WO2005072758A1 - Composition comprising hovenia dulcis thunb. extract, lindera obtusiloba blume extract, or herbal mixture extract thereof - Google Patents
Composition comprising hovenia dulcis thunb. extract, lindera obtusiloba blume extract, or herbal mixture extract thereof Download PDFInfo
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- WO2005072758A1 WO2005072758A1 PCT/KR2005/000283 KR2005000283W WO2005072758A1 WO 2005072758 A1 WO2005072758 A1 WO 2005072758A1 KR 2005000283 W KR2005000283 W KR 2005000283W WO 2005072758 A1 WO2005072758 A1 WO 2005072758A1
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- extract
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- hovenia dulcis
- lindera
- dulcis thunb
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/72—Rhamnaceae (Buckthorn family), e.g. buckthorn, chewstick or umbrella-tree
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/54—Lauraceae (Laurel family), e.g. cinnamon or sassafras
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/18—Antioxidants, e.g. antiradicals
Definitions
- the present invention relates to a composition
- a composition comprising Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof as an effective ingredient .
- liver disease is caused by liver damage resulted from chronic fatigue by stress and most of exogeneous substances.
- the development rate of liver disease in Korea is very high, comparing to foreign countries, and in particular, the death rate of liver cancer is the top in the world and the death rate of chronic liver disease is the third.
- liver disease is the leading cause of death of adults at the age of 40s in Korea.
- the most fatal disease is viral hepatitis in Korea.
- the death by liver cirrhosis is 5 ⁇ 10 fold higher than that by viral hepatitis in Western countries .
- Liver is the organ that shows the most active metabolism among many human organs.
- fatty diet and heavy drinking of alcohol cause fatty liver, that is, lipid is accumulated in liver tissues.
- GOT glutamate-oxaloacetate transaminase
- GPT glutamate-pyruvate transaminase
- Y -GTP Y -glutamyl transpeptidase
- liver disease Since liver has great buffer capacity, liver disease hardly shows symptoms in early stage, and only when it gets serious, it shows symptoms.
- Liver has functions of preservation and circulation of blood, regulation of blood flow and detoxification, and is also known to be involved in mental activity. Human bodies are exposed on pollutions and lots of poison according to industrialization, so that the liver has to overwork in order to counteract poison. Liver damage by mental stress is worse. If human mind gets rest, damaged liver cells are recovered. The thing is that there is no extra time for rest for modern citizens. So, excessive mental stress, heavy smoke and alcohol drinking make liver damage worse, resulting in mal-function of detoxification, that is mal-function of immune system, which might be a cause of another disease.
- Liver cirrhosis is caused by fibrosis of liver tissues, which is the condition of losing homeostasis between generation and degeneration of connective tissue. That is, connective tissue is over-accumulated in liver tissues, accompanying necrosis or inflammation. Fibrosis of liver tissues is progressed by the excessive generation of connective tissue resulted from the growth and transfer of hepatic stellate cells (HSCs) transformed in the form of myofibroblasts (Gressner et al . , Biochem . Biophys. Res . Commun . 151-222, 1988) .
- HSCs hepatic stellate cells
- Liver cirrhosis is the final stage of every chronic liver diseases, by which liver loses its general functions rapidly with showing the decrease of hepatic blood flow, the reduction of blood flow in liver, mal-function of metabolizing enzyme, qualitative and quantitative changes in blood protein, variation of bile flux, etc.
- Hepatotoxicity inducers are exemplified by carbon tetrachloride (CC1 4 ), D-galactosamine, lipopolysaccharide (LPS) and bromobenzene, and in particular, carbon tetrachloride is known to induce hepatotoxicity by inducting lipid peroxidation of liver cells (Recknagel et al., Pharmacol . Rev.
- Carbon tetrachloride is a hepatotoxicity inducer that has been used to investigate anti-hepatotoxic activity. Precisely, it has been administered to mice or rats, cultured liver cells and cultured liver tissues to induce hepatotoxicity artificially. Carbon tetrachloride is converted into the highly reactive molecular structure of free radical CC1 3 • in endoplasmic reticulum by metabolizing enzyme like cytochrome P450, by which it acquires a very strong hepatotoxic effect.
- Free radical CC1 3 • oxidizes triglyceride accumulated in liver by alcohol and fatty acid on membrane phospholipid, leading to acidification and oxidation of lipid, followed by lipid peroxidation. As a result, organic peroxide is produced. Through such lipid peroxidation, fat is accumulated in liver, protein synthesis is reduced, glycogen is decomposed, and cytoplasmic enzymes in blood vessel are destroyed, resulting in necrosis of liver tissues (Chang I, M., et al . , Drug and chemical toxicology 6, 443-453, 1983), Free radical also harms Golgi apparatus, so that protein release out of cells is affected, suggesting that not only liver but also kidney might be damaged.
- Anti-oxidation not only inhibits or prevents lipid peroxidation but also has liver protective and anti- inflammatory effects.
- an antioxidant compound has been used for the protection of liver and liver cells against attacks by reactive oxygen intermediate.
- Antioxidant agents separated from natural resources such as flavonoid, quercetin, silymarin or vitamin E have been reported to have positive effect on lipid peroxidation and hepatic fibrosis.
- N-acetylcysteine (NAC) has been confirmed to reduce oxidative stress and hepatic fibrosis in the early stage of hepatic fibrosis by its anti-oxidative activity.
- Picrorhiza Kurroa (kutkin) is another natural anti-oxidative compound that has liver protective and anti-fibrosis effects by reducing damage caused by lipid peroxidation and free radical.
- the treatment methods for liver diseases are largely divided into two ways; one is a dietetic therapy and the other is medicinal therapy. In most cases, the two methods are used together.
- the two methods are used together.
- liver cell reproduction stimulator and/or liver function protector such as ursodeoxycholic acid, silymarin ⁇ Biotech . Therapeutics, 4, 263-270, 1993), DDB (biphenyl dimethyl dicarboxylate; Biochem . Biophy. Res . Comm .
- liver disease is developed not by just one reason but by co-work of many factors. Thus, one medicine having a certain function is not enough for the treatment of every kinds of liver disease.
- the conventional medicine for the treatment of liver diseases has problems of unexpected reaction and other side effects by long-term or mass administration.
- kidney has functions of regulating the concentrations of ion and body fluids, urinating waste products (urea, uric acid, creatinine, etc) and eliminating toxic substances, drugs and metabolites out of the system after counteracting poison.
- Malonedialdehyde (MDA) and 4-hydroxynonenal (HNE) products from lipid peroxidation in tissues, have been known as indices for cell damage.
- superoxide dismutase catalizing lipid peroxidation has been known to be involved in recovery from cell damage (Slater TF. Biochem. J.
- alkaline phosphatase (ALP) and blood urea nitrogen (BUN) are also in use as diagnostic indices for kidney functions.
- ALP alkaline phosphatase
- BUN blood urea nitrogen
- GFR glomerular filtration rate
- Prerenal azotemia means kidney mal-function without kidney damage, which is resulted from hypoperfusion by congestive heart failure, shock, hypovolemia and hemorrhage.
- postrenal azotemia is caused when the flow of urin is blocked in the lower part of kidney.
- uremia In the case of postrenal azotemia, full recovery is expected by the elimination of the cause. When azotemia accompanies various clinical symptoms and biochemical disorders, it is called uremia. Thus, uremia is not only a simple biochemical disorder but also a syndrome carrying metabolic disorders, endocrine disorders, gastrointestinal disorders, neuromuscular disorders and cardiovascular disorders, in addition to elimination disorders. Diseases caused by mal-function of kidney are exemplified by acute pyelonephritis, chronic pyelonephritis, renal tuberculosis, UTI, urinary stone, renal cell cancer, etc. Hovenia dulcis Thunb.
- Lindera obstiloba is a kind of deciduous broad- leaved tall tree belonging to Lauraceae, which is distributed widely in Korea, Japan, China and Manchuria.
- the flowers, leaves and stems of Lindera obstiloba emit unique fragrance .
- the stems of it have been used for medicinal purposes, owing to their anti-bacterial effect. Fresh shoots of it have been used for tea.
- the present invention provides a compound comprising Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof as an effective ingredient.
- the composition of the present invention includes an anti-oxidative composition, an anti-fibrosis composition, a liver function improving composition and a kidney function improving composition.
- the extracting method for Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention is as follows. Hovenia dulcis Thunb or Lindera obstiloba is washed with water and then dried in the shadow. The dried Hovenia dulcis Thunb or Lindera obstiloba is put in a reflex extractor, to which purified water is added and heated at 100°C for 90 minutes, leading to hot water extraction. The hot water extract is filtered with a filter paper under reduced pressure when it is still hot. The filtrate is concentrated by using a vacuum evaporator. For long-term storage, the solution is dried by using a freeze dryer.
- the stems, flowers, leaves and seeds of Hovenia dulcis Thunb or Lindera obstiloba are all available in the present invention.
- the composition rate of herbs for the composition of the present invention is determined based on the dry weight of each herb. Precisely, Hovenia dulcis Thunb and Lindera obstiloba are mixed at the ratio of 3:2 ⁇ 1:1, and more preferably mixed at the ratio of 2:1 ⁇ 1:1. Such ratio is determined under the consideration of effective dosage of each herb and side effects of it, and the pharmaceutical effects are dropped rapidly or side effects might be a problem when the ratio is out of the above range .
- GOT, GPT, ALP, BUN and total bilirubin which are all liver function indices, are all lowered in the group administered with Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof, indicating that the extract of the present invention has excellent liver function improving effect.
- ALP and BUN are not only used as liver function indices but also used as kidney function indices.
- the lowered levels of ALP and BUN indicate that Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention has excellent kidney function improving effect, as well.
- Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention reduces the amount of hydroxyproline in liver tissues but increases that in kidney tissues, indicating that the extract of the invention has excellent anti- fibrosis and kidney protective effects.
- the herbal extract extracted from the mixture of Hovenia dulcis Thunb and Lindera obstiloba shows higher effects than each individual extract, that is the mixture extract reduces hydroxyproline in liver tissues more but increases it in kidney tissues more than each individual extract.
- Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention can reduce the level of malondialdehyde, an index for lipid peroxidation in liver and kidney tissues, indicating that the extract of the invention has excellent anti-oxidative effect.
- Liver cell line and kidney cell line treated with Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention show high cell viability, indicating that the extract of the present invention has excellent liver cell and kidney cell protective effects. Therefore, the composition of the present invention can be effectively used for the improvement and the protection of liver and kidney functions and for anti- oxidation and anti-fibrosis as well.
- the composition of the present invention can additionally include, in addition to Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof, one or more effective ingredients having the same or similar functions to the extract of the present invention.
- Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention can be administered orally or parenterally and be used in general forms of pharmaceutical formulation.
- the herbal mixture extract of the present invention can be prepared for oral or parenteral administration by mixing with generally used fillers, extenders, binders, wetting agents, disintegrating agents, diluents such as surfactant, or excipients .
- Solid formulations for oral administration are tablets, pills, dusting powders, granules and capsules.
- Solid formulations are prepared by mixing one or more suitable excipients such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. Except for the simple excipients, lubricants, for example magnesium stearate, talc, etc, can be used.
- suitable excipients such as starch, calcium carbonate, sucrose, lactose, gelatin, etc.
- lubricants for example magnesium stearate, talc, etc
- Liquid formulations for oral administration are suspensions, solutions, emulsions and syrups, and the above-mentioned formulations can contain various excipients such as wetting agents, sweeteners, aromatics and preservatives in addition to generally used simple diluents such as water and liquid paraffin.
- Formulations for parenteral administration are sterilized aqueous ' solutions, water-insoluble excipients, suspensions, emulsions, freeze-drying agent and suppositories.
- Water insoluble excipients and suspensions can contain, in addition to the active compound or compounds, propylene glycol, polyethylene glycol, vegetable oil like olive oil, injectable ester like ethylolate, etc.
- Suppositories can contain, in addition to the active compound or compounds, witepsol, acrogol, tween 61, cacao butter, laurin butter, glycerol, gelatin, etc.
- the dosage units can contain, for example, 1, 2, 3 or 4 individual doses or 1/2, 1/3 or 1/4 of an individual dose.
- An individual dose preferably contains the amount of active compound which is administered in one application and which usually corresponds to a whole, 1/2, 1/3 or 1/4 of a daily dose.
- the effective dosage of Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention is 200 ⁇ 600 mg/kg, respectively, and more preferably 300 ⁇ 400 mg/kg, and the administration times are 1 ⁇ 6 times a day.
- the composition of the present invention can be administered singly or combination with surgical operation, radiotherapy, hormone therapy, chemotherapy and biological reaction regulator, to improve liver and kidney functions and to obtain anti-oxidative and anti-fibrosis effects.
- the composition of the present invention can be added in health food to improve liver and kidney functions and to obtain anti-oxidative and anti-fibrosis effects.
- Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention can be added as it is or after being mixed with other food or ingredients, according to the conventional method.
- the mixing ratio of effective ingredients is determined by the purpose of use (prevention, health or therapeutic treatment) .
- the extract is preferably added by under 15 weight%, more preferably under 10 weight%, to the raw material.
- the content of the extract might be less than the above when it is administered for long-term to improve health conditions but the effective dosage could contain more than the above amount because the extract of the invention is very safe.
- Health beverages comprising the composition of the present invention can additionally include various flavors or natural carbohydrates, etc, like other beverages.
- the natural carbohydrates above can be one of monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xilytole, sorbitol and erythritol.
- the ratio of natural carbohydrate to the composition of the present invention is preferably 0.01-0.04 g to 100 mfc, more preferably 0.02-0.03 g to 100 mi .
- the composition of the present invention can include in variety of nutrients, vitamines, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, arginic acid and its salts, organic acid, protective colloidal viscosifiers , pH regulators, stabilizers, antiseptics, glycerin, alcohols, carbonators which used to be added to soda, etc.
- composition of the present invention can also include natural fruit juice, fruit beverages and/or fruit flesh addable to vegetable beverages. All the mentioned ingredients can be added singly or together. The mixing ratio of those ingredients does not matter in fact, but in general, each can be added by 0.01-0.1 weight part per 100 weight part of the composition of the invention.
- Example 1 Preparation of Hovenia dulcis Thunb extract Hovenia dulcis Thumb was washed with water and then dried in the shadow. 20 g of the dried Hovenia dulcis Thunb was put in a reflex extractor, to which 1.5 I of purified water was added and was heated at 100 ° C for 90 minutes. The hot water extract was filtered with a filter paper under the reduced pressure when it was still hot. The filtrate was concentrated by using a vacuum evaporator. It was dried by a freeze dryer for long-term storage. The concentrated solution was used for animal test (2 m£/rat/day) .
- Test animal 12 week old Sprague-Dawley rats weighing about 180-210 g were used as test animals. Breeding temperature was 23 ⁇ 2 ° C and relative humidity was maintained as 60+10%. Feed (Purina feed) and water were freely given, and day-night cycle was regulated. The animals were adapted to the test room for 2 weeks, and then divided into 5 groups; ⁇ normal group, ⁇ CC1 4 treating group, CC1 4 + herbal mixture extract treating group, ⁇ CC1 4 + Hovenia dulcis Thunb extract treating group and ® CC1 4 + Lindera obstiloba extract treating group. Each group was composed of 10 rats.
- obstiloba extract treating group was administered orally with Lindera obstiloba extract prepared in the above example 2 also by 2 mft/rat/day. Rats in every group including the normal group were weighed, and then anesthetized by ether. Blood was taken from heart by heart puncture, which was left at room temperature for over 2 hours. Then, centrifugation was performed at 3000 rpm for 10 minutes to obtain serum, which was stored at -20°C. The serum was examined to measure the level of GOT, GPT, alkaline phosphatase, BUN and total bilirubin. Liver and kidney of a rat, which were already damaged by artificial inducement, were extracted and washed with phosphate buffer (pH 7.0), followed by measuring the weight.
- phosphate buffer pH 7.0
- liver and kidney tissues were kept at -75 °C for the use of measuring hydroxyproline and malondialdehyde (MDA) . Weight changes were measured every week. Ears, tail and feet were investigated to detect the sign of jaundice. The liver was weighed as the rat was sacrificed. The results were presented by mean+SD, and the significance of difference in averages between control group and experimental group was examined by student's t- test. It was judged as statistically significant when p ⁇ 0.005. The results are shown in Table 1.
- GPT (ALT) measurement (using E BIEL kit) 150 f of ALT substrate solution was put in two falcon tubes, which were heated at 37 ° C for 4 minutes. The substrate solution was diluted with standard solution in one tube, and in the other tube, 100 ⁇ i of serum sample was added and reaction was induced at 37 ° C for 30 minutes in both tubes. 100 ⁇ i of purified water and 500 ⁇ Jl of coloring solution (2, 3-dinitrophenylhydrazine) were added to each tube, which was left at room temperature for 20 minutes. 1.5 ml of 0.4 N NaOH was added to each test tube, followed by reaction at room temperature for 10 minutes. OD was measured at 505nm.
- ALP Alkaline phosphatase measurement 2.0 mi of ALP substrate buffer (sodium-2- phenylphosphate) was put in three test tubes, which were reacted at 37 ° C for 3-5 minutes. 50 ⁇ Jl of serum sample, 50 ⁇ Jl of purified water and 50 ⁇ i of standard solution were added to test tubes respectively, followed by reaction at 37 ° C for 15 minutes. 2.0 mi of coloring reagent was added to each test tube and then left at room temperature for 10 minutes. OD was measured at 570nm within 60 minutes.
- ALP substrate buffer sodium-2- phenylphosphate
- BUN Bood urea nitrogen
- ALP and BUN are the indices not only for the liver function but also for the kidney function, so that the lowered levels of ALP and BUN by the extract of the invention also mean that Hovenia dulcis Thunb extract, Lindera obstiloba extract and herbal mixture extract thereof of the present invention have excellent kidney function improving effect as well.
- MDA Malondialdehyde
- Homogenized tissue sample [0.2 g of liver tissues (or kidney tissues) in 1.8 mi of 1.15% KC1] and 200 ⁇ i of diluted standard material (0, 4, 8, 16, 32 nmol/200 ⁇ i tetramethoxypropane) were put together in falcon tube. 200 ⁇ i of standard solution diluted with sample was mixed with 100 ⁇ i of 0.2% SDS, followed by reaction at room temperature for 10 minutes. 750 ⁇ i of 20% acetic acid and 750 ⁇ i of 0.8% thiobarbiturate were added thereto. The solution was reacted at 95 ° C for 30 minutes, then cooled down in ice.
- MTT [3-(4,5-Dimethylthiazol-2-yl) -2 ,5-diphenyl tetrazolium bromide] assay
- 1 mi of trypsin-EDTA solution was added, which was left for 10-20 minutes.
- Cells were isolated from the vessel and then transferred to 15 mi falcon tube.
- 1 mi of medium was added to culture flask and shacked to isolate all the remaining cells, which were also put in the falcon tube. 10 ⁇ l of cell suspension was put in hemocytometer and then cells were counted (2.5X10 4 cell/ml) .
- the solution was cultured in a 37 ° C, 5% C0 2 incubator for 24 hours. At this time, the extract should be added later, otherwise cells are damaged. So, medium should be added first and then the extract was put therein. Upon culturing, the 96 well plate was taken to remove medium. 50 ⁇ i/ml of MTT dye (50 ⁇ l of storage solution + 950 ⁇ l of medium) was added to dilute the solution, which was distributed in each well by 50 ⁇ l, followed by further culture in a C0 2 incubator for 4 hours. After eliminating supernatant, 100 ⁇ i of DMSO was added, followed by stirring for 10 minutes. OD was measured at 540nm with ELISA reader. Only medium was added to a control, and viability was investigated.
- MTT dye 50 ⁇ l of storage solution + 950 ⁇ l of medium
- NR Neuronal Red: 3-amino-7-dimethylamino-2-methyl phenazine assay After removing medium, 1 mi of trypsin-EDTA solution was added, which was left for 10-20 minutes. Cells were isolated from the vessel and then transferred to 15 mi falcon tube. 1 ml of medium was added to culture flask and shaked to isolate all the remaining cells, which were also put in the falcon tube. 10 ⁇ l of cell suspension was put in hemocytometer and then cells were counted (2.5X10 4 cell/ml) . Cells were inoculated to 96 well plate by 50 ⁇ l (1-2X10 5 cells) /well, to which 150 ⁇ l of medium (DMEM + 10% FBS + antibiotics) was added.
- the cells were cultured in a 37 °C, 5% C0 2 incubator for 24 hours, resulting in cell adhesion. 24 hours later, medium was carefully removed from the cells adhered onto the 96 well plate (Careful attention is required for the medium not to be stained with cells and contaminated) .
- 150 ⁇ l of medium was supplemented with 50 ⁇ l of Hovenia dulcis Thunb extract prepared in example 1, 50 ⁇ l of Lindera obstiloba extract prepared in example 2 and 50 ⁇ l of herbal mixture extract prepared in example 3, making total volume 200 ⁇ l .
- the solution was cultured in a 37 "C, 5% C0 2 incubator for 24 hours. At this time, the extract should be added later, otherwise cells are damaged.
- Lindera obstiloba extract and herbal mixture extract of the present invention showed high cell viability in liver cell line.
- herbal mixture extract is more effective to increase cell viability in liver cell line than Hovenia dulcis Thunb extract.
- Hovenia dulcis Thunb extract, Lindera obstiloba extract and herbal mixture extract thereof of the present invention did not harm cell viability in kidney cell line. Therefore, Hovenia dulcis Thunb extract, Lindera obstiloba extract and herbal mixture extract thereof of the present invention were confirmed to have excellent liver and kidney protective effects.
- compositions comprising Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention were prepared as follows.
- Preparative Example 1 Preparation of pharmaceutical compositions 1. Preparation of powders Hovenia dulcis Thunb extract (or Lindera obstiloba extract or herbal mixture extract thereof) 2g Lactose lg The above-mentioned ingredients were mixed together, and airtight bag was filled with the mixture to prepare powders .
- Preparation of food Food comprising Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention was prepared as follows. 1. Preparation of spices and condiments Spices and condiments for health improvement that contains Hovenia dulcis Thunb extract (or Lindera obstiloba extract or herbal mixture extract thereof) of the present invention by 20-95 weight% were prepared.
- Tomato ketchup or source for health improvement was prepared by adding Hovenia dulcis Thunb extract (or Lindera obstiloba extract or herbal mixture extract thereof) of the present invention by 0.2-1.0 weight% to tomato ketchup or source.
- ground beef Health improving ground beef was prepared by adding Hovenia dulcis Thunb extract (or Lindera obstiloba extract or herbal mixture extract thereof) of the present invention by 10 weight% to ground beef.
- Crops (brown rice 30 weight%, job's tears 15 weight%, barley 20 weight%) , Seeds ⁇ Perilla japonica 7 weight%, black bean 8 weight%, black sesame 7 weight%), Dried powder of Hovenia dulcis Thunb extract (or Lindera obstiloba extract or herbal mixture extract thereof) (3 weight%), Ganoderma lucidum (0.5 weight%), Rehmannia glutinosa (0.5 weight%)
- Preparation of carbonated beverage Sugar (5-10%), citric acid (0.05-0.3%), caramel (0.005-0.02%) and vitamin C (0.1-1%) were mixed together, to which purified water (79-94%) was added, resulting in syrup.
- the syrup was sterilized at 85-98 ° C for 20-180 seconds, then mixed with cooling water at the ratio of 1:4.
- Carbon dioxide was injected by 0.5-0.82% thereto, resulting in the preparation of carbonated beverage comprising Hovenia dulcis Thunb extract (or Lindera obstiloba extract or herbal mixture extract thereof) of the present invention.
- Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention is highly effective for the improvement of liver functions since it can lower the level of GOT, GPT, ALP, BUN and total bilirubin, which are major liver function indices.
- ALP and BUN are also used as kidney function indices, so the decrease of the level of ALP and BUN by the extract of the present invention indicates that Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the invention can improve kidney functions as well.
- Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention also can lower the amount of hydroxyproline in liver but increase the amount of hydroxyproline in kidney, suggesting that the extract above has excellent anti- fibrosis and kidney protecting effects.
- Hovenia dulcis Thunb extract, Lindera obstiloba extract or herbal mixture extract thereof of the present invention can lower the level of malondialdehyde, an index of lipid peroxidation in liver and kidney tissues, suggesting that the extract has excellent anti-oxidative effect.
- the composition of the present invention can be effectively used not only for anti-oxidation and anti-fibrosis but also for the protection and the improvement of liver and kidney functions.
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Application Number | Priority Date | Filing Date | Title |
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EP05726304A EP1718167A4 (en) | 2004-01-31 | 2005-01-31 | Composition comprising hovenia dulcis thunb. extract, lindera obtusiloba blume extract, or herbal mixture extract thereof |
JP2006550953A JP2007519715A (en) | 2004-01-31 | 2005-01-31 | A composition comprising a kabonashi extract, a dangwei extract, or a mixed herbal extract thereof as an active ingredient |
CN2005800035902A CN1913910B (en) | 2004-01-31 | 2005-01-31 | Composition comprising hovenia dulcis thunb. extract, lindera obtusiloba blume extract, or herbal mixture extract thereof |
US10/587,286 US20070160699A1 (en) | 2004-01-31 | 2005-01-31 | Composition comprising hovenia dulcis thunb. extract, lindera obtusiloba blume extract, or herbal mixture extract thereof |
US12/465,830 US7846484B2 (en) | 2004-01-31 | 2009-05-14 | Composition comprising Hovenia dulcis thunb. extract, Lindera obtusiloba blume extract, or herbal mixture extract thereof |
Applications Claiming Priority (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR10-2004-0006436 | 2004-01-31 | ||
KR10-2004-0006436A KR100536855B1 (en) | 2004-01-31 | 2004-01-31 | Composition comprising Hovenia dulcis Thunb. extract for improvement of kidney function |
KR10-2004-0006435A KR100536856B1 (en) | 2004-01-31 | 2004-01-31 | Composition comprising herbal mixture extract |
KR10-2004-0006437 | 2004-01-31 | ||
KR1020040006437A KR100586269B1 (en) | 2004-01-31 | 2004-01-31 | Composition comprising Lindera obtusiloba extract |
KR10-2004-0006435 | 2004-01-31 |
Related Child Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US10/587,286 A-371-Of-International US20070160699A1 (en) | 2004-01-31 | 2005-01-31 | Composition comprising hovenia dulcis thunb. extract, lindera obtusiloba blume extract, or herbal mixture extract thereof |
US12/465,830 Continuation US7846484B2 (en) | 2004-01-31 | 2009-05-14 | Composition comprising Hovenia dulcis thunb. extract, Lindera obtusiloba blume extract, or herbal mixture extract thereof |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2005072758A1 true WO2005072758A1 (en) | 2005-08-11 |
Family
ID=34831001
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2005/000283 WO2005072758A1 (en) | 2004-01-31 | 2005-01-31 | Composition comprising hovenia dulcis thunb. extract, lindera obtusiloba blume extract, or herbal mixture extract thereof |
Country Status (4)
Country | Link |
---|---|
US (2) | US20070160699A1 (en) |
EP (1) | EP1718167A4 (en) |
JP (1) | JP2007519715A (en) |
WO (1) | WO2005072758A1 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010082785A2 (en) * | 2009-01-16 | 2010-07-22 | 한올제약주식회사 | Oral pharmaceutical composition |
US20120301562A1 (en) * | 2009-11-20 | 2012-11-29 | Han Wha Pharma Co., Ltd. | Composition for improving blood circulation, containing extract of lindera obtusiloba as active ingredient |
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US9101160B2 (en) | 2005-11-23 | 2015-08-11 | The Coca-Cola Company | Condiments with high-potency sweetener |
US8017168B2 (en) | 2006-11-02 | 2011-09-13 | The Coca-Cola Company | High-potency sweetener composition with rubisco protein, rubiscolin, rubiscolin derivatives, ace inhibitory peptides, and combinations thereof, and compositions sweetened therewith |
KR100989093B1 (en) | 2008-01-18 | 2010-10-25 | 한화제약주식회사 | Composition Comprising the Extracts of Branch of Lindera obtusiloba for Prevention and Treatment of Cardiovascular Diseases |
KR101040682B1 (en) * | 2010-11-04 | 2011-06-10 | 김병주 | The composition where the exemption force is strengthened and the composition water which follows in him |
CN103222995B (en) * | 2013-05-15 | 2014-09-03 | 李伟权 | Application of gynura divaricata in preparing liver-protecting drug or health-care food |
CN111603509A (en) * | 2020-06-30 | 2020-09-01 | 河南中大恒源生物科技股份有限公司 | Preparation method of hovenia dulcis thunb extract rich in dihydromyricetin |
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KR100214801B1 (en) * | 1996-10-05 | 1999-08-02 | 신국현 | Antibiotics isolated from lindera obstiloba |
WO2002060463A1 (en) * | 2001-01-31 | 2002-08-08 | Lifetree Biotech Co., Ltd. | Lower alcohol-insoluble extract of hovenia dulcis var. koreana nakai, a polysaccharide isolated therefrom and an antihepatotoxic composition containing same |
KR20020076980A (en) * | 2001-03-29 | 2002-10-11 | 박근형 | Antiaging Agent Using Water Extracts from the Leaves and Stems of Hovenia dulcis Thunb Containing Natural Antioxidants |
WO2003059369A1 (en) * | 2002-01-17 | 2003-07-24 | Lifetree Biotech Co., Ltd. | Lower alcohol-insoluble extract of hovenia dulcis thunb and a polysaccharide isolated therefrom |
KR100403722B1 (en) * | 2002-05-09 | 2003-11-05 | Forest Genetis Res Inst | Lower alcohol insoluble extract of hovenia dulcis thunberg having liver protection activity, polysaccharide separated therefrom and composition containing the same |
KR100403720B1 (en) * | 2002-01-17 | 2003-11-05 | (주)생명의나무 | Lower alcohol insoluble extract and a polysaccharide therein isolated from the young branches of hovenia dulcis thunb. having antihepatotoxic, anti-hangover and anti-fatigue activity and composition containing same |
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JPS6037999A (en) * | 1983-08-09 | 1985-02-27 | Osaka Chem Lab | Simple method for testing drug effective to hepatopathy |
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JP2003128571A (en) * | 2001-10-22 | 2003-05-08 | Matsuura Yakugyo Kk | Diabetic medicine and health food |
KR20020062898A (en) * | 2002-07-08 | 2002-07-31 | 권만덕 | Lindera obtusitoba bl tea |
KR100543082B1 (en) * | 2003-08-08 | 2006-01-20 | 강대식 | A manufacturing process of composition for reducing the body weight and cholesterol |
-
2005
- 2005-01-31 EP EP05726304A patent/EP1718167A4/en not_active Withdrawn
- 2005-01-31 JP JP2006550953A patent/JP2007519715A/en active Pending
- 2005-01-31 WO PCT/KR2005/000283 patent/WO2005072758A1/en active Application Filing
- 2005-01-31 US US10/587,286 patent/US20070160699A1/en not_active Abandoned
-
2009
- 2009-05-14 US US12/465,830 patent/US7846484B2/en not_active Expired - Fee Related
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KR100214801B1 (en) * | 1996-10-05 | 1999-08-02 | 신국현 | Antibiotics isolated from lindera obstiloba |
WO2002060463A1 (en) * | 2001-01-31 | 2002-08-08 | Lifetree Biotech Co., Ltd. | Lower alcohol-insoluble extract of hovenia dulcis var. koreana nakai, a polysaccharide isolated therefrom and an antihepatotoxic composition containing same |
KR20020076980A (en) * | 2001-03-29 | 2002-10-11 | 박근형 | Antiaging Agent Using Water Extracts from the Leaves and Stems of Hovenia dulcis Thunb Containing Natural Antioxidants |
WO2003059369A1 (en) * | 2002-01-17 | 2003-07-24 | Lifetree Biotech Co., Ltd. | Lower alcohol-insoluble extract of hovenia dulcis thunb and a polysaccharide isolated therefrom |
KR100403720B1 (en) * | 2002-01-17 | 2003-11-05 | (주)생명의나무 | Lower alcohol insoluble extract and a polysaccharide therein isolated from the young branches of hovenia dulcis thunb. having antihepatotoxic, anti-hangover and anti-fatigue activity and composition containing same |
KR100403722B1 (en) * | 2002-05-09 | 2003-11-05 | Forest Genetis Res Inst | Lower alcohol insoluble extract of hovenia dulcis thunberg having liver protection activity, polysaccharide separated therefrom and composition containing the same |
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HASE K. ET AL: "Hepatoprotective effect of Hovenia dulcis THUNB, on experimental liver injuries induced by carbon tetrachloride or D-galactosamine/lipopolysaccharide", BIOL. PHARM. BULL., vol. 20, no. 4, 1997, pages 381 - 385, XP002992805 * |
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YOSHIKAWA M. ET AL: "Bioactive constituents of Chinese natural medicine.III. Absolute stereostructures of new dihydroflavonols, hovenitins I,II,III, isolated from hoveniae semen seufrctus, the seed and fruit of Hovenia dulcis THUNB.(Rhamnaceae): Inhibitory effect on alcohol...", YAKUGAKU ZASSHI, vol. 117, no. 2, 1997, pages 108 - 118, XP001223536 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010082785A2 (en) * | 2009-01-16 | 2010-07-22 | 한올제약주식회사 | Oral pharmaceutical composition |
WO2010082785A3 (en) * | 2009-01-16 | 2010-11-25 | 한올바이오파마주식회사 | Oral pharmaceutical composition |
US20120301562A1 (en) * | 2009-11-20 | 2012-11-29 | Han Wha Pharma Co., Ltd. | Composition for improving blood circulation, containing extract of lindera obtusiloba as active ingredient |
Also Published As
Publication number | Publication date |
---|---|
US7846484B2 (en) | 2010-12-07 |
EP1718167A1 (en) | 2006-11-08 |
US20090285914A1 (en) | 2009-11-19 |
US20070160699A1 (en) | 2007-07-12 |
JP2007519715A (en) | 2007-07-19 |
EP1718167A4 (en) | 2010-07-14 |
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