KR20160038637A - A pharmaceutical composition for preventing or treating pulmonary fibrosis - Google Patents

Abstract

The present invention provides a pharmaceutical composition for preventing or treating pulmonary fibrosis comprising a compound of formula (I), a pharmaceutically acceptable salt or solvate thereof.

Description

[0001] The present invention relates to a pharmaceutical composition for preventing or treating pulmonary fibrosis,

The present invention relates to a pharmaceutical composition for preventing or treating pulmonary fibrosis.

Pulmonary fibrosis is a disease that causes dyspnea due to excessive attachment of connective tissue, especially collagen, to the alveolar wall. In pulmonary fibrosis, the actual lung tissue becomes hard and the volume of lung is reduced because the contraction and expansion for breathing is not free in the hardened lung tissue where it should be smooth and resilient. As the pathology progresses, In the worst case, it is a dangerous disease that can cause death due to dyspnea or respiratory failure. Pulmonary fibrosis is caused by various external harmful factors such as bacteria, viruses, contaminated air, tobacco smoke, dust, fungus, and radiation exposure. Inflammation is often unclear due to autoimmune reactions.

Pulmonary fibrosis due to radiation exposure may be caused by accidental radiation exposure, but it often occurs as a chronic side effect in radiation therapy for cancer, which reduces the cure rate after radiation therapy. Recently, the survival rate of cancer patients who have been treated with radiotherapy has been increasing with the development of radiotherapy technology, but pulmonary fibrosis, which is a side effect caused by radiation, has been raised as a serious problem of lowering the quality of life of cancer patients. Recent advances in radiotherapy equipment and software, and the evolution of radiological biology, have led to the development of radiotherapy techniques that can effectively control cancerous lesions while protecting normal tissues with radiation therapy from one to several (about five) , Cancer progression stage, cancer site, and the like.

Despite the development of such radiotherapy techniques, pulmonary adverse events such as pulmonary fibrosis, which is unavoidable in radiotherapy, are common side effects of chest radiotherapy patients. For the treatment of lung cancer, breast cancer or Hogichin lymphoma, radiation pneumonitis develops in 10-15% of patients who received radiation therapy to the chest, and develops to fibrosis, a chronic side effect after 6 months. This progression of pulmonary fibrosis is still maintained even after a period of about 2 years, which leads to lowering of pulmonary function, patient suffering and inconvenience of life (Non-Patent Document 1). Therefore, it is urgent to find and develop drugs that inhibit pulmonary fibrosis.

TGF-b (SMAD), a-SMA, and endothelin-1 have been reported to increase as pulmonary fibrosis predictive markers (Non-Patent Documents 2 and 3). In the treatment of pulmonary fibrosis, immunosuppressants are mainly used. Steroids and cytotoxic drugs can be used. Currently, a combination therapy of steroids with azathioprine or cyclophosphamide is used as a therapeutic agent for lung fibrosis caused by radiation exposure (Non-Patent Document 4). However, there is no clear evidence that such treatments improve patient survival or quality of life. So far, many fibrosis inhibitors have been tried in animal studies and small-scale patients, but none have been proven effective. Therefore, it is very urgent to find and develop pulmonary fibrosis inhibitors including pulmonary fibrosis by radiation.

Benjamin M et al., Chest. 11 (4): 1061-1076. 1997 Xie H et al., Exp Biol Med (Maywood): 238 (9): 1062-8. 2013. Andrew L et al., FASEB: 816-827. 2004 Ochoa et al., Journal of Medical Case Reports, 6: 413.2012

It is an object of the present invention to develop drugs which can be effectively used for the prevention or treatment of pulmonary fibrosis.

It is another object of the present invention to provide a method of preventing or treating pulmonary fibrosis comprising administering the drug.

One aspect of the present invention is

There is provided a pharmaceutical composition for preventing or treating pulmonary fibrosis comprising a compound of the following formula 1, a pharmaceutically acceptable salt or solvate thereof:

[Chemical Formula 1]

In the above formula (1), R ¹ is hydrogen or C _{1 -3} alkyl.

Another aspect of the present invention provides a method for the prevention or treatment of pulmonary fibrosis comprising administering to a mammal in need thereof a therapeutically effective amount of a compound of formula 1, a pharmaceutically acceptable salt or solvate thereof, .

The compound of formula (I) according to the present invention, its pharmaceutically acceptable salt or solvate thereof may be used for the treatment of pulmonary fibrosis, 3 and a-SMA, as well as increased normal pulmonary vascular endothelial cell-specific expression protein, VE-cadherin, which is decreased during pulmonary fibrosis. Accordingly, the pharmaceutical composition according to one aspect of the present invention is expected to be effectively used for prevention or treatment of pulmonary fibrosis, in particular pulmonary fibrosis induced by radiation exposure. In addition, it can be effectively used for pulmonary fibrosis, which may appear as a side effect in radiation therapy for cancer such as lung cancer, breast cancer, or Hodgkin's lymphoma, and thus it is preferable to overcome the obstacle of radiation therapy for cancer.

≪ Description of Symbols in Drawings >
No. IR: Control group (Radiotherapy group)
IR: Radiation group
IR + 2Me: Irradiation and 2-methoxyestradiol treatment group
FIG. 1 is a photograph of a human lung pulmonary vascular endothelial cell treated with 2-methoxyestradiol, irradiated with radiation, irradiated with radioactive rays, and then observed with a confocal microscope for changes in vascular endothelial cell shape and increase or decrease of pulmonary fibrosis-related protein by fluorescent staining.
FIG. 2A is a photograph showing the result of examination of collagen in the vascular endothelium region of the lung after irradiating the pulmonary site after treatment or untreated with 2-methoxyestradiol in a mouse animal model. FIG.
FIG. 2B is a graph showing the degree of expression of collagen, a molecule related to pulmonary fibrosis, by the trichrome staining method in FIG.
FIG. 3 is a graph showing changes in expression of p-SMAD, Smad 2/3 and alpha smooth muscle actin (SMMA), which are fibrotic proteins of the pulmonary vascular endothelial cell line, after irradiation with 2-methoxyestradiol treatment or post- It is the photograph which the result that I confirmed by the photograph is photographed.
FIG. 4 is a photograph showing the degree of expression of collagen in the pulmonary vascular endothelium by performing trichrome staining after irradiating the pulmonary site after 2-methoxyestradiol treatment or non-treatment in the lung in the mouse lung cancer model, The endothelial area was subjected to H & E staining, and the change in the morphology of the tissue of the endothelium was photographed.

Hereinafter, the present invention will be described in more detail.

All technical terms used in the present invention are used in the sense that they are generally understood by those of ordinary skill in the relevant field of the present invention unless otherwise defined. Also, preferred methods or samples are described in this specification, but similar or equivalent ones are also included in the scope of the present invention. The contents of all publications cited herein are hereby incorporated by reference in their entirety.

The inventors of the present invention have studied to develop a drug that can be effectively used for pulmonary fibrosis. As a result, it has been found that a compound represented by the following formula 1 represented by 2-methoxyestradiol is effective in increasing the amount of collagen deposited in the lung wall, To increase the amount of protein associated with the protein. In addition, it was confirmed that VE-cadherin, which is a normal pulmonary vascular endothelial cell-specific expression protein, which is decreased upon pulmonary fibrosis, is increased.

Accordingly, one aspect of the present invention is

There is provided a pharmaceutical composition for the prevention or treatment of pulmonary fibrosis comprising a compound of the formula 1, a pharmaceutically acceptable salt or solvate thereof:

[Chemical Formula 1]

In the above formula (1), R ¹ is hydrogen or C _{1 -3} alkyl.

According to one embodiment of the present invention, the compound of Formula 1 is 2-methoxyestradiol wherein R ₁ is methyl.

The compound of Formula 1 may be prepared using conventional knowledge known in the art of organic chemistry or may be purchased from commercial sources, for example, Xin M et al., An efficient, practical synthesis of 2-methoxyestradiol. Steroids. 2010 Jan; 75 (1): 53-6 .; Or Hou Y et al., A Short, Economical Synthesis of 2-Methoxyestradiol, an Anticancer Agent in Clinical. J Org Chem. 2009 Aug 21; 74 (16): 6362-4).

The pharmaceutically acceptable salt, or solvate thereof, of the compound of formula (1) may be suitably prepared or selected by those skilled in the art of organic chemistry using knowledge known in the art. In one embodiment, the solvate is a hydrate.

The pharmaceutically acceptable salt may exist as an acid addition salt in which the compound of formula (I) forms a salt with a free acid. The compound of formula 1 may form pharmaceutically acceptable acid addition salts according to conventional methods known in the art. As the free acid, an organic acid or an inorganic acid can be used. As the inorganic acid, hydrochloric acid, bromic acid, sulfuric acid, phosphoric acid or the like can be used. As the organic acid, citric acid, acetic acid, lactic acid, tartaric acid ), Succinic acid, valeric acid, maleic acid, fumaric acid, formic acid, propionic acid, oxalic acid, trifluoroacetic acid, benzoic acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, 4-toluenesulfonic acid, Glutamic acid, aspartic acid and the like can be used.

The pulmonary fibrosis may be any pulmonary fibrosis caused by various causes, and in one embodiment of the present invention the pulmonary fibrosis is caused by radiation exposure. Further, in one embodiment of the present invention, the pulmonary fibrosis is a side effect of radiation therapy generated by radiation to the normal tissue during radiation therapy for cancer. Radiation therapy for cancer, which may cause pulmonary fibrosis, includes, but is not limited to, radiation therapy for lung cancer, breast cancer, or hochikin lymphoma.

The compound of formula (I) reduces the increase of the amount of collagen deposited on the alveoli wall during pulmonary fibrosis and the increase of the amount of protein Paloidin, CA-9, p-SMAD2 / 3, a-SMA associated with pulmonary fibrosis cells; Cadherin, a normal pulmonary vascular endothelial cell-specific expression protein, which is decreased upon pulmonary fibrosis.

In pulmonary fibrosis by radiation, pulmonary vascular endothelial cells lose their original characteristics by radiation, and other types of cellular characterization, especially the amount of proteins associated with fibrotic cells, are observed. This can be easily distinguished by quantitative changes in endothelial cells and pulmonary fibrosis-related proteins. As a result of experiments, the present inventors have found that the expression of VE-cadherin, a vascular endothelial cell-specific expression protein that is reduced by radiation in human pulmonary vascular endothelial cells (HPAEC) when pretreated with 2-methoxyestradiol, CA-9, p-SMAD2 / 3 and a-SMA, which are associated with pulmonary fibrosis-related proteins that are increased by radiation, are significantly reduced compared to the untreated group (Example 1 And 3). In addition, the inventors of the present invention confirmed that pulmonary fibrosis symptoms such as collagen increase on the inner wall of the aorta were observed when irradiated to the lung area in an experimental animal (mouse). In the 2-methoxyestradiol administration group, collagen increase (Example 2). ≪ tb > < TABLE > In addition, the inventors of the present invention found that pulmonary fibrosis symptoms such as collagen increase were observed on the inner wall of the aorta while the size of lung cancer was reduced after the radiation therapy for the mouse lung cancer model. In the 2-methoxyestradiol administration group, And the degree of increase was remarkably decreased (Example 4). Thus, these experimental results show that the compounds of formula (1), including 2-methoxyestradiol, can significantly reduce the symptoms of pulmonary fibrosis seen upon radiation exposure or radiation therapy to cancer.

The pharmaceutical composition for preventing or treating pulmonary fibrosis according to the present invention may be formulated into a conventional pharmaceutical formulation known in the art. The pharmaceutical formulation may be formulated into any formulation including, but not limited to, an oral preparation, an injection, a suppository, a transdermal preparation, and a parenteral preparation. Preferably, the formulation is an oral preparation or an injection Can be formulated. The oral administration formulations may be prepared, for example, as a liquid, a suspension, a powder, a granule, a tablet, a capsule, a pill or an extract.

When formulating into each of the above-mentioned formulations, it may be prepared by adding a pharmaceutically acceptable carrier or an additive necessary for the preparation of each of the formulations.

One or more of the above carriers may be selected from the group consisting of a diluent, a lubricant, a binder, a disintegrant, a sweetener, a stabilizer, and an antiseptic as the carrier when the composition is formulated with an oral administration agent. Can be selected and used.

Examples of the diluent include lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol as the diluent, magnesium stearate or talc as the lubricant, polyvinyl pyrrolidone Or hydroxypropylcellulose are preferable. Examples of the disintegrant include carboxymethylcellulose calcium, sodium starch glycolate, polacrilin potassium, or crospovidone; Sweeteners include white sugar, fructose, sorbitol, or aspartame; Examples of the stabilizer include sodium carboxymethylcellulose, beta-cyclodextrin, white lead, or xanthan gum; As the preservative, methyl p-hydroxybenzoate, propyl p-hydroxybenzoate, or potassium sorbinate is preferable.

In addition to the above-mentioned ingredients, natural flavors such as plum flavor, lemon flavor, pineapple flavor, herb flavor and the like can be used as a known additive to enhance taste. Natural juice; Natural colorants such as chlorophyllin and flavonoid; Sweetness components such as fructose, honey, sugar alcohol, and sugar; Or acidic agents such as citric acid and sodium citrate may be mixed and used.

When the pharmaceutical composition according to the present invention is formulated into an injectable preparation, it may be prepared according to a conventional injection preparation method known in the art. The injectable preparation according to the present invention may be in the form of being dispersed in a sterile medium so that it can be used as it is when administered to a patient, or it may be in a form of being dispersed at a proper concentration by adding distilled water for injection for administration and then administered.

The techniques and pharmaceutically acceptable carriers, additives and the like required for such formulation are well known to those skilled in the art and are described in the Handbook of Pharmaceutical Excipients, 4th ed., Rowe et al., Eds. , American Pharmaceuticals Association (2003); Remington: the Science and Practice of Pharmacy, 20 ^th edition, Gennaro, Ed., Lippincott Williams & Wilkins (2000); ^{Remington's Pharmaceutical Sciences (19 th ed} ., 1995) may be referred to like.

The pharmaceutical composition according to the present invention may be administered in several divided doses so that the total daily dose of the compound of formula (I) is about 0.1-100 mg / kg on the basis of an adult in order to obtain the effect of preventing or treating pulmonary fibrosis. The dose may be suitably increased or decreased according to the type, cause, progress, route of administration, sex, age, body weight, etc. of pulmonary fibrosis.

Another aspect of the present invention provides a method of preventing or treating pulmonary fibrosis comprising administering to a animal a therapeutically effective amount of a compound of formula 1, a pharmaceutically acceptable salt or solvate thereof:

[Chemical Formula 1]

In the above formula (1), R ¹ is hydrogen or C _{1 -3} alkyl.

The details of the method for preventing or treating pulmonary fibrosis may be applied to the pharmaceutical composition for preventing or treating pulmonary fibrosis according to one aspect of the present invention.

The animal may be any mammal, including, but not limited to, humans, livestock, and pets.

Hereinafter, the present invention will be described in detail based on the following examples. However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Experimental Method

(1) Culture of used cell line

Human pulmonary vascular endothelial cells (HPAEC) were purchased from Promocell Company and cultured in a medium containing various growth factors necessary for vascular endothelial cells at 37 ° C and 5% CO ₂ .

(2) HPAEC Radiation on

HPAEC was plated on 3.5, 6, and 10 cm culture dishes and cultured in a 37 ° C CO ₂ incubator until 70-80% growth. After 2-methoxyestradiol was selectively pretreated according to the test group, gamma rays ) (Atomic Energy of Canada, Ltd., Canada) at a dose rate of 3.81 Gy / min. A total of 10 Gy was investigated.

(3) Cell Immunofluorescent staining

HPAEC was incubated on cover slides and then irradiated with 2-methoxyestradiol pretreatment, followed by 10% neutral formalin for one day and washed with PBS (phosphate buffered saline) before staining. Cadherin, HIF-1a (Hypoxia-inducible factor 1-alpha), SMAD, and p-cadherin diluted at a ratio of 1: 100 were pre-treated with 2% bovine serum albumin solution SMAD2 / 3, alpha SMA antibody was reacted at 4 DEG C for 16 hours. After washing with PBS, a secondary antibody with a fluorescent substance was diluted at a ratio of 1: 500 and reacted at 25 ° C for 1 hour. Nuclear staining was performed using DAPI (4 ', 6-diamidino-2-phenylindole) fluorescent substance. After washing with PBS, one drop of glycerol was added to the slide glass to attach stained cells, and then observed with a confocal microscope.

(4) Protein analysis using electrophoresis and immune response

After culturing the cells, the cells were lysed in a solution consisting of 150 mM sodium chloride, 40 mM Tris-Cl (pH 8.0) and 0.1% NP-40 in order to observe intracellular proteins. These samples were subjected to PAGE (polyacrylamide gel electrophoresis) containing SDS (sodium dodecyl sulfate) and then subjected to Western blotting. The proteins separated by electrophoresis were transferred to a nitrocellulose membrane, and then the amount of expression of the proteins was analyzed by immunoblotting method.

(5) Hematoxylin ( hematoxylin ) And Eojin ( Eosin ) Staining method (H & E staining )

Mouse tissues were fixed with 10% neutral formalin for one day and paraffin sections were made. Paraffins were stained with xylene, 95, 90, and 70% ethanol for 5 min each time to remove paraffin from the tissue for staining, and stained with hematoxylin for 1 min to stain nuclei And washed with running water for 10 minutes. The cells were then dipped into the eosin solution for 30 seconds and stained with 50, 70, 90, and 95% ethanol and xylenes. One drop of the mounting solution was then placed on the cover slide ) Were observed and observed with a microscope (Carl Zeiss Vision).

(6) For biotissue Trichrome ( Trichrome ) Staining method

Mouse tissues were fixed with 10% neutral formalin for one day and paraffin sections were made. For staining, xylene, 95, 90, 70% ethanol solution was immersed in ethanol solution for 5 minutes in order to remove paraffin around the tissue. Tissues were immersed in 0.1 M citric acid (pH 6.0) solution for antigen activation of the tissues and boiled for 20 minutes.

Then, 1 minute was added to Bouin's solution, 10 minutes to Weigert's hematoxylin, 10 minutes to Phosphotunstic / phosphomolydic acid, 5 minutes to aniline blue and 1 minute to 1% acetic acid. And covered with a cover glass. After sealing, they were observed with a confocal microscope.

Example  One: Immunofluorescent staining  At the cellular level by radiation used Pulmonary fibrosis  Development inhibition test

Human pulmonary vascular endothelial cells irradiated with 10 Gy of radiation were fixed with 10% formalin and reacted with Paloidine, CA-9, and VE-cadherin antibodies, followed by secondary immunity with a fluorescent substance. Microscopic observations showed that CA9 is red, paloides are green, and VE-cadherin is white. The nucleus was blue with DAPI fluorescence. In the 2-methoxyestradiol treatment group, the human pulmonary vascular endothelial cell line was irradiated at a concentration of 10 gy before 12 hours pretreatment at a concentration of 10 ng / ml before irradiation. The results are shown in Fig.

FIG. 1 is a photograph of a human lung pulmonary vascular endothelial cell treated with 2-methoxyestradiol, irradiated with radiation, irradiated with radioactive rays, and then observed with a confocal microscope for changes in vascular endothelial cell shape and increase or decrease of pulmonary fibrosis-related protein by fluorescent staining.

1, VE-cadherin is a protein present in the plasma membrane of the pulmonary vascular endothelial cell line, so that a photograph of the control group (No. IR), which is not irradiated, shows mainly white in the cell membrane. On the other hand, in the irradiation group (IR), the white area around the cell membrane was greatly decreased. Paloidin and CA-9 were pulmonary fibrosis-related proteins that were not stained in the control group but were identified as green and red in the irradiated group. In contrast, in the 2-methoxyestradiol treated group (IR + 2Me), the area around the cell membrane was significantly increased compared to the irradiated group, and Paloidine and CA-9 were significantly decreased.

From the results shown in Fig. 1, it can be seen that the treatment with 2-methoxyestradiol remarkably reduces pulmonary fibrosis-related symptoms, which are increased by irradiation with pulmonary vascular endothelial cell line, and remarkably lowers cell membrane damage caused by irradiation have.

Example  2: In the experimental animal model Pulmonary fibrosis  Development inhibition test

In order to observe the pulmonary fibrosis of vascular endothelial cells in pulmonary fibrosis after radiotherapy, 16 Gy of radiation was applied to the lungs of C57BL / 6 mice. After removing the lungs from the mouse, collagen, which is a protein that appears in the fibrosis of the pulmonary vascular endothelial cells, is identified by trichrome staining for the aortic cross section of lung tissue. The 2-methoxyestradiol treatment group was intraperitoneally administered at a dose of 150 mg / kg 1 hour before irradiation. The results are shown in Fig.

FIG. 2A is a photograph showing the result of examination of collagen in the vascular endothelium region of the lung after irradiating the pulmonary site after treatment or untreated with 2-methoxyestradiol in a mouse animal model. FIG.

FIG. 2B is a graph showing the degree of expression of collagen, which is a molecule related to pulmonary fibrosis in FIG. 2A, statistically by performing a trichrome staining method. In addition, increased fibrosis in the experimental group and increased fibrosis due to irradiation with radiation through the significance probability (**: P <0.01) was observed between 2-methoxyestradiol and 2-methoxyestradiol The results of this study are as follows.

According to FIG. 2, in the inner wall of the aorta of the control group (not shown), which had not been irradiated with radiation, a blue region, which is a collagen-stained region, was hardly observed, but blue was relatively increased in the inner wall of the aorta of the irradiated group . Therefore, it was confirmed that lung fibrosis progressed after irradiation of the lung part of the mouse. In contrast, in the 2-methoxyestradiol treated group (IR + 2Me), the blue region of collagen, a protein associated with pulmonary fibrosis, was significantly reduced compared to the irradiated group (IR). In FIG. 2A, the portion where the blue is observed is indicated by a white line.

From the results shown in Fig. 2, it can be seen that 2-methoxyestradiol remarkably inhibits fibrosis of pulmonary vascular endothelial cells by radiation.

Example  3: Radiation induced Pulmonary vascular endothelial cell line ( HPAEC ) Of Pulmonary fibrosis  Inhibition test of development

To examine the changes in the expression levels of the pulmonary fibrosis-related proteins of the human pulmonary vascular endothelial cell line according to the concentration of 2-methoxyestradiol (0.5 uM, 1 uM), the cultured human pulmonary vascular endothelial cell line was treated with 2-methoxyestradiol Was irradiated at a concentration of 0.5 uM or 1 uM for 12 hours and then irradiated at an intensity of 10 Gy. Western blotting was then performed using antibodies to p-SMAD, Smad 2/3 and alpha smooth muscle actin (SMA). Western blotting was performed with β-actin antibody to confirm the quantitative determination of protein equivalents. The results are shown in Fig.

FIG. 3 is a graph showing changes in expression of p-SMAD, Smad 2/3 and alpha smooth muscle actin (PMA) associated with fibrosis of pulmonary vascular endothelial cells after 2-methoxyestradiol treatment or post-treatment irradiation, It is the photograph which the result that I confirmed by the photograph is photographed.

According to Fig. 3, when 0.5 uM or 1 uM of 2-methoxyestradiol was treated, p-SMAD, Smad 2/3 and? SMA, which are related to the pulmonary fibrosis phenomenon, are relatively decreased at all concentrations compared to the untreated group Lt; RTI ID = 0.0 &gt; uM. &Lt; / RTI &gt;

Example  4: fibrosis and inhibition of vascular endothelial cells observed after irradiation

Trp53 <tm1Brn> / J and B6.129S4-Kras <tm4Tyj> / J were purchased from the Jackson laboratory in the United States and mated with the mutant strain of the p53 gene and the ras gene to obtain a spontaneous non-small cell lung cancer animal model *, Dooley AL *, Jacks T. 2009. Conditional mouse lung tumor models using adenoviral or lentiviral delivery of Cre recombinase, Nature protocols, 4 (7): 1064-1072, PMCID: PMC2757265)

16 Gy intensity radiation was applied to the lung area of the acquired lung cancer model. After the lungs were removed from the mice, the extracted lung organs were stained according to the above hematoxylin and eosin staining (H & E staining) or the above-mentioned trichrome staining method, and the shape of vascular endothelial cells was observed through a microscope. When the tissue is observed by this staining method, the nucleus is observed as blue and cytoplasm is seen as pink. In the case of 2-methoxyestradiol alone, 150 mg / kg was administered intraperitoneally. The 2-methoxyestradiol and radiation combination treatment group was intraperitoneally administered at a dose of 150 mg / kg 1 hour before irradiation.

The results are shown in Fig.

FIG. 4 is a photograph showing the degree of expression of collagen in the pulmonary vascular endothelium by performing trichrome staining after irradiating the pulmonary site after 2-methoxyestradiol treatment or non-treatment in the lung in the mouse lung cancer model, The endothelial area was subjected to H & E staining, and the change in the morphology of the tissue of the endothelium was photographed. In FIG. 4, the black dotted line represents the cancerous site of the lung organ, and the white square represents the pulmonary blood vessel site.

According to Fig. 4, it can be seen that the cancer size of the control group not irradiated with radiation is reduced in the irradiation group (IR). In addition, in the irradiated group (IR), the collagen area of the blue color near the endothelium, which shows the pulmonary fibrosis phenomenon, increased with the decrease of the cancer size. In contrast, in the 2-methoxyestradiol treated group (IR + 2Me), the blue collagen site was significantly reduced compared to the irradiated group (IR).

From the results of FIG. 4, it can be seen that 2-methoxyestradiol remarkably inhibits pulmonary vascular endothelial cell fibrosis, which appears as a side effect in radiotherapy for lung cancer treatment.

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the above-described embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (11)

A pharmaceutical composition for preventing or treating pulmonary fibrosis comprising a compound of the following formula (1), a pharmaceutically acceptable salt or solvate thereof:
[Chemical Formula 1]

In the above formula (1), R ¹ is hydrogen or C _{1 -3} alkyl. The pharmaceutical composition according to claim 1, wherein the compound of Formula 1 is 2-methoxyestradiol wherein R ¹ is hydrogen. The pharmaceutical composition of claim 1, wherein the pulmonary fibrosis is caused by radiation exposure. The pharmaceutical composition according to claim 1, wherein the pulmonary fibrosis is a side effect induced by radiation therapy. The pharmaceutical composition according to claim 1, wherein the radiation therapy is radiation therapy for lung cancer, breast cancer, or Hoken lymphoma. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is formulated with an oral administration agent or an injection. A method of preventing or treating pulmonary fibrosis comprising administering a therapeutically effective amount of a compound of formula 1, a pharmaceutically acceptable salt or solvate thereof, to an animal other than a human:
[Chemical Formula 1]

In the above formula (1), R ¹ is hydrogen or C _{1 -3} alkyl. 2. The method of claim 1 wherein the compound of Formula 1 is 2-methoxyestradiol wherein R &lt; ¹ &gt; is hydrogen. 2. The method of claim 1, wherein the pulmonary fibrosis is caused by radiation exposure. 2. The method of claim 1, wherein the pulmonary fibrosis is a side effect caused by radiation therapy. 6. The method of claim 1, wherein the radiation therapy is radiation therapy for lung cancer, breast cancer, or Hodgkin's lymphoma.

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