Preparation method of pharmaceutical composition

Abstract

The invention discloses a preparation method of a pharmaceutical composition, and relates to the field of biological medicines. The pharmaceutical composition is prepared from the following raw materials in parts by weight: 40-60 parts of spica prunellae, 20-30 parts of flos chrysanthemi, 20-30 parts of semen cassiae and 20-30 parts of tartary buckwheat. The preparation method of the composition comprises the following steps: taking supernate from the selfheal and the chrysanthemum by adopting a traditional water decoction process, taking supernate from the cassia seed and the tartary buckwheat by adopting an alcohol extraction process, and concentrating, drying and uniformly mixing the two parts of supernate to obtain the blood fat reducing composition. A mouse test hypertension model and a caenorhabditis elegans antioxidant test prove that the composition can remarkably prolong the life of the caenorhabditis elegans and has a remarkable antioxidant effect. The composition can be used for preparing hypolipidemic drugs or health-care products, wherein the drugs or health-care products can be prepared into capsules from the composition and pharmaceutics.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a method for preparing a pharmaceutical composition. Background Art

[0002] Hypertension is a complex clinical syndrome with abnormal increase of systemic blood pressure exceeding 140 / 90 mmHg as the main clinical feature. It has the characteristics of a wide range of patients, unclear pathogenesis and hidden disease. The "China Cardiovascular Health and Disease Report 2019" pointed out that the number of hypertensive patients in my country exceeds 245 million, accounting for about 15% to 25% of the total population.

[0003] Arterial hypertension is one of the main risk factors for atherosclerosis, which ultimately leads to thromboembolic events such as myocardial infarction or stroke, and is therefore the leading cause of global mortality and severe disability. Intracranial arteriolar remodeling is the main adaptive mechanism of chronic high blood pressure. However, atherosclerosis can lead to changes in the function of vascular smooth muscle, which is a key cell in the process of vascular remodeling. VSMCs in mature blood vessels are highly plastic cells that can respond to local environmental changes and change their phenotype. In addition, studies have shown that excessive inflammatory response is an important pathological feature of hypertension, and inhibiting inflammatory response can improve the course of hypertension. In summary, hypertension has a high incidence rate in the population and a complex pathogenesis, but currently the main clinical treatment for it is the following categories: diuretics, beta blockers, calcium antagonists, etc. Long-term use of these drugs will affect the patient's electrolyte balance, lower limb edema, headache, dizziness and other side effects, and it is difficult for patients to adhere to medication for a long time.

[0004] Prunella vulgaris, chrysanthemum, cassia seed, and bitter buckwheat all have the effects of anti-atherosclerosis, anti-inflammation, enhancing immunity, protecting the human cardiovascular system, and removing oxygen free radicals in the body. The mechanism of action of lowering blood pressure involves a wide range. Prunella vulgaris is a commonly used tonic medicinal material in traditional Chinese medicine formulas, which has the effect of delaying aging. Chrysanthemum and cassia seed both have the effects of inhibiting the secretion of chronic inflammatory factors and inhibiting the level of oxidative stress. Bitter buckwheat can improve a variety of cardiovascular and pulmonary diseases, thereby delaying the occurrence and development of hypertension.

[0005] Most of the existing Chinese herbal medicines are taken as a single dose in water or boiled in water, and the effect is difficult to fully realize. It is necessary to fully tap the relevant herbal resources, screen the combination formula experiments, exert the synergistic efficacy, and avoid the drug antagonism at the same time, and develop a combination with significant blood pressure lowering effect, simple composition, convenient long-term use, high safety, fewer adverse reactions, and easy to be accepted by patients. Summary of the invention

[0006] The object of the present invention is to provide a method for preparing a pharmaceutical composition. The prepared pharmaceutical composition has significant blood pressure lowering effect, simple composition, convenience for long-term use, high safety, few adverse reactions, and is easily accepted by patients.

[0007] A method for preparing a pharmaceutical composition, characterized in that the pharmaceutical composition is made of the following raw materials in parts by weight: 40-60 parts of selfheal, 20-30 parts of chrysanthemum, 20-30 parts of cassia seeds, and 20-30 parts of tartary buckwheat.

[0008] The pharmaceutical composition of the present invention is prepared from the following raw materials in parts by weight: 60 parts of Prunella Vulgaris, 30 parts of chrysanthemum, 30 parts of cassia seed and 30 parts of tartary buckwheat.

[0009] The preparation method of a pharmaceutical composition described in the present invention comprises the following steps: weighing corresponding weight portions of herba selfhei, chrysanthemum, cassia seed and tartary buckwheat, taking supernatant liquor from herba selfhei and chrysanthemum by traditional water decoction process, taking supernatant liquor from cassia seed and tartary buckwheat by alcohol extraction process, concentrating, drying and mixing the two parts of supernatant liquor to obtain the pharmaceutical composition.

[0010] The preparation method of a pharmaceutical composition of the present invention comprises the following steps: a decoction process of Prunella vulgaris and chrysanthemum is as follows: 100g of the medicinal materials are added with 6-8L of water, soaked for 10-12 hours, and then extracted at 90-100°C for 1.5-2.5 hours;

[0011] 50g of Cassia seed and 50g of Tartary Buckwheat were added to 6-8L of 95% ethanol, soaked for 4-6 hours, and then extracted at 80°C for 1.5-2.5 hours;

[0012] The two parts of supernatant are concentrated, dried and mixed to obtain a pharmaceutical composition.

[0013] Application of the pharmaceutical composition prepared by the method of the present invention in lowering blood pressure.

[0014] The method of the present invention has positive application effects:

[0015] Prunella vulgaris, chrysanthemum, cassia seed, and bitter buckwheat all have the effects of anti-atherosclerosis, anti-inflammation, enhancing immunity, protecting the human cardiovascular system, and removing oxygen free radicals in the body. The mechanism of action of lowering blood pressure involves a wide range. Prunella vulgaris is a commonly used tonic medicinal material in traditional Chinese medicine formulas, which has the effect of delaying aging. Chrysanthemum and cassia seed both have the effects of inhibiting the secretion of chronic inflammatory factors and inhibiting the level of oxidative stress. Bitter buckwheat can improve a variety of cardiovascular and pulmonary diseases, thereby delaying the occurrence and development of hypertension.

[0016] Most of the existing Chinese herbal medicines are taken as a single dose in water or boiled in water, and the effect is difficult to fully realize. It is necessary to fully tap the relevant herbal resources, screen the combination formula test, exert the synergistic efficacy, and avoid the drug antagonism at the same time, and develop a drug composition that has a significant blood pressure lowering effect, simple composition, convenient long-term use, high safety, fewer adverse reactions, and is easily accepted by patients.

[0017] The drug composition screened by a large number of preparation method experiments and studies in the present invention has passed the mouse hypertension test and the nematode antioxidant test, proving that the composition can significantly reduce the blood pressure of mice and prolong the life span of nematodes, and has a significant antioxidant effect.

[0018] The pharmaceutical composition can reduce the systolic and diastolic blood pressure of hypertension model mice, and simultaneously reduce the SOD level of the Caenorhabditis elegans antioxidant test model Caenorhabditis elegans, and has the effects of inhibiting the secretion of chronic inflammatory factors and inhibiting the level of oxidative stress, thereby improving cardiovascular diseases and delaying the occurrence and development of hypertension. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 Aortic fibrosis in experimental mice (Masson staining, ×400)

[0020] A: normal control group; B: model group; C: drug combination group.

[0021] Figure 2 Flow cytometry to detect macrophage polarization

[0022] A: Effect of the drug composition on M1 polarization of macrophages. B: Effect of the drug composition on M2 polarization of macrophages. DETAILED DESCRIPTION

[0023] The present invention is further described in conjunction with specific embodiments, but these embodiments are only intended to illustrate the present invention and are not intended to limit the scope of the present invention.

[0024] Example 1

[0025] Preparation of a pharmaceutical composition:

[0026] Take 60g of selfheal, 30g of chrysanthemum, 30g of cassia seed and 30g of bitter buckwheat; add 6L of water per 100g of selfheal and chrysanthemum medicinal materials, soak for 12 hours, extract for 2 hours at 100℃, stand for 2 hours after the extraction, take the supernatant, concentrate the water extract to a relative density of 1.10-1.15, and transfer it to an iron plate for freeze drying; add 8L of 95% ethanol per 100g of cassia seed and bitter buckwheat medicinal materials, soak for 5 hours, extract for 2 hours at 75℃, stand for 2 hours after the extraction, take the supernatant, concentrate the water extract to a relative density of 1.10-1.15, and transfer it to an iron plate for freeze drying; crush the two parts of dried products, mix them evenly, and obtain a pharmaceutical composition.

[0027] Example 2

[0028] Establishment of hypertension model and detection of related indicators

[0029] 2.1 Experimental instruments and reagents

[0030] Angiotensin II (Ang II) was purchased from Sigma, USA. Capsule micro-osmotic pump was purchased from ALZET, USA. IL-1, IL-6, tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β) kits were purchased from ThermoFisher, USA. Anti-CD80 antibody, anti-CD86 antibody, anti-CD163 antibody and anti-CD206 antibody were purchased from RD, USA.

[0031] 2.2 Establishment of hypertension model

[0032] According to the random number table method, 30 C57 / B6 mice were divided into 3 groups: control group, model group and drug combination group, with 10 mice in each group. The hypertension model was established by continuous infusion of Ang II via capsule micro-osmotic pump implantation, where the mice in the Ang II group were infused with Ang II at a dose of 750 ng / (kg·min), and the mice in the control group were infused with an equal amount of normal saline.

[0033] 2.3 Antioxidant assay in Caenorhabditis elegans

[0034] By adding paraquat to the culture medium, a Caenorhabditis elegans oxidative damage model can be well established. In this study, the antioxidant effect of the pharmaceutical composition of the present invention and its protective effect against paraquat damage were verified by using a Caenorhabditis elegans antioxidant test.

[0035] 2.4 Blood pressure measurement

[0036] The blood pressure changes of mice were detected using a small animal non-invasive blood pressure meter one day before perfusion, and on the first, seventh, 14th, 21st, and 28th days after perfusion. The operation was as follows: the heating device was turned on 15 minutes in advance, the mice were placed in a long cylindrical bucket and fixed, and placed on the heating plate. The small animal non-invasive blood pressure meter was turned on, the sensor was placed on the base of the mouse tail, and the signal of the pressure sensor was collected. Each mouse was measured at least 5 times and the average value was taken.

[0037] 2.5 Serological testing

[0038] Peripheral blood was collected 28 days after Ang II perfusion, and serum was collected after centrifugation. The levels of IL-1, IL-6, TNF-α, and TGF-β in mouse serum were detected according to the kit instructions.

[0039] 2.6 Vascular activity measurement

[0040] After killing the mice, the chest cavity was opened, the aorta was isolated, and quickly transferred to pre-cooled Kleinstein solution. The aorta was fixed and placed in an organ bath, connected to the steel wire on the sensor, and different concentrations of phenylephrine and acetylcholine were added to measure the tension and activity of the aorta. Each blood vessel was measured at least 3 times and the average value was taken.

[0041] 2.7 Pathological staining

[0042] After the mice were killed, the aorta was isolated and part of the aorta tissue was used for pathological examination. Masson staining was used to evaluate the vascular morphology and collagen deposition.

[0043] 2.8 Statistical analysis SPSS25.0 software was used for statistical analysis. The data were expressed as mean ± standard deviation. One-way analysis of variance was used, and P < 0.05 was considered statistically significant.

[0044] 2.9 Results

[0045] The in vivo perfusion of angiotensin II (Ang II) by micro-osmotic pump to induce hypertension vascular remodeling model has the characteristics of simple operation, high success rate, easy replication, stability and reliability, and short modeling time. It can be widely used in the study of the pathogenesis of hypertension and the screening of antihypertensive drugs. In this study, the antihypertensive effect of the pharmaceutical composition of the present invention was verified by in vivo perfusion test of micro-osmotic pump, and the preliminary efficacy was explored. The experiment was designed to control group, model group (drinking pure water), and pharmaceutical composition group. According to the drug ratio, the prepared pharmaceutical composition liquid was given to mice to drink freely, and the amount of water consumed by mice was measured every 2 days. Compared with the control group, the serum IL-22 content of mice in the model group was significantly increased, and the difference was statistically significant (P < 0.05). After 28 days of Ang II perfusion, compared with the control group, the systolic and diastolic blood pressure of mice in the model group were significantly increased, and the difference was statistically significant (P < 0.05). The experimental results showed that the hypertension model was successfully established. See Table 1.

[0046] Table 1 Establishment of mouse hypertension model

[0047]

[0048] 2.3 Serological testing

[0049] Peripheral blood was collected 28 days after Ang II perfusion, and serum was collected after centrifugation to detect the levels of serum IL-1, IL-6, IL-22, TNF-α and TGF-β in mice. The results showed that the serum IL-1, IL-6, IL-22, TNF-α and TGF-β in the model group mice were significantly higher than those in the control group (P < 0.05), and the serum IL-1, IL-6, IL-22, TNF-α and TGF-β in the drug composition group mice were significantly lower than those in the model group (P < 0.05), and there was no significant difference compared with the control group, as shown in Table 2.

[0050] Table 2 Mouse serological tests

[0051]

[0052] 2.4 Comparison of mouse histopathology

[0053] Pathological staining showed that the aorta structure of mice in the control group and the drug combination group was intact, and no obvious pathological changes were observed; the expression of elastic fibers in mice in the model group increased, and collagen deposition around blood vessels was observed. Figure 1 .

[0054] 2.6 Flow cytometry detection of macrophage polarization

[0055] Compared with the control group, the expression of M1 macrophages (CD80 and CD86 markers) in the aorta of mice in the model group was significantly increased, and the expression of M2 macrophages (CD163 and CD206 markers) was decreased; the expression of M1 macrophages in the drug composition group was significantly decreased compared with the model group, and the expression of M2 macrophages was significantly increased, and there was no significant difference compared with the control group. Figure 2 .

[0056] 2.7 Caenorhabditis elegans SOD measurement

[0057] The experimental results showed that the survival rate of the model group (drinking pure water) was 10%, and the survival rate of the drug combination group was 100%; the survival rate was calculated as the total number of survival bars / total number of survival bars in the control group * 100%. The above results show that the oxidative damage model of Caenorhabditis elegans was successfully constructed, and the survival rate was significantly lower than that of the control group. The antioxidant damage effect of each drug administration group was significant and could significantly improve the survival rate of the oxidative damage model of Caenorhabditis elegans. The SOD content of each group was 62.5±12.3, 128.5±20.5, and 42.8±8.6, respectively. There was a very significant difference between the model group and the control group, indicating that the oxidative damage model of Caenorhabditis elegans was successfully established. The SOD content of each drug administration group was significantly higher than that of the model group, and showed a dose-effect relationship, indicating that the drug had a significant antioxidant effect, as shown in Table 3.

[0058] Table 3 Antioxidant test of Caenorhabditis elegans

[0059]

[0060] The experimental results show that the drug composition can reduce the systolic and diastolic blood pressure of hypertensive model mice, and at the same time reduce the SOD level of the Caenorhabditis elegans antioxidant test model Caenorhabditis elegans. It has the effect of inhibiting the secretion of chronic inflammatory factors and inhibiting the level of oxidative stress, thereby improving cardiovascular diseases and delaying the occurrence and development of hypertension.

Claims

1. A method for preparing a pharmaceutical composition, characterized in that: The pharmaceutical composition is prepared from the following raw materials in parts by weight: 40-60 parts of Prunella Vulgaris, 20-30 parts of chrysanthemum, 20-30 parts of cassia seeds and 20-30 parts of bitter buckwheat.

2. The method for preparing a pharmaceutical composition according to claim 1, characterized in that: The composition is prepared from the following raw materials in parts by weight: 60 parts of selfheal, 30 parts of chrysanthemum, 30 parts of cassia seed and 30 parts of bitter buckwheat.

3. A method for preparing a pharmaceutical composition according to claim 1 or 2, characterized in that: Relevant weight portions of herba prunellae, chrysanthemum, cassia seed and tartary buckwheat are weighed; supernatants of herba prunellae and chrysanthemum are obtained by traditional water decoction process; supernatants of cassia seed and tartary buckwheat are obtained by alcohol extraction process; the two parts of supernatants are concentrated, dried and mixed to obtain the pharmaceutical composition.

4. The method for preparing a pharmaceutical composition according to claim 2, characterized in that: The decoction process of Prunella vulgaris and chrysanthemum is as follows: 100g of medicinal materials are added with 6-8L of water, soaked for 10-12 hours, and then extracted at 90-100℃ for 1.5-2.5 hours; 50g of Cassia seed and 50g of Tartary Buckwheat were added to 6-8L of 95% ethanol, soaked for 4-6 hours, and then extracted at 80°C for 1.5-2.5 hours; The two parts of supernatant are concentrated, dried and mixed to obtain the pharmaceutical composition.

5. A pharmaceutical composition prepared by the method according to any one of claims 1 to 4.

6. Use of the pharmaceutical composition according to claim 5 in lowering blood pressure.

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