TGF-beta receptor inhibitors for use in the therapeutic treatment of a pulmonary fibrotic disease

Abstract

The present invention relates to a pharmaceutical composition comprising a TGF-beta receptor inhibitor selected from the group consisting of Galunisertib (LY2157299), Vactosertib (EW-7197), Itacnosertib (TP-0184), and Repsox (E-616452), and at least one pharmaceutically acceptable carrier, excipient and / or diluent, for use in the treatment of a fibrotic lung disease, in particular idiopathic pulmonary fibrosis (IPF). Preferably, the at least one excipient of the pharmaceutical composition for use according to the invention comprises triterpenoid saponin molecules, more preferably asiaticoside and / or glycyrrhizin molecules.

Description

[0001] TGF-beta receptor inhibitors for use in the therapeutic treatment of a fibrotic lung disease

[0002] The present invention falls within the field of therapeutic treatments of fibrotic lung diseases, in particular idiopathic pulmonary fibrosis (IPF).

[0003] Interstitial lung diseases are a heterogeneous group of diseases (more than 300) characterized by cell proliferation, interstitial inflammation, fibrosis, or a combination of such events within the alveolar wall. Interstitial fibrosis represents the predominant phenotype in most cases. Most patients with interstitial fibrosis are diagnosed with chronic (hypersensitivity) pneumonia, pulmonary sarcoidosis, underlying autoimmune disease or, in the absence of an identified cause, idiopathic interstitial pneumonia (Lederer D et al, “Idiophatic pulmonary fibrosis” The new england journal of medicine, 2018, 378:1811-23).

[0004] Several mechanisms underlie the pathogenesis of the development of interstitial fibrosis. Typically, the process is triggered by repeated inflammatory damage to the epithelium and vascular system, which not only causes cell injury but also affects the associated repair system. In fact, fibroblasts migrate from the pulmonary epithelium and peripheral circulation to the site of the injury, which leads to their uncontrolled proliferation, differentiation and activation, causing the loss of the normal architecture of the lung parenchyma.

[0005] In addition, pro-fibrotic mediators are released from macrophages and lymphocytes, resulting in increased alveolar tissue stiffness. Subsequent activations of the fibroblasts trigger a continuous cycle of progressive pulmonary fibrosis. Pro-fibrotic mediators such as platelet-derived growth factor (PDGF), tumor necrosis factor-a (TNF-a), transforming growth factor-P (TGF-P) and matrix metalloproteinases (MMPs) contribute to the pathogenesis process (Bagnato G et al. “Cellular interactions in the pathogenesis of interstitial lung diseases”. Eur Respir Rev. 2015; 24:102-14; Shumar JN et al. “Antifibrotic Therapies and Progressive Fibrosing Interstitial Lung Disease (PF-ILD): Building on INBUILD”. J Clin Med. 2021; 10:2285).

[0006] The most common idiopathic interstitial pneumonia is idiopathic pulmonary fibrosis (IPF), an irreversible, chronic fibrotic interstitial lung disease of unknown cause characterized by progressive pulmonary function impairment. The main symptoms include the gradual onset of shortness of breath, progressive dyspnea, and dry, non-productive cough. This disease affects about 3 million people worldwide, with an incidence that increases with age (Raghu G et al. “Diagnosis of idiopathic pulmonary fibrosis. An Official ATS / ERS / JRS / ALAT clinical practice guideline”, Am J Respir Crit Care Med, 2018 (5), 44-68).

[0007] The main drugs of choice for the treatment of interstitial lung diseases are pirfenidone and nintedanib.

[0008] Pirfenidone (5-methyl-l-phenyl-2-(lH)-pyridone) is a molecule with antifibrotic, antiinflammatory and antioxidant activity. It works by reducing collagen production, slowing down the fibrotic process through inhibition of transforming growth factor-beta (TGF-beta) and decreasing the decline in forced vital capacity (FVC). As is known, the FVC parameter indicates the total volume of air expelled with a forced expiration, starting from a maximal inspiration. As regards the dosage, pirfenidone is generally administered orally at a dosage of 2,403 mg / day.

[0009] After oral administration, pirfenidone is absorbed by the gastrointestinal tract and approximately 60% of it binds to plasma proteins, particularly albumin (Taniyama M et al., “Pharmacokinetics of an antifibrotic agent, pirfenidone, in haemodialysis patients”, Eur. J. Clin. Pharmacol., 1997,52, 77-8). Hepatically metabolized by CYP1A2 to the inactive metabolite 5-carboxypirfenidone (approximately 50%), the only metabolite present in the plasma in significant quantities, it is eliminated in urine as a carboxylic metabolite (within 24 hours of intake).

[0010] However, oral administration of pirfenidone is accompanied by significant side effects, especially at the gastrointestinal level, with loss of appetite, nausea, vomiting and gastroesophageal reflux. Other side effects reported include lung infections, skin rash and lethargy. In some cases, an increase in aminotransferase levels has also been observed, requiring dosage modification or discontinuation of therapy.

[0011] High dosages of systemic therapy are necessary to compensate for the first-pass effect, systemic distribution, low potency and thus to achieve a therapeutic concentration of pirfenidone at the lung level (Kaminskas L et al. “Aerosol Pirfenidone Pharmacokinetics after Inhaled Delivery in Sheep: a Viable Approach to Treating Idiopathic Pulmonary Fibrosis”. Pharm Res (2020) 37:3).

[0012] Nintedanib (Methyl (3Z)-3-(((4-(N-methyl-2-(4-methylpiperazin- 1-yl) acetamido)phenyl)amino)(phenyl)methylidene)-2-oxo-2,3-dihydro-lH-indole-6-carboxylate) is an intracellular inhibitor of tyrosine kinase receptors, platelet-derived growth factor receptors (PDGFRs), fibroblast growth factor receptors (FGFRs), and vascular endothelial growth factor receptors (VEGFRs). It is effective in reducing FVC decline and acute exacerbations in patients with IPF. The recommended dose is 150 mg twice a day by oral administration.

[0013] Taking nintedanib is also associated with the appearance of side effects, especially at the gastrointestinal level, such as diarrhea, nausea, decreased appetite, abdominal pain, as well as an increase in liver enzyme levels during the first year of treatment (Mazzei M et al. “Nintedanib in the treatment of idiophatic pulmonary fibrosys” Therapeutic advances in repsiratory disease. 2015, 9 121:129).

[0014] Oral administration of nintedanib results in low bioavailability (4.7%) mainly due to the first-pass effect and P-glycoprotein-mediated cellular efflux (Kala S et al, “Bioavailability enhancement of vitamin E TPGS liposomes of nintedanib esylate: formulation optimization, cytotoxicity and pharmacokinetic studies”, Drug Deliv Transl Res, 2022, 12(11):2856-2864). However, increasing the dosage is not a plausible option due to side effects.

[0015] Nintedanib is metabolized through hydrolytic cleavage by esterases causing the formation of carboxylic acid, which is subsequently glucuronidated by glucuronyltransferase enzymes. Only to a lesser extent is nintedanib metabolized by cytochrome pathways, with CYP3A4 being the predominant enzyme involved. However, the CYP-dependent metabolite has not been detected in the plasma during absorption, distribution, metabolism. The main clearance route is by fecal / biliary excretion (approximately 90% of the dose) mainly in the form of free acid. Further new therapies for IPF are currently under development. The authors of the scientific paper Glass et al, “Idiopathic pulmonary fibrosis: Current and future treatment”, Clin Respir J. 2022; 16(2): 84-96, describe recent therapeutic approaches based on the use of pentatraxin, autotaxin inhibitors, the anti-connective tissue growth factor monoclonal antibody pamrevlumab, galectin-3 inhibitors, human R antigen inhibitors, as well as the use of cell therapies.

[0016] In recent years, inhibition of the TGF-beta signalling pathway has assumed an important role in the clinical setting as a pharmacological target. This development, however, is accompanied by considerable difficulties as TGF-beta is a pleiotropic cytokine which also plays important roles under physiological conditions, particularly in cardiac development. Studies by Anderton MJ et al., “Induction of heart valve lesions by small-molecule ALK5 inhibitors”, Toxicol Pathol, 2011 39(6):916-24, have shown, for example, that small TGF-beta receptor I kinase (ALK5) inhibitor molecules also cause cardiac toxicity in adult animals, suggesting an important homeostatic role of ALK5 in maintaining the integrity of heart valves.

[0017] Effects related to cardiac toxicity have also been demonstrated for anti-TGF-beta monoclonal antibodies (Mitra M et al., “A Potent Pan-TGFP Neutralizing Monoclonal Antibody Elicits Cardiovascular Toxicity in Mice and Cynomolgus Monkeys”, Toxicological Sciences, 2020, 175, 24-34).

[0018] In this context, therefore, there is a dramatic need for therapeutic strategies that are effective in treating fibrotic lung diseases, in particular idiopathic pulmonary fibrosis (IPF), by counteracting the onset and progression thereof, and that are suitable for achieving a lasting clinical response while minimizing possible adverse effects. There is also a need to provide therapeutic strategies that allow for targeted delivery of the therapeutic agent to the target lung tissues, thereby overcoming the limitations of the formulation strategies of the prior art as previously illustrated.

[0019] This need has now been met by the present inventors, who have found that TGF-beta receptor inhibitors selected from galunisertib (LY2157299), vactosertib (EW-7197), itacnosertib (TP-0184), and repsox, are able to stop the process of differentiation of fibroblasts into myofibroblasts, which is characteristic of the fibrotic process leading to lung dysfunction, as well as to exert an inhibitory activity on the expression of proteins that play a primary role in the aforementioned process, such as a-SMA actin, fibronectin, and vimentin.

[0020] Therefore, one object of the present invention is a pharmaceutical composition comprising a TGF-beta receptor inhibitor, preferably a TGF-beta receptor type 1 inhibitor, selected from the group consisting of galunisertib (LY2157299), vactosertib (EW-7197), itacnosertib (TP-0184), and repsox, and at least one pharmaceutically acceptable carrier, excipient and / or diluent, wherein the at least one pharmaceutically acceptable excipient comprises one or more triterpenoid saponin molecules, for use in the therapeutic treatment of a fibrotic lung disease.

[0021] Preferably, the fibrotic lung disease according to the invention is selected from the group consisting of idiopathic pulmonary fibrosis (IPF), non-specific interstitial pneumonia (NSIP), chronic hypersensitivity pneumonitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), pulmonary sarcoidosis, pneumoconiosis, cryptogenic organizing pneumonia, interstitial lung disease associated with autoimmune diseases, and any combination thereof. More preferably, said lung disease is idiopathic pulmonary fibrosis (IPF).

[0022] Further features and advantages of the invention are identified in the appended claims and illustrated in detail in the following description.

[0023] The appended independent and dependent claims form an integral part of the present specification.

[0024] Within the scope of the present description, the term “TGF-beta receptor inhibitor” refers to a compound capable of stopping, blocking, inhibiting, partially or completely neutralizing the transforming growth factor beta (TGF-beta) receptor and the signalling pathway downstream of said receptor, which is known to be involved in the cellular processes of proliferation, differentiation, and apoptosis.

[0025] According to the invention, the TGF-beta receptor inhibitor is selected from the following compounds:

[0026] Galunisertib, CAS number 700874-72-2; synonyms: LY2157299, 4-(2-(6-methylpyridin-2-yl)-5,6-dihydro-4H-pyrrole-[l,2-b]pyrazol-3-yl)quinoline-6-carboxyamide);

[0027] Vactosertib, CAS number 1352608-82-2; synonyms: EW-7197, N-(2-fluorophenyl)-5-(6-methyl-2-pyridinyl)-4-[l,2,4]triazole[l,5-a]pyridin-6-yl-lH-imidazole-2-methanamine; Itacnosertib, CAS number 1628870-27-8; synonyms: TP-0184, 2-A-[3-methoxy-4-(4-methylpiperazin-l-yl)phenyl]-4-A-(2-pyridin-2-ylpyridin-3-yl)pyrimidin-2,4-diamine; and Repsox, CAS number 446859-33-2; synonyms: E-616452, 2-[3-(6-Methyl-2-pyridinyl)-lH-pyrazol-4-yl] - 1 ,5-naphthyridine.

[0028] In a preferred embodiment according to the invention, the TGF-beta receptor inhibitor is galunisertib.

[0029] Among the above-mentioned TGF-beta signalling pathway inhibitors, galunisertib has been the subject of clinical trials by oral administration for its antitumor activity, but the development of this drug was discontinued during Clinical Phase II.

[0030] According to the invention, it is understood that any possible combination of the TGF-beta receptor inhibitors in the pharmaceutical composition as defined above, is covered by the present invention.

[0031] A preferred pharmaceutical composition for use according to the invention comprises galunisertib.

[0032] Another preferred pharmaceutical composition for use according to the invention comprises galunisertib and vactosertib.

[0033] A further preferred pharmaceutical composition for use according to the invention comprises galunisertib and repsox.

[0034] Yet another preferred pharmaceutical composition for use according to the invention comprises the TGF-beta receptor inhibitors galunisertib, vactosertib and repsox.

[0035] Preferably, the pharmaceutical composition for use according to the invention is in a pharmaceutical form suitable for administration via inhalation.

[0036] This advantageously overcomes important limitations associated with other routes of administration of the above-mentioned TGF-beta inhibitors, in particular the limitations associated with oral administration. Pre-clinical toxicity studies conducted on galunisertib have, in fact, revealed that prolonged oral intake of this drug is accompanied by a high risk of cardiac toxicity. As a result of this observation, a specific administration regimen was introduced into the clinical protocol which involved taking galunisertib for 14 consecutive days, followed by a discontinuation of therapy for the next 14 days (Herbertz S et al., “Clinical development of galunisertib (LY2157299 monohydrate), a small molecule inhibitor of transforming growth factor-beta signaling pathway”, Drug Des Devel Ther.

[0037] 2015; 9: 4479-4499).

[0038] In addition, the inhalation administration of the pharmaceutical composition for use according to the invention, which is therefore not affected by the known first-pass effect and / or gastric degradation, advantageously allows for a reduction in the TGF-beta receptor inhibitor dosage used, thereby reducing the risk of dangerous side effects, such as the aforementioned cardiotoxicity. A further advantage of this administration method is the achievement of a high concentration of the active ingredient directly in the lung tissues, despite using lower drug dosages than those typically used for oral administration.

[0039] Pharmaceutical formulations suitable for inhalation administration may be in powder or liquid form.

[0040] Inhalation administration of the pharmaceutical composition for use according to the invention may be carried out by means of suitable devices such as, for example, nebulizers, dry powder inhalers (DPIs), and pressurized metered dose inhalers (pMDIs), or any other device for inhalation use which is known per se to those of ordinary skill in the art.

[0041] In the case of administration via a pMDI device, a gaseous propellant is generally used, typically hydrofluoroalkanes (HFAs). Other main components of a pMDI are an aluminum can, containing a plastic shell with a mouthpiece, a dispensing valve, and an activator. The generation of aerosol by the pMDI devices is due to the presence of the propellant, without requiring particularly high inspiratory flows from the patient. By pressing the can, the drugpropellant mixture is released into the activator, then expansion and vaporization convert the drug into an aerosol.

[0042] Alternatively, the pharmaceutical composition for use according to the invention may be administered by inhalation using a nebulizer or a dry powder inhaler (DPI).

[0043] DPI devices do not have a propellant to generate the aerosol, and the required energy is produced directly by the patient when he / she inhales into the inhaler. They can be configured as single-dose (e.g. pre-dosed capsules) or multi-dose.

[0044] The selection of the most suitable device for use within the scope of the present invention for the administration of the pharmaceutical composition by inhalation falls well within the skills of those of ordinary skill in the art.

[0045] In one embodiment, the pharmaceutical form of the pharmaceutical composition for use according to the invention is an inhalable dry powder.

[0046] Preferably, the inhalable dry powder according to the invention consists of particles having a mass median aerodynamic diameter (MMAD) distribution ranging from 1pm to 10pm, preferably from 1pm to 5pm.

[0047] The term “mass median aerodynamic diameter (MMAD)”, as used herein, refers to the statistical aerodynamic diameter value above and below which 50% of the particles are contained. Advantageously, the aforementioned particle size distribution in the inhalable dry powder allows the deposition of the particles in the pulmonary alveoli.

[0048] In one embodiment, the fine particle fraction (FPF) of the inhalable dry powder according to the invention is equal to or greater than 40%, or equal to or greater than 50%, or equal to or greater than 60%, or equal to or greater than 70%, or equal to or greater than 80%, or equal to or greater than 90%, or equal to or greater than 95%.

[0049] The term “fine particle fraction (FPF)”, as used herein, refers to the fraction of aerosol particles (given as a percentage) having a diameter less than 5 pm, as measured by means of a cascade impactor.

[0050] Methods suitable for producing an inhalable dry powder are known and described in the state of the art, therefore the selection and use thereof are well within the skills of those of ordinary skill in the art. The spray drying procedure is mentioned as a non-limiting example.

[0051] According to one embodiment, the pharmaceutical composition for use according to the invention further comprises a therapeutic agent selected from the group consisting of corticosteroids, antifibrotic agents, phosphodiesterase inhibitors, immunosuppressive agents, and any combination thereof.

[0052] Antifibrotic agents suitable for use according to the invention include, but are not limited to, pirfenidone and nintedanib.

[0053] Corticosteroids suitable for use according to the invention include, for example, beclomethasone, budesonide, dexamethasone, and fluticasone.

[0054] Phosphodiesterase inhibitors suitable for use according to the invention include, for example, roflumilast.

[0055] Immunosuppressive agents suitable for use according to the invention include, for example, mycophenolic acid, cyclophosphamide, and azathioprine.

[0056] According to a preferred embodiment, in the pharmaceutical composition for use according to the invention, the at least one pharmaceutically acceptable excipient comprises saponin molecules, preferably triterpenoid saponin molecules.

[0057] As is known, saponins are naturally occurring glycosides consisting of a sugar portion and an aglycone which, depending on the chemical structure, can be classified as steroidal, triterpenic, or glycoalkaloid. More specifically, triterpenoid saponins consist of a C30 pentacyclic terpene to which one or more sugars are attached.

[0058] Preferably, the triterpenoid saponin molecules according to the invention are selected from the group consisting of asiaticoside, astragaloside IV, ginsenoside Rgl, glycyrrhizin, and any combination thereof.

[0059] Asiaticoside and / or glycyrrhizin are particularly preferred triterpenoid saponins.

[0060] The asiaticoside saponin has the following chemical structure:

[0061]

[0062] This saponin (C48H78019, molecular weight 959.12 g / mol) is extracted from the herb Centella asiatica and is characterized by poor water solubility (0.798 mg / ml, Drug Bank source), reduced plasma half-life, and limited ability to cross physiological barriers. Despite some attempts to use saponins as solubilizing agents in pharmaceutical formulations (Heng Yang F et al. “Bioavailability Enhancement of Paclitaxel via a Novel Oral Drug Delivery System: Paclitaxel-Loaded Glycyrrhizic Acid Micelles”, Molecules, 2015, 20 (3), 4337-4356; Wang Y et al. “Formulation and evaluation of novel glycyrrhizic acid micelles for transdermal delivery of podophyllo toxin”, Drug Deliv, 2016; 23(5): 1623-1635), this use still remains of limited applicability. In fact, saponins exhibit a dose-dependent haemolytic effect which makes their intravenous administration difficult. Furthermore, the solubilizing activity of saponins is strictly dependent on the type of saponin as well as on the type of molecule the solubilization process of which is intended to be facilitated. In fact, as the studies by Waithelm U et al, (“Effects of Saponins on the Water Solubility of Different Model Compounds”, Planta Med 2001, 67) show, the solubility of a given molecule can be increased in the presence of a given saponin but at the same time reduced if a different saponin is used.

[0063] As will be explained in more detail in the following experimental section, the present inventors have surprisingly found that a saponin belonging to the triterpenoid class of saponins, more specifically the asiaticoside saponin, is capable of exerting a marked solubilizing activity on the galunisertib molecule, allowing stable suspensions of this molecule in water to be obtained without the need to add additional excipients.

[0064] Within the scope of the present description, the term “stable suspension” refers in particular to an aqueous formulation in which phase separation and / or precipitation of at least one of the two components (e.g., asiaticoside or galunisertib) does not occur at the end of the process of preparation of said aqueous formulation in the absence of additional solubilizing excipients.

[0065] As will be apparent from the examples below, the use of the asiaticoside saponin as an excipient in the pharmaceutical composition of the invention advantageously allows the known low aqueous solubility of the active ingredient galunisertib (0.0112 mg / ml, SOURCE: DrugBank) to be significantly increased, even up to 80 times, thereby allowing for a concentration of galunisertib in solution of approximately 0.9 mg / ml. The inventors surprisingly found that a considerable increase in solubility of the above-mentioned active ingredient required the use of a small quantity of the saponin asiaticoside, in a galunisertib / asiaticoside weight ratio of only 1:1.8.

[0066] Further studies carried out by the present inventors have also shown that the critical micelle concentration (CMC) of the asiaticoside molecules in the pharmaceutical composition for use according to the invention has a value of approximately 30.7 ± 6.5 pg / ml, thus indicating a clear tendency of said molecules to aggregate in water into three-dimensional micellar structures (see, for example, Example 4).

[0067] The term “critical micelle concentration”, as used herein, refers to the concentration value of a solution of molecules at which a number of said molecules cluster into micelles.

[0068] Therefore, in one embodiment of the pharmaceutical composition for use according to the invention, the TGF-beta receptor inhibitor is enclosed within nanoparticles consisting of triterpenoid saponin molecules arranged in the form of spherical micelles.

[0069] The term “in the form of a spherical micelle”, as used herein, refers to a spherical structure resulting from the clustering of the triterpenoid saponin molecules as defined above in such a way that the aglycone is located inside the spherical structure and the sugar portion of the saponin is oriented on the outside of said structure.

[0070] Preferably, the triterpenoid saponin molecules according to the invention are selected from the group consisting of asiaticoside, astragaloside IV, ginsenoside Rgl, glycyrrhizin, and any combination thereof.

[0071] In one embodiment according to the invention, the nanoparticles consist of asiaticoside molecules arranged in the form of spherical micelles.

[0072] In another embodiment, the nanoparticles consist of glycyrrhizin molecules arranged in the form of spherical micelles. In a further embodiment, the pharmaceutical composition for use according to the invention comprises nanoparticles consisting of asiaticoside and glycyrrhizin molecules arranged in the form of spherical micelles.

[0073] According to one embodiment of the invention, the nanoparticles as defined above have a size distribution ranging from 10 to 1,000 nm, preferably from 50 to 800 nm, even more preferably from 150 to 650 nm.

[0074] According to another embodiment, the zeta potential of the nanoparticles in the pharmaceutical composition for use according to the invention ranges from -5 mV to -40 mV.

[0075] The term “zeta potential”, as used herein, refers to the charge that develops at the interface between a solid surface and the liquid in which it is immersed.

[0076] Particularly preferred is a pharmaceutical composition for use according to the invention comprising the TGF-beta inhibitor galunisertib, wherein said inhibitor is enclosed within nanoparticles consisting of asiaticoside saponin molecules arranged in the form of spherical micelles.

[0077] Another particularly preferred pharmaceutical composition for use according to the invention comprises the TGF-beta inhibitor vactosertib, wherein said inhibitor is enclosed within nanoparticles consisting of glycyrrhizin molecules arranged in the form of spherical micelles.

[0078] The selection of the most suitable encapsulation method for use within the scope of the present invention falls well within the skills of those of ordinary skill in the art.

[0079] As explained above, the requirement of a small amount of triterpenoid saponin, e.g., asiaticoside and / or glycyrrhizin, as the solubilizing excipient, advantageously allows for the formation of nanoparticles having a high content of the TGF-beta inhibitory agent, equal to at least 55% by weight of the total weight of the nanoparticle. In one embodiment of the composition according to the invention, said TGF-beta receptor inhibitor, e.g., galunisertib and / or vactosertib, and said triterpenoid saponin molecules, e.g., asiaticoside and / or glycyrrhizin, are present in the nanoparticles in a molar ratio ranging from 8: 1 to 0.1: 1, for example, in a molar ratio ranging from 7: 1 to 0.5:1, or from 6: 1 to 1: 1, or from 5:1 to 0.5:1. Preferably, said molar ratio is 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, or 1:1.

[0080] This property of the nanoparticles in the pharmaceutical composition for use according to the invention is completely unexpected in the light of the teachings of the prior art. By way of example, the paper by Tramontane C et al. (Tramontane C. et al, “Micro fluidic- Assisted Production of Gastro-Resistant Active-Targeted Diatomite Nanoparticles for the Local Release of Galunisertib in Metastatic Colorectal Cancer Cells”, Adv Healthc Mater. 2022, 12 (6)) describes the encapsulation of galunisertib in nanoparticle systems that require the use of an excess of excipient, equal to 40 times the mass of galunisertib, resulting in an extremely low loading of the active ingredient, between 4.0 and 4.5% by weight of the total weight of the nanoparticle. Similarly, the paper by Sun X et al. (Sun X et al. “Acid-switchable nanoparticles induce self-adaptive aggregation for enhancing antitumor immunity of natural killer cells”, Acta Pharmaceutica Sinica B, 2023, 13(7) 3093-3105) teaches a high excipient / galunisertib ratio for the formation of certain nanoparticles, equal to 16:1, and a galunisertib content in the nanoparticles thus obtained that does not exceed 4.5% of the total weight of the nanoparticle.

[0081] The pharmaceutical composition for use according to the invention may also comprise additional pharmaceutically acceptable ingredients, such as, for example, stabilizing agents, such as mannitol, lubricating agents, such as magnesium stearate, buffer agents, such as citrate buffer, antioxidant agents, such as EDTA, and / or preservative agents, such as methyl hydroxybenzoate.

[0082] Of course, the selection of suitable carriers, excipients and / or diluents falls within the skills of those of ordinary skill in the art. The selection of the dose of the active ingredient and the dosage regimen also fall within the skills of those of ordinary skill in the art, and the selection thereof depends on several factors, such as for example the age and weight of the patient, as well as the degree of progression of the disease.

[0083] The present invention is also based on the surprising finding by the inventors that the use of a composition according to the invention, in particular a composition comprising the active ingredient galunisertib in combination with the excipient asiaticoside, in experimental models of pulmonary fibrosis results in a significant reduction in the expression levels of numerous primary biomarkers of said disease. In addition, as shown in Figures 5, 6 and 7, the extent of the reduction of said expression levels as measured by the inventors after treatment of the cells with the composition according to the invention is significantly higher than that observed after the use of galunisertib or asiaticoside alone, which represents a clear indication of the existence of a marked synergistic effect of the aforementioned two molecules. In accordance with the results of the in vitro experiments, the marked antifibrotic and anti-inflammatory activity exhibited by the composition of the invention was also subsequently validated in vivo by using a murine model of idiopathic pulmonary fibrosis (IPF) induced by bleomycin (Figures 8 - 10).

[0084] In light of the results illustrated above, the pharmaceutical composition according to the invention is therefore particularly suitable for therapeutic use for the treatment of fibrotic lung diseases, in particular a fibrotic lung disease selected from the group consisting of idiopathic pulmonary fibrosis (IPF), non-specific interstitial pneumonia (NSIP), chronic hypersensitivity pneumonitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), pulmonary sarcoidosis, pneumoconiosis, cryptogenic organizing pneumonia, interstitial lung disease associated with autoimmune diseases, and any combination thereof. Idiopathic pulmonary fibrosis (IPF) is particularly preferred.

[0085] One parameter used for the diagnosis of idiopathic pulmonary fibrosis is the forced vital capacity (FVC). In healthy subjects, the mean values are 4,800 ml for men and 3,200 ml for women. Patients with idiopathic pulmonary fibrosis experience a reduction in the FVC parameter.

[0086] Another useful parameter to assess IPF disease is the forced expiratory volume (FEV 1 or VEMS), which indicates the volume of air exhaled during the first second of a maximum forced expiration, and is expressed as a percentage of the expected (100%). In healthy subjects, FEV1 is between 80% and 120%. In patients with IPF, the decrease in FEV1 is proportional to the decrease in the FVC value.

[0087] In patients with pulmonary fibrosis undergoing therapeutic treatment, an increase in FVC values, FEV1 values, or both, indicates the effectiveness of the treatment. Usually, in the clinical practice, the ratio between FEV1 and FVC measurements (expressed as a percentage) is assessed.

[0088] The preferred embodiments described above can be combined with each other as required, and the implementation of these combinations falls within the skills of the person skilled in the art.

[0089] The examples that follow are provided for illustration purposes and do not limit the scope of the invention as defined in the appended claims.

[0090] EXAMPLES

[0091] Example 1: Preparation of compositions according to the invention comprising the active ingredient galunisertib and the excipient asiaticoside

[0092] For the preparation of a composition according to the invention, the TGF-beta inhibitor galunisertib and the saponin asiaticoside were dissolved in an ethanol / water mixture and subsequently rapidly injected into a predetermined volume of aqueous solvent. Once the injection phase was completed, the organic solvent was removed by vacuum evaporation, and the final volume of water was adjusted based on the desired concentration of the above molecules. In a particular embodiment, 2.25 mg galunisertib and 4.05 mg asiaticoside (weight ratio 1:1.8) were dissolved in 500 pL of an ethanol / water mixture (ratio 90: 10). The resulting solution was then injected into 2.5 ml of milliQ water (4°C) under stirring. The resulting suspension was then subjected to vacuum evaporation to remove the ethanol and part of the water up to a final volume of 2.5 ml in order to obtain a final concentration of 0.9 mg / ml galunisertib and 1.62 mg / ml asiaticoside. The sample was then centrifuged for 10 minutes at 4000 RPM to remove any aggregates, and the supernatant consisting of a suspension of galunisertib and asiaticoside, more particularly a suspension of nanoparticles consisting of asiaticoside molecules and containing on the inside the active ingredient galunisertib, was collected and stored at 4°C.

[0093] Following the above method, a second composition according to the invention was prepared by varying the weight ratio between galunisertib and asiaticoside. Specifically, a composition with a galunisertib-asiaticoside weight ratio of 1:0.5 was prepared.

[0094] Table 1 below gives the values of the chemical-physical parameters of the nanoparticles of the compositions produced, measured immediately after their preparation.

[0095] Table 1

[0096] Zeta

[0097] Galunisertib / asiaticoside pg / ml Size (nm+SD) potential PDI

[0098] weight ratio Galunisertib (mV)

[0099] 1:1.8 253.7 + 83.0 -19.8 + 6.0 0.568 900 + 50

[0100] 1:0.5 428.5 + 198.0 -16.7 + 8.1 0.573 640 + 80

[0101]

[0102] Example 2: Preparation of compositions according to the invention comprising vactosertib and the excipient glycyrrhizin

[0103] The present inventors prepared an additional composition according to the invention using the TGF-beta inhibitor vactosertib and the saponin glycyrrhizin. For this purpose, the above compounds were dissolved in ethanol and subsequently rapidly injected into a predetermined volume of aqueous solvent. Once the injection phase was completed, the organic solvent was removed by vacuum evaporation, and the final volume of water was adjusted based on the desired concentration of the above molecules. In a particular embodiment, 0.20 mg vactosertib and 4.05 mg glycyrrhizin (weight ratio 1:20.25) were dissolved in 500 pL of an ethanol / water mixture (ratio 80:20). The resulting solution was then injected into 2.5 ml of milliQ water under stirring. The resulting suspension was then subjected to vacuum evaporation to remove the ethanol and part of the water up to a final volume of 2.5 ml in order to obtain a final concentration of 0.08 mg / ml vactosertib and 1.62 mg / ml glycyrrhizin.

[0104] The sample was then centrifuged for 10 minutes at 4000 RPM to remove any aggregates, and the supernatant consisting of a suspension of vactosertib and glycyrrhizin, more particularly a suspension of nanoparticles consisting of glycyrrhizin molecules and containing on the inside the active ingredient vactosertib, was collected and stored at 4°C.

[0105] Table 2 below gives the values of the chemical-physical parameters of the nanoparticles of the composition produced, measured immediately after its preparation.

[0106] Table 2

[0107] Vactosertib / Zeta

[0108] Size pg / ml glycyrrhizin potential PDI

[0109] (nm+SD) vactosertib weight ratio (mV)

[0110] 1:20.25 388.9 + 17.7 -10.9 + 5.0 0.411 146.5 + 12.0

[0111]

[0112] Example 3: Chemical-physical characterization of the composition for use according to the invention

[0113] The present inventors carried out dedicated tests with the aim of characterizing the composition object of the invention according to various physical-chemical properties as described below. The tests carried out by the inventors also concerned compositions comprising nanoparticles consisting of asiaticoside molecules and containing on the inside the active ingredient galunisertib.

[0114] 1. Apparent solubility Apparent solubility refers to the solubility of a solute in a solvent, as measured under certain experimental conditions. In this specific case, the apparent solubility represents the range of concentrations at which stable aqueous suspensions comprising galunisertib and asiaticoside can be obtained. Stable suspensions refer to aqueous suspensions comprising galunisertib and asiaticoside and having a concentration higher than the aqueous solubility limit of galunisertib, where no phase separation and / or precipitate is observed at the end of the method of preparation of the suspension, and where stability, as described, is achieved without the aid of any additional stabilizing and / or solubilizing agent.

[0115] 2. Size of the nanoparticles consisting of asiaticoside molecules and containing galunisertib on the inside

[0116] The analysis was performed using the Zeta-sizer Nano Z instrument (Malvern Instruments, Malvern, UK). Measurements were taken at 25°C in milliQ water. The technique used to measure the diameter of the nanoparticles is called Dynamic Light Scattering (DLS). This technique allows the hydrodynamic diameter of nanoparticles dispersed in a liquid to be measured. In short, the sample is illuminated by a laser beam, and changes in intensity of the light scattered by the sample are measured as a function of time. The intensity changes measured by the detector are generated by the Brownian motion of the particles. At the same temperature and viscosity, small particles move rapidly creating rapid changes in the scattering intensity, whereas larger particles move more slowly creating slow changes in the intensity.

[0117] 3. Zeta potential of the nanoparticles consisting of asiaticoside molecules and containing galunisertib on the inside

[0118] The analysis was performed using the Zeta-sizer Nano Z instrument (Malvern Instruments, Malvern, UK). All samples were analysed at 25°C in milliQ water. The zeta potential of the nanoparticles of the invention was determined by measuring the electrophoretic mobility of the particles suspended in water. To measure the electrophoretic mobility of the particles, an electric field was applied between the electrodes of the measuring cell which holds the sample and is illuminated by a laser beam. The charged particles move towards the opposite- sign electrode, creating a change in frequency of the light scattered by the sample, which is directly proportional to the electrophoretic mobility

[0119] Example 4: Assessment of the critical micelle concentration (CMC)

[0120] The critical micelle concentration (CMC) is the concentration at which a number of monomers cluster, leading to the formation of micelles.

[0121] The multi-angle dynamic light scattering technology based on the measurement of the scattering intensity was used to determine the CMC of the composition of the invention relating to the asiaticoside molecules.

[0122] The scattering intensity is the intensity of scattered light as a result of the interaction of the light beam with the nanoparticles, which is a function of the concentration of monomers in solution. As the monomers cluster to form micelles, the scattering intensity increases significantly.

[0123] The diagram in Figure 1, showing the correlograms obtained from the analysis of the asiaticoside sample in milliQ water at 25°C at each tested concentration, shows the trend of the intercept of the correlation function, which increases as the tested concentration increases. The horizontal line represents the CMC limit of 0.8 g2-l for the formation of colloidal structures / CMC.

[0124] The diagram in Figure 2 shows the scattering intensity (“derived count rate”) as a function of the concentration of asiaticoside nanoparticles, with its trend line.

[0125] The CMC value appears to be 30.7 ± 6.5 pg / ml from the combination of the test results of the correlograms and scattering intensities, as a function of the asiaticoside concentrations tested.

[0126] Example 5: In vitro assessment of the effects of galunisertib on transforming growth factor beta (TGF-beta) inhibition As is known, galunisertib is a potent inhibitor of the transforming growth factor beta (TGF-beta) receptor. In order to investigate possible activity of this active ingredient on the fibrotic process, the inventors carried out in vitro studies on primary human pulmonary fibroblast (HPF) cells.

[0127] Two separate experiments were performed, treating HPF cells with two different concentrations of TGF-beta (1 and 5 nanograms / ml, respectively) in order to induce differentiation of fibroblasts into myofibroblasts. For each experiment, cells were treated with three increasing concentrations of galunisertib (0.5 - 2.5 - 10 pM) over a 72-hour period. At the end of the treatment, the cell proliferation index was assessed by a method that determines the number of viable cells by quantifying the levels of Adenosine Triphosphate (ATP). The kit used (CellTiter-Glo®, Promega) generates a luminescent signal proportional to the number of viable cells. The viability of HPF cells treated with TGF-beta and galunisertib was compared with HPF cell samples not stimulated with TGF-beta and with cell samples that, after TGF-beta stimulation, did not undergo treatment with galunisertib.

[0128] As shown in the diagrams in Figure 3A (cells treated with 1 ng / ml TGF-beta) and 3B (cells treated with 5 ng / ml TGF-beta), showing the measurement of cell viability as a function of treatment, HPF cells treated with TGF-beta alone at both concentrations exhibit a reduction in cell proliferation ascribable to differentiation of fibroblasts into myofibroblasts. In contrast, HPF cells stimulated with TGF-beta and subsequently treated with galunisertib exhibit a cell proliferation index comparable to that of cells not treated with TGF-beta. This result indicates the ability of galunisertib to block TGF-beta-induced differentiation of fibroblasts into myofibroblasts. In the diagrams in Figures 3 A and 3B, statistical significance between the different conditions was determined by analysis of variance (ordinary one-way ANOVA or Krustal Wallis test according to the data distribution) followed by Dunnett’s or Dunn’s multi comparison post-hoc test, respectively (*:p < 0.05; **:p < 0.01; ***:p < 0.001). Data are expressed as mean ± SEM.

[0129] Example 6: Assessment of in vitro toxicity of asiaticoside on primary human fibroblast cells (HPF) The present inventors conducted an in vitro experiment to assess the toxicity of the asiaticoside molecule on primary human pulmonary fibroblast (HPF) cells. Specifically, HPF cells were treated with increasing concentrations of asiaticoside (1, 10, 25, 50, 100 pg / ml) for 24 hours. Cell viability was measured according to the method given in Example 4. As shown in the diagrams in Figure 4, the asiaticoside concentrations tested did not result in a significant reduction in cell viability compared to the control (cells not treated with asiaticoside), except for the concentration of 100 pg / ml.

[0130] Example 7: In vitro inhibition of a-SMA ("alpha smooth muscle actin”) expression by asiaticoside, galunisertib and a composition according to the invention

[0131] One of the main markers of differentiation of fibroblasts into myofibroblasts is the actin a-SMA. This is a contractile protein that promotes migration and contraction of myofibroblasts, resulting in tissue stiffness and consequent lung dysfunction (Kolanko E et al., “The evolution of in vitro models of lung fibrosis: promising prospects for drug discovery”, European Respiratory Review, 2024, 33).

[0132] The present inventors carried out dedicated studies to assess the ability of galunisertib, asiaticoside, and / or a composition comprising a combination of these compounds, in particular a composition comprising nanoparticles consisting of asiaticoside molecules and containing galunisertib, to reduce the expression of the a-SMA protein. For these studies, the inventors employed an in vitro model of primary human pulmonary fibroblast (HPF) cells. In short, HPF cells were treated for 3 days with TGF-beta at a concentration of 10 ng / ml, to induce a-SMA expression, and then with the test molecules (asiaticoside, galunisertib, and a composition comprising galunisertib and asiaticoside). Asiaticoside was tested at concentrations of 7 and 50 pg / ml, galunisertib at concentrations of 0.5 and 10 pM, and the combination of galunisertib and asiaticoside at the above concentrations, both molecules being associated in increasing manner. Quantification of a-SMA protein expression levels was performed by quantitative PCR. More specifically, total RNA was isolated from the cells subjected to the above treatments and from the untreated control sample using a commercial kit, following the manufacturer’s instructions. The quality of the purified RNA was verified to ensure its suitability for subsequent experimental applications. The resulting RNA samples were subjected to a reverse transcription reaction for the synthesis of the complementary DNA (cDNA) using a commercial kit. Up to 2 pg of total RNA were converted to single- stranded cDNA in a final reaction volume of 20 pL, using random primers. A quantitative PCR (qPCR) reaction was then performed using the cDNA samples as the template and SYBR™ Green fluorescent dye to detect the double- stranded DNA amplification products. Cycle threshold (Ct) values were normalized to a reference gene (housekeeping gene), and relative expression levels were calculated using the AACt method. The results are expressed as relative change in expression (RQ, relative quantification) compared to a reference sample (calibrator).

[0133] As can be seen from Figures 5 A and 5B, treatment with asiaticoside alone or galunisertib alone at both concentrations results in a reduction in a-SMA protein expression levels. This effect is also observed after treatment of the cells with the composition comprising galunisertib and asiaticoside (nanoparticles consisting of asiaticoside molecules and containing galunisertib). Importantly, the above-mentioned composition, already at the lowest concentrations of asiaticoside and galunisertib, results in a greater reduction in a-SMA levels than that obtained by treating the cells with galunisertib alone.

[0134] Example 8: In vitro inhibition of fibronectin expression by asiaticoside, galunisertib and a composition according to the invention

[0135] Fibronectin is a glycoprotein involved in the pathogenesis of pulmonary fibrosis. It contributes to collagen deposition, tissue remodelling, and is one of the markers of the epithelial-mesenchymal transition (EMT). EMT is a phenomenon in which epithelial cells lose their apical-basolateral characteristics and acquire a mesenchymal phenotype resulting in increased migratory capacity and extracellular matrix production.

[0136] For the purpose of assessing the ability of asiaticoside, galunisertib, and a composition comprising a combination of these compounds, more specifically a composition comprising nanoparticles consisting of asiaticoside molecules and containing galunisertib, to reduce fibronectin expression, the inventors carried out in vitro studies on human bronchial epithelial cells (HBEC). Specifically, HBEC cells were stimulated with TGF-beta at a concentration of 10 ng / ml to induce fibronectin expression and subsequently treated with asiaticoside (50 pg / ml), galunisertib (10 pM), and the combination of galunisertib and asiaticoside (10 pM and 50 pg / ml) for 72 hours, respectively. The quantification of fibronectin expression in the cells subjected to the different treatments and in the untreated sample was carried out by quantitative PCR following the experimental protocol described in Example 7 with reference to the analysis of a-SMA actin expression.

[0137] As can be seen from Figure 6, treatment with asiaticoside alone results in increased fibronectin expression. In contrast, treatment with galunisertib alone results in reduced fibronectin expression. Surprisingly, when using galunisertib in combination with asiaticoside, especially nanoparticles consisting of asiaticoside molecules and containing galunisertib, there is a greater reduction in fibronectin levels than with galunisertib alone, indicating a synergistic effect. This synergistic effect is completely unexpected considering that asiaticoside alone shows a detrimental effect on fibronectin levels.

[0138] Example 9: In vitro inhibition of vimentin expression by asiaticoside, galunisertib and a composition according to the invention

[0139] Vimentin is a protein found in mesenchymal cells and plays an important role in the fibrotic process. Its expression results in increased cellular invasiveness and increased scar tissue (Surolia R et al. “Vimentin intermediate filament assembly regulates fibroblast invasion in fibrogenic lung injury”, JCI Insight, 20194;4(7)). The vimentin protein is also involved in the epithelial-mesenchymal transition (EMT) process.

[0140] Dedicated experiments were carried out in order to assess the ability of asiaticoside, galunisertib, and a composition comprising a combination of these compounds, more specifically a composition comprising nanoparticles consisting of asiaticoside molecules and containing galunisertib, to reduce vimentin expression. For these experiments, the inventors employed an in vitro model of human bronchial epithelial cells (HBEC). More specifically, HBEC cells were treated for 72 hours with TGF-beta at a concentration of 10 ng / ml to induce vimentin expression. Subsequently, the cells were placed in contact with asiaticoside, galunisertib, and the composition as defined above. Asiaticoside was tested at a concentration of 50 pg / ml, galunisertib at a concentration of 10 pM, and galunisertib in combination with asiaticoside at the concentrations of 10 pM and 50 pg / ml, respectively. The quantification of vimentin expression in the cells subjected to the different treatments and in the untreated sample was carried out by quantitative PCR following the experimental protocol described in Example 7 with reference to the analysis of a-SMA actin expression.

[0141] As can be seen from Figure 7, treatment with asiaticoside alone results in increased vimentin expression, while the expression levels of this protein do not change significantly after treatment with galunisertib alone, remaining comparable to those observed after induction with TGF-beta. In this case, too, as already surprisingly observed for the marker fibronectin, combining the use of galunisertib with the excipient asiaticoside results in a significant reduction in vimentin levels. This undoubtedly indicates the existence of a synergistic effect between the aforementioned active ingredient and the aforementioned excipient, which is completely unexpected in light of the results described above obtained with galunisertib alone or with asiaticoside alone.

[0142] Example 10: In vivo analysis of the therapeutic efficacy of a composition according to the invention

[0143] An in vivo test was performed on a mouse model of bleomycin-induced idiopathic pulmonary fibrosis (IPF) to assess the therapeutic efficacy of a pharmaceutical composition according to the invention. The aim of the study was to analyse the effect of the administration of the pharmaceutical composition of the invention on the cytokine expression profile, inflammation in the bronchoalveolar lavage fluid (BAEF), and collagen deposition in the lung.

[0144] Pulmonary fibrosis was induced in the animals by intratracheal administration of bleomycin (2.5 mg / kg body weight) on Day 0. From Day 7 to Day 21, the animals were treated by administration of a composition according to the invention comprising nanoparticles consisting of asiaticoside molecules and containing galunisertib. For each animal, the total daily dosage of galunisertib was 2.1 mg per kg of animal body weight, while the daily dosage of asiaticoside was 3.8 mg per kg of animal body weight.

[0145] In order to determine the effectiveness of the treatment, the study was conducted on three different experimental groups:

[0146] - Untreated healthy animals (“NAIVE group”);

[0147] - Untreated diseased animals (“BLM group”);

[0148] - Diseased animals treated with a composition according to the invention comprising nanoparticles consisting of asiaticoside molecules and containing galunisertib as described above (“BLM group + nanoparticles”).

[0149] On Day 22, the animals were sacrificed and the bronchoalveolar lavage fluid (BALF) was collected by performing three successive lung lavages with ice-cold sterile phosphate buffered saline (PBS) using an endotracheal tube. For each animal tested, the supernatants of the three successive lavages were combined and used for the analysis of the cytokines interleukin 6 (IL-6), interleukin 11 (IL-11), interferon gamma (IFN-y) and transforming growth factor-P (TGF-P), using mouse-specific ELISA kits (Invitrogen™). The cell pellets were resuspended in PBS and subjected to total and differential cell counts (macrophages, lymphocytes, neutrophils). Hydroxyproline content analysis was performed on lung samples using the “Hydroxyproline assay” kit (Sigma- Aldrich).

[0150] As shown by the diagrams in Figure 8, the analysis of the levels of inflammatory cytokines determined in the animals following the treatment described above gave the following results:

[0151] IL-6 and IL-11: Treatment with the composition of the invention resulted in a significant reduction in the mean concentration levels of these cytokines compared to the diseased group, equal to 38% (IL-6, Figure 8A) and 57% (IL-11, Figure 8B), respectively;

[0152] IFN-y: A marked decrease in the levels of this cytokine was observed in the treated group, equal to 29%, compared to the diseased group (Figure 8C);

[0153] TGF-P: A highly significant mean reduction in concentration, equal to 37%, was also found for the levels of this fibrogenic cytokine after treatment with the composition of the invention (Figure 8D). The results of the cytological analysis of BALF, shown in Figure 9, demonstrated that treatment with the composition of the invention resulted in a significant reduction in the following parameters, compared to the untreated diseased group: 36% reduction in total cell count (Figure 9A), 28% reduction in the number of macrophages (Figure 9B), 36% reduction in the number of lymphocytes (Figure 9C), and 58% reduction in the number of neutrophils (Figure 9D). These data clearly indicate a marked anti-inflammatory effect of the composition of the invention at the pulmonary level.

[0154] In addition, the results of the analysis for hydroxyproline content, carried out to assess collagen deposition in the lung, shown in the diagram in Figure 10, indicate a 26% reduction in the concentration of this protein in the treated group of animals compared to the diseased group.

[0155] The study described above shows that, in a murine model of idiopathic pulmonary fibrosis, treatment with a composition of the invention comprising nanoparticles consisting of asiaticoside molecules and containing galunisertib achieves significant therapeutic effects. In particular, this treatment resulted in a modulation, i.e., reduction, of the expression of pro-inflammatory and pro-fibrotic cytokines, as well as a decrease in inflammatory cell infiltrate in the bronchoalveolar lavage fluid (BALF) and a reduction in hydroxyproline content in the lungs.

Claims

CLAIMS1. A pharmaceutical composition comprising a TGF-beta receptor inhibitor selected from the group consisting of Galunisertib (LY2157299), Vactosertib (EW-7197), Itacnosertib (TP-0184), and Repsox (E-616452), and any combination thereof, and at least one pharmaceutically acceptable carrier, excipient and / or diluent, wherein the at least one pharmaceutically acceptable excipient comprises triterpenoid saponin molecules, for use in the therapeutic treatment of a fibrotic lung disease.

2. The pharmaceutical composition for use according to claim 1, which is in a pharmaceutical form for administration via inhalation.

3. The pharmaceutical composition for use according to claim 2, wherein said pharmaceutical form is an inhalable dry powder.

4. The pharmaceutical composition for use according to any one of claims 1 to 3, wherein the TGF-beta receptor inhibitor is enclosed within nanoparticles consisting of triterpenoid saponin molecules arranged in the form of spherical micelles.

5. The pharmaceutical composition for use according to any one of claims 1 to 4, wherein said triterpenoid saponin molecules are selected from the group consisting of asiaticoside, astragaloside IV, ginsenoside Rgl, glycyrrhizin, and any combination thereof.

6. The pharmaceutical composition for use according to claim 4 or 5, wherein said TGF-beta receptor inhibitor and said triterpenoid saponin molecules are present in the nanoparticles in a molar ratio ranging from 8: 1 to 0.1: 1.

7. The pharmaceutical composition for use according to any one of claims 1 to 6, further comprising a therapeutic agent selected from the group consisting of corticosteroids, antifibrotic agents, phosphodiesterase inhibitors, immunosuppressive agents, and any combination thereof.

8. The pharmaceutical composition for use according to any one of claims 1 to 7, wherein the fibrotic lung disease is selected from the group consisting of idiopathic pulmonary fibrosis (IPF), non-specific interstitial pneumonia (NSIP), chronic hypersensitivity pneumonitis, systemic sclerosis-associated interstitial lung disease (SSc-ILD), pulmonary sarcoidosis, pneumoconiosis, cryptogenic organizing pneumonia, interstitial lung disease associated with autoimmune diseases, and any combination thereof.

9. The pharmaceutical composition for use according to any one of claims 1 to 8, wherein the treatment is a therapy that inhibits the differentiation of lung fibroblasts into myofibroblasts in a subject affected by a fibrotic lung disease.

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