An azole compound for use in the treatment of lung disease

Abstract

This invention discloses an azole compound for use in the treatment of lung disease. Specifically, by administering a therapeutically effective amount of an azole compound or a pharmaceutically acceptable salt thereof to a subject, it can enhance pulmonary alveolar regeneration, prevent alveolar enlargement and reduce alveolar septal thickening, thereby improving or treating chronic lung inflammation, pulmonary fibrosis-related diseases and COPD.

Description

Attorney Docket No. 5487 / 0262PWO3AN AZOLE COMPOUND FOR USE IN THE TREATMENT OF LUNG DISEASECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of the priority to: U.S. Provisional Patent Application Ser. No. 63 / 744916, filed Jan 14, 2025, entitled a method for treating the interstitial lung disease by azole compounds.FIELD OF THE INVENTION

[0002] This present invention relates to a use of well-known compound, and in particular to an azole compound for use in the treatment of lung disease.DESCRIPTION OF THE RELATED ART

[0003] Acute respiratory distress syndrome (ARDS) and acute lung injury (ALI) represent clinical syndromes resulting from direct or indirect pulmonary insults that progress through sequential exudative, proliferative, and fibrotic phases. The exudative phase appears with lung injury that typified by alveolar-capillary barrier disruption, intra-alveolar edema, hyaline membrane formation and alveolar epithelium disruption. Progression to the proliferative phase involves type II pneumocyte proliferation and their differentiation either into type I pneumocytes as normal tissue repair or myofibroblasts via epithelial-mesenchymal transition, predominantly influenced by mediators such as TGF-pi. Failure of effective alveolar repair leads to either alveolar space enlargement or fibrotic remodeling, marked by extracellular matrix deposition, septal thickening, and airspace enlargement, collectively underlying the pathology of interstitial lung diseases including pulmonary fibrosis.

[0004] Therapeutic approaches targeting the inflammatory process in ARDS have so far failed to reverse disease progression, despite inflammation serving as a normal mechanism for alveolar repair and tissue healing. In addition to persistent inflammation, imbalanced extracellular matrix metabolism and abnormal interstitial cell-cell and cell-matrix interactions contribute critically to parenchymal repair outcomes. Ineffective repair or aberrant deposition of extracellular matrix results in compromised alveolar exchange and lung elasticity, underscoring the importance of restoring normal alveolar repair and epithelial cell differentiation in managing ALI / ARDS and fibrotic lung disease.SUMMARY OF THE INVENTION

[0005] The primary objective of the present invention is to provide an azole compound for use in the treatment of lung disease, wherein by administering a therapeutically effective amount of an azole compound or a pharmaceutically acceptable salt thereof, it can effectively promote tissue repair from the alveolar damages of ARDS / ALI and therefore alleviate the pathogenesis of alveolar enlargement and pulmonary fibrosis, consequently the symptom of dyspnea. Accordingly, the invention provides an effective therapeutic approach for the treatment of the repair of alveoli,Atorney Docket No. 5487 / 0262PWO3pulmonary fibrosis, interstitial lung disease or emphysema, a specific pathological feature of chronic obstructive pulmonary disease (COPD).

[0006] To achieve the above objective, the present invention discloses an azole compound for use in the treatment of lung disease. It means that by administering to a subject in need thereof a therapeutically effective amount of an azole compound, or a pharmaceutically acceptable salt or derivative thereof, it can achieve the goal of treating the lung disease related inflammation or fibrosis.

[0007] In one embodiment of this invention, the lung disease is characterized by an increasing alveolar enlargement and parenchymal remolding including interstitial fibrosis.

[0008] In the other embodiment of this invention, the lung disease is characterized by an increase in a biomarker associated with fibrosis, and the biomarker includes a-smooth muscle actin (a-SMA), procollagen lai (Collal). and metalloproteinases (MMPs).

[0009] The lung disease is acute respiratory distress syndrome (ARDS), acute lung injury (ATI), Interstitial lung diseases including idiopathic pulmonary fibrosis (IPF).

[0010] In one embodiment of this invention, the azole compound is an inhibitor of CYP51 enzyme, which can inhibit the activity of the CYP51 enzy me to reduce pulmonary cholesterol synthesis.

[0011] Preferably, the azole compound is Benznidazole which can not only reduce the expression of at least one fibrotic biomarker: a-SMA, Collal, MMP7, and MMP9, but also reduces cholesterol synthesis in the lungs and alleviating fibrosis-related symptoms.

[0012] Preferably, the therapeutically effective amount of Benznidazole is at a concentration of 12.5 pM to 200 pM; and / or 100 mg / kg to 200 mg / kg

[0013] In another embodiment of this invention, the lung disease is related to emphysema of chronic obstructive pulmonary' disease (COPD).BRIEF DESCRIPTION OF DRAWINGS

[0014] Fig. 1 shows the results of H&E staining of human lung tissue slices cultured with different concentrations of CdCk The text shown at the upper left of each figure indicates the concentration of CdCk In the bottom right figure, the black scale bar represents 200 pm, while in the remaining figures, the black scale bar represents 50 pm.

[0015] Fig. 2 shows the results of H&E staining of human lung tissue slices treated with 20 pM CdCL as observed under a microscope. The locations indicated by triangles represent interstitial edema, while the locations indicated by arrows represent interstitial fibrosis. The black scale bar represents 50 pm.

[0016] Fig 3 shows the results of MTT assay of human lung tissue and rat lung tissue treated with CdCE.

[0017] Fig. 4 shows the results of H&E staining of human lung tissue slices cultured withAtorney Docket No. 5487 / 0262PWO3different treatments in Example 3.

[0018] Fig. 5 shows the results of H&E staining of human lung tissue slices cultured with different treatments in Example 4. The black scale bar represents 500 pm.

[0019] Fig. 6 shows the results of H&E staining of human lung tissue slices cultured with different treatments in Example 5. In the two figures on the left column, the black scale bar represents 50 pm; in the two figures on the right column, the black scale bar represents 100 pm.

[0020] FIG. 7 is a set of photomicrographs illustrating the results of H&E staining of lung tissue sections from mice of each group in Example 5; the dose range covers from 100 to 200 mg / kg.

[0021] FIG. 8 is a set of magnified photomicrographs of the stained lung tissue sections from Group 5 and Group 6 shown in FIG. 7 in Example 5.DETAILED DESCRIPTION OF THE INVENTION

[0022] The present invention relates to an azole compound or a pharmaceutically acceptable salt or derivative thereof, for use in the treatment of a lung disease. In a preferred embodiment, the treatment involves administering the azole compound in a therapeutically effective amount, wherein the administration is effective in moderating alveolar enlargement and / or enhancing pulmonary alveolar regeneration in the subject.

[0023] Specifically, the present invention provides the azole compound for use in the treatment of lung disease, wherein the compound is administered in a therapeutically effective amount to achieve a therapeutic or ameliorating effect on pulmonary fibrosis, chronic obstructive pulmonary- disease (COPD), or diseases related thereto. This therapeutic effect is associated with one or more mechanisms, including:

[0024] Suppressing the expression of genes associated with pulmonary fibrosis a-SMA, Col lai, MMPs;

[0025] Reducing alveolar septal thickening;

[0026] Enhancing pulmonary alveolar regeneration; and

[0027] Preventing or moderating alveolar enlargement.

[0028] In one embodiment, the subject is a human.

[0029] The pulmonary fibrosis-related diseases are characterized by pathological features such as increased alveolar septal thickness and enlarged alveolar spaces, or by increased expression of biomarkers associated with pulmonary fibrosis, wherein the pulmonary fibrosis-related biomarkers include a-smooth muscle actin (a-SMA), procollagen lai (COL lai), and metalloproteinases (MMPs).

[0030] For example, the pulmonary fibrosis-related diseases include acute respiratory distress syndrome (ARDS), acute lung injury (AL1), idiopathic pulmonary fibrosis (IPF), and interstitial lung diseases.Attomey Docket No. 5487 / 0262PWO3

[0031] The azole compound of the present invention has inhibitory7activity against CYP51 enzyme. Accordingly, administration of the azole compound or the pharmaceutically acceptable salt thereof to the subject is capable of inhibiting pulmonary cholesterol biosynthesis while improving alveolar tissue repair and ameliorating pulmonary fibrosis, thereby effectively improving or treating pulmonary injury.

[0032] In another specific embodiment of the present invention, the azole compound or a pharmaceutically acceptable salt or derivative thereof is provided for use in the treatment of emphysema or related diseases thereof.

[0033] Preferably, in this embodiment, the azole compound is administered in a therapeutically effective amount sufficient to achieve the therapeutic effect by moderating alveolar enlargement and / or enhancing pulmonary alveolar regeneration associated with emphysema.

[0034] In one embodiment of the invention, the azole compound is Benznidazole, and the therapeutically effective amount is from 12.5 pM to 100 pM and / or 100 to 200 mg / kg.

[0035] As used herein, the term “acute respiratory distress syndrome (ARDS)’' refers to a severe acute lung injury caused by multiple pathogenic factors, characterized by hypoxemia and diffuse alveolar damage.

[0036] As used herein, the term “acute lung injury (ALI)” refers to acute and widespread lung tissue injury, typically manifested as disruption of the alveolar-capillary barrier, pulmonary edema, and impaired gas exchange, which in severe cases progresses to acute respiratory distress syndrome.

[0037] As used herein, the term “interstitial lung disease” refers to diseases affecting the lung interstitium, including alveolar septa and peribronchial tissues, with clinical manifestations including persistent dyspnea, dry cough, and impaired lung function.

[0038] As used herein, the term “chronic obstructive pulmonary7disease (COPD)” refers to a common, preventable, and treatable disease characterized by persistent respiratory symptoms and airflow limitation that is due to airway and / or alveolar abnormalities, usually caused by significant exposure to noxious particles or gases. COPD is a collective term that encompasses and is not limited to emphysema, chronic bronchitis.

[0039] As used herein, the term “emphysema” refers to a condition of the lungs characterized by the permanent, abnormal enlargement of the air spaces distal to the terminal bronchiole, accompanied by the destruction of the alveolar walls, without obvious fibrosis.

[0040] As used herein, the term “azole compound” refers to a compound comprising an azole ring, including but not limited to imidazole and triazole structures, which are commonly used as antifungal agents.

[0041] As used herein, the term “Benznidazole” refers to a nitroimidazole drug primarily used in the treatment of Chagas disease, having the chemical name N-benzyl-2-nitro-lH-imidazole-l-Atorney Docket No. 5487 / 0262PWO3acetamide.

[0042] As used herein, the term “CYP51 enzyme'’ refers to a key enzyme in the cholesterol biosynthesis pathway and a pharmacological target of antifungal drugs.

[0043] As used herein, the term '‘treating” refers to preventing, alleviating, ameliorating, or curing a disease, symptom, or pathological condition by any pharmacological, surgical, physical, or other suitable means.

[0044] As used herein, the term “effective amount” refers to an amount sufficient to produce a desired physiological or pharmacological effect in a subject. Such effect may include, but is not limited to, prevention, treatment, alleviation, amelioration, or control of a disease or symptom. The effective amount may vary depending on factors such as the subject’s age, weight, sex, health condition, disease severity, route of administration, and properties of the compound.

[0045] The following examples are provided to illustrate the technical features of the present invention and the effects that can be achieved thereby, with reference to the accompanying figures for detailed description.

[0046] Example 1 : Establishment of an Ex Vivo Interstitial Lung Disease Model

[0047] Obtained from human lung tissues, approximately 1.0-1.5 mm in thickness, were cultured in vitro for a total of 5 days. Lung slices were maintained without treatment for 2 days and d were further cultured for 3 days in complete medium containing CdCL at concentrations of 0, 20, 40, or 80 pM. respectively. The culture conditions include 37°C and 5% CO2 environment.

[0048] After 5 days of culture, the tissue slices from each group were subjected to H&E staining, as shown in Figures 1 and 2. The results in Figures 1 and 2 shows that CdCL induced pathological features of interstitial lung disease in human lung tissues, including inflammation, alveolar enlargement, and alveolar wall fibrosis. Therefore, CdCL successfully induced an interstitial lung disease cell model in human lung tissue.

[0049] Example 2: Toxicity Test

[0050] Rat lung tissue slices and human lung tissue slices were subjected to toxicity testing according to the MTT assay procedure, as shown in Figure 3.

[0051] The results indicate that, compared with human lung tissue slices, CdCL exhibits high toxicity to rat lung tissue. Human lung tissue slices are more resistant to cadmium, but cell viability decreases with increasing cadmium concentration. In addition, treatment of human lung tissue slices with 30 pM CdCL for 3 days resulted in approximately 25% relative lung injury compared to untreated slices.

[0052] These results confirm that the interstitial lung disease cell model established using CdCL-treated human lung tissue slices can serve as a reliable model for studying human pulmonary injur}’ and fibrosis.Atorney Docket No. 5487 / 0262PWO3

[0053] Example 3: Ex vivo Efficacy Experiment (I)

[0054] Human lung tissue slices were randomly divided into four groups and acclimated for 2 days. From day 3 to day 7 of culture, 30 pM CdCh was added to the medium for all groups. From day 5 to day 7, the following treatments were applied:

[0055] Group 1: No drug added.

[0056] Group 2: 12.5 pM Benznidazole added.

[0057] Group 3: 25 pM Benznidazole added.

[0058] Group 4: 0.0125% (v / v) DMSO added.

[0059] The concentration of DMSO in Group 4 corresponds to that used in the preparation of 25 pM Benznidazole.

[0060] After completion of the experiment, tissue slices from each group were subjected to H&E staining, as shown in Figure 4. The results indicate that, compared with Groups 1 and 4, Groups 2 and 3 showed no significant alveolar enlargement or alveolar damage. It demonstrates that Benznidazole of the present invention effectively improves or treats the interstitial lung disease and related complications.

[0061] Example 4: Ex vivo Efficacy Experiment (II)

[0062] Human lung tissue slices were randomly divided into six groups and acclimated for 2 days. The following culture conditions were applied:

[0063] Group 1: No drug from day 2 to day 7; harvested on day 7.

[0064] Group 2: 30 pM CdCE from day 2 to day 7; harvested on day 7.

[0065] Group 3: 30 pM CdCE from day 2 to day 5, then 30 pM CdCL with DMSO (0.025%) from day 5 to day 7; harvested on day 7.

[0066] Group 4: 30 pM CdCh from day 2 to day 5, then 30 pM CdCh with 50 pM Benznidazole from day 5 to day 7; harvested on day 7.

[0067] Group 5: 30 pM CdCh from day 2 to day 5, then 30 pM CdCE with 0.025% (v / v) DMSO from day 5 to day 7, followed by 0.025% DMSO alone from day 7 to day 9; harvested on day 9.

[0068] Group 6: 30 pM CdCh from day 2 to day 5, then 30 pM CdCh with 50 pM Benznidazole from day 5 to day 7, followed by 50 pM Benznidazole alone from day 7 to day 9; harvested on day 9.

[0069] The 0.025% (v / v) DMSO corresponds to 50 pM Benznidazole.

[0070] H&E staining was performed on all tissue slices, as show n in Figure 5.

[0071] According to the results of Figure 5, it shows that alveolar enlargement and damage were alleviated in Groups 4 and 6 compared to Groups 3 and 5, so that it indicates that Benznidazole effectively ameliorates lung injury and related complications.Atorney Docket No. 5487 / 0262PWO3

[0072] Example 5: Ex vivo Efficacy Experiment (III)

[0073] Human lung tissue slices were randomly divided into two groups and acclimated for 2 days. The following treatments were applied:

[0074] TCD group: 30 pM CdCE and 2 ng / ml TGF-pi from day 2 to day 5, followed by 2 ng / ml TGF-pi with 0.05% (v / v) DMSO from day 5 to day 9.

[0075] TCB group: 30 pM CdCb and 2 ng / ml TGF-pi from day 2 to day 5, followed by 50 pM Benznidazole with 0.05% (v / v) DMSO from day 5 to day 9.

[0076] The 0.05% (v / v) DMSO corresponds to 100 pM Benznidazole.

[0077] After 9 days of culture, tissue slices were collected and subjected to H&E staining, as shown in Figure 6.

[0078] The results of Figure 6 show that the TCD group exhibited increased alveolar septal thickness and alveolar enlargement, indicating that TGF-pi exacerbates pulmonary fibrosis. In contrast, the TCB group showed significantly reduced alveolar septal thickness and alveolar enlargement.

[0079] These results of Figure 6 indicate that administration of an effective amount of Benznidazole of the present invention to patients with interstitial lung disease or pulmonary fibrosis can effectively ameliorate lung injury, thereby achieving a therapeutic effect on acute lung injury or interstitial lung disease.

[0080] Example 6: Ex Vivo Experiment (IV)

[0081] Human lung tissue slices were acclimated for 2 days and then cultured under the conditions listed in Table 1. The 0.05% (v / v) DMSO corresponds to 100 pM Benznidazole.

[0082] Table 1 : Culture treatment conditions for each group>>>Atorney Docket No. 5487 / 0262PWO3

[0083] After the culture period, RNA was extracted from tissue slices of each group. Realtime PCR method was performed to detect the expression levels of pulmonary fibrosis-related genes in lung tissue, including a-smooth muscle actin (a-SMA), procollagen lai (COL lai), metalloproteinase 1 (MMP1), metalloproteinase 2 (MMP2), metalloproteinase 7 (MMP7), and metalloproteinase 9 (MMP9), and then analyze the data. The results are shown in Tables 2-7.

[0084] Table 2:a-SMA Relative Expression

[0085] Table 3: COLlal of Relative Expression

[0086] Table : MMP1 of Relative Expression

[0087] Table 5: MMP2 of Relative ExpressionAtorney Docket No. 5487 / 0262PWO3

[0088] Table 6: MMP7 of Relative Expression

[0089] Table 7: MMP9 of Relative Expression

[0090] The results from Tables 2 to 7 demonstrate that Benznidazole of the present invention can reduce the expression of pulmonary fibrosis-related genes, including a-SMA, COLlal, MMP7, and MMP9. These results indicate that Benznidazole can effectively achieve therapeutic or ameliorative effects on pulmonary fibrosis or related diseases by inhibiting the expression of fibrosis-related genes.

[0091] Example 7: Animal Experiment

[0092] C57BL / 6 mice were randomly divided into six groups. The experiment was conducted for a total of 11 days, with treatments administered under the following conditions:

[0093] Group 1 : No drug administration, no LPS-induced inflammation.

[0094] Group 2: LPS-induced inflammation.

[0095] Group 3: LPS-induced inflammation, administered Benznidazole 100 mg / kg.

[0096] Group 4: LPS-induced inflammation, administered 20% DMSO (equal volume 100 mg / kg Benznidazole).

[0097] Group 5: LPS-induced inflammation, administered Benznidazole 200 mg / kg.

[0098] Group 6: LPS-induced inflammation, administered 40% DMSO (equal volume 200 mg / kg Benznidazole)..

[0099] For Groups 2 to 6, LPS was delivered on Day 0 at a single dose of 5 mg / kg bodyAttorney Docket No. 5487 / 0262PWO3weight through Intraperitoneal injection. The oral administration of Benznidazole or DMSO for Groups 3 to 6 was delivered from Day 7 to Day 11 of the experiment, twice daily.

[0100] After the experiment was concluded, the lungs of the mice from each group were collected, sectioned, and subjected to H&E staining. The results are shown in Figure 7 and Figure 8.

[0101] As shown by the results in Figure 7 and Figure 8, comparison of the lung tissue sections between Group 1 and Group 2 mice shows that LPS successfully induced obvious alveolar septal destruction, interstitial tissue thickening, and significant cellular infiltration in the lungs. This means that an animal model of chronic alveolar infiltration and chronic inflammation was successfully established using LPS, and this disease model is consistent with the chronic inflammatory response of emphysema.

[0102] Furthermore, the sections from Group 4 and Group 6 show that DMSO is unable to alleviate or improve the condition of pulmonary inflammation and alveolar space enlargement.

[0103] Compared to the mice in Group 2, the lung sections of the mice in Group 3 and Group 5 clearly improved the phenomenon of pulmonary inflammation and infiltration, and w ere able to effectively reduce the size of the alveolar space. Moreover, the therapeutic effect increased with the increasing administered dose of Benznidazole.

[0104] The results shown in Figure 7 and Figure 8 demonstrate that the Benznidazole disclosed in the present invention is indeed capable of treating or improving the condition of pulmonary inflammation and infiltration, and is able to repair the lungs and enhance pulmonary alveolar regeneration. This means that by administering the Benznidazole and its related compounds disclosed in the present invention, the efficacy of effectively treating, preventing, or ameliorating, emphysema / COPD, and other related diseases characterized by destructive or inflammatory changes in the lung parenchyma can be achieved.

Claims

Attorney Docket No. 5487 / 0262PWO3WHAT IS CLAIMED IS1. An azole compound or a pharmaceutically acceptable salt or derivative thereof for use in a method of treating a lung disease in a subject in need thereof.

2. The azole compound for use according to claim 1, wherein the azole compound is Benznidazole.

3. The azole compound for use according to claim 1 , wherein the azole compound is an inhibitor of CYP51 enzyme.

4. The azole compound for use according to claim 2, wherein the Benznidazole inhibits the activity of the CYP51 enzyme.

5. The azole compound for use according to claim 1, wherein the lung disease is characterized by an increasing parenchymal remolding including interstitial fibrosis and alveolar enlargement.

6. The azole compound for use according to claim 5, wherein the lung disease is acute respiratory distress syndrome (ARDS) or acute lung injury' (ALI).

7. The azole compound for use according to claim 1, wherein the lung disease is Interstitial lung diseases.

8. The azole compound for use according to claim 7, wherein the lung disease is idiopathic pulmonary fibrosis (IPF).

9. The azole compound for use according to claim 8, wherein the pulmonary fibrosis is characterized by an increase in a biomarker associated with fibrosis, and the biomarker includes a-smooth muscle actin (a-SMA), procollagen lai (Collal), and metalloproteinases (MMPs).

10. The azole compound for use according to claim 3, wherein the azole compound reduces cholesterol synthesis in the lungs and alleviating fibrosis-related symptoms.

11. The azole compound for use according to claim 4, wherein the Benznidazole reduces cholesterol synthesis in the lungs and alleviating fibrosis-related symptoms.

12. The azole compound for use according to claim 2, wherein the therapeutically effective amount of Benznidazole is at a concentration of 12.5 pM to 100 pM.

13. The azole compound for use according to claim 2, wherein the Benznidazole leads to a reduction in alveolar septa thickness and enlargement of the alveolar space.

14. The azole compound for use according to claim 2, wherein the Benznidazole reduces the expression of at least one fibrotic biomarker: a-SMA, Collal, MMP7, and MMP9.

15. The azole compound for use according to claim 1, wherein the lung disease is chronic obstructive pulmonary disease (COPD).

16. The azole compound for use according to claim 15, wherein the lung disease is emphysema.

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