An active oligopeptide RYIVPL, its preparation method and application

Abstract

The present invention relates to an active oligopeptide RYIVPL, its preparation method and application. The active oligopeptide is RYIVPL, and its amino acid sequence is: Arg-Tyr-Ile-Val-Pro-Leu, which has the remarkable function of stimulating intestinal endocrine cells to secrete cholecystokinin (CCK). Compared with the prior art, the oligopeptide of the present invention can be artificially synthesized by a chemical solid-phase synthesis method, or can be obtained by enzymatic hydrolysis of wheat protein followed by separation and purification. The oligopeptide of the present invention has the characteristics of being safe, non-toxic and having no side effects, not being easily enzymolyzed by gastrointestinal digestive enzymes, and being easily absorbed, and is suitable as a functional component for developing drugs with the functions of delaying gastric emptying, promoting satiety and weight loss, and has great application value and broad prospects in the fields of medicine and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of bioactive peptides, and particularly relates to an active oligopeptide RYIVPL, a preparation method thereof, and an application thereof. Background Art

[0002] Obesity has become a global public health problem. Many chronic diseases, including atherosclerosis, cardiovascular diseases, type II diabetes, etc., are related to obesity. The imbalance between energy intake and energy expenditure is the direct cause of individual overweight and obesity. Therefore, in order to prevent obesity, restricting excessive energy intake, that is, restricting food intake, is the best means. The hypothalamus of the body's feeding center and the digestive tract are the key parts of the body to control feeding regulation, and many brain-gut peptides related to feeding regulation are distributed therein. Cholecystokinin (CCK) is a member of the classical brain-gut peptides, which can regulate the body to produce satiety signals, reduce the body's food intake, and thus play a role in suppressing appetite. Existing studies have shown that the secretion of CCK is regulated by dietary factors. Therefore, it is possible to improve or relieve obesity and related chronic diseases by dietary regulation of intestinal CCK secretion.

[0003] Food-derived bioactive peptides are a common dietary factor, which have the characteristics of being easily digested and absorbed by the human body and high food safety, and have a wide range of applications in the food field. In view of the important role of CCK and the many advantages of bioactive peptides, it is of great value to find a bioactive peptide or extract that can promote intestinal CCK secretion, and it has great economic and social significance for preventing obesity or related chronic diseases. Summary of the Invention

[0004] Based on the current situation that there is a lack of bioactive peptides that can promote intestinal CCK secretion in the prior art, the present invention provides an active oligopeptide RYIVPL with the function of promoting CCK secretion, a preparation method thereof, and an application thereof.

[0005] The object of the present invention can be achieved by the following technical solutions:

[0006] In the first aspect of the present invention, an active oligopeptide with CCK secretion-promoting activity is provided, and the active oligopeptide is RYIVPL, and the amino acid sequence is: Arg-Tyr-Ile-Val-Pro-Leu.

[0007] Wheat protein is mainly gluten and gliadin. It is found that the RYIVPL active oligopeptide exists in wheat protein and is derived from wheat Protein kinase domain-containing protein (Uniprot protein accession number: A0A3B6D6B8). Therefore, the RYIVPL active oligopeptide can be prepared from wheat protein.

[0008] In the second aspect of the present invention, there is provided a polynucleotide encoding the active oligopeptide.

[0009] In the third aspect of the present invention, there is provided a method for preparing the active oligopeptide, which is obtained by subjecting wheat protein to biological enzymatic hydrolysis followed by separation and purification, or directly obtained by artificial synthesis through genetic engineering methods, or directly prepared by chemical synthesis.

[0010] Artificial synthesis of the active oligopeptide by genetic engineering methods is a technical solution that can be achieved by those skilled in the art. For example, it can be based on DNA recombinant technology and control the sequence synthesis of the oligopeptide through a suitable DNA template.

[0011] The method of directly obtaining from wheat protein through separation and purification can be: based on the amino acid sequence of the given active oligopeptide, using conventional enzymatic hydrolysis, separation, and purification methods in biological technology to obtain the active oligopeptide from wheat.

[0012] The method of preparing by chemical synthesis is to synthesize the above active oligopeptide by using the traditional solid-phase synthesis method.

[0013] In the fourth aspect of the present invention, there is provided a method for preparing an enzymatic hydrolysis product containing the active oligopeptide, which enzymatically hydrolyzes wheat protein by a two-step enzymatic hydrolysis method, including the following steps: enzymatically hydrolyzing wheat protein with pepsin and trypsin in sequence to obtain a wheat protein enzymatic hydrolysis product, which is the enzymatic hydrolysis product containing the active oligopeptide.

[0014] In an embodiment of the present invention, the method of enzymatically hydrolyzing wheat protein with pepsin and trypsin in sequence includes the following steps:

[0015] 1) Extract wheat protein from wheat;

[0016] 2) Enzymatically hydrolyze the wheat protein with the pepsin to obtain a first enzymatic hydrolysis product;

[0017] 3) Enzymatically hydrolyze the first enzymatic hydrolysis product with the trypsin to obtain a wheat protein enzymatic hydrolysis product.

[0018] In an embodiment of the present invention, the method for extracting wheat protein from wheat includes the following steps:

[0019] Grind wheat into flour, degrease it, then add it to distilled water, set the temperature to 50 - 60 °C, adjust the pH to 4 - 6, and pretreat it with glucoamylase for 0.5 - 2 h; after cooling to room temperature, adjust the pH to 10 - 12 for protein extraction, and after centrifugation, take the supernatant, adjust the pH of the supernatant to 4.4 - 4.6, let it stand and then centrifuge, wash with water; finally dry to obtain wheat protein.

[0020] In one embodiment of the present invention, the mass ratio of the pepsin or trypsin to the wheat protein is 1:10 - 100.

[0021] In one embodiment of the present invention, the enzymatic hydrolysis conditions of the pepsin or trypsin are enzymatic hydrolysis at 37°C for 1 - 4 h.

[0022] In one embodiment of the present invention, after obtaining the wheat protein hydrolysate in step 3), the following steps are further included:

[0023] The wheat protein hydrolysate is separated using a YMC ODS C18 chromatographic column, and 5% methanol deionized water solution, 30% methanol deionized water solution, 80% methanol deionized water solution, and 100% methanol solution are used as eluents, and elution, chromatography purification are carried out at a certain flow rate, and different components are collected; then the effects of different components on the secretion of cholecystokinin (CCK) by enteroendocrine cells are detected, and the component with the highest activity of stimulating CCK secretion is selected as the target component, that is, the product containing the active oligopeptide.

[0024] In the fifth aspect of the present invention, there is provided an enzymatic hydrolysate of an active oligopeptide prepared based on the above preparation method.

[0025] In the sixth aspect of the present invention, there is provided the application of the active oligopeptide and the enzymatic hydrolysate containing the oligopeptide in the preparation of a product having at least one of the following functions 1) - 4):

[0026] 1) Promote CCK secretion;

[0027] 2) Slow down gastric emptying;

[0028] 3) Inhibit appetite and reduce food intake;

[0029] 4) Prevent or assist in the treatment of obesity.

[0030] The active oligopeptide and the enzymatic hydrolysate containing the active oligopeptide have the function of promoting CCK secretion, and can slow down gastric emptying, inhibit appetite and reduce food intake, and prevent or assist in the treatment of obesity.

[0031] In the seventh aspect of the present invention, there is provided a product, which contains the active oligopeptide or the enzymatic hydrolysate containing the active oligopeptide, and the product has at least one of the following functions 1) - 4):

[0032] 1) Promote CCK secretion;

[0033] 2) Slow down gastric emptying;

[0034] 3) Inhibit appetite and reduce food intake;

[0035] 4) Prevent or assist in the treatment of obesity.

[0036] The product described above includes drugs.

[0037] Compared with the prior art, the advantages and beneficial effects of the present invention are embodied in the following aspects:

[0038] The present invention has screened out active oligopeptides with the ability to promote intestinal CCK secretion. These active peptides have a novel peptide sequence structure and there have been no other related reports so far. The active peptides described in the present invention can be prepared from food protein wheat protein and have the advantages of being safe, non-toxic and having no side effects. The active oligopeptides are not easily degraded by pepsin and trypsin in the gastrointestinal tract and have good stability, thus maximizing the CCK secretion-promoting activity. The active oligopeptides described in the present invention have a molecular weight of less than 1000 Da. With a small molecular weight, in addition to promoting intestinal CCK secretion activity, they are also easily absorbed by the body and have good nutritional functions. The preparation method of the active oligopeptides provided by the present invention or the enzymatic hydrolysate containing the oligopeptides is simple, easy to operate, convenient for large-scale industrial production, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 Shows the effect of RYIVPL active oligopeptide on the viability of STC-1 cells;

[0040] Figure 2 Shows the effect of RYIVPL active oligopeptide on the secretion of CCK by STC-1 cells;

[0041] Figure 3 Shows the effect of wheat protein hydrolysate (W) on the secretion of CCK by STC-1 cells, with C being the control group (without wheat protein hydrolysate);

[0042] Figure 4 Shows the effect of wheat protein hydrolysate on the secretion of CCK by mouse intestinal endocrine cells;

[0043] Figure 5 Shows the chromatogram of wheat protein hydrolysate separated by YMC ODS C18 column;

[0044] Figure 6 Shows the effect of the separated components on the secretion of CCK by STC-1 cells;

[0045] Figure 7 Shows the MS / MS spectrum and sequence analysis of the RYIVPL active oligopeptide in the separated component F7. DETAILED DESCRIPTION OF THE INVENTION

[0046] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.

[0047] Example 1 Artificial synthesis of RYIVPL active oligopeptide and evaluation of its CCK secretion-promoting activity

[0048] I. Synthesis of RYIVPL Active Oligopeptide

[0049] The active oligopeptide Arg-Tyr-Ile-Val-Pro-Leu (RYIVPL) was synthesized by "Zhejiang Hongtuo Technology Co., Ltd." using the peptide solid-phase synthesis method, and the purity of the synthesized peptide was verified to be greater than 95% by high-performance liquid chromatography method and mass spectrometry technology.

[0050] II. Effects of RYIVPL on STC-1 Cell Activity and CCK Secretion

[0051] (1) Culture of STC-1 Cells

[0052] STC-1 cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS), 1% non-essential amino acids (NEAA), 100 U / mL penicillin and 0.1 mg / mL streptomycin. The cells were incubated in a cell culture incubator at 37°C with 5% CO2, and when they reached 80-90% density, they were passaged by trypsin digestion.

[0053] (2) Determination of Cell Activity

[0054] The effects of the RYIVPL active peptide on the viability of STC-1 cells were tested and evaluated by WST-8 (2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium monosodium salt) cell proliferation and cytotoxicity assay. WST-8 was added to the STC-1 cells treated in a 96-well plate. In the presence of an electron coupling reagent, WST-8 can be reduced by some dehydrogenases in the mitochondria to produce orange-yellow Formazan, and the absorbance was measured at a detection wavelength of 450 nm with an enzyme-linked immunosorbent assay reader, and the results were expressed as a percentage of the control group. The results are as Figure 1 shown. At different test concentrations, the viability of STC-1 cells did not change compared with the control group, indicating that the RYIVPL active peptide is non-toxic to cells.

[0055] (3) Determination of Hormone Content Secreted by STC-1 Cells

[0056] The RYIVPL active oligopeptide was made into peptide solutions with molar concentrations of 0.2, 2, and 5 mM respectively using Hank's buffer. STC-1 cells were seeded in a 24-well culture plate at a density of 1.25×10 5 cells. When the cells reached 80%-90% confluence, the cells were washed twice with Hank's buffer to remove the culture medium. The peptide solution was added to the STC-1 cells, and the cells were incubated in an incubator at 37°C for 2 h. After incubation, the cells were centrifuged at 1000 g for 20 min, and the supernatant was taken. The CCK content was measured using a commercial CCK kit from Wuhan Yunke Long Co., Ltd.

[0057] Taking the total protein content of the supernatant measured by the BCA kit of Beyotime Biotechnology Co., Ltd. as a reference to reduce the error caused by operations between different batches. The calculation formula is as follows: C CCK / (C BCA ×100)(×10 -7 ), where C CCK represents the CCK concentration (pg / mL) in the supernatant, and C BCA represents the total protein concentration measured in the supernatant minus the measured peptide concentration (mg / mL).

[0058] The effect of RYIVPL bioactive peptide on the secretion of CCK by STC-1 cells is shown in Figure 2 . It can be seen that RYIVPL bioactive oligopeptide dose-dependently increases the secretion of CCK. A large number of studies have confirmed that CCK is an important member of the brain-gut peptides, which can slow down gastric emptying, regulate the body to generate satiety signals, reduce the food intake of the body, and thus play a role in suppressing appetite. Increasing the secretion of intestinal CCK is of great significance for preventing and alleviating obesity. The RYIVPL bioactive peptide described in this patent can significantly promote the secretion of CCK by intestinal endocrine cells STC-1. Therefore, these peptides have important significance and application value for promoting satiety and weight loss.

[0059] Example 2 Preparation of wheat protease hydrolysate and activity evaluation

[0060] (1) Preparation of wheat protease hydrolysate

[0061] Grind wheat into flour and pass it through an 80-mesh sieve, then defat the wheat flour with hexane. Soak the defatted wheat flour in distilled water in a beaker at a mass ratio of 12:1, adjust the pH value to 5.0 with 1 mol / L HCl, and treat it with glucoamylase at 50 °C for 1 h. Then adjust the pH value to 11.0 with 1 mol / L NaOH, centrifuge to obtain the supernatant after stirring with a magnetic stirrer for 2 h. Adjust the pH value of the supernatant to its isoelectric point (pH 4.5) with 1 mol / L HCl, let it stand for 1 h and then centrifuge, wash the precipitate until it is neutral, redissolve it with a small amount of distilled water and then freeze-dry to obtain wheat protein, and store it at 4 °C for later use.

[0062] Dissolve 1 g of freeze-dried protein powder in 20 mL of an aqueous solution containing 25 mg of freshly prepared pepsin, adjust the pH value of the solution to 2.0 with HCl (1 mol / L), and incubate it at 37 °C for 2 h. After the incubation is over, adjust the pH value of the solution to 6.8 with NaOH (1 mol / L), then add 50 mg of trypsin and continue enzymatic hydrolysis for 2 h. After completion, inactivate the enzyme in a boiling water bath for 8 min, centrifuge to obtain the supernatant and freeze-dry it to obtain wheat protease hydrolysate.

[0063] (2) Activity evaluation

[0064] The wheat protease hydrolysate was made into a solution with a mass concentration of 5 mg / mL using Hank's buffer, and the effect of the hydrolysate on the secretion of CCK by STC-1 cells was measured by the above method. The results are shown in Figure 3 . It can be seen from the results that the wheat protease hydrolysate can significantly stimulate the secretion of CCK by STC-1 cells.

[0065] Furthermore, the effect of wheat protease hydrolysate on the secretion of hormones by mouse intestinal endocrine cells was evaluated at the animal level. After a 1-week adaptation period, ICR mice were randomly divided into 2 groups (28 mice in each group). Control group: gavaged with normal saline; Wheat protease hydrolysate group: gavaged with wheat protease hydrolysate (1.0 g / kg body weight). After gavage, blood was collected by eye socket extraction at 0, 15 min, 30 min, 60 min, 90 min, 120 min, and 150 min, and placed in a centrifuge tube containing EDTA (final concentration of 1 mg / mL) and aprotinin (final concentration of 0.6 TIU / mL). The supernatant was obtained by centrifugation, and the content of CCK hormone in the serum was measured by ELISA method. The CCK in the serum of the normal saline gavage group maintained at about 40 pg / mL during this period. The results of the wheat protease hydrolysate gavage group are as Figure 4 shown. It can be found that the wheat protease hydrolysate greatly increases the CCK level in mice. At 60 min, the concentration of CCK in the mouse blood reaches about 90 pg / mL.

[0066] Example 3 Preparation of active oligopeptides promoting CCK secretion from wheat protein

[0067] Take YMC ODS C18 packing material for conventional swelling and column packing (the packing buffer solution is methanol). The column height is 25 cm, the inner diameter is 1.6 cm, and there must be a water layer of 1.5 - 2 cm at the top of the column at any time. After the column is equilibrated with about 3 - 4 column volumes, it can be used. When the liquid level is reserved at 2 - 3 mm, the wheat protease hydrolysate is added for sampling. Weigh about 160 mg of wheat protease hydrolysate powder and dissolve it in 2 mL of distilled water. After filtering through a 0.45 μm microporous filter membrane, it is added to the chromatography column. The eluent is methanol solutions with different volume concentrations (5%, 30%, 80%, 100%), the elution speed is 2 mL / min, and the elution peaks are collected. The separation map of the wheat protease hydrolysate is shown in Figure 5 , and it can be seen that the C18 chromatographic column divides the protease hydrolysate into 7 peptide components.

[0068] The first component has extremely strong water absorption and can absorb the moisture in the air after freeze-drying. In addition, the fourth component accounts for a very small proportion and is extremely difficult to obtain. Therefore, the activity of 5 peptide components was evaluated by the above method, and the results are as Figure 6 shown. It can be found that among the 5 peptide components, the F7 component has the best ability to stimulate the secretion of CCK by STC-1 cells. Therefore, the F7 component is a highly active oligopeptide promoting CCK secretion.

[0069] Example 4 Identification of Wheat Protein Containing RYIVPL Active Peptide

[0070] The peptide sequence in the F7 fraction was identified by mass spectrometry. The sample was dissolved in distilled water to make a 1 mg / mL sample. A reversed-phase chromatography column (150 μm i.d.×150 mm, packed with Acclaim PepMap RPLC C18, 1.9 μm, ) was used for separation. Mobile phase A was 0.1% formic acid aqueous solution, and mobile phase B was 0.1% formic acid / 80% acetonitrile solution. Gradient elution was performed at a flow rate of 600 nL / min. Separation gradient: 0 - 2 min, 4 - 8% B; 2 - 45 min, 8 - 40% B; 45 - 55 min, 40 - 60% B; 55 - 56 min, 60 - 95% B; 56 - 66 min, 95% B. The mass spectrometry ion source type was electrospray ionization source (ESI), positive ion scan mode, spray voltage 2200 V, capillary temperature 270 °C. Primary mass spectrometry parameter settings: scan range 100 - 2000 m / z, maximum resolution 70000, automatic gain parameter 3000000. Secondary mass spectrometry parameter settings: scan range 50 - 2000 m / z, maximum resolution 17500, automatic gain parameter 100000.

[0071] After mass spectrometry detection, the main ion peak in the F7 fraction was m / z = 380.74, z = 2. The molecular ion peak was further analyzed by secondary mass spectrometry. The secondary mass spectrometry diagram of this molecular ion peak is shown in Figure 7 . After database matching, the peptide corresponding to the ion peak was Arg - Tyr - Ile - Val - Pro - Leu (RYIVPL).

[0072] Wheat protein is mainly glutenin and gliadin. It can be found that the RYIVPL active oligopeptide is derived from wheat Protein kinase domain - containing protein (Uniprot protein accession number: A0A3B6D6B8), and the RYIVPL active oligopeptide can be prepared from wheat protein.

[0073] The amino acid sequence of wheat Protein kinase domain - containing protein (Uniprot protein accession number: A0A3B6D6B8) is shown in SEQ ID NO.1. In the present invention, the RYIVPL active oligopeptide is located at positions 602 - 607 of the amino acid sequence shown in SEQ ID NO.1.

[0074] The specific amino acid sequence shown in SEQ ID NO.1 is as follows:

[0075] MHERAVWIFL CDLGGQNMPF EGSIPVEIGN LVNLFSLGME SNFLYGSIPS SIGKLRNLYI 60

[0076] LNLSKNKLSG QIPPSIGDVT QLALLYLDGN NLGGNIPGSL AQCMGLFELN LSQNNLDGSI 120

[0077] PVRLFADLLF AWSLDFSHNN LTGELPPVLG TYGIGSGLVS LHMEGNRFHG QIPERWHLLV 180

[0078] STRQINLSHN DLSGVVPKFF EQLDKLEQLD LSYNNLEGAV PASGIFTHSA AVVLDGNKGL 240

[0079] CSNSPRLALP ICPGISGSAT KVKHHLSLLV TSLLIVLPPL TIGSLVLIWF MLTIWKKGSF 300

[0080] PFSRWDLVSK MFPNRREVHT APCHDEKKLK RVSYEDIVKA TNWFSLVHTI SSTCTGSVYV 360

[0081] GRFKSDKSLV AIKVFKLNEP GGYDSYLIEC EVLRSTRHRN IMRPVTLCST LDSQNHEFKA 420

[0082] LIFEFMVNGS LEAWLHSEQH NGIPDKVLGF GQRICIAADV ASALDYAHNE LTPPLIHCDL 480

[0083] KPNNVLLDDD MTARLSDFGS AKFLSPGLVIPKSLDDVGGT IGYVAPEYGM GCEISVAGDL 540

[0084] YSFGVLLLEL LTGKRPTDDM FVDGLSLCKF SESMFPDRVA EILDPHMAHE EHQGCTEAWM 600

[0085] ERYIVPLVAL GLSCTMESTK DRPGMKDVCA KLSDIRASFL ELS 643

[0086] The above description of the embodiments is to enable those of ordinary skill in the art to understand and use the invention. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative efforts. Therefore, the present invention is not limited to the above embodiments, and all improvements and modifications made by those skilled in the art without departing from the scope of the present invention according to the disclosure of the present invention should be within the protection scope of the present invention.

Claims

1. An active oligopeptide with the function of promoting cholecystokinin secretion, characterized in that, The oligopeptide is RYIVPL, and its amino acid sequence is: Arg-Tyr-Ile-Val-Pro-Leu.

2. A polynucleotide encoding the active oligopeptide according to claim 1.

3. A preparation method of the active oligopeptide according to claim 1, characterized in that, It is directly obtained by chemical synthesis, or artificially synthesized by genetic engineering methods, or obtained from wheat protein through enzymatic hydrolysis and separation and purification methods.

4. A preparation method of an enzymatic hydrolysate containing the active oligopeptide according to claim 1, characterized in that, It includes the following steps: Successively hydrolyze wheat protein with pepsin and trypsin to obtain a wheat protein hydrolysate, which is the hydrolysate containing the active oligopeptide described in claim 1.

5. According to the preparation method described in claim 4, characterized in that, The method for extracting wheat protein from wheat includes the following steps: Grind wheat into flour, degrease it, then add it to distilled water, set the temperature at 50-60 °C, adjust the pH to 4-6, and pretreat it with glucoamylase for 0.5-2 h; after cooling to room temperature, adjust the pH to 10-12 for protein extraction, and after centrifugation, take the supernatant. Adjust the pH of the supernatant to 4.4-4.6, let it stand and then centrifuge, wash with water; finally dry to obtain wheat protein.

6. According to the preparation method described in claim 4, characterized in that, The mass ratio of the pepsin or trypsin to the wheat protein is 1:10-100; The enzymatic hydrolysis condition of the pepsin or trypsin is enzymatic hydrolysis at 37 °C for 1-4 h.

7. According to the preparation method described in claim 4, characterized in that, After obtaining the wheat protein hydrolysate, the following steps are also included: Separate the wheat protein hydrolysate using a YMC ODS C18 chromatographic column, use 5% methanol deionized water solution, 30% methanol deionized water solution, 80% methanol deionized water solution and 100% methanol solution as eluents, and perform elution, chromatography purification at a certain flow rate, and collect different components; then detect the effects of different components on the secretion of cholecystokinin (CCK) by enteroendocrine cells, and select the component with the highest activity of stimulating CCK secretion as the target component, which is the active oligopeptide described in claim 1.

8. An enzymatic hydrolysate containing the active oligopeptide with the function of promoting cholecystokinin secretion according to claim 1, prepared based on the preparation method described in any one of claims 4 - 7.

9. Use of the active oligopeptide according to claim 1 and the enzymatic hydrolysate containing the active oligopeptide according to claim 1 in the preparation of a product having at least one of the following functions: Slowing down gastric emptying; Inhibiting appetite and reducing food intake; Preventing or assisting in the treatment of obesity; The product is a medicine.

10. A product, characterized in that, It contains the active oligopeptide described in claim 1, or the hydrolysate containing the active oligopeptide described in claim 1, and the product has at least one of the following functions: Slow down gastric emptying; Inhibit appetite and reduce food intake; Prevent or assist in the treatment of obesity; The product is a medicine.

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