Use of a triacetyl andrographolide and its composition in the preparation of a drug for relaxing airway smooth muscle

Abstract

The application discloses application of triacetylandrographolide and a composition thereof in preparation of a medicine for relaxing airway smooth muscle, and belongs to the technical field of medicines. In view of the problems of obvious side effects and narrow treatment window of existing medicines, the application finds that triacetylandrographolide (3, 14, 19-Triacetylandrographolide, TAA) and the composition thereof have tracheal relaxation activity, and the triacetylandrographolide and the composition thereof play a relaxation role through non-competitive antagonism of a downstream signal path of a cholinergic receptor. The new mechanism provides a new idea for developing a new bronchial dilator for overcoming drug resistance of existing medicines. The TAA after preparation optimization can be used for treating or preventing airway obstructive diseases such as asthma, chronic obstructive pulmonary disease, bronchitis and acute lung injury.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and specifically relates to the application of triacetylandrographolide. Background Technology

[0002] Asthma and chronic obstructive pulmonary disease (COPD) are common chronic respiratory diseases, characterized by excessive contraction of tracheal smooth muscle, leading to airway narrowing and airflow restriction. Currently, commonly used bronchodilators include β2-adrenergic receptor agonists (such as salbutamol), anticholinergic drugs (such as ipratropium bromide), and theophylline. While these drugs effectively relieve symptoms, they also have drawbacks such as sinus tachycardia, muscle tremors, dry mouth, constipation, and a narrow therapeutic window. Therefore, the development of novel bronchodilators with novel chemical structures and low side effects has become an urgent need in this field.

[0003] Natural products, due to their structural diversity and rich bioactivity, have become an important source for the discovery of drug lead compounds. Triacetylandrographolide (TAA) is a diterpenoid lactone natural framework compound extracted from the traditional Chinese medicine Andrographis paniculata. Existing patents, including CN1695612A, disclose that TAA possesses anti-inflammatory, immunosuppressive, and antitumor activities; and CN118236329A discloses a TAA nanocrystal, its preparation method, and its applications, demonstrating that it can significantly improve pulmonary ventilation, reduce the number of inflammatory cells and the secretion of inflammatory factors in the lungs, and inhibit the infiltration of inflammatory cells in lung tissue, making it suitable for the treatment of lung injury-related diseases. However, whether TAA has a direct bronchodilatory effect on tracheal smooth muscle has not been reported in domestic or international literature or patent databases. Summary of the Invention

[0004] This invention addresses the technical problems of existing bronchodilators having significant side effects or a narrow safety window by proposing the application of triacetylandrographolide and its composition in the preparation of drugs that relax airway smooth muscle.

[0005] To achieve the above objectives, the technical solution of the present invention is implemented as follows:

[0006] In a first aspect, the present invention provides the use of 3,14,19-Triacetylandrographolide (TAA) in the preparation of drugs that relax airway smooth muscle.

[0007] This invention provides the use of triacetylandrographolide in the preparation of drugs for the treatment or prevention of asthma.

[0008] This invention provides the use of triacetylandrographolide in the preparation of medicaments for treating chronic obstructive pulmonary disease.

[0009] The triacetylandrographolide exerts its vasodilatory effect by non-competitively antagonizing downstream signaling pathways of cholinergic receptors.

[0010] The present invention has confirmed the above-mentioned uses through in vivo and in vitro pharmacodynamic experiments.

[0011] In vitro rat tracheal ring experiments showed that TAA, within a concentration range of 30-100 μM, could relax tracheal smooth muscle pre-contracted by carbacholine in a concentration-dependent manner, with a maximum relaxation rate of 100.8 ± 7.3%, and had no significant effect on the resting tracheal basal tension. Preliminary mechanistic studies showed that TAA (30 μM) could significantly reduce the carbacholine-induced maximum contractile effect (E0.05). max (From 222.6 ± 9.9% to 141.1 ± 10.1%) and improved its EC 50 (Increased from 0.15 μM to 0.70 μM), suggesting that it may exert its effects through non-competitive antagonism of downstream cholinergic receptor pathways.

[0012] TAA nanocrystals also exhibited a concentration-dependent vasodilatory effect in the concentration range of 1–100 μM, with a maximum vasodilation rate of 76.5 ± 6.5%, and their effect was significantly better than that of solvent and excipients at 30–100 μM (p < 0.01). They can significantly reduce the E6 of carbacholine. max (From 222.6 ± 9.9% to 151.2 ± 13.2%) and improved EC 50 (Increased from 0.15 μM to 0.38 μM).

[0013] In a rat airway hyperresponsiveness model, administration of TAA nanocrystals (20 mg / kg) via the airway significantly inhibited acetylcholine-induced airway hyperresponsiveness, reduced airway resistance (RI), and increased total lung capacity (TLC) and peak flow rate (PEF), with effects comparable to isoproterenol.

[0014] In a chronic obstructive pulmonary disease (COPD) model, inhaled administration of TAA nanocrystals (20 mg / kg) significantly improved ventilation function in COPD mice, manifested as increased dynamic compliance (Cydn), decreased total lung capacity (TLC), and reduced respiratory rate (RI), with effects comparable to budesonide. Except for Cydn, low-dose TAA nanocrystals (10 mg / kg) significantly reversed these changes.

[0015] TAA solid dispersion (TAA-SD) (200 mg / kg) administered orally also significantly improved ventilation function in COPD mice, manifested as increased Cydn, decreased TLC, and decreased RI. TAA-SD (100 mg / kg) significantly reduced RI. The same dose of TAA (200 mg / kg) had a better effect on TLC and RI than TAA alone.

[0016] TAA (100, 200 mg / kg) significantly reduced respiratory tract infection (RI) in asthmatic mice, increased their cyclodn (Cydn) level, and improved their ventilation. TAA was more effective than dexamethasone.

[0017] TAA inclusion complex nasal drops (200 mg / kg) significantly reduced TLC and RI, increased Cydn, and improved ventilation function in COPD mice. TAA inclusion complex nasal drops (100 mg / kg) significantly reduced RI. The effects of TAA inclusion complex nasal drops (200 mg / kg) on ​​RI and Cydn were significantly better than those of budesonide.

[0018] In a second aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of triacetylandrographolide (TAA) and pharmaceutically acceptable excipients.

[0019] The effective oral dose of the triacetylandrographolide is 50-800 mg / kg, and the effective dose by injection, nasal administration, or pulmonary inhalation is 10-100 mg / kg.

[0020] The pharmaceutically acceptable excipient is selected from at least one of diluents, binders, disintegrants, lubricants, cosolvents, surfactants, and propellants.

[0021] The dosage forms of the pharmaceutical composition include solid dosage forms, liquid dosage forms, and semi-solid dosage forms.

[0022] The solid dosage forms include powders, tablets, capsules, disintegrants, granules, or pills; the liquid dosage forms include emulsions, suspensions, syrups, oral liquids, injections or nasal drops, aerosols, or sprays; and the semi-solid dosage forms include gels or ointments.

[0023] The drug composition can be administered orally, by injection, by nasal administration, or by inhalation into the lungs.

[0024] The present invention also provides the use of the pharmaceutical composition in the preparation of medicaments for the treatment or prevention of airway obstructive diseases.

[0025] The airway obstructive diseases include asthma, chronic obstructive pulmonary disease, bronchitis, and acute lung injury.

[0026] The beneficial effects of this invention are:

[0027] 1. This invention is the first to experimentally discover that triacetylandrographolide can directly relax bronchial smooth muscle, thus providing a new candidate compound for the development of bronchodilators. This invention reveals that triacetylandrographolide (TAA) can selectively relax pre-constricted tracheal smooth muscle without significantly affecting resting tone. This characteristic indicates that it can reduce interference with normal airway physiological function while exerting its therapeutic effect, and has potentially better drug safety.

[0028] 2. This invention demonstrates that the mechanism of action of TAAs differs from commonly used β2-receptor agonists or anticholinergic drugs. TAAs exert their bronchodilatory effect through non-competitive antagonism of downstream cholinergic receptor signaling pathways. This novel mechanism provides a new approach for developing novel bronchodilators to overcome drug resistance in existing treatments. The optimized TAA formulation prepared in this invention shows superior efficacy not only compared to the active pharmaceutical ingredient but also to first-line treatments (such as budesonide) in various animal models of airway obstructive diseases (e.g., asthma, COPD), indicating promising application prospects. Attached Figure Description

[0029] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0030] Figure 1 The figure shows the effect of TAA on tracheal contraction in rats; in the figure, **p < 0.01, ***p < 0.001 compared with the solvent group; ###p < 0.001 compared with the blank group.

[0031] Figure 2 The diagram shows the effect of TAA nanocrystals on the tracheal rings of rats; in the figure, *p < 0.05, **p < 0.01, ***p < 0.001 compared with the solvent group; #p < 0.05, ##p < 0.01, ###p < 0.001 compared with the excipient group; &&p < 0.01, &&&p < 0.001 compared with the blank group.

[0032] Figure 3 The effect of TAA nanocrystals on acetylcholine-induced airway hyperresponsiveness in rats is shown in the figure; #p < 0.05, ##p < 0.01, compared with the solvent group.

[0033] Figure 4The effect of TAA nanocrystals administered to the lungs on the ventilation function of COPD mice is shown in the figure; #p < 0.05, ##p < 0.01, ###p < 0.001, compared with the control group; *p < 0.05, **p < 0.01, compared with the model group.

[0034] Figure 5 The graph shows the effect of TAA-SD gavage administration on the ventilation function of COPD mice; ##p < 0.01, ###p < 0.001, compared with the control group; *p < 0.05, **p < 0.01, ***p < 0.001, compared with the model group; &p < 0.05, &&p < 0.01, &&&p < 0.001, compared with the TAA group.

[0035] Figure 6 The effect of TAA gavage on ventilation function in asthmatic mice is shown in the figure; ###p < 0.001, compared with the control group; **p < 0.01, ***p < 0.001, compared with the model group.

[0036] Figure 7 The figure shows the effect of nasal administration of TAA inclusion complex nasal drops on ventilation function in mice with acute lung injury; ##p < 0.01, ###p < 0.001, compared with the control group; *p < 0.05, ***p < 0.001, compared with the model group; &p < 0.05, compared with the budesonide group. Detailed Implementation

[0037] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0038] Example 1

[0039] The specific steps for preparing a TAA nanocrystal suspension are as follows:

[0040] Weigh 200 mg of TAA raw material and dissolve it in 1 mL of ethanol to obtain a drug solution; weigh 100 mg of poloxamer 188 and 25 mg of soybean lecithin and dissolve them together in 50 mL of purified water as a stabilizer solution; perform high-speed shearing at 10000 rpm to rapidly inject the drug solution into the stabilizer solution, and continue shearing for 10 minutes to obtain a crude suspension; transfer the crude suspension to a high-pressure homogenizer and homogenize it 15 times at 15000 psi pressure to obtain a milky white, uniform TAA nanocrystal suspension.

[0041] Example 2

[0042] The specific steps for preparing a TAA solid dispersant (TAA-SD) are as follows:

[0043] 12 g of TAA and 24 g of PEG4000 were weighed at a ratio of 1:2 (w / w), and 360 mL of ethanol was added. The mixture was sonicated until completely dissolved to obtain a mixed solution of drug and carrier material. The solution was prepared using a spray drying method. The spray drying conditions were: nitrogen as the atomizing gas, 4% solid content, and air as the drying inlet air at a flow rate of 0.3 m³ / h. 3 The injection flow rate was 5 mL / min, the injection port temperature was 50℃, the injection flow rate was 5 mL / min, the solid content was 2%, the atomization pressure was 0.1 MPa, the dried solid was collected and passed through an 80-mesh sieve to obtain TAA-SD.

[0044] Example 3

[0045] The specific steps for preparing a TAA inclusion complex nasal drop are as follows:

[0046] Accurately weigh 2.0 g of TAA and dissolve it in 16 mL of anhydrous ethanol; accurately weigh 26.46 g of hydroxypropyl-γ-cyclodextrin (HP-γ-CD) and place it in a vial, then dissolve it in 40 mL of water; while stirring, slowly and uniformly add the TAA anhydrous ethanol solution to the HP-γ-CD aqueous solution, stir to form inclusion complex, remove and place at room temperature, remove the anhydrous ethanol under reduced pressure, filter through a 0.45 μm filter membrane, pre-freeze at -20℃, and freeze-dry to obtain the TAA / HP-γ-CD solid inclusion complex (TAA-CD).

[0047] Take 12.24 g of the above inclusion complex and dissolve it in 80 mL of ultrapure water. Then, add 0.08 g of potassium sorbate and 0.01 g of EDTA-2Na in sequence and stir thoroughly to dissolve. Adjust the pH to 5.8 with 0.1 M hydrochloric acid, autoclave, and dispense to obtain TAA inclusion complex nasal drops (TAA-CD-ND).

[0048] Example of implementation effect 1

[0049] The vasodilatory effect of TAA on isolated rat tracheal rings is as follows:

[0050] The tracheal rings of SD rats were used as the research object, and the number of rats (180 ± 20 g) and the number of tracheal rings in each experimental group were not less than 7.

[0051] Healthy SD rats aged 6-8 weeks were anesthetized with CO2. The neck was cut open from the larynx to the tracheal bifurcation. The trachea was removed and quickly transferred to a pre-cooled KH solution (containing 95% O2 and 5% CO2) with NaCl 120 mM, KCl 4.8 mM, MgSO4 1.4 mM, CaCl2 2.5 mM, KH2PO4 1.2 mM, glucose 11 mM, Na2EDTA 13.4 μM, NaHCO3 25 mM, and pH adjusted to 7.4 with NaOH). The trachea was fixed with a specimen needle. Under a dissecting microscope, the connective tissue around the trachea and any remaining blood clots in the lumen were carefully removed. The trachea was then cut into tracheal rings 3-4 mm wide.

[0052] The tracheal ring was secured using an L-shaped hook, with its other end connected to a force sensor in the data acquisition system to record the tracheal ring tension in an equal-length manner. The tracheal ring was first placed in pre-cooled KH solution, then the pre-cooled buffer solution was replaced with 37°C KH solution to adjust the tracheal ring tension to 1 g. The ring was allowed to equilibrate for 60 min, with the buffer solution changed every 15 min. After the tracheal ring tension stabilized, its activity was confirmed by stimulating it with 1 μM carbacholine.

[0053] (1) Elute carbacholine with KH solution until the tracheal ring tension reaches baseline. After stabilization for 10 min, stimulate the tracheal ring with 1 μM carbacholine. After the tracheal tension stabilizes for 30 min, add gradient concentrations of TAA (0.1, 0.3, 1, 3, 10, 30, 100 μM) and corresponding volumes of solvent (DMSO) sequentially, and observe and record the changes in tracheal ring tension.

[0054] (2) Use KH solution to elute carbacholine until the tracheal ring tension reaches the baseline. Then add TAA of gradient concentrations (0.1, 0.3, 1, 3, 10, 30, 100 μM) and corresponding volumes of solvent (DMSO) in sequence. Observe and record the changes in tracheal ring tension.

[0055] (3) Elute carbacholine with KH solution until the tracheal ring tension reaches baseline, then add 30 μM TAA and the same volume of solvent. After incubation for 30 min, add carbacholine at gradient concentrations (0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, 100 μM) sequentially, plot the dose-response curve of carbacholine tracheal dilation, and calculate the maximum tracheal ring contraction effect (E) induced by carbacholine in rats. max ) and half-maximal effective concentration (EC50) 50 ).

[0056] Throughout the experiment, the buffer solution temperature was maintained at 37°C, and a mixed gas (containing 95% O2 and 5% CO2) was continuously introduced to maintain its oxygen saturation.

[0057] The results showed that TAA (30-100 μM) could relax carbacholine-preconstricted rat tracheal rings in a concentration-dependent manner, with a maximum relaxation rate of 100.8 ± 7.3%. At a concentration of 100 μM, TAA was significantly more effective than the solvent (P < 0.01).

[0058] With the tension of the tracheal rings before the addition of TAA as 100%, the contraction rate of the tracheal rings in both the TAA group (0.1-100 μM) and the solvent group remained above 80%. Compared with the solvent group, TAA had no significant effect on the basal tension of the rat tracheal rings.

[0059] With the tension of the tracheal ring before the addition of carbacholine as 100%, the tracheal ring tension induced by carbacholine in the blank control group, the solvent (DMSO) group, and the TAA (30 μM) group was measured. max The percentages were 222.6 ± 9.9%, 205.0 ± 12.1%, and 141.1 ± 10.1%, respectively, EC 50 The concentrations were 0.15, 0.16, and 0.70 μM, respectively. Compared with the blank control group and the solvent group, TAA significantly reduced the E3 concentration of carbacholine. max ( Figure 1 ).

[0060] These results indicate that TAA has a direct bronchodilatory effect, without any spontaneous bronchodilatory or constrictive effect. Preliminary mechanistic studies suggest that TAA may exert its bronchodilatory effect through non-competitive antagonism of the cholinergic receptor pathway.

[0061] Example 2 of implementation results

[0062] The effects of TAA nanocrystals on the tracheal rings of rats are as follows:

[0063] TAA nanocrystal suspension was prepared using Example 1. The tracheal rings of SD rats were used as the research object. The number of rats (180 ± 20 g) and the number of tracheal rings in each experimental group were not less than 7.

[0064] Healthy SD rats aged 6-8 weeks were anesthetized with CO2. The neck was cut open from the larynx to the tracheal bifurcation. The trachea was removed and quickly transferred to a pre-cooled KH solution (containing 95% O2 and 5% CO2) with NaCl 120 mM, KCl 4.8 mM, MgSO4 1.4 mM, CaCl2 2.5 mM, KH2PO4 1.2 mM, glucose 11 mM, Na2EDTA 13.4 μM, NaHCO3 25 mM, and pH adjusted to 7.4 with NaOH). The trachea was fixed with a specimen needle. Under a dissecting microscope, the connective tissue around the trachea and any remaining blood clots in the lumen were carefully removed. The trachea was then cut into tracheal rings 3-4 mm wide.

[0065] The tracheal ring was secured using an L-shaped hook, with its other end connected to a force sensor in the data acquisition system to record the tracheal ring tension in an equal-length manner. The tracheal ring was first placed in pre-cooled KH solution, then the pre-cooled buffer solution was replaced with 37°C KH solution to adjust the tracheal ring tension to 1 g. The ring was allowed to equilibrate for 60 min, with the buffer solution changed every 15 min. After the tracheal ring tension stabilized, its activity was confirmed by stimulating it with 1 μM carbacholine.

[0066] (1) Elute carbacholine with KH solution until the tracheal ring tension reaches baseline. After stabilization for 10 min, stimulate the tracheal ring with 1 μM carbacholine. After the tracheal tension stabilizes for 30 min, add sequentially gradient concentrations of TAA nanocrystal suspension (0.1, 0.3, 1, 3, 10, 30, 100 μM TAA) and corresponding volumes of solvent (DMSO), and observe and record the changes in tracheal ring tension.

[0067] (2) Use KH solution to elute carbacholine until the tracheal ring tension reaches the baseline, and add sequentially gradient concentrations of TAA nanocrystal suspension (calculated as TAA, 0.1, 0.3, 1, 3, 10, 30, 100 μM), and corresponding volumes of solvent (DMSO), and observe and record the changes in tracheal ring tension.

[0068] (3) Carbacholine was eluted with KH solution until the tracheal ring tension reached baseline. Then, 30 μM TAA nanocrystal suspension (calculated as TAA) and the same volume of solvent (KH solution) and excipients were added. After incubation for 30 min, carbacholine concentrations (0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, 100 μM) were added sequentially. A dose-response curve of carbacholine dilating the trachea was plotted, and the maximum tracheal ring contraction effect (E) induced by carbacholine in rats was calculated. max ) and half-maximal effective concentration (EC50) 50 ).

[0069] The results showed that TAA nanocrystals (1-100 μM) could relax carbacholine-preconstricted rat tracheal rings in a concentration-dependent manner, with a maximum relaxation rate of 76.5 ± 6.5%. At concentrations of 30 and 100 μM, the effect of TAA nanocrystals was significantly better than that of solvents and excipients (P < 0.01).

[0070] With the tension of the tracheal ring before the addition of TAA nanocrystals as 100%, the contraction rate of the tracheal ring in each group of rats was maintained above 80%. Compared with the solvent group, TAA nanocrystals had no significant effect on the basal tension of the rat tracheal ring. Compared with the excipient group, TAA nanocrystals could significantly maintain the basal tension of the rat tracheal ring.

[0071] With the tension of the tracheal ring before the addition of carbacholine as 100%, the tracheal ring tension in rats induced by carbacholine was compared among the blank control group, solvent (KH solution) group, excipient group, and TAA nanocrystal group. max The percentages were 222.6 ± 9.9%, 207.9 ± 8.0%, 198.4 ± 9.6%, and 151.2 ± 13.2%, respectively, EC 50 The concentrations were 0.15, 0.18, 0.13, and 0.38 μM, respectively. Compared with the blank control group, solvent group, and excipient group, TAA nanocrystals significantly reduced the E3 concentration of carbacholine. max ( Figure 2 ).

[0072] The above results indicate that TAA nanocrystals have a direct bronchodilatory effect without spontaneous bronchodilatory or constrictive effects. Preliminary mechanistic studies suggest that TAA nanocrystals may exert their bronchodilatory effect through non-competitive antagonism of cholinergic receptor pathways.

[0073] Example of implementation effect 3

[0074] The bronchodilation effect of TAA suspension on airway hyperresponsive rats is as follows:

[0075] TAA nanocrystal suspension was prepared using Example 1, with 32 male SD rats (180 ± 20 g) as the research subjects.

[0076] Healthy SD rats were anesthetized by intraperitoneal injection of 1% sodium pentobarbital (0.1 mL / 100 g). The trachea was bluntly dissected, incised, and the endotracheal tube was ligated and fixed at the incision site with surgical sutures. The endotracheal tube was connected to a small animal pulmonary function testing device to measure airway resistance (RI), total lung capacity (TLC), and peak flow rate (PEF) in healthy SD rats, which were used as baseline values. Isoproterenol (ISO), TAA nanocrystal suspension (10, 20 mg / kg based on TAA), or PBS was administered via the airway to the lungs. After the rats' respiration stabilized, acetylcholine chloride was injected via the tail vein to challenge the rats, and changes in RI, TLC, and PEF were recorded. The effect of the drugs on tracheal dilation in rats was calculated and evaluated with the baseline value as 100%.

[0077] The results showed that, compared with the solvent, both TAA nanocrystals (20 mg / kg) and ISO significantly inhibited acetylcholine chloride-induced airway hyperresponsiveness, manifested as a significant decrease in RI and a significant increase in TLC and PEF. The effects of TAA nanocrystals and ISO were comparable. Figure 3 ).

[0078] Example of implementation effect 4

[0079] The efficacy evaluation of TAA nanocrystal suspension nebulized inhalation in improving ventilation function in COPD mice induced by combined cigarette smoke exposure and lipopolysaccharide infusion is as follows:

[0080] TAA nanocrystal suspension was prepared using Example 1, and 75 male KM mice (20 ± 2 g) were used in the experiment.

[0081] Before the trial began, participants were randomly divided into five groups based on their body weight: a control group, a model group, a budesonide group (0.5 mg / kg), and low- and high-dose nanocrystal administration groups (10 and 20 mg / kg, respectively, based on TAA), for a total of 5 groups (n = 15).

[0082] From day 1 to day 90, animals were exposed to an animal gas exposure device for fumigation, maintaining a total particulate concentration of 500 mg / m³. 3 The animals were fumigated once daily for 1 hour, 6 days a week. On day 91, 50 μL of lipopolysaccharide solution (20 μg / mL) was sprayed into the lungs of the model animals.

[0083] Mice in the treatment group were administered the drug via gavage once daily from day 92 to day 96. Mice in the control and model groups were administered the corresponding volume of 0.5% CMC-Na (0.1 mL / g) via gavage. The experiment ended on day 97, and lung function indicators such as TLC, RI, and Cydn were measured in KM mice.

[0084] The results showed that TAA nanocrystals (20 mg / kg) significantly increased Cydn, decreased TLC and RI, and improved ventilation function in COPD mice; except for Cydn, 10 mg / kg TAA nanocrystals improved all of the above indicators. The effects of TAA nanocrystals were comparable to those of budesonide. Figure 4 ).

[0085] Example of implementation effect 5

[0086] The efficacy evaluation of TAA-SD gavage administration in improving ventilation function in COPD mice induced by combined cigarette smoke exposure and lipopolysaccharide infusion is as follows:

[0087] The TAA-SD prepared in Example 2 was used in the experiment with male KM mice, a total of 105 mice (20 ± 2 g).

[0088] Before the trial began, participants were randomly divided into seven groups according to their body weight: control group, model group, dexamethasone group (1 mg / kg), TAA (200 mg / kg), and TAA-SD (calculated as TAA, 100 and 200 mg / kg), for a total of 7 groups (n=15).

[0089] First, a mouse COPD model was established. The treatment group was administered the drug by gavage once a day from day 92 to day 96. The control group and the model group were administered the corresponding volume of 0.5% CMC-Na (0.1 mL / g) by gavage. The experiment ended on day 97, and lung function indicators such as TLC, RI, and Cydn were measured in KM mice.

[0090] The results showed that TAA-SD (200 mg / kg) significantly reduced TLC and RI, increased Cydn, and improved ventilation function in COPD mice. TAA-SD (100 mg / kg) significantly reduced RI. TAA (200 mg / kg) significantly reduced TLC and RI, and at the same dose, TAA-SD was superior to TAA (… Figure 5 ).

[0091] Example of implementation effect 6

[0092] The efficacy evaluation of TAA administration in improving ventilation function in ovalbumin (OVA)-induced asthmatic mice is as follows:

[0093] Female Balb / c mice were used in the experiment, totaling 75 mice (20 ± 2 g).

[0094] Before the trial began, participants were randomly divided into five groups according to their body weight: a control group, a model group, a dexamethasone group (1 mg / kg), and low- and high-dose TAA administration groups (100 and 200 mg / kg), for a total of 5 groups (n = 15).

[0095] On days 0 and 7 of the experiment, mice were intraperitoneally injected with 100 μL of an OVA suspension containing 20 μg of OVA and 2 mg of aluminum hydroxide, while the control group received only 100 μL of 0.9% NaCl intraperitoneally. OVA was administered via nebulization for 30 min starting on day 14 for challenge, continuing for 7 days. Mice in the treatment group were administered the appropriate dose of dexamethasone or TAA via gavage starting on day 14, while mice in the control and model groups were administered the appropriate volume of 0.5% CMC-Na (0.1 mL / g) via gavage for 7 days. OVA was administered 1 h before challenge. The experiment ended 24 h after the last administration, and lung function indicators such as RI and Cydn were measured in Balb / c mice.

[0096] The results showed that TAA (100, 200 mg / kg) significantly reduced RI in asthmatic mice, increased their Cydn, and improved their ventilation function. TAA was superior to dexamethasone (…). Figure 6 ).

[0097] Example of implementation effect 7

[0098] The efficacy evaluation of TAA-CD-ND intranasal administration in improving ventilation function in mice with lipopolysaccharide-induced acute lung injury is as follows:

[0099] TAA-CD-ND was prepared using Example 3, and 60 male C57BL / 6J mice (20 ± 2g) were selected for the experiment.

[0100] Before the experiment began, the mice were randomly divided into normal group, model group, budesonide group, and TAA-CD-ND group (100, 200 mg / kg) according to their body weight, with 12 mice in each group, for a total of 60 mice.

[0101] Twenty-four hours before drug administration, mice in each group were used to establish an acute lung injury mouse model by intranasal instillation of lipopolysaccharide. After anesthetizing mice with isoflurane, lipopolysaccharide (10 mg / kg, 30 μL / mouse) was administered intranasally.

[0102] Con group and Mod group: 20 μL of normal saline was instilled into the nasal cavity;

[0103] Bud group: 20 μL budesonide suspension (0.5 mg / kg, 20 μL / animal) was administered via nasal instillation.

[0104] TAA-CD-ND group: 20 μL TAA-CD-ND administered via nasal instillation for 100 or 200 mg / kg, 20 μL / animal.

[0105] The animals in each group were administered the drug once in the morning and once in the afternoon each day, and were fed normally for the rest of the time, for a total of 7 days. The experiment ended on the 8th day, and the pulmonary ventilation function indicators such as TLC, RI, and Cydn of C57BL / 6J mice were measured.

[0106] The results showed that TAA-CD-ND (200 mg / kg) significantly reduced TLC and RI, increased Cydn, and improved ventilation function in mice. TAA-CD-ND (100 mg / kg) significantly reduced RI. The effects of TAA-CD-ND (200 mg / kg) on ​​RI and Cydn were significantly better than those of budesonide (…). Figure 7 ).

[0107] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Application of 3,14,19-Triacetylandrographolide (TAA) in the preparation of drugs that relax bronchial smooth muscle.

2. A pharmaceutical composition, characterized in that: The pharmaceutical composition contains the TAA as described in claim 1 and a pharmaceutically acceptable carrier.

3. The pharmaceutical composition according to claim 2, characterized in that: The pharmaceutically acceptable excipient is selected from at least one of diluents, binders, disintegrants, lubricants, solubilizers, surfactants, and propellants.

4. The pharmaceutical composition according to claim 3, characterized in that: The dosage forms of the pharmaceutical composition include solid dosage forms, liquid dosage forms, and semi-solid dosage forms.

5. The pharmaceutical composition according to claim 4, characterized in that: The drug composition can be administered orally, by injection, by nasal administration, or by inhalation into the lungs.

6. The pharmaceutical composition according to claim 4, characterized in that: When administered orally, the effective dose of TAA is 50-800 mg / kg, and when administered by injection, nasal administration, or pulmonary inhalation, the effective dose of TAA is 10-100 mg / kg.

7. Use of the pharmaceutical composition according to any one of claims 2-6 in the preparation of a medicament for treating or preventing airway obstructive diseases.

8. The application according to claim 7, characterized in that: The airway obstructive diseases include asthma, chronic obstructive pulmonary disease, bronchitis, and acute lung injury.

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