Pharmaceutical composition and application thereof

Abstract

The invention relates to a pharmaceutical composition containing pine needle and kudzuvine root extracts and application thereof, and the pharmaceutical composition can more stably and effectively exert curative effect by controlling the content of the pine needle and kudzuvine root extracts and the proportion of each component in the extracts.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a pharmaceutical composition and application thereof. Background Art

[0003] Hyperlipidemia refers to a disease in which the concentration of one or more components of plasma lipids exceeds the upper limit of normal [abnormal increase in cholesterol (TC) and triglycerides (TG) alone or in combination]. From the perspective of the main clinical symptoms, it can be summarized into the categories of TCM diseases such as "vertigo", "chest pain", and "stroke". The liver (gallbladder), kidney, and spleen (stomach) are considered to be the main sites of lesions; kidney essence deficiency and spleen dysfunction are the main internal causes; phlegm turbidity coagulates and is injected into the blood vessels, which is the key pathogenesis; phlegm and blood stasis are intertwined, deposited in the blood, and the loss of flexibility in the blood vessels is the inevitable outcome of its development into cardiovascular and cerebrovascular diseases. Due to the continuous improvement of living standards, the incidence of hyperlipidemia has gradually increased and tends to develop in the middle-aged population. Hyperlipidemia is an important risk factor for cardiovascular and cerebrovascular diseases and the primary risk factor for atherosclerosis (AS). Cardiovascular and cerebrovascular diseases caused by AS are one of the main diseases that currently endanger human health and life. In addition, hyperlipidemia can also lead to presbycusis, fatty liver, and aggravation of progressive nephrotic syndrome, etc. Therefore, hyperlipidemia has become one of the hot spots in metabolic disease research in the past decade. Among them, the seven-year coronary heart disease prevention and treatment study conducted by the American Heart, Lung and Blood Institute confirmed that the use of lipid-lowering drugs can reduce the mortality rate of heart disease by 24% and the rate of non-fatal heart attacks by 19%. At the same time, lipid-lowering drugs can also delay or alleviate the development of AS lesions and promote their regression.

[0004] At present, there have been a certain number of studies on traditional Chinese medicine for the improvement of hypertension and hyperlipidemia. However, in the current research on traditional Chinese medicine, although a single component has certain efficacy, the therapeutic effect is limited, and multiple components can usually play a synergistic role between the efficacy, becoming the mainstream trend of traditional Chinese medicine compositions to play a therapeutic role. However, there is still a wide demand for the development of traditional Chinese medicine compositions with more definite efficacy and more reliable quality. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a traditional Chinese medicine composition with more definite efficacy and more reliable quality, wherein the traditional Chinese medicine composition comprises kudzu root extract and pine leaf extract.

[0006] Masson pine (Pinus massoniana Lamb.) belongs to the genus Pinus of the Pinaceae family and is one of the most widespread pine species in my country. It grows from Henan and southern Shandong in the north to Guangdong and Guangxi and Taiwan in the south, from the coast in the east to central Sichuan and Guizhou in the west, and is distributed throughout central and southern China. Masson pine leaves (or pine needles) have the functions of dispelling wind and promoting qi, promoting blood circulation and relieving pain, relaxing muscles and stopping bleeding. Pharmacological experiments have shown that masson pine leaves have antioxidant, anti-aging, antibacterial, cardiovascular protection, lipid regulation, and anti-tumor activities (Xiao Yunchuan et al., Study on the chemical composition of fresh masson pine leaves, Chinese Traditional and Herbal Drugs, 2015, 46(23): 3460-3463).

[0007] Pueraria root is the dried root of Pueraria lobata (Willd.) Ohwi, a leguminous plant. Its main chemical components are isoflavone compounds such as 3'-hydroxypuerarin, puerarin, 3'-methoxypuerarin, puerarin apiglycoside, daidzein, and daidzein. Modern pharmacological studies have shown that pueraria isoflavones have the effects of lowering blood sugar, lowering blood lipids, resisting thrombosis, enhancing immunity, anti-aging, and improving cardiovascular diseases (Suo Yaran et al., Study on the fingerprint and pattern recognition of pueraria root by high performance liquid chromatography, Global Chinese Medicine, 2019, 12(6): 844-849; Ye Bin et al., Study on the HPLC fingerprint of pueraria root medicinal materials and the determination of the contents of 6 components, Chinese Herbal Medicine, 2023, 54(24): 8222-8227).

[0008] Specifically, one aspect of the present invention is to provide a pharmaceutical composition comprising kudzu root extract and pine leaf extract, wherein puerarin, 3'-methoxypuerarin and puerarin apiglycoside account for 2-6wt% of the content of the pharmaceutical composition, and the mass ratio of puerarin, 3'-methoxypuerarin and puerarin apiglycoside is 15-45:2-9:4-12; catechins and massonoside B account for 0.2-0.7wt% of the content of the pharmaceutical composition, and the mass ratio of catechins to massonoside B is 0.2-2:1.5-7.

[0009] In some preferred embodiments, puerarin, 3'-methoxypuerarin and puerarin apioside in the pharmaceutical composition account for 3-6wt% of the content of the pharmaceutical composition, and the mass ratio of puerarin, 3'-methoxypuerarin and puerarin apioside is 20-45:3-9:5-11; catechins and massonoside B account for 0.2-0.6wt% of the content of the pharmaceutical composition, and the mass ratio of catechins and massonoside B is 0.2-2:1.5-6.

[0010] In some preferred embodiments, puerarin, 3'-methoxypuerarin and puerarin apioside in the pharmaceutical composition account for 3.5-6wt% of the content of the pharmaceutical composition, and the mass ratio of puerarin, 3'-methoxypuerarin and puerarin apioside is 22-45:3-9:5-11; catechins and massonoside B account for 0.2-0.6wt% of the content of the pharmaceutical composition, and the mass ratio of catechins and massonoside B is 0.5-2:1.5-6.

[0011] In some preferred embodiments, puerarin, 3'-methoxypuerarin and puerarin apioside in the pharmaceutical composition account for 3.5-5.5wt% of the content of the pharmaceutical composition, and the mass ratio of puerarin, 3'-methoxypuerarin and puerarin apioside is 22-40:4-9:6-11; catechin and massonoside B account for 0.3-0.6wt% of the content of the pharmaceutical composition, and the mass ratio of catechin and massonoside B is 0.5-2:2.5-5.5.

[0012] In some specific embodiments, the preparation method of the kudzu root and pine needle extract comprises: decocting fresh pine needle and kudzu root with water twice, filtering the decoction, combining the filtrate, concentrating, and spray drying.

[0013] In some specific embodiments, the preparation method of the kudzu root and pine needle extract comprises: respectively decocting fresh pine needle and kudzu root with water twice, filtering the decoction, combining the filtrate, concentrating, and spray drying.

[0014] In some preferred embodiments of the present invention, the quality of medicinal materials can be controlled by screening the origin of medicinal materials. There is a close connection between the origin of Chinese medicinal materials and the efficacy of drugs. Growth factors such as soil, altitude, climate (such as temperature, humidity, light), and growth environment will affect the chemical composition and efficacy of Chinese medicinal materials. For example, literature research shows that the content of 6 components in 32 batches of Pueraria medicinal materials from different origins was determined. The comprehensive score of Pueraria quality from different origins showed that Henan was the highest, followed by Shaanxi (Ye Bin et al., Study on HPLC fingerprint of Pueraria medicinal materials and determination of 6 components, Chinese Herbal Medicine, 2023, 54 (24): 8222-8227). Masson pine has a very wide distribution in my country. Zhang Xiaobo et al. sampled and analyzed Masson pine from more than 400 plots in 12 provinces across the country and found that the quality of pine needles in eastern Sichuan, Guizhou, Chongqing and other places was better (Zhang Xiaobo et al., Study on the suitability zoning of Masson pine production, Chinese Journal of Traditional Chinese Medicine, 2016, 41 (17): 3115-3121). Therefore, the technical solution of the present invention can be obtained by selecting medicinal materials from different origins with high content of puerarin or pine needles. In some preferred embodiments of the present invention, the inventors collect and identify medicinal materials from different origins, and preferably extract the effective ingredients from pueraria root produced in Shaanxi or Henan, and pine needles or pine needles of Masson pine in Sichuan and Chongqing.

[0015] In other preferred embodiments, the extraction efficiency and quality of the active ingredients in the medicinal materials can be improved by optimizing the process. For example, Gao Zhiping et al. found that the molecular structure of catechins, the active ingredient in pine needles or pine needles of Masson pine, has multiple phenolic hydroxyl groups that can form hydrogen bonds with the hydroxyl groups of organic solvents (such as ethanol) and have good solubility. Therefore, the ethanol extraction and reflux method is beneficial to the extraction of pine needle catechins (Gao Zhiping et al., Research on the Extraction Process of Catechins from Masson Pine Needles, Applied Chemical Industry, 2011, 40(3): 420-424). In some specific embodiments, the preparation method of the kudzu root and pine needle extract in the pharmaceutical composition of the present invention comprises: extracting fresh pine needles twice with 20-50% ethanol, decocting kudzu root with water twice, filtering the decoction, combining the filtrate, concentrating, and spray drying. In other specific embodiments, the content of the active ingredient in the extract can also be increased by adjusting the volume of the extraction solvent and / or the extraction time.

[0016] In other preferred embodiments, the content of active ingredients in the pharmaceutical composition can be increased by adjusting the dosage of fresh pine leaves or kudzu root medicinal materials.

[0017] The pharmaceutical composition of the present invention can be prepared by the preparation methods including but not limited to those listed above. As long as the components in the composition meet the above content ratio, the technical effects of the present invention can be achieved.

[0018] In other embodiments, the pharmaceutical composition further contains pearl powder.

[0019] In some specific implementation schemes, the dosage of fresh pine leaves, kudzu root and pearl powder is: 3600g fresh pine leaves, 600g kudzu root and 90g pearl powder.

[0020] In other specific embodiments, the dosage of fresh pine leaves, kudzu root and pearl powder is: 6000g fresh pine leaves, 1000g kudzu root and 90g pearl powder.

[0021] In some specific embodiments, the content determination method of puerarin, 3'-methoxypuerarin and puerarin apiglycoside is: high performance liquid chromatography, octadecylsilane bonded silica gel as filler, acetonitrile as mobile phase A, 0.2% acetic acid solution as mobile phase B, detection wavelength of 250nm, gradient elution according to the following table;

[0022]

[0023] In some specific embodiments, the content of catechins and massonoside B is determined by high performance liquid chromatography, using octadecylsilane bonded silica gel as a filler, methanol as a mobile phase A, 0.1% phosphoric acid aqueous solution as a mobile phase B, a detection wavelength of 280 nm, and gradient elution according to the following table:

[0024]

[0025] In other embodiments, the pharmaceutical composition further contains other pharmaceutically acceptable excipients.

[0026] In some embodiments, the pharmaceutically acceptable excipients include diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), and the like.

[0027] In some embodiments, the pharmaceutical composition can be prepared into a dosage form such as capsule, pill, granule, oral liquid, injection, powder, tablet, lozenge or lozenge, and a suitable pharmaceutical carrier in the art can be selected for different dosage forms. The preferred dosage form is capsule, tablet, pill, granule, oral liquid, and the most preferred dosage form is capsule.

[0028] Another aspect of the present invention is to provide a method for preparing the pharmaceutical composition of the present invention, comprising the steps of adding pearl layer powder and other pharmaceutically acceptable excipients to kudzu root and pine leaf extracts to obtain the pharmaceutical composition.

[0029] Another aspect of the present invention is to provide use of the pharmaceutical composition of the present invention in the preparation of a medicament for preventing or treating hypertension and / or hyperlipidemia.

[0030] In some preferred embodiments, the pharmaceutical composition of the present invention is used to prepare a drug for preventing or treating hypertension and hyperlipidemia.

[0031] The pharmaceutical composition of the present invention has the following beneficial effects: the pharmaceutical composition of the present invention can effectively play the role of lowering blood pressure and blood lipids, and its therapeutic effect is more accurate and reliable than that of the prior art. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The dose-effect curve of the vasodilation response under the action of the pharmaceutical composition in Example 1;

[0033] Figure 2 The dose-effect curve of the vasodilation response under the action of the pharmaceutical composition in Example 2;

[0034] Figure 3 The triglyceride content of the pharmaceutical composition in Example 3 in the zebrafish experiment;

[0035] Figure 4Cholesterol content of the pharmaceutical composition in Example 3 in zebrafish experiment. DETAILED DESCRIPTION

[0036] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of protection claimed in this application, and those skilled in the art can make various changes and modifications to the invention of this application based on the disclosed content, and they should also fall within the scope of protection claimed in this application.

[0037] Unless otherwise specified, the various chemical reagents used in the embodiments of the present invention are obtained through conventional commercial channels or prepared by themselves according to existing technologies, and do not limit the present invention.

[0038] Medicinal materials and reagents

[0039] Pinus massoniana: origin of medicinal materials, Lishui, Zhejiang

[0040] Pinus massoniana: origin of medicinal materials, eastern Sichuan

[0041] Pueraria root: origin of medicinal materials, Guangdong

[0042] Pueraria root: origin of medicinal materials, Shaanxi Methanol: Chengdu Chuangrui Chemicals Co., Ltd. Ethanol: Chengdu Chuangrui Chemicals Co., Ltd. Acetic acid: Chengdu Chuangrui Chemicals Co., Ltd. Acetonitrile: Chengdu Chuangrui Chemicals Co., Ltd.

[0043] Puerarin: China Food and Drug Administration, 96.8%

[0044] 3'-Methoxypuerarin: Lemet Pharmaceuticals, 99.89%

[0045] Puerarin apigenin glycoside: Lemet Pharmaceuticals, 99.26%

[0046] Catechins: China Food and Drug Administration, 95.1%

[0047] Preparation of Massonoside B:

[0048] Take an appropriate amount of fresh pine leaves of Masson pine, extract with 50% ethanol under heating and reflux, combine the extracts, and concentrate under reduced pressure to obtain an extract. After the extract is dispersed in water, it is extracted with n-butanol to obtain an n-butanol extract extract. Take the n-butanol extract, adsorb water elution with macroporous resin D101, and then gradient elute through SBC MCI column (20% ethanol, 50% ethanol), collect the 50% ethanol elution part and concentrate to obtain Pr.3A3. Pr.3A3 is passed through Sephadex G25 column (water elution) to obtain 3 parts Pr.3A3A~Pr.3A3C. Pr.3A3B is gradient eluted through SBC MCI column (30%, 40% ethanol), collect the 40% ethanol elution part and concentrate to obtain Pr.Pr.3A3B3. Pr.Pr.3A3B3 was purified by two silica gel column chromatography with dichloromethane-ethanol (5:1) and petroleum ether-acetone (1:3) to obtain a white powder. 1 H-NMR, 13 Identification of massonoside B by C-NMR.

[0049] 1 H-NMR (400MHz, DMSO-d6) δ: 9.10 (1H, s, ph-3′-OH), 7.06 (1H, d, J = 8.4Hz, H-5), 7.03 (1H, d, J = 1.2Hz, H-2), 6.90 (1H, dd, J = 8.4, 1.2Hz, H-5), 6.53 (1H, s,H-6′),6.50(1H,s,H-2′),5.46(1H,d,J=6.0Hz,H-7),5.25(1H,s,H-1″) ,5.03(1H,t,J=5.2Hz,9-OH),5.00(1H,d,J=4.4Hz,2″-OH),4.86(1H,d,J=5 .6Hz,4″-OH),4.73(1H,d,J=6.0Hz,3″-OH),4.43(1H,t,J=5.2Hz,9′-OH), 3.77(3H,s,3-OMe),3.72(1H,dd,J=10.4,5.2Hz,H-9a),3.64(1H,m,H-9b), 3.59(2H,m,H-2″,8),3.42(4H,m,H-3″,5″,9′),3.27(1H,dt,J=9.2,5.6Hz ,H-4″),2.45(2H,m,H-7′),1.65(2H,m,H-8′),1.10(1H,d,J=6.0Hz,H-6″); 13C-NMR(100MHz,DMSO-d6)δ:150.0(C-4),144.7(C-3),144.6(C-4′),140.7(C-3′),136.7( C-5′),134.9(C-1),128.7(C-1′),118.0(C-6),117.8(C-6′),115.8(C-2′),114.9(C-5),1 10.5(C-2),99.6(C-1″),86.1(C-7),71.7(C-4″),70.4(C-3″),70.2(C-2″),69.6(C-5″), 63.2(C-9), 60.2(C-9′), 56.0(3-OMe), 53.7(C-8), 34.7(C-7′), 31.3(C-8′), 17.8(C-6″).

[0050] Determination method of puerarin, 3'-methoxypuerarin and puerarin apiglycoside:

[0051] Measured according to high performance liquid chromatography (General Rule 0512). Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler (100 mm × 2.1 mm, 1.9 μm, Thermo Hypersil GOLD C 18 ); acetonitrile as mobile phase A, 0.2% acetic acid solution as mobile phase B. Gradient elution was performed according to the following table; flow rate 0.3 ml per minute; column temperature 30°C; detection wavelength 250 nm; theoretical plate number calculated based on puerarin should be no less than 4000.

[0052] Table 1 Gradient elution program

[0053]

[0054] Preparation of reference solution: Take appropriate amount of puerarin, 3'-methoxypuerarin, and puerarin apiglycoside reference substances, accurately weigh them, and add methanol to prepare 0.2mg / ml reference substance mother solution. Accurately measure appropriate amount of each mother solution, add methanol to prepare mixed reference substance solutions containing 50μg, 50μg, and 60μg of puerarin, 3'-methoxypuerarin, and puerarin apiglycoside per 1ml.

[0055] Preparation of test solution: About 0.1 g of the product content is accurately weighed and placed in a conical flask. 50 ml of methanol is accurately added and the total amount is weighed. Ultrasonic treatment (power 250 W, frequency 45 kHz) is performed for 30 minutes. The solution is cooled and weighed again. The weight is supplemented with methanol to make up for the loss. The solution is shaken well and filtered. The filtrate is taken to obtain the solution.

[0056] Determination method: Accurately pipette 1 μl of reference solution and test solution respectively, inject into liquid chromatograph, and determine.

[0057] Measured according to high performance liquid chromatography (General Rule 0512). Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler (150 mm × 2.1 mm, 1.7 μm, C 18 ); methanol was used as mobile phase A, 0.1% phosphoric acid aqueous solution was used as mobile phase B, the flow rate was 0.2 ml / min, the column temperature was 30°C, the detection wavelength was 280 nm, and gradient elution was performed according to the table below.

[0058]

[0059]

[0060] Preparation of control solution: Take appropriate amount of catechin and pine glycoside B reference substances, weigh them accurately, and add 40% ethanol to make 0.2mg / ml reference substance mother solution. Accurately measure appropriate amount of each mother solution, add methanol to make mixed reference substance solution containing 40ug and 80ug per 1ml, accurately measure 1, 2, 3, 4, 5ml, put them in 5ml volumetric flask with 40% ethanol to dilute to the scale and shake well. Preparation of test solution: Take the contents of this product, grind them finely, mix them well, take 0.2g, weigh them accurately, put them in a stoppered conical flask, accurately add 25ml of 40% ethanol, and weigh them. Ultrasonic treatment (power 250W, frequency 45KHz) for 20 minutes, cool, make up for the weight loss, and you are done.

[0061] Determination method: Accurately pipette 1ul of reference solution and test solution respectively, inject into liquid chromatograph, determine, and calculate using the standard curve method.

[0062] Example 1

[0063] 1.1 Preparation method

[0064] Experimental group 1: The drug preparation was prepared according to the following prescription and process, wherein the prescription was:

[0065] 3600g fresh pine leaves (Lishui, Zhejiang)

[0066] Pueraria root 600g (Guangdong)

[0067] Pearl layer powder 90g

[0068] Preparation method: Take the above three ingredients, pine needles and kudzu root medicinal materials in the above dosage ratio, add water and decoct twice, add 8 times the amount of water for the first time, decoct for 1.5 hours, add 6 times the amount of water for the second time, decoct for 1.5 hours, combine the decoctions, filter, and decompress the filtrate to form an extract with a relative density of 1.2 at 70°C, spray dry, add pearl layer powder and appropriate amounts of starch, talcum powder and magnesium stearate, mix well, and obtain powder. Then take an appropriate amount of powder and determine the content of puerarin, 3'-methoxypuerarin, puerarin apiglycoside, catechin and scutellarin B in the powder according to the method of the present invention. The results are shown in Table 3:

[0069] Table 3

[0070]

[0071] Experimental group 2: Based on the powder prepared in combination 1, an appropriate amount of puerarin, 3'-methoxypuerarin, puerarin apiglycoside, catechin or massonoside B monomer compound was added to prepare the pharmaceutical composition described in Table 4:

[0072] Table 4

[0073]

[0074] 1.2 Experiment on tension effect of rat mesenteric artery Experimental purpose: To verify the effect of drug composition of different combinations on the contractile and diastolic function of rat mesenteric artery.

[0075] Experimental animals: Male SD rats, SPF grade, body weight 220-440g.

[0076] Sample solution preparation: 5.0 g of the powder of the different pharmaceutical compositions in Example 1 was dissolved in 22 mL of benchtop solution to obtain a solution with a concentration of 57 mg / ml.

[0077] Preparation of intestinal absorption solution: Rats were fasted for 12 hours (fasted overnight) before the experiment and were killed by dislocating the neck. Measure 14 cm downward from 10 cm below the pylorus of the stomach as the jejunal segment, measure 14 cm at intervals of 10 cm as the second experimental intestinal segment, measure 14 cm at intervals of 10 cm as the third experimental intestinal segment, and measure 14 cm upward from 5 cm above the ileocecal valve as the fourth experimental intestinal segment. Put the cut intestinal tube into 0℃ Tyrode's solution for flushing until there is no intestinal content, and carefully turn the intestinal segment. Rinse the surface with 0℃ Tyrode's solution, tie one end with silk thread to form a sac, and put the other end on a homemade silicone hose and tie it to fix. Inject 2ml of Tyrode's solution into the intestinal tube and carefully place it in the bath tube. 22mL of Tyrode's solution was injected into the bath tube of the blank group; 22mL of different combinations of drug solutions were injected into the bath tube of the drug group. 95% CO2 and 5% O2 were continuously introduced into the bath tubes of the blank group and the drug group, and the tubes were taken out after being bathed in 37℃ constant temperature water for 2 hours. Carefully cut open the upper end of the intestinal tube, pour out the liquid in the intestinal sac, filter with a 0.22μm nylon filter membrane, and store at -20℃ for later use.

[0078] Preparation of isolated vascular rings

[0079] Rats were anesthetized, and the mesenteric plexus was removed and placed in cold Krebs solution. The secondary mesenteric artery was isolated under a microscope. The processed artery was cut into a vascular ring of about 2 mm. The processed vascular ring was placed in a bath containing 5 ml of Krebs solution, maintained at a constant temperature of 37°C and continuously aerated with 95% CO2 and 5% O2, and the tension of the vascular ring was finely adjusted to the baseline value. After the secondary mesenteric artery was stable, KCl (60 mol / L) was added to stimulate it to stimulate the maximum contraction and stabilize the vascular contraction state.

[0080] Analytical methods

[0081] After the blood vessels were pre-contracted with KCl and stabilized, the cumulative drug addition method was used to add blank intestinal absorption solution and drug-containing intestinal absorption solution of compositions 1-4 to the bath to 200μl, 400μl, and 600μl, respectively, and added once every 3 minutes. The final dose observation was extended to 10min. The effects of drug intestinal absorption solutions of different compositions on the tension of rat mesenteric secondary arteries were observed. The maximum contraction amplitude induced by KCl (60mol / L) was 100%, and the ratio of the vascular tension relaxation amplitude after adding the drug composition to the maximum contraction amplitude induced by KCl was the dose-effect curve (relaxation rate) of the vascular relaxation response, expressed as a percentage. The average relaxation rate and standard deviation SD value (n=4) were calculated, and the results are shown in Table 5 and Figure 1 shown.

[0082] Table 5

[0083]

[0084] At the same time, the percentage increase in the relaxation rate of the drug group relative to the blank intestinal fluid group under the condition of adding a volume of 600 μl was calculated according to the following formula. The results are shown in Table 6.

[0085] Diastolic rate increase percentage = (diastolic rate of blank group - diastolic rate of drug group) / diastolic rate of blank group * 100%

[0086] Table 6

[0087] Composition 600μl relaxation rate increase percentage Blank control / Composition 1 38.06％ Composition 2 266.92％ Composition 3 269.89％ Composition 4 925.91％

[0088] Conclusion: The above experiments show that drug composition groups 2-4 can significantly enhance the vasodilation effect compared with the blank control group and drug composition 1.

[0089] Example 2

[0090] 2.1 Preparation method

[0091] Based on the powder of composition 1 in Example 1, an appropriate amount of puerarin, 3'-methoxypuerarin, puerarin apiglycoside, catechin or massonoside B monomer compound is added to prepare the pharmaceutical composition described in Table 7:

[0092] Table 7

[0093]

[0094]

[0095] 2.2 Experiment on tension effect of rat mesenteric artery

[0096] The experimental method is the same as in Example 1, and the results are shown in Table 8 and Figure 2 shown.

[0097] Table 8

[0098]

[0099] The results of the percentage increase in the relaxation rate of the drug group relative to the blank intestinal fluid group under the condition of a volume of 600 μl are shown in Table 9.

[0100] Table 9

[0101]

[0102]

[0103] Conclusion: The above experiments show that the drug composition groups 5-13 can significantly enhance the vasodilation effect compared with the blank control group.

[0104] Example 3: Lipid-lowering efficacy test in zebrafish hyperlipidemia model

[0105] Zebrafish strain: Albino strain with melanin allele mutation; wild-type AB strain. Experimental groups: normal control group, model control group, positive control group (atorvastatin calcium), drug composition sample group; wherein the preparation method of the drug composition group includes taking 1.0g of the powder of each composition, preparing it into a 2.0mg / ml mother solution with ultrapure water, and storing it at 4°C. Atorvastatin calcium is prepared into a 11.6mg / ml mother solution with DMSO, and stored in subpackages at -20°C. When administering, the drug composition and atorvastatin calcium are diluted to the required concentration using standard dilution water.

[0106] Experimental method: Zebrafish fry were incubated until 5 days post fertilization (dpf), healthy embryos were selected and randomly divided into groups, with 30 fry in each group. The positive control group was given atorvastatin calcium (11.6 μg / mL), and the drug composition sample group was given the drug composition described in Example 1 or 2 (1000 μg / mL). The normal control group was given fish water, and the other groups were fed with high-sugar and high-fat feed. After 48 hours of treatment, zebrafish samples were collected, with 3 parallels in each group, and the triglyceride content in zebrafish was detected using a triglyceride kit; the cholesterol content in zebrafish was detected using a cholesterol kit.

[0107] Experimental results: The triglyceride and cholesterol contents of each group were as follows Figure 3 and Figure 4 Wherein *p<0.05, **p<0.01, ***p<0.001 represent the difference level with the model control group; #p<0.05, ##p<0.01, ###p<0.001 represent the difference level with the composition 1 and other sample groups.

[0108] Example 4

[0109] The pharmaceutical preparation is prepared according to the following prescription and process, wherein the prescription is:

[0110] 3600g fresh pine leaves (Eastern Sichuan)

[0111] Pueraria root 600g (Shaanxi)

[0112] Pearl layer powder 90g

[0113] Preparation method: Take the above three ingredients, take the pine needles and kudzu root medicinal materials in the above dosage ratio, add water and decoct twice, add 8 times the amount of water for the first time, decoct for 1.5 hours, add 6 times the amount of water for the second time, decoct for 1.5 hours, combine the decoctions, filter, and decompress the filtrate to form an extract with a relative density of 1.2 at 70°C, spray dry, add pearl layer powder and appropriate amounts of starch, talcum powder and magnesium stearate, mix well, and obtain powder. Then take an appropriate amount of powder and determine the content of puerarin, 3'-methoxypuerarin, puerarin apiglycoside, catechin and scutellarin B in the powder according to the method described in the present invention. The results are shown in Table 10:

[0114] Table 10

[0115]

[0116]

[0117] Example 5

[0118] The pharmaceutical preparation is prepared according to the following prescription and process, wherein the prescription is:

[0119] 3600g fresh pine leaves (Lishui, Zhejiang)

[0120] Pueraria root 600g (Guangdong)

[0121] Pearl layer powder 90g

[0122] Preparation method: Take the fresh pine leaves with the above dosage ratio, decoct twice with 20% ethanol solution, add 8 times the amount of ethanol solution for 2 hours for the first time, and add 6 times the amount of ethanol solution for 2 hours for the second time; at the same time, take the Pueraria root with the above dosage ratio, add water and decoct twice, add 10 times the amount of water for the first time, decoct for 3 hours, and add 10 times the amount of water for the second time, decoct for 3 hours; combine the decoctions, filter, and decompress the filtrate to form an extract with a relative density of 1.2 at 70°C, spray dry, add pearl layer powder and appropriate amounts of starch, talcum powder and magnesium stearate, mix well, and obtain powder. Then take an appropriate amount of powder and determine the content of puerarin, 3'-methoxypuerarin, puerarin apiglycoside, catechin and scutellarin B in the powder according to the method of the present invention. The results are shown in Table 11:

[0123] Table 11

[0124]

[0125] Example 6

[0126] The pharmaceutical preparation is prepared according to the following prescription and process, wherein the prescription is:

[0127] 3600g fresh pine leaves (Lishui, Zhejiang)

[0128] Pueraria root 600g (Guangdong)

[0129] Pearl layer powder 90g

[0130] Preparation method: Take the fresh pine leaves with the above dosage ratio, decoct twice with 50% ethanol solution, add 8 times the amount of ethanol solution for the first time and decoct for 2h, add 6 times the amount of ethanol solution for the second time and decoct for 2h; at the same time, take the Pueraria root with the above dosage ratio, add water and decoct twice, add 8 times the amount of water for the first time and decoct for 2 hours, add 8 times the amount of water for the second time and decoct for 2 hours; combine the decoctions, filter, and decompress the filtrate to form an extract with a relative density of 1.2 at 70°C, spray dry, add pearl layer powder and appropriate amounts of starch, talcum powder and magnesium stearate, mix well, and obtain powder. Then take an appropriate amount of powder and determine the content of puerarin, 3'-methoxypuerarin, puerarin apiglycoside, catechin and scutellarin B in the powder according to the method of the present invention. The results are shown in Table 12:

[0131] Table 12

[0132]

[0133] Example 7

[0134] The pharmaceutical preparation is prepared according to the following prescription and process, wherein the prescription is:

[0135] Fresh pine leaves 6000g (Lishui, Zhejiang)

[0136] Pueraria root 1000g (Guangdong)

[0137] Pearl layer powder 90g

[0138] Preparation method: Take the above three ingredients, pine needles and kudzu root medicinal materials in the above dosage ratio, add water and decoct twice, add 10 times the amount of water for the first time, decoct for 2 hours, add 8 times the amount of water for the second time, decoct for 2 hours, combine the decoctions, filter, and decompress the filtrate to form an extract with a relative density of 1.2 at 70°C, spray dry, add pearl layer powder and appropriate amounts of starch, talcum powder and magnesium stearate, mix well, and obtain powder. Then take an appropriate amount of powder and determine the content of puerarin, 3'-methoxypuerarin, puerarin apiglycoside, catechin and scutellarin B in the powder according to the method of the present invention. The results are shown in Table 13:

[0139] Table 13

[0140]

[0141] Example 8

[0142] The effect of the drug composition of Compositions 14-17 on the systolic and diastolic function of rat mesenteric arteries was verified by a rat mesenteric artery tension experiment. The experimental method was the same as that of Example 1. The results are shown in Table 14.

[0143] Table 14

[0144]

[0145] The results of the percentage increase in the relaxation rate of the drug group relative to the blank intestinal fluid group under the condition of a volume of 600 μl are shown in Table 15.

[0146] Table 15

[0147] Composition 600μl relaxation rate increase percentage Blank control / Composition 14 1072.69％ Composition 15 1049.32％ Composition 16 989.23％ Composition 17 1040.85％

[0148] Conclusion: The above experiments show that drug combination groups 14-17 can significantly enhance the vasodilation effect compared with the blank control group.

[0149] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A pharmaceutical composition comprising kudzu root extract and pine leaf extract, characterized in that: In the pharmaceutical composition, puerarin, 3'-methoxypuerarin and puerarin apiglycoside account for 2-6wt% of the pharmaceutical composition, and the mass ratio of puerarin, 3'-methoxypuerarin and puerarin apiglycoside is 15-45:2-9:4-12; catechin and massonoside B account for 0.2-0.7wt% of the pharmaceutical composition, and the mass ratio of catechin and massonoside B is 0.2-2:1.5-7.

2. The pharmaceutical composition according to claim 1, characterized in that The puerarin, 3'-methoxypuerarin and puerarin apiglycoside account for 3-6wt% of the content of the pharmaceutical composition, and the mass ratio of puerarin, 3'-methoxypuerarin and puerarin apiglycoside is 20-45:3-9:5-11; the catechin and massonoside B account for 0.2-0.6wt% of the content of the pharmaceutical composition, and the mass ratio of catechin and massonoside B is 0.2-2:1.5-6.

3. The pharmaceutical composition according to claim 1, characterized in that The puerarin, 3'-methoxypuerarin and puerarin apiglycoside account for 3.5-6wt% of the content of the pharmaceutical composition, and the mass ratio of puerarin, 3'-methoxypuerarin and puerarin apiglycoside is 22-45:3-9:5-11; the catechin and massonoside B account for 0.2-0.6wt% of the content of the pharmaceutical composition, and the mass ratio of catechin and massonoside B is 0.5-2:1.5-6.

4. The pharmaceutical composition according to claim 1, characterized in that The puerarin, 3'-methoxypuerarin and puerarin apiglycoside account for 3.5-5.5wt% of the content of the pharmaceutical composition, and the mass ratio of puerarin, 3'-methoxypuerarin and puerarin apiglycoside is 22-40:4-9:6-11; the catechin and massonoside B account for 0.3-0.6wt% of the content of the pharmaceutical composition, and the mass ratio of catechin and massonoside B is 0.5-2:2.5-5.

5.

5. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition also contains pearl layer powder.

6. The pharmaceutical composition according to claim 1, characterized in that The pharmaceutical composition also contains other pharmaceutically acceptable excipients.

7. The pharmaceutical composition according to claim 6, characterized in that The pharmaceutical composition is a capsule, a pill, a granule or a tablet; preferably, the pharmaceutical composition is a capsule.

8. The pharmaceutical composition according to any one of claims 1 to 7, characterized in that The preparation method of the kudzu root and pine needle extract comprises: decocting fresh pine needles and kudzu root twice, filtering the decoction, combining the filtrates, concentrating, and spray drying to obtain the extract; or, decocting fresh pine needles and kudzu root twice, filtering the decoction, combining the filtrates, concentrating, and spray drying to obtain the extract.

9. The pharmaceutical composition according to any one of claims 1 to 7, characterized in that The preparation method of the kudzu root and pine needle extract comprises: extracting fresh pine needles twice by decocting with 20-50% ethanol, decocting kudzu root twice by adding water, filtering the decoction, combining the filtrate, concentrating and spray drying.

10. The pharmaceutical composition according to claim 8 or 9, characterized in that The preparation method of the pharmaceutical composition comprises: adding pearl layer powder and other pharmaceutically acceptable auxiliary materials into kudzu root extract and pine leaf extract to obtain the pharmaceutical composition.

11. The pharmaceutical composition according to claim 10, characterized in that The dosage of fresh pine leaves, kudzu root and pearl powder is: 3600g fresh pine leaves, 600g kudzu root and 90g pearl powder.

12. The pharmaceutical composition according to claim 10, characterized in that The dosage of fresh pine leaves, kudzu root and pearl powder is: 6000g fresh pine leaves, 1000g kudzu root and 90g pearl powder.

13. The pharmaceutical composition according to claim 1, characterized in that The content determination method of puerarin, 3'-methoxypuerarin and puerarin apiglycoside is as follows: high performance liquid chromatography, using octadecylsilane bonded silica gel as filler, acetonitrile as mobile phase A, 0.2% acetic acid solution as mobile phase B, detection wavelength of 250nm, and gradient elution according to the following table; 14. The pharmaceutical composition according to claim 1, characterized in that The content determination method of catechins and spicroside B is: high performance liquid chromatography, octadecylsilane bonded silica gel as filler, methanol as mobile phase A, 0.1% phosphoric acid aqueous solution as mobile phase B, detection wavelength of 280nm, gradient elution according to the following table:

15. Use of the pharmaceutical composition according to any one of claims 1 to 14 in the preparation of a medicament for preventing or treating hypertension and / or hyperlipidemia; preferably, use of the pharmaceutical composition according to any one of claims 1 to 14 in the preparation of a medicament for preventing or treating hypertension and / or hyperlipidemia.

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