CN112546046B

CN112546046B - Application of arbidol hydrochloride in preparation of medicine for treating pulmonary fibrosis diseases

Info

Publication number: CN112546046B
Application number: CN202011473824.2A
Authority: CN
Inventors: 杨诚; 杨光; 周红刚; 张亮; 李建; 伦东超
Original assignee: Tianjin Jikun Pharmaceutical Technology Co ltd
Current assignee: Tianjin Jikun Pharmaceutical Technology Co ltd
Priority date: 2020-12-15
Filing date: 2020-12-15
Publication date: 2022-11-01
Anticipated expiration: 2040-12-15
Also published as: CN112546046A

Abstract

The arbidol hydrochloride has good efficacy on pulmonary fibrosis, has no adverse reaction, can slow down pulmonary fibrosis of mice induced by bleomycin, and provides good application prospects for treating, relieving or improving pulmonary fibrosis diseases.

Description

Application of arbidol hydrochloride in preparation of medicine for treating pulmonary fibrosis diseases

Technical Field

The invention belongs to a new application of arbidol hydrochloride, and particularly relates to an application of arbidol hydrochloride in preparation of a medicament for treating pulmonary fibrosis diseases.

Background

Along with the aggravation of the environmental pollution problem, the air quality is continuously reduced, the haze weather is increased, and the incidence of lung diseases is continuously increased. Pulmonary Fibrosis (PF) is the final clinical manifestation of a number of interstitial lung diseases with different etiologies, and is characterized by persistent damage to the alveoli, fibroblast proliferation and massive extracellular matrix (ECM) deposition, which results in different degrees of inflammation and fibrosis between the alveoli and the interstitium, and thus lung structural destruction and respiratory failure, and is also called Interstitial Lung Disease (ILD) or Diffuse Parenchymal Lung Disease (DPLD).

Idiopathic Pulmonary Fibrosis (IPF) belongs to the group of Idiopathic Interstitial Pneumonia (IIP) in the family of Interstitial Lung Diseases (ILDs). IPF is the most common and most severe chronic inflammatory interstitial lung disease of unknown etiology, with progressive dyspnea with irritating dry cough, with constant progression, median survival of about 2.8 years, 5-year survival rate of less than 50%, and patients who succumb to respiratory failure and secondary lung infection. All over the world, the incidence of IPF is reported to be rising in recent years without obvious geographical and ethnic differences, patients are mostly middle-aged and elderly people, the patients often develop the IPF in 50-70 years, and the children rarely develop the IPF. In view of the large number of clinical situations, the prevalence and incidence of IPF is difficult to estimate, with a probability of 15-250 in 100000 occurring, 34000 new cases per year, depending on the country, age, sex. At present, lung transplantation is the only treatment means capable of prolonging the survival time of patients with pulmonary fibrosis, and IPF drug therapy recommended by 'idiopathic pulmonary fibrosis diagnosis and treatment Chinese expert consensus guideline' mainly comprises pirfenidone, nintedanib, antacid drugs and N-acetylcysteine, wherein the approved drugs for effectively treating IPF only comprise pirfenidone and nintedanib. Although these drugs can slow the decline of lung function, they cannot reverse the progress of the disease, and a significant portion of patients have poor response to the treatment, and their specific pharmacological mechanisms have not been fully elucidated. Therefore, the occurrence and development mechanism of the pulmonary fibrosis is clarified, a new potential drug target is explored, and the development of the drug which is confirmed in curative effect, relatively safe and reasonable in price aiming at the pulmonary fibrosis has important social significance and medical significance.

Arbidol hydrochloride, which inhibits the fusion of influenza virus lipid membrane and host cell to block the replication of virus, has antiviral activity, is first marketed in Russia in 1993, and is used for preventing and treating influenza and other acute respiratory virus infections. At present, no report that arbidol hydrochloride can slow down pulmonary fibrosis is available. The structural formula of arbidol hydrochloride is as follows:

disclosure of Invention

In view of this, the present invention aims to provide a new use of arbidol hydrochloride, i.e., an application of arbidol hydrochloride in the preparation of a medicament for treating pulmonary fibrosis diseases.

Preferably, the pulmonary fibrosis disease is idiopathic pulmonary fibrosis. Namely, the arbidol hydrochloride is applied to the preparation of the medicine for treating idiopathic pulmonary fibrosis.

Preferably, the recommended human dosage range of the arbidol hydrochloride is 10mg/kg-20mg/kg.

Preferably, the animal experiment dosage of the arbidol hydrochloride is 100mg/kg-200mg/kg.

Preferably, the animal experiment dosage of the arbidol hydrochloride is 100mg/kg.

The invention also provides a pharmaceutical composition which comprises the arbidol hydrochloride, pharmaceutically acceptable auxiliary materials of the arbidol hydrochloride, and one or more than two of pharmaceutically acceptable salts, esters and hydrates.

Preferably, the pharmaceutical composition is selected from the group consisting of tablets, capsules, pills, suppositories, aerosols, oral liquid preparations, granules, powders, injections, syrups, medicated liquors, tinctures, lotions, films, or combinations thereof.

Preferably, the pharmaceutical composition is administered by oral administration, injection, implantation, external application, spray, inhalation, or a combination thereof.

Furthermore, the pharmaceutical composition is prepared by adopting a conventional or special preparation process.

Compared with the prior art, the invention has the following advantages:

the invention provides a new application of arbidol hydrochloride, namely an application of arbidol hydrochloride in preparing a medicament for treating pulmonary fibrosis diseases. The arbidol hydrochloride has good efficacy on pulmonary fibrosis, has no adverse reaction, can slow down the pulmonary fibrosis of mice induced by bleomycin, and provides good application prospect for treating, relieving or improving pulmonary fibrosis diseases.

Drawings

FIG. 1 is a graph showing the change in body weight of mice in each administration group;

FIG. 2 shows the lung function parameters of mice after bleomycin induction improvement;

FIG. 3 shows the reduction of collagen content in the lungs of mice induced by bleomycin for each group;

FIGS. 4A-4C show the reduction of pulmonary fibrosis area in mice after bleomycin induction in each group (FIG. 4A is the normal saline group, FIG. 4B is the bleomycin group; FIG. 4C is the arbidol hydrochloride group);

FIG. 5 is a bar graph of the proportion of pulmonary fibrosis in mice after bleomycin-reduced induction in each group.

Detailed Description

Unless defined otherwise, technical terms used in the following examples have the same meanings as commonly understood by one of ordinary skill in the art to which the present invention belongs. The test reagents used in the following examples, unless otherwise specified, were all conventional biochemical reagents; the experimental methods are all conventional methods unless otherwise specified.

The invention will be described in detail with reference to the following examples.

The embodiment is as follows: arbidol hydrochloride slowed down bleomycin-induced pulmonary fibrosis in mice.

Preparing an animal model: male C57BL/6J, wild type mice (week age 8-10 weeks), with 10% chloral hydrate concentration by mass volume at 0.5ml/100g (body weight) were administered to mice for intraperitoneal injection of anesthesia, and intratracheal invasive injection of 2U/Kg bleomycin.

The specific implementation mode is as follows: the mouse is weighed and recorded after anaesthetizing, the mouse is fixed on an operation table, the neck is disinfected by 70% alcohol, a wound with the length of about 1cm is vertically cut on the neck of the mouse by a scalpel, a micro forceps is used for separating tissues to expose an air pipe, a syringe is inserted into the air pipe from the annular gap of the cartilage of the air pipe to the centripetal end, then a bleomycin physiological saline solution with the volume corresponding to the body weight of the bleomycin physiological saline solution is slowly injected according to the measurement of 2U/kg, and the animal is immediately erected and rotated left and right to enable the liquid medicine to be uniformly distributed in the lung.

The blank control group was injected with the same volume of physiological saline (0.9%; naCl).

Grouping administration: the arbidol hydrochloride treatment means that when the bleomycin is treated for 7-14 days, 100mg/kg of arbidol hydrochloride is given to a mouse by intragastric administration every day, corresponding solvent physiological saline is used as a control, and the content of lung collagen and the fibrosis severity are detected after the bleomycin is treated for 14 days.

And (3) lung function detection: on the 14 th day of bleomycin injection, 10% chloral hydrate is used for carrying out intraperitoneal injection on an anesthetized mouse (0.5 ml/100 g), the supine position is fixed on an operation table, neck fur is cut off, the trachea is exposed, the trachea is separated in a blunt manner, an incision is cut at the position close to the head of the trachea, the trachea joint of an intubation tube is inserted into the trachea and is fixed by cotton threads, the mouse is transferred to a body drawing instrument platform, a respirator and the trachea joint are connected, and the Forced Vital Capacity (FVC) of the mouse is recorded, as shown in figure 2, the detection result shows that the FVC is increased after the administration of arbidol hydrochloride, and the drug is proved to improve the lung function of the bleomycin mouse.

Detecting the content of lung collagen: namely hydroxyproline content determination, which means that a mouse is sacrificed on the 14 th day of bleomycin injection, the right lung of the mouse is separated, the mouse is placed into a 5ml ampere bottle and is dried in a 120 ℃ oven, the pH value is adjusted to 6.5-8.0 after hydrolysis under the action of hydrochloric acid, residues are filtered, PBS is added to adjust the total volume to 10ml, 50 mul of sample is taken, 350 mul of deionized water is added, 200 mul of Chloramine T (Chloramine T) solution is added for incubation for 20 minutes at room temperature, 200 mul of perchloric acid (perchloric acid) is added for incubation for 5 minutes at room temperature, and 200 mul of P-dimethylaminobenzaldehyde (P-DMAB) is added for incubation for 20 minutes at 65 ℃. And (3) measuring the light absorption value of the sample at 570nm in a 96-well plate from 200 mu L to the standard plate, drawing a standard curve by using the standard substance reading, and further obtaining the hydroxyproline concentration Cs of the measured sample according to a formula obtained by the standard curve. The amount of hydroxyproline contained in the whole right lung was converted into W = Cs × 8 (dilution of the sample to be measured) × 10 (total volume of the sample) by the following formula.

After the bleomycin (2U/Kg) is used for treating the mice to induce the pulmonary fibrosis, 100mg/Kg of arbidol hydrochloride or a corresponding solvent (normal saline) is given to the mice by gastric lavage, the mice take lung tissues to observe the fibrosis severity after 14 days of bleomycin treatment, and the molding control reagent is the normal saline. Arbidol hydrochloride mice began a slow weight regain from dosing (see figure 1) and improved lung function compared to saline-dosed mice (see figure 2,. Beta., p <0.05, i.e. statistically significantly different). In addition, the hydroxyproline content was significantly reduced in lung tissue of arbidol hydrochloride mice (table 1, fig. 3,. X, p <0.01, i.e. statistically significant differences), indicating that arbidol hydrochloride was able to reduce bleomycin-induced collagen content. The mice lung tissue sections were H & E stained (fig. 4A-4C, scale: 100 μm) and quantitative statistics of fibrosis were performed on the lung tissue sections (fig. 5, p <0.01, i.e. statistically significant differences) and the degree and area of pulmonary fibrosis were found to be significantly lower in the arbidol hydrochloride group than in the normal saline group (table 2, fig. 5).

TABLE 1 arbidol hydrochloride inhibits bleomycin-induced hydroxyproline content (microgram/right lung)

Control group	Bleomycin group	Arbidol hydrochloride
			100.50±10.25	147.25±10.60	112.2±9.12

TABLE 2 Arbidol hydrochloride inhibition of bleomycin-induced pulmonary fibrosis (percent)

Bleomycin group	Arbidol hydrochloride
		26.26±2.35	5.61±3.74

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and should not be taken as limiting the invention, so that any modifications, equivalents, improvements and the like, which are within the spirit and principle of the present invention, should be included in the scope of the present invention.

Claims

1. The application of the arbidol hydrochloride in preparing the medicament for treating the idiopathic pulmonary fibrosis diseases is characterized in that the recommended human dosage range of the arbidol hydrochloride is 10mg/kg-20mg/kg.

2. Use according to claim 1, characterized in that: the animal experiment dosage of the arbidol hydrochloride is 100mg/kg-200mg/kg.

3. Use according to claim 1, characterized in that: the animal experiment dosage of the arbidol hydrochloride is 100mg/kg.