Composition for treating idiopathic pulmonary fibrosis and preparation method and application thereof

Abstract

The invention discloses a composition for treating idiopathic pulmonary fibrosis and a preparation method and application thereof. The composition for treating idiopathic pulmonary fibrosis is formed by mixing baicalin nanocrystals and ambroxol hydrochloride, and is prepared into various medical dosage forms through a conventional method, including oral preparations such as tablets, capsules, granules or oral liquid. The composition can remarkably improve rat idiopathic pulmonary fibrosis caused by bleomycin sulfate, relieve the symptom of rat pulmonary fibrosis by reducing the inflammatory factor content, the total protein content and collagen deposition, and improve the oral bioavailability of the monomer baicalin. The traditional Chinese medicine composition is suitable for preventing and treating idiopathic pulmonary fibrosis.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a composition for treating idiopathic pulmonary fibrosis, a preparation method thereof and use thereof. Background Art

[0002] Idiopathic pulmonary fibrosis is a chronic and irreversible lung disease with diffuse alveolitis and alveolar structural disorder, which is prone to occur in middle-aged and elderly people. It is characterized by extensive lung remodeling caused by abnormal deposition of extracellular matrix, and eventually leads to pulmonary fibrosis. The development of idiopathic pulmonary fibrosis includes lung damage, inflammation, the formation of myofibroblasts and the accumulation of extracellular matrix, which eventually lead to lung structural dysfunction. The clinical symptoms are insidious onset, and the early symptoms are not obvious. Initially, they present as cough and sputum, and later, dyspnea worsens, leading to respiratory failure and death. It has been found in clinical practice that idiopathic pulmonary fibrosis is difficult to treat, and the drugs used are expensive. They are only suitable for short-term or acute use, have large toxic side effects, and are not suitable for long-term administration. Therefore, the development of a drug that can prevent / treat idiopathic pulmonary fibrosis has important social significance. Summary of the invention

[0003] In view of the many limitations of current drugs on the market, the present invention aims to provide a composition for treating idiopathic pulmonary fibrosis to improve the situation of patients with idiopathic pulmonary fibrosis. The drug of the present invention uses baicalin nanocrystals and ambroxol hydrochloride as main raw materials, has a simple preparation process, significant efficacy, high safety, and is suitable for the treatment of idiopathic pulmonary fibrosis.

[0004] To achieve the above object, the present invention provides the following technical solutions: A Chinese medicine composition for treating idiopathic pulmonary fibrosis, comprising baicalin nanocrystals and ambroxol hydrochloride (the main component baicalin has a structural formula as shown in Figure 1 As shown in A, the molecular formula of ambroxol hydrochloride is as follows Figure 1 (shown in B).

[0005] The baicalin nanocrystals are nanoparticles formed by baicalin and poloxamer 188, the baicalin content in the baicalin nanocrystals is 65-85%, preferably 85%, and the particle size of the baicalin nanocrystals is 330-400nm.

[0006] Preferably, baicalin nanocrystals are prepared according to the following method: The baicalin raw material is weighed, placed in an aqueous solution containing 0.1%-0.2% poloxamer P188, and placed in a high-pressure homogenizer after ultrasonic pre-dispersion. After 5 full cycles at 300 bar, 25 cycles at 1000 bar are performed to obtain a nanocrystalline suspension, which is freeze-dried to obtain baicalin nanocrystals. Preferably, the content of baicalin in the obtained baicalin nanocrystals is 65-85%, preferably 85%; preferably, the weight ratio of baicalin nanocrystals to ambroxol hydrochloride in the composition is 1:0.1-0.3, preferably 1:0.2.

[0007] Furthermore, the present invention also proposes the use of the composition for treating idiopathic pulmonary fibrosis in the preparation of a drug for treating idiopathic pulmonary fibrosis; wherein, preferably, the composition can be prepared into various oral preparations according to conventional preparation methods; Among them, preferably, the oral preparations include tablets, granules, capsules, and oral liquids.

[0008] Experiments have shown that the composition for treating idiopathic pulmonary fibrosis of the present invention can significantly improve idiopathic pulmonary fibrosis in rats caused by bleomycin sulfate, and at the same time reduce the content of inflammatory factors, total protein content and collagen deposition to alleviate the symptoms of pulmonary fibrosis in rats, and has potential development value for the prevention and treatment of idiopathic pulmonary fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 The structural formulas of baicalin and ambroxol hydrochloride, where A is baicalin; B is ambroxol hydrochloride.

[0010] Figure 2 HE and MASSON staining results (200X), where AF are the HE staining results of rat lung tissue: A blank group; B model group; C positive drug group; D baicalin nanocrystal group; E ambroxol hydrochloride group; F combination group; GL are the MASSON staining results of rat lung tissue: G blank group; H model group; I positive drug group; J baicalin nanocrystal group; K ambroxol hydrochloride group; L combination group.

[0011] Figure 3 Body weight changes of rats over 28 days. Figure 4 MICRO-CT images of rats.

[0012] Among them, A is the blank group; B is the model group; C is the positive drug group; D is the baicalin nanocrystal group; E is the ambroxol hydrochloride group; and F is the combination group.

[0013] Figure 5 Rat lung coefficient levels.

[0014] Figure 6Total protein levels in rat lung homogenates.

[0015] Figure 7 Levels of IL-6, IL-1β, IL-4, and TGF-β1 in rat lung tissue homogenates.

[0016] Figure 8 Lung function levels in rats. DETAILED DESCRIPTION

[0017] Example 1 1) Weigh the baicalin raw material, place it in an aqueous solution containing 0.1% poloxamer 188, pre-disperse it by ultrasonication, place it in a high-pressure homogenizer, cycle it at 300 bar for 5 times, and then cycle it at 1000 bar for 25 times to obtain a nanocrystal suspension, and freeze-dry it to obtain baicalin nanocrystals; 2) According to the weight ratio of baicalin nanocrystals to ambroxol hydrochloride being 1:0.2, weigh baicalin nanocrystals and ambroxol hydrochloride, and mix them to obtain a baicalin nanocrystal.

[0018] Example 2 1) Weigh the baicalin raw material, place it in an aqueous solution containing 0.15% poloxamer 188, pre-disperse it by ultrasonication, place it in a high-pressure homogenizer, cycle it at 300 bar for 5 times, and then cycle it at 1000 bar for 25 times to obtain a nanocrystal suspension, and freeze-dry it to obtain baicalin nanocrystals; 2) According to the weight ratio of baicalin nanocrystals to ambroxol hydrochloride being 1:0.2, weigh baicalin nanocrystals and ambroxol hydrochloride, and mix them to obtain a baicalin nanocrystal.

[0019] Example 3 1) Weigh the baicalin raw material, place it in an aqueous solution containing 0.2% poloxamer 188, pre-disperse it by ultrasonication, place it in a high-pressure homogenizer, cycle it at 300 bar for 5 times, and then cycle it at 1000 bar for 25 times to obtain a nanocrystal suspension, and freeze-dry it to obtain baicalin nanocrystals; 2) According to the weight ratio of baicalin nanocrystals to ambroxol hydrochloride being 1:0.2, weigh baicalin nanocrystals and ambroxol hydrochloride, and mix them to obtain a baicalin nanocrystal.

[0020] Example 4 1) Weigh the baicalin raw material, place it in an aqueous solution containing 0.15% poloxamer 188, pre-disperse it by ultrasonication, place it in a high-pressure homogenizer, cycle it at 300 bar for 5 times, and then cycle it at 1000 bar for 25 times to obtain a nanocrystal suspension, and freeze-dry it to obtain baicalin nanocrystals; 2) According to the weight ratio of baicalin nanocrystals to ambroxol hydrochloride being 1:0.1, weigh baicalin nanocrystals and ambroxol hydrochloride, and mix them to obtain a baicalin nanocrystal.

[0021] Example 5 1) Weigh the baicalin raw material, place it in an aqueous solution containing 0.15% poloxamer 188, pre-disperse it by ultrasonication, place it in a high-pressure homogenizer, cycle it at 300 bar for 5 times, and then cycle it at 1000 bar for 25 times to obtain a nanocrystal suspension, and freeze-dry it to obtain baicalin nanocrystals; 2) According to the weight ratio of baicalin nanocrystals to ambroxol hydrochloride being 1:0.3, weigh baicalin nanocrystals and ambroxol hydrochloride, and mix them to obtain a baicalin nanocrystal.

[0022] Experimental Example 1 Baicalin Nanocrystals 1.1 Instruments and reagents High-pressure homogenizer (AH00D, ATS Industrial Systems Co., Ltd.); Malvern particle size and potential analyzer (Nano-ZS, Marlvern); ultrasonic cleaning machine (KQ-400KDE, Kunshan Ultrasonic Instrument Co., Ltd.); high-performance liquid chromatograph (Agilent-1260, Agilent Technologies Co., Ltd.); one-millionth balance (XPR6UD50, Mettler-Toledo); one-tenth balance (AX224ZH, Ohaus Instrument (Changzhou) Co., Ltd.).

[0023] Baicalin (JS252432, ≥90%, Shanghai Yuanye Biotechnology Co., Ltd.); Ambroxol hydrochloride (S44913, ≥98%, Shanghai Yuanye Biotechnology Co., Ltd.); Baicalin reference substance (N15GB167969, ≥98%, Shanghai Yuanye Biotechnology Co., Ltd.); Poloxamer (BASF Co., Ltd.); Acetonitrile (chromatographic grade, Tianjin Concord Reagent Co., Ltd.); Ethanol (analytical grade, Tianjin Concord Reagent Co., Ltd.); Phosphoric acid (218128, Thermo Fisher Scientific (China) Co., Ltd.).

[0024] 1.2 Preparation method The baicalin raw material was weighed and placed in an aqueous solution containing 0.15% poloxamer 188. After ultrasonic pre-dispersion, it was placed in a high-pressure homogenizer. After 5 full cycles at 300 bar and 25 cycles at 1000 bar, a nanocrystalline suspension was obtained with a measured particle size of 350 nm. Baicalin nanocrystals were obtained by freeze-drying. After re-dissolution, the particle size was measured to be 370 nm, with no obvious change.

[0025] 1.3 Content determination Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; 0.1% phosphoric acid water (A)-acetonitrile (B) solution was used as mobile phase; column temperature was 20°C; detection wavelength was 237nm (baicalin). The elution gradient was 0-3min, 15%B; 3-8min, 15%-25%B; 8-11min, 25%-30%B; 11-15min, 30%B.

[0026] Preparation of reference solution: Accurately measure 5.50 mg of baicalin reference substance, place it in a 5 mL volumetric flask and dissolve it in methanol to obtain a mixed reference solution with a concentration of 1.10 mg / mL.

[0027] Preparation of the test solution: Take 5 mg of the nanocrystals obtained by freeze-drying, accurately add 25 ml of methanol, ultrasonicate for 10 min, and take the filtrate to obtain the test solution.

[0028] It was determined that the content of baicalin in the prepared baicalin nanocrystals was 84.54%.

[0029] Experimental Example 2 Baicalin Nanocrystal-Ambroxol Hydrochloride Composition According to the weight ratio of baicalin nanocrystals to ambroxol hydrochloride being 1:0.2, baicalin nanocrystals (prepared according to Experimental Example 1) and ambroxol hydrochloride were weighed and mixed to obtain the product.

[0030] 4 Experimental part: Pharmacodynamic study of the composition prepared in Experimental Example 2 on idiopathic pulmonary fibrosis in rats 4.1 Experimental Animals and Test Drugs 4.1.1 Drug testing Baicalin nanocrystals (prepared according to Experimental Example 1); composition (prepared according to Experimental Example 2); physiological saline (G4702, Wuhan Servicebio Company); 4% tissue fixative (P1110, Beijing Solebow Technology Co., Ltd.).

[0031] Pirfenidone capsules (National Medicine Standard No. H20133376, Beijing Contini Pharmaceutical Co., Ltd.); Bleomycin sulfate (B107423-100mg, Shanghai Aladdin Biochemical Technology Co., Ltd.); Rat BCA protein quantitative detection kit (060723240430, Shanghai Biyuntian Biotechnology Co., Ltd.); Rat interleukin-6 (IL-6), rat interleukin-1β (IL-1β), rat interleukin-4 (IL-4), rat transforming growth factor (TGF-β1) ELISA kits (MM-0190R1, MM-0047R1, MM-0191R1, MM-0181R1, Jiangsu Enzyme Immunity Industry Co., Ltd.); Chloraldehyde hydrate (302-17-0, Shanghai Aladdin Biochemical Technology Co., Ltd.).

[0032] 4.1.2 Experimental animals 60 male SD rats weighing 180-220 g 4.2 Experimental methods 4.2.1 Establishment of rat pulmonary fibrosis model The rat pulmonary fibrosis model was established by intratracheal instillation of bleomycin sulfate. After anesthetizing the rats, they were fixed on a board tilted at 35° in a supine position, and the neck skin was routinely disinfected with 75% medical alcohol. Then, a 2 to 3 cm skin incision was made in the midline of the neck, and the muscles were bluntly separated using vascular clamps to expose the trachea. Next, a No. 4 needle was used to penetrate the gap between the tracheal cartilage rings, puncture the trachea toward the heart, and inject about 0.3 mL of a solution containing bleomycin sulfate (at a dose of 5 mg / kg) through an empty syringe. After the injection, the mouse board was quickly rotated vertically and placed upright for 3 min to allow the drug to be evenly distributed in the lungs. After that, the muscles and skin were sutured and disinfected again. The blank control group rats only received an intratracheal injection of an equal amount of normal saline, and other operations were the same as those of the modeling group. Except for the blank control group, all other groups were made into pulmonary fibrosis models.

[0033] 4.2.2 Animal grouping and treatment After 3 days of adaptive feeding, 60 SD rats were randomly divided into 6 groups: blank group (Blank), model group (Model), pirfenidone capsule positive drug group (PFD, 52 mg / kg), baicalin nanocrystal group (BA, 60 mg / kg), ambroxol hydrochloride group (AH, 18 mg / kg) and baicalin nanocrystal-ambroxol hydrochloride combination group (BA-AH, 78 mg / kg). Oral gavage was started on the second day of modeling and continued once a day until the end of 28 days.

[0034] 4.2.3 Sample collection and storage The rats' breathing, activity, eating and body weight were observed daily. The bleomycin sulfate model was regarded as the first day. The body weights of the rats in each group were recorded on days 1, 7, 14, 21 and 28, and statistical analysis was performed. MICRO-CT imaging examination of rats was performed on day 28 of modeling, and various respiratory indicators of rats in free-moving state were tested with EMKA pulmonary function tester, including duration of inspiration (TI), duration of expiration (TE), respiratory rate (f), maximum inspiratory flow (PIF), maximum expiratory flow (PEF), tidal volume (TV), relaxation time (RT), bronchial contraction parameter (Penh), etc. After the test, blood was collected from the abdominal aorta and the rats were killed. The alveolar lavage fluid of the rats was collected, and the lung tissue was removed and weighed. The left lung was immersed in 10% neutral formalin fixative, and HE staining and MASSON staining were performed 24 hours later to observe the lesions of lung tissue.

[0035] 4.2.4 Rat lung imaging examination The model of in vivo Micro CT scanning was Quantum FX, and the scanning parameters were as follows: tube voltage: 90kV, tube current: 88uA, imaging field of view: 72*40 mm, pixel size of reconstructed image: 144 um. The in vivo rats were placed in prone position with their limbs spread out and head forward, anesthetized with isoflurane, and whole lung scans were performed under free breathing to obtain micro CT images of the rat lungs.

[0036] 4.2.5 Rat lung coefficient determination Take the lung tissue sample in "4.2.3" and calculate the lung coefficient using the following formula and perform statistical analysis: Lung coefficient = lung wet weight (g) / body weight (kg) * 100% 4.2.6 Determination of total protein in rat lung tissue homogenate Take the lung tissue homogenate sample in "4.2.3" and centrifuge it at 4°C and 4000 rpm for 10 min to obtain the supernatant. Determine the total protein content by colorimetry according to the instructions of the total protein determination kit.

[0037] 4.2.7 Determination of IL-6, IL-1β, IL-4 and TGF-β1 contents in rat lung tissue homogenates Take the lung tissue homogenate sample in "4.2.3" and centrifuge it at 4°C and 4000 rpm for 10 min to obtain the supernatant. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of inflammatory factors IL-4, IL-6, TNF-β1, and IL-1β. The specific experimental operation was carried out according to the steps in the instructions of the relevant ELISA kit.

[0038] 4.2.8 Statistics and Plotting The experimental data were analyzed by one-way ANOVA using SPSS 27 software and plotted using Graphpad Prism 8.0.

[0039] 4.3 Experimental Results 4.3.1 Observation of rat physiological status The rats in the blank group had a good mental state, stable breathing, regular diet, shiny fur, and no coughing or nasal secretions. The rats in the model group had rapid breathing, poor mental state, severe hair loss, coughing and nasal secretions, and reduced food intake. The different drug-administered groups showed different degrees of improvement compared with the model group. As the drug administration time increased, the rats' breathing gradually stabilized, nasal secretions decreased, and their diet, hair loss, and mental state recovered to varying degrees. The weight changes of the rats in each group over the days are as follows: Figure 3As shown, the body weight of the model group decreased significantly compared with the blank group on the 7th day (***p<0.001); on the 14th day, the body weight of the rats in the positive drug group, baicalin nanocrystal group and combination group 1 began to increase significantly compared with the model group (n=10, #p<0.05, ##p<0.01, ###p<0.001), and the trend was maintained until the end of the 28th day; from the 14th day, the body weight of the combination group increased significantly compared with the ambroxol hydrochloride group (n=10, △p<0.05), and the trend was maintained until the end of the 28th day; on the 28th day, the combination group showed a significant difference compared with the baicalin nanocrystal group (n=10, ▲p<0.05). The results show that the combination of baicalin nanocrystals and ambroxol hydrochloride has more advantages than the separate administration of baicalin nanocrystals and ambroxol hydrochloride, and can significantly improve the weight loss of rats caused by IPF.

[0040] 4.3.2 Staining of rat lung tissue sections HE-stained and MASSON-stained sections of rat lung tissue were obtained and observed under an optical microscope.

[0041] HE staining results showed that the blank group ( Figure 2 A) The lung tissue capsule structure is clear, and the lung substance is the numerous alveoli at the ends of the bronchial branches at all levels in the lungs, without collagen fiber hyperplasia; model group ( Figure 2 B) in the figure shows flake-like alveolar consolidation, unclear alveolar structure, more lymphocytes, granulocytes and macrophages (green arrows) in the alveolar wall and alveolar cavity, and occasional necrotic cell fragments (black arrows); occasional alveolar dilatation (dark blue arrows); occasional focal infiltration of lymphocytes around blood vessels and bronchioles (blue arrows); occasional perivascular edema (purple arrows), loosely arranged connective tissue, widened spaces, with a small amount of punctate infiltration of lymphocytes; occasional hydropic degeneration of bronchiolar epithelial cells (red arrows), swollen cells, and loose and lightly stained cytoplasm; positive drug group ( Figure 2 C) The lung tissue capsule structure is clear, the lung substance is the numerous alveoli at the ends of the bronchial branches at all levels in the lungs, the alveolar wall is composed of a single layer of epithelium, focal alveolar wall capillary congestion (orange arrow), rare vascular congestion (orange arrow), a small amount of brown-yellow pigment deposition in the alveolar wall (brown arrow), showing mild lesions; Baicalin nanocrystal group ( Figure 2 D) A small number of foam cells and mast cells in the alveoli (dark red arrows), occasional perivascular edema (purple arrows), loosely arranged connective tissue, widened spaces, with a small amount of lymphocyte punctate infiltration, and a small amount of eosinophilic tissue fluid (gray arrows), more vascular congestion (orange arrows), occasional brown-yellow pigment deposition (brown arrows), showing mild lesions; ambroxol hydrochloride group ( Figure 2E) in the alveoli, a small amount of foam cells (dark red arrows); a small amount of eosinophilic tissue fluid in the alveoli (gray arrows); a small amount of alveolar dilation (dark blue arrows); occasional perivascular edema (purple arrows), loose arrangement of connective tissue, widened spaces, accompanied by a small amount of lymphocyte infiltration in dots (blue arrows); irregular arrangement of bronchiolar epithelial cells (light green arrows), a small amount of epithelial cells and eosinophilic substances can be seen in the lumen, showing moderate lesions; the composition group ( Figure 2 F) in the alveolar wall, a small amount of granulocyte infiltration (green arrows); multifocal mild thickening of the alveolar wall, widened alveolar septum; a small amount of foam cells in the alveoli (dark red arrows); occasional alveolar dilation (dark blue arrows); occasional deposition of brownish-yellow pigment (brown arrows); rare vascular congestion (orange arrows), showing mild lesions.

[0042] The results of MASSON staining showed that in the blank group ( Figure 2 G) no obvious collagen fiber hyperplasia was seen in the lung tissue; in the model group ( Figure 2 H) in the lung tissue, multi-focal hyperplasia of collagen fibers (yellow arrows) was seen, and small areas of fibrous masses were formed in many places, with uneven thickness of the fibers and disordered arrangement; in the positive drug group ( Figure 2 I) in the lung tissue, a small amount of local collagen fiber hyperplasia (yellow arrows) was seen, the collagen fibers were thin, in discontinuous cord-like shapes, and arranged irregularly; in the baicalin nanocrystal group ( Figure 2 J) in the lung tissue, multi-focal hyperplasia of collagen fibers (yellow arrows) was seen, the collagen fibers were thin, in discontinuous cord-like shapes, and arranged irregularly; in the ambroxol hydrochloride group ( Figure 2 K) in the lung tissue, a relatively large range of collagen fiber hyperplasia (yellow arrows) was seen, and small areas of fibrous masses were formed in many places, with uneven thickness of the fibers and disordered arrangement; in the composition group ( Figure 2 L) in the lung tissue, a small amount of local collagen fiber hyperplasia (yellow arrows) was seen, the collagen fibers were thin, in discontinuous cord-like shapes, and arranged irregularly.

[0043] Degree of lesion: model group > ambroxol hydrochloride group > baicalin nanocrystal group > composition group ≥ positive drug group.

[0044] 4.3.3 MICRO-CT images of rat lungs As Figure 4As shown, the density of bilateral lung parenchyma in the blank group is uniform, the lung texture is clear and evenly distributed, and the bronchial bundle and vascular bundle can be seen from the inside to the outside of the lung, gradually becoming thinner, and the texture shadow formed by the regular running. No abnormal high-density shadows were found in the bilateral lung field of vision; the lung parenchyma density in the model group increased, the texture in the lung was blurred, and large nodular shadows or mass-like shadows were seen, the structure in the lung was disordered, and the lung interstitial structure was abnormal; the other drug-treated groups all showed different degrees of improvement compared with the model group. It can be seen from the figure that the severity of fibrosis is in the following order: model group > ambroxol hydrochloride group > baicalin nanocrystal group > positive drug group > combination group.

[0045] 4.3.4 Rat lung coefficient levels like Figure 5 As shown, the lung coefficients of rats in different groups were measured, and the expression of pulmonary edema in the lung tissues of rats in different groups was observed. As shown in the figure, compared with the blank group, the lung coefficient of the model group increased significantly (n=10, ***p<0.001), and the positive drug group, baicalin nanocrystal group, and combination group decreased significantly compared with the model group (n=10, #p<0.05, ##p<0.01); compared with the ambroxol hydrochloride group, the lung coefficient of the combination group decreased significantly (n=10, △p<0.05).

[0046] 4.3.5 Total protein levels in rat lung tissue homogenates like Figure 6 As shown in the figure below, the total protein content in the bronchoalveolar lavage fluid of rats in each group was determined by the BCA method. As shown in the figure below, the total protein content of the blank group was (7.99±1.50) mg / mL, and the total protein content of the model group was (17.11±1.34) mg / mL. The model group was significantly increased compared with the blank group (n=10, ***p<0.001). Compared with the model group, the total protein content of rats in the positive drug group, baicalin nanocrystal group and combination group was significantly reduced (n=10, #p<0.05, ##p<0.01). Compared with the baicalin nanocrystal group and ambroxol hydrochloride group, the total protein content of the combination group was significantly reduced (n=10, ▲p<0.05, △p<0.05).

[0047] 4.3.6 Determination of IL-4, IL-6, IL-1β and TGF-β1 contents in rat lung tissue homogenate like Figure 7As shown, the release and expression of IL-4, IL-6, IL-1β and TGF-β1 in the lung tissue homogenate of rats in different groups were measured. As shown in the figure, the contents of IL-4, IL-6, IL-1β and TGF-β1 in the model group were significantly increased compared with the blank group (n=10, ***p<0.001); the contents of IL-4, IL-6, IL-1β and TGF-β1 in the positive drug group, baicalin nanocrystal group and combination group were significantly reduced compared with the model group (n=10, #p<0.05, ##p<0.01, ###p<0.001). Compared with the baicalin nanocrystal group and the ambroxol hydrochloride group, the contents of IL-6, IL-1β and TGF-β1 in the combination group were significantly reduced (n=10, ▲p<0.05, △p<0.05, △△p<0.01).

[0048] 4.3.7 Rat lung function level like Figure 8 As shown in the lung function indexes, compared with the blank group, the lung function parameters of the model group rats Ti (inspiratory duration), Te (expiratory duration), PIF (maximum inspiratory flow), TV (tidal volume), MV (minute volume), RT (relaxation time) were significantly reduced, Penh (bronchial contraction parameter) and f (respiratory frequency) were significantly increased (n=10, ***p<0.001); compared with the model group, the respiratory indexes of the drug-treated group showed significant improvement (n=10, #p<0.05, ##p<0.01, ###p<0.001), except for TV and f, in the remaining lung function indexes. Compared with the baicalin nanocrystal group and the ambroxol hydrochloride group, the combination group showed significant differences (n=10, ▲p<0.05, ▲▲p<0.01, △p<0.05, △△p<0.01).

[0049] By measuring the content of total protein and inflammatory factors, it can be seen that the combination group can alleviate the symptoms of pulmonary fibrosis in rats by reducing the content of inflammatory factors, total protein content and collagen deposition, and also has a certain therapeutic effect on the treatment of pulmonary fibrosis; from the perspective of body weight, total protein, inflammatory factors and lung function, it can be seen that compared with the baicalin nanocrystal single-drug group and the ambroxol hydrochloride group, the combination effect of the combination group is more significant, and there is no significant difference compared with the positive drug group, but from the perspective of inflammatory factors, total protein content and lung function indicators, it can be seen that the combination group is slightly lower than the positive drug group, proving that it has a decreasing trend.

Claims

1. A composition for treating idiopathic pulmonary fibrosis, characterized in that: It is composed of baicalin nanocrystals and ambroxol hydrochloride.

2. The composition according to claim 1, characterized in that The baicalin nanocrystals are nanoparticles prepared by high-pressure homogenization method by placing baicalin in a poloxamer 188 aqueous solution with a mass percentage concentration of 0.1%-0.2%.

3. The composition according to claim 2, characterized in that The mass percentage concentration of the poloxamer 188 aqueous solution is 0.1%.

4. The composition according to claim 2, characterized in that The mass percentage concentration of the poloxamer 188 aqueous solution is 0.15%.

5. The composition according to claim 2, characterized in that The mass percentage concentration of the poloxamer 188 aqueous solution is 0.2%.

6. The composition according to any one of claims 1 to 5, characterized in that The preparation method of baicalin nanocrystals comprises the following steps: (1) Weigh baicalin, place it in an aqueous solution of poloxamer 188, and pre-disperse it by ultrasonication; (2) adding the pre-dispersed mixture into a high-pressure homogenizer, circulating it for 5 times at 300 bar, and then circulating it for 25 times at 1000 bar to obtain a nanocrystal suspension; (3) The nanocrystal suspension is freeze-dried to obtain baicalin nanocrystals.

7. The composition according to any one of claims 1 to 5, characterized in that The content of baicalin in the baicalin nanocrystals is 65-85%.

8. The composition according to any one of claims 1 to 5, characterized in that The obtained baicalin nanocrystals have a particle size of 330-400 nm.

9. The composition according to any one of claims 1 to 5, characterized in that: The composition is prepared by mixing baicalin nanocrystals and ambroxol hydrochloride in a mass ratio of 1:0.1-0.

3.

10. The composition according to claim 9, characterized in that: The mass ratio of baicalin nanocrystals to ambroxol hydrochloride is 1:0.

1.

11. The composition according to claim 9, characterized in that: The mass ratio of baicalin nanocrystals to ambroxol hydrochloride is 1:0.

2.

12. The composition according to claim 9, characterized in that: The mass ratio of baicalin nanocrystals to ambroxol hydrochloride is 1:0.

3.

13. Use of the composition according to any one of claims 1 to 12 in the preparation of a medicament for treating idiopathic pulmonary fibrosis.

14. The use according to claim 13, characterized in that: The composition is prepared into an oral preparation.

15. The use according to claim 14, characterized in that: Oral preparations include tablets, granules, capsules, and oral solutions.

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