WO2015131655A1

WO2015131655A1 - Use of fsk in preventing and treating chronic obstructive pulmonary disease

Info

Publication number: WO2015131655A1
Application number: PCT/CN2015/000010
Authority: WO
Inventors: 杨为民; 王蕾; 翁稚颖; 许云龙; 刘霞; 杜晓华; 彭沛华; 王友兰; 卿晨
Original assignee: 昆明医科大学
Priority date: 2014-03-07
Filing date: 2015-01-06
Publication date: 2015-09-11
Also published as: CN103816146A

Abstract

Provided in the invention is a novel use of the compound forskolin (FSK) in preventing and treating chronic obstructive pulmonary disease (COPD). The FSK can be independently used as an effective component for drug use or health-care use, and also can be prepared into tablets, buccal tablets, capsules, granules, oral solutions, injections, aerosols, emplastrum and the like with other components free from repulsive interactions. The present invention adopts administration methods such as FSK oral intragastric administration and intraperitoneal injections and the like, and experimental research has been performed on a cigarette smoke inhalation induction mouse COPD model and an elastase induction mouse COPD model to prove that FSK has an obvious preventative/treatment effect on the cigarette smoke inhalation induction COPD and the elastase induction COPD. Furthermore, the acute toxicity and long term toxicity tests of FSK prove that the present invention is safe, the dosage of an adult is 10-120 mg every day, and the administration methods comprise oral administration, injection and spraying.

Description

Application of Forskolin in the prevention and treatment of chronic obstructive pulmonary disease

Technical field

The invention belongs to the field of medicine for treating lung diseases, and particularly relates to the application of forskolin in the prevention and treatment of chronic obstructive pulmonary disease.

Background technique

Chronic obstructive pulmonary disease (COPD) is mainly caused by smoking, air pollution and chronic lung infections. The staged processes mainly include chronic bronchitis, emphysema and asthma. The distal part of the terminal bronchus of the COPD system (including respiratory bronchioles, alveolar ducts, alveolar sacs and alveoli) swells, accompanied by destruction of the air wall, is a lung disease characterized by airflow limitation, airflow Restriction is not completely reversible, progressive development into COPD, mainly involving the lungs, but can also cause damage to organs other than the lungs. COPD ranks 4th among the world's leading causes of death. Due to its high number of patients, high mortality and heavy social and economic burdens, COPD has become an important public health problem.

Forskolin (FSK) is the main active ingredient in the Coleus forskolin plant distributed only in the northeastern part of Yunnan Province. It belongs to the diterpenoids and has the molecular formula C ₂₂ H ₃₄ O _{7 .} The molecular weight is 410, and the structural formula is as shown in the following formula (1):

FSK has antispasmodic, expectorant, anti-inflammatory, antihypertensive, anti-platelet aggregation, and reduced intraocular pressure.

Summary of the invention

The object of the present invention is to provide a use of forskolin (FSK) in the prevention and treatment of chronic obstructive pulmonary disease (COPD), aiming at discovering the new function of FSK and actively and effectively preventing and treating COPD by FSK.

The present invention is achieved by the use of a forskolin in the prevention and treatment of chronic obstructive pulmonary disease.

Preferably, the forskolin can be prepared as a tablet, a buccal tablet, a capsule, a granule, an oral solution, an injection, an aerosol, a spray or a plaster.

The invention overcomes the deficiencies of the prior art and provides an application of FSK in the prevention and treatment of COPD. FSK can be used alone as a medicinal or health-care active ingredient, or can be distributed with other non-repulsive groups to prepare a medicament of the corresponding dosage form or The health care product can be prepared, for example, into a tablet, a buccal tablet, a capsule, a granule, an oral solution, or the like, or can be prepared into an injection, an aerosol, a patch, or the like.

The invention adopts FSK oral gavage administration and intraperitoneal injection to conduct experimental research on two animal models of mouse COPD induced by elastase and cigarette smoke inhalation, and proves that FSK has obvious COPD induced by elastase and cigarette smoke inhalation. The preventive and therapeutic effect. Among them, in the elastase-induced COPD experiment, compared with the model group, FSK (1mg/kg) group can significantly reduce the lung volume-to-body ratio of elastase-induced mouse emphysema model (P<0.01); FSK(1) The 4mg/kg group significantly reduced the mean alveolar area (MAA) of the elastase-induced emphysema model in mice (P<0.05), and significantly increased the mean alveolar number (MAN) in emphysema mice (P<0.05). ), suggesting that FSK can significantly improve the pathological damage of elastase-induced emphysema in mice; and histopathological sections of the lungs show alveolar dilatation and alveolar septal rupture in each dose group of FSK compared with the model group. Alveolar fusion and other conditions have been significantly improved. In the COPD experiment induced by cigarette smoke inhalation (3 months), FSK (administered by gavage) significantly improved the pathological damage of cigarette smoke inhaled COPD model mice; and histopathological sections of the lung showed that Compared with the model group, alveolar dilatation, alveolar septal rupture and alveolar fusion were significantly improved in each dose group of FSK. Compared with the model group, FSK (2, 8 mg/kg) group can significantly reduce serum IL. -6 content.

In addition, the FSK of the present invention demonstrates that the application is safe by acute toxicity and long-term toxicity test results. The daily dosage for adults is 10 to 120 mg, and the administration methods include oral administration, injection, and spraying.

DRAWINGS

BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a graph showing the effects of FSK on the pathological morphology of lung tissue of PPE-induced COPD mice and the effect on the average alveolar area and mean alveolar number in COPD mice in Example 2 of the present invention.

Figure 2 is a graph showing the effect of FSK on the lung volume to weight ratio of PPE-induced COPD mice in Example 2 of the present invention;

Figure 3 is a graph showing the effects of FSK on the pathological morphology of lung tissue induced by cigarette smoke inhalation in COPD mice and the mean alveolar area and mean alveolar number in COPD mice;

Figure 4 is a graph showing the effect of FSK on lung volume to weight ratio of COPD mice induced by cigarette smoke inhalation in Example 3 of the present invention;

Fig. 5 is a graph showing the effect of FSK on the IL-6 content in the serum of COPD mice induced by cigarette smoke inhalation in Example 3 of the present invention.

detailed description

The present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It is understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Example 1 Preparation of FSK

(1) Take 10Kg of dried Coleus forskohlii, and extract it with 50 liters of 95% ethanol at room temperature for 3 times. The extract is concentrated under reduced pressure to obtain ethanol extract. The extract is added in small amount. After the water was well-mixed, it was partitioned with petroleum ether, chloroform and n-butanol, and concentrated under reduced pressure to give the extracts.

(2) 120 g of chloroform extract was applied to a silica gel column, eluted with increasing proportion of petroleum ether-acetone, and the elution result was monitored by thin layer chromatography TLC to obtain the effective site CF-E.

(3) The effective part CF-E can be obtained by repeated chromatography and recrystallization to obtain the larynx sheath The pure Buddha of the flower of the flower.

(4) Add FSK to starch or the corresponding auxiliary materials, mix and granulate, press into oral preparations such as tablets or capsules, or add other auxiliary materials to prepare oral liquid, spray aerosol, plaster, etc., or add injection solvent. And other excipients are made into injections.

Example 2 Experimental study on the prevention and treatment of COPD by forskolin (FSK) in a mouse model of COPD induced by elastase (PPE)

1. Test method

Model establishment and experimental group treatment: mice were randomly divided into normal control group and model group. Porcine pancreatic elastase (PPE, 0.3U/10g body weight) was instilled into mouse nasal cavity to prepare mouse COPD model. In the normal control group, the same amount of physiological saline was instilled into the nasal cavity of the mice by the same method. Mice were randomly divided into 3 groups: COPD model control group, FSK (1, 4 mg/kg) group and aminophylline (20 mg/kg) control group. Each group of mice was intraperitoneally administered once a day, and the normal control group and the COPD model control group were given an equal volume of vehicle for 11 days.

Measurement of blood oxygen partial pressure: Blood was taken from the common carotid artery immediately, and an Abbott i-stat 300 portable blood gas analyzer (Abbott i-STAT System) was used.

Determination of left lung volume and histopathological examination of the lung: The left lung of the mouse was immersed in 10% formalin solution, and the fluid volume was measured to obtain the left lung tissue volume (V), which was fixed for more than 24 hours. Each mouse was taken from the lung tissue at the largest transverse diameter of the left lung. The paraffin-embedded sections were routinely stained with HE and the histopathological changes were observed. At the same time, two slices were observed for each mouse, and five fields were randomly selected for each slice, and the bronchus and the large and medium blood vessels were avoided during the measurement.

2, the experimental results

(1) The effect of FSK on the pathological morphology of lung tissue induced by PPE in COPD mice and the effect on mean alveolar area (MAA) and mean alveolar number (MAN) in COPD mice. The results are shown in Figure 1 and Table 1 below:

Table 1 Effect of FSK on mean alveolar area and mean alveolar number in PPE-induced COPD mice

In Table 1, the data is expressed as mean±SE, and the data statistical method is one way ANOVA method (Student-Newman-keuls test), vs Model, *p<0.05, **P<0.01.

As can be seen from Figure 1 and Table 1, compared with the model group, the FSK (1, 4 mg/kg) group significantly reduced the mean alveolar area (MAA) of the PEP-induced COPD model in mice (P < 0.05), both of which were significant. The mean alveolar number (MAN) of COPD mice was increased (P<0.05), suggesting that FSK and FSK can significantly improve the pathological damage of PPE-induced COPD model in mice.

(2) The effect of FSK on lung volume to weight ratio (V/W) of PPE-induced COPD mice. The results are shown in Figure 2 and Table 2 below:

Table 2 Effect of FSK/FSK on lung volume to weight ratio in PPE-induced COPD mice

In Table 2, the data is expressed as mean±SE, and the data statistical method is one way ANOVA. Method (Student-Newman-keuls test), vs Model, *p<0.05, **P<0.01.

As can be seen from Fig. 2 and Table 2, compared with the model group, the FSK (1 mg/kg) group can significantly reduce the lung volume to body weight ratio of the PPE-induced mouse COPD model (P<0.01), suggesting that FSK induces COPD in PPE. The COPD situation in the mouse model has a significant improvement.

(3) The effect of FSK on the blood oxygen partial pressure of PPE-induced COPD mice, the results are shown in Table 3 below:

Table 3 Effect of FSK on PES-induced blood oxygen partial pressure in COPD mice

In Table 3, the data is expressed as mean±SE, and the data statistical method is Kruskal-Wallis One Way Analysis of Variance on Rank, vs Model.

As can be seen from Table 3, compared with the model group, the FSK (1 mg/kg) group can increase the blood oxygen partial pressure of the PPE-induced COPD model, but there is no statistical significance (P>0.05).

Example 3 Experimental study on the prevention and treatment of COPD by FSK in a mouse model of COPD induced by cigarette smoke inhalation

1. Experimental method

Model establishment and experimental grouping: mice were randomly divided into normal control group and COPD model. The COPD model mice were inhaled two times a day after two months of inhalation, and were administered continuously for 30 days. During the period, cigarette smoke inhalation was continued for a total of 3 months. Normal control mice were also placed in another plexiglass box to breathe air. Modeling mice were randomly divided into 3 groups: COPD model control group, FSK (2, 8 mg/kg) group. The normal control group and the COPD model control group were given an equal volume of vehicle.

Determination of serum IL-6: blood was taken from the common carotid artery, and the specimen was centrifuged at a low temperature, and the serum supernatant was taken and stored at -80 °C. The content of IL-6 in the serum was determined by enzyme-linked immunosorbent assay (ELISA), and the specific procedure was carried out according to the reagent instructions.

2, test results

(1) The effect of FSK on the pathological morphology of lung tissue induced by cigarette smoke inhalation in COPD mice and the effect on mean alveolar area (MAA) and mean alveolar number (MAN) in COPD mice. The results are shown in Figure 3 and Table 4 below. Show:

Table 4 Effect of FSK on mean alveolar area and mean alveolar number in COPD mice induced by cigarette smoke inhalation

In Table 4, the data is expressed as mean±SE, and the data statistical method is one way ANOVA method (Student-Newman-keuls test), vs Model, *p<0.05.

As can be seen from Table 4 and Figure 3, compared with the model group, the alveolar interstitial inflammation was significantly alleviated in the ISOF group, and alveolar dilatation fusion and alveolar septal rupture were significantly improved. Compared with the model group, the ISOF (2,8mg/kg) group significantly reduced the mean alveolar area (MAA) of COPD mice (P<0.05), both of which were significantly increased. The mean alveolar number (MAN) of COPD mice was increased (P<0.05), suggesting that ISOF has a significant preventive and protective effect on the pathological damage of cigarette smoke inhaled COPD mice.

(2) The effect of FSK on mouse lung volume to weight ratio (V/W), the results are shown in Figure 4 and Table 5 below:

Table 5 Effect of FSK on lung volume to weight ratio in COPD mice induced by cigarette smoke inhalation

In Table 5, the data is expressed as mean±SE, and the data statistical method is Kruskal-Wallis One Way Analysis of Variance on Rank, vs Model, *p<0.05.

As can be seen from Table 5 and Figure 4, the FSK (8 mg/kg) group reduced lung volume to weight ratio compared with the COPD model group, but there was no statistical difference.

(3) The effect of FSK on the IL-6 content in the serum of COPD mice induced by cigarette smoke inhalation. The results are shown in Figure 5 and Table 6 below:

Table 6 Effect of FSK on the content of IL-6 in serum of COPD mice induced by cigarette smoke inhalation

In Table 6, the data is expressed as mean±SE, and the data statistical method is one way ANOVA method (Fisher LSD Method), vs Model, *p<0.05, **P<0.01, ***P<0.001.

As can be seen from Figure 5 and Table 6, the FSK (2 mg/kg) group significantly reduced serum IL-6 levels (P < 0.001) compared with the model group.

Compared with the disadvantages and deficiencies of the prior art, the present invention has the following beneficial effects: the present invention can be used alone as a medicinal or health care active ingredient, or can be distributed with other non-repulsive groups to prepare a medicament or health care of the corresponding dosage form. The product is applied to the prevention and treatment of chronic obstructive pulmonary disease, has good preventive and therapeutic effects, and is safe and non-toxic.

The above is only the preferred embodiment of the present invention, and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection of the present invention. Within the scope.

Claims

A use of forskolin in the prevention and treatment of chronic obstructive pulmonary disease.
The use of forskolin in the prevention and treatment of chronic obstructive pulmonary disease according to claim 1, wherein the forskolin is a tablet, a buccal tablet, a capsule, a granule, an oral solution, an injection , aerosol, spray or plaster.