Pharmaceutical composition for prevention or treatment of systemic sclerosis

Abstract

To provide a pharmaceutical composition useful for the prevention or treatment of systemic sclerosis associated interstitial lung disease (SSc-ILD).SOLUTION: A pharmaceutical composition comprising a compound represented by the following formula or a pharmaceutically acceptable salt thereof is provided.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention provides a pharmaceutical composition for preventing or treating systemic sclerosis. [Background technology]

[0002] Systemic sclerosis (SSc) is a degenerative autoimmune rheumatic disease that causes thickening and hardening of the skin, blood vessels, and internal organs. It is a chronic disease that progresses over a long period of time. While the cause of systemic sclerosis remains unknown, there are reports that exposure to certain chemicals (toluene, benzene, vinyl chloride, silica, etc.) may also be associated with the onset of the disease. Diagnosis is based on a comprehensive medical history, physical examination, tissue tests, and blood tests. The extent of internal organ damage can be confirmed through upper and lower gastrointestinal examinations, X-rays, pulmonary function tests, electrocardiograms, and cardiac ultrasounds.

[0003] Although systemic sclerosis generally has a favorable prognosis, it can become life-threatening if it spreads to the lungs, kidneys, or heart. Interstitial lung disease and pulmonary arterial hypertension can progress, causing respiratory distress and severe hypertensive renal crisis.

[0004] Systemic sclerosis (SSc) is an autoimmune disease caused by an internal immune system error. SSc is treated by rheumatology departments to improve the various symptoms caused by autoimmune inhibition. SSc is broadly classified as limited or diffuse cutaneous, depending on the extent of skin involvement. Some SSc patients also develop symptoms of systemic sclerosis-associated pulmonary disease (SSc-ILD) along with immune dysregulation. SSc-ILD is more common in diffuse SSc patients. Furthermore, 25–30% of patients develop progressive SSc-ILD, which is known to be the leading cause of death in SSc patients (Lancet, 2017;390:1685–1699).

[0005] Systemic sclerosis-related lung disease (SSC-related lung disease) is a respiratory disease that occurs in patients with systemic sclerosis. It is diagnosed by respiratory medicine and treated to improve lung function. SSC-related lung disease is highly associated with early mortality in patients with systemic sclerosis. A U.S. national survey showed that the mortality rate from pulmonary fibrosis in patients with systemic sclerosis increased more than fivefold over a 20-year period, from 6% to 33%, making interstitial lung disease the most frequent cause of systemic sclerosis-related death. EULAR survey results also revealed that interstitial lung disease contributes to the mortality rate of more than 35% of patients with systemic sclerosis. (Steen VD, Medsger TA. Changes in causes of death in systemic sclerosis, 1972-2002. Ann Rheum Dis. 2007;66:940-4)

[0006] Risk factors for the development or progression of interstitial lung disease in patients with systemic sclerosis include the presence of diffuse cutaneous systemic sclerosis (dcSSc), African American race, older age at onset, shorter disease duration, and the absence of anti-Scl-70 / anti-topoisomerase I and anti-centromere antibodies. ILD develops when the conditions for its onset are met for various reasons that remain to be elucidated. However, none of these risk factors are absolute, and the development of systemic sclerosis does not automatically lead to systemic sclerosis-related lung disease. Furthermore, because not all patients exhibit respiratory symptoms, physicians and researchers are using various methods to diagnose the disease early and develop treatments (Respiratory Research Volume 20, Article Number: 13 (2019)).

[0007] In particular, recent studies using clinical samples have shown that systemic sclerosis-associated lung disease, derived from systemic sclerosis patients, shows significant differences in various cytokines and biomarkers such as IGFBP-1, MMP-9, and H3.1 in the blood, suggesting different disease characteristics in patients. These studies are currently being continuously conducted to utilize biomarkers for diagnosing systemic sclerosis-associated lung disease in systemic sclerosis cohorts, and this data could serve as the basis for considering the two diseases as separate, though highly related, classifications. Furthermore, the fact that MMP-7 and MMP-9 are specifically elevated in the blood of patients with systemic sclerosis-associated lung disease compared to patients with systemic sclerosis provides evidence supporting the reasons for the onset of pulmonary fibrosis and the rapid and high mortality rate of patients with systemic sclerosis-associated lung disease, which accounts for up to one-third of patients. This finding is also interpreted as evidence for the clinical and pathological distinction between systemic sclerosis-associated lung disease and systemic sclerosis ((i) Clin Epigenetics. 2020 Aug 17; 12 (1): 124, (ii) Sarcoidosis Vasc Diffuse Lung Dis. 2015 Sep 14; 32 (3): 228-36, (iii) European Respiratory Journal 2021 58: 2101560).

[0008] To date, there is no cure for systemic sclerosis that can halt the progression of the disease, but some medications can alleviate specific symptoms and reduce organ damage. Nonsteroidal anti-inflammatory drugs (NSAIDs) help relieve joint pain, and calcium channel blockers can alleviate the symptoms of Raynaud's phenomenon, but they can also cause gastrointestinal problems. Corticosteroids can alleviate myositis symptoms, immunosuppressants can alleviate pulmonary inflammation, and hypertension medications can alleviate acute kidney injury and elevated blood pressure, but these medications only partially alleviate symptoms, and no medications offer comprehensive symptom relief.

[0009] For systemic sclerosis-associated lung disease, Ofev® (nintedanib) and Actemra® (tocilizumab) have been approved as therapeutic agents. Nintedanib is known to improve lung function in patients with idiopathic pulmonary fibrosis. However, nintedanib is only effective in improving pulmonary fibrosis and has failed to demonstrate therapeutic efficacy for skin sclerosis symptoms. Therefore, it is only prescribed as an adjunctive treatment for improving lung function in patients with end-stage systemic sclerosis-associated lung disease who already have progressive interstitial lung disease. Tocilizumab is an immunosuppressant for treating rheumatoid arthritis and is known to alleviate the symptoms of rheumatic diseases. However, tocilizumab has not been proven to alleviate / improve fibrosis and is used as a preventative adjunctive treatment in patients with early-stage systemic sclerosis-associated lung disease who have not yet experienced severe lung function impairment. In other words, neither of the approved drugs can provide fundamental therapeutic effects for patients with systemic sclerosis or systemic sclerosis-associated lung disease, so they are not prescribed as first-line treatment, and patients must still rely on immunosuppressants such as cyclophosphamide and mycophenolate, which have severe side effects.

[0010] Meanwhile, prolyl-tRNA synthetase (PRS) is a member of the aminoacyl-tRNA synthetase (ARS) family of enzymes that activates amino acids for protein synthesis. After forming aminoacyl adenylate (AA-AMP), ARS performs the translational function of transferring the activated amino acid to the 3-terminal end of the corresponding tRNA. Because ARS plays a crucial role in protein synthesis, ARS inhibition suppresses the growth and development of all cells. Therefore, ARS is recognized as a promising target for antibiotics and therapeutic agents for diseases that require the suppression of cellular overexpression (Nature, 2013, 494:121-125).

[0011] PRS exists and functions in the form of EPRS (Glutamyl-Prolyl-tRNA Synthetase) in the multisynthetase complex (MSC). In particular, EPRS functions as a translational silencer that suppresses the production of VEGF A (vascular endothelial growth factor A), a key factor in angiogenesis, among various MSCs. It has also been reported to be closely associated with various solid cancers (Nat. Rev. Cancer, 2011, 11, 708-718).

[0012] Therefore, the present inventors have conducted extensive research into methods for preventing or treating systemic sclerosis, and as a result have found that the specific PRS inhibitors described below are useful for preventing or treating systemic sclerosis, thereby completing the present invention. Summary of the Invention [Problem to be solved by the invention]

[0013] The present invention provides a pharmaceutical composition that can be usefully used for the prevention or treatment of systemic sclerosis. [Means for solving the problem]

[0014] In order to solve the above problems, the present invention provides the following pharmaceutical composition for preventing or treating systemic sclerosis: A pharmaceutical composition for preventing or treating systemic sclerosis, comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof: [ka]

[0015] The compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof is a compound described in Korean Patent Registration No. 10-2084772, specifically, the substance described in Example 40 of the specification.

[0016] The pharmaceutical composition according to the present invention is useful for preventing or treating systemic sclerosis, particularly systemic sclerosis-associated interstitial lung disease (SSc-ILD). In particular, the pharmaceutical composition according to the present invention has the dual effects of alleviating skin stiffness and improving lung function in systemic sclerosis-associated lung disease. The term "improvement of lung function" used in the present invention refers to an improvement in the ability to supply blood to the body, which is essentially a basic function of the lungs. This is not an effect that necessarily accompanies improvement of pulmonary fibrosis, and can be confirmed by body oxygen saturation, separately from pulmonary fibrosis indicators.

[0017] As used herein, the term "prevention" refers to any action that inhibits or delays the occurrence, spread, and recurrence of the disease by administering the composition of the present invention, and "treatment" refers to any action that improves or favorably alters the symptoms of the disease by administering the composition of the present invention.

[0018] Meanwhile, the compound represented by Chemical Formula 1 can be used in the form of a pharmaceutically acceptable salt, and useful salts include acid addition salts formed with a pharmaceutically acceptable free acid. The free acid can be an inorganic or organic acid. Examples of inorganic acids that can be used include hydrochloric acid, bromic acid, sulfuric acid, and phosphoric acid, while examples of organic acids that can be used include citric acid, acetic acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, succinic acid, 4-toluenesulfonic acid, glutamic acid, and aspartic acid. Preferably, the pharmaceutically acceptable salt of the compound represented by Chemical Formula 1 is a hydrochloride salt.

[0019] Furthermore, the compound represented by the formula (1) above can be prepared in a crystalline or non-crystalline form, and when prepared in a crystalline form, it can be optionally hydrated or solvated. The present invention includes not only stoichiometric hydrates of the compound represented by the formula (1) above, but also compounds containing various amounts of water. The solvates of the compound represented by the formula (1) above include both stoichiometric and non-stoichiometric solvates.

[0020] The pharmaceutical compositions of the present invention can be formulated into oral or parenteral dosage forms according to standard pharmaceutical practice. These dosage forms may contain, in addition to the active ingredient, additives such as pharmaceutically acceptable carriers, adjuvants, or diluents.

[0021] Suitable carriers include, but are not limited to, physiological saline, polyethylene glycol, ethanol, vegetable oil, and isopropyl myristate, and diluents include, but are not limited to, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and / or glycine. The compound of the present invention can also be dissolved in oil, propylene glycol, or other solvents commonly used in the preparation of injection solutions. For topical application, the compound of the present invention can also be formulated into ointments or creams.

[0022] The pharmaceutical dosage forms of the compounds of the present invention may be in the form of their pharmaceutically acceptable salts or solvates, and may be used alone or in combination with other pharmaceutically active compounds, or in suitable combinations. For example, the pharmaceutical composition of the present invention may additionally contain other active ingredients used in the prevention or treatment of systemic sclerosis. Examples of such other active ingredients include Ofev® (nintedanib) or Actemra® (tocilizumab). When the pharmaceutical composition of the present invention additionally contains other active ingredients, the weight ratio of the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof to the other active ingredients is preferably 1:0.1 to 1:10.

[0023] The compounds of the present invention can be formulated into injections by dissolving, suspending, or emulsifying them in aqueous solvents such as common saline, 5% dextrose, or non-aqueous solvents such as synthetic fatty acid glycerides, higher fatty acid esters, or propylene glycol. The dosage forms of the present invention can contain conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers, and preservatives.

[0024] The pharmaceutical composition according to the present invention can be administered orally or parenterally. Depending on the administration method, the composition according to the present invention can contain 0.001 to 99% by weight, preferably 0.01 to 60% by weight, of the compound represented by Chemical Formula 1 or a pharmaceutically acceptable salt thereof.

[0025] The pharmaceutical composition of the present invention can be administered to mammals, including mice, rats, livestock, and humans, by a variety of routes, including oral, rectal, intravenous, intramuscular, subcutaneous, intrauterine, intradural, or intracerebroventricular injection, although any route of administration is contemplated. [Effects of the Invention]

[0026] As described above, the pharmaceutical composition according to the present invention can be useful for preventing or treating systemic sclerosis. [Brief explanation of the drawings]

[0027] [Figure 1] FIG. 1 shows the results of confirming the thickness of 3D skin organoids by H&E immunochemical staining in Experimental Example 1 of the present invention. [Figure 2] FIG. 1 shows the results of immunochemical staining of the skin thickness of a systemic dermal sclerosis skin organoid transplant mouse model in Experimental Example 2 of the present invention. [Figure 3] FIG. 1 shows the results of immunochemical staining to confirm the presence of fibrotic factors Collagen I, Collagen III, and αSMA in the skin tissue of a systemic dermal sclerosis skin organoid transplant mouse model in Experimental Example 2 of the present invention. [Figure 4] FIG. 1 shows the experimental schedule for mice in Experimental Example 3 of the present invention. [Figure 5] FIG. 1 shows the results of changes in mouse body weight in Experimental Example 3-1 of the present invention. [Figure 6]FIG. 1 shows the results of changes in lung weight in mice in Experimental Example 3-2 of the present invention. [Figure 7] FIG. 1 shows the results of lung function evaluation of mice in Experimental Example 3-3 of the present invention. [Figure 8] FIG. 10 is a diagram showing the results of evaluation of mouse skin thickness in Experimental Example 3-4 of the present invention. [Figure 9] FIG. 1 shows the results of measuring collagen content in mice in Experimental Example 3-5 of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0028] The present invention will be described in more detail with reference to the following examples, but the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0029] Example The following compounds were prepared in the same manner as in Example 40 of Korean Patent Registration No. 10-2084772, and are hereinafter referred to as "active ingredients" or "Examples." [ka] 1 H NMR (500 MHz, MeOD): δ 9.67 (s, 1H). 8.02 (d, 1H), 7.82 (d, 1H), 4.62 (m, 2H), 3.60 (m, 1H), 3.28 (m, 1H), 2.99 (m, 2H), 2.25 (m, 2H), 2.08 (m, 2H), 1.99 (m, 1H), 1.78 (m, 2H), 1.54 (m, 1H)

[0030] Experimental example 1: Confirmation of skin hardening alleviation effect in a 3D skin organoid model Using iPSC-derived fibroblasts from a patient with systemic dermatosclerosis, 3D skin organoids were created using the method described in a literature review (Lancet. 2009 Nov 21; 374 (9703): 1745-53). Treatment with 1 μM of the active ingredient in DMSO solution reduced the thickness of the 3D skin organoids, as confirmed by H&E immunochemical staining (Figure 1). The control group was treated with DMSO only.

[0031] Experimental example 2: Confirmation of the skin hardening alleviation effect in a 3D skin organoid transplant mouse model A mouse model of systemic dermatosclerosis (SDS) was constructed by transplanting a previously constructed 3D skin organoid derived from iPSCs from a patient with SDS into SCID mice. Mouse skin tissue was cut into 1x2cm pieces and sutured to the mouse skin. The skin was transplanted onto the back of the mouse using the clinically used tie-over dressing method. Gauze was placed over the sutured area, which was then tied off with sutures and dressed with a band.

[0032] The active ingredient was administered subcutaneously at 3 mg / kg or 10 mg / kg daily for two weeks to a mouse model of systemic dermal sclerosis (SDS) with skin organoid transplants. Normal saline was administered to the normal skin tissue of the negative control group (NC), and saline was administered to the skin tissue of the positive control group (SSc) with SDS transplants. Skin thickness, the primary endpoint of SDS, was measured using immunochemical staining, and the results are shown in Figure 2. As shown in Figure 2, the active ingredient significantly reduced skin thickness (***p<0.001).

[0033] To confirm whether the active ingredient reduces fibrotic factors in systemic dermatosclerosis skin tissue, immunochemical staining was performed, and the results are shown in Figure 3. As shown in Figure 3, it was confirmed that the active ingredient significantly reduces the fibrotic factors Collagen I, Collagen III, and αSMA (*p<0.05, ***p<0.001).

[0034] Experimental Example 3: Confirmation of therapeutic effects in a mouse model of systemic sclerosis-associated lung disease (SSc-ILD) Seven-week-old C57BL / 6 mice were obtained and allowed to acclimate indoors for at least 5 days. An osmotic pump-based systemic sclerosis-associated lung disease model was designed. Without the osmotic pump, BLM would have to be subcutaneously injected into the back of the test animals daily for 4 weeks, which would have caused severe pain at the injection site.

[0035] As shown in Figure 4, the drug administration rate using the osmotic pump was 0.5 μl / hr (7 days), and the size of the osmotic pump was 1.5 cm. The incision was sutured, the osmotic pump was inserted, and bleomycin was administered for one week. The osmotic pump was then removed on the 10th day. Systemic sclerosis-related lung disease was confirmed to be induced in the BLM-administered mouse model for one week. The specific experimental groups are shown in Table 1 below.

[0036] [Table 1]

[0037] Experimental Example 3-1: Body weight (%) Based on the results of 28 days of active ingredient administration (Table 2 and Figure 5), significant changes in body weight were observed from 3 mg / kg of active ingredient onwards, and at 30 mg / kg, body weight increased significantly compared to the vehicle (p<0.01).

[0038] [Table 2]

[0039] Experimental Example 3-2: Lung weight Changes in lung weight showed a significant trend only at 30 mg / kg, but after adjustment for body weight, significance emerged from 10 mg / kg and was confirmed to be higher at 30 mg / kg with p<0.01 (Table 3 and Figure 6).

[0040] [Table 3]

[0041] Experimental Example 3-3: Lung Function Evaluation (SpO2 (%)) This pulmonary function assessment is an experiment to demonstrate the therapeutic effect of this active substance in patients with systemic sclerosis-related lung disease, and unlike the general effect of improving pulmonary fibrosis, it is the most direct and important evaluation index that has the greatest impact on the symptoms and quality of life of patients with systemic sclerosis-related lung disease. This experiment can most directly demonstrate the effect of improving lung function in an animal model of systemic sclerosis-related lung disease by measuring the internal oxygen concentration.

[0042] On day 28, SpO2 was measured using a veterinary pulse oximeter (Berry) through the abdominal side of the mouse. The results are shown in Figure 7.

[0043] As shown in Figure 7, significant results were observed from 10 mg / kg of the active ingredient, and at 30 mg / kg, oxygen saturation was higher than the vehicle (p<0.001). Furthermore, it was confirmed that the active ingredient 30 mg / kg group showed an oxygen saturation recovery rate equivalent to that of the comparison group, nintedanib 60 mg / kg.

[0044] Experimental Example 3-4: Skin thickness evaluation On day 28, skin tissue was extracted from the mice and subjected to H&E and Mansson's Trichrome (MT stain). The stained photographs were then used to measure skin thickness at 10 random locations (at a minimum interval of 200-300 μm) using ImageJ. Measurements were made as uniformly as possible across the entire photograph, and areas with damaged stained tissue were excluded. The results are shown in Figure 8 and Table 4.

[0045] As shown in Figure 8, in the BLM group, collagen was fully induced not only in the dermoid skin but also in areas where adipose tissue should be present. Significant results were observed from 10 mg / kg of the active ingredient, and highly significant results were observed at 30 mg / kg (P<0.0001). Furthermore, overall, the level of MT staining was low when the active ingredient was administered, indicating a decrease in collagen density within the skin. While skin thickness did not decrease significantly at 3 mg / kg of the active ingredient, a tendency toward a decrease in collagen density was confirmed. In particular, 10 mg / kg of the active ingredient demonstrated a superior improvement in skin thickness compared to nintedanib at 60 mg / kg.

[0046] [Table 4]

[0047] Experimental Example 3-5: Measurement of collagen content in the body We attempted to indirectly measure collagen content by measuring the amount of hydroxyproline in the mouse body. Analysis was performed using an insoluble collagen assay (Biocolor, S2000). Specifically, the results were as follows:

[0048] First, 10 mg of frozen skin tissue was crushed with 100 μl of Fragmentation Reagent. 100 μl of 37% HCl was added, and the mixture was incubated at 65°C for 3 hours. The contents of the tube were shaken every 30 minutes to aid in tissue disintegration. The mixture was centrifuged at 12,000 rpm for 10 minutes. After centrifugation, the mixture was transferred to a 1.5 ml tube. 50 μl of the sample between 10 and 100 μl was taken, and distilled water was added to bring the total volume to 100 μl to prepare the sample.

[0049] 1 ml of dye was added to 100 μl of the prepared sample and mixed for 30 minutes. After centrifugation at 12,000 rpm for 10 minutes, the sample was transferred to a new tube. The dye was removed with 750 μl of ice-cold Acid-Salt Wash Reagent. After centrifugation at 12,000 rpm for 10 minutes, the sample was transferred to a new tube. The tube was then mixed with the collagen-binding dye using a vortex mixer. The collagen-binding dye dissolved within 10 minutes and the measurement was complete (measurements should be completed within 2-3 hours). The tube was kept closed until the absorbance was ready to be measured.

[0050] 200 μl of each sample was transferred to individual wells of a 96-microwell plate, and the absorbance was measured at 560 nm. The hydroxyproline values ​​were compared with the normal group and the results were expressed as a ratio. The results are shown in Table 5 and Figure 9.

[0051] The BLM group showed a more than two-fold increase in skin hydroxyproline compared to the saline group, confirming good modeling. Significant results were observed from 10 mg / kg of the active ingredient, and at 30 mg / kg, highly significant results were observed (P<0.001).

[0052] [Table 5]

[0053] As can be seen from the above experimental results, the active ingredient of the present invention exhibited two completely different effects: a therapeutic effect for systemic sclerosis and an improvement in the inhibition of pulmonary function in systemic sclerosis-related lung diseases. Such dual effects cannot be predicted from the therapeutic effect on either one, and are considered to be a characteristic of the active ingredient of the present invention.

Claims

1. A pharmaceutical composition for treating systemic sclerosis associated interstitial lung disease (SSc-ILD), comprising a compound represented by the following Chemical Formula 1 or a pharmaceutically acceptable salt thereof: 【Chemistry 1】

2. The pharmaceutical composition has the dual effect of alleviating skin hardening and improving lung function in systemic sclerosis-related interstitial lung disease. The pharmaceutical composition of claim 1.

3. The pharmaceutically acceptable salt is a hydrochloride salt. The pharmaceutical composition of claim 1.

4. The pharmaceutical composition additionally contains another active ingredient used in the prevention or treatment of systemic sclerosis. The pharmaceutical composition of claim 1.

5. The other active ingredient used in the prevention or treatment of systemic sclerosis is nintedanib or tocilizumab. The pharmaceutical composition of claim 4.

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