TWI745609B - Composition for promoting antioxidative activity - Google Patents

Abstract

A composition for promoting antioxidative activity includes an effective dose of Rhodiola extract, a glycerylphosphoryl choline and a pharmaceutically acceptable vehicle or salt thereof. Based on animal experiments, the combination of Rhodiola extract and glycerylphosphoryl choline not only effectively lowers body fat in animal bodies, but also provides high antioxidative activity.

Description

Composition with antioxidant activity

The present invention relates to a composition, especially a composition with antioxidant activity.

Check, human growth hormone (hGH) is a peptide hormone secreted by the pituitary gland. The human growth hormone in the human body has the effect of helping the repair and regeneration of tissue cells, maintaining the activation of various organs, increasing bone density and reducing body fat and cholesterol. However, the secretion of human growth hormone in the human body will decrease year by year after the age of 25. Coupled with the impact of life pressure and environmental deterioration, the body's functions will gradually decline and age.

The above-mentioned problems of human obesity and functional aging can be improved through diet and exercise. If it is not effective, it needs to be treated with drugs. Moreover, many scholars have carried out a number of biochemical experiments in order to achieve the purpose of inhibiting fat formation and delaying aging. Among them, research has found that Rhodiola rosea plant has the effects of anti-anoxia, anti-fatigue, anti-aging and regulating the endocrine system, so many weight loss and anti-aging compounds will add Rhodiola ingredients.

For example, the United States Patent Publication US9737580B2, which discloses a composition for enhancing brain function and a method thereof, the composition contains Huperzine A, Vinpocetine, Acetyl-L-Carnitine and Rhodiola, which is clinically proven, In a specific preferred ratio, it can increase emotional cognition and achieve weight control for Purpose. However, although the ingredients of the above-disclosed composition can effectively regulate body weight, the main purpose is to enhance brain function, thereby enhancing the effects of brain memory and emotional cognition, and it has not been clearly confirmed that it can achieve the purpose of delaying functional aging.

Another example is the Taiwan Invention Announcement Patent No. I441643, which discloses a composition for regulating blood lipids and protecting the cardiovascular system. The composition contains rhodiola powder, red yeast, plant alcohol, natto and vitamin B group, which is confirmed by animal experiments. It can achieve the effect of adjusting blood lipids and protecting the cardiovascular system. Moreover, given a specific proportion of doses, it can significantly reduce body weight; but it has not been clearly proven that it can effectively achieve the antioxidant effect.

The main purpose of the present invention is to solve the problem that the conventional composition cannot improve body fat accumulation and functional aging at the same time.

In order to achieve the above objective, the present invention provides a composition with antioxidant activity. The composition comprises an effective dose of Rhodiola rosea extract and monoglycerophosphocholine and its pharmaceutically acceptable carrier or pharmaceutically acceptable salt.

In this way, through obesity induction and aging experiments, it is confirmed that Rhodiola rosea extract and glycerophosphocholine can effectively reduce the body weight and body fat of animals, and even enhance the antioxidant effect in the animal body, thereby achieving effective weight loss and delaying aging Purpose.

Fig. 1 is an image diagram of hair follicle and subcutaneous fat staining performed on the skin tissue of each group of subjects according to the present invention.

Fig. 2 is an image diagram of the immunostaining of the skin tissue of each group of subjects according to the present invention.

In order to facilitate the description of the central idea of the present invention expressed in the column of the above-mentioned summary of the invention, specific embodiments are used to express it.

The present invention provides a composition with antioxidant activity. The composition includes an effective dose of Rhodiola rosea extract and monoglycerophosphocholine and its pharmaceutically acceptable carrier or pharmaceutically acceptable salt.

Rhodiola is a plant belonging to many years of vegetation or shrubs. Rhodiola extract contains a lot of antibacterial and anti-inflammatory ingredients such as scoparidine and kaempferol, and it also has the effects of anti-anoxia, anti-fatigue, delaying aging and regulating the endocrine system.

Glycerophosphocholine (alpha-Glycerophosphocholine, alpha-GPC) is a small molecule nutrient. Glycerophosphocholine is mainly found in human cells and breast milk, or it can be extracted from non-based modified soybeans through deacetylation technology and special ion exchange technology. Studies have confirmed that glycerophosphocholine can help tissues grow and rebuild, activate organs, restore brain functions that lack circulation, and maintain physiological functions such as brain attention, concentration, and memory. According to current clinical trial studies, the oral route of glycerophosphocholine can increase the concentration of choline and acetylcholine in the blood, thereby reducing the concentration of cortison and further increasing the concentration of human growth hormone (hGH) in the body. the goal of.

Among them, the weight ratio of rhodiola extract: glycerophosphocholine is 1:1 to 1:6. In a preferred embodiment of the present invention, the weight ratio of rhodiola extract: glycerophosphocholine is 1 : 3.

In addition, the composition further includes a resveratrol and a guarana. Resveratrol is widely present in mulberries, peanuts, grapes and other plants, especially in grapes. Studies have found that resveratrol has excellent antioxidant activity, which not only protects the human body from free radicals, but also acts as a plant When exposed to environmental pressure, fungal and bacterial infections, antitoxins are also produced to fight against external attacks.

Guarana is an excitatory stimulating substance that can increase mental and muscle strength resistance, endurance, and can reduce exercise and physical fatigue. The xanthines contained in caffeine and caffeine in guarana can provide the fastest and most stable thinking.

The composition of the present invention can be made into powder, granular or liquid form, and can be processed to make the composition of the present invention into a capsule form; in particular, the capsule form can be an animal capsule or a plant Sex capsules. Animal capsules can be made of gelatin, and vegetable capsules can be made of sodium carboxymethyl cellulose, which are provided for oral consumption by users.

The term "pharmaceutically acceptable salt" used in the present invention includes water-soluble and water-insoluble salts, which can retain the biological efficacy and properties of the composition. The term "pharmaceutically acceptable carrier" as used in the present invention means a substance, composition or vehicle that involves the transportation or transport of a pharmaceutical preparation from one organ or part of the body to another organ or part of the body. It covers flavoring agents, flavor enhancers, preservatives, antioxidants, chelating agents, penetrants, lubricants, tablet adjuvants, coloring agents, humectants, binding agents, and equivalent carriers.

The examples of the present invention are verified by animal experiments, which prove that the composition of the present invention has the effects of inhibiting fat formation and antioxidant activity.

Animal experiment

(1) Animal breeding

One of the subjects in the experiment came from the National Laboratory Animal Center of the National Institutes of Health. The subjects used 60 male C57BL/6 (B6 ) mice aged 6-8 weeks with an initial weight of about 20 grams. The candidates are a group. Each group of subjects was kept in an animal room at 22±2°C, and the light time and dark time were controlled by a timer. The light time and dark time were 12 hours each. Since the subjects usually eat during the dark hours, the best time to remove the feed is when it starts to turn bright, and the fasting time can be set to 12±2 hours.

(2) Feeding

The subjects were given high-fat feed orally, and D-galactose was given by injection. It is worth noting that the high-fat diet is designed to induce weight and obesity in subjects; excessive injection of D-galactose will produce a large amount of reactive oxygen species in the subject’s body, thereby destroying the equilibrium state of the concentration of reactive oxygen species in the body. Induce peroxidation in the subject.

(3) Dosing setting

Subjects aged 6-8 weeks were randomly divided into six groups according to their body weight, each with 10 animals. Each subject was given high-fat diet and injected D-galactose in the animal room. The composition of the present invention containing rhodiola extract and glycerophosphocholine was administered orally, and the experiment was carried out for 12 weeks. During the experiment, the body weight was measured and recorded every week. The term "administration" as used in the present invention refers to the subject directly administering the composition or the pharmaceutically acceptable salt of the composition, which can form an equivalent amount of the active composition in the subject's body.

The experimental groups can be divided into: (1) Normal group: freely ingested normal feed, and given sterile reverse osmosis water 10ml/kg. (2) Control group: Free intake of high-fat feed, 0.3-1.2g/kg bw of D-galactose injected subcutaneously on the back of the neck and 10ml/kg of sterile reverse osmosis water. (3) Experimental group 1: Free intake of high-fat feed, subcutaneous injection of 0.3-1.2g/kg bw of D-galactose on the back of the neck; 1 times the dose of Rhodiola rosea extract, including the content of Rhodiola rosea extract It is 88.4 mg/kg body weight/day. (4) Experimental group 2: Free intake of high-fat feed, subcutaneous injection of 0.3-1.2g/kg bw of D-galactose on the neck and back; low-dose of the composition of the present invention (Rhodiola rosea extract: glycerophosphocholine = 1:1), wherein the effective dose of the combination of Rhodiola rosea extract and glycerophosphocholine of the composition of the present invention is 187 mg/kg body weight/day (Rhodiola rosea extract: glycerophosphocholine=1:1) , According to the body surface area ratio, the oral dose of 0.91 g/day is relative to the human body. (5) Experimental group 3: Free intake of high-fat feed, subcutaneous injection of 0.3-1.2 g/kg bw of D-galactose on the back of the neck; medium dose of the composition of the present invention (Rhodiola extract: glycerophosphocholine = 1:3), in which the group of the present invention The effective dose of Rhodiola rosea extract and glycerophosphocholine combination is 384.4 mg/kg body weight/day (Rhodiola rosea extract: glycerophosphocholine=1:3), according to the body surface area ratio, relative to human oral administration 1.87 g/day dose. (6) Experimental group 4: Free intake of high-fat feed, subcutaneous injection of 0.3-1.2g/kg bw of D-galactose on the back of the neck; administration of a high dose of the composition of the present invention (Rhodiola rosea extract: glycerophosphocholine = 1:6), wherein the effective dose of the combination of Rhodiola rosea extract and glycerophosphocholine of the composition of the present invention is 680.4 mg/kg body weight/day (Rhodiola rosea extract: glycerophosphocholine=1:6) , According to the body surface area ratio, the oral dose of 3.31 g/day is relative to the human body.

Conversion of test doses between experimental animals and humans: The conversion of doses between humans and experimental animals in this assessment method is in principle based on the estimation of the maximum safe starting dose in initial clinical trials for the US Food and Drug Administration in 2005. therapeutics in adult healthy volunteers), and based on a 60 kg adult, using experimental When the substance is tested, the effective dose conversion is in principle based on the human body’s daily recommended intake per kilogram of body weight (/kg bw/day) based on the equivalent dose.

Since animals increase their food intake as their body weight increases, the above two conversion methods have little difference in the daily intake of test samples obtained by animals, so both can be used. The feed for the test group and the control group must contain approximately the same amount of calories, protein, fat, calcium and other minerals or vitamins.

Conversion of animal test results into recommended doses for humans: The most suitable effective dose is calculated from the statistical results of comprehensive animal tests. If the unit is% in the feed, the same% is the total dietary intake of humans in a day (500 g dry weight). The% of) is 1 times the dose. For example, when 1% of the test product group in the feed obtains the best comprehensive result, it is considered that humans also take 1% (5 grams) as the best 1 times the dose. If the experiment is to use per kilogram of body weight (/kg b.w.) as the unit, it will be multiplied by 60kg as the effective dose for adults.

(4) Sampling analysis

(A) Weight change: During the experiment, the body weight should be weighed at least once a week (it should be fixed in the animal room every day before the light is turned dark and the feed is given), and the weight change of each group of subjects should be observed. The weight change conversion method is: weight change (weight change) = end weight-starting weight.

Please refer to Table 1, which is the body weight change table of each group of subjects in this embodiment. The results in Table 1 show that as the number of experimental weeks increases, the average weight of all experimental subjects has increased steadily. Among them, the average weight of the control group has increased the most; however, for the experimental groups 2 to 4, the After the subjects were given the composition of the present invention, their average body weight was significantly lower than that of the control group. Among them, in experimental group 3, the average weight of the subjects was significantly lower than that of the control group. The effect of grams; it is obvious that the present invention can effectively reduce the weight of subjects by mixing Rhodiola rosea extract and glycerophosphocholine, and also gives Rhodiola rosea extract: glycerophosphocholine =1: The experimental group 3 of 3 has the best overall weight loss effect.

(B) Changes in organ and body fat weight: When the animal is sacrificed, carefully remove the epididymal fat, kidney fat, and intestinal fat from the peritoneal cavity for precise weighing, and calculate the body fat percentage. The formula for calculating body fat percentage is as follows: body fat percentage = [body fat (g)/weight (g)]×100%. In the actual calculation, body fat mass (g) = epididymal fat (g) + kidney peripheral fat (g) + mesenteric fat (g). The determination of body fat should try to treat all animals with the same method by the same person in order to reduce the error between individual animals.

Please refer to Table 2, which is a table of changes in organ and body fat weight of each group of subjects in this embodiment. The results in Table 2 show that after 12 weeks of the experiment, the body fat percentage and total fat of all experimental subjects have an upward trend, of which the increase in the control group is the most obvious ( P <0.05); however, compared to For experimental groups 2 to 4, after subjects were given the composition of the present invention, their body fat percentage and total fat were significantly lower than those of the control group ( P <0.05); that is, the composition of the present invention The substance can effectively inhibit obesity and fat accumulation.

(C) Changes in various biochemical values in blood: Use blood biochemistry to analyze various biochemical values that are not easy to form body fat function in the body. The biochemical analysis items are as follows:

(a). Blood lipids: After 12 hours of fasting, each group of subjects was anesthetized with isofurane, collected blood from the celiac artery, centrifuged to collect serum, and then analyzed and detected the serum with a serum biochemical analyzer The concentration of triglycerides, cholesterol, low-density lipoprotein, and high-density lipoprotein.

(b). Liver lipids: After blood collection from the celiac artery, the abdominal cavity was flushed with saline, and then the body lipids were further extracted by the chloroform-methanol extraction method, and the concentrations of liver triglycerides and cholesterol were measured.

(c). Blood glucose: After the celiac artery blood sampling is completed, the blood glucose concentration in the serum is measured by the enzyme method and the colorimetric principle.

(d). Renal function: After blood collection from the celiac artery, the concentration of creatinine in the serum is measured by the enzyme method and the colorimetric principle.

Please refer to Table 3, which is the blood biochemical value change table of each group of subjects in this embodiment. As shown in Table 3 above, after 12 weeks of the experiment, the blood glucose results showed that the blood glucose concentration of the control group was significantly increased compared to the normal group ( P <0.05); however, the blood glucose concentration of the experimental group 1 to 4 was compared with that of the control group. The group is slightly lowered. Cholesterol results showed that the concentration of cholesterol in the serum of the experimental group 1 to 4 was significantly lower than that of the control group ( P <0.05). Creatinine results showed that the serum creatinine concentration of experimental group 1~3 had a slightly lower trend than that of the control group. Among them, the creatinine concentration of experimental group 4 was significantly lower than that of the control group ( P <0.05). The low-density lipoprotein (LDL) results showed that the concentration of low-density lipoprotein in the experimental group 1 to 4 was significantly lower than that in the control group ( P <0.05). The results of high density lipoprotein (HDL) showed that the concentration of high density lipoprotein in the experimental group 1 to 4 was significantly higher than that in the control group ( P <0.05). Thus, the experimental results in Table 3 show that the composition of the present invention can effectively inhibit the formation of multiple biochemical activities of body fat, and effectively achieve the effect of reducing body fat rate.

(D) Changes in blood antioxidant indicators: using enzyme method and colorimetric principle to measure the concentration of superoxide dismutase (SOD), glutathione peroxidase (GSH Px) and G6PD (Glucose-6-phosphate dehydrogenase) in serum The active concentration.

Please refer to Table 4, which is the blood antioxidant index change table of each group of subjects in this embodiment. As shown in Table 4 above, after 12 weeks of the experiment, the results of superoxide dismutase (SOD), glutathione peroxidase (GSH Px) and G6PD show that the experimental group 2~4 are significantly more significant than the control group The increase in sex resistance ( P <0.05), among which, the rhodiola extract of experimental group 3: glycerophosphocholine=1:3, superoxide dismutase (SOD) and glutathione Oxidase (GSH Px) and G6PD have the highest measured concentrations. Obviously, the present invention can effectively enhance the anti-oxidant effect by administering rhodiola extract and glycerophosphocholine, thereby achieving the purpose of eliminating peroxidized molecules and free radicals in the body. Experiments have confirmed that the experimental group 3 The antioxidant effect is the best, which in turn helps delay the aging effect.

(E) Changes in blood growth hormone: human growth hormone (hGH) analysis: human growth hormone ELISA kit was used to analyze the concentration of hGH in serum.

Please refer to Table 5, which is the growth hormone change table of each group of subjects in this embodiment. As shown in Table 5 above, after 12 weeks of the experiment, the results of growth hormone in the blood showed that the concentration of growth hormone in the serum of the control group was significantly lower than that of the normal group ( P <0.05), but the experimental group 1~4 have a significant increase compared to the control group ( P <0.05). Furthermore, the subjects were induced by aging and obesity, through the administration of Rhodiola rosea extract and glycerophosphocholine, the effect of the growth hormone in the body was relatively improved, and the effect of helping the subjects to lose weight was achieved. .

(F) Observation of staining of tissue sections

(1). After each group of subjects is induced to age with D-galactose, the changes in hair color should be observed. If there is aging, the hair will turn white, so it needs to be photographed and recorded. Please refer to FIG. 1 in conjunction with the figure, which is an image of the hair follicle and subcutaneous fat staining of the skin tissue of each group of subjects in this embodiment. As shown in Figure 1, compared with the normal group, the hair follicle tissue of the control group was significantly loosened, and the thickness of the subcutaneous fat of the control group was also significantly higher than that of the normal group; however, the experimental group 1~4 after the experiment It was found that compared with the control group, the hair follicle tissues of the experimental groups 1 to 4 gradually recovered to a dense state, and the proportion of subcutaneous fat thickness also decreased significantly.

Please cooperate with the results shown in Figure 1 and Table 6 above. It is further found that the subjects in experimental groups 2 to 4 who were given Rhodiola extract and glycerophosphocholine were compared with those in experimental group 1 who were given a single Rhodiola For the subjects of the extract, the thickness of the subcutaneous fat measured by the tissue sections of the experimental group 2 to 4 has a more obvious effect of reducing the subcutaneous fat. Among them, the reduction effect of the subcutaneous fat thickness measured by the experimental group 3 is the best . It is stated that the subjects given the combination of rhodiola extract and glycerophosphocholine can not only effectively increase the hair follicle tissue of the skin and increase the hair content, but also further reduce the thickness of the subcutaneous fat in the body. Purpose.

(2). Collect the skins of subjects in each group and immerse them in 10% formalin solution for fixation, To conduct 4-HNE (4-hydroxynonenal) staining experiments; in particular, the subjects are injected with D-galactose to induce lipid peroxidation in the body, and the lipid peroxidation process will produce malonaldehyde (MDA). ) And 4-hydroxynonenal (HNE), which in turn leads to changes in the fluidity and permeability of the cell membrane, as well as the cell structure and function; therefore, the present invention can be used in 4-HNE staining experiments to observe each Group of subjects’ peroxidation status, where the darker the color of the deposition measured by 4-HNE staining, the higher the peroxide content in the subject; in other words, the more the color of the deposition measured by 4-HNE staining When it is shallow, it means that the peroxide content in the subject is lower. Please refer to FIG. 2 with the figure, which is an image of immunostaining of the skin tissue of each group of subjects in this embodiment.

As shown in Figure 2, compared with the normal group, the 4-HNE staining experiment of the control group has a significantly darker color of the deposited amount. Oxidation phenomenon; however, for the experimental groups 2~4 after the test, the deposition color of 4-HNE dyed compared with the control group, the deposition color has a significantly lighter trend, and the experimental group 3 It is measured that the color of the deposition amount of 4-HNE dyeing is lighter, and the dyeing area of the deposition amount is smaller, which indicates that the anti-peroxidation effect of experimental group 3 is the best; therefore, the rhodiola extract of the present invention The combination with glycerophosphocholine can further achieve the effect and purpose of enhancing the anti-peroxidation effect.

In summary, the present invention has been experimentally confirmed that the combination of rhodiola extract and glycerophosphocholine can not only effectively reduce the weight and body fat of the subject, but also enhance the anti-peroxidation effect in the body. Among them, when the combination ratio of rhodiola extract and glycerophosphocholine is 1:3, the effects of inhibiting body fat, reducing body weight and anti-oxidation are the best; thereby, the composition of the present invention can As a medicine for treating obesity, it can also enhance the anti-peroxidation effect of tissues in the body and achieve the effect of delaying aging.

Therefore, the above-mentioned embodiments are only used to illustrate the present invention, not to limit the present invention. Of. All modifications or changes made without violating the spirit of the present invention fall within the scope of the present invention's intended protection.

Claims (9)

A composition with antioxidant activity, the composition comprising an effective dose of an active ingredient and a pharmaceutically acceptable carrier or pharmaceutically acceptable salt thereof, wherein the active ingredient is composed of a rhodiola extract and a monoglycerophosphate bile The weight ratio of the rhodiola extract: the glycerophosphocholine is 1:3. The composition with antioxidant activity according to claim 1, wherein the effective dose of the composition is 384 mg/kg body weight/day. The composition having antioxidant activity according to claim 1, wherein the composition is in the form of a capsule. The composition with antioxidant activity according to claim 1, wherein the composition is in the form of powder, granules or liquid. A use of a composition with antioxidant activity, which is used to prepare a product that enhances the antioxidant activity in the body, wherein the composition contains an effective dose of Rhodiola rosea extract and monoglycerophosphocholine and its pharmaceutically acceptable A carrier or a pharmaceutically acceptable salt, the weight ratio of the rhodiola extract: the glycerophosphocholine is 1:1 to 1:6. The use according to claim 5, wherein the indicator of the antioxidant activity is the active concentration of an enzyme in the serum, and the enzyme is superoxide dismutase (SOD), glutathione peroxidase (GSH Px), or Its combination. The use according to claim 5, wherein the weight ratio of the rhodiola extract: the glycerophosphocholine is 1:3, and the indicator of the antioxidant activity is the active concentration of the enzyme in the serum, and the enzyme is G6PD (Glucose-6-phosphate dehydrogenase). The use according to claim 5 or 6, wherein the rhodiola extract: the glycerophosphoric acid When the weight ratio between choline is 1:1 to 1:6, the effective dose of the composition is 187 to 680 mg/kg body weight/day. The use according to any one of claims 5 to 7, wherein when the weight ratio of the rhodiola extract: the glycerophosphocholine is 1:3, the effective dose of the composition is 384 mg/kg Weight/day.

Images