Composition with hypoglycemic activity and preparation method and application thereof

Abstract

The invention provides a composition with hypoglycemic activity and a preparation method and application thereof, and belongs to the technical field of biological medicine. According to the composition disclosed by the invention, cinnamon, turmeric and red ginseng which are homologous in medicine and food are used as core raw materials and are compounded with eggshell membrane extract; after scientific components in parts by weight, all the raw materials achieve synergistic interaction, the inhibition rate on alpha-glucosidase can be remarkably increased, hyperglycemia model rat tests prove that the composition can remarkably reduce the blood sugar concentration, zebra fish tests prove that the composition can remarkably reduce the insulin content and the glucose level, the blood sugar reducing effect is remarkable, and the composition is suitable for clinical application. The traditional Chinese medicine composition can be effectively applied to blood sugar reduction and diabetes treatment. The composition is simple in component, clear in composition, high in raw material safety and free of toxic and side effects, the risk of liver injury possibly caused by combination of multiple traditional Chinese medicines is effectively avoided, and the medication safety and patient compliance are greatly improved.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, and in particular relates to a composition with hypoglycemic activity, its preparation method and uses. Background Technology

[0002] Diabetes mellitus, a prevalent chronic metabolic disease worldwide, faces numerous limitations in current clinical treatments. Oral medications often require multiple drug combinations, increasing the risk of adverse reactions and complicating treatment. Insulin therapy suffers from unstable absorption and poor adherence, highlighting the urgent need to optimize overall efficacy and convenience. Therefore, developing safe, effective, highly bioavailable, and low-side-effect-prone naturally derived hypoglycemic products has become a research hotspot in diabetes prevention and treatment. Alpha-glucosidase, an intestinal cell membrane enzyme, primarily hydrolyzes polysaccharides; thus, controlling alpha-glucosidase activity is an effective method for treating prediabetic patients and alleviating diabetic symptoms.

[0003] Traditional Chinese medicine plays a significant role in preventing and treating various chronic complications of diabetes, and its safety and versatility have garnered considerable attention. Many food-medicine homologous ingredients and natural extracts have been proven to possess potential hypoglycemic activity. Cinnamaldehyde and cinnamic acid, active components in cinnamon, can assist in regulating blood sugar through mechanisms such as improving insulin sensitivity and inhibiting carbohydrate absorption. Clinical studies have confirmed that they can significantly reduce fasting blood glucose and glycated hemoglobin levels in diabetic patients. Curcumin in turmeric has anti-inflammatory and antioxidant effects, can improve insulin resistance, and indirectly regulate glucose metabolism balance. Red ginseng, a traditional and precious Chinese medicinal material, contains ginsenosides and other components that can promote insulin secretion and improve pancreatic β-cell function, playing an important role in regulating blood sugar. Modern research has found that eggshell membranes are rich in active components such as collagen and fibronectin. Its extracts and enzymatic hydrolysates possess multiple physiological functions such as hypoglycemia, anti-inflammation, and anti-oxidation, and have the advantages of low allergenicity and high biocompatibility, showing great potential for application in the food and pharmaceutical fields.

[0004] Although the hypoglycemic activities of cinnamon, red ginseng, turmeric, and eggshell membrane extracts have been partially verified, the effects of a single substance are limited and cannot meet the complex needs of blood glucose regulation. Furthermore, there are no reports in the current technology on the rational combination of the above raw materials to exert a synergistic hypoglycemic effect. At the same time, although the application of traditional Chinese medicine formulas in the field of hypoglycemia is relatively widespread, the existing technical solutions often have the problem of complex components, which not only leads to the ambiguity of the core mechanism of action and the difficulty of quality control, but also increases the production and processing costs and the risk of medication safety. The efficacy of traditional Chinese medicine is not a simple superposition of the number of components, and scientific and simplified combination is more likely to achieve targeted synergy. Based on the hypoglycemic mechanisms and complementary effects of each ingredient, this invention abandons complex components and only combines cinnamon, red ginseng, and turmeric with eggshell membrane extract or precisely formulated combinations. Through scientific formulation, the synergistic effect between the components is achieved, which can not only improve the hypoglycemic effect and reduce the dosage of a single component, but also take into account the safety and functionality of the medication. Furthermore, due to its simple composition and clear structure, it solves the technical defects of traditional compound formulations with complex components, and provides a brand-new natural hypoglycemic option for diabetic patients. Therefore, the development of this composition with synergistic hypoglycemic activity has important practical significance and application value. Summary of the Invention

[0005] To address the aforementioned challenges, this invention optimizes and combines a hypoglycemic composition made from several traditional Chinese medicines, including cinnamon, red ginseng, and turmeric, based on modern pharmaceutical research literature. It also incorporates eggshell membrane extract to leverage the synergistic effect of each ingredient in lowering blood sugar, thereby achieving a hypoglycemic therapeutic effect.

[0006] To address this, the present invention adopts the following technical solution: The present invention provides a composition having hypoglycemic activity, comprising the following ingredients: cinnamon, turmeric and red ginseng.

[0007] Furthermore, the composition comprises the following raw materials in parts by weight: 2-9 parts cinnamon, 0.5-5 parts red ginseng, and 0.5-3 parts turmeric.

[0008] Furthermore, the composition also includes an eggshell membrane extract.

[0009] Furthermore, the eggshell membrane extract comprises 0.5-3 parts by weight.

[0010] Further, the composition comprises the following raw materials in parts by weight: 2-9 parts cinnamon, 0.5-5 parts red ginseng, 0.5-3 parts turmeric, and 0.5-3 parts eggshell membrane extract.

[0011] The present invention also provides a pharmaceutical composition comprising the aforementioned composition.

[0012] Furthermore, the pharmaceutical composition also contains pharmaceutically acceptable excipients or carriers.

[0013] The present invention also provides the use of the composition in the preparation of products having hypoglycemic effects.

[0014] Furthermore, the products include pharmaceuticals, food, or health products.

[0015] The efficacy of the Chinese herbal medicines used in this invention: Cinnamon is the dried bark of the cinnamon tree (Cinnamomum cassia), a plant belonging to the genus Cinnamomum of the Lauraceae family. Other names include: Mu Gui, Zi Gui, Da Gui, La Gui, and Gui Pi. The *Shennong Bencao Jing* (Shennong's Classic of Materia Medica) records: "Cinnamon is pungent and warm in nature, treats various diseases, nourishes the spirit, improves complexion, benefits joints, and replenishes the middle energizer and boosts qi. Long-term use leads to spiritual enlightenment, lightness of body, and longevity." It is extremely hot in nature, pungent and sweet in taste, and enters the kidney, spleen, heart, and liver meridians. It has the effects of tonifying fire and assisting yang, dispelling cold and relieving pain, warming and unblocking the meridians, and guiding fire back to its source. It is used for kidney yang deficiency, decline of the fire of the gate of life, soreness and weakness of the waist and knees, impotence and cold uterus, cold pain in the heart and abdomen, vomiting and diarrhea due to deficiency and cold, abdominal pain due to cold hernia, dysmenorrhea, and amenorrhea. Cinnamon contains abundant cinnamaldehyde, cinnamic acid, cinnamyl alcohol, volatile oil, polysaccharides, and flavonoids, which can improve blood circulation, enhance insulin sensitivity, and inhibit inflammatory responses. It has auxiliary effects in treating diabetic peripheral neuropathy, rheumatoid arthritis, coronary heart disease, dysmenorrhea, and other conditions. It also has good effects on dizziness due to yang deficiency, diarrhea due to deficiency and cold, and pain due to cold and dampness.

[0016] Red ginseng is the dried root and rhizome of ginseng, a cultivated plant belonging to the genus Panax ginseng in the family Araliaceae. It is also known as cooked ginseng, Korean ginseng, etc. The *Compendium of Materia Medica Supplement* records: "Red ginseng treats deficiency of both yin and yang, shortness of breath, cold extremities, spontaneous sweating, and sudden collapse." It is slightly warm in nature, sweet and slightly bitter in taste, and enters the spleen, lung, and heart meridians. It has the effects of greatly replenishing vital energy, restoring pulse and consolidating the body, and benefiting qi and controlling bleeding; it is used for qi deficiency and impending collapse, cold limbs and weak pulse, qi failing to control bleeding, metrorrhagia, physical weakness and fatigue, shortness of breath, and thirst due to fluid depletion. Red ginseng is rich in ginsenosides, ginseng polysaccharides, amino acids, volatile oils, vitamins, and various trace elements. It can improve the body's immunity, fight fatigue, improve cardiovascular function, and regulate endocrine function. It has auxiliary effects in treating myasthenia gravis, chronic heart failure, anemia, neurasthenia, and postoperative recovery. It also has a good effect on sallow complexion, dizziness, palpitations, insomnia, and forgetfulness caused by deficiency of qi and blood.

[0017] Curcuma longa is the dried rhizome of Curcuma longa L. of the genus Curcuma in the Zingiberaceae family. Aliases: Baodingxiang, Huangjiang, Mao Jianghuang, Shuyao, etc. It is recorded in "Compendium of Materia Medica": "Curcuma longa is mainly used to treat abdominal masses, zhuwu, lowering qi, promoting blood circulation to remove blood stasis, dispelling wind-heat, and dissipating carbuncles and swelling. Its efficacy is stronger than that of turmeric." It is warm in nature, pungent and bitter in taste, and belongs to the spleen and liver meridians, with the effects of promoting blood circulation to remove blood stasis, activating qi and relieving pain, etc.; it is used for qi stagnation and blood stasis, chest and hypochondrium stabbing pain, angina pectoris, abdominal distension and pain, amenorrhea due to blood stasis in women, dysmenorrhea, abdominal pain due to postpartum blood stasis obstruction, and rheumatic arthralgia. Curcuma longa contains rich curcumin, demethoxycurcumin, bisdemethoxycurcumin, volatile oil and polysaccharides, etc., which can antioxidant, anti-inflammatory, lower blood lipid, regulate glucose metabolism, and has adjuvant effects on the treatment of coronary heart disease, angina pectoris, chronic hepatitis, liver cirrhosis, hyperlipidemia, diabetes and other diseases; and has good curative effects on swelling and pain caused by traumatic injury and joint pain caused by cold-damp arthralgia obstruction.

[0018] The eggshell membrane extract is a product obtained by extracting and processing the dried egg membrane inside the eggshell of the domestic chicken of the Phasianidae family. Aliases: Phoenix Eggshell Extract, Egg White Membrane Extract, etc. It is recorded in "Compendium of Materia Medica": "Phoenix Eggshell is mainly used to treat chronic cough, sore throat with aphonia, scrofula and tuberculosis, and non-healing ulcers." It is neutral in nature, sweet and light in taste, and belongs to the spleen, stomach and lung meridians, with the effects of nourishing yin and clearing the lung, astringing sores, removing nebula, promoting the production of body fluid, etc.; it is used for dry cough due to lung dryness, sore throat with aphonia, nebula in the eyes, sores in the mouth and tongue, and non-healing ulcers. The eggshell membrane extract contains rich collagen, glycosaminoglycan, elastin, keratin and various active peptides, etc., which can promote tissue repair, anti-inflammatory and antioxidant, regulate joint cartilage metabolism, and has adjuvant effects on improving joint function, relieving skin dryness, promoting wound healing, etc.; and has good conditioning effects on skin pruritus caused by diabetes and joint degenerative diseases.

[0019] From the core dimensions of the properties, channels entered, and efficacy and main treatment of traditional Chinese medicines, the traditional Chinese medicine similar to cinnamon in efficacy is aconite root: extremely hot in nature, pungent and sweet in taste, toxic; entering the heart, kidney and spleen meridians. The core efficacy is to restore yang and rescue from collapse, tonify fire and assist yang, dispel cold and relieve pain, and like cinnamon, it can warm and tonify the fire of the life gate, improve symptoms such as insufficient kidney yang and fear of cold with cold limbs, and its power of dispelling cold and relieving pain is stronger, and it is often used in severe yang deficiency syndromes. The traditional Chinese medicine similar to red ginseng in efficacy is codonopsis pilosula: neutral in nature, sweet in taste; entering the spleen and lung meridians. The efficacy is to invigorate the spleen and benefit the lung, nourish blood and promote the production of body fluid, and like red ginseng, it can supplement qi and nourish blood. The medicinal property of codonopsis pilosula is mild, and the power of supplementing qi is gentle, which is suitable for chronic qi deficiency and blood deficiency syndromes and is a commonly used substitute for red ginseng.

[0020] Compared with the prior art, the present invention has the following advantages and beneficial effects: This invention provides a composition with hypoglycemic activity. The composition comprises cinnamon, turmeric, and red ginseng, all derived from medicinal and edible sources. The combination of these ingredients in specific weight proportions synergistically enhances the inhibition rate of α-glucosidase. Furthermore, in a hyperglycemic rat model experiment, it significantly reduces blood glucose concentration. The addition of eggshell membrane extract further enhances the synergistic effect of the three ingredients. Zebrafish experiments have also demonstrated that the composition significantly reduces insulin levels and glucose levels at a concentration of 10 μg / mL, exhibiting a significant hypoglycemic effect and effectively suitable for lowering blood sugar or treating diabetes. Moreover, this hypoglycemic composition uses medicinal and edible herbs as its main ingredients, eliminating complex components and precisely combining only cinnamon, turmeric, and red ginseng, along with eggshell membrane extract. Its simple composition and clear structure result in high safety and no toxic side effects, effectively avoiding the risk of liver damage that may result from long-term combined use of multiple medicinal components, thus improving drug safety and patient compliance. Attached Figure Description

[0021] Figure 1 The glucose level (mmol / L) of zebrafish after 48 h of treatment with the sample group of Example 1 (compound 1). Figure 2 The glucose level (mmol / L) of zebrafish after 48 h of treatment of samples from Examples 2-4 (compound 2-4). Figure 3 The value of insulin (mU / gprot) in zebrafish after 48 h of treatment of samples from Examples 1-4 (compound 1-4) is given. Detailed Implementation

[0022] The present invention will be further illustrated below with reference to specific embodiments. Unless otherwise specified, the raw materials, reagents or devices used in the following embodiments can be obtained from conventional commercial sources or by existing known methods.

[0023] It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer.

[0024] Unless otherwise specified, all materials and reagents used in the following examples are commercially available. In the specific examples of this invention, cinnamon, turmeric, and red ginseng were purchased from the market by dry weight. Eggshell membrane extract was purchased from Xi'an Xihai Biotechnology Co., Ltd., product number XH503 (1 kg).

[0025] Data analysis was performed using GraphPad Prism 8.0 statistical software. Quantitative data are expressed as mean ± standard error (mean ± SEM). Independent samples t-tests were used for comparisons between two groups, and one-way ANOVA was used for comparisons among multiple groups. p < 0.05 was considered statistically significant.

[0026] Examples and Comparative Examples Examples and comparative examples of formulations of the compositions of the present invention having hypoglycemic activity are shown in Table 1 below: Table 1. Formulation composition of examples and comparative examples raw material Example 1 Example 2 Example 3 Example 4 Example 5 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Comparative Example 6 Comparative Example 7 Cinnamon 5.5 2 9 5.5 5.5 10 5.5 5.5 <![CDATA[5.5 # ]]> 1 10 Red Ginseng 2.75 5 0.5 2.75 2.75 5.5 <![CDATA[2.75 * ]]> 2.75 6 0.5 turmeric 1.75 0.5 3 1.75 1.75 4.5 4.5 1.75 1.75 3 0.5 Eggshell membrane extract 0.5 3 0.5 (Note: In Table 1, the "*" mark in Comparative Example 4 indicates that the red ginseng in the formula was replaced with an equal amount of Codonopsis pilosula, and the "#" mark in Comparative Example 5 indicates that the cinnamon in the formula was replaced with an equal amount of Aconitum carmichaelii.) The preparation methods of the traditional Chinese medicine compositions in Examples 1-5 and Comparative Examples 1-7 are as follows: Weigh out the full amount of cinnamon, red ginseng, turmeric, and eggshell membrane extract (if any) according to Table 1, add 10 times the amount of water, soak for 30 minutes, then heat and reflux to extract twice, 2 hours each time. Filter and combine the two filtrates, and concentrate under reduced pressure at 80℃ to an extract with a relative density of 1.23-1.26 (50℃) to obtain the hypoglycemic composition.

[0027] Test Example 1: Activity Test of Hypoglycemic Composition on α-Glucosidase Inhibition Screening for α-glucosidase inhibitor activity can be performed by detecting the in vitro enzymatic reaction between the enzyme and its substrate, p-nitrophenyl-β-D-galactopyranoside (pNPG, a maltose analog). After the substrate is added to the enzyme, it is catalyzed and broken down into p-nitrophenol (PNP) and glucose. PNP is a colored substance with maximum absorption at approximately 405 nm; the inhibitory activity of the sample can be calculated by measuring the OD value.

[0028] Take 500 μL of phosphate buffer solution (pH 6.8, 0.067 mol / L), 500 μL of α-glucosidase (0.1 U / mL), and 100 μL of the test sample solution (10 μg / mL, hypoglycemic compositions prepared in Examples 1-5 and Comparative Examples 1-7) and mix them thoroughly. Then, after incubating at 37 °C for 20 min, add 500 μL of pNPG (4.0 mmol / L), mix thoroughly, and continue to react at 37 °C for 30 min. After the reaction is completed, add methanol to terminate the reaction. Calculate the α-glucosidase inhibition rate. The experimental results are shown in Table 2. α-glucosidase inhibition rate (%) = [(C a -C b +C c ) / Ca ×100%; Where, C a is the concentration of pNPG after the buffer solution reacts with the enzyme solution and the substrate; C b is the concentration of pNPG after the sample reacts with the enzyme solution and the substrate; C c is the concentration of pNPG after the sample reacts with the buffer solution and the substrate.

[0029] Table 2 Inhibition rate of the hypoglycemic composition of the present invention on α-glucosidase (n = 3) Sample to be tested α-glucosidase inhibition rate (%) Example 1 Group 84.3±5.6 Example 2 group 81.5±4.7 Example 3 Group 82.8±5.2 Example 4 group 89.2±6.4 Example 5 group 92.4±5.5 Comparative Example 1 42.5±4.1 Comparative Example 2 36.1±3.8 Comparative Example 3 Groups 47.3±5.7 Comparative Example 4 Groups 65.2±4.8 Comparative Example 5 Groups 50.7±6.2 Comparative Example 6 Groups 67.5±5.1 Comparative Example 7 Groups 52.4±4.7 The compositions of the examples and comparative examples of the present invention can all inhibit the activity of α-glucosidase, but there are differences in the inhibition rate on α-glucosidase. Among them, the inhibition rates of the hypoglycemic compositions of Examples 1-5 on α-glucosidase are all greater than 80%, which is significantly higher than the inhibition effect of the comparative examples. From the results of Example 1 and Comparative Examples 1-3, it can be seen that there is an obvious synergistic effect among cinnamon, red ginseng and turmeric. The inhibition effect of the composition lacking any one component on the enzyme is significantly reduced. Moreover, from the results of replacing red ginseng with codonopsis pilosula and cinnamon with aconite, it can be seen that the raw material composition of the present invention is not randomly combined, and its selection has prominent substantive features and remarkable progress. In addition, eggshell membrane extracts were added to Examples 4-5 respectively, and their effects of inhibiting enzyme activity are also higher than those of Examples 1-3, indicating that the eggshell membrane extract also has a certain enhancement effect on the inhibition of enzyme activity.

[0030] Test Example 2 Therapeutic effect of the hypoglycemic composition on hyperglycemic rats Test sample: Hypoglycemic compositions prepared from Examples 1-5 and Comparative Examples 1-7.

[0031] Animal grouping: SPF-grade SD rats, half male and half female, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., certificate number SCXK(Beijing)2012-0001; after the animals were purchased, the feeding environment temperature was (25 ±1)°C, with natural light, fed with normal feed and allowed to eat freely, and adapted to the environment for 7 days. They were randomly divided into a normal control group, a model control group, a positive control group, Example 1 to 5 groups, and Comparative Example 1 to 7 groups, with 8 rats in each group, a total of 120 rats.

[0032] Construction of type 2 diabetes (T2DM) rat model: Except for 8 rats in the normal control group, the other 112 rats were fed with a high-sugar diet for 4 weeks, then fasted for 12 hours without water deprivation, and intraperitoneally injected with streptozotocin (STZ) at a dose of 45 mg / kg at one time. When injecting, it was dissolved with a citrate buffer solution (pH 4.2-4.5) at a concentration of 1%. 72 hours later, blood glucose levels of rats in each group were detected by tail vein blood sampling at the same time for 3 consecutive days. If all 3 detections were greater than 16 mmol / L, it was considered that the T2DM model was successfully established.

[0033] Administration method: The example groups and comparative groups were administered the hypoglycemic compositions prepared in Examples 1-5 and Comparative Examples 1-7 by gavage once daily for 2 weeks, at a dose of 0.5 g / kg, with a volume of 0.1 mL / 10 g. The blank control group and the model control group were administered the same volume of distilled water once daily for 2 weeks. The positive control group was administered 150 mg / (kg·d) metformin hydrochloride tablets (Harbin Laibotong Pharmaceutical Co., Ltd., batch number: National Drug Approval Number H23021803) by gavage. During the administration period, the rats in the blank group were fed a normal diet, while the rats in the other groups were fed a high-sugar diet.

[0034] Measurement of indicators: After two weeks of drug administration, rats in each group were fasted for 12 hours but allowed free access to water. Blood was then collected from their tails to measure blood glucose levels. The results are shown in Table 3.

[0035] Table 3. Hypoglycemic effect of the hypoglycemic composition on hyperglycemic rats (n=8) Sample to be tested Before administration (mmol / L) Two weeks after administration (mmol / L) normal control group 7.41±1.35 7.26±1.28 Model control group 23.58±3.51** <![CDATA[23.41±1.92 ** ]]> Positive control group 22.72±3.24** <![CDATA[7.40±1.43 ### ]]> Example 1 Group 22.43±3.14** <![CDATA[10.73±1.01 ## ]]> Example 2 group 23.64±3.07** <![CDATA[11.24±1.16 ## ]]> Example 3 Group 23.03±2.43** <![CDATA[10.56±1.24 ## ]]> Example 4 group 22.76±2.76** <![CDATA[9.18±1.18 ## ]]> Example 5 group 22.61±1.88** <![CDATA[7.27±0.86 ### ]]> Comparative Example 1 22.45±2.54** 16.48±1.24 Comparative Example 2 21.95±2.23** 17.25±1.34 Comparative Example 3 Groups 22.45±3.12** 15.87±1.25 Comparative Example 4 Groups 23.12±2.97** 18.43±1.31 Comparative Example 5 Groups 22.76±3.42** 16.25±1.53 Comparative Example 6 Groups 22.55±2.65** 17.21±1.95 Comparative Example 7 Groups 23.18±3.30** 16.94±1.58 (Note: Compared with the control group, ** indicates P < 0.01, * indicates P < 0.05; compared with the model control group, ## indicates P < 0.001.) ## # indicates P < 0.01, # indicates P < 0.05. As shown in Table 3, the blood glucose level of the model rats before drug administration was 23.58±3.51mmol / L (greater than 16mmol / L), which was significantly higher than that of the blank group, and there was a significant statistical difference between the two, indicating that the model was successfully established. The other positive control group, the example group and the comparative group were also successfully modeled.

[0036] Two weeks after administration, the positive control group, Examples 1-5, and Comparative Examples 1-7 all reduced the blood glucose concentration in rats. Among them, the positive control group and Examples 1-5 showed statistically significant differences in blood glucose levels compared with the model control group (p < 0.01), and the positive control group and Example 5 showed a more significant hypoglycemic effect than Comparative Examples 1-7 (p < 0.001). This indicates that the cinnamon, red ginseng, and turmeric of the present invention can synergistically increase the hypoglycemic effect of the composition. In addition, the hypoglycemic effect is even better with the addition of eggshell membrane extract, indicating that the hypoglycemic composition of the present invention has good application prospects in the fields of medicine and health products.

[0037] Experiment Example 3: Evaluation of the Adjunctive Hypoglycemic Efficacy of Zebrafish Model Animal rearing and treatment: Adult zebrafish were housed in a recirculating aquaculture system at a temperature maintained at 28°C, with a photocycle of 14 hours (h) light / 10 hours (h) darkness. They were fed three times daily. To obtain embryos, sexually mature zebrafish were paired at a 1:1 female-to-male ratio at night and separated by a baffle. The baffle was removed within one hour of the start of the next photocycle to allow for natural spawning. The collected embryos were placed in a 1×E3 solution (10 cm diameter petri dish) containing 0.3 ppm methylene blue and cultured in a 28.5°C artificial climate chamber (14 h / 10 h light / dark) until the experimental treatment stage.

[0038] (1) Assisted in lowering blood sugar I Experimental groups: normal control group, model control group, positive control group (metformin 400 μg / mL), and sample compound group 1 (i.e. Example 1 group, with 3 concentration gradients: 2.5 μg / mL, 5 μg / mL, and 10 μg / mL).

[0039] Methods: Zebrafish 5 days post-fertilization (5 dpf) were randomly selected and exposed to samples in 6-well plates (3 mL / well), with 30 fish per well. The normal control group was cultured normally, while the model control group, positive control group, and sample group were alternately induced to develop a high glucose model by being treated with 0.1% egg yolk powder (7 h daytime treatment) and 3% glucose (17 h overnight treatment). After 48 h of exposure, 3 zebrafish were collected in 0.6 mL centrifuge tubes, the culture medium was discarded, and 25 μL of anhydrous ethanol was added to completely immerse the zebrafish. The tubes were dried at 65 °C for 60 min, and 3 μL of ultrapure water was added to each tube. After shaking and centrifugation, the tubes were allowed to stand for 10 min, and glucose levels were measured using a glucometer to evaluate the auxiliary hypoglycemic effect of the samples.

[0040]

[0041] (2) Assisted in lowering blood sugar II Experimental groups: normal control group, model control group, positive control group (metformin 400 μg / mL), sample compound 2 (Example 2 group, 10 μg / mL), sample compound 3 (Example 3 group, 10 μg / mL), and sample compound 4 (Example 4 group, 10 μg / mL).

[0042] The processing method is the same as above.

[0043] (3) Insulin content Experimental groups: normal control group, model control group, positive control group (metformin 400 μg / mL), sample compound 1 (Example 1 group, 10 μg / mL), sample compound 2 (Example 2 group, 10 μg / mL), sample compound 3 (Example 3 group, 10 μg / mL), and sample compound 4 (Example 4 group, 10 μg / mL).

[0044] Methods: Zebrafish 5 days post-fertilization (5 dpf) were randomly selected and exposed to samples in 6-well plates (3 mL / well), with 30 fish per well. The normal control group was cultured normally, while the model control group, positive control group, and sample group were alternately induced to develop a high glucose model by being treated with 0.1% egg yolk powder (daytime treatment for 7 h) and 3% glucose (overnight treatment for 17 h). After 48 h of exposure, samples were collected, and insulin levels were measured using an insulin detection kit.

[0045] Experimental results: (1) Assisted in lowering blood sugar I The results are detailed in Table 4. Figure 1 After 48 h of treatment, compared with the model control group, the glucose level of zebrafish in Example 1 group (Compound 1 group) was significantly reduced, revealing that Compound 1 has an auxiliary hypoglycemic effect. According to the calculation formula for auxiliary hypoglycemic effect, the auxiliary hypoglycemic effect of Compound 1 at the test concentrations of 2.5, 5, and 10 μg / mL was 62%, 70%, and 80%, respectively.

[0046] Table 4 Results of the auxiliary hypoglycemic experiment of Compound Sample 1 (Example 1) group (n = 10)

[0047] (Note: Compared with the model control group, *** represents p < 0.001.) (2) Assisted in lowering blood sugar II The results are detailed in Table 5. Figure 2 After 48 hours of treatment, compared with the model control group, the glucose levels of zebrafish in the Example 2-4 groups (compound 2-4 groups) were significantly reduced, revealing that compound 2, 3, and 4 have an auxiliary hypoglycemic effect. According to the calculation formula for the auxiliary hypoglycemic effect, the auxiliary hypoglycemic effects of compound 2, 3, and 4 were 67%, 52%, and 73%, respectively.

[0048] Table 5. Results of the auxiliary hypoglycemic experiment of compound preparation 2-4 (Examples 2-4) group (n = 10) Group Concentration (μg / mL) Glucose level (mmol / L) normal control group - <![CDATA[0.630 ± 0.0153 *** ]]> Model control group - 2.30 ± 0.0471 Metformin 400 <![CDATA[0.970 ± 0.0423 *** ]]> Compound 2 (Example 2) group 10 <![CDATA[1.18 ± 0.0327 *** ]]> Compound 3 (Example 3) group 10 <![CDATA[1.43 ± 0.0473 *** ]]> Compound 4 (Example 4) group 10 <![CDATA[1.08 ± 0.0291 *** ]]> (Note: Compared with the model control group, *** represents p < 0.001.) (3) Insulin content The results are detailed in Table 6. Figure 3 After 48 hours of treatment, compared with the model control group, the insulin content of zebrafish in the Example 1-4 groups (compound 1-4 groups) was significantly reduced.

[0049] Table 6. Experimental results of insulin content in zebrafish after treatment with compound preparations 1-4 (Examples 1-4) (n = 3) Group Concentration (μg / mL) Insulin levels (mU / gprot) normal control group - <![CDATA[0.217 ± 0.000760 *** ]]> Model control group - 0.247 ± 0.000719 Metformin 400 <![CDATA[0.233 ± 0.00247 ** <!-- 7 -->]]> Compound 1 (Example 1) group 10 <![CDATA[0.236 ± 0.00132 ** ]]> Compound 2 (Example 2) group 10 <![CDATA[0.236 ± 0.00195 ** ]]> Compound 3 (Example 3) group 10 <![CDATA[0.237 ± 0.00323 * ]]> Compound 4 (Example 4) group 10 <![CDATA[0.234 ± 0.00249 ** ]]> (Note: Compared with the model control group, * represents p < 0.05, ** represents p < 0.01, and *** represents p < 0.001.) In the zebrafish model evaluation experiment on the efficacy of auxiliary hypoglycemic agents, the hypoglycemic compositions of Examples 1-4 of the present invention all showed significant auxiliary hypoglycemic effects, specifically by reducing glucose levels and insulin content in zebrafish.

[0050] The above description, in conjunction with specific preferred embodiments, provides a further detailed explanation of the present invention. It should not be construed that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, various simple deductions or substitutions can be made without departing from the concept of the present invention, and all such modifications and substitutions should be considered within the scope of protection of the present invention.

Claims

1. A composition having hypoglycemic activity, characterized in that, It is composed of the following ingredients: cinnamon, turmeric, and red ginseng.

2. The composition according to claim 1, characterized in that, It consists of the following ingredients in parts by weight: 2-9 parts cinnamon, 0.5-5 parts red ginseng, and 0.5-3 parts turmeric.

3. The composition according to any one of claims 1-2, characterized in that, The composition also includes an eggshell membrane extract.

4. The composition according to claim 3, characterized in that, The eggshell membrane extract is present in 0.5-3 parts by weight.

5. The composition according to any one of claims 3-4, characterized in that, The composition comprises the following raw materials in parts by weight: 2-9 parts cinnamon, 0.5-5 parts red ginseng, 0.5-3 parts turmeric, and 0.5-3 parts eggshell membrane extract.

6. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the composition according to any one of claims 1-5.

7. The pharmaceutical composition according to claim 6, characterized in that, The pharmaceutical composition also contains pharmaceutically acceptable excipients or carriers.

8. The use of the composition according to any one of claims 1-5 in the preparation of a product having hypoglycemic effect.

9. The application according to claim 8, characterized in that, The products include pharmaceuticals, food, or health supplements.

Images