TW201100088A - Extraction methods and compositions to improve hyperlipidemia, hyperglycermia and fatty liver - Google Patents

Abstract

By using PPAR α and PPAR γ transactivation assay, we successfully identified the extraction methods and composition of bitter gourd, licorice, soybean protein, and chlorella could exert the highest activation ability of PPAR α and PPAR γ. We further demonstrated the efficacy in animal models and human models to ameliorate hyperlipidemia, hyperglycermia and fatty liver.

Description

201100088 VI. Description of the invention: [Technical field of invention] A formula combination of a health food and a method for extracting each material [Prior Art] In a developed country, 'lipid and lysate metabolism is poor, and symptoms of metabolic syndrome are easily generated. Such as diabetes, hypertension, hyperlipidemia, arteriosclerosis, high triglycerides and other diseases, but recent studies have found that 'these diseases, contact with the sputum receptor - ppARs (liposome activated proliferative receptors). Many scientists are currently looking for activators that can activate ppARs as drugs for metabolic syndrome, such as: thiazolidiiiediones (TZD) (Tengdao sensitizer) has the effect of reducing ash sugar by activating PPARr, while fibrates The two types are activated PPARa and have the effect of lowering blood fat, but since all of the above are drugs, they have side effects and are not suitable for long-term consumption. 〃 In recent years, there has been a small amount of literature on the hypoglycemic materials commonly used in ancient times to verify whether the effect of lowering blood glucose is achieved by activating PPARr, and the natural ingredients can be found to activate PPAR compounds for the purpose of Drug development. At present, the materials in the literature are mainly unilateral. For example, hop extracts can simultaneously activate ppARα and ^, bitter gourd extract can simultaneously activate retort and r, licorice extract activation ρρ break, etc., but these active ingredients often need to go through Special organic solvents are extracted to obtain effective scores. However, in the concept of food, it is not suitable for the food to be tested after organic dissolution. Therefore, this patent is to use the special alcohol and / or water extraction and the best ratio of each group of materials to determine the most effective (lipid, blood sugar, fatty liver) food compound formula. 201100088 【Abstract】 At present, many drugs for lowering blood sugar and lowering blood fat are developed through activation of PPARa and r. Therefore, the principle of efficacy and mechanism has been confirmed by many experts and scholars, but this kind of medicine is mainly used in gj. Synthetic compounds, which cause some subjects to produce liver or kidney toxicity', thus making the body's GOT (gumaceous acid acetic acid transaminase), GPT (Chu amino acid pyropolysaccharide) higher, so The idea of this patent is to use the medical community to find a way to activate PPAR activators. For the materials that humans often eat, the foods can be screened for foods to find the activated pp plate and r natural compounds or compound combinations for the benefit of more. People. 〇本创意 疋 A multi-effect combination of foods with blood sugar lowering and improved fatty liver and its pure method, which are added and subtracted as bitter melon 20~40%, licorice 1%~10%, green algae 15%~ 30%, soy protein powder 2 (M〇%. The extraction method of each material is as follows: the remaining HL bitter melon first _ juice machine _ take bitter melon juice, left ΐ ί ί ί ί 200 200 200 200 200 200 200 200 200 After 8 hours, ssm 混合 mix the bitter gourd juice and alcohol extract for cold drying ' 乙 Ο 丙 丙: 100 grams of licorice after crushing, add 500~1 〇 _ volume Ϊ pure ϋ 100, shake at room temperature 10 After more than an hour, remove the licorice residue and read the cold-dried powder into a powder. This powder is then dried into a powder by soaking the alcohol-soluble portion with 5 (Η_methanol soaked for 20i, i or more). , pumping 500~1000ml of 100°c water for the pumping time to filter the aqueous solution with l分子mm-5 coffee f·molecular sieve, removing the residual > an aqueous solution portion for concentration and drying and dusting. ', Π : 100 grams of degreased soybean meal, with 300~5_ of water, add ΐ^ ff 丨 〇 ^〇_idase (EC 3.4. 24. 28), at 10000 DaTtiTT^ 5 The above formula was used for animal and human trials, which confirmed the effects of lowering blood fat, blood sugar and improving fatty liver, and the combination of the above processes has good effect and good water diffusion. The 'ability' can be considered for other compound food combinations, including the development of capsule lozenges, Gu 201100088 powder, beverages, oral liquid products. [Embodiment] The method of transfecting PPAR 0: and PPAR r cells (transactivation assay system) was established. (transfected cell activation system) - accessing the luminescent signal behind the activated PPAR promoter gene, then sending the gene to the expressed liver cells, and then using the material to stimulate the cells, when the material has the ability to activate PPAR, That is, the intensity of the cold light can be measured by the luminometer to determine the effect of the material activation PPAR α and PPAR «r, and the technique can also be used to determine the process and the combination of the combinations.

After screening many food materials, the following four materials were found to have good activity 'and different extraction methods' to find the following patented processes, which have the best activation ppAR effect, and the manufacturing methods are as follows: A, bitter gourd extract :1 新鲜 新鲜 fresh bitter gourd first extract the bitter gourd juice with juicer, and the remaining slag is soaked in 100~200 ml 90% alcohol for 6-8 hours, filter to remove the residue, extract bitter gourd juice and alcohol The mixture was lyophilized and made into a powder. Ethyl propylene. Licorice extract: 1 gram of licorice after pulverization, add 5 〇〇 ~ i 〇〇〇 mi volume of pure water to 6 (M 〇〇rpin shaking at room temperature for more than 1 hr, after removing licorice residue The upper π liquid is cooled; the east is dried into a powder, and the powder is further immersed in 50-100 ml of methanol for more than 20 hours, and the alcohol-soluble portion is further dried into a powder. ' Chlorella: 100 g of broken green algae powder, 50 (M000 mllOOt: boiling water for 60 to 180 minutes, the aqueous solution is filtered by molecular sieves to remove the residue, and the aqueous solution is partially concentrated and dried to be dusted. " Ding, soy protein powder: 100克油油豆粕, with 300~500ml of water, adding 0.3%~3% proteolytic enzyme end〇peptidase (EC 3. 4. 24· 28 ), acting at 45-50 degrees for 6-8 hours, After filtration, the filtrate is taken, and the components below 10000 Dalton are filtered by molecular sieve. 1 Then, the combination and combination of the compounds are used, and the materials extracted by the above four patent processes are used to find the best dose-inducing activity, and the material is also used. Arrange and combine, the experimental results found that these four materials When combined, the PPARa and ppAR τ cell line screening methods have better ability to activate PPARa and PPARγ than any combination of unilateral or other formulations (compared to the same total concentration) (see Figure 1 and Figure 2 below). Figure 1: Represents the multiple of activated PPARa, Figure 2 shows the multiple of activated PPARr, while the control group is 201100088. The result of using only water to stimulate cells is '100% (1 fold), while licorice, bitter melon, green algae The maximum activation degree of ΡΡΑΚα is only 1.5, L4, 2 3, 2.1 times. When the four components are combined, the maximum activation degree of ppARa can be as high as 5.3 times' and the maximum activation degree of four unilateral pairs of ppARr respectively 2.9, 1.3, 1.8, 1.9 times' When the four components are combined, the maximum activation degree of ppARr^ is up to 5 times. The above results confirm that the compound combination has the effect of adding ppARa and hydrazine, which is superior to unilateral. The combination has its progressiveness. I (multiple water (control group > licorice *:- fine soy called! jie side 3! Activate PPARa*, Uv 1.4^ i 2.3^ 2丄, 5.3^ 丨 piSi: T^ 1^ . ......-................ u,iik 1,9^ 5.0^ 〇 〇

Control ^ 100% 200% 300% <100% 500% m / riu

184 184% IHiim

Control 0% 100% 2003⁄4 300% 400% 5003⁄4 600?ό FUf / RUi Figure 1 The above formula is further tested in vivo for blood lipids, blood sugar and fatty liver to further confirm its efficacy 201100088 _ blood lipid blood glucose animal experiment: The C57BL/6 strain mice purchased by the Chengdu Animal Center were induced by high-fat diet to cause hyperglycemia and hyperlipidemia. After we designed the mice, we found that the mice were fed after 3 weeks. It has the effect of lowering blood sugar (14.6 %) and triglyceride (21.8%), while in the sixth week, the effect of lowering cholesterol (12.4%) can be seen. _ Blood lipids and blood sugar human experience: In the blood sugar experience part, after one month of taking, the average blood glucose level decreased from 99.1 mg / dl to 94.3 mg / dl (decreased 5%). In the blood lipid experience part, after one month of testing, the average cholesterol value of all experiencers decreased from 233 mg/dl to 207 mg/dl (a decrease of 11.1%); while the average value of triglycerides was 221 mg/dl. Dropped to I62mg/dl (a decrease of 26.7%), the experience has a 100% satisfaction. During the experience, the experiencer's liver index (breast amino acid acetic acid transaminase and amino acid pyrogal aminotransferase) and kidney index (creatinine) did not rise or even decreased, confirming that the patented formula would not cause liver and kidney. The burden. When the experiencer does not continue to take the patented formula (more than one month), it can be seen that the blood sugar and blood lipids (except triglycerides) of the experience have rebounded, confirming that the blood sugar and blood esters are the result of the patented formula. Patent compound vs blood sugar change = 23)

7 201100088 Patent compound VS triglyceride change (N=21) =3 "bJl s 400 350 300 250 200 150 100 50 0 〇〇1 η Ί 1^1 Q 17J 0_ 1 2 month patent before use Compound VS cholesterol change (N=23)

1 month 2 months before use Liver index 80 60 40 20 0 300

GOT '1 3:.8 3;.2

1 month before consumption □ 2 months

GPT

8 201100088 Fatty Liver Animal Experiment··C57BL/6 strain mice purchased by Chengda Animal Center, induced by high fat and high fructose feed, and added alcohol to drinking water to make the symptoms of fatty liver in mice. Formulated to feed the mice, you can find that after 2 months of feeding, the rats can reduce the liver index (brome amino acid Jiao grape to wind-based enzyme), and the patented formula of the bran amino acid Jiao grape transaminase index from 50 U / L At 36 U/L, the transaminase index of the amino-based sorghum grape without the patented formula control group increased from 51 υ/L to 55 U/L. In addition, it also sacrificed the old take, which can be found in the liver of mice. The average level of liver triglyceride in the formula rats is 23 mg, and the control group without the formula is 28 mg on average, and there are significant differences, while the mouse abdomen and subcutaneous The traceability of the formula is significantly lower than that of the non-drinking formula, so this patented formula has the effect of reducing fat accumulation.

201100088 Fat distribution ratio 30 r

Control □ patent compound total fat month complex fat subcutaneous fat fl fat liver human body difficult: this special gamma for the human body, three off can see the average biochemical value of the effect of the flap, including the money grade grass acetic acid transaminase ( Reduced from 46 U/L to 30 U/L), bran amino acid pyropolyaminogen (from muscle to 35 muscle), triglyceride (from 2〇4 mg/dl down to i69 tons he ), total cholesterol (from 220mg / dl t weeks to 2 〇〇 mg / dl) have significant improvement, even the experience of the New Zealand can be secret 3 ribs, φ super reading _ towel fat or light Degree of fatty liver. Take two

GPT anomaly (N=12)

After the detailed deletion of 201100088, this patent formula is different from the product that only acts on the gastrointestinal tract 'inhibiting oil absorption' on the market, and there is no product on the market that simultaneously appeals to lowering blood stasis and lowering blood fat and improving the efficacy of fatty liver. The formula and the extraction method of each material are patents with novelty, progress and industrial utilization. [Simple diagram description] [Main component symbol description]

Claims (1)

201100088 VII. Scope of application for patents: 1. A multi-effect combination of foods with blood sugar lowering and fatty liver, which is characterized by a combination of bitter gourd, green algae, licorice and soy protein powder. 2. A method for extracting each material extract of a multi-effect compound food combination having blood sugar lowering blood fat and fatty liver. 3. According to the multi-effect compound food composition described in Item 1 of the patent application, the addition ratio is: 20~40% of bitter gourd, 1%~10% of licorice, 15%~3〇% of green algae, soy protein powder 2 (μ4〇0/〇. 4. The method for extracting the materials according to item 2 of the patent application scope is as follows: A, Gourd extract: 100 g of fresh bitter gourd, first extract the bitter gourd juice with a juicer, and After the slag is immersed in 100~200 ml of 90% alcohol for 6 to 8 hours, the residue is removed, and the bitter gourd juice and the alcohol extract are mixed and freeze-dried to produce ethyl acetoacetate: 100 g of licorice is crushed. Add 500~1000ml volume=pure to 6(H〇〇rpm, shake at room temperature for more than 1 hour, remove the licorice residue, freeze the liquid into a powder, and then soak the powder for 50 hours with 50~100ral sterol. The alcohol-soluble fraction is further dried into a powder. The U00 gram broken green algae powder is discharged at 500-1000 ml l〇〇t: boiling water for a minute. The aqueous solution is filtered at a molecular sieve of 1 Gmm-5 mm to remove hydrazine. The wound 'aqueous solution part is concentrated and dried to spray powder. η visual powder: 100 grams of oil removal Bean cypress '300~500ml of water, added decomposing enzyme endopeptidase (EC 3· 4. 24. 28), Yu loo loooo ? J Hi specializes in 1 Xuxiang Xiangfang food 1^ composition, its product type There are four types of capsules, capsules, powders and oral liquids. 12 201100088 IV. Designation of representative drawings: (1) The representative representative of the case is: (No). (2) Symbols of the representative figure Brief description: None 5. If there is a chemical formula in this case, please disclose the chemical formula that best shows the characteristics of the invention: