阅读说明：本技术 一种药物组合物及其制备方法和应用 (Pharmaceutical composition and preparation method and application thereof ) 是由 贾雯靖 毕齐茂 赵晓辉 岳会兰 陶吉红 刘丽莹 于 2021-09-10 设计创作，主要内容包括：一种药物组合物及其制备方法和应用,涉及中药组合物技术领域,其该药物组合物中含有60％乙醇提取的党参提取物,制备方法包括浸提：取10～50kg党参,切成2～5cm段,用乙醇冷浸提取,料液比为1:10,提取1～3次,每次一天,混合几次提取液浓缩干燥,得到乙醇提取的浸膏；得到的60％乙醇提取的浸膏用纯水溶解后上D-101大孔吸附树脂柱脱糖；脱糖后的溶液再用60％的乙醇洗脱；洗脱后浓缩干燥得到60％乙醇提取的党参提取物。本发明的有益效果在于：60％乙醇提取的党参提取物具有较好的降低餐后血糖效果。(A pharmaceutical composition and a preparation method and application thereof relate to the technical field of traditional Chinese medicine compositions, wherein the pharmaceutical composition contains a codonopsis pilosula extract extracted by 60% ethanol, and the preparation method comprises the following steps: taking 10-50 kg of codonopsis pilosula, cutting into sections of 2-5 cm, and performing cold soaking extraction by using ethanol, wherein the material-liquid ratio is 1:10, extracting for 1-3 times, mixing extracting solutions for several times each day, concentrating and drying to obtain an ethanol-extracted extract; dissolving the obtained extract with 60% ethanol with pure water, and removing sugar with D-101 macroporous adsorbent resin column; eluting the desugarized solution with 60% ethanol; eluting, concentrating, and drying to obtain radix Codonopsis extract extracted with 60% ethanol. The invention has the beneficial effects that: the codonopsis pilosula extract extracted by 60% ethanol has better effect of reducing postprandial blood sugar.)

1. A pharmaceutical composition characterized by: the medicinal composition contains radix Codonopsis extract.

2. A pharmaceutical composition according to claim 1, wherein: the radix Codonopsis extract is ethanol extracted radix Codonopsis extract.

3. A pharmaceutical composition according to claim 2, wherein: the ethanol extraction is 20-95% ethanol extraction.

4. A pharmaceutical composition according to claim 3, wherein: the preparation method of the codonopsis pilosula extract extracted by ethanol comprises the following steps:

step one, leaching: taking 10-50 kg of codonopsis pilosula, cutting into sections of 2-5 cm, and performing cold soaking extraction by using ethanol, wherein the material-liquid ratio is 1:10, extracting for 1-3 times, mixing extracting solutions for several times each day, concentrating and drying to obtain an ethanol-extracted extract;

step two, desugaring: dissolving the obtained ethanol extract with pure water, and removing sugar with D-101 macroporous adsorbent resin column;

step three, elution: eluting the desugarized solution with ethanol;

step four, concentrating and drying: eluting, concentrating, and drying to obtain radix Codonopsis extract extracted with ethanol.

5. A pharmaceutical composition according to claim 4, wherein: the application of the pharmaceutical composition containing the codonopsis pilosula extract in preparing hypoglycemic drugs.

6. The use of the pharmaceutical composition containing Codonopsis Pilosulae extract as claimed in claim 5, wherein: the hypoglycemic drug can be prepared into tablets, pills, capsules, granules or oral liquid.

7. The use of the pharmaceutical composition containing Codonopsis Pilosulae extract as claimed in claim 5 in the preparation of hypoglycemic drugs, characterized in that: the activity of the codonopsis pilosula extract of the 60 percent ethanol extraction part in the 20 percent to 95 percent ethanol for reducing blood sugar is higher than that of the codonopsis pilosula extract of other ethanol extraction parts with concentration.

Technical Field

The invention relates to the technical field of traditional Chinese medicine compositions, in particular to a pharmaceutical composition and a preparation method and application thereof.

Background

Blood sugar is the only standard for diagnosing diabetes, which can be diagnosed by fasting blood sugar greater than or equal to 7.0 mmol/L and/or blood sugar greater than or equal to 11.1 mmol/L after two hours, and there is no method for radically treating diabetes at present, but diabetes can be controlled well by various therapeutic means, and there are many medicines for controlling blood sugar activity such as acarbose, which is a glucosidase inhibitor, complex oligosaccharide, and the structure of which is similar to oligosaccharide. Acarbose, also known as bayer oxabetine, is used mainly in the treatment of diabetes. Diabetes, a disease of which is currently a disease of abundance, requires more research and drug development. The codonopsis pilosula is often mentioned in a traditional Chinese medicine formula for treating diabetes, but the codonopsis pilosula extract has no detailed research report on the aspects of treating diabetes and reducing blood sugar.

Disclosure of Invention

Aiming at the problems, the invention provides a pharmaceutical composition, a preparation method and application thereof, which further research the effect of the codonopsis pilosula extract on reducing the activity of blood sugar and solve the application problem of the codonopsis pilosula extract in the aspects of reducing the blood sugar and treating diabetes.

The invention provides a pharmaceutical composition, a preparation method and application thereof, wherein the pharmaceutical composition contains a codonopsis pilosula extract.

Further, the radix codonopsitis extract is an ethanol-extracted radix codonopsitis extract.

Further, the ethanol extraction is 20-95% ethanol extraction.

Further, the preparation method of the codonopsis pilosula extract extracted by ethanol comprises the following steps:

step one, leaching: taking 10-50 kg of codonopsis pilosula, cutting into sections of 2-5 cm, and performing cold soaking extraction by using ethanol, wherein the material-liquid ratio is 1:10, extracting for 1-3 times, mixing extracting solutions for several times each day, concentrating and drying to obtain an ethanol-extracted extract;

step two, desugaring: dissolving the obtained ethanol extract with pure water, and removing sugar with D-101 macroporous adsorbent resin column;

step three, elution: eluting the desugarized solution with ethanol;

step four, concentrating and drying: eluting, concentrating, and drying to obtain radix Codonopsis extract extracted with ethanol.

Furthermore, the application of the pharmaceutical composition containing the codonopsis pilosula extract in preparing hypoglycemic drugs.

Further, the hypoglycemic drug can be prepared into tablets, pills, capsules, granules or oral liquid.

Furthermore, the activity of the codonopsis pilosula extract of the 60% ethanol extraction part in the 20-95% ethanol for reducing blood sugar is higher than that of the codonopsis pilosula extract of other ethanol extraction parts with concentration.

The invention has the beneficial effects that: the codonopsis pilosula extract extracted by ethanol has constructive help for research on reducing postprandial blood sugar, and has great effects in clinical research and pharmacy, particularly, experiments prove that the codonopsis pilosula extract extracted by 60% ethanol has better effect of reducing postprandial blood sugar.

Drawings

FIG. 1 is a graph showing the change in the α -glucosidase inhibitory activity of the present invention;

FIG. 2 is a graph showing the change in the sucrase inhibitory activity of the present invention;

FIG. 3 is a graph showing the change in the inhibitory activity of maltase according to the present invention;

FIG. 4 is a graph showing the change in the amylase inhibitory activity of the present invention;

FIG. 5 is a graph of oral starch tolerance glycemic variability in diabetic mice of the present invention;

FIG. 6 is a graph of oral sucrose tolerance glycemic variability of diabetic mice of the present invention;

fig. 7 is a graph showing the oral maltose tolerance glucose change in diabetic mice of the present invention.

Detailed Description

Example 1

The invention provides a pharmaceutical composition, a preparation method and application thereof, wherein the pharmaceutical composition contains a codonopsis pilosula extract.

Further, the radix codonopsitis extract is an ethanol-extracted radix codonopsitis extract.

Further, the ethanol extraction is 20% ethanol extraction.

Further, the preparation method of the codonopsis pilosula extract extracted by ethanol comprises the following steps:

step one, leaching: cutting 20kg of radix Codonopsis into 3cm sections, extracting with ethanol at a material-liquid ratio of 1:10 for 3 times, concentrating and drying the extractive solution 3 to obtain ethanol extract;

step two, desugaring: dissolving the obtained ethanol extract with pure water, and removing sugar with D-101 macroporous adsorbent resin column;

step three, elution: eluting the desugarized solution with ethanol;

step four, concentrating and drying: eluting, concentrating, and drying to obtain radix Codonopsis extract extracted with ethanol.

Example 2

The invention provides a pharmaceutical composition, a preparation method and application thereof, wherein the pharmaceutical composition contains a codonopsis pilosula extract.

Further, the radix codonopsitis extract is an ethanol-extracted radix codonopsitis extract.

Further, the ethanol extraction is 60% ethanol extraction.

Further, the preparation method of the codonopsis pilosula extract extracted by ethanol comprises the following steps:

step one, leaching: cutting 20kg of radix Codonopsis into 3cm sections, extracting with ethanol at a material-liquid ratio of 1:10 for 3 times, concentrating and drying the extractive solution 3 to obtain ethanol extract;

step two, desugaring: dissolving the obtained ethanol extract with pure water, and removing sugar with D-101 macroporous adsorbent resin column;

step three, elution: eluting the desugarized solution with ethanol;

step four, concentrating and drying: eluting, concentrating, and drying to obtain radix Codonopsis extract extracted with ethanol.

Example 3

The invention provides a pharmaceutical composition, a preparation method and application thereof, wherein the pharmaceutical composition contains a codonopsis pilosula extract.

Further, the radix codonopsitis extract is an ethanol-extracted radix codonopsitis extract.

Further, the ethanol extraction is 95% ethanol extraction.

Further, the preparation method of the codonopsis pilosula extract extracted by ethanol comprises the following steps:

step one, leaching: cutting 20kg of radix Codonopsis into 3cm sections, extracting with ethanol at a material-liquid ratio of 1:10 for 3 times, concentrating and drying the extractive solution 3 to obtain ethanol extract;

step two, desugaring: dissolving the obtained ethanol extract with pure water, and removing sugar with D-101 macroporous adsorbent resin column;

step three, elution: eluting the desugarized solution with ethanol;

step four, concentrating and drying: eluting, concentrating, and drying to obtain radix Codonopsis extract extracted with ethanol.

Example 4

As shown in figures 1-7, the invention provides a pharmaceutical composition, a preparation method and an application thereof, wherein the pharmaceutical composition contains a codonopsis pilosula extract.

The hypoglycemic activity of the codonopsis pilosula extracts extracted by 20%, 60% and 95% ethanol is respectively detected by in vitro experiments and in vivo experiments.

Further, the in vitro experiment comprises screening of alpha-glucosidase inhibitory activity, screening of sucrase inhibitory activity, screening of maltase inhibitory activity, screening of alpha-amylase inhibitory activity, extraction of rat small intestinal glycosidase and activity determination; the in vivo experiment comprises an oral starch tolerance experiment of a diabetic mouse, an oral sucrose tolerance experiment of the diabetic mouse and an oral maltose tolerance experiment of the diabetic mouse.

Further, the screening method for the alpha-glucosidase inhibitory activity comprises the following steps: adding 50 μ L of 5mg/mL radix Codonopsis extract sample and 50uL of 0.5U/mL enzyme solution into 96-well plate under ice bath condition, placing in 37 deg.C constant temperature oscillator, incubating at 100r/min for 10min, and further performing ice bathAdding 50uL0.5 mmol/mL p-NPG, placing in a constant temperature oscillator at 37 deg.C, incubating at 100r/min for 20min, immediately putting in ice water bath for 5min, reducing enzyme activity, and continuously adding 0.1mol/L Na₂CO₃The reaction was stopped with 50. mu.L of the solution and repeated three times; the detection was carried out at a wavelength of 409nm and the inhibitory activity of the sample on the enzyme was calculated, as shown in FIG. 1.

Further, the screening method for sucrase inhibitory activity comprises the following steps: adding 50 μ L of enzyme solution containing 17.5U/mL and 50 μ L of 5mg/mL radix Codonopsis extract sample into 48-well plate, and pre-incubating at 37 deg.C for 10 min; then 50. mu.L of 0.5mol/L sucrose solution was added and the mixture was incubated at 37 ℃ for 20 min; immediately putting into ice water bath for 5min, reducing enzyme activity, and continuously adding 0.1mol/LNa₂CO₃The reaction was stopped with 50. mu.L of the solution and repeated three times; the glucose concentration was determined using a glucose kit and the sucrase inhibitory activity of the sample was calculated as shown in FIG. 2.

Further, the screening method for the maltase inhibitory activity comprises the following steps: adding 50 μ L of enzyme solution containing 11.56U/mL and 50 μ L of 5mg/mL radix Codonopsis extract sample into 48-well plate, and pre-incubating at 37 deg.C for 10 min; then, 50. mu.L of 1.39mmol/mL maltose solution was added and the mixture was incubated at 37 ℃ for 20 min; immediately putting into ice water bath for 5min, reducing enzyme activity, and continuously adding 0.1mol/L Na₂CO₃The reaction was stopped with 50. mu.L of the solution and repeated three times; the glucose concentration was determined using a glucose kit and the inhibitory activity of the samples on maltase was calculated as shown in fig. 3.

Further, the screening method for the alpha-amylase inhibitory activity comprises the following steps: under the ice bath condition, 50 mu L of 5mg/mL codonopsis pilosula extract sample and 50 mu L of 10U/mL enzyme solution are added into a 96-well plate, put into a constant temperature oscillator at 37 ℃, incubated for 10min at 100r/min, 50 mu L of 0.1% (w/v) starch is added under the ice bath condition, then put into a constant temperature oscillator at 37 ℃, incubated for 20min at 100r/min, immediately put into an ice water bath for 5min, the enzyme activity is reduced, 100 mu L of LDNS solution is continuously added, reacted for 5min at 100 ℃, and the reaction is repeated for three times; after cooling to room temperature, the measurement was carried out at a wavelength of 540nm, and the inhibitory activity of the sample on the enzyme was calculated, as shown in FIG. 4.

Further, the method for extracting and determining the activity of the rat small intestinal glycosidase comprises the following steps: after fasting for 12h, rats are killed by cervical vertebra removal, the small intestine is immediately taken out and placed on an ice bench, the small intestine is cut open and the intestinal mucosa is exposed in a turning way, washed by PBS precooled at 4 ℃ and then wiped dry, the intestinal mucosa is scraped by a glass slide, and the weight-volume ratio of the small intestine to the total weight of the rat is 1: 5 adding into 4 deg.C precooled PBS, homogenizing, centrifuging at 4 deg.C 8000r/min for 20min, collecting supernatant, packaging, and storing at-20 deg.C; taking 0.1mL of supernatant enzyme solution, adding 0.35mL of PBS, carrying out water bath at 37 ℃ for 10min, adding 50 mu L of 0.25mol/L sucrose or maltose solution, immediately putting into ice water bath for 5min after 20min, and continuously adding 0.5mL of Na2CO3 solution to terminate the reaction; 3-hole parallel determination, wherein 1 mu moL of glucose generated in 1L of solution per minute at 37 ℃ and pH 6.8 is defined as 1 enzyme activity unit; the enzyme activity is equal to the sugar concentration x 2 x 1000/20.

Furthermore, in the in vitro experiment, the alpha-glucosidase inhibition activities of the codonopsis pilosula extracts extracted by 20%, 60% and 95% ethanol are compared, and the inhibition activities of the codonopsis pilosula extracts extracted by 20% and 60% ethanol are higher, and the inhibition rates are all more than 80%; and the IC50 values of the extracts of 20%, 60% and 95% of the codonopsis pilosula and the acarbose which is a positive drug are measured, and the results show that the IC50 values of the sucrases of the extracts of 20%, 60% and 95% of the codonopsis pilosula are respectively 0.1383, 0.2413 and 0.7169mg/mL, the IC50 value of the acarbose which is the positive drug is 0.4634 mu g/mL, and compared with the IC50 value of the positive control drug, the sucrase inhibitory activity of the extracts of 20%, 60% and 95% of the codonopsis pilosula is better than that of the acarbose; maltose IC50 values of 20%, 60% and 95% of radix Codonopsis extract are 0.2318, 0.3258 and 1.208mg/mL respectively, acarbose IC50 value of positive drug is 0.2525 μ g/mL, and maltose of 20% and 60% ethanol part of radix Codonopsis has inhibitory activity compared with IC50 value of positive control drug.

Further, the oral starch tolerance test method for the diabetic mice comprises the following steps: diabetic mice were fasted for 12h and randomly divided into 4 groups of 10 mice each; group 1 was a control group: perfusing the diabetic mice with 3g/kg of starch; group 2 is a positive group: perfusing the diabetic mice with a mixture of 8mg/kg acarbose and 3g/kg starch; groups 3-4 are experimental groups: perfusing the diabetic mice with 200mg/kg and 600mg/kg samples extracted with 60% ethanol and 3g/kg starch mixture, respectively; blood was taken from the tail vein at 0, 30, 60 and 120min and blood glucose concentration was measured using a glucometer, and a postprandial blood glucose change curve was plotted by prism7.0 and the area under the blood glucose level curve was calculated, as shown in fig. 5.

Further, the oral sucrose tolerance test method for the diabetic mice comprises the following steps: diabetic mice were fasted for 12h and randomly divided into 4 groups of 10 mice each; group 1 was a control group: perfusing the diabetic mice with 3g/kg of sucrose; group 2 is a positive group: gavage the diabetic mice with a mixture of 8mg/kg acarbose and 3g/kg sucrose; groups 3-4 are experimental groups: gavage the diabetic mice with 200mg/kg and 600mg/kg of 60% ethanol-extracted samples and 3g/kg of sucrose mixture, respectively; blood was taken from the tail vein at 0, 30, 60 and 120min and blood glucose concentration was measured using a glucometer, and a postprandial blood glucose change curve was plotted by prism7.0 and the area under the blood glucose level curve was calculated, as shown in fig. 6.

Further, the oral maltose tolerance test method for the diabetic mice comprises the following steps: diabetic mice were fasted for 12h and randomly divided into 4 groups of 10 mice each; group 1 was a control group: perfusing the diabetic mice with 3g/kg maltose; group 2 is a positive group: gavage diabetic mice with a mixture of 8mg/kg acarbose and 3g/kg maltose; groups 3-4 are experimental groups: gavage diabetic mice with 200mg/kg and 600mg/kg of 60% ethanol-extracted samples and 3g/kg of maltose mixture, respectively; blood was taken from the tail vein at 0, 30, 60 and 120min and blood glucose concentration was measured using a glucometer, and a postprandial blood glucose change curve was plotted by prism7.0 and the area under the blood glucose level curve was calculated, as shown in fig. 7.

Furthermore, the in vivo experiment selects 60% ethanol extract of codonopsis pilosula with strong in vitro activity to perform the activity experiment of reducing postprandial blood sugar in a diabetic mouse, the tolerance test of starch, sucrose and maltose of the diabetic mouse is carried out, the dosages are all 3g/kg, the postprandial blood sugar reducing effects of the 60% ethanol extract of codonopsis pilosula with dosages of 200mg/kg and 600mg/kg and the 8mg/kg positive drug acarbose are evaluated, and the experiment result shows that the 60% ethanol extract of codonopsis pilosula has a good effect of reducing the postprandial blood sugar of the mouse.

Example 5

The hypoglycemic medicine containing Codonopsis pilosula extract extracted with 60% ethanol is made into tablet.

Example 6

The hypoglycemic medicine containing the codonopsis pilosula extract extracted by 60% ethanol is prepared into pills.

Example 7

The hypoglycemic medicine containing the codonopsis pilosula extract extracted by 60% ethanol is prepared into capsules.

Example 8

The hypoglycemic medicine containing the codonopsis pilosula extract extracted by 60% ethanol is prepared into granules.

Example 9

The hypoglycemic medicine containing the codonopsis pilosula extract extracted by 60% ethanol is prepared into oral liquid.