A traditional Chinese medicine composition, a preparation method and application thereof

Abstract

This invention provides a traditional Chinese medicine composition for treating and / or preventing lung inflammation-related diseases, wherein the herbal extract contains three active components: Scutellaria baicalensis extract, Stephania tetrandra extract, and Glycyrrhiza uralensis extract. This traditional Chinese medicine composition can be used to treat acute lung injury caused by various factors; currently, there is a lack of drugs for this disease in clinical practice, and this composition can fill that gap.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a traditional Chinese medicine composition, its preparation method, and its uses. Background Technology

[0002] Acute lung injury (ALI) is a critical illness caused by the infiltration and activation of inflammatory cells in lung tissue by various pathogenic factors inside and outside the lungs, which release a large number of inflammatory factors, forming a cytokine storm, causing damage to alveolar epithelial cells and vascular endothelial cells, leading to pulmonary edema and impaired pulmonary ventilation / gas exchange.

[0003] Currently, there are no effective drugs or specific treatments for ALI in clinical practice. The main treatment is glucocorticoids combined with mechanical ventilation. Although this can relieve the inflammatory response and improve pulmonary ventilation dysfunction, it can cause adverse reactions such as immunosuppression, coagulation dysfunction and osteoporosis. Therefore, its clinical use is very limited.

[0004] Respiratory infectious diseases spread rapidly and are difficult to prevent, resulting in a high incidence of ALI, which seriously threatens human life and health. Therefore, it is urgent to find an effective and safe drug to treat ALI. Summary of the Invention

[0005] To address the shortcomings of existing technologies, one objective of this invention is to provide a traditional Chinese medicine composition for treating and / or preventing lung inflammation-related diseases. The composition comprises three effective components: Scutellaria baicalensis extract, Stephania tetrandra extract, and Glycyrrhiza uralensis extract. This traditional Chinese medicine composition can be used to treat acute lung injury caused by various factors, a condition currently lacking in clinical practice; this composition fills that gap.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows:

[0007] A traditional Chinese medicine composition, made from the following raw materials: Scutellaria baicalensis, Stephania tetrandra, and Glycyrrhiza uralensis, wherein, by weight,

[0008] Scutellaria baicalensis, 5-30 parts by weight;

[0009] 5-30 parts by weight of Stephania tetrandra;

[0010] Licorice 1-25 parts by weight.

[0011] According to the traditional Chinese medicine composition of the present invention, wherein, by weight,

[0012] Scutellaria baicalensis, 5-20 parts by weight;

[0013] 5-20 parts by weight of Stephania tetrandra;

[0014] Licorice 5-20 parts by weight.

[0015] According to the traditional Chinese medicine composition of the present invention, wherein, by weight,

[0016] Scutellaria baicalensis 10-20 parts by weight;

[0017] 10-20 parts by weight of Stephania tetrandra;

[0018] Licorice 5-15 parts by weight.

[0019] The second objective of this invention is to provide a method for preparing the above-mentioned traditional Chinese medicine composition.

[0020] The preparation method of the traditional Chinese medicine composition of the present invention is achieved through the following technical solution:

[0021] A method for preparing the above-mentioned traditional Chinese medicine composition includes the following steps:

[0022] Weigh out Scutellaria baicalensis according to the prescription ratio, extract with water under reflux, cool, filter, concentrate the filtrate under reduced pressure, and freeze dry to obtain crude Scutellaria baicalensis extract; add the crude Scutellaria baicalensis extract to macroporous adsorption resin, elute with water and ethanol in sequence, concentrate the eluent under reduced pressure, and freeze dry to obtain Scutellaria baicalensis extract (total flavonoids of Scutellaria baicalensis).

[0023] Weigh out the ingredients of Stephania tetrandra according to the prescription ratio, extract with ethanol under reflux, cool, filter, concentrate the filtrate under reduced pressure, and freeze dry to obtain the crude extract of Stephania tetrandra; add the crude extract of Stephania tetrandra to a macroporous adsorption resin, elute with water and ethanol in sequence, concentrate the eluent under reduced pressure, and freeze dry to obtain the Stephania tetrandra extract (total alkaloids of Stephania tetrandra).

[0024] Licorice was weighed according to the prescription ratio, extracted by reflux with ethanol, cooled, filtered, concentrated under reduced pressure, and freeze-dried to obtain crude licorice extract; the crude licorice extract was added to macroporous adsorption resin, eluted with water and ethanol in sequence, concentrated under reduced pressure, and freeze-dried to obtain licorice extract (total triterpenoids of licorice).

[0025] The above-mentioned Scutellaria baicalensis extract, Stephania tetrandra extract and Glycyrrhiza uralensis extract are mixed to obtain the traditional Chinese medicine composition.

[0026] According to the preparation method of the present invention, the preparation method includes the following steps:

[0027] Weigh out Scutellaria baicalensis according to the prescription ratio, and extract three times by reflux with 10-14 times (preferably 12 times) of water. The first extraction lasts for 1-3 hours (preferably 2 hours), the second for 1-2 hours (preferably 1.5 hours), and the third for 0.8-1.5 hours (preferably 1 hour). Cool, filter, and concentrate the filtrate under reduced pressure at 70-90℃ (preferably 80℃). Freeze-dry to obtain crude Scutellaria baicalensis extract. Accurately weigh the above crude Scutellaria baicalensis extract, add it to macroporous adsorption resin, and elute sequentially with water and 50-90% ethanol. Discard the water eluent, concentrate the ethanol eluent under reduced pressure, and freeze-dry to obtain Scutellaria baicalensis extract.

[0028] Weigh the ingredients of Stephania tetrandra according to the prescription ratio, and extract three times with 8-12 (preferably 10 times) times the amount of 60-90% ethanol under reflux. The first extraction lasts for 1-3 hours (preferably 2 hours), the second for 1-2 hours (preferably 1.5 hours), and the third for 1-2 hours (preferably 1.5 hours). Cool, filter, and concentrate the filtrate under reduced pressure at 70-90℃ (preferably 80℃). Freeze-dry to obtain the crude extract of Stephania tetrandra. Accurately weigh the above crude extract of Stephania tetrandra, add it to a macroporous adsorption resin, and elute sequentially with water and 50-90% ethanol. Discard the water eluent, concentrate the ethanol eluent under reduced pressure, and freeze-dry to obtain the Stephania tetrandra extract.

[0029] Licorice was weighed according to the prescription ratio and extracted three times by reflux with 8-12 (preferably 10 times) times the amount of 60-90% ethanol. The first extraction lasted 1-3 hours (preferably 2 hours), the second extraction lasted 1-2 hours (preferably 1.5 hours), and the third extraction lasted 1-2 hours (preferably 1.5 hours). After cooling, the extract was filtered, and the filtrate was concentrated under reduced pressure at 70-90℃ (preferably 80℃) and freeze-dried to obtain crude licorice extract. The crude licorice extract was accurately weighed and added to a macroporous adsorption resin. The extract was eluted sequentially with water and 50-90% ethanol. The water eluent was discarded, and the ethanol eluent was concentrated under reduced pressure and freeze-dried to obtain licorice extract.

[0030] The above-mentioned Scutellaria baicalensis extract, Stephania tetrandra extract and Glycyrrhiza uralensis extract are mixed to obtain the traditional Chinese medicine composition.

[0031] According to the preparation method of the present invention, in the preparation step of Scutellaria baicalensis extract, the macroporous adsorption resin used is selected from one or more of D101, HPD-300, AB-8, HPD-600 and HPD-826; the mass ratio (g / g) of Scutellaria baicalensis to macroporous adsorption resin is 1:2 to 1:4, preferably 1:3.

[0032] According to the preparation method of the present invention, in the preparation step of Stephania tetrandra extract, the macroporous adsorption resin used is selected from one or more of D101, HPD-300, AB-8, HPD-600 and HPD-826; the mass ratio (g / g) of Stephania tetrandra to macroporous adsorption resin is 1:0.5 to 1:0.8, preferably 1:0.6.

[0033] According to the preparation method of the present invention, in the preparation step of licorice extract, the macroporous adsorption resin used is selected from one or more of D101, HPD-300, AB-8, HPD-600 and HPD-826; the mass ratio (g / g) of licorice to macroporous adsorption resin is 1:1 to 1:2, preferably 1:1.5.

[0034] A third objective of this invention is to provide the use of the above-mentioned traditional Chinese medicine composition in the preparation of a medicament for treating and / or preventing lung inflammation-related diseases.

[0035] According to the uses described in this invention, the lung inflammation-related diseases include, but are not limited to, lung inflammation, acute lung injury, and pulmonary fibrosis.

[0036] A fourth objective of this invention is to provide a pharmaceutical formulation comprising the aforementioned traditional Chinese medicine composition and a pharmaceutically acceptable carrier. Generally, a "pharmaceutically acceptable carrier" refers to a medium generally acceptable in the art for delivering bioactive substances to cells or for delivering bioactive substances to animals (specifically mammals), including adjuvants, excipients, or solvents, such as diluents, preservatives, fillers, flow modifiers, penetration enhancers, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, lubricants, and dispersants. The choice of a pharmaceutically acceptable carrier depends on the administration route and the nature of the dosage form.

[0037] According to the pharmaceutical formulation of the present invention, the pharmaceutical formulation is administered orally, intravenously, locally, by inhalation or spray, sublingually, transdermally, or rectally, or by other means.

[0038] According to the pharmaceutical formulations of the present invention, the pharmaceutical formulation may be tablets, granules, hard capsules, mixtures, pills, drop pills, effervescent tablets, injections, powder injections, or soft capsules. The various dosage forms of pharmaceutical formulations of the present invention can be prepared using conventional methods in the art.

[0039] It should be noted that the carriers, administration methods, and dosage forms that can be added to the traditional Chinese medicine composition of the present invention are not limited by the above-mentioned content.

[0040] The "pulmonary inflammation-related diseases" described in this invention refer to pneumonia, acute lung injury, and lung diseases with pathology consistent with pneumonia and / or acute lung injury caused by various factors. "Pneumonia and acute lung injury caused by various factors" as described in this invention refers to lung diseases with pathology consistent with pneumonia and / or acute lung injury caused by infectious factors (viruses or bacteria) and non-infectious factors (foreign body aspiration), but is not limited to these.

[0041] Beneficial effects

[0042] Compared with the prior art, the advantages of the present invention are as follows:

[0043] 1. The traditional Chinese medicine composition of the present invention is used to treat acute lung injury (ALI) caused by various factors. Currently, there is a lack of drugs for this disease in clinical practice, and the traditional Chinese medicine composition of the present invention can fill this gap.

[0044] 2. The traditional Chinese medicine composition of this invention is based on a combination of multiple effective components of traditional Chinese medicine, which can exert effects on multiple targets and at multiple levels. The combination of multiple components can overcome the limited efficacy, adverse reactions, and drug resistance of single-component treatments. Animal experimental studies have shown that the traditional Chinese medicine composition of this invention significantly improves the pathological damage of LPS-induced ALI and has a significant inhibitory effect on pulmonary edema, which can effectively curb clinical deaths caused by respiratory failure due to pulmonary edema.

[0045] 3. The components and their combinations contained in the traditional Chinese medicine composition of the present invention have been proven by numerous experimental studies to have no toxic side effects within the effective dosage range, which can ensure the safety of clinical medication. Attached Figure Description

[0046] Figure 1 The image shows a pathological section of lung tissue from a mouse with LPS-induced ALI, observed under a 40X light microscope.

[0047] Figure 2 The image shows a pathological section of lung tissue from a mouse with LPS-induced ALI, observed under a 100X light microscope. Detailed Implementation

[0048] Preferred embodiments of the invention will now be described in more detail. While preferred embodiments of the invention are described below, it should be understood that the invention can be implemented in various forms and should not be limited to the embodiments set forth herein.

[0049] The described embodiments are merely some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention are within the scope of protection of the present invention.

[0050] Information on the materials used in the embodiments and experimental examples of this invention is as follows:

[0051] Scutellaria baicalensis slices, Stephania tetrandra slices, and Glycyrrhiza uralensis slices were purchased from the Pharmacy Department of Xiyuan Hospital, China Academy of Chinese Medical Sciences.

[0052] Unless otherwise specified, all materials used in the following examples and experimental cases are commercially available.

[0053] Preparation of the pharmaceutical composition in the examples

[0054] The traditional Chinese medicine composition of the present invention is prepared by weighing each raw material according to the prescription ratio in Table 1 below.

[0055] Table 1

[0056]

[0057] The traditional Chinese medicine compositions in each embodiment of the present invention are prepared according to the following method, including the following steps:

[0058] Preparation of Scutellaria baicalensis extract:

[0059] Accurately weigh the Scutellaria baicalensis slices and extract them three times with 12 times their volume of water under reflux: 2 hours for the first extraction, 1.5 hours for the second, and 1 hour for the third. Cool and filter the extract, concentrate the filtrate under reduced pressure at 80°C, and freeze-dry to obtain crude Scutellaria baicalensis extract. Accurately weigh the crude Scutellaria baicalensis extract and add it to pretreated AB-8 macroporous adsorption resin. Pass the extract through the macroporous adsorption resin at a ratio of crude drug to macroporous resin of 1:3. Discard the water eluent and concentrate the 70% ethanol eluent under reduced pressure. Freeze-dry to obtain Scutellaria baicalensis extract (total flavonoids from Scutellaria baicalensis), with an extraction rate of 25-35%.

[0060] Preparation of Stephania tetrandra extract:

[0061] Accurately weigh the sliced ​​Stephania tetrandra, and extract three times with 10 times the volume of 70% ethanol under reflux: 2 hours for the first extraction, 1.5 hours for the second, and 1.5 hours for the third. Cool, filter, and concentrate the filtrate under reduced pressure at 80°C. Freeze-dry to obtain the crude extract of Stephania tetrandra. Accurately weigh the above crude extract of Stephania tetrandra, add it to pretreated HPD-300 macroporous adsorption resin, and pass the extract through the macroporous adsorption resin at a ratio of 1:0.6 (crude drug to macroporous resin). Discard the water washings, concentrate the 80% ethanol eluent under reduced pressure, and freeze-dry to obtain the Stephania tetrandra extract (total alkaloids of Stephania tetrandra), with an extraction rate of 3-5%.

[0062] Preparation of licorice extract:

[0063] Accurately weigh licorice slices and extract them three times with 10 times the volume of 70% ethanol under reflux: 2 hours for the first extraction, 1.5 hours for the second, and 1.5 hours for the third. Cool and filter the extract, concentrate the filtrate under reduced pressure at 80°C, and freeze-dry to obtain crude licorice extract. Accurately weigh the crude licorice extract and add it to pretreated AB-8 macroporous adsorption resin. Pass the extract through the macroporous adsorption resin at a ratio of 1:1.5 (crude drug to macroporous resin). Discard the water washings and concentrate the 60% ethanol eluent under reduced pressure. Freeze-dry to obtain licorice extract (total triterpenoids of licorice), with an extraction rate of 9-14%.

[0064] The above-mentioned Scutellaria baicalensis extract, Stephania tetrandra extract and Glycyrrhiza uralensis extract are mixed to obtain the traditional Chinese medicine composition.

[0065] Experimental Section

[0066] Experimental Example 1

[0067] 1. Experimental Materials

[0068] 1.1 Animals

[0069] Eighty-one male SPF-grade Balb / c mice, weighing 18–20 g, were purchased from SPF (Beijing) Biotechnology Co., Ltd. and housed at the Animal Experiment Center of Xiyuan Hospital, China Academy of Chinese Medical Sciences, in an environment with a temperature of 22±2℃ and a relative humidity of 55%±2%, with 12 hours of alternating light and dark conditions, and free access to food and water.

[0070] 1.2 Drugs and Reagents

[0071] The Scutellaria baicalensis extract, Stephania tetrandra extract, and Glycyrrhiza uralensis extract in the traditional Chinese medicine composition of the present invention were all prepared by the Institute of Basic Research, Xiyuan Hospital, China Academy of Chinese Medical Sciences.

[0072] Lipopolysaccharide (LPS) O55: B55, a product of Sigma, USA, batch number: 0000223034.

[0073] 2. Experimental Methods

[0074] 2.1 Experimental Design Scheme

[0075] Referring to the dosage guidelines in the 2020 edition of the Chinese Pharmacopoeia (Part I), the clinical crude drug dosages for Scutellaria baicalensis (P314), Stephania tetrandra (P155), and Glycyrrhiza uralensis (P88) were all set at 10g crude drug / person / day. Based on the average extraction rates of Scutellaria baicalensis extract, Stephania tetrandra extract, and Glycyrrhiza uralensis extract being 30.8%, 4.2%, and 11.3%, respectively, and the clinical average body weight being 70kg / person, and the dose coefficient between mice and humans being 12.3, the mouse equivalent dose of each extract was calculated as follows: A. Scutellaria baicalensis extract 540mg / kg, B. Stephania tetrandra extract 75mg / kg, C. Glycyrrhiza uralensis extract 200mg / kg. This experimental example compares the effects of single extracts, combinations of extracts, and combinations of three extracts on ALI.

[0076] 2.2 Grouping and Administration Methods

[0077] After three days of acclimatization feeding, 81 Balb / c mice were randomly divided into 9 groups according to the randomization principle: normal control group, model group, group A, group B, group C, group A+B, group A+C, group B+C, and group A+B+C, with 9 mice in each group. The traditional Chinese medicine composition was prepared as an aqueous solution according to the dosage designed in the experimental design and administered by gavage at a dose of 0.1 mL / 10 g BW once daily for 7 consecutive days. The normal control group and the model group were given an equal volume of distilled water.

[0078] 2.3 ALI Modeling Method

[0079] On the seventh day after gavage administration, mice were immediately anesthetized with isoflurane and ALI modeled by nasal inhalation of LPS solution at a dose of 1 mg / kg. The normal control group inhaled an equal amount of physiological saline through the nasal cavity.

[0080] 2.4 Indicator Testing

[0081] Twenty-four hours after the ALI model was established using LPS, mice were anesthetized with 0.5% sodium pentobarbital, euthanized by exsanguination through the abdominal aorta, and the lungs were dissected. The wet weight (W) of the right lung was measured, and the dry weight (D) was measured after placing it in a 60°C oven for 24 hours. The edema index (W / D) was calculated. The left lung was repeatedly irrigated three times with 0.8 mL of pre-cooled PBS, and the bronchoalveolar lavage fluid (BALF) was collected.

[0082] 3. Statistics and Analysis

[0083] Experimental data results are expressed as "mean ± standard deviation". This indicates that a one-way ANOVA was performed using SPSS 20.0 software, and a p-value < 0.05 was considered statistically significant.

[0084] 4. Results

[0085] The effects of the drugs on LPS-induced ALI in mice are shown in Table 2. Compared with the normal control group, the edema index, white blood cell count and protein content in bronchoalveolar lavage fluid (BALF) of the model group were significantly increased (P<0.05 or P<0.01), indicating significant lung tissue damage. Compared with the model group, each drug administration group could reduce the edema index, white blood cell count and protein content in bronchoalveolar lavage fluid (BALF) to varying degrees. The combination A+B+C group had the most significant effect (all P<0.01), which was better than the effects of single extract and the combination of two extracts.

[0086] Table 2. Effects of the drug on LPS-induced ALI in mice (n=9)

[0087]

[0088] Note: A is Scutellaria baicalensis extract, B is Stephania tetrandra extract, and C is Glycyrrhiza uralensis extract. Compared with the model group, *P<0.05 and **P<0.01.

[0089] 5. Conclusion

[0090] Based on the experimental results of Example 1, the combination of Scutellaria baicalensis, Stephania tetrandra, and Glycyrrhiza uralensis extracts of the present invention, when administered prophylactically, can significantly improve acute lung injury caused by LPS, with better effects than single extracts and combinations of two extracts.

[0091] Experiment Example 2: Prophylactic Drug Administration Experiment

[0092] 1. Experimental Materials

[0093] 1.1 Animals

[0094] Forty-five male SPF-grade Balb / c mice, weighing 18–20 g, were purchased from SPF (Beijing) Biotechnology Co., Ltd. and housed at the Animal Experiment Center of Xiyuan Hospital, China Academy of Chinese Medical Sciences, in an environment with a temperature of 22±2℃ and a relative humidity of 55%±2%, with 12 hours of alternating light and dark conditions. They had free access to food and water.

[0095] 1.2 Drugs and Reagents

[0096] The Scutellaria baicalensis extract, Stephania tetrandra extract, and Glycyrrhiza uralensis extract in the traditional Chinese medicine composition of the present invention were all prepared by the Institute of Basic Research, Xiyuan Hospital, China Academy of Chinese Medical Sciences.

[0097] Lipopolysaccharide (LPS) O55: B55, a product of Sigma, USA, batch number: 0000223034.

[0098] 2. Experimental Methods

[0099] 2.1 Experimental Design Scheme

[0100] Referring to the dosage guidelines in the 2020 edition of the Chinese Pharmacopoeia (Part I), the clinical crude drug dosages for Scutellaria baicalensis (P314), Stephania tetrandra (P155), and Glycyrrhiza uralensis (P88) were set at 5, 10, and 20 g crude drug / person / day. Based on the average extraction rates of Scutellaria baicalensis extract, Stephania tetrandra extract, and Glycyrrhiza uralensis extract being 30.8%, 4.2%, and 11.3%, respectively, and the clinical average body weight of 70 kg / person, and a dose coefficient of 12.3 between mice and humans, the mouse equivalent dose of the extract was calculated as follows: Scutellaria baicalensis extract 270, 540, and 1080 mg / day. The experiments were conducted using composition 1 (Scutellaria baicalensis extract 270 mg / kg, Stephania tetrandra extract 37.5 mg / kg, Glycyrrhiza uralensis extract 400 mg / kg) prepared in Example 1, composition 2 (Scutellaria baicalensis extract 540 mg / kg, Stephania tetrandra extract 75 mg / kg, Glycyrrhiza uralensis extract 100 mg / kg) prepared in Example 2, and composition 3 (Scutellaria baicalensis extract 1080 mg / kg, Stephania tetrandra extract 150 mg / kg, Glycyrrhiza uralensis extract 200 mg / kg) prepared in Example 3, with concentrations of 37.5, 75, and 150 mg / kg of Stephania tetrandra extract and 100, 200, and 400 mg / kg of Glycyrrhiza uralensis extract.

[0101] 2.2 Grouping and Administration Methods

[0102] After three days of acclimatization feeding, 45 Balb / c mice were randomly divided into 5 groups according to the randomization principle: normal control group, model group, composition 1 group, composition 2 group, and composition 3 group, with 9 mice in each group. The traditional Chinese medicine composition was prepared as an aqueous solution according to the dosage designed in the experimental design and administered by gavage at a dose of 0.1 mL / 10 g BW once daily for 7 consecutive days. The normal control group and the model group were given an equal volume of distilled water.

[0103] 2.3 ALI Modeling Method

[0104] On the seventh day after gavage administration, mice were immediately anesthetized with isoflurane and ALI modeled by nasal inhalation of LPS solution at a dose of 1 mg / kg. The normal control group inhaled an equal amount of physiological saline through the nasal cavity.

[0105] 2.4 Indicator Testing

[0106] Twenty-four hours after the ALI model was established using LPS, mice were anesthetized with 0.5% sodium pentobarbital, euthanized by exsanguination via the abdominal aorta, and their lungs were dissected. The total lung weight was measured, and the lung index was calculated using the formula: (lung weight / body weight) × 100. The right bronchus was ligated, and the wet weight (W) of the right lung was measured. After being placed in a 60°C oven for 24 hours, the dry weight (D) was measured, and the edema index (W / D) was calculated. The left lung was repeatedly irrigated three times with 0.8 mL of pre-cooled PBS, and the bronchoalveolar lavage fluid (BALF) was collected.

[0107] 3. Statistics and Analysis

[0108] Experimental data results are expressed as "mean ± standard deviation". This indicates that a one-way ANOVA was performed using SPSS 20.0 software, and a p-value < 0.05 was considered statistically significant.

[0109] 4. Results

[0110] The effects of the drugs on LPS-induced ALI in mice are shown in Table 3. Compared with the normal control group, the lung index, edema index, white blood cell count and protein content in bronchoalveolar lavage fluid (BALF) of the model group were significantly increased (P<0.05 or P<0.01), indicating significant lung tissue damage. Compared with the model group, all three compositions could reduce the lung index, edema index, white blood cell count and protein content in bronchoalveolar lavage fluid (BALF) to varying degrees, with composition group 2 showing the most significant effect (P<0.05 or P<0.01).

[0111] Table 3. Effects of the drug on LPS-induced ALI in mice (n=9)

[0112]

[0113]

[0114] Note: A represents Scutellaria baicalensis extract, B represents Stephania tetrandra extract, and C represents Glycyrrhiza uralensis extract, compared with the model group.

[0115] *P<0.05, **P<0.01.

[0116] 5. Conclusion

[0117] Based on the experimental results of Example 2, the preventive administration of the traditional Chinese medicine composition of the present invention can improve acute lung injury caused by LPS to varying degrees, with composition 2 showing the most significant effect.

[0118] Experiment Example 3: Therapeutic Drug Administration Experiment

[0119] 1. Experimental Materials

[0120] 1.1 Animals

[0121] Thirty-six male SPF-grade Balb / c mice, weighing 18–20 g, were purchased from Spiford (Beijing) Biotechnology Co., Ltd. and housed at the Animal Experiment Center of Xiyuan Hospital, China Academy of Chinese Medical Sciences, at a temperature of 22±2℃ and a relative humidity of 55%±2%, with 12-hour light-dark cycles. They had free access to food and water.

[0122] 1.2 Drugs and Reagents

[0123] The licorice extract, Stephania tetrandra extract, and Scutellaria baicalensis extract in the traditional Chinese medicine composition of this invention were all prepared by the Institute of Basic Research, Xiyuan Hospital, China Academy of Chinese Medical Sciences.

[0124] Lipopolysaccharide (LPS) O55: B55, a product of Sigma, USA, batch number: 0000223034.

[0125] 2. Experimental Methods

[0126] 2.1 Experimental Design Scheme

[0127] The following experiments were conducted using the herbal compositions prepared in Examples 1-7 of this invention, according to the mouse equivalent dose determined in Example 2: Composition 1 (Scutellaria baicalensis 5g, extract 270mg / kg; Stephania tetrandra 5g, extract 37.5mg / kg; Glycyrrhiza uralensis 20g, extract 400mg / kg); Composition 2 (Scutellaria baicalensis 10g, extract 540mg / kg; Stephania tetrandra 10g, extract 75mg / kg; Glycyrrhiza uralensis 5g, extract 100mg / kg); Composition 3 (Scutellaria baicalensis 20g, extract 1080mg / kg; Stephania tetrandra 20g, extract 150mg / kg; Glycyrrhiza uralensis 10g, extract 200mg / kg); Composition 4 (Scutellaria baicalensis 10g, extract 540mg / kg); Composition 5 (Scutellaria baicalensis 10g, extract 540mg / kg); Composition 6 (Scutellaria baicalensis 10g, extract 540mg / kg); Composition 7 (Stephania tetrandra 10g, extract 75mg / kg; Glycyrrhiza uralensis 10g, extract 200mg / kg); Composition 8 (Scutellaria baicalensis 10g, extract 540mg / kg); Composition 9 (Scutellaria baicalensis 10g, extract 540mg / kg); Composition 10 ... The following compositions were used in a therapeutic administration experiment to verify their effects on ALI in mice: Composition 5 (Scutellaria baicalensis 8g, extract 432mg / kg; Stephania tetrandra 8g, extract 60mg / kg; Glycyrrhiza uralensis 8g, extract 160mg / kg); Composition 6 (Scutellaria baicalensis 15g, extract 810mg / kg; Stephania tetrandra 15g, extract 112.5mg / kg; Glycyrrhiza uralensis 8g, extract 160mg / kg); Composition 7 (Scutellaria baicalensis 8g, extract 432mg / kg; Stephania tetrandra 8g, extract 60mg / kg; Glycyrrhiza uralensis 15g, extract 300mg / kg).

[0128] 2.2 Grouping and Administration Methods

[0129] Eighty-one male Balb / c mice were randomly divided into nine groups: normal control group, model group, combination group 1, combination group 2, combination group 3, combination group 4, combination group 5, combination group 6, and combination group 7, with nine mice in each group. ALI modeling was established (ALI modeling method is described in section 2.3 below; ALI modeling was not performed on the normal control group). 0.5 h after ALI modeling, the normal control group and model group were administered water by gavage at a volume of 0.1 mL / 10 g BW. The traditional Chinese medicine combination was prepared as an aqueous solution according to the dosage and administered by gavage at a volume of 0.1 mL / 10 g BW. All groups were administered the combination once daily for three consecutive days.

[0130] 2.3 ALI Modeling Method

[0131] Mice were anesthetized with isoflurane and ALI modeled by nasal inhalation of LPS solution at a dose of 1 mg / kg. The normal control group inhaled an equal amount of physiological saline through the nasal cavity.

[0132] 2.4 Indicator Testing

[0133] 72 hours after the ALI model was established using LPS, mice were anesthetized with 0.5% sodium pentobarbital, euthanized by exsanguination via the abdominal aorta, and their lungs were dissected. The total lung weight was measured, and the lung index was calculated using the formula: (lung weight / body weight) × 100. The right bronchus was ligated, and the wet weight (W) of the right lung was measured. After being placed in a 60°C oven for 24 hours, the dry weight (D) was measured, and the edema index (W / D) was calculated. The left lung was irrigated three times with 0.8 mL of pre-cooled PBS, and the bronchoalveolar lavage fluid (BALF) was collected.

[0134] The upper lobe of the right lung was fixed in 10% neutral formalin for 24 hours, then dehydrated, cleared, embedded in paraffin, and cut into 5μm thick sections. Hematoxylin-eosin (HE) staining was performed, and the lung tissue pathological lesions were observed under a light microscope, and a pathological score was calculated. The lung tissue pathological scoring criteria were as follows: 1. Degree of inflammation; 2. Alveolar wall thickness; 3. Degree of lung tissue consolidation. The severity was scored from 0 to 5 points (0: no lesion; 1: minor lesion; 2: mild lesion; 3: moderate lesion; 4: severe lesion; 5: extremely severe lesion). The scores for each item were summed to obtain the total pathological score.

[0135] 3. Statistics and Analysis

[0136] Experimental data results are expressed as "mean ± standard deviation". This indicates that a one-way ANOVA was performed using SPSS 20.0 software, and a p-value < 0.05 was considered statistically significant.

[0137] 4. Results

[0138] The pathological experimental results of LPS-induced ALI in mice are shown in the figure. Figure 1 (40X) and Figure 2(100X). The effects of the drug on LPS-induced ALI in mice are shown in Table 4. The lung index, edema index, BALF white blood cell and protein content in the model group were significantly higher than those in the normal control group (p<0.01). HE staining of lung tissue pathological sections showed that 72 h after LPS modeling, there was diffuse infiltration of neutrophils and lymphocytes in the lung tissue, extensive disappearance of alveolar structure, large-area solidification, and severe lung tissue damage. The lung index, edema index, BALF white blood cell and protein content in groups 1-7 of the composition were significantly lower than those in the model group, and the pathological score was significantly lower than that in the model group (P<0.05). This indicates that the traditional Chinese medicine composition of the present invention can significantly improve ALI and has a good protective effect against ALI.

[0139] Table 4. Effects of each drug group on LPS-induced ALI in mice (n=9)

[0140]

[0141] Note: A is Scutellaria baicalensis extract, B is Stephania tetrandra extract, and C is Glycyrrhiza uralensis extract. Compared with the model group, *P<0.05 and **P<0.01.

[0142] 5. Conclusion

[0143] Based on the experimental results of Example 3, the traditional Chinese medicine composition of the present invention can significantly improve lung tissue inflammation and edema caused by LPS and can effectively treat acute lung injury.

[0144] Embodiments of the present invention have been disclosed for illustrative purposes; however, those skilled in the art will understand that various substitutions, variations, and modifications are within the scope of the present invention without departing from the spirit and scope of the appended claims. Therefore, the scope of protection of the present invention is not limited to the contents disclosed in the embodiments.

Claims

1. A traditional Chinese medicine composition for treating acute lung injury, comprising the following ingredients: Scutellaria baicalensis, Stephania tetrandra, and Glycyrrhiza uralensis, wherein, By weight, Scutellaria baicalensis, 5-30 parts by weight; 5-30 parts by weight of Stephania tetrandra; Licorice 1-25 parts by weight; The traditional Chinese medicine composition is obtained by the following preparation method: Weigh out Scutellaria baicalensis according to the prescription ratio, and extract three times with 10-14 times the amount of water under reflux. The first extraction takes 1-3 hours, the second takes 1-2 hours, and the third takes 0.8-1.5 hours. Cool, filter, concentrate the filtrate under reduced pressure at 70-90℃, and freeze-dry to obtain crude Scutellaria baicalensis extract. Accurately weigh the above crude Scutellaria baicalensis extract, add it to macroporous adsorption resin, and elute successively with water and 50-90% ethanol. Discard the water eluent, concentrate the ethanol eluent under reduced pressure, and freeze-dry to obtain Scutellaria baicalensis extract. Weigh out the ingredients of Stephania tetrandra according to the prescription ratio, and extract three times by reflux with 8-12 times the amount of 60-90% ethanol. The first extraction takes 1-3 hours, the second takes 1-2 hours, and the third takes 1-2 hours. After cooling, filter the extract, concentrate the filtrate under reduced pressure at 70-90℃, and freeze-dry to obtain the crude extract of Stephania tetrandra. Accurately weigh the above crude extract of Stephania tetrandra, add it to a macroporous adsorption resin, and elute sequentially with water and 50-90% ethanol. Discard the water eluent, concentrate the ethanol eluent under reduced pressure, and freeze-dry to obtain the Stephania tetrandra extract. Licorice was weighed according to the prescription ratio and extracted three times by reflux with 8-12 times the amount of 60-90% ethanol. The first extraction was for 1-3 hours, the second for 1-2 hours, and the third for 1-2 hours. After cooling, the extract was filtered, and the filtrate was concentrated under reduced pressure at 70-90℃ and freeze-dried to obtain crude licorice extract. The crude licorice extract was accurately weighed and added to a macroporous adsorption resin. The resin was eluted sequentially with water and 50-90% ethanol. The water eluent was discarded, and the ethanol eluent was concentrated under reduced pressure and freeze-dried to obtain licorice extract. The above-mentioned Scutellaria baicalensis extract, Stephania tetrandra extract and Glycyrrhiza uralensis extract are mixed to obtain the traditional Chinese medicine composition.

2. The traditional Chinese medicine composition according to claim 1, wherein, By weight, Scutellaria baicalensis, 5-20 parts by weight; 5-20 parts by weight of Stephania tetrandra; Licorice 5-20 parts by weight.

3. A method for preparing the traditional Chinese medicine composition as described in claim 1 or 2, comprising the following steps: Weigh out Scutellaria baicalensis according to the prescription ratio, and extract three times with 10-14 times the amount of water under reflux. The first extraction takes 1-3 hours, the second takes 1-2 hours, and the third takes 0.8-1.5 hours. Cool, filter, concentrate the filtrate under reduced pressure at 70-90℃, and freeze-dry to obtain crude Scutellaria baicalensis extract. Accurately weigh the above crude Scutellaria baicalensis extract, add it to macroporous adsorption resin, and elute successively with water and 50-90% ethanol. Discard the water eluent, concentrate the ethanol eluent under reduced pressure, and freeze-dry to obtain Scutellaria baicalensis extract. Weigh out the ingredients of Stephania tetrandra according to the prescription ratio, and extract three times by reflux with 8-12 times the amount of 60-90% ethanol. The first extraction takes 1-3 hours, the second takes 1-2 hours, and the third takes 1-2 hours. After cooling, filter the extract, concentrate the filtrate under reduced pressure at 70-90℃, and freeze-dry to obtain the crude extract of Stephania tetrandra. Accurately weigh the above crude extract of Stephania tetrandra, add it to a macroporous adsorption resin, and elute sequentially with water and 50-90% ethanol. Discard the water eluent, concentrate the ethanol eluent under reduced pressure, and freeze-dry to obtain the Stephania tetrandra extract. Licorice was weighed according to the prescription ratio and extracted three times by reflux with 8-12 times the amount of 60-90% ethanol. The first extraction was for 1-3 hours, the second for 1-2 hours, and the third for 1-2 hours. After cooling, the extract was filtered, and the filtrate was concentrated under reduced pressure at 70-90℃ and freeze-dried to obtain crude licorice extract. The crude licorice extract was accurately weighed and added to a macroporous adsorption resin. The resin was eluted sequentially with water and 50-90% ethanol. The water eluent was discarded, and the ethanol eluent was concentrated under reduced pressure and freeze-dried to obtain licorice extract. The above-mentioned Scutellaria baicalensis extract, Stephania tetrandra extract and Glycyrrhiza uralensis extract are mixed to obtain the traditional Chinese medicine composition.

4. The preparation method according to claim 3, wherein, In the preparation steps of Scutellaria baicalensis extract, the macroporous adsorption resin used is selected from one or more of D101, HPD-300, AB-8, HPD-600 and HPD-826; the mass ratio of Scutellaria baicalensis to macroporous adsorption resin is 1:2 to 1:

4.

5. The preparation method according to claim 3 or 4, wherein, In the preparation steps of Stephania tetrandra extract, the macroporous adsorption resin used is selected from one or more of D101, HPD-300, AB-8, HPD-600 and HPD-826; the mass ratio of Stephania tetrandra to macroporous adsorption resin is 1:0.5 to 1:0.

8.

6. The preparation method according to claim 3 or 4, wherein, In the preparation of licorice extract, the macroporous adsorption resin used is selected from one or more of D101, HPD-300, AB-8, HPD-600 and HPD-826; the mass ratio of licorice to macroporous adsorption resin is 1:1 to 1:

2.

7. Use of a traditional Chinese medicine composition as described in claim 1 or 2 in the preparation of a medicament for treating and / or preventing acute lung injury.

8. A pharmaceutical preparation comprising a traditional Chinese medicine composition as described in claim 1 or 2 and a pharmaceutically acceptable carrier.

9. The pharmaceutical preparation according to claim 8, wherein, The pharmaceutical preparation is administered orally, intravenously, locally, by inhalation or spraying, sublingually, transdermally, or rectally.

10. The pharmaceutical preparation according to claim 8, wherein, The pharmaceutical preparation is selected from tablets, granules, hard capsules, mixtures, pills, injections, or soft capsules.

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