Weight reduction application of small molecular cluster water

Abstract

The invention relates to an application of small molecular cluster water in weight reduction, the small molecular cluster water is water with 17O-NMR half peak width less than or equal to 100Hz, preferably less than or equal to 70Hz, the invention also provides an application of the small molecular cluster water in inhibiting weight gain, an application of the small molecular cluster water in reducing the weight of overweight people, an application of the small molecular cluster water in reducing the weight of obese people, and an application of the small molecular cluster water in reducing the weight of the obese people. The traditional Chinese medicine composition can be used for reducing weight of people with overweight hyperuricemia or gout, regulating metabolism and regulating intestinal flora.

Description

Technical Field

[0001] The present invention relates to a new use of small molecule water, and specifically to a weight loss use of small molecule cluster water. Background Art

[0002] In the past 30 years, the prevalence of overweight / obesity in China has increased rapidly. Currently, the overweight / obesity rate of Chinese adults has exceeded 50%, and that of school-age children and adolescents is about 20%. Overweight / obesity will increase the risk of various chronic diseases such as hypertension, T2DM, cardiovascular and cerebrovascular diseases, and specific cancers. Therefore, it is urgent to prevent and control obesity (Zhang Jianduan, et al. 20 Articles on Weight Management for Chinese Residents' Health. Chinese Journal of Diabetes, 2023, 31(12): 881-888.).

[0003] "Report on Nutrition and Chronic Diseases of Chinese Residents (2020)" points out that the problem of overweight and obesity among Chinese residents has become increasingly prominent, and the prevalence / incidence of chronic diseases is still on the rise. The overweight and obesity rates of urban and rural residents in all age groups continue to increase. More than half of the adult residents are overweight or obese, and the overweight and obesity rates of children and adolescents aged 6-17 and under 6 reach 19% and 10.4% respectively.

[0004] Common weight loss methods include limited energy diet, high protein diet, low carbohydrate diet, intermittent energy restriction, low glycemic index diet, weight loss with meal replacement foods, regulation of intestinal microecology, weight loss surgery, exercise, etc. These methods all require deliberate implementation and have low compliance. There is an urgent need for a simple, easy and highly compliant weight loss method.

[0005] Drinking water is a very important health habit, and an appropriate amount of water should be consumed every day. Liquid water in nature does not exist in the form of single molecules (H2O), but in the form of macromolecular cluster water composed of several water molecules, usually more than 10 water molecules. The water molecule clusters aggregated through hydrogen bond interactions are often called "water molecule clusters", in the form of (H2O) nRepresentation. The process in which simple molecules combine into more complex molecular groups without changing the chemical properties of the substance is called molecular association. In the gaseous state, water often exists in the form of single water molecules. However, in the liquid and solid states, single water molecules are unstable and usually multiple water molecules combine to form a relatively stable water molecule cluster. The water in daily life is composed of 15 - 40 water molecules and is often called "large molecular cluster water". For example, purified water is composed of 30 - 40 water molecules. By methods such as applying an external magnetic field, an external electric field, and laser radiation, the hydrogen bonds between water molecules can be broken, the structure of the water molecule cluster can be changed, and the large molecular cluster water can be transformed into small molecular cluster water (n = 2 - 6). When the water molecule cluster becomes smaller, the solubility, permeability, and diffusivity of water are all enhanced, thus having a certain "activation" effect and affecting the physiological functions of organisms in many aspects. Therefore, such water is also called "activated water" (compiled by Wang Min and Yu Chengbao, Water Culture and Water Science, Geological Publishing House, 2022.04). Usually, the 170-NMR half-peak width is measured by a nuclear magnetic resonance spectrometer to characterize the size of the water molecule cluster structure, and the larger its value indicates the larger the cluster structure. There are various methods for preparing small molecular cluster water. For example, CN104230091A discloses a water treatment method for preparing stable small molecular cluster water, and small molecular water with a nuclear magnetic half-peak width value of 47.3 Hz is prepared. Another example is that CN104229925A discloses a water molecule cluster cutting reaction device, and small molecular cluster water with 48.05 Hz and 75.05 Hz is prepared. Tourmaline ultrafine powder has an obvious effect on water molecule clusters. After temperature rise and fall treatment, tourmaline ultrafine powder can make the 17 O-NMR half-height width of water decrease significantly, from 145.41 Hz to 81.11 Hz, that is, the water molecule cluster becomes smaller (Zhan Jie, Wang Zeyan, Wang Peng, etc. Influence of natural mineral tourmaline on water molecule clusters [J]. Journal of Functional Materials, 2010, 41(01): 159 - 161.).

[0006] However, there is little research on the impact of small molecular cluster water on health. Summary of the Invention

[0007] The purpose of the present invention is to provide a new use of small molecular cluster water in weight management.

[0008] The small molecular cluster water is 17 Water with an O-NMR half-peak width ≤ 100 Hz, preferably water ≤ 70 Hz. <{

[0009] Specifically, the present invention provides the following technical solutions:

[0010] The present invention provides the application of small molecular cluster water in weight loss.

[0011] The present invention provides the use of small-molecule cluster water in inhibiting weight gain.

[0012] The present invention provides the use of small-molecule cluster water in reducing the weight of overweight individuals.

[0013] The present invention provides the use of small-molecule cluster water in reducing the weight of obese individuals.

[0014] The present invention provides the use of small-molecule cluster water in reducing the weight of overweight individuals with hyperuricemia or gout.

[0015] The present invention provides the use of small-molecule cluster water in regulating metabolism.

[0016] The present invention provides the use of small-molecule cluster water in regulating the gut microbiota.

[0017] The present invention also provides the use of small-molecule cluster water in the preparation of products for weight loss or inhibition of weight gain; and in the preparation of products for weight management, prevention or treatment of obesity.

[0018] The small-molecule cluster water may further contain excipients permitted in the food or pharmaceutical fields.

[0019] Preferably, small-molecule cluster water is continuously orally administered for at least 3 months.

[0020] Whether the individual has a normal weight or is overweight or obese can be evaluated by the body mass index of the recipient. Body mass index (BMI), also known as the physical fitness index, is an index for calculating weight for height, i.e., BMI = weight / height 2 (kg / m 2 ) (the weight divided by the square of the height, with the weight in kg and the height in m). A BMI between 24.0 - 27.9 indicates overweight, and a BMI ≥ 28 indicates obesity.

[0021] Hyperuricemia refers to a situation where, under normal diet conditions, the fasting blood uric acid level is detected twice at different times, and for men > 420 μmol / L (7 mg / dl) and for women > 360 μmol / L (6 mg / dl).

[0022] In some embodiments of the present invention, the recipient is a healthy human or non-human mammal or a human or non-human mammal that has not developed obesity. In this case, the above applications are for non-therapeutic purposes.

[0023] The beneficial effects of the present invention at least include: The present invention discovers a new function of small molecule cluster water in weight management, and at the same time discovers that small molecule cluster water can regulate the intestinal flora, especially applicable to people with hyperuricemia and gout. Taking small molecule cluster water can achieve an obvious weight loss effect, and the taking method is simple and easy, with high compliance. Weight loss can also reduce the uric acid level of obese gout patients and reduce the occurrence of gouty arthritis.

[0024] Glossary

[0025] Small molecule cluster water: A water molecule cluster composed of 2 to 6 water molecules, with high osmotic pressure, high diffusion ability, high solubility, high oxygen content, and weak alkalinity.

[0026] Regulating metabolism: Refers to energy metabolism. Energy metabolism depends on the presence of various micronutrients, which play the role of precursors, coenzymes or essential components at each cell stage. Most enzymes are regulated by organic cofactors and coenzymes, which act as intermediate carriers for functional groups or electrons transmitted in metabolic reactions. Coenzymes and cofactors play an indispensable role in many cellular metabolic reactions including lipid metabolism and amino acid metabolism.

[0027] Regulating the intestinal flora: The increase of beneficial flora in the intestine and the decrease of pathogenic flora, which can also be said as the increase of the activity of beneficial flora and the decrease of the activity of pathogenic bacteria.

[0028] Product: A product that meets people's needs.

[0029] Food: Finished products and raw materials for human or animal consumption or drinking, as well as items that are both food and traditional Chinese medicine according to tradition, but do not include items for therapeutic purposes.

[0030] Drug: A substance that regulates the physiological functions of humans and specifies indications, functions, main treatments, usage and dosage, including traditional Chinese medicine, chemical drugs, biological products, etc.

[0031] KO or (Uox-KO), English for urate oxidase gene knockout, Chinese for uric acid oxidase gene knockout mice. Description of the Drawings

[0032] Figure 1 Daily water intake change of mice during the experiment

[0033] Figure 2 Body weight change of mice during the experiment

[0034] Figure 3 Effect of small molecule cluster water on intestinal microbial composition

[0035] cel represents the treatment group (small molecule cluster water group), and con represents the control group (purified water group). (A) OTUs analysis of operational taxonomic units (OTUs) isolated from the small molecule cluster water group (red) to control abundance (green). (B) Wilcoxon rank sum test bar chart showing the significance of differences between the small molecule cluster water group and the control group at the genus level. *P < 0.05, **P < 0.01. (C) Microbial composition at the genus level. (D) Branching analysis of the size of the different gut microbiota between the small molecule cluster water group and the control group from phylum to genus. The circles radiating from the inside to the outside represent the taxonomic level from phylum to genus. The diameter of the small circle is positively correlated with the relative abundance.

[0036] Figure 4 Effects of small molecule cluster water on body weight DETAILED DESCRIPTION

[0037] Example 1

[0038] During the study of the effect of small molecule cluster water on gout, the present invention unexpectedly discovered the uric acid-lowering effect of small molecule cluster water and its improvement on metabolic syndrome and renal function in mice. Uox gene knockout spontaneously hyperuricemia mice were used as the test animal model. During the experiment, the animals were allowed to drink the test water freely without restriction. The animals were observed and tested for 3 months. The weight of the mice in the small molecule cluster water group was significantly lower than that in the other groups.

[0039] 1. Test and control substances

[0040] Test sample: Small molecule cluster water (C-cell vitality water from Jilin Tasly Mineral Spring Beverage Co., Ltd., potassium 3.3 mg / L; sodium 16.6 mg / L; calcium 9.9 mg / L; magnesium 8.1 mg / L, half-peak width 48.58 Hz)

[0041] Reference substances: purified water (prepared by a laboratory water purifier), Changbai Mountain mineral water (Jilin Tasly Mineral Spring Beverage Co., Ltd., half-peak width 106.33 Hz)

[0042] 2. Experimental Design and Plan

[0043] 2.1 Mouse selection and grouping

[0044] Uox gene knockout spontaneously hyperuricemic mice (half male and half female): 20-25g, 8-12 weeks old wild-type mice, C57BL / 6J mice, SPF grade (half male and half female): 23-28g, 8-12 weeks old animals are grouped as shown in Table 1:

[0045] Table 1 Grouping of experimental animals

[0046]

[0047] Note: Uox refers to uricase gene knockout mice, which are a spontaneous hyperuricemia mouse model.

[0048] For mice aged 2 - 4 weeks, toe clipping was performed for numbering. Meanwhile, a 3 - 5 mm mouse tail tip was cut and placed into a 1.5 ml EP tube. 40 μL of 50 mmol / L NaOH solution was added (ensuring that the mouse tail was immersed in the NaOH solution), and it was heated and lysed at 99 °C for 20 min. After cooling, 40 μL of 1 mol / L Tris - HCL buffer (pH = 7.5) was added, mixed well, centrifuged at 12000 rpm / min for 10 min at 4 °C, and the supernatant was taken to obtain the DNA stock solution. PCR amplification was performed on the DNA sample to obtain the amplification product, 3% agarose gel electrophoresis was carried out, the characteristics of the bands were observed with a gel imager, and the genotypes of the corresponding mice were recorded. After the animal genotypes were identified, they were raised to 6 - 8 weeks old, and the built - in grouping module of Provantis was used to randomly group them according to body weight and uric acid level. When randomly grouping, 36 Uox gene - knockout hyperuricemia mice screened out were assigned to 3 groups. The difference in body weight of the animals used for grouping should be within ±20% of the average body weight of the same sex. After grouping, the average body weight of each group of animals had no statistical difference at the 5.0% test level.

[0049] 2.2 Experimental protocol

[0050] 2.2.1 Unlimited free drinking water.

[0051] 2.2.2 Experimental period

[0052] The research and observation time lasted for 90 days. During this period, relevant detections were carried out on D1, D7, D14, D21, D28, D42, D56, and D91 days after drug administration.

[0053] 2.2.3 General observation and water intake

[0054] All animals were observed once a day (observed for 3 days before grouping, including the day of grouping). Before and after drug administration on D1, the water intake of the animals was closely observed, and then the water intake was observed and recorded every day.

[0055] 2.2.4 Body weight

[0056] All animals were weighed once before grouping, that is, the grouping body weight (D - 1), and then weighed once on D1, D7, D14, D21, D28, D42, D56, and D91 respectively.

[0057] 3. Experimental results

[0058] 3.1 Changes in water intake and body weight

[0059] During the experiment, the daily water intake of mice was measured. The average 24-hour water intake of KO mice was 7.4 mL, and that of WT mice was 4.7 mL. (See Table 2, Figure 1 ).

[0060] Table 2 Average 24-hour water intake of mice

[0061]

[0062] With age, the body weights of mice in each group showed a gradually increasing trend. The body weights of male KO mice in the group drinking small-molecule cluster water were significantly lower than those in Control Group 1 (KO mice drinking pure water) at D77 and D91 (P < 0.05) ( Figure 2 left); the body weights of female mice in the group drinking small-molecule cluster water were significantly lower than those in Control Group 1 (KO mice drinking pure water) from D35 to D91 (P < 0.05) ( Figure 2 right). This indicates that small-molecule cluster water can inhibit the increase in body weight of mice, reduce the body weight of mice, especially that of hyperuricemia mice.

[0063] Example 2

[0064] This study aimed to evaluate the effect of small-molecule cluster water on the intestinal flora of male hyperuricemia mice. A 13-week drinking water intervention experiment was conducted on Uox gene knockout mice (KO), and the intestinal flora of KO mice was detected by metagenomic sequencing. Intestinal microbiota analysis showed that the activity of beneficial flora related to obesity such as Blautia increased, and the activity of pathogenic bacteria such as Parasutterella decreased. In summary, the intake of small-molecule cluster water can regulate the intestinal flora and reduce the obesity risk caused by hyperuricemia.

[0065] Literature reference for flora:

[0066] 1) HOSOMI, K., SAITO, M., PARK, J., MURAKAMI, H., SHIBATA, N., ANDO, M., NAGATAKE, T., KONISHI, K., OHNO, H., TANISAWA, K., MOHSEN, A., CHEN, Y.A., KAWASHIMA, H., NATSUME-KITATANI, Y., OKA, Y., SHIMIZU, H., FURUTA, M., TOJIMA, Y., SAWANE, K., SAIKA, A., KONDO, S., YONEJIMA, Y., TAKEYAMA, H., MATSUTANI, A., MIZUGUCHI, K., MIYACHI, M. & KUNISAWA, J. 2022. Oral administration of Blautia wexlerae ameliorates obesity and type 2 diabetes via metabolic remodeling of the gut microbiota. Nat Commun, 13, 4477.

[0067] 2) HENNEKE, L., SCHLICHT, K., ANDREANI, N.A., HOLLSTEIN, T., DEMETROWITSCH, T., KNAPPE, C., HARTMANN, K., JENSEN-KROLL, J., ROHMANN, N., POHLSCHNEIDER, D., GEISLER, C., SCHULTE, D.M., SETTGAST, U., TURK, K., ZIMMERMANN, J., KALETA, C., BAINES, J.F., SHEARER, J., SHAH, S., SHEN-TU, G., SCHWARZ, K., FRANKE, A., SCHREIBER, S. & LAUDES, M. 2022. A dietary carbohydrate-gut Parasutterella-human fatty acid biosynthesis metabolic axis in obesity and type 2 diabetes. Gut Microbes, 14, 2057778.

[0068] 1. Materials and methods

[0069] Treatment group: small molecule cluster water (same as Example 1)

[0070] Reference substance: purified water (same as in Example 1)

[0071] The properties of small molecule cluster water are shown in Table 3.

[0072] Table 3 Chemical and physical properties of small molecule cluster water

[0073]

[0074] 2. Animals and experimental design

[0075] Animal experiments followed the principles of experimental animal care and were approved by the Animal Ethics and Welfare Committee of the Affiliated Hospital of Qingdao University. Male C57BL / 6 uox gene knockout mice (KO) at 8 weeks of age with an average body weight of 20 ± 2 g were initially housed in animal cages at a specific pathogen-free level for 14 days. After acclimation, mice with normal body weight were randomly divided into 2 groups (6 mice per group): a control group (KO mice supplemented with purified water) and a treatment group (KO mice supplemented with small molecule cluster water). The experiment was conducted for 91 days, with the water changed every 24 h, the daily water intake measured, and the intestinal contents of the mice collected at the end of the experiment to evaluate the intestinal microbiota (replicate number = 6 / group).

[0076] 3. Metagenomic sequencing and intestinal microbiota analysis

[0077] Use E.Z.N.A. Soil DNA Kit (Omega Bio-tek, GA, USA) was used to extract total genomic DNA from the intestinal contents (10 mg) of mice. Library preparation (Agilent 2100 / Q-PCR) was performed on the amplified DNA, and then sequencing was carried out on the Illumina NovaSeq platform (PE 150, Majorbio Biotechnology Co., Ltd., Shanghai, China).

[0078] 4. Drinking water changes the composition and function of the intestinal flora in KO mice

[0079] To study the effect of drinking small molecule cluster water on the intestinal microbiota of KO mice, metagenomic sequencing was performed. A total of 554 taxonomic units were detected, including 5 domains, 14 kingdoms, 200 phyla, 341 classes, 600 orders, 1044 families, 2978 genera, and 13089 species. We performed principal component analysis (PCoA) on the unweighted distances between groups to determine the effect of different waters on the microbiota. On the first principal coordinate (x-axis), there was a clear separation between the communities, which could explain 71.89% of the variation. The second principal coordinate (y-axis) explained 13.84% of the variance and separated the communities ( Figure 3-A). Analysis at the genus level showed that in mice drinking small-molecule clustered water (cel), it was associated with a decrease in Muribaculaceae and Bacteroidetes, while an increase in Lachnospiraceae ( Figure 3 -C). To evaluate the changes in the microbiota of each group, linear discriminant analysis effect size (LEfSe) analysis was performed to identify the dominant microorganisms in each group. The results showed that compared with the control group, significant changes occurred in the genera Natronorubrum, Natrinema, Methanomicrobium, Candidatus_Methanoplasma, and Cuniculiplasma in the treatment group ( Figure 3 -D). Comparative analysis of the two groups found that in the small-molecule clustered water group, the numbers of beneficial bacteria genera such as Lachnospiraceae, Desulfovibrio, Pseudoflavonifractor, Kazachstania, Dorea, Blautia, Anaerotruncus, and Flavonifractor increased significantly, while the activity of pathogenic bacteria such as the genus Parasutterella decreased ( Figure 3 -B). The experiment demonstrated that small-molecule clustered water can regulate the intestinal flora and enhance the activity of beneficial bacteria.

[0080] Example 3

[0081] This experimental study investigated the effect of small-molecule clustered water on body weight.

[0082] 1. Materials and Methods

[0083] 1.1 Materials

[0084] Specification Purified water (provided by Jilin Tasly Mineral Beverage Co., Ltd.) 330 ml / bottle * 24 bottles / box Small molecule cluster water (C-Vitality Water of Jilin Tasly Mineral Beverage Co., Ltd.) 330 ml / bottle * 24 bottles / box Febuxostat (Jiangsu Wanbang Biochemical Pharmaceutical Co., Ltd.) 40 mg / tablet * 8 tablets * 3 plates Etoricoxib tablets (Hangzhou Merck Sharp & Dohme Pharmaceutical Co., Ltd.) 60 mg / tablet * 5 tablets

[0085] The test medications were provided with unified packaging and labels according to the requirements of the double-blind and double-simulation tests.

[0086] 1.2 Methods

[0087] 1.2.1 Statistical Methods

[0088] Data analysis was performed using SPSS 25.0 (IBM SPSS, Chicago, USA). Measurement data were expressed as (x±s) (normal distribution) or median (interquartile range) (skewed distribution), and count data were expressed as the number of cases. Baseline comparisons were made using independent sample t-tests or Mann-Whitney rank sum tests, and categorical data comparisons were made using chi-square tests. P<0.05 was considered statistically significant, and two-sided tests were used in all cases.

[0089] 1.2.2 Main software systems

[0090] The main computerized systems used in this experiment include, but are not limited to:

[0091] Software name Version number Usage Microsoft Excel 2003、2007 Data entry, calculation, analysis Gout Disease Intelligent Clinic Management System V6.6.210608 Clinical data management IBM SPSS Statistics V25 Test data statistics GraphPad Prism V8 Test data plotting, statistics

[0092] 2. Experimental design and protocol

[0093] 2.1 Experimental design

[0094] This experiment is a single - center, randomized, double - blind, controlled study with an experimental period of 24 weeks.

[0095] Subject population: Patients with primary gout and hyperuricemia.

[0096] 2.2 Number of cases

[0097] According to the blood uric acid level (SU: 420 - 540 and > 540 μmol / L) and body mass index (BMI: 24 - 28 and > 28 kg / m 2 ) After stratification, according to the 1:1 principle, subjects were randomly assigned to group A / B. It was planned to recruit 224 people, and 229 people were actually recruited. 219 people were actually completed, 10 people dropped out, and the dropout rate was 4.37%. 229 subjects were included, including 114 people in group A and 115 people in group B.

[0098] 2.3 Experimental grouping and treatment protocol

[0099] The initial dose of febuxostat is 20 mg qd. If the blood uric acid level does not reach the standard (> 360 μmol / L) after two consecutive follow - ups, the dosing dose of febuxostat is adjusted to 40 mg qd until the end of the trial.

[0100] If a patient has an acute gout attack during the treatment of reducing blood uric acid, etoricoxib 120 mg qd is given for 3 - 5 consecutive days. The treatment protocol is as follows:

[0101] (1) Febuxostat + purified water: Febuxostat 20 / 40 mg qd + (etoricoxib 120 mg qd) + purified water 660 ml tid;

[0102] (2) Febuxostat + small - molecule clustered water group: Febuxostat 20 / forty mg qd + (etoricoxib 120 mg qd) + small - molecule clustered water 660 ml tid;

[0103] 2.4 Randomization and blinding principles:

[0104] A stratified random design was used for randomization grouping. The blind codes for unblinding - group A and group B, experimental group A (febuxostat + purified water), experimental group B (febuxostat + small - molecule clustered water).

[0105] 3. Analysis and Conclusions

[0106] During the 24-week follow-up, the body fat in Group B showed a decreasing trend compared with that in Group A. According to the subgroup analysis results, the decreasing trend of body fat in Group B compared with that in Group A was mainly reflected in the population with a BMI > 28 kg / m 2 . As Figure 4 shown

Claims

1. Use of small molecule cluster water in the preparation of products for reducing body weight or inhibiting body weight gain, wherein the small molecule cluster water is 17 Water with an O-NMR half-peak width ≤ 100 Hz.

2. The application according to claim 1, wherein the small-molecule cluster water is 17 water with an O-NMR half-height width ≤ 70 Hz.

3. The application according to claim 1, wherein the application includes an application in reducing the weight of an overweight person.

4. The application according to claim 1, wherein the application includes an application in reducing the weight of an obese person.

5. The application according to claim 1, wherein the application includes an application in reducing the weight of an overweight person with hyperuricemia or gout.

6. The application according to claim 1, wherein the application includes an application in regulating metabolism.

7. The application according to claim 1, wherein the application includes an application in regulating the intestinal flora.

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