CN115282151B

CN115282151B - Application of lamitinuron

Info

Publication number: CN115282151B
Application number: CN202211017164.6A
Authority: CN
Inventors: 杨丽妲; 梁海海; 李天宇; 郭莹莹; 国佳钰; 贾琦
Original assignee: Heilongjiang nursing college
Current assignee: Heilongjiang nursing college
Priority date: 2022-08-23
Filing date: 2022-08-23
Publication date: 2023-06-09
Anticipated expiration: 2042-08-23
Also published as: CN115282151A

Abstract

The invention discloses an application of lamitinuron in preventing and treating pulmonary fibrosis diseases. The invention determines the regulation effect and the potential mechanism of the action of the lamitinib on the lung tissue lesion, and uses the lamitinib as a novel drug action target spot to be applied to the prevention and treatment of the lung fibrosis diseases, and discovers that the use of the lamitinib can inhibit the lung pathological change in the lung fibrosis process. The lamitinib is found to inhibit fibroblast activation, improve extracellular matrix deposition, and has a preventive and therapeutic effect on progressive pulmonary fibrosis of mice, and can be used as a medicament for preventing and treating pulmonary fibrosis.

Description

Application of lamitinuron

Technical Field

The invention relates to the technical field of lamitinron, in particular to application of lamitinron.

Background

Idiopathic pulmonary fibrosis (Idiopathic Pulmonary Fibrosis, IPF) is a chronic progressive interstitial lung disease, the etiology and pathogenesis of which are not yet defined. Lesions are primarily localized to the lungs, well developed in middle-aged and elderly men, and their lung histopathological and/or chest High Resolution CT (HRCT) characteristically manifest as interstitial pneumonia. Clinical features are progressive scarring or fibrosis limited to the interstitial spaces of the lungs, resulting in loss of lung function, symptoms including dry cough, tired dyspnea, and fatigue. In advanced stages of the disease, blue staining (cyanosis) of the skin may occur with reduced blood oxygen levels, and the distal ends of the fingers may thicken or stick. With the progress of the disease, the symptoms of pulmonary hypertension and right heart failure are also observed, and finally death is caused, and the median survival time after diagnosis of patients is only 3-5 years. Prednisone, azathioprine and N-acetylcysteine (NAC) have begun to be used to treat the symptoms associated with IPF, but without significantly increasing life expectancy. Lung transplantation is a selection of a part of patients, and can improve quality of life and life expectancy.

Melatonin belongs to indole heterocyclic compounds, and is mainly used for improving sleep quality and participating in synchronization of circadian rhythms. In addition, studies have shown that it has an anti-fibrotic effect in organs such as heart, liver, lung, kidney, etc. In earlier studies, we found that melatonin was able to exert an anti-pulmonary fibrosis effect through its receptor MT1/MT 2. Lamitinib is approved in the united states and japan as a selective melatonin receptor agonist for the treatment of insomnia characterized by difficulty in beginning sleep, while lamitinib also has anti-inflammatory, antioxidant, etc. Therefore, the invention proves the effect of the lamitinron in pulmonary fibrosis, explores the treatment effect of the lamitinron on pulmonary fibrosis, and brings hope to clinical pulmonary fibrosis patients.

Based on the above, the invention provides an application of lamitinron.

Disclosure of Invention

In view of the drawbacks of the prior art, an object of the present invention is to provide an application of lamitinib, so as to solve the problems set forth in the background art.

The invention solves the technical problems by adopting the following technical scheme:

the invention provides an application of lamitinron in preventing and treating pulmonary fibrosis diseases.

Preferably, the lamitinib inhibits pulmonary pathological changes in the course of pulmonary fibrosis.

Preferably, the lamitinib inhibits fibroblast activation, improves extracellular matrix deposition

Preferably, the lamitinib has a preventive and therapeutic effect on progressive pulmonary fibrosis of mice, and the lamitinib is a drug for preventing and treating pulmonary fibrosis.

The invention discloses an application of lamitinuron in preventing and treating pulmonary fibrosis diseases. Experiments prove that the lamitinib has an improving effect on the pulmonary fibrosis progress and a preventing and treating effect on pulmonary tissue lesions by using the lamitinib, and the new drug indications of the marketed drugs are found and can be used for preventing and treating human pulmonary fibrosis diseases.

Compared with the prior art, the invention has the following beneficial effects:

the invention determines the regulation effect and the potential mechanism of the action of the lamitinone on the lung tissue lesion, and uses the lamitinone as a novel drug action target spot to be applied to the prevention and treatment of the lung fibrosis diseases. The use of lamitinib was found to inhibit pulmonary pathological changes in the course of pulmonary fibrosis. Lamitinib was found to inhibit fibroblast activation, improving extracellular matrix deposition. The lamitinib is found to have prevention and treatment effects on the progressive pulmonary fibrosis of mice, and can be used as a medicament for preventing and treating the pulmonary fibrosis.

Drawings

FIG. 1 is a graph of pulmonary collagen production in a mouse with lamitinone inhibition of fibrosis according to the invention;

FIG. 2 is a graph of the effect of lamitinib on the expression of extracellular matrix proteins;

fig. 3 is a graph showing increased area of lamitinib inhibition of BLM induced pulmonary fibrosis.

Detailed Description

The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

The application of the lamitinib in preventing and treating pulmonary fibrosis diseases is provided.

The lamitinib of this example inhibits pulmonary pathological changes in the course of pulmonary fibrosis.

The lamitinib of this example inhibits fibroblast activation and improves extracellular matrix deposition

The lamitiron of the embodiment has the effects of preventing and treating the progressive pulmonary fibrosis of mice, and is a medicament for preventing and treating the pulmonary fibrosis.

The specific operation method for inhibiting the collagen deposition of the lung of the mouse by the lamitinron in the embodiment comprises the following steps:

establishing a pulmonary fibrosis model of a C57BL/6 mouse by adopting one-time intratracheal injection of bleomycin, selecting a healthy male C57BL/6 mouse with the age of 6-8 weeks, exposing a trachea after 1% sodium pentobarbital is anesthetized, and forming the pulmonary fibrosis model after 21 days by intratracheal injection of Bleomycin (BLM) or physiological Saline (Saline) at 5 mg/kg;

then, the lung tissues of the mice with all components are obtained, RNA and protein of the lung tissues of the mice are extracted, and the results show that the collagen deposition and extracellular matrix protein expression of the lung of the mice with the BLM group are obviously higher than those of the mice with the Saline group, and simultaneously, the differentiation of the lung fibroblast to the myofibroblast is increased;

after the conventional extraction of mouse lung tissue RNA, real-time PCR finds that the content of Collagen I and Collagen III in the mouse lung tissue is obviously increased after BLM injection, and meanwhile, the simple intratracheal instillation of lamitinone has no obvious influence on the mouse lung, but the instillation of lamitinone obviously inhibits Collagen generation in a pathological state.

The operation method for inhibiting the fibroblast from secreting extracellular matrix protein by the lamitinib of the embodiment comprises the following steps:

A. by Western blot detection, in BLM-induced mouse lung tissue, lamitinon inhibited protein expression of FN1 and LC 3; n=5; * P < 0.05vs control group;

B. isolation and culture of primary lung fibroblasts, lamitinone has an improving effect on Fn1 up-regulation caused by TGF-beta 1.

The lamitinib of the embodiment prevents and treats progressive pulmonary fibrosis of mice;

A. a model diagram of mice administered with lamitinib prophylaxis and therapy;

B. lamitinib has a preventive effect on BLM-induced progressive pulmonary fibrosis;

C. after pulmonary fibrosis is established, intraperitoneal injection of lamitinib inhibits the increase in the area of fibrosis.

In the prevention and treatment of the mice of the embodiment, the mice are intraperitoneally injected with lamitiron every other day after being instilled with BLM for 3 days in the trachea until the materials are obtained 21 days after BLM induction;

in the treatment group of mice, an idiopathic pulmonary fibrosis model is formed after BLM induction for 3 weeks, and then the intraperitoneal injection of lamitinone is started every other day until the 6 th week, and the mice are sacrificed to obtain materials;

lung tissue was collected and immediately frozen in liquid nitrogen or 4% paraformaldehyde fixed for further study. Dyeing by Masson;

after 3 weeks of intratracheal instillation of BLM, mice were given intraperitoneal injections of lamitinib every other day, effectively reducing the fibrotic area, indicating that lamitinib was able to inhibit BLM-induced changes in lung histomorphology and increases in fibrotic area.

Example 1:

lamitinib inhibits pulmonary collagen deposition in mice

The experiment adopts one-time intratracheal injection of bleomycin to establish a pulmonary fibrosis model of a C57BL/6 mouse. Healthy male C57BL/6 mice of 6-8 weeks old were selected, 1% pentobarbital sodium was anesthetized, and then the trachea was exposed, and Bleomycin (BLM) or physiological Saline (Saline) was injected into the trachea at 5mg/kg, and a pulmonary fibrosis model was formed after 21 days.

Subsequently, the mouse lung tissue of each component was sampled, and mouse lung tissue RNA and protein were extracted, and it was found that BLM group mice had significantly higher lung collagen deposition and extracellular matrix protein expression than Saline group, while the differentiation of lung fibroblasts into myofibroblasts was increased (FIGS. 1A-D).

Since myofibroblasts secrete collagen in large amounts, which is one of the characteristics of pulmonary fibrosis, we examined the regulation of collagen production by lamitinone. After the RNA of the mouse lung tissue is routinely extracted and real-time PCR (polymerase chain reaction) is used for finding that the content of Collagen I and Collagen III in the mouse lung tissue is obviously increased after BLM injection. Meanwhile, the single intratracheal instillation of lamitinib has no significant effect on the lung of the mice, but instillation of lamitinib significantly inhibits collagen production in pathological conditions (fig. 1E);

the a-c.pcr results showed an increase in mRNA levels of fibrotic mouse lung collagen (Col 1), fibronectin (Fn 1), myofibroblast markers (ACTA 2), n=5; western Blot detects that the content of Fn1 in the pulmonary fibrosis mice is significantly up-regulated, n=3; E. the intraperitoneal injection of lamitinib was able to prevent bleomycin-induced collagen deposition, n=4. * P < 0.05vs control group, < P < 0.01vs control group;

example 2: lamitinib inhibits fibroblast from secreting extracellular matrix protein

Fn1 is an extracellular matrix protein that is important in various processes such as cell adhesion, proliferation, differentiation, etc. Western blots showed that lamitinib could reverse BLM-induced increases in Fn1 protein levels (fig. 2A). Studies have shown that LC3 expression is increased in lung tissue of IPF patients, while melatonin receptor agonists are believed to have antioxidant effects, involved in regulating mitochondrial function. Based on the protein results, we found that lamitinron also has inhibitory effect on autophagy marker LC 3. (FIG. 2A)

Meanwhile, lung fibroblasts are the main effector of pulmonary fibrosis, and excessive activation of fibroblasts promotes the occurrence and development of fibrotic diseases. We isolated lung fibroblasts from 1-3 day old mice, with up-regulated Fn1 expression after TGF- β1 induction, while lamitinib reduced the protein level of Fn1 (fig. 2B). These data indicate that lamitinone can play a protective role in pulmonary fibrosis by acting on pulmonary fibroblasts to inhibit their secretion of fibronectin.

A. Lamitinib inhibited protein expression of FN1 and LC3 in BLM-induced mouse lung tissue by Western blot detection. n=5; * P < 0.05vs control group. B. Isolation and culture of primary lung fibroblasts, lamitinone has an improving effect on Fn1 up-regulation caused by TGF-beta 1.

Example 3: lamituron for preventing/treating progressive pulmonary fibrosis in mice

Since melatonin is widely believed to exert biological activity in combination with its receptor MT1/MT2, we explored whether lamitinone as a melatonin receptor agonist is also involved in the occurrence of pulmonary fibrosis. We discuss the prophylactic and therapeutic effects of lamitinron, respectively (fig. 3A). For the mice in the prevention group, the mice are intraperitoneally injected with lamitiron every other day after being instilled with BLM in the trachea for 3 days until the materials are obtained 21 days after BLM induction; in the treated mice, a idiopathic pulmonary fibrosis model was formed 3 weeks after induction with BLM, and intraperitoneally injecting lamitinon every other day was started until week 6, and the mice were sacrificed. Lung tissue was collected and immediately frozen in liquid nitrogen or 4% paraformaldehyde fixed for further study. By Masson staining we found that lamitinron itself had no effect on lung tissue morphology, but advanced intraperitoneal administration of lamitinron was able to reverse BLM-induced pulmonary fibrosis in mice (fig. 3B). Also, intraperitoneally injecting lamitiron every other day after 3 weeks of intratracheal instillation of BLM effectively reduced the area of fibrosis (fig. 3C), indicating that lamitiron was able to inhibit BLM-induced changes in lung histomorphology and increased area of fibrosis.

A. Pattern diagram of mice administered with lamitinib prophylaxis and therapy. B. Lamitinib has a preventive effect on BLM-induced progressive pulmonary fibrosis. C. After pulmonary fibrosis is established, intraperitoneal injection of lamitinib inhibits the increase in the area of fibrosis.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Furthermore, it should be understood that although the present disclosure describes embodiments, not every embodiment is provided with a separate embodiment, and that this description is provided for clarity only, and that the disclosure is not limited to the embodiments described in detail below, and that the embodiments described in the examples may be combined as appropriate to form other embodiments that will be apparent to those skilled in the art.

Claims

1. Use of lamitide in the preparation of a medicament for preventing and treating pulmonary fibrosis diseases.

2. Use of lamitiron according to claim 1 for the preparation of a medicament for the prevention and treatment of pulmonary fibrosis diseases, wherein lamitiron inhibits pulmonary pathological changes in the course of pulmonary fibrosis.

3. Use of lamitiron according to claim 1 for the preparation of a medicament for the prevention and treatment of pulmonary fibrosis diseases, wherein lamitiron inhibits fibroblast activation and improves extracellular matrix deposition.

4. The use of lamitiron according to claim 1 for the preparation of a medicament for the prevention and treatment of pulmonary fibrosis, wherein lamitiron has a preventive and therapeutic effect on progressive pulmonary fibrosis in mice, and lamitiron is a medicament for the prevention and treatment of pulmonary fibrosis.