CN112603983A

CN112603983A - Pharmaceutical composition for treating COPD (chronic obstructive pulmonary disease) with pulmonary fibrosis

Info

Publication number: CN112603983A
Application number: CN202011639992.4A
Authority: CN
Inventors: 王成鑫; 杜娟; 吴玲敏; 王昀; 涂蕾; 阮景云; 唐春兰
Original assignee: Wuhan Kingmed Diagnostics Laboratory Co ltd
Current assignee: Wuhan Kingmed Diagnostics Laboratory Co ltd
Priority date: 2020-12-31
Filing date: 2020-12-31
Publication date: 2021-04-06

Abstract

The invention discloses a pharmaceutical composition for treating COPD (chronic obstructive pulmonary disease) with pulmonary fibrosis, which comprises the following components in parts by weight: 3-15 parts of astragalus membranaceus, 5-20 parts of salvia miltiorrhiza, 5-15 parts of scutellaria baicalensis, 3-10 parts of liquorice, 5-15 parts of safflower, 5-15 parts of angelica sinensis, 5-20 parts of spina date seed, 3-15 parts of ligusticum wallichii, 3-15 parts of schisandra chinensis and 5-20 parts of curcuma zedoary.

Description

Pharmaceutical composition for treating COPD (chronic obstructive pulmonary disease) with pulmonary fibrosis

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition for treating COPD (chronic obstructive pulmonary disease) with pulmonary fibrosis.

Background

Pulmonary interstitial fibrosis (PF) is a chronic pulmonary interstitial disease with the basic pathologies of pulmonary epithelial cell damage, inflammatory cell infiltration, fibrous tissue proliferation and finally pulmonary fibrosis leading to lung function impairment and respiratory failure. Many diseases can cause pulmonary fibrosis, such as Chronic Obstructive Pulmonary Disease (COPD) which is the most common disease of respiratory system and is about 80% accompanied by fibrosis pulmonary fibrosis, coronavirus infection such as former SARS, COVID-19 which is outbreak at present, anti-tumor agent, occupational dust, radiation and other factors. Pulmonary fibrosis may become the most serious complication after the epidemic situation passes, so that a treatment means for preventing pulmonary fibrosis in an early stage is imperative for the highly pathogenic coronavirus pneumonia, and the complications are reduced and the death rate is reduced through early intervention. Pulmonary fibrosis, in turn, can cause chronic pulmonary heart disease, lung cancer, etc., which, together with pulmonary fibrosis, are known as devastating lung diseases.

The etiology and pathogenesis of PF are complex, and the exact mechanism is not clear. So far, the traditional Chinese medicine composition still lacks powerful targeted medicine treatment for pulmonary fibrosis caused by various reasons, has poor treatment effect, and has 5-year survival rate of only 50 percent. With the intensive research on the pathogenesis of pulmonary interstitial fibrosis, early prevention and treatment are particularly important, and the problem is urgently needed to be solved. The conventional PF treatment method in modern medicine only can relieve the disease condition, and cannot achieve the satisfactory effect and cure purpose. At present, the hot spot and focus of domestic and international research are focused on searching pathogenic factors and pathogenic mechanisms of pulmonary fibrosis, and searching for effective methods which can inhibit inflammatory reaction and prevent fibrosis process, so that pulmonary fibrosis is possible to reverse and has low side effect. However, no effective treatment method exists in the current clinic, and the conventional clinical treatment method of the modern PF medicine only can relieve the disease condition, but cannot completely control the disease progress and even cannot achieve the aim of curing. Drugs such as glucocorticoids in combination with immunosuppressants, oxygen therapy and the like have been used for treatment, but these treatments have been found to have no significant effect on the eradication of such diseases. With lung failure, only lung organ transplantation can be performed, the effect is not satisfactory, and more importantly, the operation is expensive, the technical difficulty is high, and the supply of organs is difficult to find.

PF treatment by western medicine is difficult to achieve satisfactory effect, so the special effect of traditional Chinese medicine should be played. At present, the combination of Chinese and Western medicine respectively for treating pulmonary fibrosis has been advanced to a certain extent, but the documents are only rarely reported, and the medicaments for treating chronic obstructive pulmonary disease with pulmonary fibrosis and preventing and inhibiting lung cancer are not reported after the relevant documents are examined.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides a pharmaceutical composition for treating COPD (chronic obstructive pulmonary disease) with pulmonary fibrosis.

In order to achieve the purpose, the technical scheme provided by the invention is as follows:

the pharmaceutical composition for treating COPD with pulmonary fibrosis comprises the following components in parts by weight: 3-15 parts of astragalus membranaceus, 5-20 parts of salvia miltiorrhiza, 5-15 parts of scutellaria baicalensis, 3-10 parts of liquorice, 5-15 parts of safflower, 5-15 parts of angelica sinensis, 5-20 parts of spina date seed, 3-15 parts of ligusticum wallichii, 3-15 parts of schisandra chinensis and 5-20 parts of curcuma zedoary.

Preferably, the pharmaceutical composition comprises the following components in parts by weight: 3 parts of astragalus membranaceus, 5 parts of salvia miltiorrhiza, 5 parts of scutellaria baicalensis, 3 parts of liquorice, 5 parts of safflower, 5 parts of angelica sinensis, 5 parts of spina date seed, 3 parts of ligusticum wallichii, 3 parts of schisandra chinensis and 5 parts of curcuma zedoary.

Preferably, the pharmaceutical composition comprises the following components in parts by weight: 15 parts of astragalus membranaceus, 20 parts of salvia miltiorrhiza, 15 parts of scutellaria baicalensis, 10 parts of liquorice, 15 parts of safflower, 15 parts of angelica sinensis, 20 parts of spina date seed, 15 parts of ligusticum wallichii, 15 parts of schisandra chinensis and 20 parts of curcuma zedoary.

Preferably, the pharmaceutical composition comprises the following components in parts by weight: 3 parts of astragalus membranaceus, 20 parts of salvia miltiorrhiza, 5 parts of scutellaria baicalensis, 10 parts of liquorice, 5 parts of safflower, 15 parts of angelica sinensis, 5 parts of spina date seed, 15 parts of ligusticum wallichii, 3 parts of schisandra chinensis and 20 parts of curcuma zedoary.

Preferably, the pharmaceutical composition comprises the following components in parts by weight: 15 parts of astragalus membranaceus, 5 parts of salvia miltiorrhiza, 15 parts of scutellaria baicalensis, 3 parts of liquorice, 15 parts of safflower, 5 parts of angelica sinensis, 20 parts of spina date seed, 3 parts of ligusticum wallichii, 15 parts of schisandra chinensis and 5 parts of curcuma zedoary.

Preferably, the medicinal composition is prepared into an extract by a water boiling and alcohol precipitation method.

More preferably, the crude drug content in the extract is 10 g/mL.

The pharmaceutical composition provided by the invention has the function of poisoning A549 lung cancer cells by treating COPD (chronic obstructive pulmonary disease) accompanied with pulmonary fibrosis, and can be applied as a poisoning agent for the A549 lung cancer cells.

The invention optimally screens out the pharmaceutical composition for treating pulmonary fibrosis according to the principles of promoting blood circulation to remove blood stasis, breaking blood and resolving masses. The prescription is prepared from: 3-15g of astragalus root, 5-20g of salvia miltiorrhiza, 5-15g of scutellaria root, 3-10g of liquorice, 5-15g of safflower, 5-15g of angelica, 5-20g of Chinese violet, 3-15g of szechuan lovage rhizome, 3-15g of Chinese magnoliavine fruit, 5-20g of zedoary and the like. Radix astragali and angelica sinensis have the functions of tonifying qi, activating blood and nourishing blood, radix salviae miltiorrhizae, safflower, rhizoma ligustici wallichii and rhizoma zedoariae have the functions of activating blood, dissolving stasis and promoting qi circulation, spina date seed has the functions of clearing heat, promoting diuresis and resolving turbidity, and liquorice is used for harmonizing the effects of the other drugs. The method for improving and relieving the pulmonary fibrosis by applying the GHLF is an initial research.

In a word, the pharmaceutical composition has a good effect on treating pulmonary fibrosis, can effectively treat and improve COPD (chronic obstructive pulmonary disease) with pulmonary fibrosis, obviously improves clinical symptoms of patients, can effectively treat and improve COPD with pulmonary fibrosis, obviously improves clinical symptoms of patients, and enhances exercise endurance and cardiopulmonary function of patients, and has a good effect. Can improve the lymphocyte transformation rate in a test tube, has obvious poisoning effect on A549 lung cancer cells, has in vitro antibacterial and anti-inflammatory effects, provides a solid theoretical basis for the traditional Chinese medicine to treat pulmonary fibrosis, and provides a theoretical basis for wider application in clinic. The invention has real and reliable data and standard operation, meets the research ethical requirements of people and the relevant regulations of the national health council of students.

Drawings

FIG. 1 is a CT slice before treatment;

FIG. 2 is a CT slice after treatment;

FIG. 3: larger in the left panel are transitional lymphocytes: slightly larger than mature lymphocytes; small are mature lymphocytes: and the size of the cells which are not transformed is consistent with that of the uncultured lymphocytes in peripheral blood; larger reticulocyte-like lymphocytes in the right panel: the cells are obviously enlarged and are multiple times of mature untransformed cells, the shapes of the cells are irregular, the cells are approximately amoebic-like, and nucleoli can be seen.

Detailed Description

An experimental method for the obvious poisoning effect of the pharmaceutical composition on A549 lung cancer cells comprises the following steps:

materials and methods

1. Material

The preparation method of the pharmaceutical composition comprises selecting the herbs of the above components (radix astragali 3 parts, Saviae Miltiorrhizae radix 5 parts, Scutellariae radix 5 parts, Glycyrrhrizae radix 3 parts, Carthami flos 5 parts, radix Angelicae sinensis 5 parts, herba Violae 5 parts, rhizoma Ligustici Chuanxiong 3 parts, fructus Schisandrae 3 parts, Curcumae rhizoma 5 parts), decocting with water, and precipitating with ethanol to obtain 1:10 extract; the lung cancer A549 cell strain is provided by friendly people.

2. Method of producing a composite material

The lung cancer A549 cells were cultured in DMEM complete medium containing lO% Fetal Bovine Serum (FBS) and formulated to 4X lO according to the method described in the literature⁵Cell suspension/ml, seeded in 96-well plates, tumor cell suspension l00ml per well. Adding the extracts with different concentrations (5 groups with different concentrations of 1g, 0.4g, 0.16g, 0.064g, and 0.0256g/ml for each group, and using test tube without Chinese medicinal materials as blank control group, and repeating four same samples for each concentration). lOO μ l of medicated solution is added into each well; the blank control group was added with an equal volume of DMEM medium containing 8% FBS (8 identical samples were repeated), incubated at 37 ℃ in a 5% C02 incubator for 48 hours, aspirated to remove the supernatant, washed with PBS 2 times, added with 12. mu.l of MTT stock (10mg/m1), and placed at 37 ℃ in a 5% C0 medium₂The culture is continued for 4 hours under the condition, the supernatant is removed, PBS is washed once, lOO mu l of dimethyl sulfoxide (DMSO) is added to dissolve blue-purple formazan particles generated by MTT reduction, and an enzyme linked immunosorbent assay detector (DG 3022A) is used for determining the OD value. The measurement wavelength was 490nm (effective reading between 0.01-1. O). The influence of the medicines on the MTT OD value of the A549 cancer cells and the killing rate of the medicines on the tumor cells are calculated according to the following formula:

3. as a result:

through calculation, the medicament with each group concentration has obvious poisoning effect on A549 lung cancer cells and can cause cancer cell death, and the higher the medicament concentration is, the higher the killing rate on tumor cells is, namely, the stronger the poisoning effect on the cells is.

Secondly, the experimental method for the lymphocyte transformation rate of the pharmaceutical composition comprises the following steps:

the influence of the micro whole blood cell culture method on the human lymphocyte transformation rate is observed through in vitro experiments, and the effect of the prescription on the human immunologic function is comprehensively evaluated.

Materials and methods

1. Material

The preparation method of the pharmaceutical composition comprises selecting the above herbs (radix astragali 3, Saviae Miltiorrhizae radix 5, Scutellariae radix 5, Glycyrrhrizae radix 3, Carthami flos 5, radix Angelicae sinensis 5, spina Gleditsiae 5, rhizoma Ligustici Chuanxiong 3, fructus Schisandrae 3, and Curcumae rhizoma 5), decocting with water, and precipitating with ethanol to obtain 1:10 extract. Complete culture of RPMI1640 containing Phytohemagglutinin (PHA) was purchased from Haires bioengineering, Qingdao (PHA concentration 0.3 mg/ml. whole blood: from an adult who had not taken any drug for one three months.

2. Method of producing a composite material

2.1 culturing isolated lymphocytes

The effect of the drug composition on lymphocyte transformation rate according to the method described in the literature, 0.2ml of drug solutions containing various concentrations of traditional Chinese medicines are added into test tubes containing 0.3ml of whole blood, 5 drug concentration groups are provided, the concentration of each group is 1g, 0.4g, 0.16g, 0.064g and 0.0256g/ml respectively, and the test tubes without traditional Chinese medicines are used as a control group. 2.5ml of prepared 1640 complete culture solution is added into each test tube, and two same samples are respectively repeated at various concentrations. Incubating for 72 hours in a constant-temperature incubator at 37 ℃, taking out, centrifuging for 10 minutes at 1500 rpm, removing supernatant, adding 8ml of pre-warmed 0 and 75M potassium chloride solution, repeatedly blowing and beating by using a pipette to fully dissolve erythrocytes, placing in a water bath incubator at 37 ℃ for 30 minutes, adding 1ml of fixing agent (methanol: glacial acetic acid: 3:1), uniformly mixing, centrifuging under the same condition to remove supernatant, adding 8ml of fixing agent, and repeating the previous operation to obtain pure lymphocytes. The normal adult lymphocyte transformation rates in the literature are referred to as normal control values for statistical treatment.

2.2 preparation of smear specimen and microscopic examination

After the culture is finished, sucking lymphocyte fluid smears, drying, smearing two smears of each sample by using a Wright-Giemsa method, counting by microscopic examination of doctors without knowing the concentration of each group of medicine, counting 100 lymphocytes of each smear according to the head, the body and the tail respectively, and obtaining the average value as the final lymphocyte conversion rate.

The lymphocyte transformation rate was calculated as follows:

2.3 statistical methods

Statistical treatment by T-test

Results

After the lymphocytes are cultured by adding mitogens, the lymphocytes can be transformed into mother cells with various forms, and the mastery of the morphological characteristics is the key for determining the transformation of the lymphocytes, and the lymphocytes are generally divided into the following types of cells (as shown in figure 3):

1. reticulocyte-like lymphocytes: the cells are obviously enlarged and are multiple times of mature untransformed cells, the shapes of the cells are irregular, the cells are approximately amoebic, and the nucleolus is obvious.

2. Lymphoblast cell: is 3-4 times larger than mature lymphocyte, cytoplasm sometimes extends out of pseudopodia or has vacuole, and nucleoplasm is loose and has nucleolus.

3. Transitional lymphocytes: slightly larger than mature lymphocytes and distinguished from mature lymphocytes by nucleoli.

4. Mature lymphocytes: also, the size of the untransformed cells was consistent with that of uncultured lymphocytes in peripheral blood, and the cells had large nuclei, were rounded, homogeneous, and generally did not show cytoplasm.

The transformed lymphocytes refer to lymphoblasts, reticulocytes, and transitional lymphoid cells, and the specific results are shown in table 1:

FIG. 1 Effect of drugs for treating COPD with pulmonary fibrosis on lymphocyte transformation ratio

As shown in Table 1, the traditional Chinese medicine can improve the conversion rate of lymphocytes, namely the immunologic function, compared with a blank control group in the concentration range of 0.4-0.0256 g/ml, but has no statistical significance (P is more than 0.05) in the 0.4g/ml group, and has no statistical significance in the 0.4g/ml group

The group of 0.16-0.025 g/ml can obviously improve the conversion rate of lymphocytes, has significant difference (the P values are respectively less than 0.05 and less than 0.01), and shows a certain dose-effect relationship. As shown in FIG. 1, the effect on lymphocyte transformation efficiency increases with decreasing concentration.

The traditional Chinese medicine preparation has good effect of treating Chronic Obstructive Pulmonary Disease (COPD) with Pulmonary fibrosis. The optimal drug concentration and the optimal dose of a patient are reserved, the drug is taken for ten days in one course of treatment, after five courses of treatment, the results of objective curative effect indexes, namely six-minute walking test, dyspnea index, pulmonary function, imaging and other indexes, show that the curative effect of a treatment group is obviously superior to that of a control group, the clinical symptoms of the patient are obviously improved, the exercise endurance and the cardiopulmonary function of the patient are enhanced, even the bad mood of the patient is improved, the total effective rate of a Chinese and western medicine combination group is higher than that of a western medicine treatment group, and the life quality of the patient is improved. The imaging shows that the lung fibrosis is effectively treated, improved and ablated (as shown in figures 1-2), figure 1 shows that the CT film before treatment has higher density images of spot-shaped, grid-shaped and cord-shaped fibrosis in the solid-variable area, and figure 2 shows that the symptoms of the lung fibrosis are gradually reduced or even completely disappear after the treatment of the traditional Chinese medicine.

The pharmaceutical composition has the effects of treating COPD, resisting inflammation, resisting fibrosis, resisting tumor and improving immune function, so that the pharmaceutical composition can be a new medicine for assisting COPD with PF treatment, especially the traditional Chinese medicine for treating pulmonary fibrosis has few documents, and the medicine for preventing and inhibiting lung cancer and improving immune function is not reported.

The project applies high and new technologies such as biology, cytology, immunology and the like to comprehensively research the prescription, partially clarifies the treatment effect, action mechanism, action part and medicinal effect of the medicinal composition on pulmonary fibrosis, lung cancer inhibition and prevention and the like, provides scientific medication basis for clinic and lays scientific foundation for developing the international market of traditional Chinese medicines.

The invention aims to provide a new treatment way for lung injury, fibrosis and prevention of subsequent lung cancer, which contributes to the application of the Chinese medicament in the treatment of lung fibrosis.

Claims

1. The pharmaceutical composition for treating COPD (chronic obstructive pulmonary disease) with pulmonary fibrosis is characterized by comprising the following components in parts by weight: 3-15 parts of astragalus membranaceus, 5-20 parts of salvia miltiorrhiza, 5-15 parts of scutellaria baicalensis, 3-10 parts of liquorice, 5-15 parts of safflower, 5-15 parts of angelica sinensis, 5-20 parts of spina date seed, 3-15 parts of ligusticum wallichii, 3-15 parts of schisandra chinensis and 5-20 parts of curcuma zedoary.

2. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises the following components in parts by weight: 3 parts of astragalus membranaceus, 5 parts of salvia miltiorrhiza, 5 parts of scutellaria baicalensis, 3 parts of liquorice, 5 parts of safflower, 5 parts of angelica sinensis, 5 parts of spina date seed, 3 parts of ligusticum wallichii, 3 parts of schisandra chinensis and 5 parts of curcuma zedoary.

3. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises the following components in parts by weight: 15 parts of astragalus membranaceus, 20 parts of salvia miltiorrhiza, 15 parts of scutellaria baicalensis, 10 parts of liquorice, 15 parts of safflower, 15 parts of angelica sinensis, 20 parts of spina date seed, 15 parts of ligusticum wallichii, 15 parts of schisandra chinensis and 20 parts of curcuma zedoary.

4. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises the following components in parts by weight: 3 parts of astragalus membranaceus, 20 parts of salvia miltiorrhiza, 5 parts of scutellaria baicalensis, 10 parts of liquorice, 5 parts of safflower, 15 parts of angelica sinensis, 5 parts of spina date seed, 15 parts of ligusticum wallichii, 3 parts of schisandra chinensis and 20 parts of curcuma zedoary.

5. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition comprises the following components in parts by weight: 15 parts of astragalus membranaceus, 5 parts of salvia miltiorrhiza, 15 parts of scutellaria baicalensis, 3 parts of liquorice, 15 parts of safflower, 5 parts of angelica sinensis, 20 parts of spina date seed, 3 parts of ligusticum wallichii, 15 parts of schisandra chinensis and 5 parts of curcuma zedoary.

6. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is prepared into an extract by a water decoction and alcohol precipitation method.

7. The pharmaceutical composition of claim 6, wherein the extract contains crude drug at a concentration of 10 g/mL.

8. The use of the pharmaceutical composition of claim 1 in the preparation of an a549 lung cancer cell toxicant.