Liquid-increasing freeze-drying composition and application thereof

Abstract

The invention provides a liquid-increasing freeze-drying composition and application thereof, and relates to the technical field of Chinese patent medicines. The liquid-increasing freeze-drying composition is obtained by performing vacuum freeze-drying on a liquid-increasing stock solution, the stock solution of the fluid increasing decoction consists of an extracting solution obtained by water extraction of liriope spicata and an extracting solution obtained by water extraction and alcohol precipitation of radix scrophulariae / rehmannia. The liquid-increasing freeze-drying composition can be used as a component of a medicine or a medicine composition for treating diabetes mellitus.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicines and relates to a liquid-enhanced freeze-dried composition and an application thereof. Background Art

[0002] Zengye Decoction, composed of 30g Scrophularia ningpoensis, 24g Ophiopogon japonicus, and 24g Rehmannia glutinosa, with Scrophularia ningpoensis as the main ingredient, simultaneously nourishes yin, softens hard masses, and purges heat. Zengye Decoction was first described in the Qing Dynasty febrile disease expert Wu Jutong's treatise on febrile diseases, "Treatise on Warm Diseases," to treat constipation caused by fluid depletion due to Yangming febrile diseases, a syndrome characterized by "no water, no boat." Wu Jutong proposed the core concept of this prescription as "increasing water to move the boat," addressing constipation caused by intestinal dryness by nourishing yin and moistening dryness rather than simply purging. Its creation stemmed from the limitations of the Chengqi Decoction-like formulas in the Treatise on Febrile Diseases, emphasizing adaptive adjustments for patients with yin deficiency.

[0003] Diabetes mellitus is a group of metabolic diseases characterized by chronic hyperglycemia. Its core mechanism is impaired insulin secretion (absolute or relative deficiency) and / or impaired insulin action (insulin resistance), leading to disrupted sugar, fat, and protein metabolism, which can lead to multi-organ dysfunction and failure over the long term. Clinical studies have found that modified Zengye Decoction, when used in combination with conventional Western medicine, has a significant effect on lowering blood sugar and lipids. It can improve insulin sensitivity, improve insulin resistance, and inhibit inflammation, effectively alleviating clinical symptoms and improving patients' quality of life. Modern research suggests that Zengye Decoction may also exert its glucose-lowering effects and improve insulin resistance by improving the "fat-islet axis," regulating the intestinal flora, and ameliorating chronic inflammation. Furthermore, it may also be involved in pathways such as amino acid metabolism, energy metabolism, and glucose metabolism.

[0004] Traditional liquid-enhancing decoctions are liquid preparations that present challenges such as poor stability, inconvenient transportation, and rapid deterioration during transportation and storage. To meet long-term storage requirements, the addition of antibacterial agents to inhibit microbial growth is often necessary, but this can pose safety risks and increase formulation complexity. Summary of the Invention

[0005] In order to solve the above technical problems, the present invention provides a liquid-enhanced freeze-dried composition and its application.

[0006] The technical solutions of the present invention are as follows:

[0007] A liquid-enhancing freeze-dried composition is obtained by vacuum freeze-drying a liquid-enhancing soup stock solution.

[0008] Preferably, the fluid-increasing soup stock solution is composed of Ophiopogon japonicus extract and Scrophularia ningpoensis / Rehmannia glutinosa extract.

[0009] More preferably, the Radix Ophiopogonis extract is obtained by extracting Radix Ophiopogonis from Radix Ophiopogonis with water.

[0010] More preferably, the relative density of the Ophiopogon japonicus extract at 60° C. is 1.15-1.18.

[0011] More preferably, the Scrophulariaceae / Rehmannia glutinosa extract is obtained by extracting Scrophulariaceae and Rehmannia glutinosa with water and precipitating with alcohol.

[0012] More preferably, the relative density of the Scrophulariaceae / Rehmannia glutinosa extract at 60° C. is 1.17-1.20.

[0013] More preferably, the water extraction is performed by decocting for 1-3 times, and the decocting time for each time is 0.5-2 hours.

[0014] Preferably, the vacuum freeze-drying comprises the steps of freezing, vacuum sublimation and drying;

[0015] The sublimation temperature is no more than 0°C.

[0016] More preferably, the drying temperature is 30-55°C.

[0017] A use of the enriched lyophilized composition according to any of the above embodiments as a component of a drug or pharmaceutical composition for treating diabetes and / or promoting the recovery of pancreatic islet cell function.

[0018] The beneficial effects of the present invention are as follows: the present invention converts the fluid-enhancing soup into a fluid-enhancing freeze-dried composition after vacuum freeze-drying, which is more convenient to transport and store, has more stable product quality, and has a better effect on diabetes than the fluid-enhancing soup stock solution. DETAILED DESCRIPTION

[0019] The technical solution of the present invention is further illustrated and described below through specific implementation methods.

[0020] In order to improve the quality stability and effect of the fluid-increasing soup, on the one hand, the present invention provides a fluid-increasing freeze-dried composition obtained by vacuum freeze-drying the fluid-increasing soup stock solution.

[0021] The present invention converts the fluid-enhancing decoction stock solution into a freeze-dried composition of the fluid-enhancing decoction through vacuum freeze-drying. The freeze-dried composition of the fluid-enhancing decoction is in powder form at room temperature. Compared to fluid-enhancing decoction, it has better quality stability, is more convenient to transport, and has a longer shelf life without the addition of antibacterial agents or preservatives. Furthermore, the freeze-dried powder obtained through vacuum freeze-drying has more stable quality and is more easily dissolved and dispersed in water during use. The freeze-dried composition of the fluid-enhancing decoction obtained by the present invention can be stored in vials.

[0022] The above-mentioned vacuum freeze-drying includes freezing, vacuum sublimation and drying steps, that is, the liquid-enhancing soup stock solution is frozen at a temperature not higher than -20°C (such as -45°C, -40°C, -35°C, etc.) to form a solid, and then evacuated to a vacuum degree not exceeding 30Pa. Most of the water in the liquid-enhancing soup frozen into a solid is sublimated and removed, and then the temperature is raised to 30-55°C for drying to further remove the water more completely.

[0023] For the above freezing, the specific operation method is not particularly limited. For example, the temperature is preset to a freezing temperature (such as -45°C, -40°C, -35°C, etc.), and the temperature is lowered at 0.2-0.5°C / min, and maintained for 2-4 hours. For the above sublimation, the specific operation is not particularly limited. For example, the vacuum system is started, the vacuum degree of the drying chamber is quickly reduced to 10-30 Pa, and the temperature is increased to the sublimation temperature at a rate of 0.5-1°C / h. The sublimation temperature may not exceed 0°C (such as -20 to -10°C), and maintained for 12-48 hours. For the above drying, the specific operation is not particularly limited. For example, after the sublimation is completed, the vacuum degree is adjusted to 10-20 Pa, the temperature is increased to 30-55°C at a rate of 0.5-1°C / min, and maintained for 2-8 hours. Then, the vacuum degree is adjusted to 40-50 Pa, and the temperature is lowered to 15-25°C at a rate of 0.5-1°C / min, and maintained for 0.5-3 hours.

[0024] The present invention has found that the sublimation temperature of the vacuum freeze-drying process not exceeding 0° C. is more conducive to obtaining a liquid-enriched freeze-dried composition with stable quality.

[0025] In some embodiments, the fluid-enhancing decoction concentrate comprises an extract of Radix Ophiopogonis and an extract of Scrophulariae / Rehmannia glutinosa. This extract allows the effective ingredients from Radix Ophiopogonis, Scrophulariae, and Rehmannia glutinosa to be extracted, maintaining the efficacy of the fluid-enhancing decoction. The Scrophulariae / Rehmannia glutinosa extract refers to an extract of a blend of Scrophulariae and Rehmannia glutinosa.

[0026] In some embodiments, the extract of Radix Ophiopogonis is obtained by water extraction of Radix Ophiopogonis. Radix Ophiopogonis extract can be directly obtained by water extraction of Radix Ophiopogonis. For water extraction, Radix Ophiopogonis can be decocted in water, the number of decoctions can be 1-3 times, and the time of each decoction can be 0.5-2 hours, and the decoction is then concentrated to a certain concentration or density. For example, the method of water extraction of Radix Ophiopogonis can be as follows: at room temperature, Radix Ophiopogonis is soaked in water for 0.5-1 hour, the weight ratio of Radix Ophiopogonis to water is 1:8-1:20, and decocted for 1 hour, repeated twice, the decoctions are combined, filtered and concentrated, and the concentration method can be heating and negative pressure, such as a temperature of 60-70°C and a pressure of -0.08 to -0.04MPa.

[0027] In some embodiments, the relative density of the Liriope japonicus extract at 60° C. is 1.15-1.18. In the present invention, relative density refers to the density relative to pure water at 4° C. The relative density of the Liriope japonicus extract at 60° C. can be 1.15, 1.16, 1.17, 1.18, etc., preferably 1.16.

[0028] In some embodiments, the Scrophulariaceae / Rehmannia glutinosa extract is obtained by water extraction and alcohol precipitation of Scrophulariaceae and Rehmannia glutinosa. The extraction of Scrophulariaceae and Rehmannia glutinosa requires water extraction and alcohol precipitation, wherein alcohol precipitation involves precipitating the aqueous extract with an alcoholic solvent (e.g., ethanol) and removing the sediment to extract the active ingredients from Scrophulariaceae and Rehmannia glutinosa, effectively removing impurities and improving the purity and quality stability of the extract. For example, the preparation method of the Scrophulariaceae / Rehmannia glutinosa extract can be as follows: Scrophulariaceae and Rehmannia glutinosa are soaked in water for 0.5-1 hour, the weight ratio of the Scrophulariaceae / Rehmannia glutinosa mixture to water is 1:50-1:30, and the mixture is decocted twice, each decocting time is 1 hour, the decoctions are combined, filtered, and concentrated under reduced pressure at 60-70°C to obtain a concentrate, the relative density of the concentrate at 60°C can be 1.15-1.20; after the concentrate is cooled to room temperature, ethanol is added (the amount of ethanol added is such that the alcohol content in the concentrate is 50-65%, the alcohol content is measured using an alcohol meter), the concentrate is allowed to stand, and then centrifuged. The filtrate is collected, and after removing the ethanol from the filtrate, water is added to adjust the relative density at 60°C to 1.17-1.20, which is the Scrophulariaceae / Rehmannia glutinosa extract. For example, the relative density of the Scrophulariaceae / Rehmannia glutinosa extract at 60°C can be 1.17, 1.17, 1.18, 1.20, etc., preferably 1.18.

[0029] In another aspect, the present invention further provides a use of the rehydrated lyophilized composition described in any of the above embodiments as a drug or component of a pharmaceutical composition for treating diabetes and / or promoting pancreatic islet cell function recovery. The rehydrated lyophilized composition of the present invention has a good effect of lowering blood sugar and promoting pancreatic islet cell function recovery. It can be used alone as a drug for treating diabetes and promoting pancreatic islet cell function recovery, or it can be combined with other drugs to form a pharmaceutical composition for treating diabetes and / or promoting pancreatic islet cell function recovery.

[0030] The technical solution of the present invention is further described and illustrated below based on various embodiments.

[0031] Example 1

[0032] Take 30g of Scrophularia, 24g of Ophiopogon japonicus, and 24g of Rehmannia glutinosa.

[0033] Add 240g of water to soak Radix Ophiopogonis for 0.5h, decoct for 1h, repeat twice, combine the two decoctions, filter, and evaporate the filtrate under reduced pressure at 60℃ to a relative density of 1.16 (60℃) to obtain Radix Ophiopogonis extract.

[0034] Scrophulariaceae and Rehmannia root were mixed and soaked in 540 g of water for 0.5 h. The mixture was decocted for 1 h, repeated twice, and the decoctions were combined and filtered. The filtrate was concentrated under reduced pressure at 60° C. to a relative density of 1.18 (60° C.) to obtain a concentrate. The concentrate was cooled to room temperature, ethanol was added to a concentration of 60% alcohol, and the mixture was allowed to stand at room temperature for 24 h. The mixture was centrifuged at 3000 rpm for 15 min, and the filtrate was collected. The filtrate was evaporated at 70° C. to recover the ethanol, and water was added to adjust the relative density to 1.18 (60° C.) to obtain the Scrophulariaceae / Rehmannia root extract.

[0035] The above-mentioned Radix Ophiopogonis extract and Scrophulariae / Rehmanniae extract are combined and mixed evenly to obtain a liquid-enhancing soup stock solution. The liquid-enhancing soup stock solution is placed in a fully automatic vacuum freeze dryer and subjected to pre-freezing, sublimation, and drying steps to prepare a powdered liquid-enhancing freeze-dried composition.

[0036] The vacuum freeze-drying process of this embodiment is shown in Table 1 below.

[0037] Table 1

[0038] stage Temperature (℃) Cooling / heating rate (℃ / min) Holding time (h) Vacuum degree (Pa) Pre-freeze -45 0.5 3 / Take time to sublimate -10 0.7 35 20 dry 40 1 6 20

[0039] Example 2

[0040] The difference between this embodiment and embodiment 1 is that in embodiment 1, the vacuum freeze-drying process is adjusted as shown in Table 2. The remaining steps remain unchanged.

[0041] Table 2

[0042] stage Temperature (℃) Cooling / heating rate (℃ / min) Holding time (h) Vacuum degree (Pa) Pre-freeze -45 0.5 3 / Take time to sublime -5 0.7 35 20 dry 40 1 6 20

[0043] Example 3

[0044] The difference between this embodiment and embodiment 1 is that in embodiment 1, the vacuum freeze-drying process is adjusted as shown in Table 3. The remaining steps remain unchanged.

[0045] Table 3

[0046] stage Temperature (℃) Cooling / heating rate (℃ / min) Holding time (h) Vacuum degree (Pa) Pre-freeze -45 0.5 3 / Take time to sublime 0 0.7 35 20 dry 40 1 6 20

[0047] Example 4

[0048] The difference between this embodiment and embodiment 1 is that in embodiment 1, the vacuum freeze-drying process is adjusted as shown in Table 4. The remaining steps remain unchanged.

[0049] Table 4

[0050] stage Temperature (℃) Cooling / heating rate (℃ / min) Holding time (h) Vacuum degree (Pa) Pre-freeze -45 0.5 3 / Take time to sublimate 5℃ 0.7 35 20 dry 40 1 6 20

[0051] Example 5

[0052] The difference between this embodiment and embodiment 3 is that in embodiment 3, the vacuum freeze-drying process is adjusted as shown in Table 5. The remaining steps remain unchanged.

[0053] Table 5

[0054] stage Temperature (℃) Cooling / heating rate (℃ / min) Holding time (h) Vacuum degree (Pa) Pre-freeze -45 0.5 3 / Take time to sublimate 0 0.7 20 20 dry 40 1 6 20

[0055] The appearance and moisture content of the rehydrated freeze-dried compositions obtained in Examples 1-5 are shown in Table 6 below, where the moisture content was tested using the Karl Fischer method.

[0056] Table 6

[0057] serial number Product Appearance Moisture content (%) Example 1 good 3.5 Example 2 good 3.0 Example 3 good 3.2 Example 4 A little shrinking 4.0 Example 5 good 1.5

[0058] Therefore, the results in Table 6 show that a vacuum sublimation temperature not exceeding 0° C. is beneficial for obtaining a liquid-enriched freeze-dried composition with a better appearance.

[0059] The following Examples 6-10 are process adjustments based on Example 5.

[0060] Example 6

[0061] The difference between this embodiment and embodiment 5 is that the Scrophulariaceae / Rehmannia glutinosa extract is prepared without alcohol precipitation, that is, the Scrophulariaceae / Rehmannia glutinosa extract is the concentrated solution in embodiment 5. The remaining steps remain unchanged.

[0062] Example 7

[0063] The difference between this embodiment and embodiment 5 is that the amount of ethanol added is such that the alcohol content is adjusted from 60% to 40%. The other steps remain unchanged.

[0064] Example 8

[0065] The difference between this embodiment and embodiment 5 is that the amount of ethanol added is such that the alcohol content is adjusted from 60% to 50%. The other steps remain unchanged.

[0066] Example 9

[0067] The difference between this embodiment and embodiment 5 is that the amount of water added to Radix Ophiopogonis is adjusted from 240g to 480g, and the amount of water added to the mixture of Scrophulariae and Radix Rehmanniae is adjusted from 540g to 1080g. The remaining steps remain unchanged.

[0068] Example 10

[0069] The difference between this embodiment and embodiment 5 is that the amount of water added to Radix Ophiopogonis is adjusted from 240g to 360g, and the amount of water added to the mixture of Scrophulariae and Radix Rehmanniae is adjusted from 540g to 810g. The remaining steps remain unchanged.

[0070] (1) Determination of rehmannia glutinose content

[0071] The content of rehmannia glycoside D in the enriched lyophilized powder was determined by high performance liquid chromatography (HPLC) with reference to the "Chinese Pharmacopoeia" (2020 edition).

[0072] The chromatographic conditions were as follows: octadecylsilane bonded silica gel as the filler; methanol-0.1% phosphoric acid solution (5:95) as the mobile phase; detection wavelength at 203 nm, injection volume of 10 μL, flow rate of 0.2 ml / min. The theoretical plate number calculated based on the rehmannia glutinosa D peak should be no less than 5000.

[0073] Preparation of reference solution: Take an appropriate amount of Rehmannia glutinosin D reference substance and add 25% methanol to make a reference solution containing 50 μg per 1 ml.

[0074] Preparation of test solution: Take 3 g of appropriate amount of lyophilized powder with increased liquid content and weigh accurately; place in a stoppered conical flask, accurately add 25 ml of 25% methanol, weigh the weight, heat under reflux and extract for 1 hour, take out, cool to room temperature, weigh again, add appropriate amount of methanol solution to make up the reduced weight, shake well, filter, and take the filtrate.

[0075] Determination: Pipette 10 μL of reference substance and test substance solution respectively, inject into HPLC instrument, determine, and calculate.

[0076] (2) Determination of moisture content

[0077] Determine the moisture content of the enriched lyophilized powder using the Fischer-Tropsch method. The specific steps are as follows: Prepare the enriched lyophilized powder prepared using different methods and set aside. Use a blank calibration to calibrate the moisture meter to an anhydrous state. Accurately weigh 10 mg of purified water, calibrate the moisture meter, and calculate the titer. Accurately weigh an appropriate amount of the test sample lyophilized powder (consume approximately 1 ml of commercially available Fischer-Tropsch test solution) and directly measure the moisture content of the test sample lyophilized powder using the moisture meter. Repeat this test three times and calculate the average value.

[0078] (3) Clarity determination

[0079] With reference to the "Chinese Pharmacopoeia" (2020 edition), the clarity of the enriched lyophilized powder solution was determined visually. Take the enriched lyophilized powder prepared under different methods and dissolve it in water to make the test solution. Place it in a paired turbidimetric glass tube with an equal amount of turbidity standard solution, place it vertically under a lamp in a dark room, and inspect it under strong light. In terms of clarity, level 1 is clear and translucent, level 2 is clear with a small amount of precipitate that is easily dispersed by shaking, and level 3 is clear with a small amount of precipitate that does not disperse by shaking. Level 1 is better than level 2, and level 2 is better than level 3.

[0080] The properties of the rehydrated lyophilized compositions obtained in Examples 6-10 are shown in Table 7 below.

[0081] Table 7

[0082] serial number Product Appearance Rehmannia glutinosa content (%) Moisture content (%) Clarity Example 5 Light brown powder 0.45 1.2 Level 1 Example 6 Brown powder and granules 0.34 3.0 Level 1 Example 7 Brown powder and granules 0.45 2.5 Level 2 Example 8 Brown powder and granules 0.40 2.2 Level 2 Example 9 Brown powder and granules 0.35 2.0 Level 1 Example 10 Brown powder and granules 0.40 2.2 Level 2

[0083] From the results in Table 7 above, it can be seen that after sufficient alcohol precipitation of the Scrophularia ningpoensis / Rehmannia glutinosa extract, the obtained liquid-enriched freeze-dried composition has good quality.

[0084] (IV) Stability evaluation of the lyophilized composition

[0085] The rehydration-enhanced freeze-dried compositions obtained in Examples 5, 6, and 9 and the rehydration-enhanced soup stock solutions obtained in the corresponding examples were respectively taken and compared in stability using an accelerated stability test method.

[0086] Accelerated test conditions: temperature 40℃±2℃, relative humidity 75%±5%, accelerated inspection for 3 months.

[0087] The results are shown in Table 8 below. In terms of clarity, Grade 1 is clear and translucent, Grade 2 is clear with a small amount of sediment that is easily dispersed, and Grade 3 is clear with a small amount of sediment that does not disperse. Grade 1 is better than Grade 2, and Grade 2 is better than Grade 3.

[0088] Table 8

[0089]

[0090]

[0091] From the results in Table 8 above, it can be seen that the rehydration-enhanced freeze-dried composition has better quality stability than the rehydration-enhanced soup stock solution. After storage, the clarity and rehmannia glycoside content of the rehydration-enhanced freeze-dried composition are relatively stable.

[0092] (V) Investigation of storage conditions of the lyophilized composition

[0093] The rehydration-enhanced freeze-dried composition obtained in Example 5 and the rehydration-enhanced soup stock solution obtained in the corresponding example were stored at different temperatures (20° C., 4° C., and −20° C.) for 6 months to compare the stability of their properties.

[0094] The results are shown in Table 9 below. In terms of clarity, Grade 1 is clear and translucent, Grade 2 is clear with a small amount of sediment that is easily dispersed, and Grade 3 is clear with a small amount of sediment that does not disperse. Grade 1 is better than Grade 2, and Grade 2 is better than Grade 3.

[0095] Table 9

[0096]

[0097] As shown in Table 9, the enhanced lyophilized composition group maintained stable properties within 6 months of storage at room temperature without refrigeration or freezing compared to the enhanced broth stock solution, whereas the enhanced broth stock solution deteriorated in quality after 10 days at room temperature.

[0098] (VI) Evaluation of the therapeutic effect of the enriched freeze-dried composition - α-glucosidase inhibition activity test

[0099] (1) Preparation of solution

[0100] Phosphate buffer solution (PBS buffer) with pH = 6.8: Take 250 ml of 0.2 mol / L potassium dihydrogen phosphate solution, add 118 mL of 0.2 mol / L sodium hydroxide solution, dilute to 1000 ml with pure water, and shake well.

[0101] α-glucosidase solution: Take an appropriate amount of α-glucosidase powder, dissolve it in PBS buffer and make up to volume to prepare a 2U / ml stock solution, and dilute it to 0.5U / ml with PBS buffer.

[0102] Substrate PNPG solution: Accurately weigh 188.31 mg of p-nitrophenyl-β-D-pyranoglucopyranoside (PNPG), first add 1 ml of DMSO to dissolve it, then dilute to 25 ml with PBS buffer to prepare a 25 mmol / L stock solution, and then dilute to 10 mmol / L with PBS buffer.

[0103] Sodium carbonate solution: Accurately weigh 5.30 g of sodium carbonate and dissolve it in 250 ml of pure water to prepare a 0.2 mol / L sodium carbonate solution.

[0104] Positive control group acarbose solution: accurately weigh 5.04 mg of acarbose, dissolve it in pure water and dilute to 5 ml to prepare a 1000 mg / L stock solution, which was then diluted with pure water to different concentrations: 50 μg / ml, 100 μg / ml, 200 μg / ml, 300 μg / ml, 400 μg / ml, and 500 μg / ml.

[0105] Sample solution of the enriched lyophilized composition: The enriched lyophilized composition prepared in Example 5 was dissolved in pure water and diluted to 5 mL to prepare a mother solution with a concentration of 1000 mg / L, which was then diluted with pure water to different concentrations: 50 μg / ml, 100 μg / ml, 200 μg / ml, 300 μg / ml, 400 μg / ml, and 500 μg / ml.

[0106] Sample enrichment decoction stock solution: The enrichment decoction stock solution prepared in Example 5 was diluted with pure water and fixed to 5 mL to prepare a mother solution with a concentration of 1000 mg / L, which was then diluted with pure water to different concentrations: 50 μg / ml, 100 μg / ml, 200 μg / ml, 300 μg / ml, 400 μg / ml, and 500 μg / ml.

[0107] (2) Experimental methods

[0108] In a 96-well plate, PBS solution, PNPG solution and other solutions were added in sequence as shown in Table 10 below. After mixing, the mixture was reacted in a 37°C water bath for 10 minutes. After the reaction was completed, the plate was taken out and an α-glucosidase solution in a 37°C water bath was added as shown in Table 10 below. The mixture was thoroughly mixed and reacted in a 37°C water bath for 20 minutes. Sodium carbonate solution was added to terminate the reaction. Since PNPG can be hydrolyzed to produce glucose and PNP under the action of α-glucosidase, the absorbance of PNG at 405nm was determined by UV-visible spectroscopy, and the inhibition rate of α-glucosidase of each sample was calculated. The results are shown in Table 10 below, where the blank group was only PBS solution and the positive control group was acarbose.

[0109] Table 10

[0110]

[0111] (3) Experimental results

[0112] Table 11 shows the results of α-glucosidase inhibition activity of the lyophilized composition and the broth at different concentrations. The lyophilized composition (300 μg / mL) exhibited an α-glucosidase inhibition rate of 88.5%, significantly higher than the 75.2% of the broth, and comparable to the 92.3% of the acarbose control group. This result demonstrates that the lyophilized composition exhibits a stronger inhibitory effect on α-glucosidase in vitro than the broth, helping to lower blood sugar levels.

[0113] Table 11

[0114]

[0115] (7) Animal experiments

[0116] (1) Experimental animals: Male SD rats weighing 180-200 g were fed a high-fat diet for 8 weeks to establish an obesity model. A diabetes model was further induced by a single intraperitoneal injection of 30 mg / kg of streptozotocin (STZ). The model was considered successful if the fasting blood glucose concentration of the rats was not less than 11.1 mmol / L for two consecutive measurements. The diabetic model rats were randomly divided into two groups, with 10 rats in each group.

[0117] (2) Experimental groups: model control group, enhanced liquid freeze-dried composition group, and enhanced liquid soup stock solution group.

[0118] (3) Sample solution preparation

[0119] Model control group: Normal saline was used as a normal control, and the dosage was 5 mL / kg.

[0120] Rehydration-enhanced lyophilized composition group: Take an appropriate amount of the rehydration-enhanced lyophilized composition prepared by the method of Example 5, add pure water to make it into 50 g / L, and administer at a dosage of 300 mg / kg.

[0121] Zengye Decoction stock solution group: Take an appropriate amount of Zengye Decoction stock solution prepared by the method of Example 5, dilute it with pure water to make 50 g / L, and administer it at a dose of 300 mg / kg.

[0122] (4) Administration: Rats were gavaged once daily for 4 weeks according to the table below. The model control group received 5 mL / kg of the drug, the enhanced lyophilized composition group received 300 mg / kg of the drug, and the enhanced lyophilized decoction group received 300 mg / kg of the drug.

[0123] (5) Body weight and fasting blood glucose monitoring: The body weight of rats was measured every week during the treatment period, and the fasting blood glucose concentration of rats was measured using a blood glucose meter.

[0124] (6) Serum index detection in rats: After the 4-week experiment, rats in each group were anesthetized and fixed on the operating table. 5 ml of blood was collected from the orbital cavity. The blood samples were coagulated at room temperature for 1 h, centrifuged at 3000 rpm for 15 min at 4°C, and the supernatant was collected and stored at -80°C. Serum insulin levels were measured using the ELISA kit according to the instructions.

[0125] (7) Liver PPARγ mRNA expression: After the 4-week experiment, rats were sacrificed and liver tissues were recovered. The liver tissues were rinsed with pre-cooled saline, snap-frozen in liquid nitrogen, and stored in a -80°C refrigerator. PPARγ mRNA expression was determined by PCR.

[0126] (8) Experimental results

[0127] Fasting blood glucose results: After the four-week experiment, the fasting blood glucose (FBG) of the rats in the enhanced liquid freeze-dried composition group decreased significantly by 45.2% compared to the model control group, and was superior to the 39.5% in the enhanced liquid decoction group. This result demonstrates that the enhanced liquid freeze-dried composition of the present invention has a stronger blood glucose-lowering effect in vivo.

[0128] Serum insulin level: After the 4-week experiment, the serum insulin level of rats in the enhanced lyophilized composition group was significantly reduced, indicating that it may lower blood sugar by improving insulin sensitivity.

[0129] Liver PPARγ mRNA expression: After the four-week experiment, PPARγ mRNA expression in the liver of rats treated with the enhanced liquid freeze-dried composition increased significantly by 2.1-fold, while it only increased by 1.2-fold in the enhanced liquid decoction group. There was no significant change in the model control group. PPARγ is a nuclear receptor transcription factor involved in regulating insulin sensitivity. Its increased expression may help improve insulin resistance in diabetic rats.

[0130] The results are shown in Table 12 below.

[0131] Table 12

[0132]

[0133] Therefore, the rehydration-enhanced freeze-dried composition of the present invention has more stable quality and better blood sugar-lowering effect than the original rehydration soup.

[0134] As described above, the basic principles, main features, and advantages of the present invention are shown and described. Those skilled in the art should understand that the present invention is not limited to the above-described embodiments. The above-described embodiments are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. In other words, equivalent changes and modifications made within the scope of the present invention and the contents of the specification should still fall within the scope of the present invention. The scope of protection claimed in the present invention is defined by the appended claims and their equivalents.

Claims

1. A liquid-enriched freeze-dried composition, characterized in that: It is obtained by vacuum freeze-drying the original solution of the enriched decoction.

2. The lyophilized composition according to claim 1, wherein The fluid-increasing soup stock solution is composed of ophiopogon japonicus extract and Scrophularia ningpoensis / Rehmannia glutinosa extract.

3. The rehydration-enhanced freeze-dried composition according to claim 2, characterized in that The lily pogon extract is obtained by extracting lily pogon with water.

4. The liquid-enhanced freeze-dried composition according to claim 2, characterized in that The relative density of the ophiopogon japonicus extract at 60° C. is 1.15-1.

18.

5. The liquid-enhanced freeze-dried composition according to claim 2, characterized in that The Scrophulariaceae / Rehmannia glutinosa extract is obtained by extracting Scrophulariaceae and Rehmannia glutinosa with water and precipitating with alcohol.

6. The liquid-enhanced freeze-dried composition according to claim 2, characterized in that The relative density of the Scrophulariaceae / Rehmannia glutinosa extract at 60° C. is 1.17-1.

20.

7. The rehydration-enhanced freeze-dried composition according to claim 3 or 5, characterized in that: The water extraction is performed by decocting the mixture 1-3 times, with each decocting time being 0.5-2 hours.

8. The rehydration-enhanced freeze-dried composition according to claim 1, characterized in that The vacuum freeze drying comprises the steps of freezing, vacuum sublimation and drying; The sublimation temperature is no more than 0°C.

9. The rehydration-enhanced freeze-dried composition according to claim 8, characterized in that The drying temperature is 30-55°C.

10. Use of the rehydration-enhanced freeze-dried composition according to any one of claims 1 to 9, characterized in that: As a component of a medicine or pharmaceutical composition for treating diabetes and / or promoting the recovery of pancreatic islet cell function.

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