Composition of salvianolic acid extract and tanshinone extract as well as preparation method, application and product thereof

Abstract

The invention provides a composition of a salvianolic acid extract and a tanshinone extract as well as a preparation method, application and a product of the composition, and belongs to the field of medicines. The preparation method of the composition of the salvianolic acid extract and the tanshinone extract comprises steps 1)-4). The composition with carbonic anhydrase inhibitory activity is obtained by extracting the salvianolic acid extract and the tanshinone extract from the radix salviae miltiorrhizae and mixing the salvianolic acid extract and the tanshinone extract, the composition can achieve synergistic interaction, and reference is provided for research and development and component compatibility of medicines mainly comprising the radix salviae miltiorrhizae. The composition provided by the invention can be used for reducing intraocular pressure, inducing diuresis, correcting metabolic acidosis, increasing vascular oxygen flux, improving pulmonary ventilatory capacity, expanding cerebral vessels, inducing tumor cell apoptosis and inhibiting osteoclast bone resorption; the traditional Chinese medicine composition can be used for treating or preventing diseases with intraocular hypertension as symptoms, cardiac edema or acute altitude sickness, cardiovascular and cerebrovascular diseases and bone metabolism diseases caused by osteoporosis.

Description

Technical Field

[0001] The present invention belongs to the field of medicine and relates to a composition of salvianolic acid and tanshinone, and specifically relates to a composition of salvianolic acid extract and tanshinone extract, and a preparation method, application and product thereof. Background Art

[0002] Salvia miltiorrhiza is a traditional Chinese medicine with the effects of promoting blood circulation and removing blood stasis, relieving pain, cooling blood and eliminating carbuncle, anti-inflammatory, anti-oxidation, protecting myocardial cells, protecting blood vessels, etc. It is often used to treat cardiovascular diseases such as atherosclerosis, coronary heart disease, and hypertension. The components of Salvia miltiorrhiza can be divided into water-soluble phenolic acids and fat-soluble diterpenoid tanshinones. Among them, the phenolic acid components of Salvia miltiorrhiza are mainly composed of components such as salvianolic acid B, salvianolic acid A, rosmarinic acid, and lithospermic acid, while the tanshinone components mainly include tanshinone IIA, cryptotanshinone, dihydrotanshinone I, and tanshinone I.

[0003] Tanshinone compounds are the main active substances for the treatment of cardiovascular and cerebrovascular diseases. They are mainly distributed in the periderm cork layer of the roots and rhizomes of Salvia miltiorrhiza, followed by the phloem. The color distribution of the periderm cork layer cells is positively correlated with their content, that is, the stronger the red intensity of the cork layer cells, the higher the content of tanshinone components. However, there is no direct correlation between the color of xylem cells and the distribution of tanshinone components. The xylem is the main distribution site of salvianolic acid compounds, followed by the phloem. The content of salvianolic acid in the cork layer is very low. Studies in recent years have shown that the aboveground stems, leaves and inflorescences of Salvia miltiorrhiza also have pharmacological effects similar to those of roots and rhizomes. The content of salvianolic acid compounds is significantly higher than that of underground roots and rhizomes during the vigorous growth period.

[0004] The conference paper (Liu Qingping. Clinical observation on the treatment of patients with dilated cardiomyopathy and chronic heart failure with Qili Qiangxin Capsule [C]. Yichun Second People's Hospital, Jiangxi Province, 2024) disclosed that tanshinone and tanshinol acid components contained in Salvia miltiorrhiza have significant therapeutic effects on cardiovascular and cerebrovascular diseases such as chronic heart failure, coronary atherosclerosis, and angina pectoris. The reason is that tanshinone and tanshinol acid have cardiovascular protective effects such as reducing the inflammatory response of myocardial cells, inhibiting cell adhesion, anti-inflammatory and antioxidant, and protecting cardiovascular endothelial cells.

[0005] The invention patent with publication number CN115192585A discloses a composition comprising salvianolic acid A, dihydrotanshinone I and cryptotanshinone and its use. In the composition comprising salvianolic acid A, dihydrotanshinone I and cryptotanshinone, the molar ratio of salvianolic acid A, dihydrotanshinone I and cryptotanshinone is 14:14:14. The composition can significantly reduce the serum transaminase level of mice with liver fibrosis model, reduce liver tissue inflammation and collagen deposition, inhibit the secretion of proinflammatory cytokines by proinflammatory macrophages, and inhibit the activation of proinflammatory macrophage inflammasomes. It is confirmed that the molar ratios of salvianolic acid A, dihydrotanshinone I and cryptotanshinone are optimal at 1:3:4 and 3:4:2.

[0006] The invention patent with publication number CN101863957A discloses a method for synchronously and efficiently extracting tanshinone ⅡA and salvianolic acid B from salvia miltiorrhiza. The salvia miltiorrhiza is crushed and extracted with a hydrophilic solvent under low temperature conditions. The pressure is continuously changed during the extraction process, and the extract is concentrated to obtain an extract rich in tanshinone ⅡA and salvianolic acid B. The invention uses a small amount of organic solvent, takes a short time, and has a high extraction rate. The extraction process achieves the purpose of efficient extraction by continuously changing the pressure. The extraction and concentration process is processed under low temperature conditions to ensure the extraction efficiency and not destroy the effective ingredients. A pressure pump is used to circulate the solvent during the extraction process to improve the extraction efficiency.

[0007] Carbonic anhydrase (CA) is a zinc-containing metalloenzyme that catalyzes the 2 CA is widely distributed in human tissues and participates in respiration, CO 2 and HCO 3 - It is an important link in transmembrane transport, acid-base balance, osmotic pressure regulation, calcification, signal transduction, tumorigenesis, etc.

[0008] The development of carbonic anhydrase inhibitors (CAI) has a long history. CAI, led by sulfonamide drugs such as acetazolamide, reduces H by inhibiting CA activity. + -Na + Exchange, HCO 3 - Transport and other physiological processes, thereby achieving the effects of improving metabolic acidosis, lowering intraocular pressure, and diuresis. It is often used in the treatment of glaucoma, cardiogenic edema, and acute mountain sickness. However, acetazolamide can cause numbness and tingling in the limbs, fatigue, weight loss, drowsiness, depression, lethargy, metallic taste, nausea, loss of appetite, indigestion, diarrhea, polyuria, nocturia, kidney and urinary tract knots, temporary myopia, sulfonamide rash, and exfoliative dermatitis. Therefore, it is necessary to further develop carbonic anhydrase inhibitors with low side effects and good activity.

[0009] The journal literature (Zhu Fangshi et al. Research progress and review on animal models of atrophic gastritis using traditional Chinese medicine [J]. Journal of Shanghai University of Traditional Chinese Medicine, 2003, (03): 61-64) disclosed that Professor Tung Chee-hwa's "Ganping Yangwei" prescription (Pseudostellaria heterophylla, roasted lily bulb, citron peel, etc.) and "Wei Le" prescription (Salvia miltiorrhiza, Panax notoginseng powder, Hedyotis diffusa, Scutellaria barbata, etc.) were used for enzyme histochemistry and electron microscopy observation on experimental CAG. The results showed that the aqueous solution of the two prescriptions can enhance the activities of succinate dehydrogenase (SDH) and carbonic anhydrase (CA) and reduce the activity of LDH.

[0010] Both salvianolic acid components and tanshinone components have cardiovascular protection, nerve cell protection, anti-inflammatory and antioxidant effects, but the above technologies do not disclose that salvianolic acid components and tanshinone components have inhibitory activity on carbonic anhydrase, so further research is necessary. Summary of the invention

[0011] The present invention aims at the problems existing in the prior art and provides a composition of salvianolic acid extract and tanshinone extract and a preparation method, application and product thereof. The present invention obtains salvianolic acid extract and tanshinone extract by extracting from salvia miltiorrhiza, mixes the salvianolic acid extract and tanshinone extract to obtain a composition with carbonic anhydrase inhibitory activity, and further adjusts the dosage ratio of salvianolic acid extract and tanshinone extract in the composition to synergistically enhance the carbonic anhydrase inhibitory activity.

[0012] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0013] In a first aspect, the present invention provides a method for preparing a composition of salvianolic acid extract and tanshinone extract, comprising the following steps:

[0014] 1) adding ethanol aqueous solution to salvia miltiorrhiza for percolation extraction, filtering, and collecting salvia miltiorrhiza extract;

[0015] 2) removing ethanol from the Danshen extract obtained in step 1), adjusting the pH to acidic, standing, separating the filtrate and the filter residue, washing and drying the filter residue to obtain a tanshinone extract;

[0016] 3) The filtrate obtained in step 2) is sequentially subjected to resin adsorption, impurity removal and elution, and the eluate is collected, concentrated and dried to obtain a salvianolic acid extract;

[0017] 4) The tanshinone extract obtained in step 2) and the salvianolic acid extract obtained in step 3) are mixed to obtain a composition of salvianolic acid extract and tanshinone extract.

[0018] Specifically, the volume fraction of the ethanol aqueous solution in step 1) is 10-90%. In one embodiment, the step 1) further includes swelling before the percolation extraction, and step 1) specifically comprises: adding ethanol aqueous solution to the salvia miltiorrhiza to perform swelling, percolation extraction, filtering, and collecting the salvia miltiorrhiza extract.

[0019] The percolation extraction is a method of placing the crushed Salvia miltiorrhiza in a percolation cylinder, continuously adding solvent from the top, and leaching the Salvia miltiorrhiza components during the process of the solvent permeating through the Salvia miltiorrhiza layer and flowing downward. It is a dynamic leaching method with high solvent utilization, complete leaching of effective components, and the leachate can be directly collected. The percolation extraction method can be a single percolation method, a heavy percolation method, a pressurized percolation method, or a countercurrent percolation method. In one embodiment, after the Salvia miltiorrhiza is crushed, an 80% volume fraction ethanol aqueous solution is added to soak and swell for 3 hours, and the ratio of Salvia miltiorrhiza to the ethanol aqueous solution is 1g:1-1.2mL, and then percolation extraction is performed; the percolation extraction includes: using 1080mL of 80% volume fraction ethanol aqueous solution to percolate for 5 hours, and the ratio of Salvia miltiorrhiza to the ethanol aqueous solution in the percolation extraction is 1g:9-9.2mL, and filtering and collecting the Salvia miltiorrhiza extract. The Salvia miltiorrhiza extract is subjected to rotary evaporation to remove ethanol, and the Salvia miltiorrhiza extract is obtained after freeze drying. In one embodiment, after crushing Danshen, 118g of powder was weighed, and 120mL of 80% by volume ethanol aqueous solution was added to soak and swell for 3h, and then 1080mL of 80% by volume ethanol water was used for percolation for 5h, and Danshen extract was collected by filtration. The Danshen extract was subjected to rotary evaporation to remove ethanol, and Danshen extract was obtained after freeze drying.

[0020] In the step 2), hydrochloric acid is used to adjust the pH to 2.0-4.0, preferably 3.0, and the standing time is 12-48 hours, preferably overnight.

[0021] In the step 3), the filtrate is sequentially subjected to resin adsorption, impurity removal and elution, specifically as follows: the filtrate is loaded onto an adsorption resin for adsorption, the adsorption resin is preferably D101 macroporous adsorption resin, and impurities are removed using pure water and 20-40% by volume ethanol aqueous solution in sequence, the column volume of the pure water is 1-10, preferably 10; the volume fraction of the ethanol aqueous solution is preferably 30%, and the column volume is 1-10, preferably 7; then 4-8 column volumes of 40-60% by volume ethanol aqueous solution are used for elution, and the column volume of the elution is preferably 6.

[0022] In the composition of step 4), the mass ratio of the salvianolic acid extract to the tanshinone extract is 1-100:1, preferably 2.5-30:1, more preferably 5-10:1. In one embodiment, the mass ratio of the salvianolic acid extract to the tanshinone extract is 9-10:1.

[0023] In a second aspect, the present invention provides a composition of salvianolic acid extract and tanshinone extract, which is prepared by the preparation method described in the above technical solution.

[0024] In a third aspect, the present invention provides a composition prepared by the preparation method described in the above technical solution or use of the composition described in the above technical solution in a drug for inhibiting carbonic anhydrase activity.

[0025] The medicine includes medicines for treating or preventing diseases with high intraocular pressure as symptoms, treating or preventing cardiogenic edema, and treating or preventing altitude sickness. The present invention has no particular restrictions on the altitude sickness, and those familiar to those skilled in the art are sufficient. It can be a disease with altitude sickness as symptoms, and the altitude sickness includes headache, dizziness, nausea, drowsiness, shortness of breath, palpitations, vomiting, fatigue, insomnia, numbness of hands and feet, or arrhythmia. In one embodiment, the altitude sickness is chronic altitude sickness or acute altitude sickness caused by increased altitude. The present invention has no particular restrictions on the disease with high intraocular pressure as symptoms, and those familiar to those skilled in the art are sufficient. In one embodiment, the medicine includes medicines for treating or preventing diseases with high intraocular pressure as symptoms, and the diseases with high intraocular pressure as symptoms are glaucoma, cataracts, or macular edema.

[0026] In a fourth aspect, the present invention provides a medicine, including a composition prepared by the preparation method described in the above technical scheme or a composition described in the above technical scheme.

[0027] The drug also includes at least one pharmaceutical excipient, which varies depending on the route of administration and the characteristics of action, and is a conventional pharmaceutical excipient in the pharmaceutical field, including but not limited to fillers, binders, wetting agents, absorption promoters, surfactants, flavoring agents or sweeteners. The filler can be starch, sucrose or microcrystalline cellulose; the binder can be starch slurry, hydroxypropyl cellulose, gelatin or polyethylene glycol; the wetting agent can be magnesium stearate, micropowder silica gel or polyethylene glycol; the absorption promoter can be polysorbate or lecithin; the surfactant can be poloxamer, fatty acid sorbitan or polysorbate.

[0028] The dosage form of the drug is tablet, pill, powder, capsule, solution, suspension, emulsion, aerosol or suppository. The tablet can be a dispersible tablet; the capsule can be a soft capsule, a hard capsule or a microcapsule; the solution can be an elixir, a syrup or an injection; the emulsion can be an oral emulsion, an intravenous emulsion or a liniment, etc. The drug can be made into a conventional, immediate release or sustained release preparation.

[0029] The drug is administered orally, by nasal inhalation, by skin, by cavity or by injection, and the injection includes intramuscular injection, subcutaneous injection or intravenous injection.

[0030] Compared with the prior art, the present invention has the following beneficial effects: the present invention obtains salvianolic acid extract and tanshinone extract by extracting from salvia miltiorrhiza, mixes the salvianolic acid extract and tanshinone extract to obtain a composition with carbonic anhydrase inhibitory activity, and further adjusts the dosage ratio of the salvianolic acid extract and the tanshinone extract in the composition to synergistically enhance the carbonic anhydrase inhibitory activity, thereby providing a reference for drug development and component composition based on salvia miltiorrhiza. The composition of the salvianolic acid extract and the tanshinone extract can be used to reduce intraocular pressure, promote diuresis, correct metabolic acidosis, increase vascular oxygen flux, increase pulmonary ventilation, dilate cerebral blood vessels, induce tumor cell apoptosis, and inhibit osteoclast bone resorption, thereby developing drugs with better efficacy for treating or preventing diseases with high intraocular pressure as symptoms, cardiogenic edema or acute mountain sickness, cardiovascular and cerebrovascular diseases, and bone metabolic diseases caused by osteoporosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The diagrams are the result of molecular docking simulation of salvianolic acid A and tanshinone IIA with CA, wherein A is the result of molecular docking of salvianolic acid A with CA, and B is the result of molecular docking of tanshinone IIA with CA;

[0032] Figure 2 is a graph showing the CA inhibition rate of a combination of salvianolic acid extract and tanshinone extract in a mass ratio of 10:1;

[0033] Figure 3 is a graph showing the CA inhibition rate of a combination of salvianolic acid extract and tanshinone extract at a mass ratio of 5:1;

[0034] Figure 4 is a graph showing the CA inhibition rate of a composition of salvianolic acid extract and tanshinone extract at a mass ratio of 2.5:1;

[0035] Figure 5 This is a graph showing the CA inhibition rate of a combination of salvianolic acid extract and tanshinone extract in a mass ratio of 1:1. DETAILED DESCRIPTION

[0036] It is worth noting that, unless otherwise specified, the raw materials used in the present invention are all common commercially available products, and their sources are not specifically limited.

[0037] As an example:

[0038] Buffer: 3-morpholinepropanesulfonic acid (MOPS).

[0039] Example 1

[0040] Salvianolic acid A is the main component of the salvianolic acid component, and tanshinone IIA is the main component of the tanshinone component. Therefore, salvianolic acid A and tanshinone IIA were used to carry out molecular docking with CA respectively. The molecular docking process is as follows:

[0041] A1. Acquisition of structural files: Small molecule structure files (tanshinone A and tanshinone IIA) were obtained through the Pubchem database (https: / / pubchem.ncbi.nlm.nih.gov / ), and CA protein structure files were obtained through the RCSB PDB database (https: / / www.rcsb.org / ).

[0042] A2. Molecular docking: Pymol software was used to remove water molecules and hydrogenate the structure file. The processed small molecule and CA structure file were imported into the AutoDock program for docking. The docking was performed by the Genetic Algorithm algorithm for 50 operations, and the result with the lowest binding energy was derived.

[0043] A3. Result visualization: Import the result with the lowest binding energy into the Protein-Ligand InteractionProfiler website (https: / / plip-tool.biotec.tu-dresden.de / ) for visualization to obtain detailed information on the interaction force. Finally, import the docking results into the Pymol software for visualization, and color the interaction force and molecular structure.

[0044] The molecular docking results are as follows Figure 1 As shown, Figure 1 The figure shows the result of molecular docking simulation of salvianolic acid A and tanshinone IIA with CA, where A is the molecular docking result of salvianolic acid A with CA, and B is the molecular docking result of tanshinone IIA with CA. Molecular docking revealed that salvianolic acid components and tanshinone components have different mechanisms of inhibiting CA. The salvianolic acid components (salvianolic acid A) are fixed to the active pocket of CA through hydrogen bonds and coordinate with zinc ions to inhibit CA activity, while the tanshinone components (tanshinone IIA) are fixed to the active pocket through hydrophobic forces to inhibit activity.

[0045] Example 2

[0046] Based on the results of Example 1, the carbonic anhydrase inhibitory activity of the combination of salvianolic acid extract and tanshinone extract was further determined.

[0047] The active composition is composed of salvianolic acid extract and tanshinone extract, and the combined effect thereof is evaluated using the Bliss independence model, specifically comprising the following steps:

[0048] B1. Preparation of sample solution: After crushing Danshen, weigh 118g of powder, add 120mL of 80% volume fraction ethanol aqueous solution to soak and swell for 3h, then use a percolation cylinder, use 1080mL of 80% volume fraction ethanol water to percolate for 5h, and filter and collect Danshen extract. The Danshen extract is evaporated to remove ethanol, and hydrochloric acid is added to adjust the pH to 3.0. Let it stand overnight, collect the filter residue after filtration, wash it repeatedly with pure water, and freeze-dry to obtain the tanshinone extract. The filtrate is loaded on D101 macroporous adsorption resin, and 7 column volumes of pure water and 7 column volumes of 30% volume fraction ethanol aqueous solution are used for impurity removal, and 7 column volumes of 50% volume fraction ethanol are used for elution, and the eluate is collected, concentrated by rotary evaporation, and freeze-dried to obtain the tanshinone acid component. The salvianolic acid extract is dissolved in a buffer solution to prepare a 0.1-5000 μg / mL solution; the tanshinone extract is dissolved in dimethyl sulfoxide (DMSO) to prepare a 50 mg / mL stock solution, and then a buffer solution is added to dilute it to a 0.1-5000 μg / mL solution; the prepared tanshinone solution and the salvianolic acid solution are mixed to prepare a composition composed of salvianolic acid extract and tanshinone extract in different mass ratios, wherein the mass ratios of salvianolic acid extract and tanshinone extract in the composition solution are 10:1, 5:1, 2.5:1, and 1:1, respectively.

[0049] B2. Verification of synergistic effect: The compositions of different mass ratios obtained in step B1 (100 μg: 10 μg, 50 μg: 10 μg, 100 μg: 40 μg, 40 μg: 40 μg), salvianolic acid extract solutions with concentrations of 40 μg / mL, 50 μg / mL, and 100 μg / mL, and tanshinone extract solutions with concentrations of 10 μg / mL, 20 μg / mL, and 40 μg / mL were used as test samples. 40 μL of the test samples were added to a 96-well plate, mixed with 50 μL of p-nitrophenyl acetate (PNPA, 6 mM) solution, and then 10 μL of carbonic anhydrase solution (100 nM) was added to start the reaction. The well plate was placed in a constant temperature mixer and incubated at 37°C and 600 r / min for 20 min to obtain the samples to be tested. The samples to be tested were detected using an ELISA instrument at 405 nm and the absorbance value of each well was recorded.

[0050] The difference between control sample 1 and the test sample is that control sample 1 uses buffer to replace the carbonic anhydrase solution. The specific preparation method of control sample 1 is: 40 μL of the test sample is added to a 96-well plate, mixed with 50 μL of p-nitrophenyl acetate (6 mM) solution, and then 10 μL of buffer is added to react to obtain control sample 1.

[0051] The difference between control sample 2 and the test sample is that control sample 2 uses buffer to replace the test sample. The preparation method of control sample 2 is as follows: 40 μL of buffer is added to a 96-well plate, mixed with 50 μL of p-nitrophenyl acetate (6 mM) solution, and then 10 μL of carbonic anhydrase solution (100 nM) is added to start the reaction to obtain control sample 2.

[0052] The difference between control sample 3 and the test sample is that control sample 3 uses buffer to replace the carbonic anhydrase solution and the test sample. The preparation method of control sample 3 is as follows: 40 μL of buffer is added to a 96-well plate, mixed with 50 μL of p-nitrophenyl acetate (6 mM) solution, and then 10 μL of buffer is added to react to obtain control sample 3.

[0053] Calculate the inhibition rate of the sample to be tested according to the inhibition rate calculation formula ①.

[0054] Inhibition rate % = [1-(A 1 -A 2 ) / (A 3 -A 4 )]×100%①

[0055] Among them, A 1 Represents the absorbance of the well containing the sample to be tested; A 2 represents the absorbance of the well containing control sample 1; A 3 represents the absorbance of the well containing control sample 2; A 4 The absorbance of the well representing control sample 3.

[0056] B3. Verification of synergistic effect: Calculate the expected additive reaction and combination index (CI) by using formulas ② and ③:

[0057] Expected Additive Effect = E A + E B -E A E B ②

[0058] CI=(E A +E B -E A E B ) / E AB ③

[0059] Among them, E A 、E B represents the inhibition rate of a single compound, E AB represents the inhibition rate of the composition. If CI is less than 1, and E AB This is greater than the expected additive effect, demonstrating the existence of a synergistic effect between the two compounds.

[0060] Synergy, also known as synergistic effect, refers to the situation where when two or more different compounds, components, or extracts are mixed together, the effect they can exert is greater than the sum of the effects of each component alone. Synergistic effect can be measured using the Blissindependence model, which evaluates whether two components have synergistic effects by calculating the expected additive reaction. If the effect produced by the mixture is greater than the expected additive effect, it indicates synergy; otherwise, it indicates antagonism.

[0061] The results are as follows Figure 2-5 As shown, Figure 2-5 The CA inhibition rate diagrams of the composition of salvianolic acid extract and tanshinone extract with mass ratios of 10:1, 5:1, 2.5:1, and 1:1 are shown respectively, where the dotted line represents the value of the expected additive effect. The results show that the inhibition rate of the composition is higher than the expected additive effect when the concentration ratio is 10:1 and 5:1, and the CI is less than 1. This indicates that the composition of salvianolic acid extract and tanshinone extract has a synergistic effect at a concentration ratio of 10:1 and 5:1 and can inhibit CA activity. Salvianolic acid extract and tanshinone extract can achieve the purpose of increasing drug efficacy, reducing side effects, and prolonging drug action time by exerting synergistic effects. The composition of salvianolic acid extract and tanshinone extract can be used to reduce intraocular pressure, promote diuresis, correct metabolic acidosis, increase vascular oxygen flux, improve pulmonary ventilation, dilate cerebral blood vessels, induce tumor cell apoptosis, inhibit osteoclastic bone resorption, and can also be used to treat and / or prevent cardiovascular and cerebrovascular diseases, eye diseases with high intraocular pressure as symptoms, altitude sickness, and bone metabolic diseases caused by osteoporosis.

[0062] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing a composition of salvianolic acid extract and tanshinone extract, characterized in that: The following steps are involved: 1) adding ethanol aqueous solution to salvia miltiorrhiza for percolation extraction, filtering, and collecting salvia miltiorrhiza extract; 2) removing ethanol from the Danshen extract obtained in step 1), adjusting the pH to acidic, standing, separating the filtrate and the filter residue, washing and drying the filter residue to obtain a tanshinone extract; 3) The filtrate obtained in step 2) is sequentially subjected to resin adsorption, impurity removal and elution, and the eluate is collected, concentrated and dried to obtain a salvianolic acid extract; 4) The tanshinone extract obtained in step 2) and the salvianolic acid extract obtained in step 3) are mixed to obtain a composition of salvianolic acid extract and tanshinone extract.

2. The preparation method according to claim 1, characterized in that: In the step 1), the volume fraction of the ethanol aqueous solution is 10-90%; in the step 2), hydrochloric acid is used to adjust the pH, and the pH is 2.0-4.

0.

3. The preparation method according to claim 1, characterized in that: In the step 3), the filtrate is sequentially subjected to resin adsorption, impurity removal and elution, specifically: the filtrate is loaded onto an adsorption resin for adsorption, 1-10 column volumes of pure water and 1-10 column volumes of 20-40% volume fraction ethanol are sequentially used for impurity removal, and then 4-8 column volumes of 40-60% volume fraction ethanol are used for elution.

4. The preparation method according to claim 1, characterized in that: In the composition of step 4), the mass ratio of salvianolic acid extract to tanshinone extract is 1-100:

1.

5. The preparation method according to claim 4, characterized in that: The mass ratio of the salvianolic acid extract to the tanshinone extract is 2.5-30:

1.

6. The preparation method according to claim 5, characterized in that: The mass ratio of the salvianolic acid extract to the tanshinone extract is 5-10:

1.

7. A composition of salvianolic acid extract and tanshinone extract, characterized in that: The compound is prepared by the preparation method according to any one of claims 1 to 6.

8. Use of the composition prepared by the preparation method according to any one of claims 1 to 6 or the composition according to claim 7 in a drug for inhibiting carbonic anhydrase activity.

9. The use according to claim 8, characterized in that: The medicine includes medicine for treating or preventing diseases with high intraocular pressure as a symptom, treating or preventing cardiogenic edema, and treating or preventing altitude sickness; the disease with high intraocular pressure as a symptom is glaucoma, cataract or macular edema.

10. A drug, characterized in that The invention comprises a composition prepared by the preparation method according to any one of claims 1 to 6 or a composition according to claim 7.

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