阅读说明：本技术 橘皮素在制备控制人体血尿酸水平的药物或食品中的新用途 (New application of hesperetin in preparation of medicine or food for controlling blood uric acid level of human body ) 是由 周志钦 李真晴 刘猛 于 2021-02-05 设计创作，主要内容包括：本发明属于药品、医用食品、保健品、功能食品或食品领域,涉及橘皮素的新用途,具体涉及橘皮素在制备用于控制人体血尿酸水平的药物、医用食品、保健品、功能食品或食品中的用途；尤其是橘皮素在制备用于调节运动状态下血尿酸升高的药物、医用食品、保健品、功能食品或食品中的用途。(The invention belongs to the field of medicines, medical foods, health products, functional foods or foods, and relates to a new application of hesperetin, in particular to an application of hesperetin in preparing medicines, medical foods, health products, functional foods or foods for controlling the level of human hematuria acid; in particular to the application of the hesperetin in preparing medicines, medical foods, health products, functional foods or foods for regulating the rise of blood uric acid in a sports state.)

1. Use of hesperetin in the preparation of drugs, medical foods, health foods, functional foods or foods for controlling the level of human hematuria.

2. Use according to claim 1, characterized in that: the control of the blood uric acid level of the human body is to maintain the blood uric acid stability;

further, the control of the blood uric acid level of the human body is to control the rise of blood uric acid in a motion state;

further, the control of the blood uric acid level of the human body is to regulate the rise of blood uric acid caused by stress reaction in a motion state;

further, the control of the blood uric acid level of the human body is to reduce the synthesis of uric acid in a motion state;

further, the control of the blood uric acid level of the human body is to promote the excretion of uric acid.

3. Use according to claim 1, characterized in that: the medicine is an oral preparation or an injection preparation which is prepared by taking the hesperetin as an active ingredient and adding pharmaceutically conventional or acceptable auxiliary materials;

further preferably, the oral preparation is tablet, powder, granule, tea, capsule, soft capsule, oral liquid, pill, paste, or medicated wine;

further preferably, the injection preparation is an injection.

4. Use according to claim 1, characterized in that: the health product is an oral preparation prepared by taking the hesperetin as an active ingredient and adding conventional or acceptable auxiliary materials on the health product;

further preferably, the oral preparation is tablets, powder, granules, tea, hard capsules, soft capsules, oral liquid, pills, paste, beverages, vinum, biscuits, candies, cakes, liquid milk and compound nutrients.

5. Use according to claim 1, characterized in that: the functional food is prepared by taking the hesperetin as an active ingredient and adding conventional or acceptable auxiliary materials on the food;

further preferably, the functional food is soft capsules, hard capsules, oral liquids, tablets, powders, pills, pastes or beverages.

6. Use according to claim 1, characterized in that: the medical food is prepared by taking hesperetin as an active ingredient and adding conventional or acceptable auxiliary materials on the food;

further preferably, the medical food is a medical nutrition, a dietary supplement for human use and an infant food;

further preferably, the medical food is soft capsules, hard capsules, oral liquid, tablets, powder, pills, paste and beverages.

7. Use according to claim 1, characterized in that: the food is prepared by taking the hesperetin as a raw material and adding auxiliary materials or additives which are conventional or acceptable in food science.

8. An hesperetin oral preparation is characterized in that: the oral preparation is prepared by taking the hesperetin as an active ingredient and adding conventional or acceptable auxiliary materials in pharmacy, health-care food science, functional food science and medical food science.

9. An hesperetin injection preparation is characterized in that: the hesperetin is used as an active ingredient, and pharmaceutically conventional or acceptable auxiliary materials are added to prepare the injection preparation.

10. An orange peel food, which is characterized in that: the food is prepared by taking the hesperetin as a raw material and adding auxiliary materials or food additives which are conventional or acceptable in food science.

Technical Field

The invention belongs to the field of medicines, medical foods, health products, functional foods or foods, and relates to a new application of hesperetin, in particular to an application of hesperetin in preparing medicines, medical foods, health products, functional foods or foods for controlling the level of human hematuria acid; in particular to the application of the hesperetin in preparing medicines, medical foods, health products, functional foods or foods for regulating the rise of blood uric acid in a sports state.

Background

Citrus is the first fruit in China and the world, and tangeretin (tangeretin) is one of the polymethoxylated flavone monomeric compounds rich in the peel of citrus fruits. According to the existing literature reports, the hesperetin has various important biological activities, such as anti-inflammatory, antioxidant, anticancer, cardiovascular and neurodegenerative disease prevention and the like. Compared with other flavonoid monomers, the hesperetin has high oral utilization rate (21.38%), good drug property (0.43: the possibility that the hesperetin becomes a drug is 43%), is green and natural, has extremely low toxic and side effects, and is very suitable for human bodies. Research teams of the applicant have long conducted research on citrus flavonoids, particularly polymethoxylated flavonoids, and human health, and have conducted exploratory research on the application of citrus active ingredients such as hesperetin to athlete health. Cell and animal experiments of a team of the applicant find that the hesperetin can improve exercise tolerance of a mouse, enhance antioxidant enzyme activity and improve the antioxidant capacity of an organism so as to reduce oxidative stress injury induced by exhaustive exercise. Meanwhile, research teams of the applicant also find that after 4 weeks of hesperetin (200mg/d) is supplemented to a human body, subjective fatigue of weight lifting athletes and sprinters is obviously eliminated, and serum cortisol content and uric acid level are obviously reduced. These results indicate that hesperetin may have a positive effect on improving physical performance and lowering uric acid levels in athletes. Therefore, the following inventions have been made on the basis of the above findings.

Disclosure of Invention

The invention solves the technical problem of providing a new application of hesperetin. In particular to the application of the hesperetin in preparing medicines, medical foods, health products, functional foods or foods for controlling the level of human hematuria.

Further, the invention provides application of the hesperetin in preparing medicines, medical foods, health-care products, functional foods or foods for controlling the blood uric acid level of a human body.

Further, the control of the blood uric acid level of the human body is to maintain blood uric acid stability.

Further, the control of the blood uric acid level of the human body is to control the rise of blood uric acid in a motion state.

Further, the control of the blood uric acid level of the human body is to regulate the increase of blood uric acid caused by stress response in the exercise state.

Further, the control of the blood uric acid level of the human body is to reduce the synthesis of uric acid in a motion state.

Further, the control of the blood uric acid level of the human body is to promote the excretion of uric acid.

The application is realized, and particularly, the synthesis of uric acid in a motion state is reduced and the excretion of uric acid is promoted by the hesperetin, so that the aim of controlling the rise of blood uric acid is finally realized; further improving the recovery ability of the organism after exercise, reducing the stress reaction after exercise and being beneficial to the recovery of the body function.

In order to better utilize the new application, the inventor of the invention prepares the hesperetin into medicines, medical foods, health-care foods, functional foods or foods so as to be pushed to specific production application.

The prepared medicine is an oral preparation or an injection preparation which is prepared by taking the hesperetin as an active ingredient and adding pharmaceutically conventional or acceptable auxiliary materials; the oral preparation is tablet, powder, granule, tea, capsule, soft capsule, oral liquid, pill, paste, medicated wine, etc.; further preferably, the injection preparation is an injection.

The prepared health care product is an oral preparation which is prepared by taking the hesperetin as an active ingredient and adding conventional or acceptable auxiliary materials on the health care product. The oral preparation is tablet, powder, granule, tea, hard capsule, soft capsule, oral liquid, pill, paste, beverage, medicated wine, cookies, candy, cake product, liquid milk, and compound nutrient.

The prepared functional food is prepared by taking hesperetin as an active ingredient and adding conventional or acceptable auxiliary materials or additives on the food; the functional food is soft capsule, hard capsule, oral liquid, tablet, powder, pill, paste, or beverage.

The medical food is prepared by taking the hesperetin as an active ingredient and adding conventional or acceptable auxiliary materials on the food; the medical food is medical nutriment, human dietary supplement and infant food; in particular, the composition can be soft capsules, hard capsules, oral liquid, tablets, powder, pills, paste and beverages.

The prepared food is prepared by taking the hesperetin as a raw material and adding auxiliary materials or additives which are conventional or acceptable in food science.

In the above-mentioned application, the specific product form is as follows:

an hesperetin oral preparation is characterized in that: the oral preparation is prepared by taking the hesperetin as an active ingredient and adding conventional or acceptable auxiliary materials in pharmacy, health-care food science, functional food science and medical food science.

An hesperetin injection preparation is characterized in that: the hesperetin is used as an active ingredient, and pharmaceutically conventional or acceptable auxiliary materials are added to prepare the injection preparation.

An orange peel food, which is characterized in that: the food is prepared by taking the hesperetin as a raw material and adding auxiliary materials which are conventional or acceptable in the food science.

The research of the invention shows that the product prepared by taking the hesperetin as the main raw material or the active ingredient before the exercise has the obvious effect of maintaining the stability of blood uric acid, particularly has the effect of controlling the rise of the blood uric acid in the exercise state, reduces the generation of the uric acid so as to reduce the stress reaction of the body caused by high-strength exercise, simultaneously promotes the excretion of the uric acid, reduces the internal environment of the body after the exercise, improves the content of urea, creatinine and creatinase, improves the oxidation resistance of the body, increases the muscle content of the body, improves the fatigue state after the exercise, is particularly suitable for athletes, and provides a brand-new sports supplement with remarkable safety effect for the public.

Drawings

Figure 1 subject cohort (a) and experimental schedule (B).

FIG. 2 serum uric acid levels of all subjects in the experimental and control groups (FIGS. 2A-2D).

Wherein: FIG. 2A experimental group female subjects serum uric acid levels; FIG. 2B serum uric acid levels in experimental male subjects; FIG. 2C serum uric acid levels in control female subjects; FIG. 2D serum uric acid levels in male subjects of the control group.

FIG. 3 Urea levels in all subjects in the experimental and control groups (FIGS. 3A-3D).

Wherein, FIG. 3A experimental group female subjects serum urea levels; FIG. 3B serum urea levels in male subjects in the experimental group; FIG. 3C serum urea levels in control female subjects; FIG. 3D control group male subjects serum urea levels.

Figure 4 creatinine levels in all subjects in the experimental and control groups (figures 4A-4D).

Wherein, figure 4A experimental group female subjects serum creatinine levels; FIG. 4B serum creatinine levels in experimental group male subjects; FIG. 4C serum creatinine levels in control female subjects; FIG. 4D control group male subjects serum creatinine levels.

Figure 5 creatine kinase levels in all subjects in experimental and control groups (figures 5A-5D).

Wherein, figure 5A experimental group of female subjects had serum creatine kinase levels; FIG. 5B serum creatine kinase levels in experimental group male subjects; FIG. 5C serum creatine kinase levels in control female subjects; FIG. 5D serum creatine kinase levels in control male subjects.

Detailed Description

The following is a verification test of the novel use of hesperetin according to the invention.

1. Research method

1.1 test subjects

22 sprinters (11 each of men and women) of a Chongqing city professional team are selected as test subjects, and the motion grades are all at the second level and above of the state; furthermore, all athletes were selected from the same large training set and used similar training protocols during the experiment. The study adopted a pairing, random experimental design. First, the subjects were randomly divided into an experimental group (height 171.98. + -. 5.14cm, weight 62.22. + -. 12.82kg, age 24.30. + -. 3.39years) and a control group (height 174.84. + -. 8.54cm, weight 57.92. + -. 9.81kg, age 23.10. + -. 2.76years) according to sex, shape and exercise grade (FIG. 1A). The control group received placebo: american Cansure whey protein isolate powder (purity 95%, stimulant detection number: 2019FD279), 20 g/d; the experiment group took the hesperetin supplement (20 g/bag, containing 19.8g of whey protein isolate with the purity of 95% and 200mg of hesperetin with the purity of 99.79%; production permit number: SC 10611310113704; stimulant test number: 2019FD234), 20 g/d.

Before participating in the study, all subjects were subjected to systematic medical examination, and patients with depression, cardiovascular disease, cushing's syndrome and serious disease in the last five years were excluded. Subjects were informed of the content, flow and potential risks of the study and signed informed consent. The experimental methods adopted by the institute strictly comply with the declaration of Helsinki and are approved by the academic and ethical committees of Shanghai sports academy.

1.2 data acquisition and processing

The study was conducted in a three-month winter training session of 2019 with 4 weeks of selection. All subjects were asked to enter the laboratory at 7:30-8:30 am daily and to receive the dispensed hesperetin supplement or placebo on site, as numbered, all under the supervision of the investigator. Blood samples of the subjects were collected on the first day of each week (T1, T2, T3, T4) and the next day after the end of the experiment (T5) (FIG. 1B), 4ml of venous blood was collected each time, each subject was in a fasting state (fasting was not less than 8 hours) before blood collection, and supplements or placebo was taken immediately after the end of blood collection. Within 30min after blood sample collection, serum samples were separated using a high speed centrifuge (Sichuan TG16, China) (2000R/min,15min), and then stored in a-80 ℃ medical refrigerator (Boke BDF-86V158, China). After the intervention experiment, a researcher performed a unified analysis of serum samples (mai rui SAL-6000, china; BioTek-Epoch, usa) and recorded serum uric acid, urea, creatinine and creatine kinase results. In addition, body composition tests were performed on all subjects at the beginning (T1) and end (T5) of the experiment to obtain data on body weight, body fat rate, fat mass, muscle mass, and protein ratio. During the test, the subject was asked to stand still on a body composition tester (InBody 570, korea) with light clothing.

Throughout the experiment, the diet of each subject was strictly controlled and recorded in detail. Meanwhile, a restaurant in a sports training center in Chongqing City is responsible for processing and distributing food materials. One researcher gives guidance and supervision on selection, cooking and the like of food materials, and removes food materials and seasonings which may affect the research result. In addition, to minimize interference from other factors, all subjects were prohibited from taking any other supplements and medications, etc., other than the indicated sports drink (best, usa), from 2 weeks prior to the start of the study to the end of the study (total 42 days). During the course of the study, all subjects did not show any symptoms of discomfort.

1.3 statistical analysis

Statistical analysis was performed by SPSS 25.0 software and data are presented as mean ± standard deviation. The subjects were analyzed for various body composition indicators (body weight, body fat rate, fat mass, muscle mass and protein ratio) and biochemical indicators of body function (serum uric acid, urea, creatinine and creatine kinase) using two-factor (2 groups x different time points) repeated measures of variance. If significant difference exists, pairwise comparison is carried out on the basis of LSD standard. Significance level was determined as p < 0.05.

2 results of the study

2.1 Effect of supplementation with 4 weeks of hesperetin on body composition

The body composition (body weight, body fat rate, fat mass, muscle mass and protein ratio) before and after 4 weeks intervention test in the test group and the control group are shown in table 1. Before the experiment begins, the experimental group and the control group have no statistical difference; at the end of the experiment, the body weight and muscle mass of the experimental group were significantly higher than those of the control group, but their body fat rate was significantly lower than that of the control group. The weight of the experimental group is increased by 0.42Kg, but the muscle is increased by 0.85Kg, and the fat amount is reduced by 0.4Kg, while the weight of the control group is increased by 0.14Kg, the muscle is increased by 0.13Kg, and the fat amount is not obviously changed. During winter training, the increase of the body weight of the control group is only the muscle growth caused by training, and under the intervention of hesperetin, the fat content of athletes is greatly reduced, and the muscle synthesis is promoted. These results indicate that, under certain conditions, hesperetin promotes fat metabolism and muscle formation, improves body composition of athletes, and enhances exercise performance.

2.2 Effect of 4 weeks of supplementation with hesperetin on Biochemical indicators of body function

The changes of various biochemical indexes (serum uric acid, urea, creatinine and creatine kinase) in the intervening experiment process of 4 weeks in the experimental group and the control group can be shown in table 2. Before the experiment (T1), the biochemical indexes of the experimental group and the control group have no significant difference. During the whole intervention test period, the serum uric acid level of the experimental group is in a whole descending trend, and particularly, the change of the male athletes (figure 2B) is obvious; the uric acid level of the control athletes is slightly increased, and the fluctuation is large.

From the examination of creatinine, urea and creatine kinase in all subjects in the experimental group (fig. 3-5), it can be seen from fig. 5 that the overall decrease in urea levels in subjects following hesperetin intervention is more pronounced, especially in male athletes (fig. 5B), which may be the result of a greater metabolic abundance in men than in women. In the female athlete group (fig. 3A), urea fluctuated, especially in the period from T3 to T4, and began to fluctuate, but remained stable as a whole.

In addition, the creatinine levels of all the subjects in the experimental group (fig. 4) were overall stable and slightly decreased, and the fluctuation still appeared in the period from T3 to T4, which is probably the internal environment fluctuation caused by longer holiday time after the training of the period T2. Similarly, the serum creatine kinase of the subjects in all experimental groups steadily decreases (fig. 5), which shows that the hesperetin intervention can effectively reduce the creatine kinase content and the occurrence of muscle damage, and particularly shows that the significant decrease of T1-T3 is shown in the subject #3 (fig. 5B), and the creatine kinase content of the subject #10 is greatly increased and then rapidly recovered to the normal level in the period from T2 to T4 (fig. 5A), and further shows that the hesperetin intervention effect is significant, the reaction is rapid, and the protective effect on body muscles is good.

Table 1 effect of supplementation with hesperetin on body composition

Note: compared to the initial value (T1), #, p < 0.05; p <0.01 compared to control group.

TABLE 2 Effect of Replementing hesperetin on serum uric acid, cortisol, creatinine and creatine kinase in the body

3. Discussion:

the study was aimed at discussing the effects of hesperetin supplementation on serum uric acid, urea and other relevant biochemical indicators (creatinine and creatine kinase) and body composition during winter training of sprinters. Researches show that after 4 weeks of supplementation with hesperetin (200mg/d), the serum uric acid level of athletes is remarkably reduced and gradually tends to be stable, and meanwhile, the serum urea content is remarkably reduced, and creatinine and creatine kinase change slightly and gradually tend to be stable; at the same time, body weight, muscle mass and protein ratio increased significantly, and body fat rate and fat mass decreased significantly. The results prove that the hesperetin can reduce the serum uric acid level and reduce the accumulation of uric acid in the body. This fully demonstrates that 4 weeks of hesperetin supplementation can reduce the stress response of the body caused by high intensity exercise, effectively reduce the level of hyperuricemia caused by stress of the body, improve body composition, and regulate body balance.

Uric acid is the end product of human food-borne and endogenous purine metabolism, and is produced primarily in the liver and small intestine by xanthine oxidase/xanthine dehydrogenase (XO/XDH). In most mammals, uric acid is further oxidized to allantoin by Uricase (UO), but human beings have evolved to inactivate uricase due to genetic mutations. Uric acid produced by the human body is excreted with urine and is subjected to complex treatment by the kidney, including glomerular filtration, secretion, reabsorption and the like. Uric acid has some antioxidant properties and can scavenge singlet oxygen, free radicals, peroxynitrite, and transition metal chelates, but uric acid can also be an oxidant when its chemical microenvironment changes. The antioxidation of uric acid is supported by many scholars, uric acid is also listed as a main non-enzyme antioxidant in some monographs, and uric acid is also used as a marker reflecting the antioxidation capability of organisms in some clinical researches. Research shows that in vitro environment, uric acid can directly eliminate active oxygen in vivo. Studies have also shown that oxidative stress caused by strenuous exercise can be alleviated by injecting uric acid into a human body before the exercise is done at high intensity. Therefore, uric acid acts as an antioxidant in the body under short/long exercise conditions, and its serum level gradually rises and even accumulates excessively high. Similar to the above research, the research finds that the uric acid level in vivo of athletes is gradually increased after winter training begins, and the uric acid level of athletes in an experimental group is obviously lower than that of an athletic group (fig. 2) after winter training is finished, which indicates that hesperetin has an obvious effect of reducing hyperuricemia caused by exercise, and the hesperetin has good application potential as an athletic supplement.

The uric acid synthesis pathway in the human body mainly comprises two major processes, namely synthesis and degradation of purine. Purine synthesis processes are further divided into de novo synthesis (de novo synthesis) and salvage synthesis (solvent pathway). Of which the de novo synthetic pathway dominates. Whereas the degradation process of purines starts mainly with the removal of amino groups. Adenosine deaminates to form inosine under the catalysis of Adenosine Deaminase (ADA). Guanine is deaminated to hypoxanthine catalyzed by Guanine Deaminase (GDA). Hypoxanthine and xanthine are subsequently oxidized under the action of XO to give uric acid. When people take a large amount of foods rich in adenine and hypoxanthine for a long time, the activities of deaminases such as ADA and GDA and XO are promoted, a large amount of uric acid is produced, and the serum concentration is increased. Similarly, under intense and violent exercise, the carbohydrate and fat in the body are oxidized by oxygen to generate a large amount of energy, so that the energy supply of the body is realized. In the process, a substrate for synthesizing the uric acid, namely Adenosine Triphosphate (ATP), is generated in a large quantity, and the large-quantity synthesis of the uric acid is promoted under the action of xanthine oxidase. The kidney is an important excretion organ of uric acid, and can effectively excrete 70% of uric acid produced by normal people every day, and residual uric acid is excreted through intestinal tracts and biliary tracts. The uric acid excretion process in the kidney mainly includes the filtration of glomeruli, reabsorption of renal tubules and collecting ducts, secretion of renal tubules and reabsorption of uric acid after secretion of renal tubules. When the uric acid is produced excessively and exceeds the maximum excretion amount of the human body every day, the renal tubules can play a role in reabsorbing the excessive uric acid, so that the urate is crystallized in the renal tissues, and the absorbed uric acid is transported to the whole body of the human body through blood. In the research, the hesperetin can obviously reduce the serum uric acid level of athletes, and flavonoid compounds such as hesperetin and the like can inhibit xanthine oxidase activity, block a uric acid synthesis way and reduce uric acid generation in combination with literature reports. Meanwhile, the serum urea level of the tested person is further reduced, so that the use of the hesperetin supplement can reduce the production of uric acid and promote the excretion of substances such as uric acid and urea, thereby realizing the function of reducing uric acid.

Serum creatinine and urea are also important indicators for kidney and liver health. Serum creatinine is mainly used to indicate the filtering capability of glomeruli, and an increase in serum creatinine content indicates a decrease in the filtering capability of glomeruli, which is generally caused by diseases such as chronic renal failure. Diseases such as nephritis and chronic renal failure may cause elevation of serum urea, and insufficient protein intake and hepatic failure may cause lower urea. Creatine kinase, found primarily in skeletal muscle, brain and cardiac muscle tissue. Normally, the vast majority of creatine kinase is located in muscle cells, and elevated blood levels of creatine kinase generally indicate that muscle damage has occurred or is occurring. The serum creatinine, urea and creatine kinase levels in this experiment (fig. 5) were significantly lower than those in the control group. These results suggest that hesperetin not only reduces the blood uric acid level and inhibits the secretion of cortisol, but also promotes the glomerular filtration ability and reduces the damage of high-intensity exercise to the liver and kidney. At the end of the study, the inventors found that at the end of the 4 week winter training, both athletes in the control and experimental groups had increased muscle mass, but the experimental group taking the hesperetin supplement (hesperetin 200mg/d, whey protein powder 19.8mg/d) had increased by 0.72% (50.65 + -6.13 kg vs 51.50 + -6.57 kg), while the control group taking the placebo (whey protein powder 20g/d) had increased by only 0.02% (48.01 + -9.13 kg vs 48.14 + -9.17 kg). The inventors believe that this may be attributed primarily to the inhibition of creatine kinase secretion by hesperetin, which reduces muscle damage and thus protects the body.

4. And (4) conclusion:

the inventor mixes the hesperetin with water or separated whey protein, and applies the mixture to the maintenance or the promotion of the body function of an explosive power project athlete during winter training. Research shows that the supplement of the hesperetin (200mg/d of the hesperetin) can reduce the stress response of the body caused by high-intensity exercise during winter training, reduce the high-blood uric acid level caused by exercise, promote the excretion of uric acid, reduce the increase of the contents of urea, creatinine and creatinase in the body after exercise, promote the synthesis of muscles, increase the muscle content and the protein ratio of the body, effectively improve the body components, reduce the stress injury during exercise and improve the adaptability of the body to the high-intensity exercise load. Also, hesperetin can be used as a sports supplement to assist in athlete training, considering that all athletes do not have any physiological or emotional adverse reaction (e.g., insomnia, nausea, irritability, etc.) during taking hesperetin.