Patent Application
20130011500
1. Field of the Invention
The present invention relates to a ginger extract and a medication comprising the ginger extract, more particularly, to a ginger extract which can inhibit the fat-storage function of adipocytes and a medication thereof.
2. Description of the Related Art
Adipose tissue refers to a loose connective tissue composed of adipocytes and is generally divided into two types, white adipose tissue and brown adipose tissue, wherein the white adipose tissue is mainly function as a store of energy and the brown adipose tissue is for thermogenesis. In humans, adipose tissue is primary located in subcutaneous layers, in bone marrow and around internals, not only for reservation of liquid but also providing protective padding for organs.
In grown-up people, there are approximately thirty billions of adipocytes in bodies. The adipocytes playing a key role in store of triglyceride and energy-modulation and however, excess adipocytes in human will lead to serious medical disorders, such as obesity.
Conventional anti-obesity medicine includes sibutramine, also known as meridian; orlistat, also known as xenical; chitonsan; and laxatives, for example magnesium oxide. Meridian will prolong the time effect of norepinephrine and serotonin, accelerate the metabolism of organism and finally to advance the consumption of fat. Xenical will inactivate lipase in small intestine, to inhibit the lipolytic effects of small intestine, as well as the absorption of fatty acid whereof. Chitonsan will provide satiety to user. Yet, laxatives will directly lead to diarrhea, eliminating excess fat from excretes in an enforced approach.
However, the conventional anti-obesity medicine are less effective sometimes and usually accompany with plenty of side effects, such as thirsty, insomnia, heat ache and constipation, which may result in various inconvenience to people. Therefore, there is a pressing need of providing an anti-obesity medication being natural, effective and less risky, for the sake of providing a new strategy for losing weight in an easy and convenient process without any side effects.
The primary objective of this invention is to provide a ginger extract for inhibiting the fat-storage function of adipocytes, which can interference with the fat-storage function of adipocytes so as to improve the issue of obesity.
The secondary objective of this invention is to provide a medication comprising the said ginger extract, and which is obtained from natural plants and is capable of improving obesity due to the natural medical properties of ginger.
A ginger extract for inhibiting the fat-storage function of adipocytes is obtained by a process comprising steps of drying, by providing roots or stems of Alpinia galangal and Zingiber zerumbet, followed by drying the roots or stems of Alpinia galangal and Zingiber zerumbet till water content of the roots or stems of Alpinia galangal and Zingiber zerumbet is lower than 10%, to obtain dry Alpinia galangal and dry Zingiber zerumbet; extracting, by preparing a ginger component by mixing 25% to 75% of the dry Alpinia galangal and 25˜75% of the dry Zingiber zerumbet and then extracting the ginger component with a solvent to obtain a liquid extract; and condensation, by condensing the liquid extract to obtain a ginger extract being capable of inhibiting the fat-storage function of adipocytes.
A medication for inhibiting the fat-storage function of adipocytes, comprises a ginger extract as defined in claim 1; and a medical acceptable excipient.
Further scope of the applicability of the present invention will become apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferable embodiments of the invention, are given by way of illustration only, since various others will become apparent from this detailed description to those skilled in the art.
The present invention will become more fully understood from the detailed description given herein below and the accompanying drawings which are given by way of illustration only, and thus are not limitative of the present invention, and wherein:
PHOTO 1 is a histosection datum of white adipose tissue in HD rats of groups D0-4;
PHOTO 2 is a histosection datum of white adipose tissue in HD rats of groups D1-4;
PHOTO 3 is a histosection datum of white adipose tissue in HD rats of groups D2-4;
PHOTO 4 is a histosection datum of white adipose tissue in HD rats of groups D3-4.
All figures are drawn for ease of explaining the basic teachings of the present invention only; the extensions of the figures with respect to number, position, relationship, and dimensions of the parts to form the preferred embodiment will be explained or will be within the skill of the art after the following teachings of the present invention have been read and understood. Further, the exact dimensions and dimensional proportions conforming to specific force, weight, strength, and similar requirements will likewise be within the skill of the art after the following teachings of the present invention have been read and understood.
With reference to
In the step of “drying S1,” roots or stems of Alpinia galangal and Zingiber zerumbe are prepared and dried individually till the water content of the roots or stems of Alpinia galangal and Zingiber zerumbe decrease lower than 10%, in order to obtain dry Alpinia galangal and dry Zingiber zerumbe. More precisely, in the step of “drying S1,” roots or stems of Alpinia galangal and Zingiber zerumbe is dried but not limit to via a lyophilization, spray drying, evaporation or heating drying.
Alpinia galangal of the present invention, also known as Languas galangal, is a kind of herbal medicine and has medical properties in anti-inflammatory, worm infestation and hypersentivity reaction. Also, Zingiber zerumbe of the present invention, also known as the Shampoo Ginger, is regularly used as food flavor or herbal medicine in various cuisines, and which show medical properties in anti-proliferation of cancer cell, and hypersentivity reaction.
In the step of “extracting S2,” 25% to 75% of the dry Alpinia galangal and 25˜75% of the dry Zingiber zerumbet are mixed up with each other to obtain a ginger component, and then the ginger component is soaked and extracted by a solvent to obtain a liquid extract, wherein the weight ratio between the ginger component and the solvent is 1:8 to 1:12. In the present invention, the solvent is but not limit to water, methanol ethanol acetone, ethane, propane, butane or hexane. In the present embodiment, a ginger component is extracted by 95% ethanol under a process of sonication for 8 hours, for the sake of removing impurities in Alpinia galangal and Zingiber zerumbet, and obtaining the liquid extract of the present invention. With such performance, active substances in Alpinia galangal and Zingiber zerumbet is sufficient to be obtained from the liquid extract, and therefore the liquid extract of the present invention is potential to be put in use in pharmaceutical industry, by manufacturing the liquid extract into any medicament or health products for inhibiting the fat-storage function of adipocytes.
In the step of “condensation S3,” the liquid extract is condensed to obtain a ginger extract for inhibiting the fat-storage function of adipocytes. As an example, the condensation of the liquid extract can be performed by evaporation or heating drying. In the present embodiment, the liquid extract is condensed via a process of evaporation, in order to remove solvent from the liquid extract and to obtain the ginger extract of the present invention. With such condensation, the ginger extract comprising high concentration of active substances of Alpinia galangal and Zingiber zerumbet is successfully obtained, and which will be more significant in use of pharmaceutical industries.
In the next paragraphs, the benefits of the ginger extract in anti-obesity are demonstrated by providing ginger components comprising dry Alpinia galangal and dry Zingiber zerumbet in various weight ratios, and carrying out a serial of trials with those ginger components in the present invention.
Referring to TABLE 1, seven ginger components comprising dry Alpinia galangal and dry Zingiber zerumbet in various weight ratios are prepared and extracted by 95% ethanol to obtain various extracts, and which includes groups (1) comprising dry Alpinia galangal only; (2) comprising dry Zingiber zerumbet only; (3) comprising dry Alpinia galangal and dry Zingiber zerumbet in a weight ratio of 1:1; (4) comprising dry Alpinia galangal and dry Zingiber zerumbet in a weight ratio of 2:1; (5) comprising dry Alpinia galangal and dry Zingiber zerumbet in a weight ratio of 3:1; (6) comprising dry Alpinia galangal and dry Zingiber zerumbet in a weight ratio of 1:2; and (7) comprising dry Alpinia galangal and dry Zingiber zerumbet in a weight ratio of 1:3. More precisely, the total weight of ginger component of groups (1) to (7) is 6 kilograms, and the ginger component of groups (1) to (7) is extracted by 60 to 65 liters of 95% ethanol. In the present embodiment, extracts obtained from groups (1) to (7) are analysis in trials of (A) toxicity, (B) anti fat-storage, and (C) animal.
Trial (A): Toxicity
With reference to TABLE 2, the toxicity of the extracts of groups (1) to (7) are analyzed, by co-incubating a strain of adipocytes, named 3T3-L1 (with a sort code of BCRC 60159) and purchased from Bioresource Collection and Research Center of Food Industry Research and Development Institute in Taiwan, with the ginger extracts of groups (1) to (7) in various dosages, such as 0, 31.25, 62.5, 125, 250, 500 μg/ml, at 37° C., 5% CO2, for 24 hours, and than analyzing the survival rate of the adipocytes 3T3-L1 in each groups via MTT assay. Precisely, 1×104 cells/ml of adipocytes 3T3-L1 are used in each group. In addition, the adipocytes 3T3-L1 is cultured at 37° C., 5% CO2, for 24 hours before the trial (A) of the present embodiment.
In MTT assay of the present invention, the adipocytes 3T3-L1 of each group are co-incubated with MTT reagent (3-(4,5-cimethlthiazol-2-yl)2,5,-diphenl tetrazolium bromide) for a period, and then the optical density (OD) of each groups is analyzed by spectrophotometer. With such arrangement, the tetrazolisum of the MTT (in yellow) will be transferred to formazan (in blue) under the reduction of dehdrogenase in live cells, and therefore, the survival rate of adipocytes 3T3-L1 in each group can be obtained by measuring the OD value the adipocytes 3T3-L1 of each groups under 550 nm.
According to TABLE 3, the survival rate of each group is summarized. It is noted that the survival of groups 1b to 1f, 2b to 2f, 3b to 3f, 4b to 4f, 5b to 5f, 6b to 6f and 7b to 7f are all higher than 90% in comparison with control groups individually, including 1a, 2a, 3a, 4a, 5a, 6a, and 7a, with the survival rate being defined as 100% in the present embodiment. Hence, it is suggested that the ginger extract will not damage to cells.
Trial (B): Anti Fat-Storage
In the trial (B), the anti fat-storage function of ginger extract in the present invention is demonstrated by using oil-red stain. With reference to TABLE 4, the adipocytes 3T3-L1 are prepared and cultured at a DMEM medium contained 10% bovine serum at first, followed by suspending the adipocytes 3T3-L1 with a trypsin-EDTA and counting the cell numbers of adipocytes. Next, 1×106 cells/ml of adipocytes 3T3-L1 in each group are collected and co-incubated with medium I at 37° C., 5% CO2, for 2 days, then transferred to medium II and cultured for 3 days, and finally transferred to medium III for further culturing for 7 to 15 days. In the present embodiment, the adipocytes 3T3-L1 of each group are switched to fresh medium III every two days during the culturing of medium III.
In TABLE 5, it is shown that a control group, having untreated adipocytes 3T3-L1, and the adipocytes 3T3-L1 obtained from the above culturing are further cultured at 37° C., 5% CO2, for 24 hours, and then the adipocytes 3T3-L1 obtained from the above culturing are randomly assigned into 42 groups and co-incubate with ginger extracts of groups (1) to (7) in a various dosage, such as 0, 31.25, 62.5, 125, 250, 500 μg/ml, at 37° C., 5% CO2, for 24 hours. Finally, adipoctes 3T3-L1 in the control group and groups (1A) to (7F) are analyzed via the oil-red stain.
In the present embodiment, the oil-red stain is processed by removing the culturing medium from each group, washing with PBS buffer, keeping at a fix solution, being PBS buffer with 10% formalin, for 1 hour, removing the fix solution, and keeping at 0.2 ml of oil-red dye for another 1 hour, wherein the oil-red dye is prepared by dissolving 50 ml oil-red in 10 ml isoacetone. As following, the processed adipocytes 3T3-L1 of each group are analyzed by spectrophotometer at 510 nm.
In TABLE 6, it is pointed out that the groups of (1A) to (7F) shows significant performance in fat-storage in comparison with the untreated adipocytes 3T3-L1 (C). Furthermore, an anti fat-storage function is observed in groups (1B) to (1F), (2B) to (2F), (3B) to (3F), (4B) to (4F), (5B) to (5F), (6B) to (6F) and (7B) to (7F) in comparison with groups of (1A), (2A), (3A), (4A), (5A), (6A), and (7A) individually, especially in a dose-dependent manner. It is verified that the ginger extract of the present invention is sufficient to inhibit the fat-storage function of adipocytes, particularly to the ginger extract obtained from the ginger component having dry Alpinia galangal and dry Zingiber zerumbe in a weight ratio of 1:3.
Trial (C): animal test
In the present embodiment, male, around 200 to 250 grams and 8-weeks-old Wistar rats purchased from National Laboratory Animal Center in Taiwan are prepared, housed at a standard laboratory environment, such as keeping 25±1° C. and with a 12 hours light/dark cycle, and fed with high fat diet (60 kcal %, TestDiet® Formula 58Y1) for 4 weeks. With such arrangement, high-fat diet-induced obesity rats (HD rats), gaining more than 40% weights, are obtained in the present invention, and which will be used as an animal module in the trial (C) to further demonstrate the anti fat-storage function of the ginger extracts.
With reference to TABLE 7, the HD rats of the present invention are randomly assigned into 16 groups, including (D1), as a control and feeding with 1 ml water during the trial (C); (D2), feeding with ginger extract, containing dry Alpinia galangal and dry Zingiber zerumbe in a weight ratio of 1:3, in a dosage of 150 mg/per kilogram of weight daily during the trial (C); (D3) feeding with ginger extract, containing dry Alpinia galangal and dry Zingiber zerumbe in a weight ratio of 1:3, in a dosage of 300 mg/per kilogram of weight daily during the trial (C); and (D4); feeding with ginger extract, containing dry Alpinia galangal and dry Zingiber zerumbe in a weight ratio of 1:3, in a dosage of 600 mg/per kilogram of weight daily during the trial (C). In the present embodiment, the weights and the weight gain of the HD rats in each group are monitored at the 0, 4, 8, and 12 weeks of the trial (C). Moreover, after the trial (C), white adipose tissues of the HD rats in each group are collected and analyzed by histological staining. In the present embodiment, the white adipose tissues of the HD rats are collected around the epididymis of the HD rats, followed by fixing with 10% formalin, embedding with paraffin, slicing and staining by hematoxylin and eosin stain.
ap < 0.05
In TABLEs 7 and 8, it is indicated that the HD rats has a great amount of weight gain after having high-fat diet, and however, the feeding with ginger extract of the present invention, particularly in a dosage of 300 to 600 mg, will significantly reduce the weights of HD rats (see data in D2 and D3). It is proved that the ginger extract has benefits on anti fat-storage of adipocytes. Additionally, in PHOTOs 1 to 4, histosection photos of white adipose tissues of the HD rats in groups D1 to D4, being 200 times of magnification, are shown. It is suggested that, HD rats fed with the ginger extract of the present invention has smaller but more adipocytes in comparison with control rats (D1). In TABLE 10, numbers of adipocytes of the HD rats in group (D1) to (D4) are measured and recorded at the 12th weeks in trial (C), and it is suggested that feeding with the ginger extract of the present invention will dramatically reduce the size of adipocyte in rats. As a result of reducing the size of adipocytes, the number of the adipocytes in a unit area in the histosection photo is increased. It is believed that the ginger extract of the present invention will significantly recovery the symptoms caused by high-fat diets, such as gaining weights, and increase of adipocyte in size.
It is suggested that the ginger extract of the present invention is potential to be applied to pharmaceutical industry, being an active substance of medication or health products for inhibiting fat-storage function in adipose tissues. In the present invention, the ginger extract can be given to any target individually or combined with any acceptable excipients, for example carriers or other ingredients, and is capable of being further manufactured into any form of medicament, such as pill, capsule, powder, solution and pastil for easy and convenient delivery to targets. Preferably, the medication of the present invention comprises dry Alpinia galangal and dry Zingiber zerumbet in a weight ratio of 1: 3 to 3:1, particularly to 1:3, and is preferably delivered to target once a day, with a dosage of 300 to 600 mg per kilogram of body weight and with a period of treatment lasting for 12 weeks.
In summary, through the present invention, a manufacture method of a ginger extract for inhibiting fat-storage function of adipose tissues is provide, by obtaining a ginger extract comprising dry Alpinia galangal and dry Zingiber zerumbet in a weight ratio of 1:3 to 3:1 via an easy, convenient and time-and-cost saving process. With the ginger extract of the present invention, a medication or health products comprising the ginger extract are also easily obtained, and which has natural medical properties in anti-obesity, and will be easy to put to used in pharmaceutical industries. In this way, the general public can successfully lose weight in an easier, healthier and more convenient process.
Thus, since the invention disclosed herein may be embodied in other specific forms without departing from the spirit or general characteristics thereof, some of which forms have been indicated, the embodiments described herein are to be considered in all respects illustrative and not restrictive. The scope of the invention is to be indicated by the appended claims, rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are intended to be embraced therein.
