Use of dendrobium extract in preparation of medicine for treating type 2 diabetes

Abstract

The application discloses application of a dendrobium extract in preparation of a medicine for treating type 2 diabetes, and mainly relates to the technical field of dendrobium. Specifically, the application provides a medicine for treating type 2 diabetes and containing the dendrobium extract, improves stability of components in the medicine for treating type 2 diabetes in an acid, neutral or alkaline environment, and slows down blood sugar increase. Compared with the prior art, the medicine for treating type 2 diabetes prepared by the application takes the dendrobium extract as a core, is matched with a synergistic emulsion, a rhizoma polygonati extract, a fructus corni extract, a salvia miltiorrhiza extract, a momordica charantia extract, vitamin B and trace elements of chromium, and through cooperation with traditional Chinese medicine components, can synergistically regulate blood sugar metabolism, enhance insulin sensitivity and improve microcirculation, can stabilize blood sugar, improve insulin resistance and lipid metabolism, improve immunity and cardiovascular function, and provides a safe and effective medicine for prevention and treatment of type 2 diabetes.

Description

Technical Field

[0001] This invention relates to the field of Dendrobium technology, and more particularly to the application of a Dendrobium extract in the preparation of a drug for treating type 2 diabetes. Background Technology

[0002] Type 2 diabetes mellitus (T2DM) is a chronic disease characterized by hyperglycemia, with its core mechanisms being abnormal insulin secretion and insulin resistance (i.e., relative insulin deficiency). In Traditional Chinese Medicine (TCM) theory, diabetes falls under the category of "Xiao Ke" (wasting and thirsting syndrome), with pathogenesis involving excessive stomach fire, lung dryness damaging body fluids, and kidney deficiency failing to retain water. TCM often uses neutral, cold, and warm herbs to tonify Qi and nourish Yin, clear heat and generate fluids, and promote blood circulation to relieve symptoms. Furthermore, improving gut microbiota is crucial for T2DM patients, as its metabolic products can influence the condition through regulation of metabolism, immunity, and other aspects. Gut microbiota imbalance caused by factors such as a high-fat diet can exacerbate disease progression.

[0003] Dendrobium officinale, an orchid species, is recorded in the Chinese Pharmacopoeia as having the effects of nourishing the stomach and yin, clearing heat and promoting body fluids, which aligns with the medicinal properties used in traditional Chinese medicine to treat "diabetes." It contains various active ingredients such as alkaloids, flavonoids, and polysaccharides. Its water extract can stabilize blood sugar by improving insulin sensitivity, promoting glycogen synthesis, and reducing reactive oxygen species and inflammatory factors, thus protecting tissues and organs from oxidative stress and chronic inflammatory damage caused by hyperglycemia. Besides single-dose administration, Dendrobium officinale compound preparations are also commonly used to treat type 2 diabetes mellitus (T2DM). For example, when combined with Astragalus membranaceus, Rehmannia glutinosa, and Salvia miltiorrhiza, it can significantly reduce blood glucose and lipid levels in T2DM mice and improve liver pathological structure; its combined use with metformin and other Western medicines can also enhance efficacy. For T2DM patients, improving the gut microbiota is also particularly important. Dendrobium officinale compound preparations can regulate the gut microbiota, improve the abundance of beneficial bacteria in the mouse gut, promote the production of short-chain fatty acids, and protect the intestinal mucosa, thereby mitigating the adverse effects of gut microbiota imbalance on T2DM patients and further assisting in alleviating the condition.

[0004] CN118161571A discloses a hypoglycemic compound preparation, its preparation method, and its uses. The compound preparation includes extracts of Dendrobium officinale, Astragalus membranaceus, mulberry leaf, kudzu root, and American ginseng. This invention extracts water-soluble components from these traditional Chinese medicines, then mixes and blends these extracted water-soluble components in different proportions to prepare the compound preparation. It achieves synergistic, complementary, and enhanced efficacy, showing significant effects in treating or alleviating diabetes and lowering blood sugar. It can be applied to foods, beverages, snack foods, and health products with auxiliary hypoglycemic functions. However, this method does not address the component stability of the water-soluble extracts in type 2 diabetes medication applications, nor does it study their long-term effects on stabilizing blood sugar.

[0005] CN109820959A discloses the application of Dendrobium officinale extract in the preparation of drugs for the prevention and treatment of dyslipidemia. The extract is prepared from Dendrobium officinale or its water-extracted residue through drying, pulverization, reflux extraction, separation and purification, and further drying. The total flavonoid content is 2.5-80%, containing flavonoid C-glycosides with apigenin as the aglycone. Its application can comprehensively improve various metabolic disorders, such as lowering blood pressure and blood sugar, regulating blood lipids, improving microcirculation and blood viscosity, etc., and can be used in drugs and health foods for metabolic diseases, showing broad development prospects. However, this specification does not mention the optimization of component stability in the application of this extract in type 2 diabetes drugs, nor does it address the issue of improving blood glucose stability. Summary of the Invention

[0006] In view of the shortcomings of existing technologies, such as insufficient component stability and unsatisfactory long-term blood sugar lowering effects, the purpose of this invention is to provide the application of Dendrobium extract in the treatment of type 2 diabetes. This drug can not only improve the stability of active ingredients in acidic, neutral, and alkaline environments, but also slow down the rise in blood sugar, thereby improving metabolic disorders in patients with type 2 diabetes.

[0007] To achieve the above objectives, this invention proposes the application of Dendrobium officinale extract in the preparation of drugs for treating type 2 diabetes. Specifically, the aqueous extract of Dendrobium officinale is scientifically formulated with synergistic emulsions, Polygonatum sibiricum extract, Cornus officinalis extract, Salvia miltiorrhiza extract, Momordica charantia extract, B vitamins, and trace elements such as chromium. The resulting composition can achieve synergistic effects through multiple targets and pathways: on the one hand, it improves insulin sensitivity and promotes glucose metabolism; on the other hand, it enhances the stability and bioavailability of the components in vivo, thereby achieving the effects of stabilizing blood sugar and improving diabetic complications.

[0008] More specifically, the preparation method of the drug for treating type 2 diabetes is as follows:

[0009] The following ingredients are mixed: Dendrobium extract, synergistic emulsion, Polygonatum extract, Cornus officinalis extract, Salvia miltiorrhiza extract, bitter melon extract, B complex vitamins, and trace elements such as chromium. The mixture is then dried and ground through a 40-60 mesh sieve to obtain the final product.

[0010] The preparation method of the synergistic emulsion is as follows:

[0011] Citrus pectin and vitamin C are added to water and stirred for 0.5-1 hour to obtain the aqueous phase. Lycopene is added to medium-chain triglycerides and stirred for 0.5-1 hour to obtain the oil phase. The aqueous phase and oil phase are mixed and stirred for 0.5-1 hour. Geraniol is added, homogenized for 4-6 minutes, and ultrasonically dispersed for 5-15 minutes to obtain the synergistic emulsion, which is stored at 0-5℃.

[0012] The amounts of the Dendrobium extract, synergistic emulsion, Polygonatum extract, Cornus officinalis extract, Salvia miltiorrhiza extract, bitter melon extract, B complex vitamins, and chromium trace element are 150-250 parts, 45-55 parts, 30-50 parts, 20-40 parts, 20-40 parts, 10-20 parts, 2-4 parts, and 2-4 parts, respectively; where parts refer to parts by weight.

[0013] The mass ratio of the citrus pectin, vitamin C, water, lycopene, medium-chain triglycerides, and geraniol is 6-10:0.4-0.6:350-450:0.5-1.5:45-55:0.4-0.6.

[0014] The B complex vitamins are a combination of vitamins B1, B2, B3, B5, B6, B7, B9, and B12.

[0015] The chromium trace element is chromium-rich yeast.

[0016] The extraction method of the Dendrobium extract is as follows:

[0017] Add Dendrobium officinale powder to water, decoct for 40-50 minutes, and filter to obtain the first decoction. Add the residue to water, decoct for 20-40 minutes, and filter to obtain the second decoction. Combine the two decoctions and concentrate to obtain Dendrobium officinale extract, which is stored at 4℃.

[0018] The mass ratio of Dendrobium officinale powder to water during the first decoction is 1:45-55.

[0019] The mass ratio of filter residue to water during the second decoction is 1:15-25.

[0020] The drug solutions are combined and concentrated to 45-55 wt% of their initial mass.

[0021] Dendrobium extract is rich in active ingredients such as polysaccharides, which can nourish yin and moisten dryness, benefit the stomach and promote the production of body fluids. It is a core component for improving type 2 diabetes, and can enhance the body's immunity and regulate blood sugar metabolism.

[0022] Polygonatum extract contains polysaccharides and other components, which can replenish qi and nourish yin, strengthen the spleen and moisten the lungs. It works synergistically with Dendrobium extract to enhance the body's sensitivity to insulin and stabilize blood sugar levels.

[0023] Cornus officinalis extract has the effects of tonifying the liver and kidneys, astringing essence and stopping sweating. It can improve the body's metabolism and regulate blood sugar. When combined with Dendrobium officinale extract, it enhances the effect of the combination on improving diabetes.

[0024] Salvia miltiorrhiza extract contains components such as tanshinone, which can promote blood circulation, remove blood stasis, and improve microcirculation. It helps prevent and improve diabetic complications. In combination with Dendrobium officinale extract, it has a positive effect on the cardiovascular system of diabetic patients.

[0025] Bitter melon extract contains components such as shizurin, which have the effect of lowering blood sugar and regulating the body's sugar metabolism. When combined with dendrobium extract, it helps control blood sugar and improve metabolic disorders in diabetic patients.

[0026] B vitamins can help regulate glucose metabolism, and when combined with the above ingredients, they can further enhance the body's ability to regulate blood sugar. Chromium, a trace element, can enhance the effect of insulin and, in synergy with ingredients such as Dendrobium extract, helps stabilize blood sugar levels.

[0027] The beneficial effects of this invention are:

[0028] Compared with existing technologies, this invention uses Dendrobium officinale extract as the core ingredient, combined with Polygonatum sibiricum extract, Cornus officinalis extract, Salvia miltiorrhiza extract, bitter melon extract, B vitamins, and trace elements such as chromium. These ingredients exhibit synergistic effects without antagonism, and all comply with the requirements of the "Administrative Measures for the Catalogue of Raw Materials and Health Functions of Health Foods." The drug for treating type 2 diabetes in this invention can significantly stabilize indicators such as fasting blood glucose and glycated hemoglobin, improve insulin sensitivity, and reduce insulin resistance. It can also effectively regulate blood glucose levels in patients with type 2 diabetes, regulate lipid metabolism, enhance immunity, and improve cardiovascular function. Furthermore, no significant toxic side effects have been observed with long-term use, demonstrating good application prospects and market potential. This invention provides a safe and effective drug for the prevention and treatment of type 2 diabetes. Attached Figure Description

[0029] Figure 1 Venn diagram showing the common target of Dendrobium officinale extract and type 2 diabetes;

[0030] Figure 2 Network diagram of Dendrobium extract components and targets;

[0031] Figure 3 List of core ingredients of Dendrobium extract;

[0032] Figure 4 A PPT network diagram of Dendrobium extract;

[0033] Figure 5 The results are KEGG and GO enrichment analyses, where A is the KEGG enrichment analysis result and B is the GO enrichment analysis result.

[0034] Figure 6Insulin tolerance test (ITT) was performed on mice in the normal group, model group, and Example 4 drug treatment group, and the area under the curve was recorded. Group G represents the normal group, D represents the Example 4 drug treatment group, and M represents the model group.

[0035] Figure 7 The insulin content in the serum of each mouse was determined by enzyme-linked immunosorbent assay (ELISA), where group G represents the normal group, group D represents the drug administration group in Example 4, and group M represents the model group.

[0036] Figure 8 The values ​​represent the T-SOD content in mouse pancreatic tissue, where group G represents the normal group, group D represents the drug administration group in Example 4, and group M represents the model group. Detailed Implementation

[0037] The parameters of the specific chemical substances used in the examples are from the following sources:

[0038] The extraction method of bitter melon extract is as follows: Fresh bitter melon is dried, crushed, sieved, and defatted with petroleum ether. The treated bitter melon is extracted twice with 70wt% ethanol at a material-to-liquid ratio of 1:14 g / mL, an extraction temperature of 60℃, and an extraction time of 180 minutes. The extracts are combined, centrifuged, and the supernatant is concentrated and dried to obtain bitter melon extract.

[0039] The extraction method of kudzu root extract is as follows: Select dried kudzu root (originating from Yunnan), remove impurities, wash it clean and crush it into 100-mesh coarse kudzu root powder; extract the coarse kudzu root powder twice with 95wt% ethanol, with a material-to-liquid ratio of 1:10 g / mL, an extraction temperature of 60℃ and an extraction time of 180 minutes, combine the extracts, centrifuge and concentrate and dry the supernatant to obtain kudzu root extract.

[0040] The extraction method of Ophiopogon japonicus extract is as follows: Ophiopogon japonicus (produced in Sichuan) is ground through a 40-mesh sieve to obtain coarse powder; the coarse powder is extracted once with 80wt% ethanol, the material-to-liquid ratio is 1:14 g / mL, the extraction temperature is 80℃, and the extraction time is 360 minutes. After centrifugation, the supernatant is concentrated and dried to obtain Ophiopogon japonicus extract.

[0041] Polygonatum extract: Polygonatum (produced in Hunan) was sliced ​​and soaked in 95% ethanol at room temperature. The extract was collected every 24 hours, concentrated under reduced pressure, and circulated 6 times. The extracts were combined, centrifuged, and the supernatant was concentrated and dried to obtain Polygonatum extract.

[0042] Cornus officinalis extract: Add dried Cornus officinalis fruit (from Shanxi) to 6 times the amount of water and decoct for 2 hours. Repeat 3 times, combine the 3 decoctions, filter and concentrate the filtrate to obtain Cornus officinalis extract.

[0043] Danshen extract: Add 10 times the amount of water to Danshen (produced in Henan), sonicate for 10 minutes, decoct for 20 minutes, repeat the decoction 3 times, combine the 3 decoctions, filter and concentrate the filtrate at 60℃ to obtain Danshen extract.

[0044] The B complex vitamins are a combination of various vitamins B1, B2, B3, B5, B6, B7, B9, and B12.

[0045] Chromium-enriched yeast: Manufacturer: Shanghai Nuoshen Food Trading Co., Ltd., Model: Cr2000.

[0046] Example 1

[0047] The extraction method of Dendrobium officinale extract is as follows: 80mg of Dendrobium officinale powder is added to 4000mg of water, decocted for 45 minutes and filtered to obtain the first decoction; the residue is added to 1500mg of water, decocted for 30 minutes and filtered to obtain the second decoction; the two decoctions are combined and concentrated to 2800mg to obtain Dendrobium officinale extract, which is stored at 4℃.

[0048] Example 2

[0049] A method for preparing a drug for treating type 2 diabetes is as follows:

[0050] 2000 mg of Dendrobium officinale extract prepared in Example 1, 400 mg of Polygonatum sibiricum extract, 300 mg of Cornus officinalis extract, 300 mg of Salvia miltiorrhiza extract, 6 mg of vitamin B1, 6 mg of vitamin B2, 6 mg of vitamin B6, 6 mg of vitamin B9, 6 mg of vitamin B12, and 30 mg of chromium-enriched yeast were mixed, dried, and ground through a 50-mesh sieve to obtain the drug for treating type 2 diabetes.

[0051] Example 3

[0052] A method for preparing a drug for treating type 2 diabetes is as follows:

[0053] 2000 mg of Dendrobium officinale extract prepared in Example 1, 400 mg of Polygonatum sibiricum extract, 300 mg of Cornus officinalis extract, 300 mg of Salvia miltiorrhiza extract, 150 mg of Momordica charantia extract, 6 mg of vitamin B1, 6 mg of vitamin B2, 6 mg of vitamin B6, 6 mg of vitamin B9, 6 mg of vitamin B12, and 30 mg of chromium-enriched yeast were mixed, dried, and ground through a 50-mesh sieve to obtain the drug for treating type 2 diabetes.

[0054] Comparative Example 1

[0055] A method for preparing a drug for treating type 2 diabetes is as follows:

[0056] 2000 mg of Dendrobium officinale extract prepared in Example 1, 400 mg of Polygonatum sibiricum extract, 300 mg of Cornus officinalis extract, 300 mg of Salvia miltiorrhiza extract, 150 mg of Pueraria lobata extract, 6 mg of vitamin B1, 6 mg of vitamin B2, 6 mg of vitamin B6, 6 mg of vitamin B9, 6 mg of vitamin B12, and 30 mg of chromium-enriched yeast were mixed, dried, and ground through a 50-mesh sieve to obtain the drug for treating type 2 diabetes.

[0057] Comparative Example 2

[0058] A method for preparing a drug for treating type 2 diabetes is as follows:

[0059] 2000 mg of Dendrobium officinale extract prepared in Example 1, 400 mg of Polygonatum sibiricum extract, 300 mg of Cornus officinalis extract, 300 mg of Salvia miltiorrhiza extract, 150 mg of Ophiopogon japonicus extract, 6 mg of vitamin B1, 6 mg of vitamin B2, 6 mg of vitamin B6, 6 mg of vitamin B9, 6 mg of vitamin B12, and 30 mg of chromium-enriched yeast were mixed, dried, and ground through a 50-mesh sieve to obtain the drug for treating type 2 diabetes.

[0060] Example 4

[0061] A method for preparing a drug for treating type 2 diabetes is as follows:

[0062] 2000 mg of Dendrobium officinale extract prepared in Example 1, 500 mg of synergistic emulsion, 400 mg of Polygonatum sibiricum extract, 300 mg of Cornus officinalis extract, 300 mg of Salvia miltiorrhiza extract, 150 mg of Momordica charantia extract, 6 mg of vitamin B1, 6 mg of vitamin B2, 6 mg of vitamin B6, 6 mg of vitamin B9, 6 mg of vitamin B12, and 30 mg of chromium-enriched yeast were mixed, dried, and ground through a 50-mesh sieve to obtain the drug for treating type 2 diabetes.

[0063] The preparation method of the synergistic emulsion is as follows:

[0064] 8 mg of citrus pectin and 0.5 mg of vitamin C were added to 400 mg of water and stirred for 0.5 hours to obtain the aqueous phase. 1 mg of lycopene was added to 50 mg of medium-chain triglycerides and stirred for 0.5 hours to obtain the oil phase. The aqueous phase and oil phase were mixed and stirred for 0.5 hours. 0.5 mg of geraniol was added, and the mixture was homogenized at 10,000 r / min for 5 minutes and ultrasonically dispersed for 10 minutes to obtain the synergistic emulsion, which was stored at 4°C.

[0065] Example 5

[0066] A method for preparing a drug for treating type 2 diabetes is as follows:

[0067] 2000 mg of Dendrobium officinale extract prepared in Example 1, 500 mg of synergistic emulsion, 400 mg of Polygonatum sibiricum extract, 300 mg of Cornus officinalis extract, 300 mg of Salvia miltiorrhiza extract, 150 mg of Momordica charantia extract, 6 mg of vitamin B1, 6 mg of vitamin B2, 6 mg of vitamin B6, 6 mg of vitamin B9, 6 mg of vitamin B12, and 30 mg of chromium-enriched yeast were mixed, dried, and ground through a 50-mesh sieve to obtain the drug for treating type 2 diabetes.

[0068] The preparation method of the synergistic emulsion is as follows:

[0069] 8 mg of citrus pectin and 0.5 mg of vitamin C were added to 400 mg of water and stirred for 0.5 hours to obtain the aqueous phase. 1 mg of lycopene was added to 50 mg of medium-chain triglycerides and stirred for 0.5 hours to obtain the oil phase. The aqueous phase and oil phase were mixed and stirred for 0.5 hours, homogenized at 10000 r / min for 5 minutes, and ultrasonically dispersed for 10 minutes to obtain the synergistic emulsion, which was stored at 4℃.

[0070] Example 6

[0071] A method for preparing a drug for treating type 2 diabetes is as follows:

[0072] 2000 mg of Dendrobium officinale extract prepared in Example 1, 500 mg of synergistic emulsion, 400 mg of Polygonatum sibiricum extract, 300 mg of Cornus officinalis extract, 300 mg of Salvia miltiorrhiza extract, 150 mg of Momordica charantia extract, 6 mg of vitamin B1, 6 mg of vitamin B2, 6 mg of vitamin B6, 6 mg of vitamin B9, 6 mg of vitamin B12, and 30 mg of chromium-enriched yeast were mixed, dried, and ground through a 50-mesh sieve to obtain the drug for treating type 2 diabetes.

[0073] The preparation method of the synergistic emulsion is as follows:

[0074] 8 mg of citrus pectin and 0.5 mg of vitamin C were added to 400 mg of water and stirred for 0.5 hours to obtain the aqueous phase. 50 mg of medium-chain triglycerides were added and stirred for 0.5 hours to obtain the oil phase. The aqueous phase and oil phase were mixed and stirred for 0.5 hours. 0.5 mg of geraniol was added, and the mixture was homogenized at 10,000 r / min for 5 minutes and ultrasonically dispersed for 10 minutes to obtain the synergistic emulsion, which was stored at 4°C.

[0075] Example 7

[0076] A method for preparing a drug for treating type 2 diabetes is as follows:

[0077] 2000 mg of Dendrobium officinale extract prepared in Example 1, 400 mg of Polygonatum sibiricum extract, 300 mg of Cornus officinalis extract, 300 mg of Salvia miltiorrhiza extract, 150 mg of Momordica charantia extract, 6 mg of vitamin B1, 6 mg of vitamin B2, 6 mg of vitamin B6, 6 mg of vitamin B9, 6 mg of vitamin B12, 30 mg of chromium-enriched yeast, 8 mg of citrus pectin, 1 mg of lycopene, and 0.5 mg of geraniol were mixed, dried, ground, and sieved to obtain the drug for treating type 2 diabetes.

[0078] Test Example 1

[0079] The Dendrobium extract obtained by the extraction method in Example 1 was used to search for T2DM-related targets in the Genecards and OMIM databases, and duplicates were removed to form a disease target set. The intersection of the component target set and the disease target set was taken to obtain the intersection of potential targets. The Venn diagram was drawn using the Venny2.1 website to obtain the Venn diagram of common targets of Dendrobium extract and type 2 diabetes. Figure 1 ).

[0080] Potential targets of Dendrobium officinale extract were imported into Cytoscape 3.9.1 software to draw a network diagram of Dendrobium officinale extract-components-targets. Figure 2 The relationship between components and targets was then analyzed and visualized. Using degree as the screening criterion, the top 10 components were selected as core components, including 3-O-methylgallic acid, dentin, and naringenin, resulting in the core component table of Dendrobium extract. Figure 3 ).

[0081] Potential targets were imported into the STRING database, and a PPI network was established. Network topology analysis and visualization were performed using Cytoscape 3.9.1 software. The top 12 targets were selected as core targets based on their degree value. Higher degree values ​​indicate a darker color, signifying a closer relationship and greater importance with other targets. The top 12 targets by degree value were identified as AKT1, TNF, ALB, PPARG, SRC, EGFR, STAT3, ESR1, HIF1A, NFKB1, BCL2, and CASP3, resulting in a PPT network diagram of Dendrobium officinale extract. Figure 4 ).

[0082] KEGG and GO enrichment analyses were performed on Dendrobium extract. In the KEGG enrichment analysis graph, the smaller the P.Value, the darker the color, the more target sites involved, and the larger the bubbles, the better the KEGG enrichment analysis results. Figure 5-A) shows that 166 potential pathways were identified through co-enrichment, and the 14 pathways with the lowest P.Value were plotted as a bubble chart. The results indicate that *Dendrobium officinale* may act on type 2 diabetes mellitus (T2DM) through pathways such as the PI3K-Akt signaling pathway, the MAPK signaling pathway, and the insulin resistance signaling pathway. GO enrichment analysis results ( Figure 5 -B) shows that the GO analysis yielded 841 results, including 579 biological processes (BP), 189 cellular components (CC), and 73 molecular functions (MF). The 10 results with the lowest P.Values ​​were selected for analysis and visualization. BP involves biological functions such as cell signaling, chromatin remodeling, and inflammatory responses; MF includes the cell membrane, cytoplasm, and nucleus; and CC includes functions related to protein binding, ATP binding, and zinc ion binding.

[0083] Conclusion: Analysis of the Dendrobium extract obtained by the extraction method in Example 1 identified 62 active ingredients and 210 potential targets. Drug-component-target network topology analysis revealed 10 core components, including 3-O-methylgallic acid, dentin, and naringenin. PPI analysis identified 12 core targets, mainly including TNF, AKT1, and ALB. GO enrichment analysis showed that Dendrobium exerts its effects through biological functions such as cell signal transduction, chromatin remodeling, and inflammatory responses; cellular components such as the cell membrane, cytoplasm, and nucleus; and molecular functions including protein binding, ATP binding, and zinc ion binding. KEGG pathway enrichment analysis indicated that the Dendrobium extract obtained by the extraction method in Example 1 can improve type 2 diabetes mellitus (T2DM) through cancer pathways, insulin resistance, and insulin signaling pathways.

[0084] Test Example 2

[0085] Animal experiment results

[0086] Fifty male C57 / BL6 mice, weighing 18-22g, were randomly divided into a normal group, a model group, and groups receiving treatments in Examples 2-7 and Comparative Examples 1-2. The mice were housed in a constant temperature environment of 20-22℃ and humidity of 40%-60%, using a 12-hour light cycle followed by a 12-hour dark cycle, with free access to sterilized feed and water. After one week of acclimatization to a standard diet, the model group, groups receiving treatments in Examples 2-7 and Comparative Examples 1-2 were given a high-fat diet, while the normal group received a standard diet. This acclimatization period continued for 6 weeks.

[0087] In the sixth week, mice in the model group, Examples 2-7, and Comparative Examples 1-2 were fasted for 12 hours and then injected intraperitoneally with 100 mg / kg STZ solution. Mice in the normal group were injected intraperitoneally with an equal volume of 1 wt% citrate buffer. On days 3 and 5 after injection, mice were fasted for 12 hours and then blood was collected by tail clipping. After removing the first drop of blood, blood glucose was measured using a blood glucose meter. Mice with fasting blood glucose values ​​>11.1 mmol / ml on two separate occasions were considered to be T2DM mice.

[0088] The drugs for treating type 2 diabetes prepared by the methods of Examples 2-7 and Comparative Examples 1-2 were used as test samples. Mice in the treatment group were given 25 g / kg of 10 wt% of the test aqueous solution of Examples 2-7 and Comparative Examples 1-2 by gavage every day. The model group and the blank group were given an equal volume of distilled water by gavage. Fasting blood glucose of mice in the normal group, model group, and treatment groups of Examples 2-7 and Comparative Examples 1-2 were recorded on days 7, 14, 21 and 28, respectively. Blood was collected by tail clipping. After the first drop of blood was removed, blood glucose was measured using a blood glucose meter. The blood glucose changes are summarized in Table 1.

[0089] After blood glucose measurement on day 28, insulin tolerance test (ITT) was performed on mice in the normal group, model group, and the drug administration group of Example 4, and the area under the curve (AUC) was recorded. Figure 6 Mice were euthanized after blood was collected by enucleation of the eyeballs, and pancreatic tissue and cecum were harvested for testing. The insulin content in the serum of each mouse was determined by enzyme-linked immunosorbent assay (ELISA). Figure 7 The hydroxylamine method was used to determine the T-SOD content in mouse pancreatic tissue. Figure 8 ), Figure 6-8 Group G represents the normal group, Group D represents the treatment group of Example 4, and Group M represents the model group. Fresh feces were collected from the cecum for 16S rDNA high-throughput sequencing and bioinformatics analysis. The diversity of intestinal flora and dominant bacterial genera of mice in the normal group, model group, and treatment group of Example 3 were compared, as shown in Table 2.

[0090] Conclusion: From Figure 6-8 The results showed that, compared with the normal group, the area under the curve (AUC) of the ITT test in the model control group mice was significantly increased, serum insulin levels were significantly decreased, and pancreatic T-SOD levels were significantly reduced, demonstrating that T2DM mice were in a state of insulin resistance, reduced insulin secretion, and decreased pancreatic antioxidant capacity. Meanwhile, compared with the model control group, the area under the curve (AUC) of the ITT test in the drug-treated group mice was significantly decreased, serum insulin levels were significantly increased, and pancreatic T-SOD levels were significantly increased.

[0091] Table 1

[0092]

[0093] Note: The difference between Example 4 and Example 3 was statistically significant (P<0.05); the difference between Example 4 and Examples 5-7 was statistically significant (P<0.05).

[0094] Table 1 shows that at week 0, compared with the normal group, the fasting blood glucose levels of mice in the model group and the drug-treated group were higher. On days 7 and 14, compared with the model group, the increase in fasting blood glucose levels in the drug-treated group was slower. Although the mice in the drug-treated group remained in a hyperglycemic state, the trend was slower compared to the model group. On days 21 and 28 of the intervention, compared with the model group, the increase in fasting blood glucose levels in the drug-treated group was slower, showing a significant difference, indicating that the drug for treating type 2 diabetes of the present invention has a hypoglycemic effect.

[0095] The drug formulation for treating type 2 diabetes in Example 2 is a combination of Dendrobium officinale extract, Polygonatum sibiricum extract, Cornus officinalis extract, Salvia miltiorrhiza extract, B vitamins, and trace element chromium. Examples 3 and Comparative Examples 1-2, based on Example 2, respectively added Bitter Melon extract, Kudzu root extract, and Ophiopogon japonicus extract. The bitter melon extract and Dendrobium officinale extract, among other components, may enhance the hypoglycemic effect through synergistic effects. The bitter melon extract contains momordicin, which can mimic insulin action and promote glucose uptake. This, combined with the insulin-sensitizing effect of Dendrobium officinale polysaccharide and the glucose absorption-inhibiting effect of Polygonatum sibiricum polysaccharide, forms a multi-target synergistic effect, thereby enhancing hypoglycemic efficiency. Kudzu root extract contains puerarin, which mainly works by improving insulin sensitivity. Ophiopogon japonicus extract contains ophiopogonin, which focuses on regulating liver glycogen metabolism. Compared to bitter melon extract, the synergistic effect of kudzu root extract and Ophiopogon japonicus extract is weaker.

[0096] Example 4 is based on Example 3, with the addition of a synergistic emulsion containing lycopene and geraniol. The lycopene and geraniol are encapsulated using citrus pectin as an emulsifier, which significantly improves the stability and bioavailability of lycopene and geraniol. Lycopene can scavenge free radicals and protect pancreatic β cells, while geraniol can inhibit α-glucosidase activity. Together with the insulin mimicry effect of bitter melon extract and the metabolic regulation effect of Dendrobium officinale, they form a closed-loop synergy of "antioxidant-hypoglycemic-metabolic regulation", further enhancing the overall hypoglycemic effect. The synergistic emulsion in Example 5 did not contain geraniol, thus lacking its inhibitory effect on α-glucosidase and its antioxidant synergy with lycopene. It could only assist in lowering blood sugar through the single component of lycopene, failing to form a multi-target synergistic effect, therefore its effect was weaker than that of Example 3. The synergistic emulsion in Example 6 also lacked lycopene, thus lacking lycopene's ability to scavenge free radicals and protect pancreatic β-cells. It relied solely on geraniol to inhibit α-glucosidase, reducing the number of synergistic targets and failing to form a synergistic blood sugar lowering effect with geraniol. Example 7 added citrus pectin, lycopene, and geraniol alone. The components did not form a stable emulsion structure, making them easily destroyed or difficult to absorb, thus failing to exert a synergistic effect, and its blood sugar lowering effect was lower than that of Example 3.

[0097] Table 2

[0098]

[0099] Note: The difference between the drug-treated group and the normal group in Example 4 was statistically significant (P<0.05); the difference between the drug-treated group and the model group in Example 4 was statistically significant (P<0.05).

[0100] Compared with the normal group, there were no significant differences in serum insulin levels and pancreatic T-SOD levels in the treatment group; compared with the model group, the treatment group had higher serum insulin levels and pancreatic T-SOD levels, and a smaller area under the curve in the ITT test. The results of the gut microbiota α-diversity study showed that the gut microbiota diversity of the T2DM model group mice was reduced compared with the normal group; compared with the model group, the diversity was increased in the Dendrobium officinale treatment group. Furthermore, after intervention with Dendrobium officinale aqueous extract, the abundance of bacteria such as Olsenella and Desulfovibrio in the treatment group mice was increased, while the abundance of bacteria such as Dubosiella, Enterococcus, and Romboutsia was decreased.

[0101] Test Example 3

[0102] Take 0.05g of the drug for treating type 2 diabetes prepared by the methods of Examples 2-7 and Comparative Examples 1-2, respectively, and dilute it in 5g of water to use as test samples. Each example and comparative example is further divided into three test groups: acidic, neutral, and alkaline, with a total of 5 test samples in each group.

[0103] pH adjusters were added to the acidic, neutral, and alkaline test groups respectively, adjusting the pH of the solution in the acidic test group to 3.0, the pH of the solution in the neutral test group to 7.0, and the pH of the solution in the alkaline test group to 11.0. The pH adjusters were 2 mol / L NaOH and 5 mol / L HCl. After standing for 30 minutes, the absorbance at λ=228 nm was measured using distilled water as a reference. The results are summarized in Table 3 to determine the stability of the Dendrobium extract in the drugs for treating type 2 diabetes obtained by the preparation methods of Examples 2-7 and Comparative Examples 1-2 under different pH conditions.

[0104] Table 3

[0105]

[0106] Note: The difference between Example 4 and Example 3 was statistically significant (P<0.05); the difference between Example 4 and Examples 5-7 was statistically significant (P<0.05).

[0107] In Test Example 3, the drugs for treating type 2 diabetes prepared by the methods in Examples 2-7 and Comparative Examples 1-2 were placed under acidic (pH=3.0), neutral (pH=7.0), and alkaline (pH=11.0) conditions to measure the absorbance of Dendrobium officinale extract. Examples 2-3 and Comparative Examples 1-2 relied solely on the compatibility of the extracts themselves to reduce component loss. Examples 3 and Comparative Examples 1-2, based on Example 2, added bitter melon extract, Ophiopogon japonicus extract, and Pueraria lobata extract, respectively. The organic acids in bitter melon extract helped stabilize Dendrobium officinale extract, and the saponins in Ophiopogon japonicus extract formed hydrogen bonds with Dendrobium officinale polysaccharides to aid in stabilization. The stabilizing effect of Pueraria lobata extract on Dendrobium officinale extract was relatively weak. Example 4 added a synergistic emulsion containing citrus pectin and geraniol. Citrus pectin formed a dense oil-in-water structure, reducing the adsorption loss of Dendrobium officinale extract from other components, and geraniol inhibited local oxidation. Therefore, it had the highest Dendrobium officinale extract content and the best stability. The synergistic emulsion of Example 5 did not contain geraniol, thus lacking antioxidant protection, and localized oxidation resulted in a lower content than in Example 4. The synergistic emulsion of Example 6 did not contain lycopene, lacking the synergistic effect of lycopene, and its stability was lower than that of Example 4. Compared to Example 4, Example 7 involved the direct mixing of citrus pectin and geraniol, resulting only in physical adsorption and a lower protective effect on the Dendrobium extract.

[0108] The preferred embodiments of the present invention have been described in detail above. It should be understood that those skilled in the art can make numerous modifications and variations based on the concept of the present invention without creative effort. Therefore, all technical solutions that can be obtained by those skilled in the art based on the concept of the present invention through logical analysis, reasoning, or limited experimentation on the basis of existing technology should be within the scope of protection defined by the claims.

Claims

1. A method for the preparation of a medicament for the treatment of type 2 diabetes, characterized in that, It comprises the following steps: mixing Dendrobium extract, synergistic emulsion, Rhizoma Polygonati extract, Fructus Corni extract, Salvia miltiorrhiza extract, Momordica charantia extract, compound vitamin B group, and trace element chromium, drying, grinding through 40-60 mesh screen, and obtaining; The use amount of the Dendrobium extract, synergistic emulsion, Rhizoma Polygonati extract, Fructus Corni extract, Salvia miltiorrhiza extract, Momordica charantia extract, compound vitamin B group, and trace element chromium is respectively 150-250 parts, 45-55 parts, 30-50 parts, 20-40 parts, 20-40 parts, 10-20 parts, 2-4 parts, and 2-4 parts, wherein, the part refers to mass part. The preparation method of the synergistic emulsion is as follows: The citrus pectin and vitamin C are added into water and stirred for 0.5-1 hour to obtain an aqueous phase, lycopene is added into medium-chain triglyceride and stirred for 0.5-1 hour to obtain an oil phase, the aqueous phase and the oil phase are mixed and stirred for 0.5-1 hour, myrciin is added, homogenized for 4-6 minutes, and ultrasonic dispersed for 5-15 minutes to obtain the synergistic emulsion, which is stored at 0-5℃. The use amount of the citrus pectin, vitamin C, water, lycopene, medium-chain triglyceride, and myrciin is in a mass ratio of 6-10:0.4-0.6:350-450:0.5-1.5:45-55:0.4-0.

6. The extraction method of the Dendrobium extract is as follows: iron skin Dendrobium powder is added into water, decocted for 40-50 minutes, and then filtered to obtain first decocting medicinal liquid; the filter residue is added into water, decocted for 20-40 minutes, and then filtered to obtain second decocting medicinal liquid; the two medicinal liquids are combined and concentrated to obtain the Dendrobium extract. The extraction method of the Momordica charantia extract is as follows: fresh Momordica charantia is dried, crushed, sieved, and then degreased by petroleum ether, the treated Momordica charantia is extracted by 70wt% ethanol for 2 times, the solid-liquid ratio is 1:14 g / mL, the extraction temperature is 60℃, and the extraction time is 180 minutes, the extraction liquid is combined, centrifuged, and then the supernatant is concentrated and dried to obtain the Momordica charantia extract. The extraction method of the Rhizoma Polygonati extract is as follows: Rhizoma Polygonati is sliced, and then soaked in 95% ethanol at room temperature, the extraction liquid is collected every 24 hours, concentrated under reduced pressure, and the cycle is repeated for 6 times, the extraction liquid is combined, centrifuged, and then the supernatant is concentrated and dried to obtain the Rhizoma Polygonati extract. The extraction method of the Fructus Corni extract is as follows: Fructus Corni is added into 6 times water for decoction, the decoction time is 2 hours, and the decoction is repeated for 3 times, the decocting liquid of the 3 times is combined, filtered, and then the filtrate is concentrated to obtain the Fructus Corni extract. The extraction method of the Salvia miltiorrhiza extract is as follows: Salvia miltiorrhiza is added into 10 times water, ultrasonic dispersed for 10 minutes, decocted for 20 minutes, and the decoction is repeated for 3 times, the decocting liquid of the 3 times is combined, filtered, and then the filtrate is concentrated at 60℃ to obtain the Salvia miltiorrhiza extract.

2. The production method according to claim 1, characterized by: In the extraction method of the Dendrobium extract, the mass ratio of iron skin Dendrobium powder to water in the first decoction is 1:45-55.

3. The production method according to claim 1, wherein In the extraction method of the Dendrobium extract, the mass ratio of the filter residue to water in the second decoction is 1:15-25.

4. The production method according to claim 1, wherein In the extraction method of the Dendrobium extract, the medicinal liquid is combined and concentrated to 45-55wt% of the initial mass.

5. The production method according to claim 1, wherein The complex vitamin B group is a combination of vitamin B1, vitamin B2, vitamin B3, vitamin B5, vitamin B6, vitamin B7, vitamin B9 and vitamin B12.

6. The production method according to claim 1, wherein The trace element of chromium element is chromium-rich yeast.

7. A drug for treating type 2 diabetes, characterized by comprising the compound of claim 1. The preparation method is as claimed in any one of claims 1-6.

8. The use of the drug as claimed in claim 7 for preparing a drug for treating type 2 diabetes.

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