Dry powder compositions of treprost prodrugs and methods of use thereof

Abstract

The present disclosure provides a dry powder composition of a treprost prodrug and a method of treating pulmonary hypertension (e.g., pulmonary arterial hypertension), portal pulmonary hypertension, or pulmonary fibrosis in a patient in need thereof. The dry powder composition comprises (a) from about 0.1 wt% to about 3 wt% of a compound of formula (I): or an enantiomer, diastereoisomer or pharmaceutically acceptable salt thereof, (b) from about 0.01 wt% to about 3 wt% of DSPE-PEG2000, (c) from about 10 wt% to about 50 wt% of leucine, and the balance (d) a sugar selected from the group consisting of trehalose and mannitol. The total of (a), (b), (c) and (d) is 100 wt%, and R1 is tetradecyl, pentadecyl, hexadecyl, heptadecyl or octadecyl. The method comprises administering a therapeutically effective amount of the dry powder composition to the lung of the patient by inhalation via a dry powder inhaler. In certain compositions and methods provided herein, R1 is a hexadecyl group, such as a linear hexadecyl group. # imgabs0 #

Description

This application is a divisional application of an invention patent application with an application date of April 28, 2020, application number 202080047360.0 (international application number PCT / US2020 / 030282), and name “Dry powder composition of treprostinil prodrug and method of use thereof”. CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application Serial No. 62 / 840,186, filed on April 29, 2019, the disclosure of which is incorporated herein by reference in its entirety. Background Art

[0002] Pulmonary hypertension (PH) is characterized by abnormally high blood pressure in the pulmonary vessels. It is a progressive, fatal disease that leads to heart failure and can occur in the pulmonary arteries, pulmonary veins, or pulmonary capillaries. Symptomatic patients experience shortness of breath, dizziness, fainting, and other symptoms, all of which are aggravated by exertion. There are multiple causes and may be of unknown origin, idiopathic, and may lead to high pressure in other systems, for example, portopulmonary hypertension in which patients suffer from both portal hypertension and pulmonary hypertension.

[0003] The World Health Organization (WHO) has divided pulmonary hypertension into five groups. The first group is called pulmonary arterial hypertension (PAH), and includes PAH of unknown cause (idiopathic), hereditary PAH (i.e., familial PAH or FPAH), PAH caused by drugs or toxins, and PAH caused by conditions such as connective tissue disease, HIV infection, liver disease, and congenital heart disease. The second group of pulmonary hypertension is characterized as pulmonary hypertension associated with left heart disease. The third group of pulmonary hypertension is characterized as PH associated with lung disease (such as chronic obstructive pulmonary disease and interstitial lung disease) and PH associated with sleep-related breathing disorders (e.g., sleep apnea). The fourth group of PH is PH caused by chronic thrombosis and / or embolic disease, such as PH caused by blood clots or coagulation disorders in the lungs. The fifth group includes PH caused by other disorders or conditions such as blood disorders (e.g., polycythemia vera, essential thrombocythemia), systemic disorders (e.g., sarcoidosis, vasculitis), and metabolic disorders (e.g., thyroid disease, glycogen storage disease).

[0004] Pulmonary arterial hypertension (PAH) affects approximately 200,000 people worldwide, with approximately 30,000-40,000 of those patients in the United States. PAH patients experience constriction of the pulmonary arteries, which causes increased pressure in the pulmonary arteries, making it difficult for the heart to pump blood to the lungs. Patients suffer from shortness of breath and fatigue, which often severely limits the ability to perform physical activities.

[0005] The New York Heart Association (NYHA) classifies PAH patients into four functional categories to assess the severity of the disease. PAH patients in Class I, as classified by the NYHA, have no limitations on physical activity because ordinary physical activity does not cause excessive dyspnea or fatigue, chest pain, or near syncope. PAH patients in Class II, as classified by the NYHA, have slight limitations on physical activity. These patients are comfortable at rest, but ordinary physical activity causes excessive dyspnea or fatigue, chest pain, or near syncope. PAH patients in Class III, as classified by the NYHA, have significant limitations on physical activity. Although comfortable at rest, PAH patients in Class III experience excessive dyspnea or fatigue, chest pain, or near syncope due to activities less than ordinary physical activity. PAH patients in Class IV, as classified by the NYHA, cannot perform any physical activity without symptoms. PAH patients in Class IV may experience dyspnea and / or fatigue at rest, and any physical activity increases discomfort. PAH patients in Class IV often show signs of right heart failure.

[0006] Patients with PAH are treated with endothelin receptor antagonists (ERAs), phosphodiesterase type 5 (PDE-5) inhibitors, guanylate cyclase stimulators, prostaglandins (e.g., prostacyclins), or a combination thereof. ERAs include ambrisentan Sitaxsentan, Bosentan and macitentan PDE-5 inhibitors used to treat PAH include sildenafil and tadalafil Prostaglandins used to treat PAH include iloprost, epoprostenol, and treprostinil An approved guanylate cyclase stimulator is riociguat Additionally, patients are often treated with a combination of the above-mentioned compounds.

[0007] Portopulmonary hypertension (PPH) is defined as the coexistence of portal hypertension and pulmonary hypertension and is a serious complication of liver disease. The diagnosis of PPH is based on hemodynamic criteria: (1) portal hypertension and / or liver disease (clinical diagnosis - ascites / varicose veins / splenomegaly), (2) resting mean pulmonary artery pressure >25 mmHg, and (3) pulmonary vascular resistance >240 dynes / cm 5 , (4) Pulmonary artery occlusive pressure <15 mmHg or transpulmonary gradient >12 mmHg. PPH is a serious complication of liver disease and is present in 0.25% to 4% of patients with cirrhosis. Today, PPH is a comorbidity in 4%-6% of patients referred for liver transplantation.

[0008] Pulmonary fibrosis is a respiratory disease in which scarring forms in the lung tissue, leading to severe breathing problems. Scarring, the accumulation of excess fibrous connective tissue, causes the walls to thicken and leads to a reduction in the oxygen supply to the blood. As a result, patients with pulmonary fibrosis suffer from permanent shortness of breath. In some patients, a specific cause of the disease can be diagnosed, while in others no possible cause can be identified, a condition known as idiopathic pulmonary fibrosis.

[0009] The present invention addresses the need for new treatment options for pulmonary hypertension (PH), including pulmonary arterial hypertension (PAH), portopulmonary hypertension (PPH), and pulmonary fibrosis by providing dry powder compositions of Treprostinil prodrugs useful for pulmonary administration and methods of administering the same to patients in need thereof. Summary of the invention

[0010] In one aspect, the present disclosure relates to a dry powder composition. The dry powder composition comprises (a) about 0.1 wt % to about 3 wt % of a compound of formula (I): or an enantiomer, a diastereomer or a pharmaceutically acceptable salt thereof, wherein R 1 is tetradecyl, pentadecyl, hexadecyl, heptadecyl or octadecyl; (b) about 0.01 wt% to about 3 wt% of distearoylphosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000); (c) about 10 wt% to about 50 wt% of leucine; and the rest is (d) a sugar selected from trehalose and mannitol. The total of (a), (b), (c) and (d) is 100 wt%.

[0011] In one embodiment, R 1 In other embodiments, R 1 It is a linear tetradecyl group.

[0012] In one embodiment, R 1 In other embodiments, R 1 It is a linear pentadecyl group.

[0013] In one embodiment, R 1 In other embodiments, R 1 It is a linear heptadecanyl group.

[0014] In one embodiment, R 1 In other embodiments, R 1 It is a linear octadecyl group.

[0015] In one embodiment, R 1 In other embodiments, R1 It is a linear hexadecyl group.

[0016] In one embodiment, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 0.5 wt% to about 2 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.05 wt% to about 2 wt% of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In other embodiments, the DSPE-PEG2000 is present at about 0.15 wt% to about 1.4 wt% of the total weight of the dry powder composition. In even other embodiments, the DSPE-PEG2000 is present at about 0.25 wt% to about 1 wt% of the total weight of the dry powder composition.

[0017] In one embodiment, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in about 1 wt% to about 2 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in about 0.1 wt% to about 2 wt% of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In other embodiments, the DSPE-PEG2000 is present at about 0.3 wt% to about 1.4 wt% of the total weight of the dry powder composition. In even other embodiments, the DSPE-PEG2000 is present at about 0.5 wt% to about 1 wt% of the total weight of the dry powder composition.

[0018] In one embodiment, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.2 wt% to about 1.8 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.12 wt% to about 1.8 wt% of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In other embodiments, the DSPE-PEG2000 is present at about 0.36 wt% to about 1.26 wt% of the total weight of the dry powder composition. In even other embodiments, the DSPE-PEG2000 is present at about 0.6 wt% to about 0.9 wt% of the total weight of the dry powder composition.

[0019] In one embodiment, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in about 1 wt% to about 1.5 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in about 0.1 wt% to about 1.5 wt% of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In other embodiments, the DSPE-PEG2000 is present at about 0.3 wt% to about 1.05 wt% of the total weight of the dry powder composition. In even other embodiments, the DSPE-PEG2000 is present at about 0.5 wt% to about 0.75 wt% of the total weight of the dry powder composition.

[0020] In one embodiment, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.4 wt% to about 1.6 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.14 wt% to about 1.6 wt% of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In other embodiments, the DSPE-PEG2000 is present at about 0.42 wt% to about 1.12 wt% of the total weight of the dry powder composition. In even other embodiments, the DSPE-PEG2000 is present at about 0.7 wt% to about 0.8 wt% of the total weight of the dry powder composition.

[0021] In one embodiment, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.5 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.15 wt% to about 1.5 wt% of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In other embodiments, the DSPE-PEG2000 is present at about 0.45 wt% to about 1.05 wt% of the total weight of the dry powder composition. In even other embodiments, the DSPE-PEG2000 is present at about 0.75 wt% of the total weight of the dry powder composition.

[0022] In one embodiment, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in about 0.1 wt% to about 3 wt% of the total weight of the dry powder composition, and the weight ratio of the DSPE-PEG2000 to the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is in the range of about 0.1:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt) to about 1:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt). In other embodiments, R 1 In even other embodiments, R 1 It is a linear hexadecyl group. In other embodiments, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in about 0.5wt% to about 2wt% of the total weight of the dry powder composition. In other embodiments, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in about 1wt% to about 2wt% of the total weight of the dry powder composition. In other embodiments, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in about 1.2wt% to about 1.8wt% of the total weight of the dry powder composition. In other embodiments, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in about 1wt% to about 1.5wt% of the total weight of the dry powder composition. In other embodiments, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.4wt% to about 1.6wt% of the total weight of the dry powder composition. In even other embodiments, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.5wt% of the total weight of the dry powder composition. In another embodiment, the weight ratio of the DSPE-PEG2000 to the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is in the range of about 0.3:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt) to about 0.7:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt). In other embodiments, the weight ratio of DSPE-PEG2000 to the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is about 0.5:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt).

[0023] In one embodiment of the dry powder composition provided herein, the leucine is present in about 15 wt % to about 40 wt % of the total weight of the dry powder composition. 1 In even other embodiments, R 1 is a linear hexadecyl group. In another embodiment, the leucine is present in about 18 wt % to about 33 wt % of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In another embodiment, the leucine is present in about 20 wt % to about 30 wt % of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In another embodiment, the leucine is present in about 25 wt % to about 30 wt % of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In another embodiment, the leucine is present in about 27 wt % to about 30 wt % of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 is a linear hexadecyl group. In another embodiment, the leucine is present in about 30 wt % of the total weight of the dry powder composition. In other embodiments, R 1 In even other embodiments, R 1 It is a linear hexadecyl group.

[0024] In some embodiments, the sugar is mannitol. In other embodiments, R 1 In other embodiments, R 1 It is a linear hexadecyl group.

[0025] In one embodiment, the dry powder composition comprises (a) about 1.5 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 29.3 wt% of the leucine, and the remainder (d) mannitol. In other embodiments, R 1 In other embodiments, R 1 It is a linear hexadecyl group.

[0026] In one embodiment, the dry powder composition comprises (a) about 1.5 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.75 wt% of the DSPE-PEG2000, (c) about 29.30 wt% of the leucine, and (d) about 68.45 wt% of the mannitol. In other embodiments, R 1 In other embodiments, R 1 It is a linear hexadecyl group.

[0027] In one embodiment, the dry powder composition is in the form of an aerosol having particles with a mass median aerodynamic diameter (MMAD) of about 1 μm to about 3 μm as measured by a Next Generation Impactor (NGI). In other embodiments, the dry powder composition is in the form of an aerosol having particles with a MMAD of about 1.3 μm to about 2.0 μm as measured by NGI. In other embodiments, R 1 In other embodiments, R 1 It is a linear hexadecyl group.

[0028] In one embodiment, the sugar is mannitol and the dry powder composition is in the form of an aerosol having particles with an MMAD of about 1 μm to about 3 μm as measured by NGI. In another embodiment, the sugar is mannitol and the dry powder composition is in the form of an aerosol having particles with an MMAD of about 1.7 μm to about 2.7 μm as measured by NGI. In other embodiments, R 1 In other embodiments, R 1 It is a linear hexadecyl group.

[0029] In one embodiment, the dry powder composition is in the form of an aerosol having a fine particle fraction (FPF) of about 30% to about 60% particles as measured by NGI. 1 In other embodiments, R 1 It is a linear hexadecyl group.

[0030] In another aspect, the present disclosure relates to a method for treating pulmonary hypertension in a patient in need thereof. The method comprises administering a therapeutically effective amount of a dry powder composition disclosed herein to the lungs of the patient by inhalation via a dry powder inhaler.

[0031] In one embodiment, the pulmonary hypertension is pulmonary arterial hypertension (PAH). In one embodiment, the PAH is Class I PAH as characterized by the New York Heart Association (NYHA). In another embodiment, the PAH is Class II PAH as characterized by NYHA. In another embodiment, the PAH is Class III PAH as characterized by NYHA. In another embodiment, the PAH is Class IV PAH as characterized by NYHA.

[0032] In one embodiment, the pulmonary hypertension is Group 1 pulmonary hypertension as characterized by the World Health Organization (WHO).

[0033] In another embodiment, the pulmonary hypertension is Group 2 pulmonary hypertension as characterized by the WHO.

[0034] In another embodiment, the pulmonary hypertension is Group 3 pulmonary hypertension as characterized by WHO.

[0035] In another embodiment, the pulmonary hypertension is Group 4 pulmonary hypertension as characterized by WHO.

[0036] In another embodiment, the pulmonary hypertension is Group 5 pulmonary hypertension as characterized by WHO.

[0037] In yet another aspect, the present disclosure relates to a method for treating portopulmonary hypertension or pulmonary fibrosis in a patient in need thereof. The method comprises administering a therapeutically effective amount of a dry powder composition disclosed herein to the lungs of the patient by inhalation via a dry powder inhaler.

[0038] In one embodiment of the methods of treatment described herein, the administering is in the form of once a day, twice a day, or three times a day dosing.

[0039] In another embodiment of the methods of treatment described herein, the administration comprises aerosolizing the dry powder composition and administering the aerosolized dry powder composition to the patient's lungs via inhalation. In one embodiment, the aerosolized dry powder composition comprises particles having an MMAD of about 1 μm to about 3 μm as measured by NGI. In another embodiment, the aerosolized dry powder composition comprises particles having an FPF of about 30% to about 60% as measured by NGI.

[0040] In another aspect, the present disclosure relates to a system for treating pulmonary hypertension, portopulmonary hypertension or pulmonary fibrosis. The system comprises one of the dry powder compositions disclosed herein and a dry powder inhaler (DPI).

[0041] In one embodiment, the DPI is a single-dose or multi-dose inhaler.

[0042] In another embodiment, the DPI is pre-metered or device-metered. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 is a graph showing the effect of leucine content on the spray drying recovery of mannitol-based C16TR (treprostinil palmitil) dry powder.

[0044] Figure 2A-2D are SEM images of mannitol-based C16TR (Treprostinil palmitate) dry powders containing different amounts of leucine as indicated.

[0045] Figure 3 is a graph showing the effect of leucine content on particle size distribution in mannitol-based C16TR (Treprostinil palmitate) dry powder measured by laser diffraction.

[0046] Figure 4 is a graph showing the effect of C16TR (Treprostinil palmitate) content on the MMAD of mannitol-based C16TR (Treprostinil palmitate) dry powder.

[0047] Figure 5A-5C are SEM images of mannitol-based C16TR (Treprostinil palmitate) dry powder spray dried at different inlet temperatures. The images of the upper panels were taken at high magnification, and the images of the lower panels were taken at low magnification.

[0048] Fig. 6A and Figure 6B : are SEM images showing the morphology of mannitol-based C16TR (treprostinil palmitate) dry powder spray dried at an inlet temperature of 135° C. with or without ammonium bicarbonate (ABC, 0.5 mg / mL). The images of the upper panels were taken at high magnification, and the images of the lower panels were taken at low magnification.

[0049] Fig. 7A is a graph showing DSC data for mannitol based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-179.

[0050] Figure 7B is a graph showing X-ray diffraction data for mannitol based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-179.

[0051] Fig. 8A and Figure 8B are SEM images showing the effect of leucine content on the morphology of trehalose-based C16TR (Treprostinil palmitate) dry powder.

[0052] Figure 9A-9C are SEM images showing the effect of spray drying inlet temperature on the morphology of trehalose based C16TR (Treprostinil palmitate) dry powder.

[0053] Fig. 10A is a graph showing DSC data of C16TR (Treprostinil palmitate) dry powder based on trehalose.

[0054] Fig. 10B is a graph showing X-ray diffraction data of trehalose-based C16TR (Treprostinil palmitate) dry powder.

[0055] Fig.11 is a DVS isotherm plot showing water uptake of mannitol-based C16TR (treprostinil palmitate) dry powder batch SD-NNP-167 (C16TR (treprostinil palmitate) / DSPE-PEG2000 / mannitol / leucine, 1 / 0.5 / 80 / 20).

[0056] Fig.12 is a DVS isotherm plot showing water uptake of trehalose based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-162 (C16TR (Treprostinil palmitate) / DSPE-PEG2000 / Treh / Leu, 1 / 0.5 / 80 / 20).

[0057] Fig.13 is a DVS isotherm graph showing water uptake of trehalose based C16TR (treprostinil palmitate) dry powder batch SD-NNP-163 (C16TR (treprostinil palmitate) / DSPE-PEG2000 / trehalose / leucine, 1 / 0.5 / 70 / 30).

[0058] Fig.14 is a graph showing the change in MMAD in an accelerated stability study of mannitol-based C16TR (treprostinil palmitate) dry powder.

[0059] Fig.15A and Fig. 15B are SEM images of mannitol-based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-179 containing 1% C16TR (Treprostinil palmitate) at T0 and T3 (3 months), respectively.

[0060] Fig.16A and Fig. 16Bare SEM images of mannitol-based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-183 containing 1.5% C16TR (Treprostinil palmitate) at T0 and T3 (3 months), respectively.

[0061] Fig.17A and Fig. 17B are SEM images of mannitol-based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-184 containing 2% C16TR (Treprostinil palmitate) at T0 and T3 (3 months), respectively.

[0062] Fig.18A and Fig.18B are SEM images of mannitol-based C16TR (Treprostinil palmitate) dry powder batches containing 3% C16TR (Treprostinil palmitate) (SD-NNP-190) and 5% C16TR (Treprostinil palmitate) (SD-NNP-191) at T5 (5 months), respectively.

[0063] Fig.19 is a graph showing the change in MMAD in an accelerated stability study of trehalose based C16TR (treprostinil palmitate) dry powder.

[0064] Fig. 20 is a graph showing the change in FPF in an accelerated stability study of trehalose based C16TR (treprostinil palmitate) dry powder.

[0065] Fig.21A and Fig. 21B are SEM images of trehalose-based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-162 containing 1% C16TR (Treprostinil palmitate) at T0 and T3.5 (3.5 months), respectively.

[0066] Fig.22A and Fig. 22B are SEM images of trehalose-based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-163 containing 1% C16TR (Treprostinil palmitate) at T0 and T3.5 (3.5 months), respectively.

[0067] Fig.23A and Fig. 23B are SEM images of trehalose-based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-188 containing 1.5% C16TR (Treprostinil palmitate) at T0 and T3.5 (3.5 months), respectively.

[0068] Fig.24A and Fig. 24Bare SEM images of trehalose-based C16TR (Treprostinil palmitate) dry powder batch SD-NNP-189 containing 2% C16TR (Treprostinil palmitate) at T0 and T3.5 (3.5 months), respectively.

[0069] Fig.25 is a graph showing pressure titrations of spray-dried Treprostinil palmitate dry powder formulations A, B, C, and D. For visibility, the data points are offset (left to right) in order (A, B, C, and D) within each gas pressure category.

[0070] Fig.26 is a graph showing the particle size distribution of Treprostinil palmitate dry powder formulations A, B, C and D.

[0071] Fig. 27 is a SEM image of Treprostinil palmitate dry powder Formulation A.

[0072] Fig.28 is a SEM image of Treprostinil palmitate dry powder Formulation B.

[0073] Fig.29 is a SEM image of Treprostinil palmitate dry powder Formulation C.

[0074] Fig.30 is a SEM image of Treprostinil palmitate dry powder Formulation D.

[0075] Fig.31 is a graph showing the fine particle dose (FPD) of Treprostinil palmitate dry powder Formulation A at T=0 and after storage in capsules for 1-3 months at 25°C or 40°C as indicated or after storage in bulk for 3 months at 25°C or 40°C and filled into capsules and administered on the same day.

[0076] Fig.32 is a graph showing the fine particle dose (FPD) of Treprostinil palmitate dry powder Formulation C at T=0 and after storage in capsules for 1-3 months at 25°C or 40°C as indicated or after storage in bulk for 3 months at 25°C or 40°C and filled into capsules and administered on the same day.

[0077] Fig.33 is a graph showing the fine particle dose (FPD) of Treprostinil palmitate dry powder Formulation D at T=0 and after storage in capsules for 1-3 months at 25°C or 40°C as indicated or after storage in bulk for 3 months at 25°C or 40°C and filled into capsules and administered on the same day.

[0078] Fig.34 is a dynamic vapor sorption (DVS) isotherm plot of trehalose based C16TR (treprostinil palmitate) dry powder formulation.

[0079] Fig.35 is a graph showing the aerosol particle size distribution of trehalose-based C16TR (treprostinil palmitate) dry powder formulation at T=0 and after storage at 40°C and uncontrolled ambient humidity for 1.5 months, 2.5 months and 3.5 months (each time point, n=1).

[0080] Fig.36 is a graph showing the concentration of C16TR (Treprostinil palmitate) equivalents (C16TR (Treprostinil palmitate) plus Treprostinil, ng / g) in the lung after inhalation of a trehalose-based C16TR (Treprostinil palmitate) dry powder formulation and aerosolized INS1009.

[0081] Fig.37 is a graph showing the concentration of C16TR (treprostinil palmitate) equivalent (C16TReq) in the lung after inhalation of Treprostinil palmitate dry powder Formulation D or Formulation C.

[0082] Fig.38 is a graph showing the concentration of C16TR (Treprostinil palmitate) in the lung after inhalation of Treprostinil palmitate dry powder Formulation D or Formulation C.

[0083] Fig.39 is a graph showing the concentration of TRE in the lung after inhalation of Treprostinil palmitate dry powder Formulation D or Formulation C.

[0084] Fig.40 is a graph showing the concentration of TRE in plasma after inhalation of Treprostinil palmitate dry powder Formulation D or Formulation C.

[0085] Fig.41 is a graph showing the ΔRVPP response to hypoxic challenge in rats exposed to Treprostinil palmitate dry powder Formulation D or Formulation C. DETAILED DESCRIPTION

[0086] Throughout this disclosure, the term "about" can be used in conjunction with numerical values ​​and / or ranges. The term "about" is understood to mean those values ​​close to the value. For example, "about 40 [units]" can mean ± 25% (e.g., from 30 to 50) of 40, such as ± 20%, ± 15%, ± 10%, ± 9%, ± 8%, ± 7%, ± 6%, ± 5%, ± 4%, ± 3%, ± 2%, ± 1%, less than ± 1% of 40, or other values ​​or ranges therebetween or lower values.

[0087] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid (including an inorganic or organic base and an inorganic or organic acid). The nature of the salt is not critical, provided that it is pharmaceutically acceptable. Suitable pharmaceutically acceptable acid addition salts can be prepared from inorganic acids or from organic acids. Exemplary pharmaceutical salts are disclosed in Stahl, PH, Wermuth, CG, ed., Handbook of Pharmaceutical Salts: Properties, Selection and Use; Verlag Helvetica Chimica Acta / Wiley-VCH: Zurich, 2002, the contents of which are incorporated herein by reference in their entirety. Specific non-limiting examples of inorganic acids are hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, and phosphoric acid. Suitable organic acids include, but are not limited to, fatty acids, alicyclic acids, aromatic acids, aryl fatty acids, and carboxylic acids and sulfonic acids containing heterocyclic groups, such as formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, methanesulfonic acid, stearic acid, salicylic acid, p-hydroxybenzoic acid, phenylacetic acid, mandelic acid, pamoic acid (pamoic acid), methanesulfonic acid, ethylsulfonic acid, benzenesulfonic acid, pantothenic acid, toluenesulfonic acid, 2-hydroxyethylsulfonic acid, sulfanilic acid, cyclohexylaminosulfonic acid, alginic acid, 3-hydroxybutyric acid, galactonic acid or galacturonic acid. Suitable pharmaceutically acceptable salts of compounds containing free acids disclosed herein include, but are not limited to, metal salts and organic salts. Exemplary metal salts include, but are not limited to, suitable alkali metal (Ia group) salts, alkaline earth metal (IIa group) salts and other physiologically acceptable metals. Such salts can be made from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc. Exemplary organic salts can be made from primary, secondary, tertiary, and quaternary ammonium salts, such as tromethamine, diethylamine, tetra-N-methylammonium, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine.

[0088] Throughout this specification, numerical ranges are provided for certain quantities. It should be understood that these ranges include all subranges therein. Therefore, the range of "50-80" includes all possible ranges therein (e.g., 51-79, 52-78, 53-77, 54-76, 55-75, 60-70, etc.). In addition, all values ​​within a given range can be endpoints of the ranges thus covered (e.g., the range 50--80 includes ranges with endpoints, such as 55-80, 50-75, etc.).

[0089] In one embodiment, the term "treating" includes: (1) preventing or delaying the onset of clinical symptoms of a condition, disorder, or condition that develops in a subject who may have or be predisposed to the condition, disorder, or condition but has not yet experienced or displayed clinical or subclinical symptoms of the condition, disorder, or condition; (2) inhibiting the condition, disorder, or condition (e.g., preventing, alleviating, or delaying the development of the disease, or the recurrence of at least one clinical or subclinical symptom thereof in the case of maintenance therapy); and / or (3) alleviating the condition (e.g., causing regression of the condition, disorder, or condition, or at least one clinical or subclinical symptom thereof). In one embodiment, "treating" refers to inhibiting the condition, disorder, or condition (e.g., preventing, alleviating, or delaying the development of the disease, or the recurrence of at least one clinical or subclinical symptom thereof in the case of maintenance therapy). In another embodiment, "treating" refers to alleviating the condition (e.g., alleviating by causing regression of at least one of the condition, disorder, or condition, or its clinical or subclinical symptoms). The benefit to the treated subject is statistically significant, or at least perceptible to the subject or physician, as compared to the same subject's state or condition prior to such treatment, or as compared to the state or condition of an untreated control subject.

[0090] "Effective amount" refers to an amount of a dry powder composition of the present disclosure sufficient to result in a desired therapeutic response.

[0091] In one aspect of the present invention, a dry powder composition of a Treprostinil prodrug is provided. The dry powder composition comprises: (a) Compounds of formula (I): or an enantiomer, a diastereomer or a pharmaceutically acceptable salt thereof, wherein R 1 is tetradecyl, pentadecyl, hexadecyl, heptadecyl or octadecyl, and the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof is present in an amount of about 0.1 wt % to about 3 wt % of the total weight of the dry powder composition; (b) about 0.01 wt % to about 3 wt % DSPE-PEG2000; (c) from about 10 wt % to about 50 wt % of leucine; and the remainder is (d) A sugar selected from trehalose and mannitol. The total of (a), (b), (c) and (d) is 100 wt%.

[0092] In some embodiments, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 0.1 wt%, about 0.3 wt%, about 0.5 wt%, about 0.7 wt%, about 1 wt%, about 1.3 wt%, about 1.5 wt%, about 1.7 wt%, about 2.0 wt%, about 2.3 wt%, about 2.5 wt%, about 2.7 wt% or about 3 wt% of the total weight of the dry powder composition. In other embodiments, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.5 wt% of the total weight of the dry powder composition. The compound of formula (I) and its pharmaceutically acceptable salt are treprostinil prodrugs disclosed in International Application Publication WO 2015 / 061720, the disclosure of which is incorporated herein by reference in its entirety. In some embodiments, the leucine is present at about 10 wt%, about 15 wt%, about 20 wt%, about 25 wt%, about 30 wt%, about 35 wt%, about 40 wt%, about 45 wt%, or about 50 wt% of the total weight of the dry powder composition.

[0093] PEG refers to polyethylene glycol, also known as polyethylene oxide (PEO) or polyoxyethylene (POE), depending on its molecular weight. The DSPE-PEG2000 can include branched or unbranched PEG molecules with an average PEG molecular weight of 2000 g / mol. In one embodiment, (b) is DSPE-PEG2000 present in an amount of about 0.03 wt% to about 2.1 wt% of the gross weight of the dry powder composition. In another embodiment, (b) is DSPE-PEG2000 present in an amount of about 0.05 wt% to about 1.5 wt% of the gross weight of the dry powder composition.

[0094] In one embodiment of the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, R 1 In other embodiments, R 1 It is a linear tetradecyl group.

[0095] In another embodiment of the compound of Formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, R 1 In other embodiments, R 1 It is a linear pentadecyl group.

[0096] In another embodiment of the compound of Formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, R 1 In other embodiments, R 1 It is a linear heptadecanyl group.

[0097] In another embodiment of the compound of Formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, R 1 In other embodiments, R 1 It is a linear octadecyl group.

[0098] In another embodiment of the compound of Formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, R 1 In other embodiments, R 1 is a linear hexadecyl group, that is, the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is a compound of formula (II): Or an enantiomer, a diastereomer, or a pharmaceutically acceptable salt thereof. In one embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is a compound of formula (II) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof is a compound of formula (II). In other embodiments, the compound of formula (I) is a compound of formula (II). The compound of formula (II) is also referred to herein as C16TR or Treprostinil palmitate. In the present application, C16TR and Treprostinil palmitate are used interchangeably and refer to the compound of formula (II).

[0099] In one embodiment, (a) is a compound of formula (I) or a pharmaceutically acceptable salt thereof. In other embodiments, (a) is a compound of formula (I). In another embodiment, (a) is a compound of formula (II) or a pharmaceutically acceptable salt thereof. In other embodiments, (a) is a compound of formula (II).

[0100] In one embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 0.5wt% to about 2wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.05wt% to about 2wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.15wt% to about 1.4wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.25wt% to about 1wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) or its pharmaceutically acceptable salt is present at about 0.5wt% to about 2wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) is present at about 0.5wt% to about 2wt% of the total weight of the dry powder composition.

[0101] In one embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in about 1wt% to about 2wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in about 0.1wt% to about 2wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present in about 0.3wt% to about 1.4wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present in about 0.5wt% to about 1wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) or its pharmaceutically acceptable salt is present in about 1wt% to about 2wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) is present in about 1wt% to about 2wt% of the total weight of the dry powder composition.

[0102] In one embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.2wt% to about 1.8wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.12wt% to about 1.8wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.36wt% to about 1.26wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.6wt% to about 0.9wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) or its pharmaceutically acceptable salt is present at about 1.2wt% to about 1.8wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) is present at about 1.2wt% to about 1.8wt% of the total weight of the dry powder composition.

[0103] In one embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1wt% to about 1.5wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.1wt% to about 1.5wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.3wt% to about 1.05wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.5wt% to about 0.75wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) or its pharmaceutically acceptable salt is present at about 1wt% to about 1.5wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) is present at about 1wt% to about 1.5wt% of the total weight of the dry powder composition.

[0104] In one embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.4wt% to about 1.6wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.14wt% to about 1.6wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.42wt% to about 1.12wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.7wt% to about 0.8wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) or its pharmaceutically acceptable salt is present at about 1.4wt% to about 1.6wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) is present at about 1.4wt% to about 1.6wt% of the total weight of the dry powder composition.

[0105] In one embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.1wt% to about 1wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.3wt% to about 0.7wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.5wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) or its pharmaceutically acceptable salt is present at about 1wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) is present at about 1wt% of the total weight of the dry powder composition.

[0106] In one embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.5wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.15wt% to about 1.5wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.45wt% to about 1.05wt% of the total weight of the dry powder composition. In other embodiments, the DSPE-PEG2000 is present at about 0.75wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) or its pharmaceutically acceptable salt is present at about 1.5wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) is present at about 1.5wt% of the total weight of the dry powder composition.

[0107] In some embodiments, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 0.1 wt% to about 3 wt%, about 0.5 wt% to about 2 wt%, about 1 wt% to about 2 wt%, about 1.2 wt% to about 1.8 wt%, about 1 wt% to about 1.5 wt%, about 1.4 wt% to about 1.6 wt%, about 1 wt%, or about 1.5 wt% of the total weight of the dry powder composition, and the weight of the DSPE-PEG2000 and the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is about 1.5 wt%. The ratio is in the range of about 0.1:1 (DSPE-PEG2000: compound of formula (I) or (II)) to about 1:1 (DSPE-PEG2000: compound of formula (I) or (II)) or about 0.3:1 (DSPE-PEG2000: compound of formula (I) or (II)) to about 0.7:1 (DSPE-PEG2000: compound of formula (I) or (II)), for example, about 0.1:1, about 0.2:1, about 0.3:1, about 0.4:1, about 0.5:1, about 0.6:1, about 0.7:1, about 0.8:1, about 0.9:1 or about 1:1. In one embodiment, the weight ratio of DSPE-PEG2000 to the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is in the range of about 0.1:1 (DSPE-PEG2000: compound of formula (I) or (II)) to about 1:1 (DSPE-PEG2000: compound of formula (I) or (II)). In another embodiment, the weight ratio of DSPE-PEG2000 to the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is in the range of about 0.3:1 (DSPE-PEG2000: compound of formula (I) or (II)) to about 0.7:1 (DSPE-PEG2000: compound of formula (I) or (II)).

[0108] In some embodiments, the weight ratio of the DSPE-PEG2000 to the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is about 0.5:1 (DSPE-PEG2000: compound of formula (I) or (II)). At this weight ratio, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1wt% to about 2wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.5wt% to about 1wt% of the total weight of the dry powder composition. In another embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.2wt% to about 1.8wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.6wt% to about 0.9wt% of the total weight of the dry powder composition. In another embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.4wt% to about 1.6wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.7wt% to about 0.8wt% of the total weight of the dry powder composition. In another embodiment, the compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.5wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.75wt% of the total weight of the dry powder composition. In some embodiments, the compound of formula (I) or (II) or its pharmaceutically acceptable salt is present in each of the above-mentioned weight percentages or weight percentage ranges in the dry powder composition. In some embodiments, the compound of formula (I) or (II) is present in each of the above-mentioned weight percentages or weight percentage ranges in the dry powder composition.

[0109] In one embodiment, the leucine is present at about 15wt% to about 40wt% of the total weight of the dry powder composition. In other embodiments, the leucine is present at about 18wt% to about 33wt% of the total weight of the dry powder composition. In other embodiments, the leucine is present at about 20wt% to about 30wt%, such as about 20wt%, about 25wt% or about 30wt% of the total weight of the dry powder composition. In other embodiments, the leucine is present at about 25wt% to about 30wt% of the total weight of the dry powder composition. In other embodiments, the leucine is present at about 27wt% to about 30wt% of the total weight of the dry powder composition. In one embodiment, the leucine is present at about 20wt% of the total weight of the dry powder composition. In another embodiment, the leucine is present at about 30wt% of the total weight of the dry powder composition.

[0110] In some embodiments, the sugar in the dry powder composition is trehalose. In other embodiments, the sugar in the dry powder composition is mannitol.

[0111] In one embodiment, the dry powder composition comprises (a) about 1.5 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 29.3 wt% of the leucine, and the remainder (d) trehalose. In other embodiments, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II). In some embodiments, R 1 is hexadecyl in the compound of formula (I). In other embodiments, R 1 is the linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is the compound of formula (II).

[0112] In another embodiment, the dry powder composition comprises (a) about 1 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 29.6 wt% of the leucine, and the remainder (d) trehalose. In other embodiments, (a) in the dry powder composition is about 1 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1 wt% of a compound of formula (I) or (II). In some embodiments, R 1is hexadecyl in the compound of formula (I). In other embodiments, R 1 is the linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is the compound of formula (II).

[0113] In another embodiment, the dry powder composition comprises (a) about 1 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 19.7 wt% of the leucine, and the remainder (d) trehalose. In other embodiments, (a) in the dry powder composition is about 1 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1 wt% of a compound of formula (I) or (II). In some embodiments, R 1 is hexadecyl in the compound of formula (I). In other embodiments, R 1 is the linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is the compound of formula (II).

[0114] In another embodiment, the dry powder composition comprises (a) about 1.5 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 19.6 wt% of the leucine, and the remainder (d) trehalose. In other embodiments, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II). In some embodiments, R 1 is hexadecyl in the compound of formula (I). In other embodiments, R 1 is the linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is the compound of formula (II).

[0115] In another embodiment, the dry powder composition comprises (a) about 1.5 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 29.3 wt% of the leucine, and the remainder (d) mannitol. In other embodiments, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II). In some embodiments, R 1 is hexadecyl in the compound of formula (I). In other embodiments, R 1 is the linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is the compound of formula (II).

[0116] In another embodiment, the dry powder composition comprises (a) about 1.5 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.75 wt% of the DSPE-PEG2000, (c) about 29.30 wt% of the leucine, and (d) about 68.45 wt% of the mannitol. In other embodiments, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II). In some embodiments, R 1 is hexadecyl in the compound of formula (I). In other embodiments, R 1 is a linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is a compound of formula (II). In one embodiment, the dry powder composition comprises (a) about 1.5wt% of the compound of formula (II), (b) about 0.75wt% of the DSPE-PEG2000, (c) about 29.30wt% of the leucine, and (d) about 68.45wt% of the mannitol.

[0117] In another embodiment, the dry powder composition comprises (a) about 1 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 29.6 wt% of the leucine, and the remainder (d) mannitol. In other embodiments, (a) in the dry powder composition is about 1 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1 wt% of a compound of formula (I) or (II). In some embodiments, R 1 is hexadecyl in the compound of formula (I). In other embodiments, R 1 is the linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is the compound of formula (II).

[0118] In another embodiment, the dry powder composition comprises (a) about 1.5 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 19.6 wt% of the leucine, and the remainder (d) mannitol. In other embodiments, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1.5 wt% of a compound of formula (I) or (II). In some embodiments, R 1 is hexadecyl in the compound of formula (I). In other embodiments, R 1 is the linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is the compound of formula (II).

[0119] In another embodiment, the dry powder composition comprises (a) about 1 wt% of a compound of formula (I) or (II) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 19.7 wt% of the leucine, and the remainder (d) mannitol. In other embodiments, (a) in the dry powder composition is about 1 wt% of a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, (a) in the dry powder composition is about 1 wt% of a compound of formula (I) or (II). In some embodiments, R 1 is hexadecyl in the compound of formula (I). In other embodiments, R 1is the linear hexadecyl group in the compound of formula (I), that is, the compound of formula (I) is the compound of formula (II).

[0120] Mass median aerodynamic diameter (MMAD) is the value of the following aerodynamic diameter, wherein 50% of the mass in a given aerosol is associated with particles less than the median aerodynamic diameter (MAD), and 50% of the mass is associated with particles greater than MAD. MMAD can be determined by impactor measurements, such as Anderson cascade impactor (ACT) or next generation impactor (NGI). In some embodiments, the dry powder composition is in the form of an aerosol, comprising particles having MMAD of 1 μm to about 5 μm, about 1 μm to about 3 μm, about 1.3 μm to about 2.0 μm, or about 1.7 μm to about 2.7 μm as measured by NGI. In one embodiment, the sugar in the dry powder composition is mannitol. In another embodiment, the sugar in the dry powder composition is trehalose.

[0121] In one embodiment, the sugar in the dry powder composition is mannitol, and the dry powder composition is in the form of an aerosol comprising particles having an MMAD of about 1 μm to about 3 μm as measured by NGI. In another embodiment, the sugar in the dry powder composition is mannitol, and the dry powder composition is in the form of an aerosol comprising particles having an MMAD of about 1.7 μm to about 2.7 μm as measured by NGI.

[0122] "Fine particle fraction" or "FPF" refers to the fraction of aerosols having a particle size less than 5 μm as measured by cascade impaction. FPF is usually expressed as a percentage. FPF has been shown to be associated with the fraction of powder deposited in the lungs of patients. In some embodiments, the dry powder composition is in the form of an aerosol comprising particles having a FPF of at least 20%, at least 30%, at least 40%, at least 50%, about 30% to about 60%, about 35% to about 55%, or about 40% to about 50% as measured by NGI. In one embodiment, the sugar in the dry powder composition is mannitol. In another embodiment, the sugar is trehalose.

[0123] The tap density of a powder is the ratio of the mass of the powder to the volume occupied by the powder after tapping within a defined time period. The tap density of a powder represents its random dense packing. The tap density can be measured using the method of USP Bulk Density and Tapped Density, United States Pharmacopeia convention, Rockville, Md., 10th Supplement, 4950-4951, 1999. In some embodiments, the dry powder composition comprises particles having a tap density of about 0.2 g / ml to about 0.8 g / ml or about 0.3 g / ml to about 0.6 g / ml. In one embodiment, the sugar in the dry powder composition is mannitol. In another embodiment, the sugar in the dry powder composition is trehalose.

[0124] The dry powder compositions of the present disclosure can be produced from liquid compositions using freeze-drying or spray-drying techniques. When freeze-drying is used, the freeze-dried composition can be ground to obtain a finely divided dry powder containing particles within the above-mentioned desired size range. When spray-drying is used, the method is carried out under conditions that can produce a finely divided dry powder containing particles within the above-mentioned desired size range. Exemplary methods for preparing pharmaceutical compositions in dry powder form are disclosed in WO 96 / 32149, WO 97 / 41833, WO 98 / 29096, and U.S. Pat. Nos. 5,976,574, 5,985,248, and 6,001,336; the disclosures of each of which are incorporated herein by reference in their entirety. Exemplary spray-drying methods are described in U.S. Pat. Nos. 6,848,197 and 8,197,845, the disclosures of each of which are incorporated herein by reference in their entirety.

[0125] In some embodiments, the dry powder composition of the present disclosure is prepared by the following process. Use organic solvents such as alcohol (e.g., 1-propanol) to prepare a stock solution of a compound of formula (I) or (II) or its enantiomer, diastereomer or pharmaceutically acceptable salt and the DSPE-PEG2000. Also prepare an aqueous stock solution of sugar (e.g., mannitol or trehalose) and leucine. Then the required amount of the above stock solution is added to a mixture of water and an organic solvent to form a spray-dried feed solution. In the spray-dried feed solution, the volume ratio of water to the organic solvent can be about 3:2 to about 1:1.

[0126] Spray drying is started in the following manner: the drying gas flow is turned on and the drying gas is heated by setting the required inlet temperature to, for example, about 120°C to about 160°C or about 135°C to about 150°C. After the spray dryer outlet temperature reaches a suitable temperature (e.g., about 55°C to about 65°C), a liquid slide inlet is set to allow the blank solvent to be atomized into the spray dryer by nitrogen, and the system is cooled and stabilized. The product filter pulse is started and the product filter purge flow is set to, for example, 10 to 20 scfh. After the system is stabilized, the liquid slide inlet is switched to the feed solution prepared above, and the process is continued until the feed solution is exhausted. At the time point when the feed solution is exhausted, the liquid slide inlet is switched back to the blank solvent, which is sprayed for about 5 to about 20 minutes. At this point, the powder is collected at the bottom of the product filter. After the blank solvent is sprayed for about 5 to about 20 minutes, the system is closed by closing the liquid pipeline, the atomizing gas, the drying gas heater, the drying gas inlet, and the final exhaust pipe.

[0127] In one embodiment, the dry powder composition of the present disclosure is delivered to the lungs of the experimenter via suction using a dry powder inhaler (DPI). In one embodiment, the dry powder inhaler is a single-dose dry powder inhaler. DPI, as a propellant-free device, utilizes the experimenter to inhale and deliver the dry powder to the lungs of the experimenter. The unit dose of the dry powder composition used in the DPI device is usually the dry powder blister disk of a hard capsule. The exemplary DPI device suitable for delivering the dry powder composition of the present disclosure includes the device described in the following paragraphs and the DPI described in U.S. Patent Nos. 6,766,799, 7,278,425 and 8,496,002, and the disclosure of each of them is incorporated herein by reference in its entirety.

[0128] The inhaler (Alkermes) includes a small breath-activated system that delivers a porous powder from a capsule. The porous particles have an aerodynamic diameter of 1-5 μm. See International Patent Application Publication Nos. WO 99 / 66903 and WO 00 / 10541, the disclosures of each of which are incorporated herein by reference in their entirety.

[0129] Aerolizer TM (Novartis) is a single-dose dry powder inhaler. In this device, the dry powder drug is stored in a capsule and released by piercing the capsule wall with a TEFLON-coated steel needle. See U.S. Patent Nos. 6,488,027 and 3,991,761, the disclosures of each of which are incorporated herein by reference in their entirety.

[0130] Bang Olufsen provides a breath-actuated inhaler using a blister strip of up to 60 doses. The dose is delivered only during inhalation by a novel trigger mechanism. The device is equipped with a dose counter and can be discarded after all doses have been used. See EP 1522325, the disclosure of which is incorporated herein by reference in its entirety.

[0131] (Innovata PLC) is a large reservoir breath-activated multi-dose device. See U.S. Patent 5,437,270, the disclosure of which is incorporated herein by reference in its entirety.

[0132] DirectHaler TM (Direct-Haler A / S) is a single-dose, pre-measured, pre-filled, disposable DPI device made of polypropylene. See U.S. Patent No. 5,797,392, the disclosure of which is incorporated herein by reference in its entirety.

[0133] Diskus TM (GlaxoSmithKline) is a disposable miniature DPI device containing up to 60 doses contained in a double foil blister strip to provide moisture protection. See GB 2242134, the disclosure of which is incorporated herein by reference in its entirety.

[0134] Eclipse TM (Aventis) is a breath-actuated, reusable capsule device capable of delivering up to 20 mg of a dry powder composition. The powder is drawn from the capsule into a vortex chamber where a rotating ball helps the powder deagglomerate as the subject inhales. See U.S. Pat. No. 6,230,707 and WO 9503846, the disclosures of each of which are incorporated herein by reference in their entirety.

[0135] is a plastic breath-activated dry powder inhaler and is suitable for use with the dry powder compositions provided herein.

[0136] (Hovione) is a capsule-based, refillable, reusable passive dry powder inhaler that holds up to 14 capsules. The inhaler itself is moisture resistant. See U.S. Pat. No. 5,673,686, the disclosure of which is incorporated herein by reference in its entirety.

[0137] (Vectura) is a passive disposable DPI containing a blister strip. See GB2407042, the disclosure of which is incorporated herein by reference in its entirety.

[0138] (Boehringer Ingelheim GmbH) is a single-dose DPI device. It can provide up to 30 mg of dry powder composition in a capsule. See International Patent Application Publication No. WO 04 / 024156, the disclosure of which is incorporated herein by reference in its entirety.

[0139] MicroDose DPI (Microdose Technologies) is a small electronic DPI device. It uses a piezoelectric vibrator (ultrasonic frequency) to deagglomerate drug powder in aluminum blisters (single or multi-dose). See U.S. Patent No. 6,026,809, the disclosure of which is incorporated herein by reference in its entirety.

[0140] Nektar Dry Powder (Nektar) is a palm-sized, easy-to-use device that provides convenient dosing from a standard capsule and lung deposition independent of flow rate.

[0141] Nektar Pulmonary (Nektar) effectively removes powder from packaging, breaks down particles and forms an aerosol cloud suitable for deep lung delivery. It enables aerosolized particles to be delivered from the device to the deep lung during the patient's breathing while reducing losses in the throat and upper airway. Compressed gas is used to aerosolize the powder. See AU4090599 and U.S. Patent No. 5,740,794, the disclosure of each of which is incorporated herein by reference in its entirety.

[0142] NEXT DPI TM It is a device with multi-dose capacity, moisture protection and dose counting function. The device can be used only when the appropriate inspiratory flow is achieved, regardless of the direction (upward or downward) and the dose. See EP1196146, U.S. Patent No. 6,528,096, WO 0178693 and WO 0053158, the disclosure of each of which is incorporated herein by reference in its entirety.

[0143] is a capsule-based, plastic, breath-activated dry powder inhaler.

[0144] Oriel TM The DPI is an active DPI that utilizes a piezoelectric membrane and non-linear vibrations to aerosolize powder formulations. See International Patent Application Publication No. WO 0168169, the disclosure of which is incorporated herein by reference in its entirety.

[0145] The RS01 single-dose dry powder inhaler, developed by Plastiape of Italy, features a compact size and an effective perforation system, and is suitable for both gelatin and HMPC capsules.

[0146] Pressair TM It is a plastic breath-activated dry powder inhaler.

[0147] The Chiesi is a breath-actuated multi-dose (100 doses) dry powder inhaler. The dry powder is stored in a reservoir that is transparent and clearly marked to indicate when the 100th dose has been delivered. See U.S. Patent No. 5,351,683, the disclosure of which is incorporated herein by reference in its entirety.

[0148] (GlaxoSmithKline) is a disposable device that uses a capsule. See U.S. Patent Nos. 5,673,686 and 5,881,721, the disclosures of each of which are incorporated herein by reference in their entirety.

[0149] Rexam DPI (Rexam Pharma) is a single-dose, reusable device designed for use with capsules. See US Pat. No. 5,651,359 and EP 0707862, the disclosures of each of which are incorporated herein by reference in their entirety.

[0150] S2 (Innovata PLC) is a reusable or disposable single-dose DPI for delivering high-concentration dry powder compositions. Its dispersion mechanism requires minimal effort from the patient to achieve excellent delivery of the drug to the patient's lungs. S2 is easy to use and has a passive engine, so no batteries or power supply are required. See AU3320101, the disclosure of which is incorporated herein by reference in its entirety.

[0151] DPI (SkyePharma) is a multi-dose device that contains up to 300 individual doses in a disposable or replaceable cartridge. The device is driven by breathing and does not require coordination between breathing and actuation. See U.S. Pat. No. 6,182,655 and WO 97 / 20589, the disclosures of each of which are incorporated herein by reference in their entirety.

[0152] DPI (LAB International) is a multi-dose (up to 200) DPI device. It is breath-driven and independent of flow rate. The device includes a unique water-balanced drug reservoir coupled to a volumetric dose metering system for consistent dosing. See U.S. Patent No. 6,132,394, the disclosure of which is incorporated herein by reference in its entirety.

[0153] (AstraZeneca) is described in U.S. Pat. No. 5,983,893, the disclosure of which is incorporated herein by reference in its entirety. The DPI device is an inspiratory flow-driven, multi-dose dry powder inhaler with a multi-dose reservoir that provides up to 200 doses of a dry powder composition, and the dose range is from a few micrograms to 0.5 mg.

[0154] (Schering-Plough) is a multi-dose device with dose counting function and capable of 14-200 actuations. The dry powder composition is packaged in a cartridge containing a desiccant. See U.S. Patent No. 5,829,434, the disclosure of which is incorporated herein by reference in its entirety.

[0155] (Aventis) combines accurate dose metering with good dispersibility. It is an easy-to-use, discrete, pocket-sized device with a digital dose counter, dose taking indicator and locking mechanism. The device is capable of delivering up to 20 mg of dry powder composition. Described in US Pat. No. 5,678,538 and WO 2004026380, the disclosures of each of which are incorporated herein by reference in their entirety.

[0156] Xcelovair TM (Meridica / Pfizer) holds 60 pre-measured, airtight sealed doses ranging from 5-20 mg. The device provides moisture protection under accelerated conditions of 40°C / 75% RH. The dispersion system maximizes the fine particle fraction, delivering up to 50% fine particle mass.

[0157] On the other hand, a system is provided, the system comprising (i) one of the dry powder compositions described herein and (ii) a dry powder inhaler (DPI) for administering the dry powder composition. The DPI comprises (a) a reservoir comprising the dry powder composition disclosed herein and (b) a device for introducing the dry powder composition into the patient via inhalation. In one embodiment, the reservoir comprises the dry powder composition of the present invention in a capsule or blister package. The material of the capsule shell can be gelatin, a cellulose derivative, starch, a starch derivative, chitosan or a synthetic plastic. The DPI can be a single-dose or multi-dose inhaler. In addition, the DPI can be a pre-metered or device-metered formula. In one embodiment, the dry powder inhaler is a single-dose dry powder inhaler.

[0158] In one embodiment, the system is used to treat pulmonary hypertension, portopulmonary hypertension or pulmonary fibrosis. The system includes a dry powder composition disclosed herein, i.e., a dry powder composition comprising a compound of formula (I) or (II) or an enantiomer, diastereomer or a pharmaceutically acceptable salt thereof; and DPI. In one embodiment, the dry powder composition comprises a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, the dry powder composition comprises a compound of formula (I) or (II). The dry powder inhaler may be one of the above, may be a single-dose or multi-dose inhaler, and / or may be a pre-metered or device-metered inhaler. In one embodiment, the dry powder inhaler is a single-dose dry powder inhaler.

[0159] In another aspect of the present invention, a method for treating pulmonary hypertension (PH) in a patient in need is provided. The method includes administering an effective amount of a dry powder composition disclosed herein, i.e., a dry powder composition comprising a compound of formula (I) or (II) or its enantiomer, diastereomer, or pharmaceutically acceptable salt, to the patient's lungs by inhalation via a dry powder inhaler. In one embodiment, the dry powder composition comprises a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, the dry powder composition comprises a compound of formula (I) or (II). In one embodiment, the administration includes atomizing the dry powder composition via a DPI to provide an atomized dry powder composition, and administering the atomized dry powder composition to the patient's lungs via inhalation via the DPI. In some embodiments, the atomized dry powder composition comprises particles having an MMAD of 1 μm to about 10 μm, about 1 μm to about 7 μm, about 1 μm to about 5 μm, or about 1 μm to about 3 μm as measured by NGI. In one embodiment, the aerosolized dry powder composition comprises particles having a FPF of about 40% to about 70%, about 30% to about 60%, or about 50% to about 60% as measured by NGI.

[0160] The World Health Organization (WHO) divides PH into five groups. Group 1 PH includes pulmonary arterial hypertension (PAH), idiopathic pulmonary arterial hypertension (IPAH), familial pulmonary arterial hypertension (FPAH), and pulmonary arterial hypertension (APAH) associated with other diseases. For example, pulmonary arterial hypertension associated with collagen vascular disease (e.g., scleroderma), congenital shunts between systemic and pulmonary circulations, portal hypertension, and / or HIV infection is included in Group 1 PH. Group 2 PH includes pulmonary hypertension associated with left heart disease (e.g., atrial or ventricular disease) or valvular disease (e.g., mitral stenosis). WHO Group 3 PH is characterized by pulmonary hypertension associated with lung disease (e.g., chronic obstructive pulmonary disease (COPD), interstitial lung disease (ILD), and / or hypoxemia). Group 4 PH is pulmonary hypertension caused by chronic thrombosis and / or embolic disease. Group 4 PH is also called chronic thromboembolic pulmonary hypertension. Group 4 PH patients experience vascular obstruction or narrowing caused by blood clots. Group 5 PH is the “miscellaneous” category and includes PH caused by, for example, blood disorders (e.g., polycythemia vera, essential thrombocythemia), systemic disorders (e.g., sarcoidosis, vasculitis), and / or metabolic disorders (e.g., thyroid disease, glycogen storage disease).

[0161] The methods provided herein can be used to treat patients with PH as characterized by WHO as Group 1 (i.e., pulmonary arterial hypertension or PAH), Group 2, Group 3, Group 4, or Group 5. In one embodiment of the method, the pulmonary hypertension treated is chronic thromboembolic pulmonary hypertension.

[0162] In another embodiment of the method, the pulmonary hypertension treated is pulmonary arterial hypertension (PAH).In some embodiments, the PAH treated is Class I PAH, Class II PAH, Class III PAH, or Class IV PAH as characterized by the New York Heart Association (NYHA).

[0163] In one embodiment, the PAH is Class I PAH as characterized by NYHA.

[0164] In another embodiment, the PAH is Class II PAH as characterized by NYHA.

[0165] In yet another embodiment, the PAH is Class III PAH as characterized by NYHA.

[0166] In yet another embodiment, the PAH is Class IV PAH as characterized by NYHA.

[0167] On the other hand, the present disclosure provides a method for treating portopulmonary hypertension (PPH) in patients in need thereof. The method includes administering an effective amount of a dry powder composition disclosed herein, i.e., a dry powder composition comprising a compound of formula (I) or (II) or its enantiomer, diastereomer, or pharmaceutically acceptable salt, to the patient's lungs by inhalation via a dry powder inhaler. In one embodiment, the dry powder composition comprises a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, the dry powder composition comprises a compound of formula (I) or (II). In one embodiment, the administration includes atomizing the dry powder composition with a dry powder inhaler (DPI) to provide an atomized dry powder composition, and administering the atomized dry powder composition to the patient's lungs via the DPI. In some embodiments, the atomized dry powder composition comprises particles having an MMAD of 1 μm to about 10 μm, about 1 μm to about 7 μm, about 1 μm to about 5 μm, or about 1 μm to about 3 μm as measured by NGI. In one embodiment, the aerosolized dry powder composition comprises particles having a FPF of about 40% to about 70%, about 30% to about 60%, or about 50% to about 60% as measured by NGI.

[0168] In some embodiments, PH, PAH, or PPH patients treated by the disclosed methods exhibit one or more of the following treatment responses: (1) a decrease in pulmonary vascular resistance index (PVRI) relative to pre-treatment value, (2) a decrease in mean pulmonary artery pressure relative to pre-treatment value, (3) an increase in hypoxemia score relative to pre-treatment value, (4) a decrease in oxygenation index relative to pre-treatment value, (5) an improvement in right heart function compared to pre-treatment, and (6) an improvement in exercise capacity compared to pre-treatment (e.g., as measured by a six-minute walk test).

[0169] In one embodiment of the disclosed method, the dry powder composition is applied once a day to the PH, PAH or PPH patient. In another embodiment of the disclosed method, the dry powder composition is applied twice a day to the PH, PAH or PPH patient. In still another embodiment of the disclosed method, the dry powder composition is applied three or more times a day to the PH, PAH or PPH patient. In one embodiment, the application is with food. In one embodiment, each application includes 1 to 5 doses from the DPI, such as 1 dose (1 spray), 2 doses (2 sprays), 3 doses (3 sprays), 4 doses (4 sprays) or 5 doses (5 sprays). In one embodiment, the DPI is very small and can be carried by the patient. In one embodiment, the dry powder inhaler is a single-dose dry powder inhaler.

[0170] On the other hand, the present disclosure provides a method for treating pulmonary fibrosis in a patient in need thereof. The method includes administering an effective amount of a dry powder composition disclosed herein, i.e., a dry powder composition comprising a compound of formula (I) or (II) or its enantiomer, diastereomer, or pharmaceutically acceptable salt, to the patient's lungs by inhalation via a dry powder inhaler. In one embodiment, the dry powder composition comprises a compound of formula (I) or (II) or a pharmaceutically acceptable salt thereof. In another embodiment, the dry powder composition comprises a compound of formula (I) or (II). In one embodiment, the administration includes atomizing the dry powder composition with a DPI to form an atomized dry powder composition, and administering the atomized dry powder composition to the patient's lungs via the DPI. In some embodiments, the atomized dry powder composition comprises particles having an MMAD of 1 μm to about 10 μm, about 1 μm to about 7 μm, about 1 μm to about 5 μm, or about 1 μm to about 3 μm as measured by NGI. In one embodiment, the aerosolized dry powder composition comprises particles having an FPF of about 40% to about 70%, about 30% to about 60%, or about 50% to about 60% as measured by NGI. In one embodiment, the dry powder composition is applied to the patient once a day, twice a day, or three times a day or more. In one embodiment, the application is with food. In one embodiment, each application includes 1 to 5 doses from the DPI, such as 1 dose (1 spray), 2 doses (2 sprays), 3 doses (3 sprays), 4 doses (4 sprays) or 5 doses (5 sprays). In one embodiment, the DPI is very small and can be carried by the patient. In one embodiment, the dry powder inhaler is a single-dose dry powder inhaler. Example

[0171] The present invention is further illustrated by reference to the following examples. However, it should be noted that these examples, like the embodiments described above, are illustrative and are not to be construed as limiting the scope of the present invention in any way. Example 1 - Preparation of an inhalable dry powder formulation comprising a compound of formula (II) (treprostinil palmitate) and characterization

[0172] This example describes a mannitol and trehalose based dry powder formulation comprising treprostinil palmitate represented by formula (II), which was prepared by spray drying using a Buchi B-290 spray dryer equipped with an inert gas loop condenser B-295 and a dehumidifier B-296, as well as the characterization and stability testing of the formulation.

[0173] A mannitol-based Treprostinil palmitate dry powder formulation with Treprostinil palmitate / DSPE-PEG2000 / mannitol / leucine components (1 / 0.5 / 80 / 20, 1.5 / 0.75 / 80 / 20, 2 / 1 / 80 / 20, w / w) was successfully prepared. The feed stock solution was prepared by dissolving all components in a 1-propanol / H2O co-solvent system (50 / 50, v / v) without the addition of ammonium bicarbonate. The spray drying yield for the mannitol-based dry powder was greater than 90%. The collected dry powder had spherical particles, a crystalline XRD pattern, and a low moisture content.

[0174] Trehalose-based Treprostinil palmitate dry powder formulations with Treprostinil palmitate / DSPE-PEG2000 / trehalose / leucine components (1 / 0.5 / 80 / 20, 1 / 0.5 / 70 / 30, 1.5 / 0.75 / 80 / 20, 2 / 1 / 80 / 20, w / w) were produced by spray drying a feed stock solution containing all components dissolved in a 1-propanol / H2O co-solvent system (50 / 50, v / v) without the addition of ammonium bicarbonate. The trehalose-based dry powder contained collapsed particles, revealing crystalline leucine and amorphous trehalose. The trehalose-based dry powder showed good physical stability over 3 months. Materials and methods 1. Materials

[0175] Phosphate buffered saline: PBS, pH 7.4, catalog number 10010 (Life technologies) or equivalent

[0176] Sodium chloride: ACS reagent (JT Baker, catalog number 3628-05) or equivalent

[0177] Treprostinil palmitate, formula II, as above

[0178] DSPE-PEG2000: N-(methylpolyoxyethyleneoxycarbonyl)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine, sodium salt, DSPE-020CN (NOF, Tokyo, Japan) or equivalent

[0179] D-Lactose, monohydrate, (Sigma)

[0180] L-Leucine, (Sigma)

[0181] Ammonium bicarbonate (Sigma)

[0182] Anhydrous ethanol (Fisher Sci)

[0183] 1-Propanol (Fisher Sci) 2. Equipment

[0184] Buchi B-290 spray dryer with inert gas loop condenser B-295, dehumidifier B-296, two-fluid nozzle ID 0.7 mm and high performance cyclonic separator (Buchi).

[0185] SEM: Zeiss-Sigma FE-SEM (Germany)

[0186] XRD: (PANalytical, Netherlands)

[0187] DSC 250, TA Instruments, New Castle, DE, USA.

[0188] Tap density tester, JV 1000, (Copley Scientific, UK)

[0189] NGI: Next Generation Impactor, (MSP Corporation, Minnesota, USA)

[0190] PSD:RODOS / M,(Sympatec, Germany)

[0191] Karl Fischer titrator: Aquastar, AQV33, EMD.

[0192] DLS: Atlas, (Wyatt Technology, Santa Barbara, California)

[0193] High performance liquid chromatograph: Waters Alliance 2695 model. HPLC software: Waters Empower TM 3

[0194] Magnetic stirring plate 3. Preparation of a dry powder formulation containing treprostinil palmitate, DSPE-PEG2000, trehalose and leucine in a weight ratio of 1:0.5:80:20 Preparation of stock solution:

[0195] Treprostinil palmitate: 10 mg / mL in 1-propanol

[0196] DSPE-PEG2000: 10 mg / mL in 1-propanol

[0197] Trehalose: 150 mg / mL in deionized water

[0198] Leucine stock solution: 20 mg / mL in deionized water Preparation of feed solution for spray drying:

[0199] The spray drying feed solution was prepared at a weight ratio of 1:100 of Treprostinil palmitate to the sum of trehalose and leucine according to Table 1. The final feed solution had 50% 1-propanol and 20.3 mg / mL solids (Table 2).

[0200] First, trehalose and leucine stock solutions were added to the aqueous phase, followed by the addition of 1-propanol and sonication in a water bath. Then, treprostinil palmitate (Formula (II)) and DSPE-PEG2000 were added, respectively. Stirring was applied throughout the process. contain Treprostinil palmitate / DSPE-PEG2000 / trehalose / leucine (1 / 0.5 / 80 / 20) based on sea Spray drying process of dry powder of algae sugar:

[0201] Spray drying was performed using a Buchi B-290 spray dryer under the following parameters: 150°C inlet temperature, 64°C outlet temperature, 414 L / h spray air flow (36 mm, rotor height), 35% m 3 / h suction rate and 7.5mL / min (22%) feed rate. Table 3 summarizes the process parameters. 4. Preparation of a dry powder formulation comprising treprostinil palmitate, DSPE-PEG2000, mannitol and leucine in a weight ratio of 1:0.5:80:20 Preparation of stock solution:

[0202] Treprostinil palmitate: 10 mg / mL in 1-propanol

[0203] DSPE-PEG2000: 10 mg / mL in 1-propanol

[0204] Mannitol: 150 mg / mL in deionized water

[0205] Leucine stock solution: 20 mg / mL in deionized water Preparation of spray drying feed solution:

[0206] A spray drying feed solution was prepared at a weight ratio of 1:100 of Treprostinil palmitate to the sum of mannitol and leucine according to Table 4. The final feed solution had 50% 1-propanol and 20.3 mg / mL solids (Table 5).

[0207] Mannitol and leucine stock solutions were first added to the aqueous phase, followed by the addition of propanol and sonication in a water bath. Then, treprostinil palmitate and DSPE-PEG 2000 were added separately. Stirring was applied throughout the process. The base consists of treprostinil palmitate / DSPE-PEG2000 / mannitol / leucine (1 / 0.5 / 80 / 20). Spray drying process of mannitol powder:

[0208] Spray drying was performed using a spray dryer Buchi B-290 under the following parameters: 135°C inlet temperature, 60°C outlet temperature, 414 L / h spray air flow (36 mm, rotor height), 35% m 3 / h suction rate and 7.5mL / min (22%) feed rate. Table 6 summarizes the process parameters. 5. Characterization of dry powder Surface Electron Microscopy (SEM)

[0209] The dry powder sample (as received) was poured onto a carbon tape and then coated with 20 nm of gold (Au) using an Electron MicroscopySciences (EMS150T ES) sputter coater. The particle morphology was observed using a field emission scanning electron microscope (FE-SEM) using a Zeiss-Sigma FE-SEM (Germany) at an operating voltage of 5 keV. The working distance was maintained between 8 and 10 mm to obtain a relatively high resolution. X-ray diffraction test (XRD)

[0210] The dry powder samples (as received) were loaded into a zero-background sample holder and then evaluated for structural features using X-ray diffraction (XRD) using a PANalytical (Netherlands) X'Pert diffractometer at 45 kV and 40 mA using a Cu Kα Irradiation was performed at a scan rate of 0.04 rad° / min. The scan range was between 4° and 40° degrees (2θ) with a step time of 97.92 seconds and a step length of 0.0131°. Differential Scanning Calorimetry (DSC)

[0211] About 5-10 mg of dry powder was weighed into the DSC sample pan, which was then sealed airtight. The test was performed as follows: equilibrate at 20°C, adjust the temperature by 0.32°C for 60 seconds, isotherm for 1.0 min, and increase the temperature to 180.0°C at 5°C / min. According to laser diffraction Particle size distribution (PSD)

[0212] Place about 15mg-20mg of dry powder into the desired glass tube. Use Sympatech-HOLOS-REDOS mode. Test as follows: Measuring range R1: 0.1 / 0.18…35μm Trigger conditions Rodos Standard Trigger Diffuser Rodos(RO-AS-15mm) Diffuser Type RODOS / M syringe 4mm with 0 cascade elements Initial pressure 0.5bar Moisture content test (Karl Fischer method)

[0213] The moisture content in the dry powder was analyzed using the Karl Fischer method. Approximately 30 mg of sample was weighed and transferred to a titration vessel. The equipment, materials and operating parameters used were as follows: Materials / Equipment: (1) Titrator: Aquastar AQV33 Karl Fischer titrator equipped with a 5 mL burette and associated balance (2) Balance: An analytical balance capable of weighing up to 4 decimal places was placed with an interface capable of connecting to the Aquastar titrator. (3) Water quality standard 1% NIST (4) Indicator 100% desiccant or 4A molecular sieve 1 / 16 ball (5) Lint-free cloth, Kimwipe (6) Weighing boat (7) Syringe 3mL Solution: (1) Titrant: Aquastar CombiTitrant 2 (2) Solvent: 60 / 40 methanol / formamide Instrument parameters and conditions: (1) Drift: <50μg / min (2) Stirring speed: 40% (3) Mixing time: 300 seconds (4) Termination: Relative drift Bulk density and tap density

[0214] Density test was performed by tap density tester JV 1000 (Coply Scientific, UK). The following procedure was followed. Clean the glass tube and dry it with compressed air; weigh the glass tube and record it as W1; transfer the dry powder to the glass tube, mark the height as A, and record the weight as W2; seal the top with sealing film; put the glass tube into a 5 mL measuring cylinder and tap for 10 min; mark the height after tapping as B; remove the powder from the tube, clean and dry it with compressed air; weigh the glass tube and record it as W3; add water to the scale B and record the weight as W5; add water to the scale A and record the weight as W4. (Assume that the density of water is equal to 1 g / mL) Dry powder loading W2-W1 Bulk density volume (W4-W3) / 1 g / mL Tap density volume (W5-W3) / 1 g / mL Bulk density Dry powder loading / bulk density volume Tap density Dry powder loading / tap density volume Aerodynamic Particle Size Distribution (APSD) measured using NGI

[0215] About 20 mg powder filled in No. 3 Vcaps HPMC capsules was dispersed into the next generation cascade impactor (NGI) (CopleyScientific, UK) running at a volume flow rate of 60 L / min by a commercial inhaler (PlastiapeRS01) and operated for 4 seconds. The drug content collected from each stage of the NGI device was evaluated by HPLC-MS. Fine particle fraction (FPF) was defined as the drug mass (<5 μm) deposited in the NGI divided by the discharged dose. HPLC assay

[0216] Treprostinil (TRE) and Treprostinil palmitate concentrations were determined using a Waters Alliance 2695 model equipped with a PDA detector (Waters 2996) and a Corona charged aerosol detector (Thermo Fisher Scientific). Chromatographic column: ACE 3 C8 HPLC column 4.6x50 (Mac-Mod Analytical, catalog number ACE1120546) Column temperature: 25°C; Mobile phase A: acetonitrile 25%, methanol 25%, water 50%, formic acid 0.1%, triethylamine 0.01% Mobile phase B: acetonitrile 50%, methanol 50%, formic acid 0.1%, triethylamine 0.01% Flow rate: 1 mL / min Measuring the gradient of TRE: Injection volume: 50 μL UV wavelength: 270 ± 2.4 nm Dissolve samples and standards in 33% acetonitrile, 33% methanol, and 33% water. Calibration by fitting the power function Log(area) = A + B*Log(concentration) Retention time: TRE is about 1.8 min, C16TR is about 7.6 min Total recording time: 9 min result 1. Batches of Treprostinil Palmitate Dry Powder Based on Mannitol

[0217] Different batches of Treprostinil palmitate dry powder based on mannitol were prepared by spray drying. In these batches, the amount of Treprostinil palmitate in the dry powder changed from 1% to 5% (weight ratio, w / w), while the ratio of Treprostinil palmitate to DSPE-PEG2000 remained unchanged at 2 to 1. The leucine content was between 0 and 30% (w / w) of the dry powder. The effect of ammonium bicarbonate was also studied. During the spray drying process, the inlet temperature changed from 120°C to 150°C. Table 7A shows the composition and inlet temperature of Treprostinil palmitate dry powder based on mannitol in different batches. For each batch, the amount of Treprostinil palmitate, DSPE-PEG2000, mannitol and leucine was indicated by weight ratio. The amount of Treprostinil palmitate and leucine was also indicated by its approximate weight percentage shown by its ratio in the weight ratio. Table 7B shows the target weight percentages of Treprostinil palmitate, DSPE-PEG2000, mannitol, and leucine in each batch calculated based on weight ratios. 1.1. Effect of leucine content on spray drying recovery

[0218] The effect of leucine on the properties of mannitol-based Treprostinil palmitate dry powder was evaluated. Four leucine loadings were evaluated, 0%, 10%, 20% and 30%. An increase in mannitol loading was applied to compensate for the decrease in leucine content.

[0219] The spray drying recovery (%) of the batch without leucine was low. As the leucine level increased to 10% and 20%, the recovery increased significantly. When the leucine content was further increased to 30%, the recovery decreased slightly ( Figure 1 The batch containing 20% ​​leucine had the highest powder density value (Table 8). 1.2 Effect of leucine content (30%, 20%, 10% and 0%) on powder morphology,

[0220] SEM was performed to examine the effect of leucine content on the powder surface ( Figure 2A-2D). The variation of leucine content resulted in different weight ratios of mannitol to leucine, namely 70 / 30, 80 / 20, 90 / 10 and 100 / 0.

[0221] Treprostinil palmitate dry powder samples with different mannitol to leucine weight ratios (70 / 30, 80 / 20, 90 / 10 and 100 / 0) were produced. SEM data showed that increasing the leucine content from 20% to 30% resulted in a powder with a wrinkled surface ( Figure 2A and Figure 2B ). The dry powder without leucine broke after spray drying and the recovery rate was low ( Figure 2D ). In further studies, 20% leucine was used. 1.3. Effect of leucine content on PSD (tested by laser diffraction)

[0222] Three batches of mannitol-based dry powder containing 10%, 20% and 30% leucine (w / w) were studied by laser diffraction. The batch information is shown in Table 9.

[0223] like Figure 3 As shown, the formulation containing 20% ​​leucine had the smallest particle size (D50). A formulation containing 20% ​​leucine was used in further studies in this example. 1.4. Effects of different amounts of treprostinil palmitate on MMAD

[0224] Different amounts of Treprostinil palmitate were incorporated into mannitol-based dry powder formulations in order to study their effects on dry powder properties. As shown in Table 10, five batches of dry powder were prepared with Treprostinil palmitate ranging from 1% to 5% and a weight ratio of Treprostinil palmitate / DSPE-PEG2000 of 2:1.

[0225] The value of MMAD was constant when treprostinil palmitate was 1% to 2% in the dry powder and increased significantly when treprostinil palmitate was increased to 3% to 5% ( Figure 4 ). 1.5. Effect of spray drying inlet temperature at 150°C, 135°C and 120°C on powder morphology

[0226] At different inlet temperatures, namely 150°C, 135°C and 120°C, mannitol-based Treprostinil palmitate dry powders with the same component ratios were produced (Table 11). The effect of inlet temperature on the morphology of the dry powder was first studied. SEM revealed that there was less surface damage in the dry powder samples spray dried at lower inlet temperatures of 135°C and 120°C ( Figure 5A-5C Since no significant difference was found between 135℃ and 120℃ ( Figure 5B and Figure 5C ), therefore an inlet temperature of 135 °C was used in further studies. 1.6. Effect of ammonium bicarbonate (ABC) in the feed stock solution on the morphology of mannitol-based treprostinil palmitate dry powder

[0227] The effect of ABC on the morphology of mannitol-based Treprostinil palmitate dry powders was examined by adding or not adding ABC to the feed stock solution when preparing the dry powder (Table 12). No changes in the powder surface were observed with the addition of ABC ( Fig. 6A and Figure 6B ). Therefore, ABC was not applied in the mannitol-based Treprostinil palmitate dry powder. 1.7. Physicochemical properties of mannitol-based treprostinil palmitate dry powder

[0228] The mannitol-based Treprostinil palmitate dry powder is produced by spray drying a solution containing all components. It is expected that the mannitol-based dry powder will show some amorphous characteristics, such as Tg in DSC test and broad peaks in powder X-ray diffraction test (XRD). Fig. 7A and Figure 7B DSC and XRD data from batch SD-NNP-179 with 1% treprostinil palmitate and 20% leucine are shown, respectively. No Tg was detected from this batch and a sharp peak was observed in the powder XRD. These two features were also noted in other batches regardless of differences in composition and spray drying conditions. 2. Trehalose-based Treprostinil Palmitate Dry Powder Batch

[0229] Different batches of Treprostinil palmitate dry powder based on trehalose were prepared using spray drying. Table 13A shows the batch composition and inlet temperature for the spray drying process. For each batch, the amount of Treprostinil palmitate, DSPE-PEG2000, trehalose and leucine is indicated by weight ratio. Table 13B shows the target weight percentage of Treprostinil palmitate, DSPE-PEG2000, trehalose and leucine in each batch calculated based on the weight ratio. The Treprostinil palmitate content of the batch varies from 1% to 2% (weight ratio, w / w), while the ratio of Treprostinil palmitate to DSPE-PEG2000 remains unchanged at 2 to 1. The leucine content in the batch is 20% or 30% (w / w). During the spray drying process, the inlet temperature varies from 110°C to 155°C. 2.1. Effect of leucine content on powder morphology

[0230] Trehalose-based Treprostinil palmitate dry powders with two leucine levels (20% and 30%) were prepared (Table 14). SEM data showed that increasing the leucine content from 20% to 30% resulted in wrinkling of the powder surface ( Fig. 8A and Figure 8B ). 2.2. Effect of leucine content on powder aerosol properties

[0231] We incorporated two levels of leucine (i.e., 20% and 30%) into trehalose-based Treprostinil palmitate dry powders and compared their particle size (D50, laser diffraction) and MMAD (Table 14).

[0232] The data in Table 14 show that the dry powder with 30% leucine has a larger geometric particle size (D50) and lower deposition of the powder on the intake line and pre-separator than the dry powder with 20% leucine. The low solubility of leucine will cause leucine to precipitate first. Higher amounts of leucine will trigger precipitation faster, resulting in larger particles. However, there is no significant difference in MMAD. 2.3. Effect of treprostinil palmitate content on dry powder aerosol performance.

[0233] Different amounts of Treprostinil palmitate were incorporated into Trehalose-based Treprostinil palmitate dry powders in order to study their effects on dry powder properties. As shown in Table 15, four batches of dry powders were prepared with Treprostinil palmitate contents ranging from 1% to 2% and the weight ratio of Treprostinil palmitate / DSPE-PEG2000 was fixed at 2:1.

[0234] With more treprostinil palmitate in the dry powder, MMAD and deposition on the intake line and pre-separator increased, while fine particle fraction (FPF) decreased (Table 15). 2.4. Effect of spray drying inlet temperature on powder morphology

[0235] The inlet temperature during spray drying is expected to affect the properties of the dry powder, such as moisture content, particle size and powder morphology. Two inlet temperatures of 130°C and 150°C were investigated using the trehalose based vehicle dry powder batches shown in Table 16.

[0236] SEM showed that the high inlet temperature of 150 °C caused the powder to crack ( Figure 9A-9C ). However, to prevent high moisture in the final dry powder, 150°C will be used for the trehalose based dry powder containing Treprostinil palmitate. 2.5. Physicochemical properties of trehalose-based dry powder

[0237] Similar to the mannitol-based Treprostinil palmitate dry powder, the trehalose-based Treprostinil palmitate dry powder is produced by spray drying a solution containing all components. The dry powder is expected to show some amorphous characteristics, such as Tg in DSC and broad peaks in powder X-ray diffraction (XRD).

[0238] DSC and XRD were performed on the batches of trehalose-based treprostinil palmitate dry powders shown in Table 17. In the DSC test, Tg was observed for all batches, which ranged from 64°C to 80°C. If the powder undergoes a second drying via overnight lyophilization, Tg may increase, as observed in batches SD-NNP-162 and SD-NNP-163, due to the reduction in moisture in the dry powder ( Fig. 10A ). Compared with the mannitol-based dry powder, the XRD of the trehalose-based dry powder showed fewer sharp peaks ( Fig. 10B ), which is attributed to the amorphous state of trehalose. Regardless of the weight ratio of the components and the difference in spray drying conditions, all batches showed similar XRD data. Table 17 shows additional properties of the dry powder batches, including MMAD, FPF and inlet line + pre-separator deposition. 3. Dynamic Vapor Sorption (DVS) Spectra of Dry Powders Based on Mannitol and Trehalose

[0239] Moisture may be introduced into dry powder formulations during spray drying, packaging, and storage, leading to product instability and packaging problems. After spray drying, the moisture in the dry powder can be reduced by secondary drying. However, during packaging, even under humidity-controlled conditions, the powder may absorb moisture when exposed to the environment. The moisture absorption of treprostinil palmitate dry powders based on mannitol and trehalose was examined.

[0240] like Fig.11As shown in Figure 2, when RH% increases from 0 to 40%, the mannitol-based Treprostinil palmitate dry powder can absorb up to 0.3% moisture. Compared with the lactose-based Treprostinil palmitate dry powder, the mannitol-based dry powder absorbs much less moisture, which may be because the mannitol-based dry powder contains crystalline mannitol and leucine, and the stable form of both is the unhydrated form.

[0241] The moisture absorption spectrum of trehalose-based dry powder is shown in Fig.12 (powder with 20% leucine) and Fig.13 (Powder with 30% Leucine). The weight change of the trehalose based dry powders peaks at 50% RH%. After that, the moisture absorption decreases due to the change in the physical form of trehalose from amorphous to crystalline. The powder formulations containing 20% ​​and 30% Leucine have similar moisture absorption data, but the latter formulation has a 1% lower absorption. However, the difference is significant during desorption. The powder containing 30% Leucine exhibits a higher moisture residual at 0% RH%. 4. Stability test of treprostinil palmitate dry powder based on mannitol

[0242] The stability test of the mannitol-based Treprostinil palmitate dry powder was conducted at 40°C without humidity control for 3 months. Five batches of mannitol-based Treprostinil palmitate dry powder containing 1%, 1.5%, 2%, 3% and 5% Treprostinil palmitate were studied (Table 18). The NGI test showed that, in general, the MMAD of the dry powder rose sharply at the 2-month time point and then fell back to a level similar to that at the 1-month time point ( Fig.14 ). SEM data showed that dense small fibers grew on the powder surface ( Fig.15A , Fig. 15B , Fig.16A , Fig. 16B , Fig.17A , Fig. 17B , Fig.18A and Fig.18B ).

[0243] Table 19 shows the details of the stability data of the batches of Treprostinil palmitate dry powder based on mannitol. In all batches of mannitol-based dry powders containing 20% ​​leucine, significant changes in MMAD and FPF were observed. In addition, the initial MMAD of the dry powders with 1%, 1.5% and 2% Treprostinil palmitate was lower than that with 3% and 5% Treprostinil palmitate. 5. Stability test of trehalose-based dry powder

[0244] Stability studies were also conducted for trehalose-based dry powder formulations with 20% or 30% leucine and treprostinil palmitate in the range of 1% to 2% under the same conditions as for the mannitol-based dry powder formulations. No significant changes were observed over the 3.5 month study period (Tables 20 and Fig.21A , Fig. 21B , Fig.22A , Fig. 22B , Fig.23A , Fig. 23B , Fig.24A and Fig. 24B ).

[0245] Table 21 shows the details of the stability data of the trehalose-based treprostinil palmitate dry powder batches. The MMAD for batches SD-NNP-162 and SD-NNP-188 increased significantly (Tables 21 and Fig.19 The FPF value of batch SD-NNP-162 decreased most significantly ( Fig. 20 ). Batch SD-NNP-163 (Treprostinil palmitate 1%, Leucine 30%) showed the lowest and stable MMAD and the highest stable FPF. Summary of Inspection Results for Mannitol-Based Treprostinil Palmitate Dry Powder Formulations

[0246] In mannitol-based Treprostinil palmitate dry powder, the addition of leucine resulted in high spray drying recovery. The mannitol-based dry powder with 20% leucine showed spherical particle shape and low geometric diameter (D50 = 2.75 μm).

[0247] During the spray drying process, the inlet temperature varied from 120°C to 150°C did not affect the morphology of the mannitol-based Treprostinil palmitate dry powder. Since the moisture content of the dry powder produced at 135°C was about 1%, the inlet temperature was set at 135°C.

[0248] No glass transition was detected from the mannitol-based Treprostinil palmitate dry powder in the DSC test, which supported the X-ray diffraction test (XRD) results, indicating that it was a crystalline material. In addition, the spray-dried mannitol-based Treprostinil palmitate dry powder had low hygroscopicity, absorbing less than 1% moisture at 90% RH%.

[0249] In the stability testing of mannitol-based Treprostinil palmitate dry powders, the sharp increase in MMAD was at the 2-month time point for formulations with 1% to 2% Treprostinil palmitate, and at the 1-month time point for formulations with higher Treprostinil palmitate. MMAD decreased at later time points. All formulations showed fibers on the powder surface after storage. Summary of the Examination Results of Trehalose-Based Dry Powder Formulations of Treprostinil Palmitate

[0250] Compared with the trehalose-based treprostinil palmitate dry powder containing 20% ​​leucine, the trehalose-based treprostinil palmitate dry powder containing 30% leucine had a wrinkled surface and deposited less powder in the air intake line and pre-separator. However, there was no significant difference in MMAD between the two.

[0251] The inlet temperature during spray drying was selected to be 150 °C to achieve a lower moisture content in the final dry powder.

[0252] Glass transition temperature (Tg) was observed to be in the range of 64°C to 80°C, indicating the amorphous state of trehalose in the dry powder. Trehalose-based treprostinil palmitate dry powder showed higher water absorption compared to mannitol-based treprostinil palmitate dry powder.

[0253] In stability testing, most of the tested trehalose-based Treprostinil palmitate dry powder formulations exhibited no significant change in FPF. All formulations showed hair-like crystals on the powder surface after storage.

[0254] In summary, the data of this example show that the treprostinil palmitate content in the range of up to 2wt% does not affect the physical properties of the treprostinil palmitate dry powder. At 3wt% and 5wt% treprostinil palmitate, an increase in the initial MMAD of the mannitol-based powder was noted. The leucine content was found to be important for the dry powder aerosol properties. Example 2 - Preparation of an inhalable mannitol and trehalose based dry powder formulation of treprostinil palmitate Fabrication, Packaging and Characterization

[0255] This example describes the manufacture of four Treprostinil palmitate dry powder formulations, Formulations A, B, C, and D, by spray drying and encapsulation. Formulations A and D are mannitol-based, and their compositions, expressed in weight ratios and target weight percentages calculated based on the weight ratios, are shown in Table 22. Formulations B and C are trehalose-based, and their compositions, expressed in weight ratios and target weight percentages calculated based on the weight ratios, are shown in Table 23. This example also describes the characterization of the particle size, morphology, moisture content, solvent content, physical state, gas phase adsorption profiles, thermal properties, and weight loss as a function of temperature of Formulations AD. 1. Spray Drying of Formulations A, B, C and D

[0256] Treprostinil palmitate dry powder formulations A, B, C and D were manufactured using a BLD-200 spray dryer, with the incoming solids for each formulation being approximately 55 grams. Between each condition, a blank solvent solution was sprayed to ensure that the previous formulation was cleared from the solution line. The spray dryer was not additionally cleaned between conditions.

[0257] Each of the four formulations was prepared as a separate solution. The solutions were prepared at room temperature and did not need to be protected from light. For each solution preparation, the following steps were performed: 1. Dissolve leucine in deionized water. 2. Dissolve sugar (mannitol or trehalose) in deionized water. 3. The aqueous solution was filtered through a 0.2 μm PVDF membrane. 4. Dissolve DSPE-PEG2000 in 1-propanol. 5. Dissolve Treprostinil palmitate in 1-propanol. 6. Add the organic solution to the stirring aqueous solution.

[0258] The spray drying formulations and process conditions are listed in Table 24. The manufacturing yields ranged from 54% to 80% by mass. The packaging of bulk dry powders of each formulation was carried out in a dry glove box. 2. Powder packaging

[0259] The dry powder formulation is packaged by using Xceldose 600S so that each formulation fills 50-51 capsules. The relative humidity of the room is less than 30%. The summary of the packaging is shown in Table 25. For example, capsules are made from a typical batch of formulation D (containing 1.50wt% treprostinil palmitate, 0.75wt% DSPE-PEG2000, 68.45wt% mannitol and 29.30wt% leucine) in the following manner: 112.5μg treprostinil palmitate, 56.2μg DSPE-PEG2000, 5133.8μg mannitol and 2197.5μg leucine are filled in each capsule. It is observed that the wt% of each component of other batches of formulation D varies independently with the typical wt% value ±5% as shown above or varies independently within the typical wt% value ±5% as shown above to have equivalent characteristics and performance. The capsules were collected in a glass jar and heat sealed in a foil bag with 0.5 g of molecular sieve desiccant. 3. Analytical Characterization

[0260] Each of the four formulations was evaluated for particle size distribution, particle morphology, water content, residual solvents, physical state, moisture sorption, and thermal properties. A summary of the results is shown in Table 26. Particle size distribution

[0261] Since these formulations are targeted for respiratory delivery, the target particle size should be less than 5 μm. Particle size distribution was measured by laser diffraction on a Malvern Mastersizer 2000 with a Scirocco 2000 dry powder dispersion unit. For method development, an initial pressure titration screening (n=1) was performed on all samples and it was observed that the results were almost identical between the three dispersion gas pressures used (2.5, 3.0 and 3.5 bar) ( Fig.25 Based on this initial screening, two additional replicates (n=2) were measured at a dispersion pressure of 3.0 bar. The results were averaged and the distribution is shown in Fig.26 and Table 27. For these measurements, the Fraunhofer approximation model was used. A small sample pan was used with a feed rate of 65%, a background measurement time of 10 seconds, and a sample measurement time of 30 seconds. Masking filtering was enabled to capture data between 1% and 6%. 3.2. Particle morphology

[0262] Each of the four formulations was imaged using a Hitachi SU3500 scanning electron microscope at 500, 1500, and 5000 magnifications. Images taken at 5000 magnification are shown in Fig. 27 (Preparation A), Fig.28 (Preparation B), Fig.29 (Preparation C) and Fig.30 (Formulation D). All formulations contained intact and collapsed spherical particles with a diameter of about 3 μm or less, which is consistent with the laser diffraction results. Formulation C appeared the most wrinkled, while Formulation A appeared the most "smooth". Surface roughness can improve aerosol performance. No obvious crystalline surface formation or particle fusion was observed for all formulations. 3.3. Water content

[0263] Since the preparation is spray-dried from a solvent mixture containing water, the water content of the sample is analyzed using Metrohm874Oven Sample Processor. Before the test sample, three blanks and three water standards are first tested to determine the system suitability. For each preparation, twenty milligrams of sample (n=3 replicates) are heated to 140°C from a starting temperature of 50°C with a heating rate of 2.5°C / min. As shown in Table 28, by mass, the water content of all preparations is less than 3%. The preparations (B and C) containing trehalose show higher water content than those (A and D) prepared with mannitol. The preparations (C and D) containing higher leucine content also show higher water content. 3.4. Residual solvents

[0264] Since the other component in the spray solvent is 1-propanol, the residual amount of 1-propanol was determined using the headspace method on an Agilent 7890 gas chromatograph system. All samples had less than 0.5% 1-propanol by mass (Table 29), and the formulations containing more leucine (C and D) had lower residual 1-propanol, as did the formulations containing mannitol (A and D). Powder X-ray diffraction

[0265] The crystallinity of the samples was evaluated using a Rigaku MiniFlex 600 powder X-ray diffractometer. The samples were prepared on a 0.2 mm zero background holder (ZBH) disk and run on the instrument from 3 to 40 2θ. Formulations A and D exhibited crystalline mannitol (possibly a mixture of polymorphs) and leucine, while formulations B and C exhibited crystalline leucine and amorphous trehalose.

[0266] The components as pure materials can exhibit different diffraction intensities compared to the spray dried formulations. The formulations containing mannitol appear to have similar diffraction patterns. The formulations containing trehalose also appear to have similar amorphous diffraction patterns. 3.6. Differential Scanning Calorimetry (DSC)

[0267] Thermal transitions can be used to predict the stability of formulations. Four formulations were scanned on a TA Instruments Q2000 differential scanning calorimeter. The samples were equilibrated overnight in a dry environment (<5% RH), after which they were airtightly sealed and run on the instrument. The adjustment was set to ±1.5°C / min, and the heating rate from 0 to 180°C was 2.5°C / min. Thermal events were observed at 64°C for all samples, and the event may be related to the melting of Treprostinil palmitate or DSPE-PEG2000.

[0268] No crystallization events were observed for the samples, a positive indication of thermal stability. No glass transition was detected for the mannitol-based formulations (A and D), which supported the powder X-ray diffraction (PXRD) results for the crystalline material. Formulations A and D melted at 164°C, which is consistent with the melting temperature of mannitol. Formulations B and C exhibited a glass transition at 83°C, which may be due to amorphous trehalose. Formulation B also had thermal events at 133°C and 158°C that were not observed for the other samples. Thermogravimetric analysis (TGA)

[0269] The thermal decomposition data of the formulation was measured using a TA Instruments Discovery thermogravimetric analyzer. The sample was run from 0°C to 300°C at a rate of 2.5°C / min. The formulation began to decompose rapidly after 180°C. A weight change was observed at about 100°C, which corresponds to the water content. Dynamic gas phase adsorption

[0270] Adsorption and desorption spectra were measured on a Surface Measurement Systems DVS Advantage 1. The samples were run from 0 to 90% RH at 25°C with a step change of 10% RH. All four formulations showed a weight change event starting at about 50% or 60% RH, depending on the formulation. The samples were run a second time to evaluate the water uptake after crystallization. No changes were observed for any of the samples in the second cycle, and most of the samples did not retain water in the final desorption step.

[0271] The sorption results for Formulation D show a change at higher humidity (about 70% RH) compared to the results for Formulation A. A slight loss of mass was observed for Formulation D at about 50% RH, whereas Formulation A appeared to continue to lose mass even as it sorbed additional moisture from higher humidity.

[0272] The absorption rate of the trehalose formulations showed that at 40% relative humidity, it took approximately 50 to 90 minutes (depending on the level of leucine content) to reach water equilibrium. Lower leucine contents absorbed water faster than higher leucine contents. 3.9. Evaluation of the aerosol properties of formulations A, C and D by aerodynamic particle size distribution (APSD) according to NGI

[0273] Capsules of Treprostinil palmitate dry powder formulations A, C and D were stored at 25°C or 40°C for 1-3 months. There was no change in the appearance of the encapsulated dry powder, such as no browning or obvious growth or hardness changes. The particle size distribution (PSD) of the dry powder formulation measured by laser diffraction was not affected after storage at 25°C or 40°C for 3 months.

[0274] Treprostinil palmitate dry powder formulations A, C, D were packaged and stored at 40°C for 1 month, 2 months or 3 months, or at 25°C for 3 months. The fine particle dose (FPD) of the formulations from the stored capsules and the initial (T=0) FPD of the formulations were measured by NGI. Treprostinil palmitate dry powder formulations A, C and D were also stored in bulk at 40°C or 25°C for 3 months, then filled into capsules and administered on the same day for FPD determination. The FPD results of formulations A, C and D are shown in Fig.31 , Fig.32 and Fig.33 . These data indicate that Formulation D had the least change in FPD (-3.7%) after storage in capsules at 25°C for 3 months. Additionally, Formulation D had a -5.2% change in FPD when stored in bulk at 25°C for 3 months and filled into capsules and administered on the same day. Furthermore, for each of Formulations A, C, and D, storage at 40°C revealed that aerosol performance as measured by FPD could not be predicted upon long-term storage at 25°C. Based on these data, there is no need to condition or pretreat the powder or capsules to alter aerosol performance.

[0275] The expelled dose and total recovery of Formulations A, C and D in the above capsules were also determined, and the results are shown in Table 30.

[0276] The above data show that Formulation D shows minimal changes in expelled dose after storage in capsules at 25°C for 3 months and after storage in bulk at 25°C for 3 months and then filled into capsules and administered on the same day. In summary, based on the changes in aerosol properties, Formulation D appears to be stable at 25°C for at least 3 months. In addition, the latest stability data supports a shelf life of 6 months when stored at 2°C-8°C. Example 3 - Determination of the stability of a trehalose-based dry powder formulation of Treprostinil palmitate under accelerated storage conditions Aerosol properties and pulmonary pharmacokinetic profiles in rats following inhalation of dry powder formulations

[0277] Trehalose-based Treprostinil palmitate dry powder consisting of Treprostinil palmitate, DSPE-PEG2000, Trehalose (Treh) and Leucine (Leu) in a weight ratio of 1:0.5:70:30 was produced by spray drying using a Buchi B-290 system as described in Example 1. The dry powder was stored in sealed glass vials at 40°C and uncontrolled ambient humidity for accelerated stability studies. The aerosol properties of the powder were measured after 1.5, 2.5 and 3.5 months of storage. Methods and Materials 1. Dynamic Vapor Sorption (DVS) Study

[0278] The moisture absorption curve was obtained using a dynamic vapor sorption (DVS) automated gravimetric sorption system (DVS Intrinsic1Plus, Surface Measurement System, Pennsylvania, USA). Approximately 20 mg of powder was loaded and an adsorption / desorption isotherm cycle was performed at 25°C (from 0% relative humidity (RH) to 90% RH to 0% RH, in increments of 10% RH). The change in powder mass with RH (%) was measured and plotted. 2. DPI device and aerodynamic particle size distribution (APSD) test

[0279] RS01 Mod.7 DPI device (High Resistance, code 239700002AA, Plastiape, Italy) was used in this study. The mass median aerodynamic diameter (MMAD) of trehalose-based treprostinil palmitate dry powder was measured using a Next Generation Impactor (NGI) at 60 L / min.

[0280] About 15 mg of Treprostinil palmitate dry powder was loaded into a size 3 HPMC capsule (Qualicaps, Inc.). The capsule was loaded into the DPI device and actuated to characterize the aerosol particle size distribution (APSD). The amount of drug deposited on each impactor stage and filter was analyzed by high performance liquid chromatography (HPLC). 3. Nose-Only Inhalation

[0281] Treprostinil palmitate dry powder was delivered using a dry powder disperser (Vilnius aerosol generator (VAG), CH Technologies, USA) through a 12-port nasal inhalation chamber. About 1 g of dry powder was loaded into the VAG. The VAG was continued at a flow rate of 8 L / min for a total of 20 minutes at 1.0 V. The dry powder delivery system is described in more detail in Li et al., "Inhaled INS1009 Demonstrates Localized Pulmonary Vasodilation," European Respiratory Society (ERS) International Congress, September 3-7, 2016, London, United Kingdom, Abstract No: 853952 (poster PA2845), the contents of which are incorporated herein by reference in their entirety. 4.PK Sample Collection and Analysis

[0282] Rat lung tissue was harvested immediately after delivery (approximately 0.5 h), 6, 12 and 24 h after administration. Lung tissue samples were analyzed for Treprostinil palmitate and Treprostinil (TRE) by LC-MS / MS. Results are expressed as "Treprostinil palmitate equivalents" to take into account the hydrolysis of Treprostinil palmitate postmortem. Treprostinil palmitate equivalent, ng / g = [treprostinil palmitate, ng / g] + [TRE, ng / g]* (MM Treprostinil Palmitate / MM TRE) MM: molar mass result 1. Dynamic Vapor Sorption (DVS)

[0283] The dynamic vapor sorption (DVS) of the Treprostinil palmitate dry powder formulation is shown in Fig.34 When the dry powder is exposed to increasing RH to 50%, the absorbed moisture increases from 0% to more than 8% and is not completely reversible with desorption remaining at 5.5% or higher. To keep the moisture content at or below about 4%, the RH exposure of the powder can be controlled to <30% during the manufacturing process. 2. Aerodynamic Particle Size Distribution (APSD)

[0284] The APSD data of Treprostinil palmitate stored for 1.5 months (T=1.5M), 2.5 months (T=2.5M) and 3.5 months (T=3.5M) at 60 L / min as well as the initial APSD (T=0) are shown in Fig.35 and Table 31. The distributions from all four time points were comparable.

[0285] The fine particle fraction (FPF), MMAD, and geometric standard deviation (GSD) values ​​of the aerosolized dry powder after storage for up to 3.5 months at 40°C and uncontrolled ambient humidity are summarized in Table 32. The FPF values ​​ranged from 54.6% to 58.7%. The MMAD values ​​ranged from 1.25 μm to 1.44 μm. The GSD values ​​ranged from 3.5 to 4.1. 3. Rat lung PK results

[0286] The concentrations of treprostinil palmitate equivalent (treprostinil palmitate plus treprostinil) in the lung after inhalation of aerosolized INS1009 or aerosolized treprostinil palmitate dry powder are summarized in Tables 33 and Fig.36 Aerosolized INS1009 contains treprostinil palmitate at a molar ratio of 45:45:10 suspended in PBS and excipients squalane and DSPE-PEG2000 (see Corboz et al., "Preclinical Pharmacology and Pharmacokinetics of Inhaled Hexadecyl-Treprostinil (C16TR), a Pulmonary Vasodilator Prodrug," J Pharmacol Exp Ther. 363:348-357 (2017), the contents of which are incorporated herein by reference in their entirety). Aerosolized INS1009 and C16TR (treprostinil palmitate) dry powder have similar lung PK profiles after inhalation in rats. The statistical comparison of the two profiles is summarized in Table 34, which shows comparable slopes for the two profiles.

[0287] In conclusion, this study demonstrated that the aerosol particle size distribution of a trehalose-based treprostinil palmitate dry powder formulation was reproducible when stored in sealed vials at 40 °C and uncontrolled RH for up to 3.5 months and that the lung PK profile of the treprostinil palmitate dry powder formulation was comparable to that of aerosolized INS1009. Example 4 - Pharmacodynamics of Treprostinil Palmitate Dry Powder Formulation Based on Mannitol and Trehalose in Rats Kinetic evaluation

[0288] In this study, the lung and plasma pharmacokinetics (PK) of two different Treprostinil palmitate dry powder formulations with mannitol (i.e., Formulation D as described in Example 2) or trehalose (i.e., Formulation C as described in Example 2) as their primary excipients were evaluated. The compositions of Formulations D and C, expressed as weight ratios, target weight percentages calculated based on the weight ratios, and actual weight percentages of components from a typical batch of each formulation are summarized in Tables 35A and 35B, respectively. method

[0289] Male Sprague Dawley rats (300-400 g) were exposed to aerosols of dry powder formulations D or formulation C obtained using a 12-port nose inhalation chamber and a Vilinius aerosol generator (VAG). Rats were placed in restraint tubes attached to nose ports in the chamber. In two independent studies using formulations D and formulation C, 9 rats were used in each study, and 1 port was used to collect aerosol drugs on the filter. A simplified study was also performed using formulation D, in which 6 rats were used, and 1 port was used to collect the amount of drug deposited on the filter.

[0290] For drug exposure, one gram of material is placed in VAG. The output of VAG is set to 1 volt (V), and the drug is dispersed and delivered to the nose chamber with a bias flow of 8L / min. Air enters the bottom of the nose chamber and leaves from the top. The exposure duration is set to 20min. A vacuum source (0.5L / min) is attached to the filter, and the drug sampling time is set to 5min. The amount of Treprostinil palmitate deposited on the filter is measured by HPLC and a charged aerosol detector (CAD). The drug dose delivered is calculated by the drug concentration (from the filter data) inhaled, the exposure duration, the per minute breathing volume and body weight, and the deposition fraction 1.0 for the drug delivered in the nose and the deposition fraction 0.1 for the drug delivered to the lung. Dosage is represented by per kg body weight.

[0291] In two preliminary studies using formulations D and formulation C, blood samples were collected at 0.5, 2, 4, 6, 12 and 24 hours after drug exposure. The blood samples were centrifuged to extract plasma. In these studies, respiratory tissues of the larynx, trachea, carina + bronchi and lungs were collected at 0.5, 6, 12 and 24 hours after drug exposure. In a simplified study using formulation D, blood samples were obtained at 0.5, 2, 4, 12 and 24 hours after drug exposure, and respiratory tissues were harvested at 0.5 and 24 hours after drug exposure. The concentrations of Treprostinil (TRE) in plasma and Treprostinil palmitate and TRE in respiratory tissues were measured by LC-MS / MS. For all respiratory tissues, the concentration of Treprostinil palmitate (C16TR) equivalent (C16TReq) was derived from the Treprostinil palmitate and TRE concentrations (C16TReq = Treprostinil palmitate + [TRE x 615 / 390.5 ng / g]), where 615 and 390.5 are the molecular weights of Treprostinil palmitate and TRE, respectively. The following PK parameters were derived using the PK Solver program in Microsoft Excel using lung Treprostinil palmitate equivalent and plasma TRE data: λz (terminal elimination rate constant), T 1 / 2 、T max , C max and AUC 0-inf . result

[0292] Exposure of rats to Formulation D and Formulation C resulted in a slightly higher total delivered dose of 78 μg / kg body weight in the case of Formulation D compared to 58 μg / kg body weight in the case of Formulation C. Treprostinil palmitate equivalent concentrations in the lungs measured at 0.5 hours post-exposure were also slightly higher with a mean of 3072 ng / g in the case of Formulation D compared to 1711 ng / g in the case of Formulation C. 1. Lung and upper airway treprostinil palmitate, TRE and treprostinil palmitate equivalents

[0293] The highest concentrations of Treprostinil palmitate, TRE and Treprostinil palmitate equivalents in the lung occurred at 0.5 hours post-exposure in both Formulation D and Formulation C. There was a slow monoexponential decline of Treprostinil palmitate and TRE over 24 hours, with concentrations of both Treprostinil palmitate and TRE consistently higher at all time points in the case of Formulation D. These results for Treprostinil palmitate equivalents (C16TReq) are shown in Fig.37 , which shows a slow decrease in lung Treprostinil palmitate equivalent concentrations over 24 hours, with Treprostinil palmitate equivalent concentrations being consistently higher in the case of Formulation D compared to Formulation C. Fig.38 Treprostinil palmitate (C16TR) concentrations in the lung following inhalation of Formulation D or Formulation C are shown. Fig.39 TRE concentrations in the lung after inhalation of Formulation D or Formulation C are shown.

[0294] Comparison of the derived PK parameters of treprostinil palmitate equivalents revealed that λz, T 1 / 2 and T max There was no major difference between Formulations D and C, but in the case of Formulation D there was 79% higher treprostinil palmitate equivalents C max and 130% higher AUC 0-24h (Table 36).

[0295] Regarding the deposition of Treprostinil palmitate in the larynx, trachea, carina + bronchi, and lungs, the majority of Treprostinil palmitate (>97%) was deposited in the lungs. This was observed in both Formulation D and Formulation C. No nasal tissue was collected for this study. 2. Plasma TRE

[0296] The highest concentration of TRE in plasma was observed at 0.5 hours, which declined slowly over 24 hours in the case of Formulation D and declined more rapidly in the case of Formulation C. In the case of Formulation C, plasma TRE concentrations were below detection levels by 24 hours ( Fig.40 ).

[0297] Comparison of the derived PK parameters of plasma TREC revealed a 34% lower λz and 51% higher T1 / 2 in the case of Formulation D compared to Formulation C (Table 37). max 10% higher and AUC 0-t 51% higher (Table 37).

[0298] In conclusion, inhaled treprostinil palmitate dry powder formulations D and C resulted in lower treprostinil plasma C compared with inhaled treprostinil. maxValues ​​and sustained levels of treprostinil in plasma and lung. Comparison of the PK profiles in rats between Formulation D and Formulation C delivered under similar conditions (i.e., 1 V VAG output for 20 min) resulted in a 34% higher delivered dose, i.e., 78 μg / kg, in the case of Formulation D, compared to 58 μg / kg in the case of Formulation C. The concentrations of treprostinil palmitate, TRE, and treprostinil palmitate equivalents in the lungs were consistently higher in the case of Formulation D, which may be due to the higher delivered dose in the case of Formulation D, but both formulations showed a slow monoexponential decline over 24 hours. A similar trend was observed for the higher plasma TRE concentrations in the case of Formulation D, and there was a slow decline over 24 hours in the case of both formulations. Most of the TRE and treprostinil palmitate exposure occurred in the lungs, with less than 3% deposited in the upper respiratory tract regions of the larynx, trachea, and carina + bronchi.

[0299] These results for Formulation D and Formulation C indicate a PK profile similar to that previously observed for aerosolized INS1009, showing peak concentrations of Treprostinil palmitate equivalents in the lung and TRE in plasma by 30 min, with a slow monoexponential decline over 24 hours. However, a difference in lung Treprostinil palmitate equivalents: plasma TRE C was observed between the Treprostinil palmitate dry powder formulations and aerosolized INS1009. max For aerosolized INS1009, typical pulmonary treprostinil palmitate equivalents: plasma TRE C max The ratio is about 800, while for the Treprostinil palmitate dry powder formulation, the ratio is in the range of about 1,600-13,000. See Corboz et al., J Pharmacol Exp Ther 363: 1-10 (2017), the contents of which are incorporated herein by reference in their entirety. Since pulmonary vasodilation is more related to the local activity of TRE in the lungs and less related to TRE levels in plasma (see Chapman RW et al., Pulm. Pharmacol. Ther. 49: 104-111 (2018), the contents of which are incorporated herein by reference in their entirety), the ratio of pulmonary Treprostinil palmitate equivalent: plasma TRE C max The unexpected difference in the ratios suggests that administration of the Treprostinil palmitate dry powder formulation beneficially results in lower systemic exposure to TRE, thereby minimizing potential systemic adverse events, such as a decrease in systemic blood pressure. Example 5 - Effect of mannitol and trehalose-based treprostinil palmitate dry powder formulation on hypoxic challenge Evaluation of the efficacy of telemetry in rats

[0300] This example describes the in vivo efficacy evaluation of two different Treprostinil palmitate dry powder formulations: Mannitol-based Formulation D and Trehalose-based Formulation C (Table 35A and Table 35B) described in Examples 2 and 4. Efficacy was determined in rats prepared by implanting a telemetric probe into the right ventricle to measure the inhibition of the increase in right ventricular pulse pressure (RVPP) induced by exposure to an inhaled hypoxic gas mixture. method

[0301] Experiments were conducted in male Sprague Dawley rats implanted with telemetry probes in the right ventricle and descending aorta to measure RVPP and mean systemic arterial blood pressure (mSAP). These cardiovascular parameters were measured while breathing normoxic air (21% O2 / N2), then exposed to hypoxic air (10% O2 / N2) for 10 min and returned to breathing normoxic air. In each experiment, the RVPP increase (ΔRVPP caused by hypoxia) caused by hypoxia challenge was measured before drug exposure and 1, 6, 12 and 24 hours after exposure to inhaled Treprostinil palmitate dry powder formulation D or formulation C. The drug formulation was administered with a Vilinius aerosol generator (VAG) at 1 volt (V) output and delivered to a 12-port nasal inhalation chamber for 20 min. The aerosol was dispersed from the VAG and delivered to the bottom of the nasal inhalation chamber at a bias flow of 8 L / min. The filter was connected to one of the nose ports and attached to a vacuum flow of 0.5 L / min for 5 min. The amount of Treprostinil palmitate deposited on the filter was measured by LC-MS / MS. The concentrations of Treprostinil palmitate and / or TRE were analyzed in plasma and respiratory tissue samples of rats. result

[0302] Exposure of rats to inhaled hypoxia increases RVPP by 10-13 mmHg above normoxic values. At the end of the hypoxic challenge, RVPP decreases immediately within a few minutes and returns to pre-hypoxic values ​​within 10 minutes. Drug effects are determined by comparing the delta RVPP due to hypoxia at multiple time points up to 24 hours after drug exposure to the combined values ​​of 2-3 measurements obtained before drug exposure.

[0303] The experimental results are shown in Fig.41 .exist Fig.41In the table, values ​​are mean ± SEM (n = 6-8 rats for Formulation D; and 4 rats for Formulation C). Day -1 represents the baseline value before drug exposure, and Day 0 represents the value after drug exposure. △RVPP represents an increase in right ventricular pulse pressure. "*" indicates P < 0.05 compared to the combined value before drug exposure on Day -1 (1, 6 and 12 hours). The results showed that exposure to Formulation D reduced the △RVPP caused by hypoxia, with statistically significant (P < 0.05) inhibition observed at 1, 6, 12 and 24 hours. Similar results were obtained with Formulation C, with statistically significant (P < 0.05) inhibition observed at 1, 6, 12 and 24 hours. By 24 hours after treatment with Formulation D and Formulation C, RVPP tended to return to the baseline value observed before drug exposure. As a control, exposure to an inhaled mannitol vehicle (containing a target weight percentage of 69.48 wt% mannitol, a target weight percentage of 29.78 wt% leucine, and a target weight percentage of 0.74 wt% DSPE-PEG2000) did not inhibit the ΔRVPP response to hypoxia at 1, 6, 12, and 24 hours after administration (data not shown). Based on the concentration of the inhaled drug, the duration of exposure, body weight, respiratory volume per minute, and a deposition fraction of 1.0, the delivered dose of Formulation D was 78 μg / kg body weight (treprostinil palmitate equivalent concentration of 3072 ng / g), and the delivered dose of Formulation C was 58 μg / kg body weight (treprostinil palmitate equivalent concentration of 1711 ng / g). See Alexander DJ et al., Inhal. Toxicol. 20: 1179-1189 (2008), the contents of which are incorporated herein by reference in their entirety.

[0304] In summary, the efficacy evaluation in telemetry rats challenged with hypoxia showed that inhaled treprostinil palmitate dry powder formulations C and D inhibited RVPP increases for up to 24 hours after drug exposure. By 24 hours, the RVPP response to hypoxia tended to return to the baseline value observed before exposure to the drug. In contrast, aerosolized INS1009 inhibited hypoxia-induced RVPP increases for up to 12 hours at a delivered dose of 76 μg / kg, and returned to baseline values ​​by 24 hours. Inhaled treprostinil inhibited hypoxia-induced RVPP increases within 2 hours at the highest dose (215 μg / kg) used, and the inhibition lasted for 1 hour at lower inhaled doses (15, 53 or 110 μg / kg). These results demonstrate the prolonged inhibition of hypoxia-induced RVPP increases by treprostinil palmitate dry powder formulations in telemetry rats. Example 6 - Effect of a dry powder formulation of Treprostinil palmitate based on mannitol and trehalose on cough in guinea pigs Assessment of the effects of cough and ventilation

[0305] This example describes a study of the effects of mannitol-based Treprostinil palmitate dry powder Formulation D and trehalose-based Treprostinil palmitate dry powder Formulation C (described in Examples 2 and 4; see formulation compositions in Tables 35A and 35B) on the production of cough, changes in ventilation, and changes in Penh (a dimensionless index of altered breathing patterns typically seen during bronchoconstriction in conscious male guinea pigs). method

[0306] Experiments were performed in male Hartley guinea pigs. After a 3-day adaptation period, the guinea pigs were placed in a whole-body plethysmograph to measure ventilation (tidal volume, respiratory rate, and minute ventilation), Penh, and cough using established techniques. See Corboz et al., J Pharmacol Exp Ther 363: 1-10 (2017); Chong BTY et al., J.Pharmacol.Toxicol.Methods 39, 163-168 (1998); Lomask, Exp.andToxicol.Pathol.57, 13-20 (2006), each of which is incorporated herein by reference in its entirety. Cough was measured by plethysmograph recordings showing a large amount of inspiration followed by a large amount of exhalation, and confirmed by manual observation, video recording, and cough sounds. Ventilation, Penh, and cough data were measured during a 15 min baseline period before exposure to dry powder aerosols. The test article is then delivered as a dry powder for 15min, followed by a 120min observation period after the aerosol compound is applied, wherein the test article includes preparation D, preparation C and their respective vehicles, i.e., mannitol vehicle (containing a target weight percentage of 69.48wt% mannitol, a target weight percentage of 29.78wt% leucine and a target weight percentage of 0.74wt% DSPE-PEG2000) and trehalose vehicle (containing a target weight percentage of 69.48wt% trehalose, a target weight percentage of 29.78wt% leucine and a target weight percentage of 0.74wt% DSPE-PEG2000). The air delivered for the aerosol of all steps of the experiment is supplied by an air compressor. As measured, the typical humidity of the air supplied is about 30%. Ventilation, Penh and cough are measured before, during and after exposure to the test article. Atomization test articles are produced using a Vilnius aerosol generator (VAG). The air flow rate through the VAG is set to 4.5L / min to disperse the aerosol, and merged with 1L / min of humidified air (30% humidification) to promote the aerosol to be delivered to the plethysmograph and minimize the problem of static adhesion. Therefore, the total inflow of air is 5.5L / min. The generator output from the VAG is 1, 0.75 and 0.5 volts, wherein each VAG output continues 15min to deliver 3 different drug doses, and delivers a higher dose at a higher voltage. A vacuum flow of 8L / min is established at the bottom of the plethysmograph so that air and aerosol enter at the top of the system and leave at the bottom of the system. A separate 0.5L / min vacuum source is connected to a filter, and the filter is attached to the port of the plethysmograph to sample the drug (treprostinil palmitate) concentration.Filter sampling was maintained throughout the study; i.e., 135 min (excluding the 15 min exposure time) was used to calculate the aerosol concentration of the drug in the nasal chamber and the total delivered drug dose inhaled. Filter samples were analyzed for Treprostinil palmitate concentrations. At the end of the study, guinea pigs were euthanized and blood (plasma), lungs, trachea, larynx, and carina + bronchi were collected to measure Treprostinil palmitate and TRE concentrations in these samples. result

[0307] Exposure to Formulation D, Formulation C, or the mannitol and trehalose vehicle was well tolerated and did not result in any mortality.

[0308] A nebulized formulation D (total inhaled delivered dose = 35.8 μg / kg body weight) generated at 1V and applied for 15min produced cough in 4 of 6 guinea pigs. The average number of coughs for this exposure was 24 ± 12 times. Cough was observed in 3 of 4 guinea pigs at 0.75 volts and applied for 15min (total inhaled delivered dose = 12.8 μg / kg body weight), and the average cough count was 19 ± 7 times. Cough was not observed in 4 of 4 guinea pigs at 0.5V and applied for 15min (total inhaled delivered dose = 2.3 μg / kg body weight). A mannitol vehicle aerosol generated and applied for 15min at a setting of 1V did not produce cough in 4 of 4 guinea pigs (Table 38). Ventilation changes were inconsistent in the case of formulation D or mannitol vehicle, and the small increase in Penh observed in the case of this drug compared to the mannitol vehicle did not reach statistical significance.

[0309] Nebulized Formulation C (total inhaled delivered dose = 10.2 μg / kg body weight) generated and applied for 15 min at a setting of 1 V produced coughs in 3 of 6 guinea pigs with an average cough count of 10 ± 5 times. Formulation C did not induce coughs in 4 of 4 guinea pigs in either group at settings of 0.75 V (total inhaled delivered dose = 4.7 μg / kg body weight) and 0.5 V (total inhaled delivered dose = 1.5 μg / kg body weight) for 15 min of exposure. Trehalose vehicle applied at a voltage of 1 V for 15 min did not produce coughs in 4 of 4 guinea pigs (Table 38). There was no consistent change in ventilation or Penh with Formulation C or Trehalose vehicle.

[0310] In both Formulation D and Formulation C, lung Treprostinil palmitate equivalent concentrations increased as a function of inhaled drug dose, and Formulation D had approximately 3-fold higher Treprostinil palmitate equivalent levels in the lung compared to Formulation C (Table 38). The two formulations did not differ in percentage of drug deposition in the upper airway tissues of the larynx, trachea, and carina + bronchi, as most of the inhaled drug was deposited in the lungs (data not shown). No nasal tissue was collected in this study.

[0311] The results of this study showed that coughing occurred with both formulations and was observed at an inhalation threshold dose of 12.8 μg / kg for Formulation D and 10.2 μg / kg for Formulation C. These respective doses were 10 and 8 times, respectively, the inhalation TRE threshold dose that induced coughing in guinea pigs (i.e., 1.23 μg / kg). The first cough occurred between 17 and 35 minutes after exposure to either Formulation D or Formulation C, which was later than the time of coughing with aerosolized TRE, which occurred within the first 10 min of exposure.

[0312] The concentration of Treprostinil palmitate equivalents in the lung was approximately three times higher with Formulation D (Table 38), and there was no difference between the two formulations in the percentage of drug deposited in the upper airways of the larynx, trachea, and carina plus bronchi, as most of the drug was deposited in the lung (data not shown). The concentration of TRE in plasma was two- to three-fold higher with Formulation D compared to Formulation C (Table 38). Example 7 - Effect of Treprostinil Palmitate Dry Powder Formulation on the Treatment of 8 Weeks of Sugen Hypoxia (SuHx)-Induced Pulmonary Artery Disease Evaluation of the effect of sera in a rat model of arterial hypertension

[0313] The Sugen hypoxia (SuHx)-induced rat PAH model is a well-documented model. The model replicates many of the pathologies seen in clinical disease. In this example, Treprostinil palmitate dry powder Formulation D (described in Examples 2 and 4; see formulation composition in Table 35A) and inhaled Treprostinil (TRE), intravenous Treprostinil (TRE), and oral Selexipag were evaluated for their effects on an 8-week SuHx-induced rat pulmonary hypertension (PAH) model, including pulmonary artery pressure (PAP) and other cardiovascular parameters, right ventricular hypertrophy, lung and heart histopathology, and biomarkers associated with PAH. Study Groups, Test Articles and Vehicles and Administration

[0314] Male Sprague Dawley rats weighing between 200 g and 250 g were randomized into study groups based on body weight to ensure that the weight range was evenly distributed among the groups. Table 39 summarizes the study groups and the treatment each study group received. 1. Group 1 - Normoxia Control Group

[0315] The normoxic control group (Group 1) received one subcutaneous injection of 2 mL / kg 100% DMSO (vehicle for sugen) and no treatment. 2. Group 2 - Treatment group using vehicle for Treprostinil palmitate dry powder formulation D

[0316] The vehicle for Treprostinil palmitate dry powder formulation D has a target composition of 70 wt% mannitol and 30 wt% leucine. The rats in the vehicle-treated group (Group 2) were weighed and placed in the cup of the Vilnius aerosol generator (VAG) (loaded with 170 mg of vehicle) and administered in a nose cone chamber at 1 volt, once a day, until all the material was aerosolized. The duration of aerosolization was measured. In parallel with Groups 3 and 4, the vehicle treatment of this group was carried out for 35 days. 3. Groups 3 and 4 - Treatment groups using Treprostinil Palmitate Dry Powder Formulation D

[0317] For Groups 3 and 4, Treprostinil palmitate dry powder formulation D was weighed and placed in the cup of the VAG (90 mg was used and it was produced at 0.5 volts (V) for a target dose of 57 μg / kg; 170 mg was used and it was produced at 1 V for a target dose of 138 μg / kg). The dry powder formulation was given once a day for 35 days, and for each application, the VAG remained on until all the material was aerosolized. The duration of aerosolization was measured. The dry powder formulation was stored at 4°C ± 2°C. 4. Group 5 - Treatment group using aerosol vehicle for inhalation of Treprostinil (TRE)

[0318] The aerosol vehicle for inhaled Treprostinil (TRE) was phosphate buffered saline (PBS). In parallel with Group 6, rats in this aerosol vehicle treatment group (Group 5) received aerosolized PBS in a nose cone chamber 4 times daily over a 12-hour period for 35 days. 5. Group 6 - Treatment with aerosolized Treprostinil (TRE)

[0319] The aerosolized Treprostinil (TRE) solution contained 0.5 mM TRE in PBS at pH 7.4. The solution was stored at 4°C ± 2°C and the expiration date was set at 7 days after preparation. The solution was used to deliver a target dose of 110 μg / kg TRE and was administered by inhalation to rats in Group 6 4 times a day over a 12-hour period for 35 days. 6. Group 7 - Treatment group using vehicle for intravenous treprostinil via continuous infusion

[0320] The vehicle for intravenous Treprostinil was an aqueous solution containing 3.0 mg / mL m-cresol, 5.3 mg / mL NaCl, and 6.3 mg / mL sodium citrate at a pH of 6.0-7.2. Each rat in this group (Group 7) was implanted with an osmotic pump filled with vehicle and continuously infused at the infusion rate specified below for Group 8. In parallel with Group 8, the continuous vehicle infusion for Group 7 continued for 35 days. 7. Group 8 - Treatment with intravenous treprostinil (TRE) via continuous infusion

[0321] Two solutions were prepared for intravenous administration of TRE in Group 8. They differed only in the concentration of TRE. Specifically, the first solution was an aqueous solution containing 8.75 mg / mL TRE, 3.0 mg / mL meta-cresol, 5.3 mg / mL NaCl and 6.3 mg / mL sodium citrate at a pH of 6.0-7.2. The second solution was an aqueous solution containing 10.7 mg / mL TRE, 3.0 mg / mL meta-cresol, 5.3 mg / mL NaCl and 6.3 mg / mL sodium citrate at a pH of 6.0-7.2. Each rat in this group (Group 8) received intravenous infusion of TRE using an implanted osmotic pump (ALZET pump). Each ALZET pump was first filled with 2 mL of the first solution containing 8.75 mg / mL TRE, which was sufficient to achieve continuous infusion (based on an infusion rate of 2.5 μL / h) for 450 g rats over a 28-day period and reach a target TRE dose of 810 ng / kg / min. On day 19 of infusion (day 40 of the entire study), the ALZET pump was replaced and filled with 2 mL of the second solution containing 10.7 mg / mL TRE based on the rat weight increase to approximately 550 g. The derivation of TRE concentrations in the first and second solutions is shown below. Continuous IV TRE infusion continued for 35 days.

[0322] Derivation of TRE concentration (8.75 mg / mL) in the first solution: o Assuming rat weight is 450g o Infused TRE = 810 ng / kg / min; = 364.5 ng / min; = 21.87 μg / h; = 524.88 μg / day; = 18.37 mg / 35 days o AlZET infusion rate = 2.5 μL / h; = 60 μL / day; = 2100 μL / 35 days; = 2.1 mL / 35 days o TRE concentration = 18.37 mg / 2.1 mL = 8.75 mg / mL

[0323] Derivation of TRE concentration (10.7 mg / mL) in the second solution o Assuming rat weight is 550g o Infused TRE = 810 ng / kg / min; = 445.5 ng / min; = 26.73 μg / h; = 641.52 μg / day; = 22.45mg / 35 days o ALZET infusion rate = 2.5 μL / h; = 60 μL / day; = 2100 μL / 35 days; = 2.1 mL / 35 days o TRE concentration = 22.45 mg / 2.1 mL = 10.7 mg / mL 8. Group 9 - Treatment group using vehicle for selexipag via oral administration

[0324] The vehicle for selexipag was an aqueous solution at pH 7.5-8.0 containing 0.5% (w / v) methylcellulose. In parallel with Group 10, rats in this group were dosed with vehicle by oral gavage twice a day for 35 days. 9. Group 10 - Treatment group using selexipag via oral administration

[0325] A selexipag solution at pH 7.5 containing 3.0 mg / mL selexipag in 0.5% (w / v) methylcellulose was prepared. The solution was stored at room temperature and the shelf life was set at 7 days after preparation. The solution was administered to the group of rats (Group 10) by oral gavage twice a day at a volume of 10 mL / kg at each administration, with a target dose of 30 mg / kg. Selexipag treatment lasted for 35 days. Study Design

[0326] The study design is summarized in Table 39 and its details are as follows. 1. Induction of PAH

[0327] On day 21, rats were randomized into treatment groups according to their body weight as described above.

[0328] On day 0, a 10 mg / mL solution of sugen in DMSO was prepared and rats from groups 2 to 10 (see Table 39) received a single subcutaneous injection of sugen (20 mg / kg in a 2 mL / kg volume) solution and returned to their cages. Also on day 0, rats in group 1 received a single subcutaneous injection of 2 mL / kg of 100% DMSO (vehicle for sugen) and returned to their respective cages.

[0329] The rats in groups 2-10 were placed in cages with controlled atmosphere conditioned to receive an FiO2 equivalent to 0.10 (10%) using a mixture of nitrogen and ambient air controlled by a ventilated cage system. They were maintained under these hypoxic conditions for 21 days. While hypoxic, the cages were cleaned and changed once a week, exposing the rats to ambient oxygen levels for less than 10 minutes. They were exposed to ambient oxygen levels from day 22 to day 56. The rats in group 1 were maintained in cages exposed to ambient oxygen (normoxia) levels for 56 days. The rats were observed daily for any changes in behavior and general health.

[0330] Treatment with test article or vehicle was administered from day 22 to day 56. Food and water were given ad libitum. The behavior and general health of the rats were observed daily. Body weights were obtained once a week. 2. Echocardiography

[0331] All rats were monitored for disease progression by echocardiography on day 0, day 21 (before treatment), and on the day of surgery (day 56). 3. Blood and Lung PK Sampling

[0332] Venous blood (0.5 ml, anticoagulated with EDTA) was sampled from all rats on day 23 (just before the second dose), day 38 (just before the next dose), and day 57 (24 hours after the last dose). Blood was collected from the saphenous vein in rats with ALZET pumps, and from the jugular vein in all other rats. Whole blood was centrifuged to obtain plasma, which was stored frozen at -80°C for analysis. 4. Dry powder inhalation (Group 2-4)

[0333] During the treatment period, each rat was placed in a nose cone restraint chamber, which was connected to a 12-port nasal inhalation chamber (CH Technologies). Treprostinil palmitate dry powder formulation D or its vehicle was delivered using VAG. Airflow was introduced into the VAG at a flow rate of 7L / min and connected to the nasal inhalation chamber. For the 3rd group treated with a lower dose of Treprostinil palmitate dry powder formulation D, 90mg Treprostinil palmitate dry powder formulation D was weighed and loaded into the VAG depolymerizer. The VAG was set to a voltage of 0.5V, which corresponds to a powder concentration of 0.5mg / L (about 7μg / L Treprostinil palmitate). For the 4th group treated with a higher dose of Treprostinil palmitate dry powder formulation D, 170mg Treprostinil palmitate dry powder formulation D was weighed and loaded into the VAG depolymerizer and delivered at a voltage of 1V, which corresponds to a powder concentration of 1.0mg / L (about 14.7μg / L Treprostinil palmitate). The powder aerosol output concentration is continuously monitored by a portable aerosol monitor (CasellaMicroDust Pro). The accurate delivery time is recorded. The dry powder remaining in the VAG deagglomerator is weighed to calculate the actual amount of the test dry powder atomized. 5min after the start of atomization, a glass fiber filter connected to a vacuum source at a vacuum flow rate of 0.5L / min is placed on one of the exposed ports to collect the aerosol from the chamber on the filter for a period of 5 minutes. All filter samples are kept at 4°C until analysis.

[0334] Rats from Group 2 received 170 mg of vehicle for Treprostinil palmitate dry powder Formulation D administered at a 1.0 V setting.

[0335] In this study, two different inhalation towers and multiple sets of VAGs and lasers were used, one for the dry powder vehicle and the other for the Treprostinil palmitate dry powder formulation D.

[0336] After the remaining powder was taken out from the deagglomerator of the VAG, all parts of the VAG were purged with dry air. The tower was purged with dry air between the 3rd (low dose) and 4th (high dose) administrations, and was cleaned with 0.5% aqueous solution of sodium dodecyl sulfate (SDS), tap water, and distilled water after the 4th administration. 5. Aerosol inhalation (Groups 5 and 6)

[0337] Treprostinil and its vehicle PBS were administered using an aerosolizer and controller (Aeroneb Pro) from Aerogen, which was manufactured for delivery of mass mean aerosol diameters (MMAD) between 2.5 and 4 μm and flow rates in the range of 0.2-0.4 mL / min. During the treatment period, each rat was placed in a nose cone restraint chamber connected to a 12-port nasal inhalation chamber (CH Technologies). The volume of the solution to be aerosolized was 6 mL and the airflow was 6 L / min, and the concentration of Treprostinil was 0.5 mM. A glass fiber filter was placed on one of the exposed ports and connected to a vacuum source at a vacuum flow rate of 0.5 L / min for a period of 5 minutes, starting 5 min after aerosolization began and ending 10 min later.

[0338] Two inhalation towers and two separate sets of nose cones were used; one for Group 5 receiving PBS and one for Group 6 receiving TRE.

[0339] The cleaning of the aerosolizer was performed by flowing a 0.5% SDS aqueous solution, tap water, and distilled water sequentially through the aerosolizer, and aerosolizing PBS between each use to wash out any residual drug from the drug cup and through the orifice plate. The aerosol tower and other materials used in the aerosolization process were also cleaned with the above-mentioned agent once a day. In addition, the aerosol tower and other materials used in the aerosolization process were cleaned with the above-mentioned agent after each chamber administration. 6. IV continuous infusion (Groups 7 and 8)

[0340] Rats from Group 8 were anesthetized with isoflurane 2% and medical grade air. An incision was made in the back of each rat to place an ALZET pump filled with a first solution containing 8.75 mg / mL TRE. A catheter was implanted in the jugular vein and connected to the ALZET pump. For continuous infusion over a period of 35 days, on the 19th day of infusion, the catheter was temporarily clamped and the ALZET pump was replaced with a new pump filled with a second solution containing 10.7 mg / mL TRE. Each rat from Group 7 was implanted with an ALZET pump filled with vehicle. 7. Oral administration (Groups 9 and 10)

[0341] Rats from Group 10 received the reference compound Selexipag by oral gavage from Day 22 to Day 56 (35 days), twice a day. Care was taken to maintain a uniform Selexipag suspension by continuous stirring while the dose was introduced into the gavage syringe, filling one gavage syringe at a time and applying the dose before filling the next syringe. The dose was given by oral gavage by administering 10 mL / kg body weight each time. Rats from Group 9 were administered with a vehicle by oral gavage from Day 22 to Day 56 (35 days), twice a day. Surgical Instrumentation and Measurement of Hemodynamic and Functional Parameters in Efficacy Study Rats

[0342] 1. On the selected surgical day, 24 hours after the last dose, rats were anesthetized with a mixture of 2%-2.5% isoflurane USP (Abbot Laboratories) in oxygen and placed on a heating pad to maintain body temperature.

[0343] 2. The rats were tracheotomized and immediately ventilated via a positive pressure rodent ventilator set at ≈10 ml / kg body weight and a frequency of 90 breaths / min.

[0344] 3. A cannula connected to a pressure transducer was inserted into the left femoral artery to measure systemic arterial blood pressure (SAP).

[0345] 4. The heart was exposed through sternotomy and a 20GA 1.16 in Insyte was introduced into the right ventricle and quickly hooked to a saline filled PE-50 catheter connected to the transducer.

[0346] 5. After the right ventricular pressure is recorded for a few seconds, the Insyte is advanced further into the pulmonary artery to allow PAP to be recorded for an additional 60 seconds.

[0347] 6. Hemodynamic parameters were recorded continuously for the duration of the procedure or until the PAP signal was lost.

[0348] 7. Following hemodynamic monitoring, blood was obtained by cardiac puncture for biomarker analysis (described below).

[0349] 8. After collecting the blood samples, the chest cavity was further opened to expose the lungs. The muscles over the trachea were dissected away to remove the lungs and heart. The harvested tissues were rinsed with PBS to remove any excess blood before weighing.

[0350] 9. The right lung was knotted and immediately collected for drug concentration and biomarker analysis by separating the four lobes, which were weighed and frozen in liquid nitrogen and stored at -80°C.

[0351] 10. For cardiac histology and casting, the procedure was as follows: 1) 5 of the 11 rats in each of groups 2-10 were used to assess cardiac histology and biochemical parameters and were therefore processed as described in point 11 below; 2) The other 6 rats in groups 2-10 were used to determine the Fulton index and were processed as described in point 12. After the data for the Fulton index were collected, the cardiac tissues were stored at -80°C for biomarker analysis.

[0352] 11. For histology, the left lung was flushed with 0.9% NaCl. The left lung was inflated using a 10 mL syringe filled with fixative, 10% neutral buffered formalin (NBF), and an attached blunt needle (23 g). The needle tip was inserted into the trachea and fixed with a tied suture, while the other syringe was tied to the pulmonary artery. The lung was gently inflated under physiological pressure until the lung was fully, evenly and continuously expanded (fixative was not allowed to seep out of the lung surface). This provides optimal vascular and airway expansion without excessive tissue destruction. The needle was then removed, the suture was tied around the trachea, and the lung was immersed in 10% NBF at a tissue to fixative ratio of 1:20. The heart was rinsed in PBS and then immersed in 10% NBF at a tissue to fixative ratio of 1:20. The tissue was stored in formalin for 24-48h. The left lung and heart were then cut and transferred to 70% ethanol.

[0353] All fixed tissues were embedded, sectioned and stained. Lung sections were stained with hematoxylin and eosin (H&E) for morphological determination or von Willebrand factor (VWF) staining for endothelial cells. Heart sections were stained with H&E and stained with Sirius red or trichrome staining for collagen fiber visualization and quantification.

[0354] 12. As part of the Fulton Index, the heart was dissected to separate the right ventricle from the left ventricle with a septum and then weighed separately. After collecting the data for the Fulton Index, the heart tissue was stored at -80°C for biomarker analysis. Acquisition and analysis of experimental data

[0355] Experimental traces were analyzed by Clampfit software from Axon Instruments.

[0356] PAP recorded continuously for at least 1 min or until signal loss was used to determine mean, diastolic, and systolic pulmonary pressures.

[0357] Continuously recorded systemic arterial pressure (SAP) was used to determine mean arterial pressure, diastolic arterial pressure, and systolic arterial pressure.

[0358] At the end of the recording of hemodynamic parameters, the right and left ventricles including the diaphragm and the lung lobes were removed to determine the wet weight.

[0359] The following hemodynamic and cardiac function parameters were quantified by appropriate statistical analysis. Mean arterial pressure Mean arterial pulmonary pressure Diastolic pulmonary pressure Systolic pulmonary pressure Systolic right ventricular pressure Saturation (SO2) Weight gain Lung weight Fulton Index Heart rate Pulse pressure

[0360] In addition, molecules indicative of cardiac biochemistry were examined, including biomarkers of oxidative stress, namely collagen (Sircol assay) and hydroxyproline content, uric acid, natriuretic peptides (BNP, NT-proBNP (biomarkers of myocardial stress)), endothelin-1 (heart failure), angiopoietin (neovascularization), von Willebrand factor (endothelial cells), interleukin-6 (biomarker of heart attack, stroke), Toll receptor C (biomarker of heart disease), plasma cytokines, atrial natriuretic peptide ANP (biomarker of stroke, coronary artery disease, myocardial infarction and heart failure), troponin T / I (biomarker of cardiac ischemia) and CPK-MB (cardiac biomarker of myocardial infarction).

[0361] Genes associated with PAH, such as bone morphogenetic protein type 2 (BMPR1), BMP-9, ABCC8, TBX4, ACVRL, SMAD 4 / 9, KCNA5, and TET2, were also studied in this example. In addition, in the heart and lungs, genes such as type 1 collagen alpha 1 (COL1A1), type 1 collagen alpha 2 (COL1A2), and type 3 collagen alpha 1 (COL3A1) are associated with the formation and secretion of collagen. P4HA1 is a key enzyme in collagen biosynthesis. ACTG2 is a gene associated with myofibroblast differentiation. Changes in the expression of those genes were also studied.

[0362] It is expected that Treprostinil palmitate dry powder Formulation D will improve pathophysiology and histopathology in the pulmonary vasculature and heart of Su / Hx-challenged rats. *********

[0363] Although the invention has been described with reference to specific embodiments of the invention, it will be appreciated by those skilled in the art that various changes may be made and equivalents may be substituted without departing from the true spirit and scope of the invention. In addition, many modifications may be made to adopt specific circumstances, materials, compositions of matter, processes, process steps or steps to achieve the objective spirit and scope of the invention. It is contemplated that all such modifications are within the scope of the appended claims.

[0364] The patents, patent applications, patent application publications, journal articles, and protocols cited herein are incorporated by reference in their entirety for any purpose.

[0365] This application also covers implementations of: Item 1. A dry powder composition, comprising: (a) from about 0.1 wt % to about 3 wt % of a compound of formula (I): or an enantiomer, a diastereomer or a pharmaceutically acceptable salt thereof, wherein R 1 is tetradecyl, pentadecyl, hexadecyl, heptadecyl or octadecyl, (b) about 0.01 wt % to about 3 wt % of DSPE-PEG2000, (c) from about 10 wt % to about 50 wt % leucine, and The rest is (d) a sugar selected from trehalose and mannitol, The total of (a), (b), (c) and (d) is 100 wt %. Item 2. The dry powder composition according to Item 1, wherein (a) is a compound of formula (I) or a pharmaceutically acceptable salt thereof. Item 3. The dry powder composition according to Item 1 or 2, wherein (a) is a compound of formula (I). Item 4. A dry powder composition according to any one of Items 1 to 3, wherein R 1 It is tetradecyl. Item 5. A dry powder composition according to Item 4, wherein R 1 It is a linear tetradecyl group. Item 6. A dry powder composition according to any one of Items 1 to 3, wherein R 1 It is a pentadecyl group. Item 7. A dry powder composition according to Item 6, wherein R 1 It is a linear pentadecyl group. Item 8. A dry powder composition according to any one of Items 1 to 3, wherein R 1 It is heptadecanyl. Item 9. A dry powder composition according to Item 8, wherein R1 It is a linear heptadecanyl group. Item 10. A dry powder composition according to any one of Items 1 to 3, wherein R 1 It is octadecyl. Item 11. A dry powder composition according to Item 10, wherein R 1 It is a linear octadecyl group. Item 12. A dry powder composition according to any one of Items 1 to 3, wherein R 1 It is hexadecyl. Item 13. A dry powder composition according to Item 12, wherein R 1 It is a linear hexadecyl group. Item 14. The dry powder composition of any one of Items 1-13, wherein the DSPE-PEG2000 is present at about 0.03 wt % to about 2.1 wt % of the total weight of the dry powder composition. Item 15. The dry powder composition of any one of Items 1-14, wherein the DSPE-PEG2000 is present at about 0.05 wt % to about 1.5 wt % of the total weight of the dry powder composition. Item 16. A dry powder composition according to any one of Items 1-13, wherein the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in an amount of about 0.5 wt % to about 2 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in an amount of about 0.05 wt % to about 2 wt % of the total weight of the dry powder composition. Item 17. A dry powder composition according to Item 16, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is present in an amount of about 0.5 wt % to about 2 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in an amount of about 0.05 wt % to about 2 wt % of the total weight of the dry powder composition. Item 18. A dry powder composition according to Item 16 or 17, wherein the compound of formula (I) is present at about 0.5 wt % to about 2 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.05 wt % to about 2 wt % of the total weight of the dry powder composition. Item 19. The dry powder composition of any one of Items 16-18, wherein the DSPE-PEG2000 is present at about 0.15 wt % to about 1.4 wt % of the total weight of the dry powder composition. Item 20. The dry powder composition of any one of Items 16-19, wherein the DSPE-PEG2000 is present at about 0.25 wt % to about 1 wt % of the total weight of the dry powder composition. Item 21. A dry powder composition according to any one of Items 1-13, wherein the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in an amount of about 1 wt % to about 2 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in an amount of about 0.1 wt % to about 2 wt % of the total weight of the dry powder composition. Item 22. A dry powder composition according to Item 21, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is present in an amount of about 1 wt % to about 2 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in an amount of about 0.1 wt % to about 2 wt % of the total weight of the dry powder composition. Item 23. A dry powder composition according to Item 21 or 22, wherein the compound of formula (I) is present at about 1 wt% to about 2 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.1 wt% to about 2 wt% of the total weight of the dry powder composition. Item 24. The dry powder composition of any one of Items 21-23, wherein the DSPE-PEG2000 is present at about 0.3 wt % to about 1.4 wt % of the total weight of the dry powder composition. Item 25. The dry powder composition of any one of Items 21-24, wherein the DSPE-PEG2000 is present at about 0.5 wt % to about 1 wt % of the total weight of the dry powder composition. Item 26. A dry powder composition according to any one of Items 1-13, wherein the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.2 wt% to about 1.8 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.12 wt% to about 1.8 wt% of the total weight of the dry powder composition. Item 27. A dry powder composition according to Item 26, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is present at about 1.2 wt% to about 1.8 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.12 wt% to about 1.8 wt% of the total weight of the dry powder composition. Item 28. A dry powder composition according to Item 26 or 27, wherein the compound of formula (I) is present at about 1.2 wt% to about 1.8 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.12 wt% to about 1.8 wt% of the total weight of the dry powder composition. Item 29. The dry powder composition of any one of Items 26-28, wherein the DSPE-PEG2000 is present at about 0.36 wt % to about 1.26 wt % of the total weight of the dry powder composition. Item 30. The dry powder composition of any one of Items 26-29, wherein the DSPE-PEG2000 is present at about 0.6 wt % to about 0.9 wt % of the total weight of the dry powder composition. Item 31. A dry powder composition according to any one of Items 1-13, wherein the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present in an amount of about 1 wt % to about 1.5 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in an amount of about 0.1 wt % to about 1.5 wt % of the total weight of the dry powder composition. Item 32. A dry powder composition according to Item 31, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is present in an amount of about 1 wt % to about 1.5 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present in an amount of about 0.1 wt % to about 1.5 wt % of the total weight of the dry powder composition. Item 33. A dry powder composition according to Item 31 or 32, wherein the compound of formula (I) is present at about 1 wt% to about 1.5 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.1 wt% to about 1.5 wt% of the total weight of the dry powder composition. Item 34. The dry powder composition of any one of Items 31-33, wherein the DSPE-PEG2000 is present at about 0.3 wt % to about 1.05 wt % of the total weight of the dry powder composition. Item 35. The dry powder composition of any one of Items 31-34, wherein the DSPE-PEG2000 is present at about 0.5 wt % to about 0.75 wt % of the total weight of the dry powder composition. Item 36. A dry powder composition according to any one of Items 1-13, wherein the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1.4 wt% to about 1.6 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.14 wt% to about 1.6 wt% of the total weight of the dry powder composition. Item 37. A dry powder composition according to Item 36, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is present at about 1.4 wt % to about 1.6 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.14 wt % to about 1.6 wt % of the total weight of the dry powder composition. Item 38. A dry powder composition according to Item 36 or 37, wherein the compound of formula (I) is present at about 1.4 wt% to about 1.6 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.14 wt% to about 1.6 wt% of the total weight of the dry powder composition. Item 39. The dry powder composition of any one of Items 36-38, wherein the DSPE-PEG2000 is present at about 0.42 wt % to about 1.12 wt % of the total weight of the dry powder composition. Item 40. The dry powder composition of any one of Items 36-39, wherein the DSPE-PEG2000 is present at about 0.7 wt % to about 0.8 wt % of the total weight of the dry powder composition. Item 41. A dry powder composition according to any one of Items 1-13, wherein the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at about 1 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.1 wt% to about 1 wt% of the total weight of the dry powder composition. Item 42. A dry powder composition according to Item 41, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is present at about 1 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.1 wt % to about 1 wt % of the total weight of the dry powder composition. Item 43. A dry powder composition according to Item 41 or 42, wherein the compound of formula (I) is present at about 1 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.1 wt % to about 1 wt % of the total weight of the dry powder composition. Item 44. The dry powder composition of any one of Items 41-43, wherein the DSPE-PEG2000 is present at about 0.3 wt % to about 0.7 wt % of the total weight of the dry powder composition. Item 45. The dry powder composition of any one of Items 41-44, wherein the DSPE-PEG2000 is present at about 0.5 wt % of the total weight of the dry powder composition. Item 46. A dry powder composition according to any one of Items 1-13, wherein the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is present at approximately 1.5 wt% of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at approximately 0.15 wt% to approximately 1.5 wt% of the total weight of the dry powder composition. Item 47. A dry powder composition according to Item 46, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is present at approximately 1.5 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at approximately 0.15 wt % to approximately 1.5 wt % of the total weight of the dry powder composition. Item 48. A dry powder composition according to Item 46 or 47, wherein the compound of formula (I) is present at about 1.5 wt % of the total weight of the dry powder composition, and the DSPE-PEG2000 is present at about 0.15 wt % to about 1.5 wt % of the total weight of the dry powder composition. Item 49. The dry powder composition of any one of Items 46-48, wherein the DSPE-PEG2000 is present at about 0.45 wt % to about 1.05 wt % of the total weight of the dry powder composition. Item 50. The dry powder composition of any one of Items 46-49, wherein the DSPE-PEG2000 is present at about 0.75 wt % of the total weight of the dry powder composition. Item 51. A dry powder composition, comprising: (a) from about 0.1 wt % to about 3 wt % of a compound of formula (I): or an enantiomer, a diastereomer or a pharmaceutically acceptable salt thereof, wherein R 1 is tetradecyl, pentadecyl, hexadecyl, heptadecyl or octadecyl, (b) DSPE-PEG2000 having a weight ratio of DSPE-PEG2000 to the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt in the range of about 0.1:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt) to about 1:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt), (c) from about 10 wt % to about 50 wt % leucine, and The rest is (d) a sugar selected from trehalose and mannitol, The total of (a), (b), (c) and (d) is 100 wt %. Item 52. A dry powder composition according to Item 51, wherein (a) is a compound of formula (I) or a pharmaceutically acceptable salt thereof. Item 53. A dry powder composition according to Item 51 or 52, wherein (a) is a compound of formula (I). Item 54. A dry powder composition according to any one of Items 51-53, wherein R 1 It is tetradecyl. Item 55. A dry powder composition according to Item 54, wherein R 1 It is a linear tetradecyl group. Item 56. A dry powder composition according to any one of Items 51-53, wherein R 1 It is a pentadecyl group. Item 57. A dry powder composition according to Item 56, wherein R 1 It is a linear pentadecyl group. Item 58. A dry powder composition according to any one of Items 51-53, wherein R 1 It is heptadecanyl. Item 59. A dry powder composition according to Item 58, wherein R 1 It is a linear heptadecanyl group. Item 60. A dry powder composition according to any one of Items 51-53, wherein R 1 It is octadecyl. Item 61. A dry powder composition according to Item 60, wherein R 1 It is a linear octadecyl group. Item 62. A dry powder composition according to any one of Items 51-53, wherein R 1 It is hexadecyl. Item 63. A dry powder composition according to Item 62, wherein R 1 It is a linear hexadecyl group. Item 64. A dry powder composition according to any one of Items 51-63, wherein the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof is present in an amount of about 0.5 wt % to about 2 wt % of the total weight of the dry powder composition. Item 65. A dry powder composition according to any one of Items 51-63, wherein the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof is present in an amount of about 1 wt % to about 2 wt % of the total weight of the dry powder composition. Item 66. A dry powder composition according to any one of Items 51-63, wherein the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof is present in an amount of about 1.2 wt % to about 1.8 wt % of the total weight of the dry powder composition. Item 67. A dry powder composition according to any one of Items 51-63, wherein the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof is present in an amount of about 1 wt % to about 1.5 wt % of the total weight of the dry powder composition. Item 68. A dry powder composition according to any one of Items 51-63, wherein the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof is present in an amount of about 1.4 wt % to about 1.6 wt % of the total weight of the dry powder composition. Item 69. A dry powder composition according to any one of Items 51-63, wherein the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof is present at about 1 wt % of the total weight of the dry powder composition. Item 70. A dry powder composition according to any one of Items 51-63, wherein the compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof is present at about 1.5 wt % of the total weight of the dry powder composition. Item 71. A dry powder composition according to any one of Items 51-70, wherein the weight ratio of the DSPE-PEG2000 to the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is in the range of about 0.3:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt) to about 0.7:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt). Item 72. A dry powder composition according to any one of Items 51-71, wherein the weight ratio of the DSPE-PEG2000 to the compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt is about 0.5:1 (DSPE-PEG2000: compound of formula (I) or its enantiomer, diastereomer or pharmaceutically acceptable salt). Item 73. A dry powder composition according to any one of Items 1-72, wherein the leucine is present at about 15 wt % to about 40 wt % of the total weight of the dry powder composition. Item 74. A dry powder composition according to Item 73, wherein the leucine is present at about 18 wt % to about 33 wt % of the total weight of the dry powder composition. Item 75. A dry powder composition according to Item 74, wherein the leucine is present in an amount of about 20 wt % to about 30 wt % of the total weight of the dry powder composition. Item 76. A dry powder composition according to Item 75, wherein the leucine is present at about 25 wt % to about 30 wt % of the total weight of the dry powder composition. Item 77. A dry powder composition according to Item 76, wherein the leucine is present at about 27 wt % to about 30 wt % of the total weight of the dry powder composition. Item 78. A dry powder composition according to Item 75, wherein the leucine is present at about 20 wt % of the total weight of the dry powder composition. Item 79. A dry powder composition according to Item 75, wherein the leucine is present at about 30 wt % of the total weight of the dry powder composition. Item 80. The dry powder composition of any one of Items 1-79, wherein the sugar is trehalose. Item 81. The dry powder composition of any one of Items 1-79, wherein the sugar is mannitol. Item 82. A dry powder composition according to any one of Items 1-13, comprising (a) approximately 1.5 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) approximately 0.7 wt% of the DSPE-PEG2000, (c) approximately 29.3 wt% of the leucine, and the remainder being (d) trehalose. Item 83. A dry powder composition according to Item 82, comprising (a) about 1.5 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 29.3 wt% of the leucine, and the remainder being (d) trehalose. Item 84. A dry powder composition according to Item 82 or 83, comprising (a) about 1.5 wt% of the compound of formula (I), (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 29.3 wt% of the leucine, and the remainder being (d) trehalose. Item 85. A dry powder composition according to any one of Items 1-13, comprising (a) about 1 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 29.6 wt% of the leucine, and the remainder being (d) trehalose. Item 86. A dry powder composition according to Item 85, comprising (a) about 1 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 29.6 wt% of the leucine, and the remainder being (d) trehalose. Item 87. A dry powder composition according to Item 85 or 86, comprising (a) about 1 wt% of the compound of formula (I), (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 29.6 wt% of the leucine, and the remainder being (d) trehalose. Item 88. A dry powder composition according to any one of Items 1-13, comprising (a) about 1 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 19.7 wt% of the leucine, and the remainder being (d) trehalose. Item 89. A dry powder composition according to Item 88, comprising (a) about 1 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 19.7 wt% of the leucine, and the remainder being (d) trehalose. Item 90. A dry powder composition according to Item 88 or 89, comprising (a) about 1 wt% of the compound of formula (I), (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 19.7 wt% of the leucine, and the remainder being (d) trehalose. Item 91. A dry powder composition according to any one of Items 1-13, comprising (a) about 1.5 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 19.6 wt% of the leucine, and the remainder being (d) trehalose. Item 92. A dry powder composition according to Item 91, comprising (a) about 1.5 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 19.6 wt% of the leucine, and the remainder being (d) trehalose. Item 93. A dry powder composition according to Item 91 or 92, comprising (a) about 1.5 wt% of the compound of formula (I), (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 19.6 wt% of the leucine, and the remainder being (d) trehalose. Item 94. A dry powder composition according to any one of Items 1-13, comprising (a) about 1.5 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 29.3 wt% of the leucine, and the remainder being (d) mannitol. Item 95. A dry powder composition according to Item 94, comprising (a) about 1.5 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 29.3 wt% of the leucine, and the remainder being (d) mannitol. Item 96. A dry powder composition according to Item 94 or 95, comprising (a) about 1.5 wt% of the compound of formula (I), (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 29.3 wt% of the leucine, and the remainder being (d) mannitol. Item 97. A dry powder composition according to any one of Items 1-13, comprising (a) approximately 1.5 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) approximately 0.75 wt% of the DSPE-PEG2000, (c) approximately 29.30 wt% of the leucine, and (d) approximately 68.45 wt% of the mannitol. Item 98. A dry powder composition according to Item 97, comprising (a) approximately 1.5 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) approximately 0.75 wt% of the DSPE-PEG2000, (c) approximately 29.30 wt% of the leucine, and (d) approximately 68.45 wt% of the mannitol. Item 99. A dry powder composition according to Item 97 or 98, comprising (a) approximately 1.5 wt% of the compound of formula (I), (b) approximately 0.75 wt% of the DSPE-PEG2000, (c) approximately 29.30 wt% of the leucine, and (d) approximately 68.45 wt% of the mannitol. Item 100. A dry powder composition according to any one of Items 1-13, comprising (a) about 1 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 29.6 wt% of the leucine, and the remainder being (d) mannitol. Item 101. A dry powder composition according to Item 100, comprising (a) about 1 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 29.6 wt% of the leucine, and the remainder being (d) mannitol. Item 102. A dry powder composition according to Item 100 or 101, comprising (a) about 1 wt% of the compound of formula (I), (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 29.6 wt% of the leucine, and the remainder being (d) mannitol. Item 103. A dry powder composition according to any one of Items 1-13, comprising (a) about 1.5 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 19.6 wt% of the leucine, and the remainder being (d) mannitol. Item 104. A dry powder composition according to Item 103, comprising (a) about 1.5 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 19.6 wt% of the leucine, and the remainder being (d) mannitol. Item 105. A dry powder composition according to Item 103 or 104, comprising (a) about 1.5 wt% of the compound of formula (I), (b) about 0.7 wt% of the DSPE-PEG2000, (c) about 19.6 wt% of the leucine, and the remainder being (d) mannitol. Item 106. A dry powder composition according to any one of Items 1-13, comprising (a) about 1 wt% of a compound of formula (I) or an enantiomer, diastereomer or pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 19.7 wt% of the leucine, and the remainder being (d) mannitol. Item 107. A dry powder composition according to Item 106, comprising (a) about 1 wt% of the compound of formula (I) or a pharmaceutically acceptable salt thereof, (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 19.7 wt% of the leucine, and the remainder being (d) mannitol. Item 108. A dry powder composition according to Item 106 or 107, comprising (a) about 1 wt% of the compound of formula (I), (b) about 0.5 wt% of the DSPE-PEG2000, (c) about 19.7 wt% of the leucine, and the remainder being (d) mannitol. Item 109. A dry powder composition according to any one of items 1-108, wherein the dry powder composition is in the form of an aerosol comprising particles having a mass median aerodynamic diameter (MMAD) of about 1 μm to about 3 μm as measured by a Next Generation Impactor (NGI). Item 110. The dry powder composition of any one of items 1-109, wherein the dry powder composition is in the form of an aerosol comprising particles having an MMAD of about 1.3 μm to about 2.0 μm as measured by NGI. Item 111. The dry powder composition of any one of items 81 and 94-108, wherein the dry powder composition is in the form of an aerosol comprising particles having an MMAD of about 1.7 μm to about 2.7 μm as measured by NGI. Item 112. The dry powder composition of any one of Items 1-111, wherein the dry powder composition is in the form of an aerosol comprising particles having a fine particle fraction of about 30% to about 60% as measured by NGI. Item 113. A method for treating pulmonary hypertension in a patient in need thereof, the method comprising administering an effective amount of a dry powder composition according to any one of Items 1-112 to the lungs of the patient by inhalation via a dry powder inhaler. Item 114. A method according to Item 113, wherein the pulmonary hypertension is pulmonary arterial hypertension. Item 115. The method of Item 114, wherein the pulmonary hypertension is Class I pulmonary hypertension as characterized by the New York Heart Association (NYHA). Item 116. A method according to Item 114, wherein the pulmonary hypertension is Class II pulmonary hypertension as characterized by NYHA. Item 117. A method according to Item 114, wherein the pulmonary hypertension is Class III pulmonary hypertension as characterized by NYHA. Item 118. A method according to Item 114, wherein the pulmonary hypertension is Class IV pulmonary hypertension as characterized by NYHA. Item 119. A method according to Item 113, wherein the pulmonary hypertension is Group 1 pulmonary hypertension as characterized by the World Health Organization (WHO). Item 120. A method according to Item 113, wherein the pulmonary hypertension is Group 2 pulmonary hypertension as characterized by WHO. Item 121. A method according to Item 113, wherein the pulmonary hypertension is Group 3 pulmonary hypertension as characterized by WHO. Item 122. A method according to Item 113, wherein the pulmonary hypertension is Group 4 pulmonary hypertension as characterized by WHO. Item 123. A method according to Item 113, wherein the pulmonary hypertension is Group 5 pulmonary hypertension as characterized by WHO. Item 124. A method for treating portopulmonary hypertension or pulmonary fibrosis in a patient in need thereof, the method comprising administering an effective amount of a dry powder composition according to any one of Items 1-112 to the lungs of the patient by inhalation via a dry powder inhaler. Item 125. The method according to any one of Items 113-124, wherein the administration is carried out in the form of once-a-day, twice-a-day or three-a-day dosing. Item 126. The method of any one of Items 113-125, wherein the administering comprises aerosolizing the dry powder composition and administering the aerosolized dry powder composition to the patient's lungs via inhalation. Item 127. The method of Item 126, wherein the aerosolized dry powder composition comprises particles having an MMAD of about 1 μm to about 3 μm as measured by NGI. Item 128. The method of Item 126 or 127, wherein the aerosolized dry powder composition comprises particles having a fine particle fraction of about 30% to about 60% as measured by NGI. Item 129. A system for treating pulmonary hypertension, portopulmonary hypertension, or pulmonary fibrosis, the system comprising: A dry powder composition according to any one of items 1 to 112, and Dry powder inhaler (DPI). Item 130. The system of Item 129, wherein the DPI is a single-dose or multi-dose inhaler. Item 131. A system according to Item 129, wherein the DPI is pre-metered or device-metered.

Claims

1. A dry powder composition, comprising: (a) from about 0.1 wt % to about 3 wt % of a compound of formula (I): or an enantiomer, a diastereomer or a pharmaceutically acceptable salt thereof, wherein R 1 is tetradecyl, pentadecyl, hexadecyl, heptadecyl or octadecyl, (b) about 0.01 wt % to about 3 wt % of DSPE-PEG2000, (c) from about 10 wt % to about 50 wt % leucine, and The rest is (d) a sugar selected from trehalose and mannitol, The total of (a), (b), (c) and (d) is 100 wt %.

2. The dry powder composition according to claim 1, wherein (a) is a compound of formula (I) or a pharmaceutically acceptable salt thereof.

3. A dry powder composition according to claim 1 or 2, wherein (a) is a compound of formula (I).

4. The dry powder composition according to any one of claims 1 to 3, wherein R 1 It is tetradecyl.

5. The dry powder composition according to claim 4, wherein R 1 It is a linear tetradecyl group.

6. A dry powder composition according to any one of claims 1 to 3, wherein R 1 It is a pentadecyl group.

7. The dry powder composition according to claim 6, wherein R 1 It is a linear pentadecyl group.

8. A dry powder composition according to any one of claims 1 to 3, wherein R 1 It is heptadecanyl.

9. The dry powder composition according to claim 8, wherein R 1 It is a linear heptadecanyl group.

10. The dry powder composition according to any one of claims 1 to 3, wherein R 1 It is octadecyl.

Images