Application of Sanzan gum in preparation of drugs for reducing blood fat and blood sugar

Abstract

The invention discloses application of Sanzan gum in preparation of drugs for reducing blood fat and blood sugar. Belongs to the technical field of health care. The invention finds that the Sanzan gum can effectively reduce food intake and weight, and can obviously reduce blood sugar and triglyceride. The pharmaceutical composition is high in safety and can be safely applied to the pharmaceutical composition for reducing blood sugar or blood fat.

Description

Technical Field

[0001] The invention relates to the technical field of health care, and more particularly to the application of trisammonia gum in the preparation of drugs for lowering blood lipids and blood sugar. Background Art

[0002] Sanzan gum, also known as sphingosine gum Ss, is an extracellular polysaccharide obtained by the metabolism of Sphingomonas sanxanigenens with glucose and inorganic nitrogen sources as raw materials under sufficient oxygen and aseptic conditions. It can form an elastic gel with a spirally wound structure. Its aqueous solution is a typical pseudoplastic fluid, and the viscosity decreases with the increase of shear rate. When the mass fraction of Sanzan gum solution is ≥0.6% and heated to above 75°C, it can form a stable gel after cooling to room temperature. It was initially mainly used in the materials and petroleum industries.

[0003] On May 20, 2020, the National Health Commission officially issued Announcement No. 4 of 2020, approving Sanzan gum as a food thickener, stabilizer and coagulant for use in meat sausages (food category 08.03.05), fruit and vegetable juice (pulp) beverages (food category 14.02.03) and plant protein beverages (food category 14.03.02) to improve the sensory quality and stability of the products.

[0004] As a newly approved food additive, there are few studies on the properties and applications of Sanzan gum at home and abroad, especially no reports on the lipid-lowering and blood sugar-lowering functions of Sanzan gum.

[0005] In summary, exploring the application of Sanzan gum in lowering blood lipids and blood sugar is an urgent problem that technicians in this field need to solve. Summary of the invention

[0006] In view of this, the present invention provides the use of Sanzan gum in the preparation of lipid-lowering and blood sugar-lowering drugs.

[0007] In order to achieve the above object, the present invention adopts the following technical solution:

[0008] Application of Sanzan gum in the preparation of lipid-lowering and blood sugar-lowering drugs.

[0009] Further, Tri-Gel is used to lower blood sugar and triglyceride levels.

[0010] Further, Tri-Gel is used to reduce food intake.

[0011] Further, Sanzan gum is used to reduce body weight.

[0012] It can be seen from the above technical solution that, compared with the prior art, the beneficial effects achieved by the present invention are:

[0013] The present invention finds that Sanzan gum can effectively reduce food intake, reduce body weight, and has obvious blood sugar and triglyceride lowering effects. It has high safety and can be safely used in combination with blood sugar or blood lipid lowering drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0015] Figure 1 The effect of Sanzan gum in Example 1 of the present invention on the body weight of male high-sugar and high-fat rats;

[0016] Figure 2 This is the effect of Sanzan gum in Example 1 of the present invention on the body weight of female high-sugar and high-fat rats. DETAILED DESCRIPTION

[0017] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0018] The drugs required for the present invention are conventional experimental drugs, which are purchased from commercial channels; the experimental methods not mentioned are conventional experimental methods and will not be described in detail here.

[0019] The following experiment was commissioned by Hebei Fengchuan Biotechnology Co., Ltd. and Hebei Xinhe Biochemical Co., Ltd. to the New Drug Safety Evaluation Research Center of Hebei Medical University. Research topic name: Effects of Sanzan Gum on Glucose and Lipid Metabolism in High-sugar and High-fat Rats. Research topic number: NPJ-202407-PHD1.

[0020] Example 1

[0021] 1 Materials and methods

[0022] 1.1 Experimental Materials

[0023] (1) Test substance information

[0024] The test substance Sanzan Gum was provided by Hebei Fengchuan Biotechnology Co., Ltd., and its main components are composed of sugars, lipids and polypeptides. It is produced by using Sphingomonas sp. strain TP-3 (Sphingomonas sanxanigenens sp.nov.) as the producing bacteria, starch or glucose as the main raw materials, and is produced by biological fermentation, extraction, drying and crushing. Its molecular structure is as follows:

[0025]

[0026] The specific information of Sanzan Glue is as follows:

[0027] Table 1 Information of Sanzan Glue

[0028]

[0029] (2) Solvent reference material information

[0030] Chinese name: sodium carboxymethyl cellulose;

[0031] Batch number: 20230701;

[0032] Molecular formula: (fill in for chemical drugs) or main component: C6H7O2(OH)2OCH2COONa;

[0033] Manufacturer: Tianjin Damao Chemical Reagent Factory;

[0034] Preparation method and concentration: Accurately weigh 5g of sodium carboxymethyl cellulose, pour into a 1L volumetric flask, add about 700ml of ultrapure water, cover the bottle cap, put it into an ultrasonic instrument for 3 hours, cool it down, add ultrapure water to 1L, and make the concentration 0.5%. Put it in a 1L reagent bottle for use (prepare once a week).

[0035] (3) Main reagents

[0036] Streptozotocin (STZ): purchased from Beijing Solaibao Biotechnology Co., Ltd., 1 g / bottle, Lot No. 24240306009, valid until 20260307, ​​stored at -20°C. Prepare a 1% solution with citric acid buffer (pH 4.5) before use, keep in an ice bath away from light, and administer by intraperitoneal injection within 30 minutes.

[0037] 0.1 M sodium citrate buffer (sterile): purchased from Beijing Solebow Biotechnology Co., Ltd., 500 mL / bottle, pH 4.5, Lot No. 240002002, valid until 20250219.

[0038] 1.2 Experimental animals and feeding conditions

[0039] (1) Experimental animals

[0040] Table 2 Experimental animal information

[0041]

[0042] (2) Methods for identifying experimental animals

[0043] After the experimental animals arrived, they were received and quarantined according to the SOP requirements for animal reception and quarantine. According to gender, each animal was assigned an independent animal number (male: M01, M02, M03..., female: F01, F02, F03...). Picric acid was used to mark the animals according to the SOP for animal marking, and the animal number was used for identification in the original data.

[0044] (3) Animal rearing conditions

[0045] During the experiment, the animals were kept in the W6 laboratory at a temperature of 19-26°C, a relative humidity of 40-70%, 12 hours of light and 12 hours of darkness. The rats were kept in a special feeding box, fed regularly and quantitatively, and had free access to food and water.

[0046] Feed: Ordinary feed for mice and high-sugar and high-fat feed (67% rat maintenance feed + 10% lard + 20% sucrose + 2.5% cholesterol + 0.5% sodium cholate), Ke'ao Xieli (Tianjin) Feed Co., Ltd., meeting national standards.

[0047] Drinking water: sterile water, homemade.

[0048] 1.3 Experimental methods

[0049] (1) Basis for dosage design

[0050] The maximum dietary recommended amount of Sanzan Glue is 1.3g / 60kg / d. Combined with the dissolution test results of this laboratory, the test substance was set at three doses, namely the low, medium and high dose groups of Sanzan Glue, and the doses for rats were 0.14, 0.336 and 0.672g / kg, respectively. The above doses are equivalent to 6.5 times, 15.5 times and 31 times the recommended human intake when converted by kilogram body weight; and 1 times, 2.4 times and 4.8 times the recommended human intake when converted by body surface area.

[0051] (2) Dose group design

[0052] Table 3 Dose group design

[0053]

[0054] (3) Experimental animal grouping design

[0055] 44 SD rats that passed the quarantine were selected, half of them were male and half were female. They were adaptively fed for 4 days before the experiment, and then all animals were fasted but not watered overnight. They were randomly divided into 5 groups according to blood sugar and body weight, namely normal control group, model control group, low-dose, medium-dose and high-dose groups of the test substance, with 8 to 10 rats in each group. The details are as follows:

[0056] Table 4 Experimental animal grouping design

[0057]

[0058] (4) Route and method of administration

[0059] Administration route: intragastric administration;

[0060] Dosing volume: 20mL / kg;

[0061] Dosing frequency: once a day;

[0062] Dosing date: 2024-03-19 to 2024-05-13.

[0063] The normal control group was given a standard rat maintenance diet, and the other groups were given a high-sugar and high-fat diet (67% rat maintenance diet + 10% lard + 20% sucrose + 2.5% cholesterol + 0.5% sodium cholate) to prepare a high-sugar and high-fat rat model. Four weeks later, the rats in the four modeling groups were intraperitoneally injected with 1% streptozotocin (STZ) 35 mg / kg at one time. On the third day after the injection of STZ, all animals were fasted overnight but not water, and 0.3-0.4 ml of blood was collected from the inner canthus (procoagulant), and the related indicators of glucose and lipid metabolism (TCHO, TG, LDL-C, HDL-C, GLU) were measured. If necessary, an additional injection of STZ could be given, and blood sugar could be measured once more to determine whether the high-sugar and high-fat model was successful.

[0064] This study adopted a preventive drug administration method, starting from the first day of modeling. All animals were administered by gavage. The low, medium and high dose groups of the test substance were given 0.14, 0.336 and 0.672 g / kg of tri-zan gum suspension, and the normal control group and the model group were given the same volume of 0.5% CMCNa at the same time. The administration volume was 2 mL / 100 g, once a day, for 8 weeks.

[0065] (5) Observation indicators and methods

[0066] Body weight: Animals were weighed twice a week and their body weight changes were recorded.

[0067] Food intake: According to the specific food intake growth, a fixed amount of feed was given to each cage, and the remaining amount was weighed twice a week to calculate the daily intake of each rat.

[0068] Food intake = (fixed food intake - remaining food intake) / number of animals per cage / number of days.

[0069] Blood glucose and blood lipid related indicators: 0.3-0.4 mL of blood was collected from the inner canthus (procoagulant) before grouping, 4 weeks + 3 days after modeling, 5 weeks of drug administration (blood glucose was measured by blood glucose meter), 6 weeks of drug administration, and at the end of 8 weeks of drug administration, and the following blood glucose and blood lipid related indicators were measured.

[0070] Table 5 Blood sugar and blood lipid related indicators

[0071]

[0072] (6) Statistical methods

[0073] All blood biochemistry results were collected automatically and printed out by computer. The rest of the raw data were collected manually. Excel software and GraphPad Instat were used to process and statistically analyze the test data in accordance with the SOP for statistical analysis of test data. GraphPad Prism software was used to draw the test graphs.

[0074] For normally distributed measurement data, if the variance is equal, a "t" test or variance analysis (Dunnet test if there is a significant difference) is performed between each dose group and the control group. If the variance is unequal, a "t'" test is performed between each dose group and the control group. For non-normally distributed measurement data and measurement data with unequal variance after data transformation, non-parametric tests such as rank sum test are used. For count data, chi-square test is performed.

[0075] 2 Experimental results

[0076] (1) Observation of general symptoms

[0077] Before administration, the animals in each group developed normally, had shiny fur, normal food and water intake, no loose stools, no abnormal secretions, and normal activities. During the experiment, the weight of the rats in the normal control group increased steadily, the fur was shiny, and the activities were agile. The fur of the rats in the modeling groups was rough, and no other obvious abnormalities were found.

[0078] (2) Impact on body weight

[0079] like Figure 1 , Figure 2 , as shown in Table 6 and Table 7.

[0080] Table 6 Effect of Sanzan Glue on Body Weight of Male High-sugar and High-fat Rats (g, Mean±SD)

[0081]

[0082]

[0083] Note: Compared with the normal control group, *P<0.05, **P<0.01.

[0084] Table 7 Effect of Sanzan Glue on Body Weight of Female High-sugar and High-fat Rats (g, Mean±SD)

[0085]

[0086]

[0087] Note: Compared with the normal control group, *P<0.05, **P<0.01; compared with the model control group, # P<0.05, ## P<0.01

[0088] From the above results, we can know that:

[0089] During the modeling and drug administration period, the body weight of rats in the normal control group increased steadily.

[0090] Model control group: Compared with the normal control group, the body weight of female rats increased significantly on the 8th to 25th day and the 39th day of modeling (P < 0.05 or 0.01), while there was no significant difference in the body weight of male rats.

[0091] Low-dose group of the test substance: compared with the normal control group, the body weight of male rats decreased significantly from the 4th day of administration (P < 0.05 or 0.01); compared with the model control group, the body weight of female rats decreased significantly from the 8th to the 39th day of administration (P < 0.05 or 0.01), and no other significant differences were observed.

[0092] Medium-dose group of the test substance: Compared with the normal control group, the body weight of male rats was significantly reduced on days 8 to 39 and on day 46 of administration (P < 0.05 or 0.01); compared with the model control group, the body weight of female rats was significantly reduced on days 8 to 22 and on day 36 of administration (P < 0.05), and no other significant differences were observed.

[0093] High-dose test substance group: Compared with the normal control group, the body weight of male rats decreased significantly from the 4th day of administration (P < 0.05 or 0.01); compared with the model control group, the body weight of female rats decreased significantly on the 15th day of administration (P < 0.05 or 0.01), and no other significant differences were observed.

[0094] Summary: Compared with the normal control group, the body weight of male rats in the model control group showed a downward trend, but no statistical difference was found, and the body weight of female rats increased significantly at multiple time points (P < 0.05 or 0.01). Compared with the model control group, the body weight of male rats in the low-dose and high-dose groups of the test substance showed a downward trend, but no statistical difference was found, and the body weight of female rats decreased significantly at multiple time points (P < 0.05 or 0.01). This suggests that preventive administration of the test substance can effectively reduce the body weight of rats with high sugar and high fat intake.

[0095] (3) Effect on food intake

[0096] As shown in Table 8 and Table 9.

[0097] Table 8 Effect of Tri-Glycol on Food Intake of Male High-sugar and High-fat Rats (g / rat / day, Mean±SD)

[0098]

[0099]

[0100] Table 9 Effect of Tri-Glycol on Food Intake of Female High-sugar and High-fat Rats (g / rat / day, Mean±SD)

[0101]

[0102] The above results show that compared with the normal control group rats eating normal feed, the food intake of all rats eating high-sugar and high-fat feed showed a downward trend; compared with the model control group, the food intake of rats in the low, medium and high dose groups of the test substance also showed a downward trend. This suggests that preventive administration of the test substance can effectively reduce the food intake of rats.

[0103] (4) Effects on glucose and lipid metabolism related indicators in rats

[0104] The results are shown in Tables 10, 11, 12 and 13.

[0105] Table 10 Effects of Sanzan Glue on Glucose and Lipid Metabolism in High-sugar and High-fat Rats (n=7-10, Mean±SD) Date of measurement: 2024-04-18

[0106]

[0107] Note: Compared with the normal control group, *P<0.05, **P<0.01.

[0108] Table 11 Effects of Sanzan Glue on Glucose and Lipid Metabolism in High-sugar and High-fat Rats (n=8-10, Mean±SD) Date of measurement: 2024-04-23

[0109]

[0110] Note: Compared with the normal control group, **P<0.01; compared with the model control group, # P<0.05.

[0111] Table 12 Effects of Sanzan Glue on Glucose and Lipid Metabolism in High-sugar and High-fat Rats (n=7-9, Mean±SD) Date of measurement: 2024-04-30

[0112]

[0113]

[0114] Note: Compared with the normal control group, *P<0.05, **P<0.01; compared with the model control group, # P<0.05, ## P<0.01.

[0115] Table 13 Effects of Sanzan Glue on Glucose and Lipid Metabolism in High-sugar and High-fat Rats (n=6-10, Mean±SD) Date of measurement: 2024-05-14

[0116]

[0117] Note: Compared with the normal control group, *P<0.05, **P<0.01; compared with the model control group, # P<0.05, ## P<0.01.

[0118] Before modeling: blood glucose (GLU), total cholesterol (TCHO) and triglyceride (TG) in each test group were basically similar, with no significant differences.

[0119] At 4 weeks of modeling: compared with the normal control group, the total cholesterol (TCHO), triglycerides (TG), low-density lipoprotein (LDL-C) and blood glucose (GLU) in the model control group were significantly increased (P < 0.05 or 0.01), and the high-sugar and high-fat model was successfully established. Compared with the model control group, the total cholesterol (TCHO), triglycerides (TG) and blood glucose (GLU) in the low, medium and high dose groups of the test substance showed a downward trend, and the low-density lipoprotein (LDL-C) in the low and medium dose groups of the test substance showed a downward trend, but no statistical difference was found.

[0120] After the injection of Sanzanjiao, compared with the normal control group, the blood glucose of the model control group was significantly increased (P<0.01); compared with the model control group, the blood glucose (GLU) of the low, medium and high dose groups of the test substance were significantly reduced (P<0.05).

[0121] At the end of 6 weeks of administration: compared with the normal control group, the total cholesterol (TCHO), triglyceride (TG), low-density lipoprotein (LDL-C) and blood glucose (GLU) in the model control group were significantly increased (P < 0.05 or 0.01); compared with the model control group, the blood glucose (GLU) and high-density lipoprotein (HDL-C) in the high-dose group of the test substance were significantly decreased (P < 0.05 or 0.01).

[0122] At the end of 8 weeks of administration: compared with the normal control group, the total cholesterol (TCHO), triglyceride (TG), low-density lipoprotein (LDL-C) and blood glucose (GLU) in the model control group were significantly increased (P < 0.05 or 0.01); compared with the model control group, the blood glucose (GLU) and triglyceride (TG) in the low, medium and high dose groups of the test substance were significantly decreased (P < 0.05 or 0.01), and the high-density lipoprotein (HDL-C) in the low and high dose groups of the test substance was significantly lower than that in the model group (P < 0.05).

[0123] Summary: The model created in this experiment is a high-sugar and high-fat rat model. Compared with the normal control group, the total cholesterol (TCHO), triglycerides (TG), low-density lipoprotein (LDL-C) and blood glucose (GLU) of the model control group were significantly increased (P < 0.05 or 0.01), and the high-sugar and high-fat model was successfully prepared. Compared with the model control group, after 4 weeks of administration, the total cholesterol (TCHO), triglycerides (TG), low-density lipoprotein (LDL-C) and blood glucose (GLU) in the low- and medium-dose groups of Sanzan gum showed a downward trend, and the total cholesterol (TCHO), triglycerides (TG) and blood glucose (GLU) in the high-dose group of Sanzan gum showed a downward trend; after 1 week of continued administration, the blood glucose in the low, medium and high-dose groups was significantly reduced (P < 0.05); after 6 weeks of administration, the blood glucose (GLU) and high-density lipoprotein (HDL-C) in the high-dose group were significantly reduced (P < 0.05 or 0.01); after 8 weeks of administration, the blood glucose (GLU) and triglycerides (TG) in the low, medium and high-dose groups were significantly reduced (P < 0.05 or 0.01), and the high-density lipoprotein (HDL-C) in the low and high-dose groups of the test substance was significantly lower than that in the model control group (P < 0.05). The above results indicate that Sanzan gum has a significant effect of lowering blood glucose and triglycerides in the high-sugar and high-fat rat model, but the effect of lowering total cholesterol is not obvious.

[0124] Under the conditions of this experiment, the test substance Sanzanjiao was given to high-sugar and high-fat rats by gavage at 0.14, 0.336 and 0.672g / kg, once a day, for 8 consecutive weeks. Preventive administration can effectively reduce food intake and body weight, and can significantly reduce blood sugar and triglyceride. The effective dose (0.14g / kg) is equivalent to 6.5 times the recommended human food intake based on kilogram body weight and 1 times the recommended human food intake based on body surface area.

[0125] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0126] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. Application of Sanzan gum in the preparation of lipid-lowering and blood sugar-lowering drugs.

2. The use according to claim 1, characterized in that Trisammonia gum is used to lower blood sugar and triglyceride levels.

3. The use according to claim 1, characterized in that Tri-Gel is used to reduce food intake.

4. The use according to claim 1, characterized in that Sanzan gum is used to reduce body weight.

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