseases syndrome treatment with glabridin - Google Patents

Abstract

Provided is a composition for prevention and/or treatment of a disease syndrome, comprising glabridin and/or derivatives thereof represented by Formula I in the specification, as an active ingredient, wherein the glabridin and derivatives thereof have a pharmacological activity on prevention and/or treatment of the disease syndrome and exhibit a superior activity on AMP-activated protein kinase (AMPK), and the disease syndrome includes obesity, diabetes, diabetic complications, fatty liver, hypertension, dyslipidemia, arteriosclerosis, cardiovascular diseases, mitochondrial dysfunction-related diseases, and degenerative diseases.

Description

COMPOSITION HAVING EFFECT ON TREATMENT AND PREVENTION OF DISEASES SYNDROME TREATMENT WITH GLABRIDIN

FIELD OF THE INVENTION

The present invention relates to a composition for preventing and treating a disease syndrome, containing glabridin as an active ingredient. More specifically, the present invention relates to a composition for preventing and treating a disease syndrome exhibiting a superior activity on AMP-activated protein kinase (AMPK), i.e. various diseases arising from energy excess such as obesity, diabetes, diabetic complications, fatty liver, hypertension, dyslipidemia, arteriosclerosis, cardiovascular diseases, mitochondrial dysfunction-related diseases, and degenerative diseases, by inclusion of glabridin and/or derivatives thereof as an active ingredient.

BACKGROUND OF THE INVENTION

Recently, improvement in hygienic conditions owing to economic development and hence a rise in the standard of living, and frequent ingestion of instant foods and changes of dietary habits due to Western influence involving consumption of large amounts of meat lead to excessive calorie intake in the body. However, such changes of dietary habits in modern people bring about a rapid increase of overweight and obese population, due to excessive lack of physical exercise and thereby low calorie ^•expenditure.

Therefore, calories excessively accumulated in the body becomes an underlying cause of various diseases including obesity, as well as various diseases such as diabetes, metabolic diseases, degenerative diseases and mitochondrial dysfunction- related diseases.

Obesity, a condition in which an amount of body fat is abnormally high compared to standard body weight, refers to a disease resulting from accumulation of surplus calories in adipose tissues of the body when calorie intake is greater than calorie expenditure. Obesity raises problems associated with the negative body image and low physical appearance of people, and chronicity thereof is reported to cause a variety of adult diseases such as hypertension, diabetes, hyperlipidemia and coronary artery diseases, and cancers such as breast cancer, uterine cancer and colon cancer. Therefore, obesity is regarded as a fatal disease (J. Biol. Chem., 273, 32487 - 32490 (1998); and Nature, 404, 652 - 660 (2000)).

Specifically, complications caused from obesity include, for example hypertension, myocardiac infarction, varicosis, pulmonary embolism, coronary artery diseases, cerebral hemorrhage, senile dementia, Parkinson's disease, type 2 diabetes, hyperlipidemia, cerebral apoplexy, various cancers (such as uterine cancer, breast cancer, prostate cancer, colon cancer and the like), heart diseases, gall bladder diseases, sleep apnea syndrome, arthritis, infertility, venous ulcer, sudden death, fatty liver, hypertrophic cardiomyopathy (HCM), thromboembolism, esophagitis, abdominal wall hernia (Ventral Hernia), urinary incontinence, cardiovascular diseases, endocrine diseases and the like (Obesity Research Vol. 12(8), 2004, 1197-1211).

Diabetes is a systemic metabolic disorder resulting from multiple environmental and genetic factors, and refers to a condition characterized by abnormally elevated blood glucose levels due to absolute or relative deficiency of insulin in the body. Complications of diabetes includes, for example hypoglycemia, ketoacidosis, hyperosmolar coma, macrovascular complications, diabetic retinopathy, diabetic neuropathy, diabetic nephropathy and the like.

Metabolic syndromes refer to syndromes accompanied by health risk factors such as hypertriglyceridemia, hypertension, glycometabolism disorders, blood coagulation disorders and obesity. According to the ATP III criteria of the National

Cholesterol Education Program (NCEP) published in 2001, individuals are diagnosed with the metabolic syndrome by the presence of three or more of the following components: 1) A waistline of 40 inches (102 cm) or more for men and 35 inches (88 cm) or more for women (central obesity as measured by waist circumference), 2) A triglyceride level above 150 mg/dL, 3) A high density lipoprotein (HDL) level less than

40 mg/dL (men) or under 50 mg/dL (women), 4) A blood pressure of 130/85 mm Hg or higher and 5) A fasting blood glucose level greater than 110 mg/dL.

Insulin resistance refers to a phenomenon wherein, even though insulin is normally secreted in the body, "supply of glucose into cells" performed by insulin does not work properly. Therefore, glucose in the blood cannot enter cells, thus causing hyperglycemia, and further, cells themselves cannot perform normal functions thereof due to a shortage of glucose, leading to the manifestation of metabolic syndrome.

The degeneration is the term derived from pathological findings, thus meaning the condition which is accompanied by "decreases in consumption of oxygen", and refers to a degenerative disease wherein dysfunction of mitochondria, which is an organelle that generates energy using oxygen within the cell, is associated with the senescence. As examples of the degenerative disease, mention may be made of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and

Huntington's disease (Korean Society of Medical Biochemistry and Molecular Biology News, 2004, 11(2), 16-22).

Diseases arising from mitochondrial dysfunction may include for example, mitochondrial swelling due to mitochondrial membrane potential malfunction, functional disorders due to oxidative stress such as by the action of active oxygen species or free radicals, functional disorders due to genetic factors, and diseases due to functional deficiency of oxidative phosphorylation mechanisms for energy production of mitochondria. Specific examples of diseases, developed by the above-mentioned pathological causes, may include multiple sclerosis, encephalomyelitis, cerebral radiculitis, peripheral neuropathy, Reye's syndrome, Friedrich's ataxia, Alpers syndrome, MELAS, migraine, psychosis, depression, seizure and dementia, paralytic episode, optic atrophy, optic neuropathy, retinitis pigmentosa, cataract, hyperaldosteronemia, hypoparathyroidism, myopathy, amyotrophy, myoglobinuria, hypotonia, myalgia, the decrease of exercise tolerance, renal tubulopathy, renal failure, hepatic failure, liver function failure, hepatomegaly, red blood cell anemia (iron- deficiency anemia), neutropenia, thrombocytopenia, diarrhea, villous atrophy, multiple vomiting, dysphagia, constipation, sensorineural hearing loss (SNHL), epilepsy, mental retardation, Alzheimer's disease, Parkinson's disease and Huntington's disease (see, for example US Patent No. 6,183,948; Korean Patent Laid-open Publication No. 2004- 7005109; Journal of clinical investigation 111, 303-312, 2003; Mitochondria 74, 1188- 1199, 2003; and Biochimica et Biophysica acta 1658 (2004) 80-88).

The above-mentioned obesity, diabetes, metabolic syndromes, degenerative diseases and mitochondrial dysfunction-related diseases will be collectively referred to as "disease syndrome" hereinafter. Therefore, the term "disease syndrome" used herein refers to a variety of diseases arising from excessive accumulation of surplus calories in the body.

At present, the most effective way to ameliorate or fight against the conditions associated with such disease syndromes is known to be exercising more and losing weight, and dietary control. All of the currently effective ways of fighting against the disease syndrome have in common the fact that they facilitate energy metabolism, thus resulting in maximized expenditure of surplus energy in the body leading to prevention of energy accumulation. Effective expenditure of such surplus energy is considered a method for treating the disease syndrome. Enhancing of a metabolic activity is essential for effective elimination of surplus energy. For this purpose, it is essential to achieve inhibition of lipogenesis, inhibition of gluconeogenesis, facilitation of glucose consumption, facilitation of fat oxidation, facilitation of biogenesis of mitochondria which is a central apparatus of energy metabolism and collective activation of factors involved in metabolic activation. There is yet little known about targets to treat the disease syndrome, whereas numerous target proteins or genes are known only for treating individual diseases and therefore there have been proposed some methods for the prevention or treatment of such diseases via use of the above-mentioned corresponding target proteins or genes. However, there is still room for further significant improvement even in treatment of individual diseases such as metabolic syndromes including obesity, diabetes and the like. In spite of the fact that a great deal of studies has been conducted on treatment of diseases, there are yet no drugs available for the treatment of various diseases resulting from excess energy intake and aging.

In this connection, as one of various attempts to treat the disease syndrome, there is research to control the imbalance of energy metabolism and oxidation-reduction state. Most of diseases including obesity, diabetes, metabolic diseases, degenerative diseases and mitochondrial dysfunction-related diseases, i.e., large numbers of diseases including disease syndromes, stem from the imbalance of energy metabolism and oxidation-reduction state. As it is believed that all of diseases arise from energy excess due to excessive amounts of NAD(P)H functioning as a coenzyme of oxidation- reduction reaction, as the most important cause of such diseases, a method of controlling an activity and amount OfNAD(P)H is under investigation.

Further, there are methods of preventing and/or treating a disease syndrome including obesity, by enhancing energy metabolism of muscle tissues taking advantage of AMP-activated protein kinase (AMPK). The muscle tissues, in conjunction with cardiac tissues, are typical tissues utilizing fatty acids as an energy source, and are crucial for energy metabolism in vivo. Hence, numerous attempts have been made to develop various drugs which are capable of suppressing or preventing obesity by increasing an energy metabolic rate of the muscle tissues. In particular, with recent publication of the fact that AMP-activated protein kinase (AMPK) plays a central role in metabolism of carbohydrates and fat in the muscle tissues, AMPK is recognized as a target material for inhibition of obesity via regulation of energy metabolism.

Meanwhile, the factors and expression phenomena, which are regulated in the downstream mechanism via the activation of AMPK, are provided as follows.

L Glvcometabolism

In muscle tissues and myocardial tissues, AMPK promotes muscle contraction and thereby facilitates intake of glucose. That is, AMPK activates GLUT 1, or induces migration of GLUT 4 to a plasma membrane, irrespective of insulin action, resulting in increased transport of glucose into cells (Arch. Biochem. Biophys. 380, 347-352, 2000,

J. Appl. Physiol. 91, 1073-1083, 2001). After increasing glucose uptake into cells,

AMPK activates hexokinase, thereby increasing flux of glycometabolism processes and simultaneously inhibiting glycogen synthesis. It is known that in myocardial tissues under ischemic conditions, AMPK phosphorylates to activate 6-phosphofructo-2-kinase

(PFK-2), with consequent activation of a metabolic cascade leading to increased flux of glycometabolism (Curr. Biol. 10, 1247-1255, 2000). In addition, the activation of

AMPK in the liver inhibits the release of glucose from hepatocytes. Meanwhile, it was confirmed that the activity of phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase, which are gluconeogenesis enzymes, is inhibited by AMPK

(Diabetes 49, 896-903, 2000), thus indicating that AMPK independently takes part in the control of a blood glucose level via inhibition of the release of glucose from the liver, irrespective of insulin.

2. Mitochondrial Biogenesis

One important function of mitochondria is to carry out an oxidative phosphorylation process, which converts energy produced from fuel metabolites such as glucose and fatty acids into ATP. It is known that the incidence of disorders in mitochondrial functions is involved in a pathogenic mechanism of various degenerative diseases associated with the senescence, such as diabetes, cardiovascular diseases, Parkinson's disease and senile dementia (Curr. Opin. Cell Biol. 15, 706-716, 2003). Peterson et al (Science 300, 1140-1142, 2003) have suggested the possibility that deteriorated mitochondrial function is a probable pathogenic cause of insulin resistance syndrome, with reporting that oxidative phosphorylation functions of mitochondria were weakened by about 40% in the elderly. In addition, they have confirmed that a decrease in the content of mitochondrial DNA in the peripheral blood is appeared from before the incidence of diabetes (Diabetes Res. Clin. Pract. 42, 161-167, 1998). Biogenesis of mitochondria in muscles is known to be promoted by an adaptive reaction in which metabolic activity of oxidative phosphorylation of muscle cells is increased by chronic energy depletion and exercise. Zong et al (Proc Natl. Acad. Sci. USA 99: 15983-15987, 2002) have revealed that, using a transgenic mouse in which AMPK was genetically inactivated, AMPK is required for mitochondrial biogenesis in skeletal muscle cells under conditions in which chronic energy deprivation is induced. Further, Putman et al (J. Physiol. 551, 169-178, 2003) have demonstrated the hypothesis that AMPK in association with continuous exercise is involved in an increase of mitochondrial volume.

Meanwhile, it was confirmed that AMPK increases gene expression of a peroxisome proliferator-activated receptor gamma coactivator lα (PGC- lα) which is known to play an important role in mitochondrial biogenesis (Endocr. Rev. 24, 78-90,

2003). Raynald et al (Am. J. Physiol. Endocrinol. Metab. 281, 1340, 2001) have reported that chronic activation of AMPK plays an important role in activation of a nuclear respiratory factor- 1 (NRF-I) and mitochondrial biogenesis. NRF-I, which is a gene essential for transcription of proteins associated with a mitochondrial respiratory system as well as mitochondrial transcription and replication, plays an important role to increase oxidation capability in muscle cells in response to chronic energy stress.

Therefore, NRF-I consequently participates in an increase of mitochondrial biogenesis.

In addition, it is known that an enzymatic activity of citrate synthase and 3- hydroxyacyl-CoA dehydrogenase, known as being increased in conjunction with

increased amounts of E2-EPF ubiquitin carrier protein-3 (UCP-3) and mRNA thereof and an increased mitochondrial volume, is increased by activation of AMPK (J.

Physiol. 551, 169-178, 2003).

3. Regulation of fat metabolism

Upon reviewing a mechanism of AMPK participating in fat metabolism, AMPK induces phosphorylation of acetyl-CoA carboxylase, thereby resulting in inhibition of fatty acid synthesis. Therefore, AMPK is known to facilitate fatty acid oxidation, by the action of decreasing an intracellular concentration of malonyl-CoA that is an intermediate of fatty acid synthesis and is an inhibitor of carnitine palmitoyl transferase I (CPT I). CPT I is an enzyme essential for a fatty acid oxidation process wherein fatty acids enter mitochondria and are oxidized, and it is known that CPT I is under the control of an intracellular malonyl-CoA concentration. In addition, AMPK is known to inhibit activity of HMG-CoA reductase and glycerol phosphate acyl transferase (GPAT), involved in synthesis of cholesterol and triacylglycerol, through phosphorylation (J. Biol. Chem. 277, 32571-32577, 2002, J. Appl. Physiol. 92, 2475- 2482, 2002). Meanwhile, it was found that activation of AMPK in the liver inhibits the activity of pyruvate kinase, fatty acid synthase and ACC through phosphorylation of carbohydrate-response-element-binding protein (ChREBP) (J. Biol. Chem. 277, 3829- 3835, 2002). In addition, the activity of sterol-regulatory-element binding protein- 1 (SREBP-I), which plays an important role in differentiation of adipocytes, is also inhibited by the action of AMPK, which consequently results in inhibition of adipocyte differentiation.

4, Control of protein synthesis

In the protein synthesis process, AMPK inhibits synthesis of proteins via inhibition of mTOR and p70S6K by activating a tuberous sclerosis complex (TSC), or

AMPK inhibits translation elongation via activation of elongation factor-2 (eEF2) kinase and inactivation of eEF2 through phosphorylation thereof. It was found that eEF2 kinase is a direct substrate for AMPK (J. Biol. Chem. 278, 41970-41976, 2003). As discussed above, a variety of metabolism-related activators are known to play a central role in energy metabolism of glucose, proteins and fat acids in vitro and in vivo. With the study from Kahn's group (Nature, 2002), it was reported that AMPK maintains the body energy balance and will be developed as a promising and noticeable novel drug for treatment of obesity and diabetes, in the form of an enzyme-activating drug. Further, recently in 2004, the Kahn's Group has revealed that AMPK maintains the body energy balance by the hypothalamic control of appetite. Neil et al (Nature drug discovery, 3(April), 340, 2004) has asserted that AMPK and Malonyl-CoA are possible targets for the treatment of metabolic syndromes, and they have also stated that patients suffering from metabolic syndromes can be characterized by insulin resistance, obesity, hypertension, dyslipidemia, and dysfunction of pancreatic beta cells, type 2 diabetes and manifestation of arteriosclerosis. It was hypothesized that a common feature linking these multiple abnormalities is dysregulation of AMPK/Malonyl-CoA energy level- sensing and signaling network. It was proposed that such dysregulation leads to alterations in cellular fatty-acid metabolism that in turn cause abnormal fat accumulation, cellular dysfunction and ultimately disease. Evidence is also presented that factors activating AMPK and/or reducing malonyl-CoA levels might reverse these abnormalities and syndromes or prevent incidence of these diseases.

In addition, Roger et al (Cell, 117, 145-151, 2004) have suggested that AMPK may be a possible target for obesity control by lowering the activity of hypothalamic AMPK, thus increasing a content of malonyl-CoA and consequently controlling appetite for food intake, and they have also suggested that alpha-lipoic acid can exert anti- obesity effects by suppressing hypothalamic AMPK activity, thus controlling appetite (Nature medicine, 13 (June), 2004). Meanwhile, they have also reported that alpha- lipoic acid promotes fat metabolism via activation of AMPK in muscle tissues, not in the hypothalamus, and alpha-lipoic acid is therapeutically effective for the treatment of obesity because it facilitates energy expenditure by activating UCP-I, particularly in adipocytes.

Diraison et al (Diabetes 53, S84-91, 2004) have reported that activation of

AMPK in pancreatic cells leads to four-fold increases in expression of the gut hormone peptide YY responsible for appetite control and thus appetite can also be controlled by the action of AMPK even in other tissues other than the hypothalamus.

Nandakumar et al (Progress in lipid research 42, 238-256, 2003) have proposed that, in ischemic heart diseases, AMPK can be served as a target to treat ischemia reperfusion injuries via regulation of fat and glucose metabolism. Min et al (Am. J.

Physiol. Gastrointest Liver Physiol 287, Gl -6, 2004) have reported that AMPK is effective for the control of alcoholic fatty liver.

Genevieve et al (J. Biol. Chem. 279, 20767-74, 2004) have reported that activation of AMPK inhibits activity of an iNOS enzyme that is an inflammatory mediator in chronic inflammatory conditions including obesity-related diabetic diseases or in endotoxin shock, and thus AMPK will be effective for developing new medicines having a mechanism capable of enhancing insulin sensitivity. In addition, they have reported that inhibition of iNOS activity is effected by activation of AMPK, and thus this finding is clinically applicable to diseases such as septicemia, multiple sclerosis, myocardial infarction, inflammatory bowel diseases and pancreatic beta-cell dysfunction. Zing-ping et al (FEBS Letters 443, 285-289, 1999) have reported that AMPK activates endothelial NO synthase through phosphorylation, in the presence of Ca-calmodulin in murine muscle cells and myocardial cells. This represents that AMPK is implicated in heart diseases including angina pectoris.

Lubna et al (AM. J. Physiol. Cell. Physiol. 286, C1410-1416, 2004) have reported that a myocyte enhancer factor 2 (MEF2) is a transcription factor playing a critical role in differentiation of skeletal muscle cells and is activated in muscle cells by AMPK.

Javier et al (Genes & Develop. 2004) have reported that a lifespan can be extended by limiting utilization of energy, and AMPK participates in lengthening of the lifespan, in a manner that an in vivo AMP/ ATP ratio is increased and the α2 subunit of AMPK is then activated by AMP. Therefore, they have suggested that AMPK may function as a sensor to detect the relationship between lifespan extension and energy level and insulin-like signal information.

As obesity-related metabolism regulated by the activation of AMPK, AMPK is known to play a central role in control of the energy-related metabolism such as inhibition of adipocyte differentiation, regulation of fatty acid synthesis-related enzymes and genes (Acetyl-CoA Carboxylase I and II), activation of mitochondrial biogenesis (PGC- lα), activation of thermogenic genes (UCP-2 and 3), inhibition of diacylglycerol acyltransferase (DGAT) activity and the like.

Further, according to recent and continuous publications of study and research results, it is reported through the molecular biological study that the activation of AMPK is therapeutically effective for the treatment of simple obesity as well as the treatment of heart diseases such as diabetes, hypertension and hyperlipidemia, and obesity-related diseases such as fatty liver and inflammation, in conjunction with a result showing an anti-aging activity such as enhancement of physical strength and endurance and anti-oxidation mechanism.

The present inventors carried out an extensive search for AMPK-activating agents, based on the assumption that materials activating AMPK will be therapeutically effective for treatment of disease syndromes, and as a result, have confirmed that glabridin and derivatives thereof are active ingredients for treatment of the disease syndromes.

SUMMARY OF THE INVENTION

Therefore, the present invention has been made to solve the above problems and other technical problems that have yet to be resolved.

As a result of a variety of extensive and intensive studies and experiments based on the facts as described above, the inventors of the present invention have confirmed that activation of muscle AMPK via the use of a composition of glabridin and its derivatives is effective for prevention and/or treatment of disease syndromes including obesity and diabetes arising from excessive accumulation of surplus calories in the body. The present invention has been completed based on these findings.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS In accordance with an aspect of the present invention, the above and other objects can be accomplished by the provision of a pharmaceutical composition for treatment and/or prevention of a disease syndrome, comprising, as an active ingredient, at least one material selected from the group consisting of glabridin and derivatives thereof, represented by Formula I below:

wherein R and R₁ are each independently hydrogen or an alkyl.

The term "disease syndrome" used herein refers to a variety of diseases arising from excessive accumulation of surplus calories in the body, and may include, for example obesity, diabetes, diabetic complications, fatty liver, hypertension, dyslipidemia, arteriosclerosis, cardiovascular diseases, mitochondrial dysfunction- related diseases, degenerative diseases, and decreases in motor ability and physical endurance.

Glabridin or derivatives thereof, which are utilized as the active ingredient in the composition of the present invention, may be preferably extracted from Licorice (Gan-Cao or sweet herb), a medicinal herb, as main raw materials. Species of Licorice plants include European Licorice (Glycyrrhiza glabra),

Manchurian Licorice (G. uralensis), Russian Licorice (G. glabra var. glandulifera) and

G. pallidiflora. Roots and integuments (with or without skin) of other members belonging to the same family (Leguminosae) may also be used as sources of Licorice plants.

Examples of the glabridin derivatives may include glabrene, Hispaglabridin A, Hispaglabridin B, 4'-0-methylglabridin, 2'-0-methylglabridin, 2',4'-0-methylglabridin and 3'-hydroxy-4'-0-methylglabridin. These ingredients may be contained alone or as a mixture thereof in the composition of the present invention.

Licorice containing the glabridin is primarily known to be effective for anti- oxidative action, whitening action, anti-inflammatory action and the like. Physiological activities of Licorice known hitherto are as follows.

Licorice has various activities such as antagonism against a variety of poisonous or toxic materials, thereby treating medicinal poisoning due to use of powerful drugs or poisonous drugs, and neutralizing and detoxifying action against bacterial toxins, thus exerting neutralizing and alleviating action to abate toxicity of all kinds of drugs, and is therefore recognized as indispensable in Oriental Herbal medicine. In addition, Licorice is also known to have hormone-like regulatory functions similar to adrenocortical hormones, thereby relieving acute and urgent conditions, alleviation of pain arising from muscle tension and relaxation of neurotensin, an antiallergic activity by glycyrrhizin which is the active principle of licorice root, and therapeutic effects on gastric ulcer and duodenal ulcer. Minepei Kuroda et al (Bioorganic & Medicinal Chemistry Letters 13 (2003)

4267-4272) have reported that glycyrin, one of Licorice constituents, serving as a medicinal substance activating PPAR-γ, is effective to control a blood glucose level.

Toshio Fukai et al (Fitoterapia 74 (2003) 624-629) have reported that a glabridin component of Licorice has therapeutic effects on kidney diseases, via the free radical scavenging activity. Ute M. Kent et al (Drug Metabolism and Disposition 30(6) 709-

715, 2002) have reported that the glabridin in Licorice suppresses an activity of human cytochrome P450. However, none of the above-mentioned publications disclosed or suggested prevention and treatment of the disease syndromes via enhancement of an AMPK activity, as demonstrated in the present invention.

The present inventors have newly found that glabridin and derivatives thereof activate an AMPK enzyme playing a central role in metabolism of carbohydrates and fat in the muscle tissues and are thereby capable of preventing and treating the disease syndromes including obesity. These findings and facts can also be demonstrated through the following examples. Specifically, the present inventors have measured effects of glabridin having a physiological activity on the activity of the AMPK enzyme in myoblast cells (C2C12), and as a result, have confirmed that the AMPK enzyme exhibits an excellent activity.

As an example of the disease syndrome, obesity can be treated by a manner that glabridin promotes fat oxidation via activation of AMPK, thereby reducing an amount of surplus energy accumulated in the body and thus treating obesity. The present inventors have confirmed such a fact that the AMPK enzyme exhibits an excellent activity in experiments of administrating glabridin to ob/ob mice, an animal model of obesity/diabetes having loss of an appetite-controlling ability due to deficiency of the leptin gene, and therefore AMPK has therapeutic and prophylactic effects on obesity of mice.

As a result, the composition for prevention and/or treatment of the disease syndrome comprising glabridin and/or derivatives thereof as the active ingredient can prevent and treat the disease syndrome through activation of AMPK, and thus it is considered that they can be developed as various therapeutic agents for a variety of diseases associated with the disease syndromes.

The content of the active ingredient, i.e. glabridin or derivatives thereof, may be appropriately determined depending upon the desired applications of the composition (prophylactic, health or therapeutic treatment) and may preferably in the range of 0.1 to 10% by weight, based on the total weight of the composition.

The composition for prevention and/or treatment of a disease syndrome in accordance with the present invention comprises the above glabridin or derivatives thereof as the active ingredient, and may be formulated into the disease syndrome- prophylactic and therapeutic agent, in conjunction with addition of a pharmaceutically acceptable carrier, if necessary.

A suitable dose of the pharmaceutical composition of the present invention may vary depending upon various factors such as formulation method, administration fashion, age, weight and sex of patients, pathological conditions, diet, administration time, administration route, excretion rate and sensitivity to response. The pharmaceutical composition of a disease syndrome-prophylactic and therapeutic agent according to the present invention comprises glabridin or derivatives thereof as the active ingredient. The glabridin or derivatives thereof can be administered via oral or parenteral routes upon clinical administration and can be used in general forms of pharmaceutical formulations. That is, the composition in accordance with the present invention may be administered in the form of various oral and parenteral formulations, upon practical clinical administration. When formulating, the formulations are prepared using conventional filling agents, extenders, binding agents, wetting agents, disintegrating agents, diluents such as surfactants, or vehicles. Solid formulations for oral administration include, for example, tablets, pills, powders, granules and capsules, and are prepared by mixing the glabridin or derivatives thereof with one or more vehicles, such as starch, calcium carbonate, sucrose, lactose and gelatin. Lubricating agents such as magnesium stearate and talc may also be used, in addition to simple vehicles. As liquid formulations for oral administration, mention may be made of suspensions, solutions for internal use, emulsions and syrups. In addition to generally used simple diluents such as water and liquid paraffin, the above-mentioned formulations can contain various vehicles, for example wetting agents, sweetening agents, aromatics and preservatives. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations and suppositories. As non-aqueous solvents and suspensions, there may be used propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethylolate, etc. As base materials for suppositories, Witepsol, macrogol, Tween 61, cacao butter, laurin butter, glycerol and gelatin may be used.

Dosage units may contain one-, two-, three- or four-fold amount of individual dose, or 1/2, 1/3 or 1 /4-fold amount of individual dose. Preferably, an individual dose contains an amount of the active drug that is administered one time, and typically corresponds to the total amount administered for one day, or 1/2, 1/3 or 1/4 fold-amount thereof. Although effective doses of the glabridin or derivatives thereof are concentration-dependent, they are preferably in the range of 0.1 to 1,000 mg/kg, more preferably 0.4 to 500 mg/kg and may be administered 1 to 6 times a day. Therefore, the glabridin or derivatives thereof may be administered in the range of preferably 0.1 to 6,000 mg/day/kg bw, more preferably 0.4 to 4200 mg/day/kg bw, particularly preferably 0.4 to 700 mg/day/kg bw, for adults.

In accordance with another aspect of the present invention, there is provided a functional health food composition for preventing and/or treating a disease syndrome, containing glabridin or derivatives thereof as an active ingredient. The term "a functional health food" used throughout the specification of the present invention refers to a food in which glabridin or derivatives thereof are added to general foods to improve functions thereof. The glabridin or derivatives thereof may be added to general foods or may be prepared in the form of capsules, powders, suspensions and the like. Intake of such a functional health food containing glabridin or derivatives thereof provides beneficial effects for health, and exhibits advantages in that there are no adverse side effects caused by prolonged use of drugs because food material is used as the raw material, unlike conventional drugs.

If it is desired to use glabridin or derivatives thereof of the present invention as a food additive, the glabridin or derivatives thereof can be added alone, or can be used in conjunction with other foods or food ingredients, or may be used appropriately according to other conventional methods. Mixed amounts of active ingredients may be suitably determined depending upon the purpose of use. Generally, in producing foods or beverages with which the glabridin or derivatives thereof are mixed, these materials may be added in an amount of 0.0001 to 10% by weight, and preferably in an amount of 0.1 to 5% by weight, based on the total weight of raw materials. However, when prolonged intake is intended for the purpose of health and hygiene or for health control, the above-mentioned amount of the glabridin or derivatives thereof may be adjusted below the above-mentioned range. In addition, the health food of the present invention preferably contains glabridin or derivatives thereof in amounts falling within the determined toxicity range, when it is employed as a pharmaceutical composition.

There is no particular limit to kinds of the above-mentioned foods. As examples of foods to which the glabridin or derivatives thereof can be added, mention may be made of meats, sausages, bread, chocolate, candies, snack, confectionary, pizza, Ramen, other noodles, gum, skimmed milk, dried foods, raw foods, dairy products including lactic acid bacteria-fermented milk and ice cream, various soups, beverages, teas, drinks, alcoholic beverages and multi-vitamin preparations. Specifically, as examples of health foods containing the glabridin or derivatives thereof, mention may be made of health foods and special favorite products such as squeezed liquid, tea, jelly and juice made of the glabridin or derivatives thereof as main ingredients. In addition, mention may be made of folk medicines for edema, nephritis and urethritis as targets.

When it is desired to use the glabridin or derivatives thereof of the present invention as cosmetic raw materials, they can be added by themselves or can be used in conjunction with other cosmetic ingredients, or may be used appropriately according to other conventional methods. Mixed amounts of active ingredients may be suitably determined depending upon the purpose of use thereof. Generally, in producing cosmetics using the glabridin or derivatives thereof, these derivatives may be added in an amount of 0.0001 to 10% by weight, and preferably in the amount of 0.1 to 5% by weight, based on the total weight of raw materials. Cosmetics include, but are not limited to, aftershaves, lotions, creams, packs and color cosmetics.

The glabridin or derivatives thereof according to the present invention may be extracted using Licorice as dried medicinal material or raw medicinal material, or may be synthesized by organochemical methods.

A process for extracting glabridin or derivatives thereof from Licorice comprises: a) subjecting Licorice to water or organic solvent extraction to obtain crude extracts, b) filtering the crude extracts, followed by (vacuum) concentration, and c) optionally, removing the solvent.

Preferred examples of the organic solvent may include lower alcohols such as methanol, ethanol and the like, and acetone.

For example, Licorice is first extracted with acetone, vacuum concentrated and then re-extracted with methylene chloride to obtain a concentrated solution. The thus- obtained solution is purified via silica column chromatography to obtain pure glabridin.

BRIEF DESCRIPTION OF THE DRAWINGS The above and other objects, features and other advantages of the present invention will be more clearly understood from the following detailed description taken in conjunction with the accompanying drawings, in which:

FIG. 1 is a bar graph comparing activity of AMPK (AMP-activated protein kinase) between the treatment group and control group, after treatment of myoblast cell line C2C12 with glabridin;

FIG. 2 is a micrograph showing effects of glabridin on phosphorylation of AMPK and ACC in C57BL/6 mice;

FIG. 3 is a graph comparing changes in body weight of animals, after 30-day administration of glabridin to an animal model of obesity, adult ob/ob mice;

FIG. 4 is a graph showing changes in levels of blood lipids, inflammatory factors and glucose, after administration of glabridin to ob/ob mice;

FIG. 5 is a micrograph showing effects of glabridin on liver tissues of obese (ob/ob) mice;

FIG. 6 is a graph showing effects of glabridin on mitochondrial biogenesis;

FIG. 7 is a micrograph showing increases of mitochondrial number and improvement of mitochondrial function in liver tissues, after administration of glabridin to ob/ob mice; FIG. 8 is a graph showing effects of glabridin on control of blood pressure in animals suffering from hypertensive diseases;

FIG. 9 is a graph showing effects of glabridin on control of a blood glucose level in an animal model of type 1 diabetes;

FIG. 10 is a graph showing effects of glabridin on prevention of a cataract;

FIG. 11 is a micrograph comparing a difference in a severity of a cataract for glabridin-administered group and a control group;

FIG. 12 is a graph showing effects of glabridin on control of a blood glucose level in an animal model of type 2 diabetes;

FIG. 13 is a micrograph of fat-staining showing effects of glabridin on alcoholic fatty liver in an animal model of alcoholic fatty liver;

FIG. 14 is a graph showing changes in levels of lipid-related indicator materials in the blood, after administration of glabridin in a rat model of alcoholic fatty

liver;

FIG. 15 is a graph showing effects of glabridin on spontaneous locomotor and activity of an animal model; and

FIG. 16 is a graph showing effects of glabridin on enhancement of physical endurance of an animal model. EXAMPLES

Now, the present invention will be described in more detail with reference to the following Examples. These examples are provided only for illustrating the present invention and should not be construed as limiting the scope and spirit of the present invention.

Example 1 : Isolation of Glabridin

For recovery of glabridin from Licorice {Glycyrrhiza glabra), partially purified glabridin (Daepyung Co., Ltd. Gyeonggi-Do, KOREA) was purchased and completely purified for use. NMR analysis results of the thus-purified glabridin are as follows:

- NMR and Mass Data for Glabridin

IH-NMR (CDC13, 300.40 MHz): δ 6.95(d, IH), 6.82(d, IH), 6.65(d, IH), 6.38(dd, IH), 6.37(d, IH), 6.3 l(d, IH), 5.56(d, IH), 5.20(b, 2H), 4.37(m, IH), 4.02(t, IH), 3.48(m, IH), 2.97(dd, IH), 2.85(m, IH), 1.43(s, 3H), 1.41(s, 3H).

13C-NMR (CDC13, 75.45 MHz): δ 155.25, 154.44, 151.91, 149.75, 129.18, 128.95, 128.41, 120.01, 116.95, 114.32, 109.93, 108.73, 107.98, 103.11, 75.62, 70.00, 31.70, 30.61, 27.79, 27.55.

Mass (ApCI) m/z 325(M+1).

Example 2: Determination of AMPK activity Myoblast cells, C2C12, were cultured in DMEM containing 10% bovine calf serum. When a cell density reached a range of about 85% to 90%, the culture medium was replaced with a medium containing 1% bovine calf serum to induce differentiation of cells. Enzymatic activity of AMPK was determined as follows. Firstly, C2C12 cells were lysed to obtain protein extracts and then ammonium sulfate was added to a final concentration of 30%, thereby precipitating proteins. Protein precipitates were dissolved in a buffer (62.5 mM Hepes, pH 7.2, 62.5 mM NaCl, 62.5 mM NaF, 1.25 mM Na pyrophosphate, 1.25 mM EDTA, 1 mM DTT, 0.1 mM PMSF, and 200 μM AMP). Thereafter, 200 μM SAMS peptide (HMRSAMSGLHLVKRR: the underlined serine residue is a phosphorylation site, as an AMPK phosphorylation site of acetyl-CoA carboxylase) and [γ-32P]ATP were added thereto and reactants were reacted for 10 minutes at 30 ^°C . This was followed by spotting of the resulting reaction solution on p81 phosphocellulose paper. The p81 paper was washed with a 3% phosphoric acid solution and radioactivity thereof was measured. For each reaction condition, reactions involving no SAMS peptide were also conducted and basic values were subtracted from the thus- observed values.

FIG. 1 is a bar graph comparing activity of AMPK between the treatment group and control group, after treatment of myoblast cell line C2C12 with glabridin. As can be seen from FIG. 1, the treatment of myoblast cell line C2C12 with glabridin resulted in an increased activity of AMPK.

Example 3: Effects of glabridin on phosphorylation of AMPK & ACC (acetyl-CoA carboxylase^') in C57BL/6 mice In order to investigate anti-obesity effects of glabridin, the glabridin was administered daily to obese mice at a dose of 150 mg/kg via an oral route, and effects of glabridin on phosphorylation of AMPK and ACC, which play an crucial role in energy metabolism and lipogenesis in the liver tissues, were examined.

As shown in FIG. 2 illustrating effects of glabridin on phosphorylation of

AMPK and ACC in C57BL/6 mice, it was confirmed through Western blot analysis that the glabridin as an AMPK activator has an effect on phosphorylation of AMPK and ACC in the liver tissues of C57BL/6 mice. Therefore, the phosphorylated AMPK is believed to activate energy-related metabolism. Whereas, it is believed that ACC, which is affected by activation of AMPK, is phosphorylated and lipogenic activity thereof is then inhibited, which will then exert some effects on lipid metabolism, including inhibition of obesity.

Example 4: Assay of prophylactic and therapeutic effects of glabridin on obesity in animal model of obesity, Lep ob/Lep ob mice

C57BL/6JL Lep ob/Lep ob male mice suffer from decreased secretion of leptin due to mutation of the leptin gene and thus continuously and excessively consume feed due to their uncontrolled appetite. As a result, fat is excessively accumulated in the animal body and blood glucose level is elevated, thus reaching a body weight of about 50 g after about 10 to 11 weeks of birth.

In order to examine prophylactic and therapeutic effects of obesity by glabridin, adult Lep ob/Lep ob male mice, weighing about 50g, were divided into three groups, two experimental groups and one control group, consisting of 10 animals each. Each 10 mice of two experimental groups was respectively administered glabridin at a dose of 75 mg/kg/day and 150 mg/kg/day, for 30 consecutive days. Whereas, 10 mice of the control group were administered an equal amount of vehicle alone, instead of glabridin. According to measurement and analysis results of changes in the body weight over administration time between the experimental groups and the control group, as shown in FIG. 3, it was confirmed that the glabridin-administered experimental groups exhibited a significant decrease of the body weight in a dose-dependent manner, as compared to the body weight of the control group.

Example 5: Changes of blood lipid, hepatic inflammatory factor and glucose level by administration of glabridin

10- week-old C57BL/6J ob/ob male mice were housed and allowed to acclimate to a new environment for five days prior to the experiment, in a breeding room maintained at a temperature of 22±2^°C, humidity of 55±5%, and a 12-h light/dark (LfD) cycle (light from 8:00 a.m. to 8:00 p.m.). Animals were fed a solid feed (5053, Labdiet) as a laboratory animal feed.

Animals were divided into two groups, one experimental group with administration of glabridin (200 mg/kg) and one control group. Samples were orally administered. C57BL/6J ob/ob mice, in conjunction with the control group, were orally administered glabridin at a dose of 200 mg/kg for 8 weeks. Sera were collected after 8 weeks, and serum levels of triglyceride, cholesterol, GOT, GPT and glucose were measured. The results thus obtained are shown in FIG. 4. As shown in FIG. 4, the glabridin-administered group exhibited significant decreases in levels of triglyceride, cholesterol, GOT and GPT, as compared to the control group.

Example 6: Effects of glabridin on liver tissues of obese mice

Livers removed from mice of Example 4 were histologically analyzed. For control group mice and glabridin-administered obese mice, distribution of adipose tissues in the liver was confirmed by H&E staining. FIG. 5 shows the results thus obtained.

As shown in FIG. 5, it was confirmed through fat staining of adipose tissues that administration of glabridin resulted in a pronounced reduction of fat accumulation in the liver of the treatment group, as compared to the control group of obese mice.

Therefore, it was confirmed that glabridin can be used for the prevention and/or treatment of fatty liver.

Example 7: Effects of glabridin on biogenesis of mitochondria

In order to confirm increases or decreases in biogenesis of mitochondria, which is a small intracellular organelle responsible for energy production and cellular respiration, after administration of glabridin, levels of PGC lα, NRF-I, mtTFA, CPTl, AMPK alpha 1 & AMPK alpha 2 mRNA were confirmed using real-time quantitative PCR. The results thus obtained are shown in FIG. 6 and 7. As shown in FIG. 6, it was confirmed that levels of genes involved in mitochondrial biogenesis were significantly increased at a late phase in the liver tissues, as compared to the control group.

In addition, as shown in FIG. 7 representing increases of mitochondrial number and improvement of mitochondrial function in liver tissues, the results of EM examination on the liver tissues confirmed remarkably decreased fat vacuoles and glycogen stores as compared to a control group, recovery of normal mitochondrial shape, significant increases in mitochondrial number, and improved shapes of endoplasmic reticulum. Therefore, it is believed that the increases of mitochondrial number and improvement of mitochondrial function will be effective for the treatment of degenerative diseases and genetic diseases arising from mitochondrial dysfunction.

Example 8: Effects of glabridin on hypertension

6-week-old SHR male rats were housed and allowed to acclimate to a new environment for 15 days prior to the experiment, in a breeding room maintained at a temperature of 22±2^°C, humidity of 55±5%, and a 12-h light/dark (LfD) cycle (light from 8:00 a.m. to 8:00 p.m.). Animals were fed a solid feed (5053, Labdiet) as a laboratory animal feed. Animals were divided into three groups, to which a vehicle, 100 mg/kg of glabridin and 150 mg/kg of glabridin are administered, respectively. Samples were orally administered. Blood pressure of animals was measured using a sphygmomanometer (LE5002, LSI LWTICA, Barcelona, Spain) every 7 days. The results thus obtained are shown in FIG. 8. Experiments were carried out for 30 days. As can be seen from FIG. 8, it was confirmed that administration of glabridin leads to significant control of the blood pressure.

Example 9: Effects of glabridin on type 1 diabetes

6-week-old SD male rats were housed and allowed to acclimate to a new environment for 7 days prior to the experiment, in a breeding room maintained at a temperature of 22±2°C, humidity of 55±5%, and a 12-h light/dark (L/D) cycle (light from 8:00 a.m. to 8:00 p.m.). Animals were fed a solid feed (5053, Labdiet) as a laboratory animal feed. Animals having an average body weight of 200 to 220 g were selected and fasted for 6 hours. Animals were administered 30 mg/kg of streptozotocin (STZ, Sigma Co., USA) dissolved in 0.1 M citrate buffer (pH 4.5) via blood-vessel injection such as intravenous injection and were divided into three groups: one control group, one group with administration of a vehicle, and one group with administration of glabridin (150 mg/kg). Samples were orally administered from 10 days after treatment of animals with streptozotocin. Experiments were carried out for 28 days after induction of type 1 diabetes. The results thus obtained are shown in FIG. 9.

As can be seen from FIG. 9, it was confirmed that administration of glabridin leads to significant control of the blood glucose level in an animal model of type 1 diabetes, as compared to the control group.

Example 10: Prophylactic and therapeutic effects of glabridin on cataract

6-week-old SD male rats were housed and allowed to acclimate to a new environment for 7 days prior to the experiment, in a breeding room maintained at a temperature of 22±2^°C, humidity of 55±5%, and a 12-h light/dark (L/D) cycle (light from 8:00 a.m. to 8:00 p.m.). Animals were fed a solid feed (5053, Labdiet) as a laboratory animal feed. Animals having an average body weight of 200 to 220 g were selected and fasted for 6 hours. Animals were administered 30 mg/kg of streptozotocin (STZ, Sigma Co., USA) dissolved in 0.1 M citrate buffer (pH 4.5) via blood-vessel injection such as intravenous injection and were divided into three groups: one control group, one group with administration of a vehicle, and one group with administration of glabridin (150 mg/kg). Samples were orally administered from 10 days after treatment of animals with streptozotocin. Experiments were carried out for 60 days after induction of type 1 diabetes. The severity of cataract was determined using a slit lamp. The severity of cataract was classified on a scale of 0 to 5.

As a result, as shown in FIGS 10 and 11 showing an incidence rate of a cataract and photographs comparing a difference in the severity of a cataract, respectively, it was confirmed that the glabridin-administered group exhibited effective inhibition of cataract induction, as compared to the control group.

Example 11 : Effects of glabridin on type 2 diabetes

18-week-old Zucker Diabetic Fatty (ZDF) male rats (available from Charles River Laboratories, Noblesville, IN, USA), as an animal model of type 2 diabetes, were housed and allowed to acclimate to a new environment for 5 days prior to the experiment, in a breeding room maintained at a temperature of 22±2 ^°C , humidity of 55±5%, and a 12-h light/dark (L/D) cycle (light from 8:00 a.m. to 8:00 p.m.). Animals were fed a solid feed (5053, Labdiet) as a laboratory animal feed. Animals were divided into two groups: one control group and one group with oral administration of glabridin (200 mg/kg). Animals were fed ad libitum an animal feed. Experiments were carried out for 25 days. The results thus obtained are shown in FIG. 12.

As can be seen from FIG. 12, it was confirmed that the glabridin-administered group exhibited effective control of the blood glucose level, as compared to the control group.

Example 12: Therapeutic effects of glabridin on fatty liver

An alcoholic fatty liver model was established by feeding a rat with a mixture of ethanol and drinking water, and improving effects of glabridin on fatty liver was observed. 8 weeks after administration of glabridin, H&E staining and oil-red O staining of the liver tissues were performed. FIG. 13 shows a micrograph of fat- staining and FIG. 14 shows changes in levels of lipid-related indicator materials in the blood.

As shown in FIG. 13, a significant decrease in an amount of lipid droplets was observed after 8-week administration of glabridin, and there was no observation showing signs of pathological abnormal findings in the liver tissues. The lipid-related indicator materials and biochemical indicators in the blood were analyzed after administration of glabridin to the alcoholic fatty liver rat model.

As shown in FIG. 14, the blood level (numerical value) of GOT and GPT as inflammatory indicators, and cholesterol and triglyceride as lipid materials were significantly decreased, as compared to the vehicle-administered group. Example 13: Effects of glabridin on spontaneous locomotor activity

Glabridin was administered to ob/ob C57BL/6 mice, and 3 hours later, spontaneous locomotor activity was measured using Versa MAX Activity Monitors & Analyzer (AccuSan Instruments, Columbus, OH, USA). The monitor used to measure motion of animals was a 41cm x 41cm Plexiglas chamber (height: 30 cm) equipped with infrared rays at intervals of 2.5 cm along the x- and y-axes, respectively, whereby 16 scanning lines are respectively arranged on front/rear and right/left sides of the chamber. In order to distinguish between spontaneous locomotor and stereotypic/grooming behavior, animal activity was measured by taking continuous interference of two different scanning lines caused by mice as an effective determination standard. The glabridin-administered group, the vehicle-administered group and the control group were respectively placed in each measuring apparatus, and activity and motion of animals were measured for 7 hours. For acclimation of animals to a new environment, mice were placed in the apparatus 2 hours prior to measurement. The measurement results thus obtained are shown in FIG. 15.

As shown in FIG. 15, the vehicle-administered group and control group exhibited substantially no difference therebetween, but the glabridin-administered group exhibited a more than 2-fold significant difference in the motion and locomotor activity of animals.

Example 14: Effects of glabridin on enhancement of physical endurance This Example was intended to measure a difference in physical endurance of mice through a swimming test. For this purpose, water was placed in a cylindrical trough having a diameter of 9.5 cm and a height of 25 cm, and glabridin was administered to ob/ob C57BL/6 mice. 3 hours later, a sample-administered group and a control group were placed simultaneously into each cylindrical trough for measurement, and physical endurance of each group was measured and compared. The results thus obtained are shown in FIG. 16.

As shown in FIG. 16, it was confirmed that the glabridin-administered group exhibited about two-fold swimming duration by single administration of glabridin, as compared to the control group.

Example 15: Preparation of Tablet

Glabridin 600 g

Milk serum protein 240 g

Crystalline cellulose 14O g

Magnesium stearate 1O g

Hydroxypropylmethylcellulose 10 g

Example 16: Preparation of Powder

Glabridin 1O g Soybean protein 50 g

Carboxycellulose 40 g

Total 100 g

The above-enumerated components were finely ground and mixed to prepare a powder. 500 mg of the resulting powder was filled into a gelatin hard capsule to prepare a capsule.

Example 17: Application of glabridin to milk

Milk 99.9%

Glabridin 0.1%

Example 18: Application of glabridin to orange juice

Liquid fructose 5%

Polydextrose 1%

Citric acid 5%

Vitamin C 0.02%

Glabridin 0.1% Concentrates of orange fruit juice 25%

Water 67%

Example 19: Preparation of beverage

Calcium lactate 50 ing

Citric acid 5 rag

Nicotinic amide 10 rag

Riboflavin sodium hydrochloride 3 rag

Pyridoxine hydrochloride 2 rag

Arginine 30 mg

Glabridin 100 rag

Water 200 m£

INDUSTRIAL APPLICABILITY

As apparent from the foregoing, glabridin or derivatives thereof according to the present invention can be developed as foods, cosmetics and pharmaceutical compositions for prevention and/or treatment of disease syndromes such as obesity, diabetes, diabetic complications, fatty liver, hypertension, dyslipidemia, arteriosclerosis, cardiovascular diseases, mitochondrial dysfunction-related diseases, degenerative diseases, and decreases in motor ability and physical endurance, via the activation of AMPK.

Although the preferred embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that various modifications, additions and substitutions are possible, without departing from the scope and spirit of the invention as disclosed in the accompanying claims.

Claims

WHAT IS CLAIMED IS

1. A pharmaceutical composition for treatment and/or prevention of a disease syndrome, comprising, as an active ingredient, at least one material selected from the group consisting of glabridin and derivatives thereof, represented by Formula I:

wherein R and R₁ are each independently hydrogen or an alkyl.

2. The composition according to claim 1, wherein the disease syndrome is obesity, diabetes, diabetic complications, fatty liver, hypertension, dyslipidemia, arteriosclerosis, cardiovascular diseases, mitochondrial dysfunction-related diseases, degenerative diseases, or a decrease in motor ability or physical endurance.

3. The composition according to claim 1, wherein the glabridin or derivatives thereof are extracted from Licorice.

4. The composition according to claim 1, wherein the glabridin derivative is selected from the group consisting of glabrene, Hispaglabridin A, Hispaglabridin B, 4'- 0-methylglabridin, 2'-0-methylglabridin, 2',4'-0-methylglabridin, 3'-hydroxy-4'-0- methylglabridin and any combination thereof.

5. The composition according to claim 1, wherein the content of the active ingredient is in the range of 0.1 to 10% by weight, based on the total weight of the composition.

6. The composition according to claim 1, wherein the composition further includes a pharmaceutically acceptable vehicle and/or carrier.

7. The composition according to claim 1 , wherein the composition includes one or more cosmetically or nutritionally acceptable carriers and/or vehicles.

8. A pharmaceutical preparation comprising the pharmaceutical composition of any one of claims 1 to 6.

9. The preparation according to claim 8, wherein the preparation is prepared in the form of a solid formulation for oral administration including a tablet, a pill, a powder, a granule and a capsule, a liquid formulation for oral administration including a suspension, a solution for internal use, an emulsion and a syrup, a formulation for parenteral administration including a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized formulation and a suppository.

10. The preparation according to claim 8, wherein the active ingredient of the preparation is administered in the range of 0.1 to 6,000 mg/day/kg bw, for adults.

11. A process for preparing Licorice extracts containing a material of Formula I of claim 1, comprising: a) subjecting Licorice to water or organic solvent extraction to obtain crude extracts,

b) filtering the crude extracts, followed by (vacuum) concentration, and

c) optionally, removing the solvent.

12. The process according to claim 11, wherein Licorice is a dried medicinal material or raw medicinal material.