WO2024193322A1 - Use of risperidone in treatment of organ fibrosis - Google Patents

Abstract

Use of risperidone in the preparation of a drug for treating organ fibrosis. The synthesis of collagen is inhibited by inhibiting glycine transporter GLYT1 with risperidone, thereby significantly improving bleomycin-induced and formed pulmonary fibrosis. Risperidone has less influence on other physiological functions, good safety, and low side effects.

Description

Application of risperidone in the treatment of organ fibrosis Technical Field

The invention belongs to the field of medicine, and in particular relates to an application of risperidone in treating organ fibrosis.

Background Art

Organ fibrosis is a pathological change in which the fibrous connective tissue increases and the number of parenchymal cells decreases within organ tissues. It is a common pathological feature of many chronic diseases. Fibrosis can affect any organ. According to statistics, up to 45% of deaths are caused by fibrosis. In addition, common diseases associated with fibrosis include cirrhosis, hepatitis, non-alcoholic fatty liver disease, chronic kidney disease, myocardial infarction, heart failure, diabetes, idiopathic pulmonary fibrosis and scleroderma. Patients with fibrosis-related diseases account for about 1/20 of the number of patients with major organ diseases. Therefore, the medical burden brought by fibrosis is significant, and about 1/4 of the world's population is directly or indirectly affected by organ fibrosis.

Fibrosis has become a major economic burden on global healthcare. Therefore, discovering key therapeutic targets that are highly relevant to human fibrotic diseases and developing highly effective anti-fibrotic therapies targeting these targets are the focus of future research. However, despite substantial progress in the understanding of fibrotic pathology, there is still a lack of effective treatments.

One of the main means of treating organ fibrosis is to use large doses of glucocorticoids to reduce inflammatory responses, but there are serious side effects such as femoral head necrosis, and the effect is not good for chronic organ fibrosis caused by non-acute infection. Blocking the TGF-β pathway is an important target for the treatment of fibrotic diseases, but TGF-β signaling is widely distributed in the human body and participates in important physiological functions; in addition, drugs acting on the Wnt and Notch signaling pathways also have the problem of poor selectivity. Anti-organ fibrosis drugs can also be improved by regulating oxidative stress, lipid metabolism, MMP inhibitor enzymes, etc. Pirfenidone and nintedanib have been approved by the US FDA for the treatment of idiopathic pulmonary fibrosis. Nintedanib is a potent inhibitor of tyrosine kinase receptors such as VEGF, FGF, and PDGF. The mechanism of action of pirfenidone is not very clear, and it may exert anti-fibrotic effects by inhibiting inflammatory mediators such as TNF-α, IL-6, IL-12, and IL-8.

Currently, there is a lack of effective drug treatments for organ fibrosis, except for supportive care, anti-inflammatory treatments, and stem cell therapy.

Summary of the invention

In order to solve the above technical problems, the present invention provides a use of risperidone in treating organ fibrosis.

The technical solution adopted by the present invention is: application of risperidone in treating organ fibrosis.

Application of risperidone in the preparation of drugs for treating organ fibrosis.

Preferably, the structure of risperidone is as shown in Formula 1;

Preferably, the glycine transporter GLYT1 is inhibited by risperidone.

Preferably, risperidone or a medicament comprising risperidone is administered orally.

Preferably, the dosage is 2-6 mg/d.

Preferably, it is used for pulmonary fibrosis.

The advantages and positive effects of the present invention are: inhibiting collagen synthesis by inhibiting the glycine transporter GLYT1 through risperidone can significantly improve bleomycin-induced pulmonary fibrosis, and also significantly improve the already formed pulmonary fibrosis; risperidone is a drug that has been clinically used for many years, and its safety and side effects have been studied for many years; risperidone acts on the glycine transporter GLYT1, and has little effect on other physiological functions.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1: Changes in body weight of mice in Example 2;

Fig. 2 Changes in lung weight to body weight ratio of mice in Example 2;

FIG3 HE staining results of paraffin sections of mouse lung tissue in Example 2;

FIG4 is the Masson staining result of the paraffin sections of mouse lung tissue in Example 2;

Figure 5 Sirius Red staining results of paraffin sections of mouse lung tissue in Example 2;

Figure 6 Immunohistochemistry results of mouse lung tissue sections after 1 mg/kg bleomycin intervention in Example 2;

Figure 7 Immunohistochemistry results of lung tissue sections of mice after intervention with 2 mg/kg bleomycin in Example 2;

FIG8 Immunohistochemistry results of lung tissue sections of mice after 5 mg/kg bleomycin intervention in Example 2;

Figure 9: Changes in body weight of mice in Example 3;

Figure 10 Changes in lung weight of mice in Example 3;

FIG11 HE staining results of paraffin sections of mouse lung tissue in Example 3;

FIG12 Masson staining results of paraffin sections of mouse lung tissue in Example 3;

Figure 13 Sirius Red staining results of paraffin sections of mouse lung tissue in Example 3;

Figure 14 Immunohistochemistry results of mouse lung tissue sections after 1 mg/kg bleomycin intervention in Example 3;

Figure 15 Immunohistochemistry results of lung tissue sections of mice after 5 mg/kg bleomycin intervention in Example 3;

FIG16 Effects of bleomycin and risperidone on cell activity in Example 4;

Figure 17 Western blot results of pulmonary fibrosis related indicators after risperidone intervention at different times in Example 4;

FIG18 is the gray value analysis result of Western blot of pulmonary fibrosis related indicators in Example 4;

Figure 19 Effect of GlyT1 inhibitor on cell activity in Example 4;

Figure 20 shows the results of changes in the expression levels of pulmonary fibrosis-related indicators after intervention with different drugs in Example 4;

Figure 21 shows the grayscale value results of Western blot of pulmonary fibrosis related indicators after different drug interventions in Example 4.

DETAILED DESCRIPTION

In chronic obstructive pulmonary disease, with the deposition of type I and type III collagen in the extracellular matrix, the elasticity of the small airways gradually decreases, accelerating the progression of the disease. In addition, with the proliferation of hepatic stellate cells, type I and type III collagen gradually replace type IV collagen, and the structure of the hepatic sinusoidal capillaries gradually undergoes pathological changes. While changes in vascular structure induce portal hypertension and related diseases, they further aggravate the process of fibrosis. After different solid organ injuries, we can see that the level of collagen deposition in the matrix is strongly correlated with the process of organ fibrosis. Therefore, collagen deposition has a certain predictive function for the judgment of the degree of organ fibrosis and the level of prognosis.

The present invention discloses a method for treating organ fibrosis by using risperidone, which inhibits collagen synthesis and reduces organ collagen deposition, thereby reducing the formation of organ fibrosis. Risperidone is an atypical antipsychotic drug used to treat schizophrenia. In the present invention, risperidone can inhibit the glycine transporter GLYT1, reduce the transfer of glycine, a raw material for collagen synthesis, into cells, thereby reducing collagen synthesis and effectively reducing the formation of organ fibrosis. The structure of risperidone is shown in Formula 1;

Glycine transporters are responsible for the uptake of glycine into cells, where it can be used to synthesize proteins, neurotransmitters, and other important molecules. Recent studies suggest that glycine transporters may also play a role in the development of fibrosis, a pathological condition characterized by the excessive accumulation of scar tissue in the body. One mechanism by which GlyT1 inhibitors exert their anti-fibrotic effects is by increasing the availability of extracellular glycine, which has been shown to have anti-inflammatory and anti-fibrotic effects. GlyT1 inhibitors may also exert their effects by inhibiting the proliferation of myofibroblasts, the cells that produce scar tissue in fibrosis. Although the results of preclinical studies are promising, more research is needed to determine whether GlyT1 inhibitors are effective and safe for treating fibrosis in humans. Of note, the use of glycine transporter inhibitors in humans for the treatment of fibrosis is currently an active area of research and has not yet been approved for clinical use.

Inhibiting collagen synthesis by inhibiting the glycine transporter GLYT1 through risperidone can significantly improve bleomycin-induced pulmonary fibrosis and also significantly improve already formed pulmonary fibrosis. Risperidone has been used clinically for many years, mainly to treat schizophrenia by blocking and antagonizing dopamine D2 and 5-HT2 receptors. It has high affinity for D2, 5-HT2, adrenaline-a1 and adrenaline-a2, and histamine H1 receptors, medium and low affinity for 5-HT1A, 5-HT1C, and 5-HT1D receptors, and no affinity for cholinergic muscarinic receptors, β1 and β2 receptors. Risperidone has been used to treat schizophrenia for many years, and its safety and side effects have been studied for many years; it has little effect on other physiological functions.

The scheme of the present invention is described below in conjunction with the accompanying drawings, wherein the experimental methods without specific operating steps are all carried out in accordance with the corresponding product instructions, and the instruments, reagents, and consumables used in the examples can all be purchased from commercial companies unless otherwise specified.

Example 1: Risperidone alleviates bleomycin-induced pulmonary fibrosis in mice

1.1 Experimental animals

C57BL/6J mice, male, 8-10 weeks, weighing 24-25 g, were purchased from Beijing Huafukang Biotechnology Co., Ltd. The mice were kept in an SPF environment, with the room temperature maintained at 20-25°C, the humidity controlled at 50-60%, the mice were fed normally, the light was adjusted to a 12-h day-night cycle mode, and the mice were adaptively raised in the corresponding environment for 1 week before modeling. All animal experiments were carried out in accordance with the animal care regulations and guidelines of Tianjin Medical University and approved by the Animal Committee of Tianjin Medical University.

1.2 Experimental Materials

Bleomycin (BLM, manufacturer: GLPBIO, USA), risperidone (RIS, manufacturer: Zhejiang Huahai Pharmaceutical Co., Ltd., China), normal saline, avertin (tribromoethanol, manufacturer: Shanghai MacLean Biochemical Technology Co., Ltd., China), 1 ml syringe, 26 ^1/2 _G needle; eye ointment, medical tweezers, heating lamp, and surgical board at an angle of about 70° (from the horizontal direction).

1.3 Experimental method: Bleomycin was orally instilled into mice as follows:

1. Prepare pre-diluted bleomycin and administer 1 mg/kg, 2 mg/kg, and 5 mg/kg according to body weight.

2. Anesthetize mice with Avertin: weigh each mouse and calculate the Avertin dose (prepared concentration 1.25%, mouse dose: 0.2ml/10g); inhibit the mouse's activity by grabbing its neck hair, and use a 1mL syringe and 26 ¹ / _2G needle to intraperitoneally inject the dose calculated based on body weight. The anesthesia time is 10-40 minutes.

3. Within 5 minutes of injecting the anesthetic, the mouse will settle down and stop moving. Verify sedation by loss of righting reflex. If sedation is achieved, proceed to the next step.

4. Avoid excessive dryness of mouse eyes during the experiment.

5. Fill a sterile 200 μL pipette tip with the required volume of bleomycin or sterile PBS.

6. Place the mouse on a surgical board and suspend it by the upper incisors using a surgical thread loop. Ensure that there is adequate lighting to visualize the vocal cords.

7. Use sterile padded forceps to gently stretch the tongue to one side, toward the mandible, to see the vocal cords; then lower the tip of the pipette loaded with bleomycin to the back of the mouth and deliver the liquid through the vocal cords during inspiration. Wait for a wheeze to confirm that the liquid is delivered intratracheally. For animals in the control group, replace the bleomycin solution with an equal amount of sterile PBS.

8. Release the tongue and carefully remove the upper incisors from the suspension wire. Place the mouse under a heating lamp or pad until anesthesia recovers, usually within one hour after injection of the anesthetic.

9. Clean the forceps with an alcohol pad before and after each use.

10. Monitor mice daily until they are euthanized for analysis. Mice were gavaged with risperidone or water daily according to group.

11. When the experiment reaches an appropriate point, the mice are euthanized, their eyeballs are removed to draw blood, and then the mice are dissected to obtain samples.

Embodiment 2:

According to the method in Example 1, mice in different groups were modeled. The control group was modeled with physiological saline, and the experimental group was modeled with different concentrations of bleomycin. The next day after modeling, ddH ₂ O or RIS was administered intragastrically for 21 days. The specific experimental results are shown in Table 1.

Table 1

The dose is converted by the ratio of human and mouse body surface areas, and the dosage is calculated according to the maintenance dose of 6 mg/d for humans, referring to the equivalent dose coefficient conversion method in "Pharmacological Experimental Methodology" edited by Professor Xu Shuyun, where the dosage of mice is converted by body surface area = X mg/kg × 70kg × 0.0026/20g = 9.1X mg/kg, and the dosage of mice should be 0.79 mg/(kg*d) based on an adult with a body weight of 70 kg. In this embodiment, the intragastric dose of RIS is 0.79 mg/kg*d; ddH ₂ O is intragastrically administered at the same dose.

The weight of mice was measured before modeling, and the weight of mice was monitored every day during the experiment to observe the changes in weight, and compared with the beginning of the experiment at the end of the experiment. The results are shown in Figures 1 and 2. The results showed that the weight of mice in the BLM group showed a significant decrease in dose-dependent manner compared with the NC group (****p<0.0001), even lower than its initial weight, and the lung weight to body weight ratio increased significantly (*p<0.05), which was also dose-dependent; while the weight of the BLM+RIS group increased significantly compared with the BLM group, and the change in the lung weight to body weight ratio was significantly reduced (**p<0.01, ***p<0.001).

In order to further observe the damage of mouse lung tissue, the present embodiment conducted pathological detection on mouse lung tissue, and observed it under light microscope after HE staining. The results are shown in Figure 3. It was found that the tissue structure of NC group was Normal, clear alveolar structure. After BLM stimulation, the lung tissue structure of mice was obviously disordered, the alveolar structure disappeared, the alveolar septum became thicker, the blood vessels were obviously congested, and diffuse infiltration and infiltration of inflammatory cells were observed. The lung tissue was consolidated and fibrosis was obvious. RIS intervention was started the day after BLM modeling, and the degree of BLM-induced lung tissue was significantly reduced.

Masson staining of mouse lung tissue; Masson staining is a test method for detecting the secretion and deposition level of collagen fibers in tissues. After treatment with different dyes, the overall background of the tissue section was red, the nuclei were black, and the collagen fibers were blue. The content of collagen fibers can be evaluated according to the depth of blue and the range of involvement. The test results are shown in Figure 4. Compared with the NC group, a large amount of collagen deposition was observed in the lung tissue of the BLM group mice. This deposition was very obvious in the 1 mg/kg group, the 2 mg/kg group, and the 5 mg/kg group. It was diffusely distributed, with obvious lung consolidation and the disappearance of alveolar structure; the collagen deposition in the BLM+RIS group was not obvious, which was very different from the BLM group. The alveolar structure was well preserved, and there was no obvious lung consolidation-like change.

Sirius Red staining was performed on the mouse lung tissue; Sirius Red staining is also a test method for detecting the secretion and deposition level of collagen fibers in tissues. The alkaline groups in collagen fibers combine with the strongly acidic Sirius Red dye solution. Under an ordinary optical microscope, the cell nucleus appears blue, while the collagen appears red. Under a polarized light microscope, birefringence occurs, and type I and type III collagen fibers can be distinguished. The results are shown in Figure 5. It was found that there was no obvious collagen deposition in the lung tissue sections of the mice in the NC group, the alveolar structure was relatively clear, and slight collagen deposition was visible around the bronchi; after BLM stimulation, obvious collagen deposition was visible in the lung tissue sections of the mice, which was diffusely distributed; after RIS intervention, collagen deposition was significantly reduced, and the alveolar structure was relatively normal and clear, but slight collagen deposition could still be seen around the bronchi.

To further analyze the effect of RIS on collagen, immunohistochemical analysis was performed on mouse lung tissue sections to analyze the expression levels of type I and type III collagen. The test results are shown in Figures 6-8. The results showed that compared with the NC group, after BLM intervention, type I and type III collagen deposition in mouse lung tissue was obvious, and the expression levels were higher. After RIS treatment, the deposition of type I and type III collagen was significantly reduced compared with the BLM group, and the expression levels were significantly decreased.

Embodiment 3:

According to the method in Example 1, mice in different groups were modeled. The control group was modeled with physiological saline, and the experimental group was modeled with different concentrations of bleomycin. After 14 days of modeling, ddH ₂ O or RIS was administered intragastrically for 14 consecutive days. The specific experimental results are shown in Table 2.

Table 2

The dose is converted by the ratio of human and mouse body surface areas, and the dosage is calculated according to the maintenance dose of 6 mg/d for humans, referring to the equivalent dose coefficient conversion method in "Pharmacological Experimental Methodology" edited by Professor Xu Shuyun, where the dosage of mice is converted by body surface area = X mg/kg × 70kg × 0.0026/20g = 9.1X mg/kg, and the dosage of mice should be 0.79 mg/(kg*d) based on an adult with a body weight of 70 kg. In this embodiment, the intragastric dose of RIS is 0.79 mg/kg*d; ddH ₂ O is intragastrically administered at the same dose.

The weight of mice was measured before modeling, and the weight of mice was monitored every day during the experiment to observe the changes in weight, and compared with the beginning of the experiment at the end of the experiment. The results are shown in Figures 9 and 10. The results show that the weight of mice in the BLM group showed a significant decrease in dose-dependent manner compared with the NC group (****P<0.0001), even lower than their initial weight, and the lung weight to body weight ratio increased significantly (*P<0.05), which was also dose-dependent; while the weight of the BLM+RIS group increased significantly compared with the BLM group, and the change in the lung weight to body weight ratio was significantly reduced (**P<0.01, ***P<0.001). This change was effective regardless of whether RIS was used throughout the whole process.

In order to further observe the damage of mouse lung tissue in detail, the present embodiment performed pathological detection on mouse lung tissue, and observed under a light microscope after HE staining. The results are shown in Figure 11, and it was found that the tissue structure of the NC group was generally normal, and the alveolar structure was clear. After BLM stimulation, the structure of the mouse lung tissue was obviously disordered, the alveolar structure disappeared, the alveolar septum became thicker, the blood vessels were obviously congested, and diffuse infiltration and infiltration of inflammatory cells were observed. The lung tissue was consolidated and fibrosis was obvious. RIS intervention was given 14 days after BLM modeling, which could also weaken the BLM-induced lung tissue consolidation.

The results of Masson staining of mouse lung tissues are shown in Figure 12. Compared with the NC group, a large amount of collagen deposition was observed in the lung tissues of the BLM group mice, with obvious lung consolidation and disappearance of alveolar structure. The collagen deposition in the BLM+RIS group was not obvious, which was very different from the BLM group. The alveolar structure was well preserved and there was no obvious lung consolidation-like changes. In the bleomycin-induced collagen deposition in the lung tissue of mice, collagen was secreted in large quantities in the mouse lung tissue. RIS intervention could significantly reduce the bleomycin-induced collagen deposition in the mouse lung tissue.

Sirius Red staining was performed on the mouse lung tissue; the results are shown in Figure 13. It was found that there was no obvious collagen deposition in the lung tissue sections of the mice in the NC group, the alveolar structure was relatively clear, and slight collagen deposition was visible around the bronchi; after BLM stimulation, obvious collagen deposition was diffusely distributed in the lung tissue sections of the mice; after RIS intervention, collagen deposition was significantly reduced, and the alveolar structure was relatively normal and clear, but slight collagen deposition was still seen around the bronchi. This intervention had this effect regardless of whether it was performed the next day after modeling or 14 days later.

To further analyze the effect of RIS on collagen, immunohistochemical analysis was performed on mouse lung tissue sections to analyze the expression levels of type I and type III collagen. The test results are shown in Figures 14-15; the results showed that compared with the NC group, after BLM intervention, type I and type III collagen deposition in mouse lung tissue was obvious, and the expression level was higher; combined with the results of Example 2, whether RIS treatment was started the next day or 14 days later, the deposition of type I and type III collagen was significantly reduced compared with the BLM group, and the expression level was significantly decreased.

Example 4: Risperidone alleviates the destruction of A549 cells caused by bleomycin

Experimental materials: A549 cells; bleomycin (BLM, manufacturer: MedChemExpress, USA); risperidone (RIS, manufacturer: MedChemExpress, USA); GlyT1Inhibitor (GLYT1IN, manufacturer: MedChemExpress, USA); COL1A1 primary antibody (Aibotek Biotechnology Co., Ltd., China); COL3A1 primary antibody (Aibotek Biotechnology Co., Ltd., China); COL4A1 primary antibody (Aibotek Biotechnology Co., Ltd., China); β-actin primary antibody (Aibotek Biotechnology Co., Ltd., China); E-cadherin primary antibody (Aibotek Biotechnology Co., Ltd., China); α-SMA primary antibody (Aibotek Biotechnology Co., Ltd., China).

4.1 Cell proliferation toxicity analysis (CCK8)

The CCK-8 kit was used to detect cell proliferation and cytotoxicity. Its working principle is: in the presence of an electron coupling reagent, WST-8 (chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonylbenzene)-2H-tetrazole monosodium salt) can be reduced by dehydrogenases in mitochondria to generate a highly water-soluble orange-yellow formazan product, the depth of which is proportional to cell proliferation and inversely proportional to cytotoxicity. For the same cells, the depth of color is linearly related to the number of cells. Use an enzyme reader at a wavelength of 450nm to measure the color of the product. Determining the OD value can indirectly reflect the number of living cells.

The CCK8 experiment was used to detect the activity of A549 cells treated with different concentrations of BLM and RIS for 72 hours. In this example, BLM was used for stimulation when the A549 cell viability was about 80%, and 25μM BLM was used to stimulate the A549 cells for modeling; RIS was used for stimulation when the A549 cell viability was about 88%, and 20μM RIS was used as the drug treatment concentration of this example. The results are shown in A and B in Figure 16. The results show that the cell viability of A549 cells decreases with the increase of BLM and RIS concentrations. 10μM bleomycin stimulation can cause statistical differences, and 15μM RIS stimulation can cause statistical differences.

4.2 Construction of bleomycin-induced pulmonary fibrosis model in vitro

In order to explore the effect of RIS on alveolar epithelial cells under the destructive effect of bleomycin, this example constructed a bleomycin-induced pulmonary fibrosis model in vitro, which was divided into the following four groups: NC group (control group: treated with normal saline), BLM group (bleomycin treatment group: treated with 25μM BLM for 72h), BLM+RIS20μM 48h after group (bleomycin and risperidone treatment group: treated with 25μM BLM for 72h, 20μM RIS for 24h) and BLM+RIS20μM together group (bleomycin and risperidone treatment group: treated with 25μM BLM for 72h, 20μM RIS for 72h). Western blot was used to detect the expression levels of col1α1, col3α1, and α-SMA proteins, indicators related to pulmonary fibrosis. The results are shown in Figures 17-18. Compared with the control group, the expression levels of col1α1, col3α1, and α-SMA proteins in A549 cells were significantly increased after bleomycin stimulation, while RIS treatment for 24h and 72h could significantly downregulate the increase.

From the above results of cell and animal model studies, it can be seen that risperidone (RIS) can significantly improve organ fibrosis. From the perspective of the in vivo pulmonary fibrosis model, risperidone treatment can significantly improve collagen deposition after pulmonary fibrosis, the collagen content is significantly reduced, the lung tissue structure changes slightly, there is less consolidation, and the alveolar structure is relatively complete. In addition, the weight of mice that was severely reduced due to fibrosis has increased, and the enlarged lung weight to body weight ratio has decreased; from the perspective of the in vitro fibrosis model, risperidone treatment for different time periods can significantly change the expression of pulmonary fibrosis-related indicators, and can significantly reduce the increased expression caused by bleomycin stimulation.

Example 5: Study on the mechanism of risperidone inhibiting glycine transporter GLYT1

To further verify whether the downregulation was mediated by risperidone inhibiting the glycine transporter GLYT1, this example used a CCK8 kit to detect the expression of GlyT1 inhibitor in A549 cells at different concentrations. The activity of A549 cells after 72 hours of treatment with (GLYT1IN) is shown in Figure 19. As the concentration of GLYT1IN increases, the activity of A549 cells gradually decreases. Under the condition of 380nM, the activity of A549 cells is greater than 50%. In order to obtain more stable experimental results, 190nM GLYT1IN was selected as the treatment concentration of this study in subsequent studies.

To further verify whether the downregulation was mediated by risperidone's inhibition of the glycine transporter GLYT1, A549 cells were divided into four treatment groups: NC group (control group: treated with normal saline), BLM group (bleomycin treatment group: treated with 25 μM BLM for 72 h), BLM+RIS group (bleomycin and risperidone treatment group: treated with 25 μM BLM for 72 h, and 20 μM RIS for 72 h) and BLM+GLYT1IN group (bleomycin and risperidone treatment group: treated with 25 μM BLM for 72 h, and 190 nM GLYT1IN for 72 h).

Western blot was used to detect the expression levels of col1α1, col3α1, col4α1, E-cadherin (E-Cad), and α-SMA proteins, which are related indicators of pulmonary fibrosis. The results are shown in Figures 20-21. After BLM stimulation, the expression levels of col1α1, col3α1, col4α1, and α-SMA proteins increased significantly, and the expression level of E-cadherin protein decreased significantly. Compared with the BLM group, the expression levels of col1α1, col3α1, col4α1, and α-SMA proteins decreased significantly in the intervention group with RIS and GLYT1IN, and the expression level of E-cadherin protein increased significantly. It is speculated that RIS may improve the changes in fibrotic proteins caused by BLM by inhibiting GLYT1.

In summary, RIS may improve pulmonary fibrosis caused by BLM by inhibiting GLYT1. In the in vivo model experiment, risperidone can improve pulmonary fibrosis regardless of the duration of action; in the in vitro experiment, both short-term application of RIS and pre-intervention of RIS can significantly affect the expression levels of pulmonary fibrosis-related indicators.

The embodiments of the present invention are described in detail above, but the contents are only preferred embodiments of the present invention and cannot be considered to limit the scope of implementation of the present invention. All equivalent changes and improvements made according to the scope of application of the present invention should still fall within the scope of the patent coverage of the present invention.

Claims (7)

The use of risperidone in the treatment of organ fibrosis. Application of risperidone in the preparation of drugs for treating organ fibrosis. The use according to claim 1 or 2, characterized in that: the structure of risperidone is as shown in Formula 1;
The use according to claim 1 or 2, characterized in that the glycine transporter GLYT1 is inhibited by risperidone. The use according to claim 1 or 2, characterized in that risperidone or a drug containing risperidone is administered orally. The use according to claim 5 is characterized in that the dosage is 2-6 mg/d. The use according to claim 1 or 2, characterized in that it is used for pulmonary fibrosis.