Hedera helix extract as well as preparation method and application thereof

Abstract

The preparation method comprises the following steps: extracting hedera helix leaves with 75%-85% ethanol, concentrating, adjusting the pH value to 9.5-11, standing, filtering to obtain a crude extract, passing the crude extract through a chromatographic column, and eluting with 60%-80% ethanol to obtain the hedera helix extract; the chromatographic column is an HPD100 macroporous resin column, a polyamide column or a C18 column. The invention also provides pharmaceutical application of the hedera helix extract. By optimizing the preparation process, anti-inflammatory components in the hedera helix raw material are enriched, the yield of the obtained extract is increased, the saponin content is reduced, but the anti-inflammatory effect of the obtained extract is better, and an unexpected technical effect is achieved. According to the invention, the technical prejudice of increasing the content of saponin in the prior art is overcome, the hedera helix extract with better anti-inflammatory effect and higher yield is obtained on the basis of simplifying the elution procedure, the anti-inflammatory activity of the extract is increased, the anti-inflammatory effect is better under the condition of the same weight of crude drug, the dosage is reduced, and the cost is reduced. The economic benefit and the utilization rate of the raw medicines are improved.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine extraction, and in particular to an ivy extract and a preparation method and application thereof. Background Art

[0002] The ethanol extract of Hedera helix has multiple effects such as antispasmodic, mucolytic, anti-inflammatory, antibacterial and antiviral. Hedera helix preparations are currently on the market in many countries around the world and are widely used for respiratory diseases such as respiratory mucositis, chronic bronchitis, and acute respiratory inflammation with cough. At present, the ethanol extract of Hedera helix is ​​used as an expectorant and is made into tablets, capsules, effervescent tablets, oral liquids and other dosage forms, and is approved for marketing in many countries including Europe. As a botanical medicine, Hedera helix preparations have good safety and tolerability in children. The currently listed Prospan brand Hedera helix leaf extract syrup contains 7 mg / mL of Hedera helix leaf extract extracted with 30% ethanol, 1.34 mg of potassium sorbate, 500 μg of anhydrous citric acid, 550 mg of sorbitol, 1.5 mg of xanthan gum, 3 mg of essence, and pure water to 1 mL. Among them, the Hedera helix leaf extract is a crude extraction of 30% ethanol, and the content of saponins and other active ingredients is low.

[0003] The main active substances in Hedera helix are saponins, triterpenoid saponins disaccharide glycosides include hedera saponin B, hedera saponin C, hedera saponin D, and monosaccharide glycoside α-hedera saponin. Other ingredients include flavonoids, coumarins, polyacetylene, phenolic acids, alkaloids, volatile oils, etc.

[0004] The prior art discloses a variety of methods for preparing Hedera helix extracts.

[0005] The one-test-multiple-evaluation method was used to simultaneously determine four saponins in ivy oral liquid (Chinese Patent Medicine, Vol. 40, No. 8, 2018). The solution was extracted with 80% ethanol, treated with alkali solution, and allowed to stand overnight at 4°C. The filtrate was passed through a DM130 macroporous resin column and gradient eluted with 30% ethanol and 70% ethanol. The 70% ethanol eluate was collected and concentrated to obtain the extract. However, the anti-inflammatory effect of the extract, the extract yield, the saponin content, etc. were not disclosed.

[0006] The abstract of the Optimization of Preparation Process of Hedera Helichrysum Extract (Yang Ziwen, 2018) states that 80% ethanol was used for extraction, 1% of the amount of medicinal materials was added for alkali treatment, and the extract was precipitated at 4°C for 8 hours. The filtrate was passed through an HPD-100 macroporous resin column, and gradient eluted with 30% ethanol and 80% ethanol. The 80% ethanol eluate was collected and concentrated to obtain the extract. This method mainly optimized the extraction content and recovery rate of Hedera Helichrysum Saponin C and α-Ivy Saponin, but did not involve other saponin components such as Hedera Helichrysum Saponin B and Ivy Saponin D, and the extract was not disclosed. However, the document also did not disclose the extract yield and the anti-inflammatory effect of the extract.

[0007] CN104306416A discloses a drug for treating bronchitis, pneumonia and asthma and its preparation method (publication date: 2015-01-28), wherein the preparation method of ivy saponin components is recorded: weigh the stems and leaves of ivy, reflux extract with 10-90% purity ethanol solution in an extraction kettle according to the mass volume ratio of ivy stems and leaves to ethanol solution of 1:3-10, filter after reflux extraction, centrifuge, concentrate under reduced pressure, and evaporate to extract; dissolve with deionized water, pass through a macroporous resin column, and elute with water and ethanol solution in turn; concentrate the ethanol eluate under reduced pressure, dry under vacuum at 60°C, weigh, and obtain ivy saponin. The specification discloses the effect of ivy oral liquid made of ivy saponin and ivy volatile oil on the SOD activity in serum, lung tissue and BALF of rats with chronic bronchitis, but does not disclose the anti-inflammatory effect of ivy saponin components used alone.

[0008] CN102188465A discloses a preparation method and use of an ivy plant extract (publication date: 2011-09-21), wherein the ivy extract is extracted with an extraction solvent, the extract is subjected to a macroporous adsorption resin, and the alcohol eluent is dried to obtain an extract, and the total saponin content in the extract is not less than 30%. The extract can be used alone or in combination with other extracts to prepare drugs for treating rheumatoid arthritis, respiratory diseases, hepatitis, etc. However, the specification only uses the enrichment of total saponin content as a condition optimization target, and does not disclose the relationship between total saponins, saponin C and α-saponin and drug efficacy.

[0009] In the prior art, the extraction of ivy mainly focuses on increasing the content and recovery rate of saponins. For example, the use of 30% ethanol + 70% ethanol gradient elution is to remove impurities with high polarity, such as polysaccharides, pigments, etc., in order to enrich saponins in ivy extracts and increase the saponin content. However, the prior art has not studied whether the anti-inflammatory efficacy of saponins is related to their content or recovery rate. From the perspective of common knowledge, the higher the saponin content, the better the efficacy should be. However, the applicant has found through research that there may be a phenomenon that is contrary to common knowledge. The prior art has not established a connection between the amount of alkali solution used in the extraction process, the relationship between the elution gradient and the anti-inflammatory efficacy, and has not carried out corresponding research. It is necessary to further optimize the extraction process from the perspective of improving the anti-inflammatory efficacy of ivy extracts. Summary of the invention

[0010] The purpose of the present invention is to solve the technical problems in the prior art that the ivy leaf extract has unknown components and poor anti-inflammatory effect.

[0011] The present invention provides an ivy extract with a simpler preparation process, certain ingredients and better anti-inflammatory effect. The present invention improves the yield of soluble effective ingredients by optimizing the extraction process, improves the anti-inflammatory efficacy of the extract at a lower saponin content, shortens the production cycle and saves production costs.

[0012] The technical solution adopted by the present invention is:

[0013] A method for preparing an ivy extract, comprising: extracting ivy leaves with 75% to 85% ethanol, concentrating, adjusting the pH to 9.5 to 11, standing, filtering, passing the obtained crude extract through a chromatography column, and eluting with 60% to 80% ethanol to obtain the ivy extract; the chromatography column is one of an HPD100 macroporous resin column, a polyamide column or a C18 column.

[0014] Furthermore, when the chromatography column is a polyamide column, the pH of the crude extract is preferably adjusted to neutral before passing through the chromatography column.

[0015] Furthermore, when the chromatography column is a C18 column, the crude extract can be eluted with 30% ethanol and then with 60% to 80% ethanol when passing through the chromatography column, and the eluate eluted with 60% to 80% ethanol is collected.

[0016] Furthermore, it is preferred that the chromatography column is a HPD100 macroporous resin column.

[0017] Further, the method comprises the following steps:

[0018] (1) Take ivy leaves, crush them, extract them with 75% to 85% ethanol under reflux, and filter and combine the filtrate;

[0019] (2) The filtrate is concentrated until there is no alcohol taste, the pH is adjusted to 9.5 to 11, refrigerated and allowed to stand for 8 to 15 hours, and filtered to obtain a crude extract;

[0020] (3) The crude extract is passed through a chromatography column, eluted with water until colorless, and then eluted with 3 to 5 times the column volume of 60% to 80% ethanol. The ethanol eluate is collected, concentrated and dried to obtain the ivy extract; the chromatography column is one of an HPD100 macroporous resin column, a polyamide column or a C18 column.

[0021] In the step (1), the reflux extraction can be performed multiple times, such as 2 to 4 times, preferably 2 to 3 times.

[0022] In the step (1), the volumetric dosage of 75% to 85% ethanol is generally 5-12 mL / g based on the mass of ivy leaves.

[0023] In the step (1), the temperature of the reflux extraction is preferably 80 to 90°C.

[0024] In the step (3), when the chromatography column is a polyamide column, the pH of the crude extract is preferably adjusted to neutral before passing through the chromatography column.

[0025] When the chromatography column is a C18 column, after the crude extract passes through the chromatography column, it can be eluted with 30% ethanol first, and then eluted with 3 to 5 times the column volume of 60% to 80% ethanol, and the eluate eluted with 60% to 80% ethanol is collected for concentration and drying. The elution volume of the 30% ethanol is 2 to 3 times the column volume, and this part of the eluate is discarded.

[0026] Most preferably, the chromatography column is a HPD100 macroporous resin column.

[0027] In the step (2), it is preferred to use an alkaline solution to adjust the pH value, and the alkaline solution includes a sodium hydroxide solution or a potassium hydroxide solution. After adding the alkaline solution to adjust the pH value, water can be added to a total volume of 5 to 12 times the amount of ivy leaves added (volume to mass ratio).

[0028] The concentration of the sodium hydroxide solution or potassium hydroxide solution is preferably 3-8%, more preferably 5%.

[0029] The volume dosage of the alkaline solution is preferably 0.2 to 0.4 times the dosage of the ivy leaves, preferably 0.2 to 0.3 times, and more preferably 0.3 times.

[0030] More preferably, the mass of sodium hydroxide or potassium hydroxide in the alkaline solution is 1-2% of the amount of ivy leaves added.

[0031] The ivy includes a variety of medicinal materials such as Hedera helix L., Hedera nepalensis, Hedera canariensis, Hedera colchica, and Japanese ivy. Among them, Hedera helix L. and Hedera chinensis are mostly used for drug treatment.

[0032] The invention also provides ivy extract prepared according to the preparation method.

[0033] Furthermore, the ivy extract includes α-hedera saponin, hedera saponin C, hedera saponin B and hedera saponin D, and the total mass fraction of α-hedera saponin, hedera saponin C, hedera saponin B and hedera saponin D is greater than 50%.

[0034] Furthermore, in the ivy extract, the mass content of the α-hedera saponin is greater than 10%, the mass content of the hedera saponin C is greater than 30%, the mass content of the hedera saponin B is greater than 2%, and the mass content of the hedera saponin D is greater than 1%.

[0035] The present invention also provides application of the ivy extract in preparing a medicine for treating or preventing acute bronchitis.

[0036] The present invention also provides application of the ivy extract in preparing expectorant and antitussive drugs.

[0037] The invention also provides a medicine for treating or preventing acute bronchitis, comprising the ivy extract.

[0038] The invention also provides an expectorant and antitussive medicine, comprising the ivy extract.

[0039] The beneficial effects of the present invention are:

[0040] The present invention enriches the anti-inflammatory components in the ivy raw material by optimizing the preparation process, and clarifies that the ivy extract with a specific component content has a good drug effect. The present invention obtains an increase in the yield and a decrease in the saponin content in the extract, but the anti-inflammatory effect of the obtained extract is better, and an unexpected technical effect is achieved. The present invention overcomes the technical bias of the prior art to blindly increase the saponin content, and obtains an ivy extract with better anti-inflammatory effect and higher yield on the basis of simplifying the elution procedure, which is more conducive to the development of oral solution preparations and reduces the use of solubilizers. The present invention can effectively enrich the anti-inflammatory components in the ivy leaf extract, and the anti-inflammatory activity of the extract is increased. Therefore, the anti-inflammatory effect is better under the same weight of raw medicine, which is conducive to reducing the dosage and improving the economic benefits and utilization rate of the raw medicine. DETAILED DESCRIPTION

[0041] The technical solution of the present invention is further described below with reference to specific embodiments, but the protection scope of the present invention is not limited thereto.

[0042] Taking 200 g of Hedera helix leaf slices as an example, an extract of Hedera helix leaf was prepared according to the following example. The production batch number of Hedera helix medicinal material is LX220501.

[0043] Example 1: Preparation of Extract 1

[0044] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 1000 mL (5 times the volume-to-mass ratio of the medicinal material) of 85% ethanol, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add 80 mL (0.4 times the feed amount) of 5% sodium hydroxide solution, add water to the total volume of 5 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, adsorb the filtrate through the macroporous resin of HPD100, elute with water until colorless, and then elute with 5 times the column volume of 60% ethanol, collect the 60% ethanol eluate, concentrate, dry, and crush to obtain.

[0045] Example 2: Preparation of Extract 2

[0046] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 80% ethanol in a volume-to-mass ratio of 12 times that of the medicinal material, the extraction temperature is 80-90° C. Filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add 0.3 times the feed amount of 5% NaOH solution, add water to the total volume of 12 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, pass through HPD100 macroporous adsorption resin, elute with water until colorless, then elute with 70% ethanol in a volume of 3 times the column volume, collect the 70% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0047] Example 3: Preparation of Extract 3

[0048] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 75% ethanol in a volume mass ratio of 9 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add 0.2 times the feed amount of 5% NaOH solution, add water to the total volume of 9 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, pass through HPD100 macroporous adsorption resin, elute with water until colorless, then elute with 2 times the column volume of 80% ethanol, collect the 80% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0049] Example 4: Preparation of Extract 4

[0050] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 85% ethanol in a volume mass ratio of 5 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add potassium hydroxide solution, add water to the total volume of 5 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, adjust the pH of the filtrate to neutral, pass the filtrate through a polyamide column (30-60 mesh), elute with water until colorless, then elute with 60% ethanol in a volume of 5 times that of the column, collect the 60% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0051] Example 5: Preparation of Extract 5

[0052] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 80% ethanol in a volume-to-mass ratio of 12 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrates, recover the ethanol until there is no alcohol taste, use potassium hydroxide solution, add water to a total volume of 12 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, adjust the pH of the filtrate to neutral, pass the filtrate through a polyamide column (30-60 mesh), elute with water until colorless, then elute with 70% ethanol in a volume of 3 times that of the column, collect the 70% ethanol eluate, concentrate the filtrate, dry, and crush to obtain the product.

[0053] Example 6: Extract 6

[0054] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 75% ethanol in a volume mass ratio of 9 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add potassium hydroxide solution, add water to a total volume of 9 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, adjust the pH of the filtrate to neutral, pass the filtrate through a polyamide column (30-60 mesh), elute with water until colorless, then elute with 80% ethanol in a volume of 2 times the column volume, collect the 80% ethanol eluate, concentrate the filtrate, dry, and crush to obtain the product.

[0055] Example 7: Extract 7

[0056] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 85% ethanol in a volume mass ratio of 5 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add sodium hydroxide solution, add water to the total volume of 5 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, concentrate to 100-200 mL, pass through a C18 column (particle size 60 μm), elute with water until colorless, then elute with 60% ethanol in a volume of 5 times that of the column, collect the 60% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0057] Example 8: Extract 8

[0058] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 85% ethanol in a volume mass ratio of 5 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add water to the sodium hydroxide solution to a total volume of 5 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, concentrate to 100-200 mL, and then pass through a C18 column (particle size 60 μm), elute with water until colorless, then elute with 2 times the amount of 30% ethanol, and then elute with 5 times the column volume of 60% ethanol, collect the 60% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0059] Example 9: Extract 9

[0060] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 80% ethanol in a volume-to-mass ratio of 12 times that of the medicinal material, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add sodium hydroxide solution, add water to a total volume of 12 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, concentrate to 100-200 mL, pass through a C18 column (particle size 60 μm), elute with water until colorless, then elute with 70% ethanol in a volume of 3 times that of the column, collect the 70% ethanol eluate, concentrate the filtrate, dry, and crush to obtain the product.

[0061] Example 10: Extract 10

[0062] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract them twice with 80% ethanol in a volume-to-mass ratio of 12 times that of the medicinal materials, the extraction temperature is 80-90° C. Filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add sodium hydroxide solution, add water to a total volume of 12 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, concentrate to 100-200 mL, pass through a C18 column (particle size 60 μm), elute with water until colorless, then elute with 2 times the column volume of 30% ethanol, and then elute with 3 times the column volume of 70% ethanol, collect the 70% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0063] Example 11: Extract 11

[0064] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 75% ethanol in a volume mass ratio of 9 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add sodium hydroxide solution, add water to the total volume of 9 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, concentrate to 100-200 mL, pass through a C18 column (particle size 60 μm), elute with water until colorless, then elute with 80% ethanol in a volume of 2 times the column volume, collect the 80% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0065] Example 12: Extract 12

[0066] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 75% ethanol in a volume-to-mass ratio of 9 times that of the medicinal materials, the extraction temperature is 80-90° C. Filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add sodium hydroxide solution, add water to a total volume of 9 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, concentrate to 100-200 mL, pass through a C18 column (particle size 60 μm), elute with water until colorless, then elute with 2 times the column volume of 30% ethanol, and then elute with 2 times the column volume of 80% ethanol, collect the 80% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0067] Example 13: Control group 1

[0068] Based on Example 2, the alkali solution was adjusted to 1% of the medicinal material amount, a DM130 chromatographic column was used, and 30% ethanol was added for elution, as follows:

[0069] Weigh 200g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 80% ethanol in a volume-to-mass ratio of 12 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add 0.2 times the feed amount of 5% NaOH solution, add water to a total volume of 12 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, pass through DM130 macroporous adsorption resin, elute with water until colorless, then elute with 3 times the column volume of 30% ethanol, and then elute with 3 times the column volume of 70% ethanol, collect the 70% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0070] Example 14: Control group 2

[0071] Based on control group 1, HPD100 chromatographic column was used, as follows:

[0072] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 80% ethanol in a volume-to-mass ratio of 12 times that of the medicinal materials, the extraction temperature is 80-90° C. Filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add 0.2 times the feed amount of 5% NaOH solution, add water to a total volume of 12 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, pass through HPD100 macroporous adsorption resin, elute with water until colorless, then elute with 3 times the column volume of 30% ethanol, and then elute with 3 times the column volume of 70% ethanol, collect the 70% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0073] Example 15: Control group 3

[0074] On the basis of Example 2, 30% ethanol was added for elution, as follows:

[0075] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 80% ethanol in a volume-to-mass ratio of 12 times that of the medicinal materials, the extraction temperature is 80-90° C. Filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add 0.3 times the feed amount of 5% NaOH solution, add water to a total volume of 12 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, pass through HPD100 macroporous adsorption resin, elute with water until colorless, then elute with 3 times the column volume of 30% ethanol, and then elute with 3 times the column volume of 70% ethanol, collect the 70% ethanol eluate, concentrate the filtrate, dry, and crush to obtain.

[0076] Example 16: Control group 4

[0077] Weigh 200 g of dried ivy leaves, crush them into coarse powder, reflux extract twice with 80% ethanol at a volume mass ratio of 12 times that of the medicinal materials, the extraction temperature is 80-90°C, filter and combine the filtrate, recover the ethanol until there is no alcohol taste, add 0.3 times the feed amount of 5% sodium hydroxide solution, add water to a total volume of 12 times the feed amount, adjust the pH to 9.5-11, refrigerate and stand overnight, filter, adsorb with 5 times the feed amount of alkaline alumina, elute with 3 times the column volume of water, collect the water washing liquid, concentrate, dry, add an appropriate amount of methanol for ultrasonic extraction, filter, and concentrate and dry the filtrate.

[0078] The preparation process of Example 1 to Example 16 is shown in Table 1

[0079]

[0080]

[0081] Table 1 Preparation process

[0082] Example 17: Component Detection

[0083] Index test content: the content of saponin C, α-saponin, saponin B and saponin D in the extract.

[0084] Yield = extract weight / herbal material amount*100%.

[0085] Saponin transfer rate = extract weight * saponin content in the extract / (corresponding decoction piece weight * saponin content in the decoction piece) * 100%. 1. Experimental instruments and materials:

[0086] The production batch number of Hedera helix leaf slices is LX220501 (Zhejiang Kangenbei Chinese Medicine Co., Ltd.), the extracts are CCT 1-12, and the extracts of control groups 1-4 are prepared according to Examples 1-16.

[0087] 2. Detection method:

[0088] Chromatographic conditions and system suitability test: Chromatographic column ZORBAX Eclipse XDB-C18 Analytical 4.6*250mm5-Micron; acetonitrile as mobile phase A, 0.05% phosphoric acid solution as mobile phase B, gradient elution as specified in Table 2; column temperature 25°C; detection wavelength 205nm; injection volume 10μl, flow rate 1.0ml per minute.

[0089] Table 2 Gradient elution program

[0090]

[0091] Preparation of reference solution Take appropriate amount of Hederacoside C reference and α-hederacoside reference, weigh accurately, and add methanol to prepare a mixed solution containing 0.5 mg of Hederacoside C and 0.2 mg of α-hederacoside per 1 ml. Take appropriate amount of Hederacoside B and Hederacoside D reference, add methanol to prepare a solution containing 0.5 mg of Hederacoside B and Hederacoside D per 1 ml respectively.

[0092] 3. Experimental results

[0093] The results of the yield of extracts CCT 1-12, control groups 1 to 4, the content of saponin active ingredients in the extracts, and the saponin transfer rate are shown in Table 3. The results of the saponin transfer rate of extracts CCT-2, control groups 1 to 3 are shown in Table 4.

[0094] Table 3 Quality analysis results of ivy extracts prepared by different processes

[0095]

[0096] Table 4 Transfer rate of some extracts

[0097]

[0098]

[0099] By comparing Extract 2 and Control Group 1, it was found that the contents of saponin C and α-saponin in the extracts obtained under different elution conditions, different macroporous resin column fillers, and different alkalization conditions were similar, but the saponin transfer rate of Extract 2 was significantly higher than that of Control Group 1. Control Groups 2-4 were used to further study the effects of different preparation conditions on the content and transfer rate of components of ivy leaf extracts.

[0100] Control group 1 and control group 2 used different macroporous resin column fillers under the same alkalinization conditions and eluents. The content of the four saponin components in control group 2 was higher than that in control group 1. Extract 2 and control group 4 were eluted under the same conditions, and control group 4 used an alumina chromatography column, and the content of saponin components was significantly reduced. It can be seen that the enrichment of active ingredients is related to column chromatography fillers, and different chromatographic columns correspond to different active ingredients and different contents. From extracts 1 to 12, the yield and saponin content of the HPD100 macroporous resin column were the highest.

[0101] Control group 2 and control group 3 used the same chromatography column and eluent. Under different pH conditions, after increasing the amount of alkali solution, the contents of the four saponin components in the extract and the extract yield in control group 3 were all higher than those in control group 2, and the transfer rates of saponin C and α-saponin in control group 3 were relatively increased. It can be seen that increasing the amount of alkali solution increased the saponin transfer rate, yield and saponin content of the extract in a specific pH range. The reason is that increasing the amount of alkali solution can enhance the sedimentation of polysaccharides, proteins, etc. in the alcohol extract of ivy leaves, reduce the precipitation and encapsulation of effective ingredients, especially α-saponin, but excessive alkali concentration will destroy the 28-position ester bond of ivy saponin C and hydrolyze to form α-ivy saponin. When 0.3 times the alkali solution is used, the pH is adjusted in the range of 9.5-11, which not only increases the water precipitation effect of the alcohol extract, but also avoids the reduction of the transfer rate of ivy saponin C due to the formation of α-ivy saponin by alkaline hydrolysis.

[0102] Extracts 1 to 12 were obtained with a saponin content of 50 to 60% using HPD-100, polyamide and C18 columns. Except for Extract 2, the other extracts were all lower than the total saponin content obtained by eluting with DM130 + 30% + 70% ethanol in Control Group 1. In terms of yield, the yields of Extracts 1 to 3 were higher than that of Control Group 1, but the yields of Extracts 4 to 12 were lower than that of Control Group 1. From the conventional quality evaluation standards for extracts, the technical effects of high saponin content and yield are better, so Control Group 1 should be considered a better extraction method. However, the present invention tested the anti-inflammatory effect of the extract, and the results showed that the content of saponin components as the main active ingredients in the extract is not linearly correlated with the anti-inflammatory effect. Even if the saponin content is reduced, the anti-inflammatory effect obtained may be better, resulting in unexpected technical effects. See Example 18 for details.

[0103] Example 18: Evaluation of the inhibitory activity of ivy leaf extract on COX-2

[0104] 1. Experimental drugs and reagents

[0105] COX-2 inhibitor screening kit (Biyuntian Biotechnology), ivy extracts (CCT-1-12, control groups 1-4) (self-made by Zhejiang Kangenbei Chinese Medicine Co., Ltd.), DMSO (Diamond).

[0106] 2. Experimental Methods

[0107] According to the test requirements of the kit, the drug administration groups were set at three concentrations of 12.5μg / mL, 25μg / mL, and 50μg / mL, a positive drug group (celecoxib), and a DMSO group. According to the instructions of the kit, a 100% enzyme activity control group, a blank control group, a positive inhibitor group, and a sample group were set up. After adding the ivy leaf extract to be tested to the sample group well plate, mix it well and incubate it at 37°C for 10min. 5μL of COX-2Probe was added to each well of each group, and then 5μL of COX-2Substrate working solution was quickly added to each well, mixed well, and incubated at 37°C in the dark for 5 minutes before fluorescence measurement. The detection was carried out on a multifunctional microplate reader (Tecan), and the detection conditions were an excitation wavelength of 560nm and an emission wavelength of 590nm.

[0108] Inhibition rate (%) = (RFU 100％酶活性对照 -RFU 样品 ) / (RFU 100％酶活性对照 -RFU 空白对照 )×100%

[0109] 3. Experimental results

[0110] As shown in Table 5, the various ivy extracts have different inhibitory activities on COX-2, with half inhibition rates IC 50 See Table 6 below for details. According to the yield converted into the concentration of the decoction piece, the IC values ​​of extracts 1 to 12 can be obtained. 50 The values ​​were between 7.64 and 13.78 μg / mL, and the amount of the decoction pieces was converted to 113.52 to 328.10 μg / mL. The IC values ​​of control groups 1 to 4 50 The values ​​are between 35.92 and 249.9 μg / mL, and the converted medicinal material dosage is 520.58 to 6941.67 μg / mL.

[0111] Combined with the results of saponin content in Table 3, it can be seen that the content of the four saponins in control group 1 is 62.1%, which is slightly higher or similar to the saponin content of extracts 1 to 12, and the yield is also higher than that of extracts 4 to 12, but the corresponding IC after conversion of the decoction piece is 50 The value is 3 to 7 times that of extracts 1-12, and the corresponding anti-inflammatory activity is 1 / 7 to 1 / 3 of that of extracts 1 to 12. It can be seen that the saponin content corresponding to extracts 1 to 12 provided by the present invention is similar to or less than that of control group 1, but the anti-inflammatory effect is better. Saponin is the main active ingredient of ivy extract, but the technical scheme achieves better anti-inflammatory effect with similar or lower saponin content, which has unexpected technical effects. Therefore, the quality evaluation of the extract should not be limited to the yield and saponin content. A high saponin content does not necessarily mean a good anti-inflammatory effect, and there is no linear correlation between the saponin content and the efficacy.

[0112] The anti-inflammatory results of control group 1 and control group 2 showed that different chromatographic columns enriched different active ingredients, and the use of HPD100 macroporous resin column can better enrich the active ingredients than the DM130 macroporous resin column. Combining extracts 2, 5, 9 and control group 4, it was found that under the same alkaline solution pH value and eluent, the anti-inflammatory activity of the extracts obtained by HPD100 macroporous resin column, polyamide column, and C18 column was 30 to 58 times that of the extracts obtained by alumina column. It can be seen that HPD100, polyamide column, and C18 column can effectively enrich the anti-inflammatory components in the ivy leaf extract and increase the anti-inflammatory activity of the extract.

[0113] The anti-inflammatory effects of control group 2 and control group 3 showed that only a slight change in the amount of alkaline solution had little effect on the enrichment of anti-inflammatory components. However, in the purification process of crude extract of Hedera helix leaves, the selection of extraction solvent, alkaline solution, chromatographic column and eluent is an important part of the preparation method, which ultimately directly affects the pharmacological activity of Hedera helix leaf extract.

[0114] Comparing extract 2 with control group 3, the total amount of the four saponins in control group 3 was 74%, corresponding to IC 50 The value (converted to the amount of decoction pieces) was 531.92 μg / mL; the total amount of the four saponins in extract 2 was 62.5%, corresponding to IC 50 The value (converted to the amount of decoction pieces) was 120.11 μg / mL, which was 340% higher than the activity of control group 3. This shows that the crude extract of the helichrysum leaves of extract 2 was effectively enriched after separation by HPD100 column and elution with 70% ethanol; compared with the control group 3 in which high-purity saponins were obtained by gradient elution with 30% ethanol and 70% ethanol, extract 2 had a better anti-inflammatory effect. It can be seen that even if the same chromatographic column is used, the type and order of the eluent of the chromatographic column play an important role in the enrichment of anti-inflammatory components in the helichrysum leaf extract. The choice of eluent directly affects the type and content of the eluted components. Different elution methods enrich different anti-inflammatory components, and the elution method has an important influence on the efficacy of the extract.

[0115] Table 5 Inhibitory activity of ivy extracts on COX-2 enzyme

[0116]

[0117] Example 19: Pharmacodynamic effects of ivy leaf extract on lipopolysaccharide (LPS)-induced acute bronchitis model in mice 1. Experimental drugs and reagents

[0118] LPS (Sigma-Aldrich (Shanghai) Trading Co., Ltd.); Prospan oral solution (produced by Engelhard Arzneimittel, batch number: 22G080A); Ambroxol hydrochloride oral solution (produced by China Resources Sanjiu (Nanchang) Pharmaceutical Co., Ltd., batch number: 2112020J); Honey-refined Chuanbei loquat paste (produced by Kyoto Ninji General Factory Co., Ltd., batch number: Y12102104); CCT-2, CCT-5 and CCT-9 samples (prepared by Zhejiang Kangenbei Traditional Chinese Medicine Co., Ltd.).

[0119] 2. Animal grouping

[0120] After three days of adaptive feeding, SPF Balb / c mice were divided into 8 groups according to their body weight, with 10 mice in each group, as shown in Table 6 below.

[0121] Table 6 Animal grouping

[0122] Group Modeling treat Remark Blank control group Pure water Pure water N＝10 Model Group LPS Pure water N＝10 Dexamethasone group LPS 2 mg / kg dexamethasone N＝10 Honey-refined Chuanbei Loquat Paste LPS 5.8ml / kg Honey-refined Chuanbei loquat paste N＝10 Prospan Group LPS 3.2ml / kgProspan N＝10 CCT-2 group LPS 3.11mg / kgCCT-2 N＝10 CCT-5 group LPS 3.11mg / kgCCT-5 N＝10 CCT-9 Group LPS 3.11mg / kgCCT-9 N＝10

[0123] 3. Route and time of administration

[0124] Oral gavage, 0.2 ml / 10 g body weight for 3 consecutive days.

[0125] 4. Model replication

[0126] Healthy mice were taken and 6 mg / ml LPS saline solution was dripped into the nasal cavity at 10 mg / kg using a pipette for 3 consecutive days, and the mice were killed after 5 days. The drug was administered once a day, two days before modeling, and continued until modeling.

[0127] 5. Index detection

[0128] The animal serum was collected 1 hour after the last administration to detect the changes of inflammatory factor TNF-α.

[0129] 6. Statistical methods

[0130] All data were expressed as Mean±SD. The differences between each group and the model group were analyzed by t-test, with p<0.05 considered statistically significant.

[0131] 7. Experimental results

[0132] As shown in Table 7, the serum TNF-α of the mice in the model group increased at the end of administration, which was significantly different from that in the control group (P<0.01); compared with the model group, the drug groups could reduce the serum TNF-α at the end of administration, and 2mg / kg of dexamethasone acetate could reduce the level of TNF-α in serum, which was significantly different from that in the model group (P<0.01); 5.8ml / kg of Kyoto Nien Jiom Honey Refined Chuanbei Loquat had no difference compared with the model group, and 3.2ml / kg of Prospan could reduce the level of TNF-α in serum, which was significantly different from that in the model group (P<0.05); 3.11mg / kg of CCT-2, CCT-5, and CCT-9 had significant differences compared with the model group (P<0.05). The experimental results showed that CCT-2, CCT-5, and CCT-9 could effectively inhibit the growth of inflammatory factors in acute bronchitis in mice induced by LPS, had therapeutic effects, and had better anti-inflammatory effects than the Prospan group.

[0133] Table 7 Effect of ivy leaf extract on serum TNF-α in mice with acute bronchitis (n=10, Mean±SD)

[0134]

[0135] Note: **P<0.01 vs model group; *P<0.05 vs model group

[0136] Example 20: Study on the expectorant effect of ivy leaf extract on mouse expectorant model (phenol red method)

[0137] 1. Experimental drugs and reagents

[0138] Phenol red (GENERAL-REAGENT); ELISA reader (Thermo); drug sources are as shown in Example 19.

[0139] 2. Animal grouping

[0140] After three days of adaptation, 80 SPF ICR mice were randomly divided into 8 groups according to their body weight, with 10 mice in each group. The groups are shown in Table 8:

[0141] Table 8 Animal Grouping

[0142] Group Modeling treat Remark Blank control group Pure water Pure water N＝10 Model Group 5% phenol red Pure water N＝10 Ambroxol hydrochloride oral solution group 5% phenol red 2.57ml / kg ambroxol hydrochloride oral solution N＝10 Honey-refined Chuanbei Loquat Paste 5% phenol red 5.8ml / kg Honey-refined Chuanbei loquat paste N＝10 Prospan Group 5% phenol red 3.2ml / kgProspan N＝10 CCT-2 group 5% phenol red 3.11mg / kgCCT-2 N＝10 CCT-5 group 5% phenol red 3.11mg / kgCCT-5 N＝10 CCT-9 Group 5% phenol red 3.11mg / kgCCT-9 N＝10

[0143] 3. Route and time of administration

[0144] The drug was administered by oral gavage once a day for 3 consecutive days, with a dosing volume of 0.2 ml / 10 g body weight.

[0145] 4. Model replication

[0146] 30 minutes after the last administration of the mice, 0.1 ml / 10 g of 5% phenol red solution was injected intraperitoneally. The animals were killed 30 minutes later, the trachea was dissected and separated, and a flattened No. 7 needle was inserted into the trachea 0.3 cm, tied with silk thread, and 0.5 ml of 5% sodium bicarbonate solution was drawn with a 1 ml syringe. The respiratory tract was irrigated back and forth through the needle for 3 times, and the irrigating fluid was drawn out for the last time and injected into a test tube. The above operation was repeated 3 times, and about 1 ml of irrigating fluid was drawn, placed in a test tube, centrifuged at 1500 rpm for 5 minutes, and 150 μl of supernatant was drawn into a 96-well plate.

[0147] 5. Index detection

[0148] The OD value at 546 nm was measured with an enzyme-labeled instrument and adjusted to zero with 5% sodium bicarbonate solution.

[0149] 6. Statistical methods

[0150] All data were expressed as Mean±SD. The t-test was used to analyze the significance among groups. The differences between each group and the model group were tested, and p<0.05 was considered statistically significant.

[0151] 7. Experimental results

[0152] As shown in Table 9, 3.2ml / kg Prospan can increase the phenol red secretion of mouse trachea, which is significantly different from the model group (P<0.05); 2.57ml / kg ambroxol hydrochloride oral solution has a certain promoting effect on the phenol red secretion of mouse trachea, but there is no difference compared with the model group (P>0.05); 3.11mg / kg CCT-2, CCT-5 and CCT-9 can all increase the phenol red secretion of mouse trachea, which is significantly different from the model group (P<0.05); 5.8ml / kg honey refined Chuanbei loquat paste group has no obvious promoting effect. It can be seen that CCT-2, CCT-5 and CCT-9 can increase the phenol red secretion of mouse trachea, have a certain expectorant effect, and the expectorant effect is better than that of the Prospan group.

[0153] Table 9 Expectorant effect results of ivy leaf extract

[0154] Group dose OD value Blank control group / 0.159±0.054 Model Group / 0.180±0.042 Ambroxol hydrochloride oral solution group 2.57ml / kg 0.224±0.097 Honey-refined Chuanbei Loquat Paste 5.8ml / kg 0.172±0.040 Prospan Group 3.2ml / kg 0.224±0.044* CCT-2 group 3.11mg / kg 0.239±0.073* CCT-5 group 3.11mg / kg 0.229±0.038* CCT-9 Group 3.11mg / kg 0.231±0.040*

[0155] Note: *P<0.05vs model group

[0156] Example 21: Study on the antitussive effect of ivy leaf extract on mouse antitussive model (ammonia method)

[0157] 1. Experimental drugs and reagents

[0158] Ammonia water (Sinopharm Chemical Reagent Co., Ltd.); Dextromethorphan hydrobromide tablets (Guangzhou Baiyunshan Guanghua Pharmaceutical Co., Ltd.); the sources of the drugs are as shown in Example 19.

[0159] 2. Animal grouping

[0160] After three days of adaptive feeding, 70 SPF ICR female mice were divided into 7 groups according to their weight, with 10 mice in each group. The groups are shown in Table 10.

[0161] Table 10 Animal grouping

[0162] Group Modeling treat Remark Model Group 12% ammonia water Pure water N＝10 Dextromethorphan Hydrobromide Tablets 12% ammonia water 1.03 tablets / kg Dextromethorphan Hydrobromide Tablets N＝10 5.8ml / kg honey refined Chuanbei loquat paste group 12% ammonia water 5.8ml / kg Honey-refined Chuanbei loquat paste N＝10 3.2 ml / kg Prospan group 12% ammonia water 3.2ml / kgProspan N＝10 CCT-2 group 12% ammonia water 3.11mg / kgCCT-2 N＝10 CCT-5 group 12% ammonia water 3.11mg / kgCCT-5 N＝10 CCT-9 Group 12% ammonia water 3.11mg / kgCCT-9 N＝10

[0163] 3. Route and time of administration

[0164] Oral gavage, 0.2 ml / 10 g body weight for 3 consecutive days.

[0165] 4. Model replication

[0166] One hour after the last administration, the mice were placed in a YLS-8A multifunctional cough and wheeze inducing instrument and atomized with 12% ammonia for 10 seconds. The cough latency and the number of coughs within 2 minutes were observed and recorded.

[0167] 5. Index detection

[0168] Cough latency period, the total number of coughs within 2 minutes.

[0169] 6. Statistical methods

[0170] All data were expressed as Mean±SD. The differences between each group and the model group were analyzed by t-test, with p<0.05 considered statistically significant.

[0171] 7. Experimental results

[0172] As shown in Table 11, mice were given 3.2 ml / kg Prospan, 1.03 tablets / kg dextromethorphan hydrobromide, 5.8 ml / kg honey refined Chuanbei loquat paste, 3.11 mg / kg CCT-2, CCT-5 and CCT-9 for 3 consecutive days, which can prolong the cough latency of mice, and there is a significant difference compared with the model group (P<0.01). According to the Prospan oral solution instructions, the ratio of ivy dry extract to crude drug is 5-7.5:1, and the average 1 mg of ivy dry extract contains 6.25 mg of crude drug. The oral solution contains 7 mg of ivy dry extract per milliliter, and it can be inferred that 1 mL of oral solution contains 43.75 mg of ivy medicinal materials. The dosage of 3.2 ml / kg of Prospan oral solution corresponds to the amount of crude drug taken (calculated by mice) of 3.2*43.75=140 mg / kg. The yields of CCT-2, CCT-5 and CCT-9 were 9.0%, 3.9% and 4.3% respectively, and the corresponding crude drug amount was dose / yield, that is, the corresponding crude drug amount of CCT-2 was 3.11 / 9% = 34.44 mg / kg, the corresponding crude drug amount of CCT-5 was 3.11 / 3.9% = 79.74 mg / kg, and the corresponding crude drug amount of CCT-9 was 3.11 / 4.3% = 72.33 mg / kg. The experimental results showed that CCT-2, CCT-5 and CCT-9 could prolong the cough latency of mice, had a certain antitussive effect, and the corresponding crude drug amount was less than that of the Prospan group.

[0173] Table 11 Effect of ivy leaf extract on cough latency in mice (n=10, Mean±SD)

[0174] Group dose Cough latency period(s) Model Group / 20.9±5.9 Dextromethorphan Hydrobromide Tablets 1.03 pieces / kg 33.3±9.7** Honey-refined Chuanbei Loquat Paste 5.8ml / kg 40.8±10.5** Prospan Group 3.2ml / kg 38.6+8.3** CCT-2 group 3.11mg / kg 28.8±5.2** CCT-5 group 3.11mg / kg 29.2±7.1** CCT-9 Group 3.11mg / kg 30.0±8.2**

[0175] Note: **P<0.05vs model group

[0176] As shown in Table 12, mice were given 3.2ml / kg Prospan, 1.03 tablets / kg dextromethorphan hydrobromide, 5.8ml / kg honey refined Chuanbei loquat paste, 3.11mg / kg CCT-2, CCT-5 and CCT-9 for 3 consecutive days, which can reduce the number of coughs in mice, and there is a significant difference compared with the model group (P<0.01). The experimental results show that CCT-2, CCT-5 and CCT-9 can reduce the number of coughs in mice, have a certain antitussive effect, and the corresponding raw drug dosage is less than that of the Prospan group.

[0177] Table 12 Effect of ivy leaf extract on the total number of coughs in mice within 2 minutes (n=10, Mean±SD)

[0178] Group dose frequency Model Group / 27.9±4.5 Dextromethorphan Hydrobromide Tablets 1.03 pieces / kg 17.1±6.2** Honey-refined Chuanbei Loquat Paste 5.8ml / kg 17.9±5.3** Prospan Group 3.2ml / kg 17.1±3.3** CCT-2 group 3.11mg / kg 20.6±3.2** CCT-5 group 3.11mg / kg 20.7±4.6** CCT-9 Group 3.11mg / kg 21.3±3.2**

[0179] Note: **P<0.05vs model group

Claims

1. A method for preparing an ivy extract, characterized in that: The method comprises the following steps: extracting ivy leaves with 75% to 85% ethanol, concentrating and adjusting the pH to 9.5 to 11, standing, filtering and passing the obtained crude extract through a chromatography column, eluting with 60% to 80% ethanol, and obtaining the ivy extract; the chromatography column is one of an HPD100 macroporous resin column, a polyamide column or a C18 column.

2. The method according to claim 1, characterized in that When the chromatography column is a polyamide column, the pH of the crude extract is adjusted to neutral and then passed through the chromatography column; When the chromatography column is a C18 column, the crude extract is passed through the chromatography column, first eluted with 30% ethanol, then eluted with 60%-80% ethanol, and the eluate eluted with 60%-80% ethanol is collected.

3. The method according to claim 1, characterized in that The method comprises the following steps: (1) Take ivy leaves, crush them, extract them with 75% to 85% ethanol under reflux, and filter and combine the filtrate; (2) The filtrate is concentrated until there is no alcohol taste, the pH is adjusted to 9.5 to 11, refrigerated and allowed to stand for 8 to 15 hours, and filtered to obtain a crude extract; (3) The crude extract is passed through a chromatography column, eluted with water until colorless, and then eluted with 3 to 5 times the column volume of 60% to 80% ethanol. The ethanol eluate is collected, concentrated and dried to obtain the ivy extract; the chromatography column is one of an HPD100 macroporous resin column, a polyamide column or a C18 column.

4. The method according to claim 3, characterized in that The chromatography column is a HPD100 macroporous resin column.

5. The method according to claim 3, characterized in that In the step (2), an alkaline solution is used to adjust the pH value, and the alkaline solution is a sodium hydroxide solution or a potassium hydroxide solution.

6. The method according to claim 5, characterized in that In the step (2), the concentration of the sodium hydroxide solution or potassium hydroxide solution is 3-8%, and the volume of the alkaline solution is 0.2-0.4 times the amount of ivy leaves added.

7. The method according to claim 5, characterized in that In the step (2), the mass of the sodium hydroxide or potassium hydroxide in the alkaline solution is 1-2% of the amount of ivy leaves added.

8. The method according to claim 5, characterized in that In the step (2), after the pH value of the alkaline solution is adjusted, water is added to a total volume that is 5 to 12 times the amount of ivy leaves added.

9. The ivy extract obtained by the preparation method according to any one of claims 1 to 8.

10. The ivy extract according to claim 9, characterized in that The ivy extract comprises α-hedera saponin, hedera saponin C, hedera saponin B and hedera saponin D, and the total mass fraction of α-hedera saponin, hedera saponin C, hedera saponin B and hedera saponin D is greater than 50%.

11. The ivy extract according to claim 10, characterized in that In the ivy extract, the mass content of α-hedera saponin is greater than 10%, the mass content of hedera saponin C is greater than 30%, the mass content of hedera saponin B is greater than 2%, and the mass content of hedera saponin D is greater than 1%.

12. Use of the ivy extract obtained by the preparation method according to any one of claims 1 to 8 or the ivy extract according to any one of claims 10 to 11 in the preparation of a drug for treating or preventing acute bronchitis.

13. Use of the ivy extract obtained by the preparation method according to any one of claims 1 to 8 or the ivy extract according to any one of claims 10 to 11 in the preparation of expectorant and antitussive drugs.

14. A drug for treating or preventing acute bronchitis, comprising the ivy extract prepared by the preparation method according to any one of claims 1 to 8 or the ivy extract according to any one of claims 10 to 11.

15. An expectorant and antitussive drug, comprising the ivy extract obtained by the preparation method according to any one of claims 1 to 8 or the ivy extract according to any one of claims 10 to 11.

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