Dry powder treprostinil for treatment of pulmonary hypertension

Abstract

To provide a dry powder inhalation treatment for pulmonary arterial hypertension, a method of treating a patient suffering from pulmonary arterial hypertension, a dry powder inhalation composition for treatment, and a method of producing particles for delivering a dry powder to the lungs of a patient with pulmonary arterial hypertension.SOLUTION: The dry powder inhalation treatment for pulmonary arterial hypertension includes a dose of dry particles comprising more than 25 micrograms of treprostinil enclosed in a capsule. The dry particles may include treprostinil, a wetting agent, a hydrophobicity modifying agent, a pH modifying agent, and a buffer. A method of treating a patient suffering from pulmonary arterial hypertension comprises the steps of providing the patient with a dry powder inhaler, and providing the patient with at least one capsule for use in the dry powder inhaler, the capsule containing at least 25 micrograms of treprostinil.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Patent Application No. 62 / 332,013, filed May 5, 2016, U.S. Provisional Patent Application No. 62 / 404,960, filed October 6, 2016, U.S. Provisional Patent Application No. 62 / 440,078, filed December 29, 2016, and U.S. Provisional Patent Application No. 62 / 472,204, filed March 16, 2017, all of which are incorporated herein by reference in their entireties.

[0002] Technical Field

[0002] The present invention provides an improved treatment for pulmonary hypertension, a condition that worsens the lives of thousands of patients toward premature death. The present invention provides, for the first time, a stable, user-friendly, uniform dry powder inhaled treprostinil formulation, its method of manufacture, and its use in humans. [Background technology]

[0003] Pulmonary arterial hypertension (PAH) is a complex, multifactorial, progressive, and life-threatening disease characterized by proliferative and obstructive changes in the pulmonary vasculature, involving many biochemical pathways and cell types. The disease is characterized by elevated pulmonary arterial pressure caused by pulmonary vascular constriction and ultimately right ventricular failure. The disease has a poor prognosis associated with significant morbidity and mortality, with a historical survival rate of less than 5 years. PAH is a subgroup of pulmonary hypertension (PH) that increases pulmonary blood pressure. Endothelial dysfunction occurs early and is thought to result in cellular proliferation and structural changes in the pulmonary vasculature, leading to elevated pulmonary arterial pressure (PAP) and consequent right ventricular dilatation and dysfunction. Furthermore, endothelial dysfunction leads to chronic impairment of the production of vasoactive mediators such as nitric oxide (NO) and prostacyclin, as well as long-term overexpression of vasoconstrictors, such as endothelin-1.

[0004]

[0004] PAH affects approximately 15 out of every 1 million individuals. There are approximately 1,000 new cases of PAH diagnosed in the United States each year. The average age at diagnosis is 50 to 65 years, but the disorder can develop very early in childhood or even in infancy. Gender-based prevalence estimates for PAH vary, but estimates of the overall prevalence of pulmonary hypertension (PH) in women are approximately twice the prevalence in men.

[0005] PAH is part of a larger classification of pulmonary hypertension, which is divided into five groups (designated as WHO Groups 1 through 5) based on World Health Organization (WHO) criteria. PAH is used exclusively to describe WHO Group 1. Pulmonary hypertension is used to describe the remaining four groups (WHO Groups 2 through 5) and also to refer collectively to all five groups. · WHO Group 1-PAH: Pulmonary arterial hypertension. · WHO Group 2-PH: Pulmonary hypertension secondary to left heart disease. · WHO Group 3-PH: Pulmonary hypertension secondary to lung disease or hypoxemia. · WHO Group 4-PH: Chronic thromboembolic pulmonary hypertension. · WHO Group 5-PH: Pulmonary hypertension of unknown mechanism.

[0006]

[0006] PAH initially manifests as exertional dyspnea, lethargy, and fatigue, and is often confused with other pathologies. As PAH progresses and right ventricular failure develops, exertional chest pain (angina pectoris), exertional loss of consciousness, and peripheral edema may occur. Diagnosis based on hemodynamic parameters Once confirmed, treatment is recommended to reduce pulmonary pressure and treat the symptoms of PAH. Although there is no cure for PAH, treatment of PAH targets improvements in hemodynamic measures, New York Heart Association (NYHA) functional class, 6-minute walk distance (6MWD), quality of life, and, in some studies, survival.

[0007]

[0007] The severity of PAH can be classified according to the NYHA heart failure guidelines as follows: NYHA Class I: Patients with no activity limitations; patients do not suffer any symptoms from normal activities. NYHA Class II: Patients with minimal mild activity limitation; patients are comfortable at rest or with mild exertion. NYHA Class III: Patients with significant activity limitations; patients are only comfortable at rest. NYHA Class IV: Patients should be kept in bed or a chair and at complete rest; any physical activity causes discomfort and symptoms occur at rest.

[0008]

[0008] Although the exact underlying cause of PAH is unknown, mutations in the bone morphogenetic protein receptor type II (BMPR2) gene account for approximately 75% of familial PAH cases and up to 25% of apparently sporadic PAH cases. These mutations may promote cell division or prevent cell death, leading to overgrowth of cells in small pulmonary arteries. This overgrowth increases resistance to blood flow and causes hypertension. Additional genetic abnormalities may also contribute to PAH.

[0009] Currently Available Treatments

[0010] There are five classes of medications approved to treat PAH, including endothelin receptor antagonists (ERAs), phosphodiesterase type 5 (PDE5) inhibitors, soluble guanylate cyclase stimulators, prostacyclin receptor agonists, and prostacyclin analogs. Approved PAH therapies and their routes of administration include the following: ERA: bosentan (oral) and ambrisentan (oral) PDE5: Sildenafil (oral, intravenous (IV)) and tadalafil (oral) Soluble guanylate cyclase (sGC) stimulator: Riociguat (oral) Prostacyclin receptor agonist: Selexipag (oral) · Prostacyclin analogues: epoprostenol (IV), iloprost (inhaled), and treprostinil (oral), (subcutaneous and IV), and (inhaled).

[0010]

[0011] Treprostinil is a chemically stable tricyclic benzidine prostanoid with vasodilatory properties that can reduce pulmonary vasoconstriction with minimal effect on systemic blood pressure. Treprostinil is approved for the treatment of PAH under the trade names REMODULIN® (United Therapeutics Corporation; subcutaneous or IV infusion) and TYVASO® (United Therapeutics Corporation; inhaled via an ultrasonic, pulsed spray delivery device). While both have proven effective in PAH, one advantage of the inhaled administration route of TYVASO is that it delivers the drug very close to the desired site of action (the pulmonary artery within the lungs). Summary of the Invention [Problem to be solved by the invention]

[0011]

[0012] Despite current treatment options for patients with PAH, each option has drawbacks, most notably with regard to the inhalation route of administration, Tyvaso uses a large, cumbersome nebulizer device that requires power, water, and user manipulation for cleaning and operation. Furthermore, nebulized devices by their nature are not as convenient for patients as having small, concealable dry powder inhalation devices such as those used to treat asthma and many other chronic and acute conditions. Furthermore, nebulized treprostinil has shown clinical limitations in administering treprostinil, which may limit the applicability of the inhalation route of administration to a smaller subsector of PAH patients than the subsector of PAH patients necessarily treatable by inhalation from the dry powder inhaled treprostinil products of the present invention. [Means for solving the problem]

[0012]

[0013] The present inventors have developed and commercialized an inhalation dry powder formulation of treprostinil manufactured using Liquidia's PRINT® Technology (Particle Replication in Nonwetting Templates), LIQUIDIA TECHNOLOGIES, INC. This PRINT particle formulation (also referred to as LIQ861) for dry powder delivery of treprostinil is under clinical evaluation. Applicant has ...® technology is under clinical evaluation. The present invention contemplates the use of the same dosage and regimen (four times daily) for the same indication (i.e., treatment of pulmonary arterial hypertension (WHO Group 1) in patients with NYHA Class III symptoms to improve exercise capacity) as defined in the International Journal of Pulmonary Hypertension and Therapeutics (IJRA). In particular, the present invention provides for dosage levels that exceed the maximum tolerated dosage delivered via a nebulizer. In some cases, the present invention may also treat other indications under pulmonary hypertension conditions.

[0013]

[0014] In some embodiments, a dry powder inhalation therapy for pulmonary arterial hypertension according to the present invention comprises a dose of dry particles containing greater than 25 micrograms of treprostinil encapsulated in an encapsulated formulation. In some embodiments, the dose of dry particles comprises between about 25 micrograms and about 400 micrograms of treprostinil. In some embodiments, the dose of dry particles comprises between about 50 micrograms and about 350 micrograms of treprostinil. In some embodiments, the dose of dry particles comprises between about 75 micrograms and about 300 micrograms of treprostinil. In some embodiments, the dose of dry particles comprises between about 100 micrograms and about 300 micrograms of treprostinil. In some embodiments, the dose of dry particles comprises 100 micrograms or more of treprostinil. In some embodiments, the dose of dry particles comprises 150 micrograms or more of treprostinil. In some embodiments, the dose of dry particles comprises 200 micrograms or more of treprostinil. In some embodiments, the dose of dry particles comprises 250 micrograms or more of treprostinil. In some embodiments, the dose of dry particles comprises 300 micrograms or more of treprostinil. In some embodiments, the dose of dry particles comprises 5 mg or more of dry particles. In some embodiments, the dose of dry particles comprises 10 mg or more of dry particles. In still other embodiments, the dose of dry particles comprises 15 mg or more of dry particles. In a further embodiment, the dry powder treatment for pulmonary arterial hypertension comprises a single capsule encapsulating 5 mg or more of dry particles, each 5 mg of dry particles containing 25 micrograms of treprostinil.

[0014]

[0015] In some embodiments, a method of treating a patient suffering from pulmonary arterial hypertension includes providing the patient with a dry powder inhaler; providing the patient with at least one capsule for use in the dry powder inhaler, the capsule containing at least 25 micrograms of treprostinil; and instructing the patient to inhale the treprostinil using the dry powder inhaler. In some such embodiments, the capsule contains at least 50 micrograms of treprostinil. In some embodiments, the capsule contains at least 100 micrograms of treprostinil. In some embodiments, the capsule contains at least 150 micrograms of treprostinil. In some embodiments, the capsule contains 200 micrograms or more of treprostinil. In some embodiments, the capsule contains 250 micrograms or more of treprostinil. In some embodiments, the capsule contains 300 micrograms or more of treprostinil. In some embodiments, the capsule contains about 25 micrograms to about 400 micrograms of treprostinil. In some embodiments, the capsule contains about 50 micrograms to about 350 micrograms of treprostinil. In some embodiments, the capsule contains about 75 micrograms to about 300 micrograms of treprostinil. In some embodiments, the capsule contains about 100 micrograms to about 300 micrograms of treprostinil. In further embodiments, patients may be prescribed two capsules per dosing cycle per day, with PAH typically requiring four doses per day. In some embodiments, a patient may be prescribed three capsules per day. In some embodiments, a patient may be prescribed four capsules per day. In some embodiments, a method for treating a patient suffering from pulmonary arterial hypertension includes administering a dry powder dose of treprostinil to a patient suffering from pulmonary arterial hypertension, wherein the dose of treprostinil is greater than 85 micrograms (e.g., about 100 micrograms to about 350 micrograms). In some embodiments, the patient may be administered one, two, three, four, or more doses per day. A further method for treating a patient suffering from pulmonary arterial hypertension includes delivering greater than 12.5 micrograms of treprostinil per breath to a patient via a dry powder. In another embodiment, a method for treating a patient suffering from pulmonary arterial hypertension includes delivering greater than 25 micrograms of treprostinil per breath to a patient via a dry powder.In another embodiment, a method for treating a patient suffering from pulmonary arterial hypertension comprises delivering about 12.5 to about 50 micrograms of treprostinil per breath to the patient as a dry powder. In yet another embodiment, a method for treating a patient suffering from pulmonary arterial hypertension comprises delivering about 25 to about 50 micrograms of treprostinil per breath to the patient as a dry powder. In a further embodiment, a method for treating a patient suffering from pulmonary arterial hypertension comprises delivering more than 50 micrograms of treprostinil per breath to the patient as a dry powder. In a further embodiment, a method for treating a patient suffering from pulmonary arterial hypertension comprises delivering 100 micrograms or more of treprostinil per breath to the patient as a dry powder. In a further embodiment, a method for treating a patient suffering from pulmonary arterial hypertension comprises delivering 150 micrograms or more of treprostinil per breath to the patient as a dry powder. In a further embodiment, a method of treating a patient suffering from pulmonary arterial hypertension comprises delivering 200 micrograms or more of treprostinil per breath to the patient in a dry powder.

[0015]

[0016] Further embodiments of a dry powder inhalation composition for treating pulmonary arterial hypertension include a plurality of dry powder particles comprising treprostinil, a non-reducing sugar, a humectant, a hydrophobicity modifier, a pH adjuster, and a buffer. In some such embodiments, the bulking agent comprises trehalose dihydrate. In some embodiments, the humectant comprises polysorbate 80. In some embodiments, the hydrophobicity modifier comprises L-leucine. In some embodiments, the pH adjuster comprises sodium citrate dihydrate. In some embodiments, the buffer comprises sodium chloride. In certain embodiments, the composition comprises less than about 4 percent water by weight. In some embodiments, the composition comprises less than about 2 percent water by weight. In some embodiments, the composition comprises less than about 1 percent water by weight.

[0016]

[0017] In still further embodiments, the dry powder particles comprise particles having a three-dimensional shape comprising a width and length of 1 to 2 micrometers, and a depth of 0.3 to 0.8 micrometers. In some embodiments, the dry powder particles comprise a dry solution comprising trehalose dihydrate, L-leucine, treprostinil sodium, polysorbate 80, sodium citrate dihydrate, sodium chloride, and water. In some embodiments, the dry powder particles comprise, in solids percent, about 0.581 percent treprostinil sodium, about 92.32 percent trehalose, about 2.19 percent polysorbate 80, about 4.39 percent L-leucine, about 0.26 percent sodium citrate, and about 0.25 percent sodium chloride.

[0017]

[0018] In some embodiments, a method for producing particles for dry powder delivery to the lungs of a patient in need thereof includes forming particles by placing a composition comprising about 12.30 weight percent trehalose dihydrate, about 0.53 weight percent L-leucine, about 0.07 weight percent treprostinil sodium, about 0.26 weight percent polysorbate 80, about 0.04 weight percent sodium citrate dihydrate, about 0.03 weight percent sodium chloride, and about 86.78 weight percent water in a mold. In some embodiments, the method for producing particles further includes drying the composition such that the particles contain less than 4 weight percent water.

[0018]

[0019] The foregoing summary, as well as the following detailed description of the invention, will be better understood when read in conjunction with the accompanying drawings. For the purpose of illustrating the invention, there are shown in the drawings embodiments which are presently preferred. It should be understood, however, that the invention may be embodied in different forms and, therefore, should not be construed as limited to the illustrated embodiments set forth herein. [Brief explanation of the drawings]

[0019] [Figure 1]

[0020] FIG. 1 illustrates a three-dimensional representation of a pollen particle according to one embodiment of the present invention. [Figure 2]

[0021] Figure 1 shows an example of NGI distribution of active particles (PAH-1R-0943-010). For each of the three data sets shown for each collection cup, the start of the run is the left bar (A1), the middle bar (B1), and the end of the run is the right bar (C1). Data were acquired using a Monodose Model 8 device (95 L / min, 2 seconds). [Figure 3A]

[0022] 3A and 3B are tables containing data for Cohort 1 of the clinical trial. The table shown in FIG. 3A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 1. The table shown in FIG. 3B summarizes pre-study non-compartmental PK parameters for treprostinil. [Figure 3B]

[0022] Tables containing data for Cohort 1 of the clinical trial. The table shown in Figure 3A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 1. The table shown in Figure 3B summarizes pre-study non-compartmental PK parameters for treprostinil. [Figure 4A]

[0023] 4A and 4B are tables containing data for Cohort 2 of the clinical trial. The table shown in FIG. 4A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 2. The table shown in FIG. 4B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 2. [Figure 4B] Tables containing data for Cohort 2 of the clinical trial. The table shown in Figure 4A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 2. The table shown in Figure 4B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 2. [Figure 5A]

[0024] 5A and 5B are tables containing data for Cohort 3 of the clinical trial. The table shown in FIG. 5A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 3. The table shown in FIG. 5B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 3. [Figure 5B]

[0024] Tables containing data for Cohort 3 of the clinical trial. The table shown in Figure 5A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 3. The table shown in Figure 5B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 3. [Figure 6A]

[0025] 6A is a table containing data for Cohort 4 of the clinical trial. The table shown in FIG. 6A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 4. FIG. 6B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 4. [Figure 6B] Tables containing data for Cohort 4 of the clinical trial. The table shown in Figure 6A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 4. Figure 6B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 4. [Figure 7A]

[0026] 7A and 7B are tables containing data for Cohort 5 of the clinical trial. The table shown in FIG. 7A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 5. FIG. 7B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 5. [Figure 7B]Tables containing data for Cohort 5 of the clinical trial. The table shown in Figure 7A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 5. Figure 7B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 5. [Figure 8A]

[0027] 8A and 8B are tables containing data for Cohort 6 of the clinical trial. The table shown in FIG. 8A includes a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 6-R. FIG. 8B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 6-R. FIG. 8C summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 6-Original. [Figure 8B]

[0027] Tables containing data for Cohort 6 of the clinical trial. The table shown in Figure 8A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 6-R. Figure 8B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 6-R. Figure 8C summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 6-Original. [Figure 8C]

[0027] Tables containing data for Cohort 6 of the clinical trial. The table shown in Figure 8A contains a summary of treprostinil concentration-time data for individual subjects, along with descriptive statistics for the six eligible subjects in Cohort 6-R. Figure 8B summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 6-R. Figure 8C summarizes pre-study non-compartmental PK parameters for treprostinil in Cohort 6-Original. [Figure 8D]

[0028] Figures 8A and 8B contain data from clinical trials. Figure 8D shows the mean concentration-time data for each of the six cohorts on a linear scale. Figures 8E and 8F show plots of the relationship between dose and Cmax and AUCinf, respectively. Figure 8G shows plots of the relationship between dose and oral clearance CL / F. [Figure 8E]

[0028] Figures containing data from clinical trials. Figure 8D shows the mean concentration-time data for each of the six cohorts on a linear scale. Figures 8E and 8F show plots of the relationship between dose and Cmax and AUCinf, respectively. Figure 8G shows plots of the relationship between dose and oral clearance CL / F. [Figure 8F]

[0028] Figures containing data from clinical trials. Figure 8D shows the mean concentration-time data for each of the six cohorts on a linear scale. Figures 8E and 8F show plots of the relationship between dose and Cmax and AUCinf, respectively. Figure 8G shows plots of the relationship between dose and oral clearance CL / F. [Figure 8G]

[0028] Figures containing data from clinical trials. Figure 8D shows the mean concentration-time data for each of the six cohorts on a linear scale. Figures 8E and 8F show plots of the relationship between dose and Cmax and AUCinf, respectively. Figure 8G shows plots of the relationship between dose and oral clearance CL / F. [Figure 9]

[0029] 1 is an SEM image showing a pollen particle according to an embodiment of the present invention. [Figure 10]

[0030] FIG. 1 is a flow diagram illustrating a process for producing particles according to one embodiment of the present invention. [Figure 11]

[0031] 1 illustrates an example dry powder inhalation device that can be used to deliver particles to a patient in accordance with embodiments of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0020]

[0032] drug substance

[0033] The drug substance (DS) according to an embodiment of the present invention is treprostinil, which is a synthetic analogue of prostacyclin (PGI2). The IUPAC name for treprostinil is (2-[[(1R,2R,3aS,9aS)-2-hydroxy-1-[(3S)-3-hydroxyoctyl]-2,3,3a,4,9,9a-hexahydro-1H-cyclopenta[g]naphthalen-5-yl]oxy]acetic acid).

[0021]

[0034] Powdered inhalation medication

[0035] Certain aspects of the present invention provide inhalable powder formulations (formulation intermediates or DP intermediates) of treprostinil and excipients formed into particles, which in some embodiments are filled into capsules, such as hydroxypropylmethylcellulose (HPMC) capsules (size 3) (LIQ861). In some embodiments, the DP intermediates are treprostinil / excipient matrices formed by the methods herein into particles of precise size and shape. In one example, the DP intermediate particles have a shape generally corresponding to a rounded triangular shape with a volume, where the interior of the rounded triangular shape fits into an equilateral triangle of 1 micrometer in size (also referred to as a pollen shape). A three-dimensional representation of such a particle shape is shown in Figure 1. In another embodiment, the pollen shape may be a trilobal shape with an inscribed circle diameter of 1 micrometer and a predetermined thickness of 0.5 to 1 micrometer, more preferably 0.7 micrometer. Additionally, certain embodiments of the medicament include particles with 0.5% treprostinil used in initial clinical trials investigating dosage levels of 25mcg, 50mcg, 75mcg, 100mcg, 125mcg, and 150mcg treprostinil in LIQ 861. In further embodiments, medicaments according to the invention may provide dosage levels of 175mcg, 200mcg, 225mcg, 250mcg, 275mcg, 300mcg, 325mcg, or 350mcg treprostinil. In a further embodiment, the medicament according to the invention may provide a dosage level of 50mcg of treprostinil plus or minus 10mcg, 9mcg, 8mcg, 7mcg, 6mcg, 5mcg, 4mcg, 3mcg, 2mcg, or 1mcg of treprostinil loaded into a capsule for delivery to a patient as a dry powder. In a further embodiment, the medicament according to the invention may provide a dosage level of 75mcg of treprostinil plus or minus 10mcg, 9mcg, 8mcg, 7mcg, 6mcg, 5mcg, 4mcg, 3mcg, 2mcg, or 1mcg of treprostinil loaded into a capsule for delivery to a patient as a dry powder.In a further embodiment, the medicament according to the invention is a dose of 100mcg of treprostinil plus or minus 10mcg, 9mcg, 8mcg, 7mcg, 6mcg, 5mcg, 4mcg, 3mcg, 2mcg or 1mcg treprostinil loaded into a capsule for delivery to a patient as a dry powder. In a further embodiment, the medicament according to the present invention can be provided at a dosage level of 150 mcg of treprostinil plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% treprostinil packed into a capsule for delivery to a patient as a dry powder. In a further embodiment, the medicament according to the present invention can be provided at a dosage level of 200 mcg of treprostinil plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% treprostinil packed into a capsule for delivery to a patient as a dry powder. In a further embodiment, the medicament according to the present invention can be provided at a dosage level of 300 mcg of treprostinil plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% treprostinil packed into a capsule for delivery to a patient as a dry powder.

[0022]

[0036] According to the present invention, the dry powder particle formulation allows the particles to remain stable for extended periods of time under relatively low humidity conditions. In some embodiments, the present invention provides dry powder particles packaged under sealed conditions that maintain stability for at least three months at 75 percent relative humidity and 40 degrees Celsius. Thus, the particles can be used to provide patients with a dry powder inhalation drug form of treprostinil that was previously unavailable. In some embodiments, the present invention provides users with reduced drug exposure by eliminating the need for patients to reconstitute their drug for use in a nebulizer device. It also allows patients to receive a dose equivalent to a greater than 50 percent reduction in breath therapy with the device, and in some embodiments, a greater than 65 percent reduction in breath therapy.

[0023]

[0037] The present invention also provides, in some embodiments, dry formulations of treprostinil that, when delivered to a patient via the inhalation route, become soluble and pharmaceutically available in less than 10 seconds. In some embodiments, the dry formulation composition becomes soluble and pharmaceutically available in less than 5 seconds. In some embodiments, the dry formulation composition becomes soluble and pharmaceutically available in less than 2 seconds. In some embodiments, the dry formulation composition becomes soluble and pharmaceutically available in about 1 second. In some embodiments, the dry formulation composition becomes soluble and pharmaceutically available in less than 1 second. In some embodiments, the dry formulation composition becomes soluble and pharmaceutically available in less than about 0.5 seconds. Furthermore, the excipients in the dry particle formulations of the present invention maintain pH and salt gradients during the process, allowing the active agent to remain in a state that is soluble in the user's pulmonary conditions.

[0024]

[0038] A detailed description of the LIQ861 formulation, particle composition, particle geometry, packaging, device, delivery, stability, dosage, and instructions for use follows.

[0039] In some embodiments, formulations according to the invention comprise a drug substance (e.g., treprostinil, treprostinil sodium) together with one or more excipients. In some embodiments, the one or more excipients may comprise a bulking agent, a wetting agent, a hydrophobicity adjusting agent, a pH adjusting agent, a buffering component, or a combination thereof. Examples of such formulations according to certain embodiments are provided in the table below.

[0025] [Table 1]

[0026] [Table 2]

[0027]

[0040] Inhalation devices

[0041] According to one embodiment of administering the drug particles of the invention, LIQ861 is administered using an RS00 Model 8 dry powder inhaler device (Plastiape SpA). The invention provides for multi-day administration of LIQ861, according to some embodiments.

[0028]

[0042] Indications

[0043] The present invention, according to one embodiment, is useful for the treatment of pulmonary arterial hypertension (WHO Group 1) in patients with NYHA Class III symptoms to improve exercise capacity.

[0029]

[0044] Chemistry, Manufacturing and Controls (CMC)

[0045] Drug Substances (DS)

[0046] The drug substance according to an embodiment of the present invention is treprostinil, and the salt form used in LIQ861 is treprostinil sodium. Detailed information about treprostinil sodium, including its physical and chemical properties, characteristics, manufacturing and controls, container closure system, and stability attributes, can be found in the Drug Master File (DMF) submitted to the FDA for treprostinil. General information regarding the DS is provided herein.

[0030]

[0047] naming

[0048] The International Nonproprietary Name (INN) for LIQ861 is treprostinil sodium. The chemical name is 2-((1R,2R,3aS,9aS)-2-hydroxy-1-((S)-3-hydroxyoctyl)-2,3,3a,4,9,9a-hexahydro-1H-cyclopenta[b]naphthalen-5-yloxy)acetic acid, sodium salt. The Chemical Abstracts Service registry number is [289480-64-4].

[0031]

[0049] structure

[0050] The structure of treprostinil sodium is shown herein below. The molecular formula is C 23 H 33NaO5 and has a molecular weight of 412.49 daltons.

[0032]

[0051] Chemical structure of treprostinil sodium

[0033] [ka]

[0034]

[0052] General characteristics

[0053] Treprostinil sodium appears as a white or pale yellow powder. Treprostinil sodium is very soluble in water and ethanol, sparingly soluble in acetone, and practically insoluble in acetonitrile, n-hexane, and ethyl acetate. The specific optical rotation, calculated on an anhydrous and solvent-free basis, is [α] D 20 = +38.0° to +44.0°. It is hygroscopic. The pKa of treprostinil is 4.5 using aqueous titration with 20% ethanol as a cosolvent. Distribution coefficients of treprostinil in various buffer solutions at various pH levels show distribution into the octanol layer at all pH levels.

[0035]

[0054] Inhaled particle formulation - LIQ861

[0055] Description and composition of the formulation particles

[0056] Inhaled drug particle products, in some embodiments, comprise or consist of dry powder formulations (drug intermediates; DP intermediates; or drug particles) of treprostinil and excipients, which may be filled, for example, into HPMC capsules (size 3). The DP intermediates, in some embodiments, are treprostinil / excipient matrices in which particles of precise size (e.g., 1 μm) and shape (e.g., "pollen-shaped") are generated using Liquidia's PRINT technology. "Pollen-shaped" particles can also be described as trilobe-shaped, with an inscribed circle diameter of 1 μm and a thickness of 0.7 μm. A three-dimensional representation of such particle shapes is shown in Figure 1. LIQ861 contained formulation capsule strengths of 25 mcg, 50 mcg, and 75 mcg of treprostinil used in initial clinical trials investigating planned dosage levels of treprostinil of 25 mcg, 50 mcg, 75 mcg, 100 mcg, 125 mcg, and 150 mcg. The 100mcg, 125mcg, and 150mcg doses may also consist of a combination of lower dose capsules. In a further embodiment, the medicament according to the invention may be provided in capsules at dosage levels of 175mcg, 200mcg, 225mcg, 250mcg, 275mcg, 300mcg, 325mcg, or 350mcg treprostinil. In a further embodiment, the medicament according to the invention may be provided in capsules at dosage levels of 50mcg treprostinil plus or minus 10mcg, 9mcg, 8mcg, 7mcg, 6mcg, 5mcg, 4mcg, 3mcg, 2mcg, or 1mcg treprostinil for delivery to a patient in a dry powder. In a further embodiment, the medicament according to the invention may be provided in capsules at dosage levels of 75mcg treprostinil plus or minus 10mcg

[0039] In a further embodiment, the medicament according to the present invention can be provided in capsules at dosage levels of 100mcg treprostinil plus or minus 10mcg, 9mcg, 8mcg, 7mcg, 6mcg, 5mcg, 4mcg, 3mcg, 2mcg, or 1mcg treprostinil for delivery to a patient as a dry powder. In a further embodiment, the medicament according to the present invention can be provided in capsules at dosage levels of 150mcg treprostinil plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% treprostinil for delivery to a patient as a dry powder. In a further embodiment, the medicament according to the present invention can be provided in capsules at a dosage level of 200 mcg treprostinil plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% mcg treprostinil for delivery to a patient as a dry powder. In a further embodiment, the medicament according to the present invention can be provided in capsules at a dosage level of 300 mcg treprostinil plus or minus 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% treprostinil for delivery to a patient as a dry powder. A summary of the LIQ861 formulation, including the powder composition, particle geometry, and dosage unit description according to certain exemplary embodiments, is provided below.

[0036] [Table 3]

[0037] [Table 4]

[0038]

[0057] According to some embodiments of the present invention, drug particles are provided comprising a composition having a target dose of treprostinil delivered to a patient of 15-90 μg (currently labeled TYVASO® as 18-54 μg). In some embodiments of the present invention, the dose of treprostinil provided to a patient may be, for example, 100 micrograms, 125 micrograms, or 150 micrograms. In some embodiments of the present invention, the dose of treprostinil provided to a patient may be, for example, about 100 micrograms, about 125 micrograms, or about 150 micrograms. In some embodiments, each dose contains 200 micrograms or more of treprostinil. In some embodiments, each dose contains 225 micrograms or more of treprostinil. In some embodiments, each dose contains 250 micrograms or more of treprostinil. In some embodiments, each dose contains 275 micrograms or more of treprostinil. In some embodiments, each dose contains 300 micrograms or more of treprostinil. In some embodiments, each dose contains about 10 micrograms to about 15 micrograms, 15 micrograms to about 20 micrograms, 20 micrograms to about 25 micrograms, 25 micrograms to about 30 micrograms, about 30 micrograms to about 35 micrograms, about 35 micrograms to about 40 micrograms, about 40 micrograms to about 45 micrograms, about 45 micrograms to about 50 micrograms, about 50 micrograms to about 55 micrograms, about 55 micrograms to about 60 micrograms, about 60 micrograms to about 65 micrograms, about 65 micrograms to about 70 micrograms, about 70 micrograms to about 75 micrograms, about 75 micrograms to about 80 micrograms, about 80 micrograms to about 85 micrograms, about 85 micrograms to about 90 micrograms, about 90 micrograms to about 95 micrograms, about 95 micrograms to about 100 micrograms, or about 100 micrograms to about 105 micrograms of treprostinil.In some embodiments, each dose is from about 100 micrograms to about 110 micrograms, 110 micrograms to about 120 micrograms, 120 micrograms to about 130 micrograms, 130 micrograms to about 140 micrograms, about 140 micrograms to about 150 micrograms, about 150 micrograms to about 160 micrograms, about 160 micrograms to about 170 micrograms, about 170 micrograms to about 180 micrograms, about 180 micrograms to about 190 micrograms, about 190 micrograms to about 200 micrograms, about 200 micrograms to about 210 micrograms, about 210 micrograms to about 220 micrograms, or about 220 micrograms to about 150 micrograms. In some embodiments, each dose contains about 230 micrograms, about 230 micrograms to about 240 micrograms, about 240 micrograms to about 250 micrograms, about 250 micrograms to about 260 micrograms, about 260 micrograms to about 270 micrograms, about 270 micrograms to about 280 micrograms, about 280 micrograms to about 290 micrograms, about 290 micrograms to about 300 micrograms, about 300 micrograms to about 310 micrograms, about 310 micrograms to about 320 micrograms, about 320 micrograms to about 330 micrograms, about 330 micrograms to about 340 micrograms, or about 340 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 25 micrograms to about 400 micrograms of treprostinil. In some embodiments, each dose contains about 25 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 25 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 50 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 150 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 175 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 200 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 225 micrograms to about 300 micrograms of treprostinil.In some embodiments, each dose contains about 250 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 275 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 50 micrograms to about 75 micrograms of treprostinil. In some embodiments, each dose contains about 50 micrograms to about 100 micrograms of treprostinil. In some embodiments, each dose contains about 50 micrograms to about 150 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 100 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 125 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 150 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 175 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 200 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 225 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 250 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 275 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 75 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 125 micrograms of treprostinil.In some embodiments, each dose contains about 100 micrograms to about 150 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 175 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 200 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 225 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 250 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 275 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 100 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 150 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 175 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 200 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 225 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 250 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 275 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 125 micrograms to about 350 micrograms of treprostinil.In some embodiments, each dose contains about 150 micrograms to about 175 micrograms of treprostinil. In some embodiments, each dose contains about 150 micrograms to about 200 micrograms of treprostinil. In some embodiments, each dose contains about 150 micrograms to about 225 micrograms of treprostinil. In some embodiments, each dose contains about 150 micrograms to about 250 micrograms of treprostinil. In some embodiments, each dose contains about 150 micrograms to about 275 micrograms of treprostinil. In some embodiments, each dose contains about 150 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 150 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 150 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 175 micrograms to about 200 micrograms of treprostinil. In some embodiments, each dose contains about 175 micrograms to about 225 micrograms of treprostinil. In some embodiments, each dose contains about 175 micrograms to about 250 micrograms of treprostinil. In some embodiments, each dose contains about 175 micrograms to about 275 micrograms of treprostinil. In some embodiments, each dose contains about 175 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 175 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 175 micrograms to about 350 micrograms of treprostinil. In some embodiments. In some embodiments, each dose contains about 200 micrograms to about 225 micrograms of treprostinil. In some embodiments, each dose contains about 200 micrograms to about 250 micrograms of treprostinil. In some embodiments, each dose contains about 200 micrograms to about 275 micrograms of treprostinil. In some embodiments, each dose contains about 200 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 200 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 200 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 225 micrograms to about 250 micrograms of treprostinil. In some embodiments, each dose contains about 225 micrograms to about 275 micrograms of treprostinil. In some embodiments, each dose contains about 225 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 225 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 225 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 250 micrograms to about 275 micrograms of treprostinil. In some embodiments, each dose contains about 250 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 250 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 250 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 275 micrograms to about 300 micrograms of treprostinil. In some embodiments, each dose contains about 275 micrograms to about 325 micrograms of treprostinil. In some embodiments, each dose contains about 275 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 300 micrograms to about 325 micrograms of treprostinil.In some embodiments, each dose contains about 300 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 325 micrograms to about 350 micrograms of treprostinil. In some embodiments, each dose contains about 350 micrograms to about 375 micrograms of treprostinil. In some embodiments, each dose contains about 375 micrograms to about 400 micrograms of treprostinil. In some embodiments, patients may be provided with one, two, three, four, or more doses per day. In some embodiments, patients may be provided with up to one, two, three, or four doses per day. Each dose may be contained in a single capsule, e.g., an HPMC capsule (size 3), according to some embodiments. In other embodiments, a dose may consist of a combination of lower dose capsules. In some embodiments, patients can be provided with four doses per day to match their current treatment cycle (nebulized treprostinil), but the drug dosage per treatment cycle under the dry powder of the present invention surprisingly provides significantly higher dosage levels that can be safely administered, e.g., 100 mcg or less of treprostinil per dose, 125 mcg or less of treprostinil per dose, and 150 mcg or less of treprostinil per dose, as demonstrated in the first phase clinical trials of LIQ861. In alternative embodiments, patients can be provided with four doses per day to match their current treatment cycle (nebulized treprostinil), but the drug dosage per treatment cycle under the dry powder of the present invention surprisingly provides significantly higher dosage levels that can be achieved, e.g., 200 mcg or less of treprostinil per dose, and 300 mcg or less of treprostinil per dose, as demonstrated in preclinical toxicity studies using LIQ861.

[0039]

[0058] Treprostinil itself is poorly soluble in unbuffered water and low pH buffers, but solubility improves with increasing pH as the carboxylic acid becomes deprotonated. The sodium salt was chosen for use in this product because it promotes solubility in aqueous media and eases processing.

[0040]

[0059] excipients

[0060] According to some embodiments of the present invention, the DP intermediate (anhydrous) includes particles containing, for example, the following excipients: trehalose, polysorbate 80, L-leucine, sodium citrate, and sodium chloride. In some embodiments, the ratio of treprostinil sodium to excipients is 0.581:92.32:2.19:4.39:0.26:0.25 (wt:wt solids) for treprostinil sodium:trehalose:polysorbate 80:leucine:sodium citrate:sodium chloride. A summary of the function, amount, and general status of these excipients is provided herein.

[0041]

[0061] The excipients were selected based on the following functional requirements for the formulation: Trehalose dihydrate: Trehalose provides the bulk of the particles and was selected because it is a non-reducing sugar with a high glass transition temperature. Trehalose is an example of a non-reducing sugar (unlike lactose, which is a reducing sugar) that can be used in the present invention. Trehalose is more chemically compatible with compounds containing primary amines, such as leucine. Ultra-Pure Polysorbate 80 (Ultra-Pure Tween 80): Polysorbate 80 is added as a processing aid / wetting agent to facilitate particle manufacturing. In some embodiments, Polysorbate 80 is a particle processing aid, reducing dewetting so that films are formed during particle manufacturing, resulting in uniform particle morphology. L-Leucine: Leucine is added as a hydrophobicity and surface modifier to reduce particle hygroscopicity and improve aerosol efficiency. L-Leucine is an example of a formulation additive used to reduce hygroscopicity and improve the stability of the final drug powder. Sodium Chloride and Sodium Citrate: Sodium citrate and sodium chloride are used to buffer the stock solutions used in the PRINT technology manufacturing process and to adjust the acidity of the particles. Sodium chloride and sodium citrate are examples of buffering agents that help maintain pH and adjust the ionization / acidity of the formulation. In some embodiments of the present invention, the pH is maintained between about pH 6.0 and 7.2.

[0042]

[0062] In addition to the active pharmaceutical ingredient, the drug particles of the present invention comprise a filler, a wetting agent, a hydrophobicity modifier, a pH adjuster, and a buffering agent. In some embodiments, the drug particles of the present invention comprise a filler, a hydrophobicity modifier, and a pH adjuster along with the active ingredient.

[0043]

[0063] According to another embodiment of the present invention, LIQ861 comprises five excipients as follows: treprostinil sodium:trehalose dihydrate:leucine:polysorbate 80:sodium citrate dihydrate:sodium chloride in the ratios of 0.53:92.97:4:2:0.27:0.23. In one example of a 100 μg / day treprostinil dosage level of drug particles of the present invention, a patient receives the following daily excipient dosages: 18.6 mg of trehalose dihydrate. Assuming a patient weighing 60 kg and a lung mass of 1000 g, this corresponds to 310 μg / kg and 18.6 μg / g of lung. 0.4 mg of polysorbate 80. Assuming a patient weight of 60 kg and a lung mass of 1000 g, this corresponds to 6.7 μg / kg and 0.4 μg / g of lung. 0.8 mg of leucine. Assuming a patient weighing 60 kg and a lung mass of 1000 g, this corresponds to 13.3 μg / kg and 0.8 μg / g of lung. 0.05 mg sodium citrate and 0.05 mg sodium chloride. Assuming the patient weighs 60 kg and the lung mass is 1000 g, this is This corresponds to 0.83 μg / kg for the pulmonary route and 0.05 μg / g for each compound.

[0044]

[0064] Formulation Development

[0065] According to an embodiment of the present invention, LIQ861 has been developed as a novel formulation of treprostinil for treating PAH. Treprostinil is currently approved for use in the treatment of PAH by subcutaneous, intravenous, oral, and inhalation routes. TYVASO is currently the only commercially available inhalation formulation of treprostinil, formulated as a liquid solution administered using a nebulizer. Nebulized treprostinil is administered at a maintenance dose of 6 mcg of drug per breath over 9 breaths to deliver a dose of 54 mcg per administration session. Nebulized treprostinil also has a maximum tolerated dose of 84 mcg over a 14-breath administration session.

[0045]

[0066] LIQ861 is suitable for inhalation administration using a dry powder inhaler device. The physicochemical and performance characteristics, manufacturing process and packaging, and stability characteristics of the DP have been tested to identify suitable formulations for deployment in human trials.

[0046]

[0067] Physiochemical and biological properties

[0068] "Pollen-shaped" LIQ861 particles according to certain embodiments have an aerodynamic size (1≦MMAD≦5 μm) that allows efficient delivery to pulmonary arterioles with a high FPF to limit oropharyngeal deposition. A scanning electron microscope (SEM) image of the "pollen-shaped" features is shown in FIG. 9. The formulation of the exemplary particles shown in FIG. 9 is Treprostinil:Trehalose:Leucine:Polysorbate 80:Sodium Citrate:Sodium Chloride (Batch LKI-1R-983-27). Example aerosol data for the active particles are also shown in the table below.

[0047]

[0069] During development of the LIQ861 formulation, Applicants also tested other potential particle shapes and sizes (e.g., 1.5 μm donut-shaped, 3.0 μm donut-shaped). Based on these tests, Applicants observed that the "pollen-shaped" profile provided increased FPF, lower MMAD, acceptable ED, and uniform dosage characteristics when compared to other profiles with or without treprostinil.

[0048] [Table 5]

[0049]

[0070] manufacturing

[0071] The preparation of LIQ861 particles according to some embodiments of the present invention is described below. A process flow diagram for particles (also referred to as DP intermediates) according to some embodiments is shown in FIG.

[0050]

[0072] In certain embodiments, the particles of the present disclosure are manufactured using PRINT® technology (Liquidia Technologies, Inc., Morrisville, NC) particle manufacturing. In particular, the particles are manufactured by molding a material to form the particles in a mold cavity.

[0051]

[0073] In some embodiments, the mold may be a polymer-based mold, and the mold cavity can be formed into any desired shape and size. Uniquely, because the particles are formed in the mold cavity, the particles are highly uniform in shape, size, and composition. Due to the consistency between the physical and compositional makeup of the particles of the present composition, the composition of the present disclosure provides a highly uniform release rate and dosage range. Methods and materials that can be used to manufacture particles according to embodiments of the present disclosure are further described and disclosed in issued patents and co-pending patent applications, each of which is incorporated herein by reference in its entirety: U.S. Patent Nos. 8,518,316; 8,444,907; 8,420,124; 8,268,446; 8,263,129; 8,158,728; 8,128,393; 7,976,759; U.S. Patent Application Publication No. 2013 Nos. 2013-0249138, 2013-0241107, 2013-0228950, 2013-0202729, 2013-0011618, 2013-0256354, 2012-0189728, 2010-0003291, 2009-0165320, 2008-0131692; and pending U.S. application Ser. Nos. 13 / 852,683, filed March 28, 2013, and 13 / 950,447, filed July 25, 2013.

[0052]

[0074] Particle production

[0075] An aqueous stock solution is prepared at the desired total solids concentration. All other excipients are combined with the treprostinil and then filtered before producing the particles.

[0053]

[0076] The stock solution is applied in a thin layer to a continuous polyethylene terephthalate (PET) substrate backing layer. Forced air heat is used to remove water, resulting in a dry film of treprostinil and excipients. The dried film is then contacted at elevated temperature with a mold film having cavities of the desired shape and size that the formulation particles will mimic. The drug / excipient mixture flows into the mold cavities and conforms to the shape defined by the cavities. The result is a uniform array of particles attached to the PET backing layer. The particles are then cooled to room temperature so that the roll can be wound for later collection.

[0054]

[0077] In one example of a drug particle of the present invention, the following stock solution is used: Ingredients of the stock solution used in the manufacture of treprostinil particles according to one embodiment:

[0055] [Table 6]

[0056]

[0078] Dry recovery and drying

[0079] The particles are then dried and collected, and the shaped particles are removed from the PET backing layer, thereby producing a bulk powder. The molded object is first separated from the PET backing layer, exposing the particle array attached to the PET backing layer. The particle array is then passed across a blade, in some embodiments a plastic blade, to remove the particles from the backing layer. The particles can then be collected into a bulk powder for further processing.

[0057]

[0080] Humidity is controlled to less than 15% RH during collection in some embodiments due to the hygroscopic nature of the powder. Temperature is maintained at ambient temperature, typically 15-25°C.

[0081] Dry and bulk packaging

[0082] The drug particles, according to some embodiments, are subjected to nitrogen or Dry in dry air in a benchtop freeze dryer at room temperature for at least 2 days.

[0058]

[0083] In some embodiments, the particles of the present invention are dried to a moisture content of less than about 10%. In some embodiments, the particles of the present invention are dried to a moisture content of less than about 5%. In further embodiments, the particles of the present invention are dried to a moisture content of less than about 4%. In still further embodiments, the particles of the present invention are dried to a moisture content of less than about 2%. In a preferred embodiment, the product is dried to a moisture content of less than about 1% by Karl Fischer titration.

[0059]

[0084] Batch-to-batch uniformity of drug particles

[0085] In some embodiments, batch-to-batch particle uniformity provides the present invention with unexpected advantages over the prior art. In certain embodiments, any given within-batch uniformity is unexpected and highly advantageous over the prior art. The present invention includes highly consistent batch uniformity, as shown in the following data. See also the table below and Figure 2.

[0060] [Table 7]

[0061] In the example shown, the fine particle fraction was maintained within plus / minus 1% in a single batch run.

[0086] Capsule Filling and Packaging

[0087] In some embodiments, HPMC capsules are filled with DP intermediates in a humidity-controlled ISO8 environment using an XCELODOSE® (Capsugel) machine. The filled HPMC capsules are packaged in a low-humidity environment. Ten capsules are placed in a DESICAP® vial and closed with a DESICAP® cap. The closed vial is then placed in a foil bag with a desiccant, and the foil bag is then heat-sealed to form the packaged formulation.

[0062]

[0088] Stability testing

[0089] With the present drug particle formulations, it is desirable to minimize exposure to uncontrolled ambient humidity. Drug particles according to embodiments of the present invention have been shown to be stable for at least 9 months when stored under controlled humidity conditions at 25°C / 60% RH. In some embodiments, drug particles have been shown to be stable for at least 6 months when stored under controlled humidity conditions at 40°C / 75% RH. In some embodiments, drug particles have been shown to be stable for at least 9 months when stored under dry conditions at 25°C / 60% RH. In some embodiments, drug particles have been shown to be stable for at least 6 months when stored under dry conditions at 40°C / 75% RH. A study was conducted to determine the stability of drug particles at % RH.

[0063]

[0090] Prototype stability testing

[0091] The purpose of prototype stability testing was to evaluate the stability of drug particles in capsules. Both 25 μg and 75 μg strengths were evaluated when stored at 25°C / 60% RH and 40°C / 75% RH. For testing, drug particles were placed in size 3 HPMC opaque capsules (Capsugel Vcaps). Ten filled capsules were placed in HDPE vials (Desicap), which were sealed with stoppers. The stoppered vials were placed in foil wrapped with desiccant sachets.

[0064]

[0092] Data for a 25 μg dose of drug particles stored at 25° C. / 60% RH are shown in the table below.

[0065] [Table 8]

[0066]

[0093] Data for a 25 μg dose of drug particles stored at 40° C. / 75% RH are shown in the table below.

[0067] [Table 9]

[0068]

[0094] Data for a 75 μg dose of drug particles stored at 25° C. / 60% RH are shown in the table below.

[0069] [Table 10]

[0070]

[0095] Data for a 75 μg dose of drug particles stored at 40° C. / 75% RH are shown in the table below.

[0071] [Table 11]

[0072]

[0096] Stability testing of clinical trial materials

[0097] The purpose of the stability study of the clinical trial material was to evaluate the stability of drug particles in capsules. Three strengths were evaluated: 25 μg, 50 μg, and 75 μg active drug doses in capsules. As with the previous study, two storage conditions were evaluated: 25°C / 60% RH and 40°C / 75% RH. For testing, the drug particles were placed in size 3 HPMC opaque capsules (Capsugel Vcaps). Ten filled capsules were placed in HDPE vials (DESICAP), which were then sealed with stoppers. The stoppered vials were placed in foil wrapped with a desiccant sachet.

[0073]

[0098] Data for a 25 μg dose of drug particles stored at 25° C. / 60% RH are shown in the table below.

[0074] [Table 12]

[0075]

[0099] Data for a 25 μg dose of drug particles stored at 40°C / 75% RH are shown below. The table below shows the results.

[0076] [Table 13]

[0077]

[0100] The following data are for a 50 μg dose of drug particles stored at 25°C / 60% RH. As shown in the table below.

[0078] [Table 14]

[0079]

[0101] The following data are for a 50 μg dose of drug particles stored at 40°C / 75% RH. As shown in the table below.

[0080] [Table 15]

[0081]

[0102] The following data are for a 75 μg dose of drug particles stored at 25°C / 60% RH. As shown in the table below.

[0082] [Table 16]

[0083]

[0103] The following data are for a 75 μg dose of drug particles stored at 40°C / 75% RH. As shown in the table below.

[0084] [Table 17]

[0085]

[0104] Dry Powder Inhaler Device

[0105] The RS00 Model 8 is certified to Plastiape standards according to ISO and FDA standards. The RS00 Model 8 device is a commercially available single-dose dry powder inhaler device manufactured by RS00 Model 8 GmbH & Co. KG (Italy). The overall design of the RS00 Model 8 device is shown in Figure 11.

[0086]

[0106] The cap held onto the mouthpiece prevents dust and other foreign matter from entering the inhaler when not in use. The cap is designed to prevent air from entering the device. The plastic side covers the air inlet hole, but the cap does not provide an airtight seal for the device. The cap does not form part of the activation process.

[0087]

[0107] During assembly, the mouthpiece is attached to the inhaler body, but can be removed for cleaning purposes. The mouthpiece can be easily removed and attached. To assemble the mouthpiece, place the offset pegs on the base of the mouthpiece into the corresponding holes in the inhaler body and rotate the mouthpiece until it snaps shut. The snap closure ensures proper alignment of the mouthpiece and inhaler body and prevents unwanted airflow. The mouthpiece contains a mesh that reduces particle size and prevents accidental ingestion of the capsule during inhalation.

[0088]

[0108] The inhaler body contains two side buttons, each of which pierces the capsule. The inhaler contains four pins for piercing the mouthpiece. The pins are inserted into corresponding housings in the pushbutton, and the pin heads are maintained in their position by a back plate that is ultrasonically welded to the pushbutton. The buttons and pins are each maintained in their outward position by four small steel springs in each button. A three-component snap-lock system in the inhaler body ensures that the mouthpiece is properly positioned when closed.

[0089]

[0109] The capsule perforation area is located internally adjacent to the pin. When the capsule is inserted into the puncture area, the button is released and the pin on the button pierces the end of the capsule, preparing it for ejection. Above the capsule puncture area are two tangential air inlets and a circular chamber. These rotate the capsule when the patient inhales through the device. The rotation of the capsule creates a centrifugal effect on the powder, promoting efficient ejection.

[0090]

[0110] The performance of the dry powder inhaler, measured previously, was The combination of properties of the inhaler device itself, including the amount of oxygen consumed, and the type of fluid consumed.

[0111] Nonclinical trials

[0112] Treprostinil is a tricyclic benzidine analogue of endogenous PGI2. The primary pharmacological effects of treprostinil are direct vasodilation of pulmonary and systemic arterial vascular beds and inhibition of platelet aggregation. Treprostinil was developed for chronic administration by continuous subcutaneous infusion as a treatment for patients with PAH. PGI2, an endothelial cell-derived substance, is a potent vasodilator and inhibitor of platelet aggregation. The hemodynamic properties of treprostinil are similar to those of PGI2; however, unlike PGI2, treprostinil is chemically stable.

[0091]

[0113] Evaluating the pharmacokinetics (PK) and toxicity of the dry powder treprostinil formulation of the present invention To investigate this, a series of in vivo studies were performed.

[0114] A pilot, non-GLP, single-dose, inhaled trial of treprostinil in dogs K Exam (Exam 19073)

[0115] This study was conducted in three beagle dogs with a target lung deposition of 3 μg / kg (D The single-dose PK of LIQ861 (administered via a PI) was compared to Tyvaso (administered via a nebulizer). Results demonstrated a generally similar treprostinil PK profile following administration of LIQ861 compared with Tyvaso. In this pilot single-dose PK study, treprostinil (dry powder formulation; estimated lung deposition of 3.0-3.4 μg / kg) and treprostinil (nebulized liquid; target lung deposition of 3 μg / kg) were compared in three beagle dogs. Results demonstrated a generally similar treprostinil PK profile following administration of treprostinil (dry powder formulation) compared with treprostinil (nebulized liquid). The study design and results are discussed in more detail herein.

[0092]

[0116] Applicants administered the compound as a single inhalation exposure (with controlled ventilation) to anesthetized beagle dogs. A study was conducted to compare plasma concentrations and pharmacodynamics (PD) following administration of treprostinil sodium (nebulized liquid vs. a dry powder formulation similar to the present LIQ861 formulation). Treprostinil sodium was prepared as a nebulized liquid from the same DS used to prepare the dry powder formulation. The dry powder formulation was manufactured using PRINT technology and utilized the same drug substance, treprostinil sodium, but with different excipient concentrations compared to LIQ861. Importantly, the present excipient concentrations are highly consistent, providing for reproducible batch-to-batch manufacturing of the LIQ861 product. The formulations used in this study are referred to as treprostinil (nebulized liquid) and treprostinil (dry powder formulation), respectively, in the description of this study. The study design, results, and conclusions are described below.

[0093]

[0117] In Study 19073, three dogs received nebulized treprostinil (nebulized liquid Dogs received a single inhaled dose of treprostinil (dry powder formulation; estimated lung deposition of 3.0-3.4 μg / kg) after a 2-day washout period. Blood was collected for plasma analysis of treprostinil concentrations before each dose and 2, 5, 10, 20, 30, 60, 120, and 180 minutes after the end of each dose. Additionally, two different dogs (one assigned to each treprostinil formulation) were used to monitor the following PD endpoints (hemodynamic changes): systemic arterial blood pressure [mean arterial pressure (MAP, mmHg), systolic arterial pressure (mmHg), diastolic arterial pressure (mmHg)], pulmonary artery pressure (PAP, mmHg), right atrial pressure (RAP, mmHg), pulmonary capillary wedge pressure (PCWP, mmHg) or left atrial pressure (mmHg), cardiac output (CO, L / min, triplicate average), total peripheral resistance (TPR), pulmonary vascular resistance (PVR), and heart rate (HR). PD effects were assessed before the start of administration and at target times of 5, 10, 20, 30, 60, 120, and 180 minutes after the end of administration. Unlike the first three dogs, the dog assigned to monitor PD effects was anesthetized for the duration of data collection. Dogs assigned treprostinil (nebulized liquid) were administered an estimated lung deposition of 4.0 μg / kg. Dogs assigned to the (dry powder formulation) were administered an estimated lung deposition of 2.5 μg / kg. Blood was collected at the same time points as the first three dogs.

[0094]

[0118] Treprostinil (nebulized liquid and dry powder formulations) is effective in treating HR, PAP, and RA. It had no effect on P, PCWP, or CO, but had a small effect on the reduction and subsequent increase of arterial blood pressure. Treprostinil (atomized liquid) appeared to decrease stroke volume, increase TPR, and decrease PVR. Treprostinil (dry powder formulation) appeared to increase stroke volume, decrease TPR, and decrease PVR. The study administrator concluded that the pilot data were inconclusive for comparing the potential PD effects of treprostinil (atomized liquid) with those of treprostinil (dry powder formulation). However, there did not appear to be any significant differences in PD effects associated with administration of either formulation of treprostinil.

[0095]

[0119] A pilot, non-GLP, single-dose, inhalation study of LIQ861 in male rats PK study (Study 75670)

[0120] In this study, a range of LIQ861 doses were administered as a single inhalation up to a feasible dose. The PK of treprostinil was evaluated in male rats after LIQ861 administration. Systemic exposure data from this study were used to determine the appropriate dose and blood sampling time for a definitive comparative PK bridging study of LIQ861 and nebulized treprostinil. The results of this study were used to select the dose level and optimal blood sampling paradigm for the definitive PK bridging study.

[0096]

[0121] Summary: Trial 75670

[0122] The purpose of this study is to evaluate the efficacy of PRINT treprostinil dry powder ( The objective of this study was to determine the pharmacokinetic profile of treprostinil in male Sprague Dawley rats when administered as a single 4-hour inhalation of PRINT-Tre at target dose levels of 0.15, 0.75, and 1.5 mg / kg. The results of this study were used to determine appropriate dose levels and blood sampling time points for the final PK bridging study.

[0097]

[0123] The test items were administered to three male rats per group as described in the table below. The patient received a single dose by inhalation.

[0098] [Table 18]

[0099]

[0124] Mortality, clinical signs and body weight were assessed. Blood samples were taken and trephined. The rostinil content was analyzed.

[0125] No mortality occurred, no clinical signs were observed, and body weight was not affected. .

[0100]

[0126] Overall achieved gravimetric and analytical aerosols for all groups Concentrations were within 16% of the target concentration. Corresponding mean treprostinil dose levels for all groups were within 7% of the target dose levels, and clear dose differences between groups for each gender were achieved. Gravimetric particle size MMAD for all groups was 1.2 to 1.6 μm (GSD 2.06 to 2.56). For both treprostinil and trehalose, chemical measurements of particle size distribution ranged from 1.3 to 1.8 μm, with corresponding GSDs between 1.65 and 2.15. Particle size distributions were considered gravimetrically and chemically respirable.

[0101]

[0127] Non-compartmental analysis of the mean treprostinil plasma concentration dataset The mean PK parameters for the PRINT-Tre treatment group obtained using the method are summarized below.

[0102] [Table 19]

[0103]

[0128] In conclusion, 1.409 mg / kg in Sprague-Dawley rats A single 4-hour inhalation dose of PRINT-Tre at a high mean treprostinil dose of 0.05g / day was well tolerated, with no significant test-item-related findings. Exposure to treprostinil generally increased dose-proportionally between the low and mid-dose groups. Exposure increased slightly less than dose-proportionally between the mid-dose and high-dose groups. However, animals in the high-dose group were exposed to aerosol concentrations well below the target for the final 16-26 minutes of inhalation, which may explain the less-than-dose-proportional increase in exposure. Based on these results, a similar dose level is recommended for the following definitive PK study. Blood sampling time points during the test-item inhalation period could be adjusted to better characterize exposure during test-item administration.

[0104]

[0129] Introduction

[0130] The purpose of this study is to evaluate the efficacy of PRINT treprostinil dry powder ( PRINT-Tre) at target dose levels of 0.15, 0.75, and 1.5 mg / kg The objective of this study was to determine the pharmacokinetic profile of treprostinil in male Sprague Dawley rats when administered as a single 4-hour inhalation at 100 rpm. The results of this study will be used to determine appropriate dose levels and sampling time points for the final PK bridging study.

[0105]

[0131] The study was not conducted in accordance with GLP regulations, but rather with the appropriate standard operating procedures (S I followed the OP.

[0132] Experimental design

[0133] The test items were administered daily by inhalation to rats as described in the table below. It was administered to the rhesus monkey group.

[0106] [Table 20]

[0107]

[0134] After administration, six serial blood samples were collected for pharmacokinetic evaluation.

[0108] [Table 21]

[0109]

[0135] treatment

[0136] Acclimatization to the exposure system

[0137] Before the animals were exposed to the test item aerosol, the rats were Acclimatization to Restraint Procedures: Animals were gradually accustomed to being restrained in the dosing tubes used during exposure until the period used for aerosol dosing.

[0110]

[0138] Animal exposure Exposure system used: Flow-past rodent inhalation exposure system Exposure method: Inhalation via nose-only exposure Test item type: Dry powder formulation Generation method: Piston supply / Rotary brush generator Exposure time: 240 minutes

[0139] Target aerosol concentrations and dose levels were as follows:

[0111] [Table 22]

[0112]

[0140] Assessment of achieved dosage levels Target dosage levels were estimated using the following formula:

[0113]

number

[0114] D L = achieved dose level (mg / kg / day) E c = actual concentration delivered to the animal (mg / L air) RMV = Respiratory volume per minute (L / min) according to the method of Bide, Armour and Yee J. App. Toxicol., Vol. 20, 2000: RMV (L / min) = 0.499 × BW (kg) 0.809 T = time, duration of daily exposure (min) BW = average body weight (kg) during the exposure period. This assessment of the total inhaled dose assumed 100% deposition in the respiratory tract.

[0115]

[0141] Inhalation Exposure System

[0142] The powder aerosol was generated using a piston-fed / rotating brush generator. The aerosol was diluted as necessary to achieve the target aerosol concentration and placed in a 40mm diameter container. The aerosol was discharged through a tube into a flow-past inhalation exposure system. The airflow rate through the exposure system was monitored and recorded manually during each aerosol generation period. Airflow into the exposure system was adjusted by the absolute volume of air supplied to the generator using a volumetric flow meter. Adjustment of the aerosol exhaust flow from the animal exposure system was achieved using an exhaust valve, and the overall balance of airflow in the exposure system was monitored using a pressure gauge. The system provided a minimum of 1.0 L / min to each animal exposure port and was balanced to ensure a slight positive pressure at the site of proposed animal exposure. This ensured that the generated aerosol was not diluted. Delivery of equal volumes of aerosol to each proposed exposure location was achieved by using the same distribution network for each individual exposure location attached to the system.

[0116]

[0143] Inhalation System Monitoring

[0144] Determination of aerosol concentration, particle size distribution, oxygen concentration, relative humidity and temperature is performed during the exposure Measurements were performed on samples collected from representative ports in the chamber. Sample flow rates were precisely adjusted using a variable area flow meter that was calibrated prior to use using a primary airflow calibrator. Absolute aerosol concentration for each sample was measured with a wet gas meter.

[0117]

[0145] oxygen concentration

[0146] The oxygen concentration of the generated atmosphere was measured once during each aerosol exposure. The oxygen concentration in the exposure atmosphere was maintained at 19-23%.

[0118]

[0147] Relative humidity / temperature

[0148] The temperature and relative humidity of the generated atmosphere were maintained at 100°C for 1 hour during each aerosol exposure. The temperature of the exposure atmosphere was maintained at 19-24°C.

[0119]

[0149] Determination of aerosol concentration

[0150] To determine the gravimetric concentration of the test item in the generated aerosol At least one aerosol concentration filter sample was collected on a glass fiber filter and weighed each day. The filter samples were transferred to an analytical chemistry laboratory for chemical determination of treprostinil and trehalose concentrations using analytical methods (Test No. 41609 and Test No. 41635).

[0120]

[0151] Determination of particle size distribution and mass median aerodynamic diameter (MMAD)

[0152] The particle size distribution of the generated aerosol was measured by passing the sample through a seven-stage mercer cascade inlet. Measurements were taken once during each exposure by collection into a 7-Stage Mercer Cascade Impactor. MMAD and geometric standard deviation (GSD) were calculated based on the results obtained from the impactor using a log-probit transformation.

[0121]

[0153] Survival observation

[0154] mortality rate

[0155] Mortality checks were performed at least daily during all phases of the study.

[0122]

[0156] Clinical observations

[0157] Cageside clinical signs (poor health, behavioral changes, etc.) Symptoms were recorded at least once daily during all phases of the study, except on detailed clinical trial days when symptoms were replaced by DCEs.

[0123]

[0158] A detailed clinical examination of each rat is conducted as part of its health status upon arrival. The test was performed on the first day before administration.

[0159] Animals deemed healthy are further evaluated by a clinical veterinarian. was examined.

[0124]

[0160] body weight

[0161] Body weights were measured for all animals once upon arrival as a health status and before group assignment. Recorded once before and on day 1 (pre-dose).

[0125]

[0162] Pharmacokinetics

[0163] A series of six blood samples (approximately 0.3 mL each) was collected from each rat on day 1. Blood samples were taken from each rat at -15, 5, 15, 30, 75, and 105 minutes after treatment. Thus, a total blood volume of 1.8 mL was collected from each rat during the course of the study. For this purpose, blood was collected from each (unanesthetized) rat by jugular vein puncture, and samples were collected in tubes containing the anticoagulant K2EDTA. The tubes were placed on wet ice pending processing.

[0126]

[0164] After collection, the sample was centrifuged (2500 rpm, 10 minutes at approximately 4°C) Plasma was collected and stored frozen (below -60°C) in labeled tubes.

[0165] Deviations in pharmacokinetic time points were noted in the raw data and made available with the samples. The location of blood collection was noted in the raw data.

[0127]

[0166] Noncompartmental analysis of plasma treprostinil concentrations was performed using the Phoenix x Performed by using WinNonlin 6.3 software.

[0167] The following settings were used for the analysis: Sampling method: Sparse AUC calculation method: Linear trapezoidal method with linear interpolation Lambda Z(λ z ) method: λ z Best fit for logarithmic regression Weighting (λ z Calculation): Uniform

[0168] The pharmacokinetic parameters (including abbreviations and descriptions for each parameter) are listed below: Listed in the table:

[0128] [Table 23]

[0129]

[0169] Data Evaluation and Statistics

[0170] Numerical and non-numeric data obtained during testing are reported simply as individual values. was done.

[0130]

[0171] result

[0172] Aerosol concentration

[0173] The gravimetric test atmosphere concentrations achieved were as follows:

[0131] [Table 24]

[0132]

[0174] The achieved analytical test ambient concentrations for treprostinil were: Ta:

[0133] [Table 25]

[0134]

[0175] The achieved analytical test atmosphere concentrations for trehalose were as follows:

[0135] [Table 26]

[0136]

[0176] The overall achieved gravimetric and analytical aerosol concentrations were The generated atmospheres were within 16% of the target concentration, except for Group 3. All CV% were below 20% and therefore considered stable throughout the treatment period. The increase in CV% for Group 3 was caused by a malfunction of the rotating brush generator (RBG) as no test item remained in the canister with 26 minutes remaining in the generation (16 minutes after administration for Animal 3001A, 21 minutes for Animal 3002A, and 26 minutes for Animal 3003A). Although a new test item canister was installed in the RBG device, the aerosol concentration was significantly lower than targeted during the final 26 minutes. However, since there was a significant difference in aerosol concentration between groups, the overall aerosol concentration was still considered acceptable for this study.

[0137]

[0177] Dosage Levels

[0178] The overall achieved doses of treprostinil are shown below:

[0138] [Table 27]

[0139]

[0179] The mean dose levels achieved in all groups were >7% of the target dose levels. The dose levels were considered acceptable for the study because they were within the range of 0.1 to 1.0 mg / kg and therefore achieved clear dose differences between groups for each gender.

[0140]

[0180] Particle size distribution

[0181] The measured data for the mean weight particle size distribution were as follows:

[0141] [Table 28]

[0142]

[0182] The mean chemical measurements of particle size distribution for treprostinil were as follows:

[0143] [Table 29]

[0144]

[0183] The mean chemical measurements of particle size distribution for trehalose were as follows:

[0145] [Table 30]

[0146]

[0184] The MMAD was less than 4 μm and the GSD was within 1.5 and 3. The size distribution was considered respirable in this study.

[0185] Exposure chamber conditions

[0186] The exposure chamber conditions for the reported aerosol concentration exposures are summarized below:

[0147] [Table 31]

[0148]

[0187] The range of oxygen concentration, temperature and relative humidity of the exposure atmosphere must be acceptable in all cases. It was thought to be a noh play.

[0188] mortality rate

[0189] There were no deaths during this study.

[0149]

[0190] Clinical symptoms

[0191] No adverse clinical symptoms were observed during the study.

[0192] In animal 3001A just before the 15-minute time point, there was a slight decrease in activity, ciliary and Severe eye movements and partially closed eyes were observed, but these were not observed subsequently or in any other animals and were therefore not thought to be related to the test item.

[0150]

[0193] body weight

[0194] Body weight was taken for the purpose of calculating dose levels.

[0195] Pharmacokinetics

[0196] Mean C after administration of PRINT-Tre at all achieved dose levels max The mean maximum plasma concentration (T max ) was reached at 3.75 hours (15 minutes before the end of administration) in all groups. 0-Tlast (AUC INF ) is 17.320(18.335)~121.258 (137.512) time * The range was ng / mL. max After administration, treprostinil plasma concentrations were observed with an estimated mean T ranging from 1.01 to 1.68 hours. 1 / 2 gradually decreased.

[0151]

[0197] Exposure to treprostinil (C max , AUC 0-T last and AUC INF(based on ) generally increased dose-proportionally between the low dose (0.158 mg / kg) and the mid-dose (0.707 mg / kg). When the dose level increased 4.5-fold from the low to the mid-dose, C max and AUC 0-Tlast Treprostinil exposure increased 4.7-fold between the mid-dose (0.707 mg / kg) and high-dose (1.409 mg / kg) doses in a slightly less than dose-proportional manner (a 2-fold increase in dose resulted in a 1.4-fold increase in exposure (C max )~1.5x(AUC 0-Tlast However, because animals in the high-dose group were exposed to much lower aerosol concentrations than targeted during the final 16–26 min of the exposure period, it is possible that exposure levels were achieved, which may explain the less than dose-proportional increase in exposure.

[0152]

[0198] conclusion

[0199] Average dose of 1.409 mg / kg / day for Sprague-Dawley rats A single 4-hour inhalation administration of PRINT-Tre at the high treprostinil dose was well tolerated, with no significant test-item-related findings. Treprostinil exposure generally increased dose-proportionally between the low and mid-dose groups. Exposure increased slightly less than dose-proportionally between the mid-dose and high-dose groups. However, animals in the high-dose group were exposed to aerosol concentrations much lower than the target during the final 16–26 min of inhalation, which may explain the less-than-dose-proportional increase in exposure. Based on these results, a similar dose level is recommended for the following definitive PK study. Blood sampling time points during the test-item inhalation period could be adjusted to better characterize exposure during test-item administration.

[0153]

[0200] Non-GLP, single-dose study of LIQ861 and nebulized treprostinil in rats Administration, inhalation, comparative PK study (Study 75658)

[0201] This study aims to establish a bridge between the two formulations by comparing the P The K profile was evaluated and compared with treprostinil (nebulized).

[0154]

[0202] A non-GLP, single-dose inhaled pharmacokinetic study of treprostinil in rats (Study 75658) was completed by Liquidia (referred to as the definitive PK bridging study). This study compared the systemic exposure of LIQ861 to nebulized liquid treprostinil sodium. The observed systemic exposure showed no significant differences between the formulations, providing a bridge between the LIQ861 formulation and the marketed Tyvaso formulation, allowing the use of Tyvaso nonclinical toxicology studies to support the LIQ861 formulation under the 505(b)(2) pathway.

[0155]

[0203] In Study 75658, LIQ861 nebulized treprostinil sodium Systemic exposure to LIQ861 was compared in rats. LIQ861 was delivered over a 4-hour exposure period at total delivered dose levels of 0.273, 0.762, and 1.50 mg / kg body weight. Nebulized treprostinil sodium was delivered at a single dose level (0.785 mg / kg total delivered dose) for the same exposure period (4 hours) as LIQ861. Blood was collected for plasma analysis of treprostinil concentrations at 30 and 60 minutes after the start of dosing, immediately after dosing (240 minutes), and 5, 15, 30, 75, and 105 minutes after dosing was completed.

[0156]

[0204] Pharmacokinetic parameters of study 75658. Individual plasma concentrations of treprostinil were The maximum plasma concentrations were reached 0.5 to 4 hours after the start of the 4-hour exposure period. The maximum concentration (Cmax) and area under the curve (AUC) values were were similar between males and females within treatment groups. Dose-related increases in Cmax and AUC values were observed across the three LIQ861 dose groups. The relative bioavailability of LIQ861 compared with nebulized treprostinil based on the dose-normalized AUC-time curves (AUCinf) extrapolated to time infinity ranged from 1.2 to 2.2.

[0157] [Table 32]

[0158] [Table 33]

[0159]

[0205] Summary: Trial 75658

[0206] The purpose of this study is to evaluate the efficacy and safety of PRINT-Treprostinil (PRINT-Tre) To determine the pharmacokinetic (PK) profile of treprostinil in Sprague-Dawley rats when administered as nebulized treprostinil sodium at 0.15, 0.75, and 1.5 mg / kg by 4-hour inhalation; to determine the PK profile of treprostinil in Sprague-Dawley rats when administered as nebulized treprostinil sodium in solution (Tre solution) at 0.75 mg / kg by 4-hour inhalation; and to compare the PK profiles of treprostinil when administered as PRINT-Tre and Tre solution.

[0160]

[0207] The test items were administered by nasal inhalation alone for 4 hours as described in the table below. , 6 male and 6 female rats per group were dosed once:

[0161] [Table 34]

[0162]

[0208] Mortality, clinical signs, and body weight were assessed. Pharmacokinetic samples were collected and analyzed. Analysis of these samples was carried out.

[0209] No mortality occurred and no clinical signs were observed.

[0163]

[0210] The overall achieved aerosol concentrations for all groups were treprostinil For both cereals and trehalose, the gravimetrically targeted concentrations were within 10% of the targeted concentrations, except for Group 2, which significantly exceeded the target concentration (76-95%). The corresponding mean dose levels achieved for all groups were within 5% of the targeted dose levels, except for Group 2, which exceeded the target dose level by 82%. However, the dose levels were considered acceptable for this study, as clear dose differences between groups for each gender were achieved.

[0164]

[0211] The particle size MMAD of groups 2 to 4 was 1.7 to 2.0 μm by gravimetric measurement. (GSD 1.90-2.67). For both treprostinil and trehalose, the chemical particle size distribution ranged from 1.6-1.8 μm with a corresponding GSD of 1.89-2.24. The particle size MMAD for Group 1 was 0.5 μm with a corresponding GSD of 2.60. The particle size distribution was considered to be respirable.

[0165]

[0212] 0.273mg / kg, 0.762mg / kg or 1.498mg / kg When PRINT-Tre was administered at the achieved dose level, plasma exposure to treprostinil was generally similar in both sexes, but exposure was slightly lower in females than in males at the mid-dose level and slightly higher in females than in males at the high-dose level.

[0166]

[0213] AUC for both genders 0-Tlast , AUC INF and C max Value-based In the control group, plasma exposure increased more than proportionally between the low and mid-dose levels. However, between the mid-dose and high-dose levels, plasma exposure decreased more than proportionally for females and did not increase for males. Maximum mean treprostinil plasma concentrations (T max ) was at the end of inhalation in both sexes, except for low-dose males and high-dose females, and the mean T max were 1 hour and 0.5 hours after the start of inhalation, respectively.

[0167]

[0214] At low dose levels, mean treprostinil plasma concentrations were significantly higher in PRINT-Tre The mean treprostinil plasma concentrations were similar at 0.5, 1, and 4 hours after inhalation exposure, suggesting that steady state was achieved within the first 30 minutes of exposure. The same was true for females at the high dose level. However, for males at the high dose level and for both sexes at the intermediate dose level, the mean treprostinil plasma concentrations were higher at the end of inhalation than at 1 hour after inhalation. After inhalation was terminated, the treprostinil plasma concentrations gradually decreased. Taking into account the degree of individual variability, the estimated mean T 1 / 2 Values were similar at all dose levels, ranging from 0.7 to 1.8 hours in males and 0.7 to 1.0 hours in females.

[0168]

[0215] For Tre solution, when administered at 0.785 mg / kg, Plasma exposure to treprostinil was generally similar in both genders. Maximum mean treprostinil plasma concentrations (T max ) was at the end of inhalation. Mean treprostinil plasma concentrations were similar to Tre solution at 0.5, 1, or 4 hours after inhalation exposure, suggesting that steady state was achieved within the first 30 minutes of exposure. After inhalation was terminated, treprostinil plasma concentrations gradually declined, with mean T 1 / 2 Values were estimated to be 0.6 hours for males and 0.8 hours for females.

[0169]

[0216] Approximately equivalent dosage levels of PRINT-Tre and Tre solutions (0.05, ... At doses of 0.76 and 0.79 mg / kg, plasma exposure to treprostinil was greater with PRINT-Tre than with Tre solution. Specifically, mean AUC 0-Tlast was approximately twice as high (61 h*ng / mL vs. 126 h*ng / mL, respectively), and the mean C max were three times higher (16 ng / mL vs. 44 ng / mL, respectively). As expected, once treprostinil entered the systemic circulation, it was cleared from plasma at similar rates, with mean T 1 / 2 The values were 0.7 to 1.0 hours for PRINT-Tre and 0.6 to 0.8 hours for Tre solution.

[0170]

[0217] In summary, 1.498 mg / kg of Sprague-Dawley rats A single 4-hour inhaled dose of PRINT-Treprostinil at a mean high dose of 1000 mg / kg / day was well tolerated with no study-related findings. At comparable dose levels, plasma exposure to treprostinil was higher with PRINT-Tre than with Tre solution. Specifically, mean AUC 0-Tlast is about twice as high, and the average C max was three times higher. As expected, once treprostinil entered the systemic circulation, it was cleared from plasma at similar rates regardless of how it was administered.

[0171]

[0218] The objectives of the study were to: 1. Determine the treprostinil pharmacokinetic (PK) profile in Sprague-Dawley rats when administered as PRINT-Treprostinil (PRINT-Tre) at 0.15, 0.75, and 1.5 mg / kg via 4-hour inhalation. 2. Determine the treprostinil PK profile in Sprague-Dawley rats when administered as nebulized treprostinil sodium solution (Tre solution) via 4-hour inhalation of 0.75 mg / kg. 3. Compare the treprostinil PK profiles when administered as PRINT-Tre and Tre solution.

[0172]

[0219] Experimental design

[0220] Test items were administered by inhalation to groups of rats as described in the table below for 4 hours. Given:

[0173] [Table 35]

[0174]

[0221] During and after the inhalation period, a series of eight blood samples for pharmacokinetic evaluation were collected. did.

[0222] Justification for choice of route of administration, species and dosage level

[0223] The route of administration was chosen to be the intended route of human therapy.

[0175]

[0224] Rats were chosen because they are the rodent species recommended by various regulatory agencies. Background data are available, and rats were used as a test system for previous toxicity studies with treprostinil sodium solution that supported the development and approval of that product. The use of rats in the current study allowed for comparison with previous studies.

[0176]

[0225] High dosage levels for PRINT-Tre are based on technical Suitable doses were achievable based on the aerosol test (Test No. 41610).

[0226] Low and intermediate dose levels of PRINT-Tre were compared in rats. It was selected based on a previous pilot PK study (Study No. 75670).

[0177]

[0227] The dosage level for Tre solution is the mid-dose level of PRINT-Tre. were chosen to allow for direct comparison.

[0178] [Table 36]

[0179] [Table 37]

[0180]

[0228] Preparation of test items

[0229] PRINT-Tre was used as provided by the sponsor. A glove box was used for handling, aliquoting or packing the canisters. The relative humidity (RH) inside the glove box was monitored and recorded using a hygrometer and maintained below 23% RH.

[0181]

[0230] For Group 1, treprostinil sodium was dissolved in purified water and diluted to the desired formulation concentration. Representative samples (0.5 mL, duplicate) were collected to confirm the formulation concentration of treprostinil in the formulation.

[0182]

[0231] treatment

[0232] Acclimatization to the exposure system

[0233] The rats were exposed to the aerosol of the test item for three days. Acclimatization to Restraint Procedures: Animals were gradually accustomed to being restrained in the dosing tubes used during exposure until the period used for aerosol dosing.

[0183]

[0234] Animal exposure Exposure system used: Flow-past rodent inhalation exposure system Exposure method: Inhalation via nose-only exposure Test item type: solution (group 1), dry powder (groups 2-4) Generation method: spray (1st group) and piston-fed / rotating brush generator (2nd-4th groups) Exposure time: 240 minutes

[0235] Target aerosol concentrations and dose levels were as follows:

[0184] [Table 38]

[0185]

[0236] Assessment of achieved dosage levels Target dosage levels were estimated using the following formula:

[0186]

number

[0187] D L = achieved dose level (mg / kg / day) E c = actual concentration delivered to the animal (mg / L air) RMV = Respiratory Minute Volume (L / min) according to the method of Bide, Armour and Yee 2000 J. App. Toxicol., Vol. 20: RMV (L / min) = 0.499 × BW (kg) 0.809 T = time, duration of daily exposure (min) BW = average body weight (kg) during the exposure period. This assessment of the total inhaled dose assumed 100% deposition in the respiratory tract.

[0188]

[0237] Inhalation Exposure System

[0238] Powder aerosols in groups 2-4 were produced using piston-fed / rotating brush generators. The first group of liquid aerosols was generated by metering a flow of formulation into a clinical nebulizer (Sidestream). The generated aerosol was diluted as necessary to achieve the target aerosol concentration and discharged into a flow-past inhalation exposure system via a 40 mm diameter tube. The airflow rate through the exposure system was monitored and recorded manually during each aerosol generation period. The airflow into the exposure system was measured using an area flow meter. The aerosol flow rate was adjusted by the absolute volume of air supplied to the animal exposure site. Regulation of the aerosol exhaust flow from the animal exposure system was achieved using an exhaust valve, and the overall balance of airflow within the exposure system was monitored using a pressure gauge. The system provided a minimum of 1.0 L / min to each animal exposure port and was balanced to ensure a slight positive pressure at the site of animal exposure. This ensured that the aerosol generated was not diluted. Delivery of equal volumes of aerosol to each exposure location was achieved by using identical distribution networks for each individual exposure location attached to the system.

[0189]

[0239] Inhalation System Monitoring

[0240] Determination of aerosol concentration, particle size distribution, oxygen concentration, relative humidity and temperature is performed during the exposure Measurements were performed on samples collected from representative ports in the chamber. Sample flow rates were precisely adjusted using a variable area flow meter that was calibrated prior to use using a primary airflow calibrator. Absolute aerosol concentration for each sample was measured with a wet gas meter.

[0190]

[0241] oxygen concentration

[0242] The oxygen concentration of the generated atmosphere was measured once during each aerosol exposure. The oxygen concentration in the exposure atmosphere was maintained at 19-23%.

[0191]

[0243] Relative humidity / temperature

[0244] The temperature and relative humidity of the generated atmosphere were maintained at 100°C for 1 hour during each aerosol exposure. The temperature of the exposure atmosphere was maintained at 19-24°C.

[0192]

[0245] Determination of aerosol concentration

[0246] At least one aerosol concentration filter sample was collected for each aerosol generation. Groups 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 1

[0193]

[0247] Determination of particle size distribution and mass median aerodynamic diameter (MMAD)

[0248] The particle size distribution of the generated aerosol was measured by passing the sample through a seven-stage mercer cascade inlet. A single measurement was performed for Groups 1-4 by collecting the aerosolized treprostinil and trehalose in a microcentrifuge. All sample materials from Groups 2-4 were weighed gravimetrically and then transferred to the analytical chemistry laboratory for chemical determination of the particle size of the aerosolized treprostinil and trehalose. All sample materials from Group 1 were transferred to the analytical chemistry laboratory only for chemical determination of the particle size of the aerosolized treprostinil. Analysis in the analytical chemistry laboratory was performed using analytical method (Test No. 41609).

[0194]

[0249] MMAD and geometric standard deviation (GSD) were calculated using a log-probit transformation. Calculations were made based on the results obtained from the impactor.

[0250] Reporting of analysis results

[0251] Prepared an analytical report containing the results of the filter analysis and particle size distribution sample analysis All samples not used in the primary analysis or all remaining samples from the primary analysis were retained until the analyst and study director determined that no confirmatory analysis was necessary. These samples were discarded, and their nature was recorded in the raw data.

[0195]

[0252] Survival observation

[0253] mortality rate

[0254] Mortality checks were performed at least daily during all phases of the study.

[0196]

[0255] Clinical observations

[0256] Cageside clinical signs (poor health, behavioral changes, etc.) Symptoms were recorded at least once daily during all phases of the study, except on detailed clinical trial days when symptoms were replaced with DCEs.

[0197]

[0257] A detailed clinical examination of each rat is conducted as part of its health status upon arrival. The test was performed on the first day before administration.

[0258] body weight

[0259] Body weights were measured for all animals once upon arrival as a health status and before group assignment. Once before and on day 1 (before dosing).

[0198]

[0260] Pharmacokinetics

[0261] A series of eight blood samples (approximately 0.3 mL each) were collected at the exposure time as shown in the table below. Blood samples were collected 30 minutes after the start of the study, 1 hour after the start of the study, immediately after the end of exposure (IPE), and at 5, 15, 30, 75, and 105 minutes after administration. Thus, a total blood volume of 1.2 mL was collected from each rat during the course of the study. For this purpose, blood was collected from each (unanesthetized) rat by jugular vein puncture, and samples were collected in tubes containing the anticoagulant K2EDTA. The tubes were placed on wet ice pending processing.

[0199] [Table 39]

[0200]

[0262] After collection, the sample was centrifuged (2500 rpm, 10 minutes at approximately 4°C) Plasma was collected and stored frozen (below -60°C) in labeled tubes.

[0263] Deviations in pharmacokinetic time points were noted in the raw data and made available with the samples. The locations of blood collection are indicated in the raw data.

[0201]

[0264] Plasma analysis was performed and bioanalytical data were prepared for inclusion in the final report.

[0265] Calculate pharmacokinetic parameters and PRINT-Tre and Tre solutions in plasma Non-compartmental analysis of treprostinil concentrations was performed by using Phoenix WinNonlin 6.3 software.

[0202]

[0266] The following settings were used for the analysis: Sampling method: Sparse AUC calculation method: Linear trapezoidal method with linear interpolation Lambda Z(λ z ) method: λ z Best fit for logarithmic regression Weighting (λ z Calculation): Uniform

[0267] The pharmacokinetic parameters (including abbreviations and descriptions for each parameter) are listed below: As listed in the table:

[0203] [Table 40]

[0204]

[0268] Data Evaluation and Statistics

[0269] Numerical and non-numeric data obtained during testing are reported simply as individual values. was done.

[0205]

[0270] result

[0271] Formulation Analysis

[0272] The formulation concentrations for Group 1 were as follows:

[0206] [Table 41]

[0207]

[0273] The formulation concentrations in Group 1 were within 2% of the target concentration, and therefore the formulation concentrations were within the range of the study. was considered acceptable.

[0274] Aerosol concentration

[0275] The gravimetric test atmosphere concentrations achieved were as follows:

[0208] [Table 42]

[0209]

[0276] The achieved chemical test ambient concentrations for treprostinil were: Ta:

[0210] [Table 43]

[0211]

[0277] The achieved chemical test atmosphere concentrations for trehalose were as follows:

[0212] [Table 44]

[0213]

[0278] Overall, the achieved aerosol concentrations were significantly higher than those of treprostin for all groups. For both treprostinil and trehalose, the gravimetrically determined aerosol concentrations were within 10% of the target concentrations, except for Group 2, which significantly exceeded the target concentrations (76% and 95% for treprostinil and trehalose, respectively). The generated atmosphere appeared stable throughout the treatment period, except for Group 2 (CV% ∼54%). However, overall aerosol concentrations was still considered acceptable in this study because there was a significant difference in aerosol concentrations between the groups.

[0214]

[0279] Dose levels achieved

[0280] The overall achieved doses of treprostinil are shown below:

[0215] [Table 45]

[0216]

[0281] The mean dose level achieved in all groups was 82% of the target dose level. The dose levels were within 5% of the target dose levels, except for Group 2, which exceeded 1. However, the dose levels were considered acceptable for the study because clear dose differences were achieved between groups for each gender.

[0217]

[0282] Particle size distribution

[0283] The measured data for the mean weight particle size distribution were as follows:

[0218] [Table 46]

[0219]

[0284] The mean chemical measurements of particle size distribution for treprostinil were as follows:

[0220] [Table 47]

[0221]

[0285] The mean chemical measurements of particle size distribution for trehalose were as follows:

[0222] [Table 48]

[0223]

[0286] The MMAD was less than 4 μm and the GSD was within 1.5 and 3. The size distribution was considered respirable in this study.

[0287] Exposure chamber conditions

[0288] The exposure chamber conditions for the reported aerosol concentration exposures are summarized below:

[0224] [Table 49]

[0225]

[0289] The range of oxygen concentration, temperature and relative humidity of the exposure atmosphere must be acceptable in all cases. It was thought to be a noh play.

[0290] mortality rate

[0291] There were no deaths during this study.

[0226]

[0292] Clinical symptoms

[0293] No clinical symptoms were observed during this study.

[0294] body weight

[0295] Body weight was taken for the purpose of calculating dose levels.

[0227]

[0296] Pharmacokinetics

[0297] 0.273mg / kg, 0.762mg / kg or 1.498mg / kg When PRINT-Tre was administered at the achieved dose level, plasma exposure to treprostinil was generally similar in both sexes, but exposure was slightly lower in females than in males at the mid-dose level and slightly higher in females than in males at the high-dose level.

[0228]

[0298] AUC for both genders 0-Tlast , AUC INF and C max Based on the value of Based on the data, plasma exposure increased more than proportionally between the low and mid-dose levels. However, between the mid-dose and high-dose levels, plasma exposure did not increase more than proportionally in females, and exposure did not increase in males. Maximum mean treprostinil plasma concentrations (T max ) was at the end of inhalation in both sexes, except for low-dose males and high-dose females, and the mean T max were 1 hour and 0.5 hours after the start of inhalation, respectively.

[0229]

[0299] At low dose levels, mean treprostinil plasma concentrations were significantly higher in PRINT-Tre The mean treprostinil plasma concentrations were similar at 0.5, 1, or 4 hours after inhalation exposure, suggesting that steady state was achieved within the first 30 minutes of exposure. The same was true for females at the high dose level. However, for males at the high dose level and for both sexes at the intermediate dose level, the mean treprostinil plasma concentrations were higher at the end of inhalation than 1 hour after inhalation. These data are summarized below.

[0230] [Table 50]

[0231]

[0300] After inhalation was terminated, treprostinil plasma concentrations gradually decreased. Considering the degree, the estimated average T 1 / 2 Values were similar at all dose levels, ranging from 0.7 to 1.8 hours in males and 0.7 to 1.0 hours in females.

[0232]

[0301] For Tre solution, when administered at 0.785 mg / kg, Plasma exposure to nil was generally similar in both sexes.

[0302] Maximum mean treprostinil plasma concentration (T max ) was the last inhalation. Mean treprostinil plasma concentrations were similar to Tre solution at 0.5, 1, or 4 hours after inhalation exposure, suggesting that steady state was achieved within the first 30 minutes of exposure. These data are summarized below.

[0233] [Table 51]

[0234]

[0303] After inhalation was terminated, treprostinil plasma concentrations gradually decreased, with an estimated mean T1 / 2 Values were estimated to be 0.6 hours for males and 0.8 hours for females.

[0304] Approximately equal doses of PRINT-Tre and Tre solutions (0.76 each) and 0.79 mg / kg), plasma exposure to treprostinil was greater with PRINT-Tre than with Tre solution. Specifically, mean AUC 0-Tlast was approximately twice as high (61 h*ng / mL vs. 126 h*ng / mL, respectively), and the mean C max were three times higher (16 ng / mL vs. 44 ng / mL, respectively). As expected, once treprostinil entered the systemic circulation, it was cleared from plasma at similar rates, with mean T 1 / 2 The values were 0.7 to 1.0 hours for PRINT-Tre and 0.6 to 0.8 hours for Tre solution.

[0235]

[0305] conclusion

[0306] Average dose of 1.498 mg / kg / day for Sprague-Dawley rats A single 4-hour inhaled dose of PRINT treprostinil at a uniformly high dose was well tolerated, with no findings related to the study items. At comparable dose levels, plasma exposure to treprostinil was higher with PRINT-Tre than with Tre solution. Specifically, mean AUC 0-Tlast is about twice as high, and the average C maxwas three times higher. As expected, once treprostinil entered the systemic circulation, it was cleared from plasma at similar rates regardless of how it was administered.

[0236]

[0307] Non-GLP, 7-day, repeated-dose, ranging findings with LIQ861 in rats ( DRF), inhalation test (Test 75654)

[0308] Results of the completed comparative PK study will be used to assess local respiratory tract toxicity and systemic trehalose toxicity. Select the dose levels to be tested in this DRF study to evaluate rostinil toxicity. Use the results to select appropriate dose levels for a 2-week GLP repeat-dose toxicity study in rats.

[0237]

[0309] Summary: Trial 75654

[0310] The purpose of this study was to administer steroids to Sprague-Dawley rats 4 times a day for 7 days. The objective of this study was to evaluate the toxicity of the excipients comprising the test item, PRINT Treprostinil, and the control item, PRINT Placebo, when administered by nasal inhalation alone for 14 days. The results were used to facilitate the selection of dose levels for a subsequent 14-day GLP inhalation toxicity study.

[0238]

[0311] Groups of six rats (3 per sex) were treated at 4:00 a.m. daily for seven days as described in the table below. Subjects were exposed to air, PRINT placebo, or PRINT treprostinil at treprostinil dose levels of approximately 170 μg / kg, 680 μg / kg, or 1370 μg / kg by inhalation for 10 min:

[0239] [Table 52]

[0240]

[0312] The particle size MMAD of groups 2 to 5 was 1.3 to 2.0 μm (GSD1) measured gravimetrically. The mean particle size distributions ranged from 1.3 to 2.1 μm, with corresponding GSDs of 1.87 to 1.95, for both treprostinil and trehalose. No deaths occurred. No clinical signs were observed, coagulation, clinical chemistry, and urinalysis parameters were unaffected, and no relevant findings were macroscopically identified in the test items.

[0241]

[0313] Rats received PRI of 1.37 mg / kg / day or less via 4-hour inhalation for 7 days. Daily administration of NT placebo or PRINT-Tre was permitted.

[0314] Introduction

[0315] The objectives of the study were to: 1. To evaluate the toxicity of the test item PRINT Treprostinil when administered to Sprague-Dawley rats by sole nasal inhalation 4 hours per day for 7 days. 2. To evaluate the toxicity of the excipients comprising the control item PRINT placebo when administered to Sprague-Dawley rats by nasal inhalation alone, 4 hours per day for 7 days. 3. The results of this dose-ranging-finding study will determine the PRINT treprostinil dose levels for the following 14-day GLP inhalation toxicity study.

[0242]

[0316] Experimental design

[0317] overview

[0318] Test and control items were administered to groups of six rats (three per sex) in the following manner: The doses were administered by inhalation for 4 hours per day for 7 days as shown in the table below. The first day of administration was designated as day 1.

[0243] [Table 53]

[0244]

[0319] The high dosage levels for PRINT-Tre are related to the technical issues with the test items. This was an achievable dose based on the aerosol test (Test No. 41610).

[0320] Low and intermediate dose levels of PRINT-Tre were The study was selected based on a previous PK study (Study No. 75658) in

[0245]

[0321] Test and control item information

[0322] Test item effects

[0323] Treprostinil is the active ingredient in PRINT-Tre and is used to treat pulmonary arterial hypertension. It is a prostacyclin compound that has been approved as a therapeutic agent.

[0246]

[0324]

[0247] [Table 54]

[0248]

[0325]

[0249] [Table 55]

[0250]

[0326] Characterization of air controls Description: Medical Grade Air (NQ 5710-500 / 2000) Supplier:Kaeser SM-11 Air Compressor

[0327] Preparation of test and control items

[0328] PRINT-Tre and PRINT placebo were administered as provided by the sponsor. A glove box under nitrogen was used for handling, aliquoting, or packing the canisters. The relative humidity (RH) inside the glove box was monitored and recorded using a hygrometer and maintained below 23% RH.

[0251]

[0329] treatment

[0330] Acclimatization to the exposure system

[0331] Before the rats were exposed to the exposure atmosphere, they were subjected to restraint procedures for 3 days. The animals were gradually habituated to being restrained in the dosing tubes used during exposure until the period used for aerosol dosing.

[0252]

[0332] Animal exposure Exposure system used: Flow-past rodent inhalation exposure system Exposure method: Inhalation via nose-only exposure Test and control item types: air (group 1), dry powder (groups 2-5) Generation method: Piston supply / Rotary brush generator (2nd to 5th groups) Exposure time: 240 minutes

[0333] Target aerosol concentrations and dose levels were as follows:

[0253] [Table 56]

[0254]

[0334] Assessment of achieved dosage levels Target dosage levels were estimated using the following formula:

[0255]

number

[0256] D L = achieved dose level (mg / kg / day) E c = actual concentration delivered to the animal (mg / L air) RMV = Respiratory volume per minute (L / min) according to the method of Bide, Armour and Yee. J. App. Toxicol., Vol. 20, 2000: RMV (L / min) = 0.499 × BW (kg) 0.809 T = time, duration of daily exposure (min) BW = average body weight (kg) during the exposure period. This assessment of the total inhaled dose assumed 100% deposition in the respiratory tract.

[0257]

[0335] Inhalation Exposure System

[0336] Powder aerosols in groups 2-5 were produced using piston-fed / rotating brush generators. The generated aerosol was diluted as necessary to achieve the target aerosol concentration and released into a flow-past inhalation exposure system via a 40 mm diameter tube. The airflow rate through the exposure system was monitored and recorded manually during each aerosol generation period. Airflow into the exposure system was adjusted by the absolute volume of air supplied to the generator using a volumetric flow meter. Regulation of the aerosol exhaust flow from the animal exposure system was achieved using an exhaust valve, and the overall balance of airflow in the exposure system was monitored using a pressure gauge. The system provided a minimum of 1.0 L / min to each animal exposure port and was balanced to ensure a slight positive pressure at the site of animal exposure. This ensured that the generated aerosol was not diluted. Delivery of equal aerosol volumes to each exposure location was achieved by using the same distribution network for each individual exposure location attached to the system.

[0258]

[0337] Inhalation System Monitoring

[0338] Determination of aerosol concentration, particle size distribution, oxygen concentration, relative humidity and temperature was performed using 1L / The aerosol concentration was measured for samples collected from a representative port in the exposure chamber at a collection sample flow rate of 1000 rpm. The sample flow rate was precisely adjusted using a variable area flow meter that was calibrated prior to use using a primary airflow calibrator. The absolute amount of each aerosol concentration sample was measured with a wet gas meter.

[0259]

[0339] oxygen concentration

[0340] The oxygen concentration of the generated atmosphere was measured once during each aerosol exposure. The oxygen concentration in the exposure atmosphere was maintained at 19-23%.

[0260]

[0341] Relative humidity / temperature

[0342] The temperature and relative humidity of the generated atmosphere were maintained at 100°C for 1 hour during each aerosol exposure. The temperature of the exposure atmosphere was maintained at 19-24°C.

[0261]

[0343] Aerosol concentration measurement

[0344] At least one aerosol concentration filter sample was collected for each aerosol generation. Groups 1 through 5 were collected. Groups 3 through 5 of the filter samples were weighed to determine the gravimetric concentration of the test item in the generated aerosol. The filter samples were transferred to an analytical chemistry laboratory where the treprostinil and trehalose concentrations were chemically determined. Group 2 of the filter samples was weighed to determine the gravimetric concentration of the control item in the generated aerosol. The filter samples were transferred to an analytical chemistry laboratory for the chemical determination of the trehalose concentration and to confirm the absence of treprostinil. Group 1 of the filter samples was not weighed gravimetrically and was transferred to the analytical laboratory only to confirm the absence of treprostinil and trehalose. Analysis in the analytical laboratory was performed using analytical methods (Test Nos. 41609 and 41635).

[0262]

[0345] Aerosol uniformity measurements

[0346] At least once during the test, the atmospheric uniformity of the exposure system was checked by comparing exposure groups 2 through 5. The system was tested by collecting multiple aerosol samples from the top, middle and bottom layers.

[0263]

[0347] Determination of particle size distribution and mass median aerodynamic diameter (MMAD)

[0348] The particle size distribution of the generated aerosol was measured by passing the sample through a seven-stage mercer cascade inlet. Groups 2 through 5 were measured at least once by collecting the aerosolized treprostinil and trehalose in a microcentrifuge. All sample materials from groups 3 through 5 were weighed gravimetrically and then transferred to the analytical chemistry laboratory for chemical measurement of the particle size of the aerosolized treprostinil and trehalose. All sample materials from group 2 were weighed gravimetrically and then transferred to the analytical laboratory for measurement of the particle size of the aerosolized trehalose. Analysis in the analytical laboratory was performed using analytical methods (Test Nos. 41609 and 41635).

[0264]

[0349] MMAD and geometric standard deviation (GSD) were calculated using a log-probit transformation. Calculations were made based on the results obtained from the impactor.

[0350] Reporting of analysis results

[0351] Prepared an analytical report containing the results of the filter analysis and particle size distribution sample analysis All samples not used in the primary analysis or remaining samples from the primary analysis were retained until the analyst and study director determined that confirmatory analysis was not necessary. These samples were then discarded, and their nature was recorded in the raw data.

[0265]

[0352] Standard Operating Procedures

[0353] All procedures were performed according to standard operating procedures and kept on file. Deviations from standard operating procedures were documented in the raw data.

[0266]

[0354] result

[0355] Inhalation System Monitoring

[0356] Oxygen concentration, temperature and relative humidity

[0357] The exposure chamber conditions for the reported aerosol concentration exposures are summarized below:

[0267] [Table 57]

[0268]

[0358] The oxygen concentration, temperature, and relative humidity of the exposure atmosphere are acceptable throughout the test. It was thought that...

[0359] Aerosol concentration

[0360] The gravimetric test atmosphere concentrations achieved were as follows:

[0269] [Table 58]

[0270]

[0361] The achieved test ambient concentrations for treprostinil were as follows:

[0271] [Table 59]

[0272]

[0362] The achieved test atmosphere concentrations for trehalose were as follows:

[0273] [Table 60]

[0274]

[0363] The overall achieved aerosol concentration for all groups was 2.5 times higher than the target concentration. With the exception of Group 3, which had a 3.5% higher trehalose concentration, all gravimetric concentrations were within 20% of the target concentration, and both treprostinil and trehalose were within 20%. Because inadequate target gravimetric concentrations were applied during the first two days of administration, the generated atmospheres were considered stable over the treatment period, even though all CV% were higher than 20%. Because there were significant differences in aerosol concentrations between groups, the overall aerosol concentrations were still considered acceptable in this study.

[0275]

[0364] Aerosol uniformity

[0365] The uniform concentrations of the gravimetric test atmosphere achieved were as follows:

[0276] [Table 61]

[0277]

[0366] The achieved test atmosphere uniform concentrations for treprostinil were as follows: Ta:

[0278] [Table 62]

[0279]

[0367] The achieved test atmosphere uniform concentrations for trehalose were as follows:

[0280] [Table 63]

[0281]

[0368] The chamber uniformity of the aerosol concentration is determined by the coefficient of variation of the aerosol concentration between samples being 2 This was below 0% and was therefore considered acceptable.

[0369] Particle size distribution

[0370] The measured data for the mean weight particle size distribution were as follows:

[0282] [Table 64]

[0283]

[0371] The chemical determination of particle size distribution for treprostinil was as follows:

[0284] [Table 65]

[0285]

[0372] The chemical determination of particle size distribution for trehalose was as follows:

[0286] [Table 66]

[0287]

[0373] All MMADs were less than 4 μm and GSDs were within 1.5 and 3. Therefore, the particle size distribution was considered to be respirable in this study.

[0374] Assessment of achieved dosage levels

[0375] The total achieved doses for treprostinil are shown below:

[0288] [Table 67]

[0289]

[0376] The total achieved doses for trehalose are as follows:

[0290] [Table 68]

[0291]

[0377] The total achieved doses for leucine are as follows:

[0292] [Table 69]

[0293]

[0378] The mean achieved dose levels for all groups were 2.5 times the target dose level. All were within 20% of the target dose level, except for group 3 for trehalose, which was above 6%. However, the dose levels were considered acceptable for the study, as clear dose differences were achieved between groups for each gender.

[0294]

[0379] mortality rate

[0380] There were no deaths during this study.

[0381] Clinical symptoms

[0382] No clinical symptoms were observed during the study.

[0295]

[0383] body weight

[0384] Weight or body mass potentially related to the administration of the test or control item The only difference in weight gain was slightly less in males given PRINT-Tre at 0.68 mg / kg / day and in both sexes given PRINT-Tre at 1.37 mg / kg / day compared to the air control group. The pattern of differences implies that the active ingredient, treprostinil, is responsible, not one of the excipients in PRINT-Tre.

[0296]

[0385] These data are summarized in the table below and highlight potential side effects associated with treprostinil. Important differences are in bold.

[0297] [Table 70]

[0298]

[0386] The remaining differences were considered to be incidental and not biologically important.

[0387] hematology

[0388] Mean hematology parameters potentially related to administration of the test or control item The only difference in parameters was a higher mean reticulocyte count in all groups given PRINT-Tre compared with the air control group. The magnitude of the difference was related to dose but was statistically significant only in males. The pattern of differences implies a role for the active ingredient, treprostinil, in PRINT-Tre, rather than one of the excipients.

[0299]

[0389] These data are summarized in the table below and provide insight into the potential side effects associated with treprostinil. Significant differences are shown in bold.

[0300] [Table 71]

[0301]

[0390] An increase in reticulocyte count is an appropriate response to an increased demand for RBCs. In the study, the increased reticulocyte count was not associated with differences in circulating erythrocyte mass (i.e., no differences in RBC count, hemoglobin concentration, or hematocrit), suggesting that the increased release of reticulocytes was accompanied by, and perhaps in response to, an increased rate of RBC loss, and that the erythropoietic response was sufficient to maintain normal circulating RBC counts.

[0302]

[0391] Any remaining differences between mean hematological parameters are likely to be random and not biologically significant. It was thought that

[0392] coagulation

[0393] Mean coagulation parameters considered to be related to the administration of the test or control item There were no differences in the meter. Any differences were considered to be incidental and not biologically significant.

[0303]

[0394] clinical chemistry

[0395] Mean clinical chemistry considered to be related to the administration of the test or control item There were no differences in parameters, and any differences were considered to be incidental and biologically insignificant.

[0304]

[0396] urine analysis

[0397] Urine analysis parameters that may be related to the administration of the test or control items There were no differences in the data. Any differences were considered to be random and biologically insignificant.

[0305]

[0398] Organ weight

[0399] Differences in mean organ weights potentially related to administration of the test or control item was noted in the lungs, adrenal glands, thymus, and testes.

[0306]

[0400] The remaining differences in mean organ weights were considered incidental and biologically insignificant. Ta.

[0401] lung

[0402] The mean lung / tracheal weights (absolute weights and weights relative to body weight) were 0.01 and 0.02 compared with the air control group. Compared with PRINT-Tre, lung weights increased in all groups receiving the test or control item. The difference with PRINT-Tre was greater than with PRINT placebo, and the difference was related to the PRINT-Tre dose. This pattern suggests that administration of the vehicle (presumably trehalose) resulted in a small increase (15%-17%) in lung weights, which was exacerbated by coadministration of treprostinil, as lung weights in the PRINT-Tre group increased compared with those in the PRINT placebo group.

[0307]

[0403] There were histopathological findings in the lungs that could explain the increased lung weight. Specifically, alveolar macrophages with basophilic vacuolated cytoplasm were increased in the lungs of all rats receiving PRINT placebo or PRINT-Tre at 0.68 mg / kg / day or higher. However, neither the distribution of this histopathological finding across groups nor the grade of the finding correlated well with the difference in mean lung weight, suggesting that some other factors were responsible. Because lungs were weighed before fixation, it is possible that some substances responsible for the weight gain were removed during tissue fixation and processing.

[0308]

[0404] Lung weight data are summarized in the table below and are for PRINT placebo and PRINT Differences potentially relevant to -Tre are in bold.

[0309] [Table 72]

[0310]

[0405] thymus gland

[0406] Mean thymus weights (absolute weights and weights relative to body weight) were measured using the PRINT-Tr In both sexes receiving PRINT-Tre at 0.68 mg / kg / day and in both sexes receiving PRINT-Tre at 1.37 mg / kg / day, thymus weight was slightly (although not statistically significantly) reduced compared with the air control. The pattern of differences implies a role for the active ingredient, treprostinil, in PRINT-Tre, rather than one of the excipients, as differences were also observed between the PRINT-Tre and PRINT-placebo groups. The reduction in thymus weight was not associated with a decrease in lymphocyte count or any histopathological findings.

[0311]

[0407] A decrease in thymus mass is a common symptom of nonspecific physiological or psychological stress. (Everds et al., 2013). This finding was associated with reduced weight gain (growth) and sometimes increased adrenal weight, so it was most likely secondary to stress and not a direct effect of treprostinil.

[0312]

[0408] Thymus weight data are summarized in the table below and potentially related to treprostinil. The differences are in bold.

[0313] [Table 73]

[0314]

[0409] Adrenal glands

[0410] Mean adrenal weights (absolute weights and weights relative to body weight) were measured using the PRINT-Tr There was an increase (although not statistically significant) in males receiving treprostinil at 0.17 mg / kg / day and in both sexes receiving PRINT-Tre at 1.37 mg / kg / day compared with air controls. The difference may be the result of chance and small group size (3 animals / sex), but the pattern of differences is likely related to the administration of treprostinil, at least at the high dose level, since differences were also seen between the high-dose PRINT-Tre and PRINT placebo groups. The increase in adrenal weight was not associated with histopathological findings.

[0315]

[0411] Increased adrenal mass is a common symptom of nonspecific physiological or psychological stress. (Everds et al., 2013). This finding was most likely secondary to stress and not a direct effect of treprostinil, as it was associated with reduced body weight gain (growth) and reduced thymus weight at the high dose levels.

[0316]

[0412] Adrenal weight data are summarized in the table below and are potentially related to treprostinil. The differences are in bold.

[0317] [Table 74]

[0318]

[0413] testes

[0414] In the group receiving PRINT-Tre at 0.68 mg / kg / day or more, There was a trend toward a slight decrease in testis weight (absolute weight and weight relative to body weight) compared with the air control group. The difference may be due to chance and the small group size (3 animals / sex), but the pattern of differences is likely related to treprostinil administration, as differences were also observed between the mid-dose and high-dose PRINT-Tre groups and the PRINT placebo group. The slight decrease in testis weight was not associated with any histopathological findings.

[0319]

[0415] Testicular weight data are summarized in the table below and are potentially related to treprostinil. The differences are in bold.

[0320] [Table 75]

[0321]

[0416] macroscopic findings

[0417] There was no evidence of macroscopic findings related to the test items at necropsy.

[0418] All findings were not dose-related, had low incidence, or were associated with air-controlled, placebo-controlled, or placebo-controlled It occurred in both sebo-control and treated animals and was therefore considered incidental.

[0322]

[0419] Microscopic findings

[0420] Treatment-related findings were observed in the lungs, anterior nasal cavity, and nasopharynx. Mirror findings were considered incidental or treatment-related.

[0323]

[0421] lung

[0422] In the lungs, PRINT placebo or PRINT-Tre 0.68 mg / A minimal to mild increase in alveolar macrophages with basophilic vacuolated cytoplasm was observed in all rats given ≥ 1 kg / day. This pattern of findings across groups indicates a response to the excipient (possibly trehalose). There were no associated inflammatory changes in the lung. Increased alveolar macrophages are a common finding in inhalation toxicity studies with powders. This reflects normal pulmonary clearance of inhaled particles and is not considered harmful.

[0324]

[0423] These data are summarized in the table below and are potentially relevant for test items or subjects. The differences relevant to the referenced items are in bold.

[0325] [Table 76]

[0326]

[0424] Nasal cavity and nasopharynx

[0425] In all groups receiving PRINT placebo or PRINT-Tre Goblet cell hypertrophy / hyperplasia was observed in the cranial portion of the nasal cavity and nasopharynx in at least one rat, with the incidence higher in groups receiving PRINT-Tre at 0.68 mg / kg / day or higher, and the mean grade higher in the group receiving PRINT-Tre at 1.37 mg / kg / day. This pattern suggests that administration of the excipient (possibly trehalose) sometimes results in goblet cell changes that are exacerbated by coadministration of treprostinil at high dose levels.

[0327]

[0426] Goblet cell hypertrophy / hyperplasia in the anterior nasal cavity and nasopharynx exposed to irritating compounds It is one of the most frequently observed lesions in rodents. This finding is generally considered a nonspecific protective or adaptive response and is not harmful.

[0328]

[0427] These data are summarized in the table below and are not shown for test or control items. Differences potentially relevant to the model are in bold.

[0329] [Table 77]

[0330]

[0428] Discussion and Conclusions

[0429] Rats received daily treatment with PRINT placebo or PRINT-Tre for 7 days. The dose was tolerated at 1.37 mg / kg / day or less via 4-hour inhalation.

[0331]

[0430] The only findings potentially related to the administration of the excipient (possibly trehalose) were: It was as follows: Alveolar macrophages with basophilic vacuolated cytoplasm were increased in all rats receiving PRINT placebo or PRINT-Tre at doses of 0.68 mg / kg / day or higher, i.e., in rats receiving trehalose at doses of 134 mg / kg / day or higher. The mean grade of this finding increased with trehalose dose level. This finding was not associated with inflammatory changes in the lungs and was considered to reflect normal lung clearance of inhaled particles. It was not considered to be adverse. Mean lung weights increased in groups receiving PRINT placebo or PRINT-Tre. The weight differences were independent of trehalose dose level. Instead, they were heavier in PRINT-Tre than in PRINT placebo and related to PRINT-Tre dose. This pattern suggests that administration of the vehicle (possibly trehalose) resulted in a small increase (15%-17%) in lung weights that was exacerbated by coadministration of treprostinil. Of note, the difference in lung weights across groups was The pattern of increase in alveolar macrophages across groups differed from that observed in the increase in alveolar macrophages, suggesting that the weight differences were not the result of an increase in macrophages. Histopathological findings consistent with increased lung weight were not present in the lungs. Because lungs were weighed before fixation, it is possible that some substances responsible for the increased weight were removed during tissue fixation and processing. Minimal goblet cell hypertrophy / hyperplasia in the cranial portion of the nasal cavity in at least one rat in all groups receiving PRINT placebo or PRINT-Tre. The occurrence of this finding was independent of trehalose dose level. Instead, the occurrence was more frequent with PRINT-Tre at 0.68 mg / kg / day or higher, and the mean grade was higher with PRINT-Tre at 1.37 mg / kg / day. This pattern suggests that administration of the excipient (possibly trehalose) occasionally results in goblet cell changes that are exacerbated by coadministration of treprostinil at higher dose levels. Goblet cell hypertrophy / hyperplasia is considered a nonspecific protective or adaptive response and is not adverse.

[0332]

[0431] In addition to differences in lung weight and worsening goblet cell hypertrophy / hyperplasia in the nasal cavity and nasopharynx Other findings potentially related to the administration of treprostinil as PRINT-Tre include: Slightly less growth (weight gain) in males at 0.68 mg / kg / day and in both sexes at 1.37 mg / kg / day. Mean reticulocyte counts increased at all dose levels, with the magnitude of the differences increasing with dose level. This was not considered deleterious in itself, but likely reflects an appropriate adaptive response to increased rates of RBC loss or turnover. Mean adrenal weights increased in males at 0.17 mg / kg / day, mean thymus weights decreased in both sexes at 0.68 mg / kg / day, and mean adrenal weights increased and mean thymus weights decreased in both sexes at 1.37 mg / kg / day. There were no associated differences in lymphocyte counts or histopathological findings in any organ. These organ weight differences most likely reflected stress and not a direct effect of treprostinil.

[0333]

[0432] Based on these results, an upcoming 14-day GLP inhalation toxicity study in rats is underway. It is recommended that FDA target dose levels comparable to those used in the current study.

[0433] Clinical Trial: LIQ861

[0434] Pharmacokinetics (PK) and safety were evaluated in healthy male and female volunteers. A randomized, placebo-controlled, single ascending dose study to evaluate

[0435] Clinical trials are being conducted to (1) determine the safety and tolerability of single doses; (2) Conducted to evaluate the single-dose pharmacokinetics of the particles of the present invention when administered to healthy male and female subjects.

[0334]

[0436] Six cohorts: 25, 50, 75, 100, 125, and 150 μm each In each cohort, eight subjects were randomly assigned in a 3:1 blinded ratio to receive a single dose of either particles of the invention (N=6) or placebo particles (N=2).

[0335]

[0437] PK evaluation blood was performed at T=0, 5, 10, 15, 20, 25, 30, 45, 60 , 90 and 120 minutes and 3, 4, 6 and 8 hours after administration.

[0438] Cohort 1

[0439] Eight subjects were enrolled and dosed in Cohort 1. Six subjects were on active treatment. One patient received the active treatment, and two received placebo. The active treatment was administered via dry powder inhalation (DPI) as a single capsule of 25 μg treprostinil strength, and the placebo treatment was administered via DPI as a single capsule of placebo formulation. All inhalations were administered using an RS00 inhaler.

[0336]

[0440] Blood samples were taken at pre-dose and post-inhalation nominal concentrations of 0.083, 0.167, 0.25, Blood samples were collected at 0.33, 0.417, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, and 8 hours. Analysis of plasma concentration versus time data to calculate standard pharmacokinetic (PK) parameters after inhalation was performed using Phoenix WinNonlin version 6.3 with scheduled blood sampling times. Plasma concentrations were provided only for subjects on the active dose, and the number of randomized subjects was assigned by the bioanalytical laboratory to maintain study blinding.

[0337]

[0441] The table shown in Figure 3A describes the six active subjects in Cohort 1. A summary of treprostinil concentration-time data for individual subjects, along with statistics, is included. Pretest noncompartmental PK parameters for treprostinil are summarized in the table shown in Figure 3B. Peak concentrations in three of six subjects occurred 0.33 hours after inhalation; one subject had Tmax of 0.167, 0.25, and 0.417 hours after administration, respectively. Concentrations then decayed in a monophasic profile, as shown by the log-linear plot. At 2 hours after inhalation, two of the six active subjects had measurable treprostinil concentrations, and at 2.5 and 3 hours after inhalation, only one subject had measurable concentrations. No subjects had quantifiable concentrations after the 3-hour time point.

[0338]

[0442] The mean Cmax was 0.364 ng / mL, and the most common Tmax was 0. The mean AUC value was 0.301 h*ng / mL, with a CV% of 30.2%. The mean apparent volume of distribution (Vz / F) was 68.1 L. The mean oral clearance (CL / F) was 91.0 L / h, with a range of 59.1 to 150. The variability of the CL / F values was 35.8% with a CV% of 35.8%.

[0339]

[0443] Cohort 2

[0444] Nine subjects were enrolled and dosed in Cohort 2. At least six subjects were All patients received active treatment and at least two received placebo; one subject withdrew and was replaced before the 2-hour PK sample. Subjects on shortened sampling schedules were excluded in this interim analysis. Active treatment was administered via dry powder inhalation (DPI) as a single capsule of 50 μg treprostinil strength, and placebo treatment was administered via DPI as a single capsule of placebo formulation. All inhalations were administered using an RS00 inhaler.

[0340]

[0445] Blood samples were taken at pre-dose and post-inhalation nominal concentrations of 0.083, 0.167, 0.25, Blood samples were collected at 0.33, 0.417, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, and 8 hours. Analysis of plasma concentration versus time data to calculate standard pharmacokinetic (PK) parameters after inhalation was performed using Phoenix WinNonlin version 6.3 with scheduled blood sampling times. Plasma concentrations were provided only for subjects on the active dose, and the number of randomized subjects was assigned by the bioanalytical laboratory to maintain study blinding.

[0341]

[0446] The table shown in Figure 4A describes the six active subjects in Cohort 2. Summary of treprostinil concentration-time data for individual subjects along with statistics are included.

[0447] The peak concentration in four of the six subjects occurred 0.17 hours after inhalation; One subject had a Tmax of 0.083 hours post-dose and one subject had a Tmax of 0.417 hours post-dose. At 2.5 hours post-dose, two of the six active subjects had measurable treprostinil concentrations, and only one subject had a measurable concentration at 3 hours post-dose. No subjects had quantifiable concentrations after the 3-hour time point.

[0342]

[0448] Pre-trial non-compartmental PK parameters of treprostinil for cohort 2 The parameters are summarized in the table shown in Figure 4B. The mean Cmax was 0.572 ng / mL, and the most frequent Tmax was 0.167 hours after inhalation. The mean AUCinf value was 0.422 h*ng / mL, with a CV% of 62.8%. The mean apparent volume of distribution (Vz / F) was 110 L. The mean oral clearance (CL / F) was 208 L / h, with a range of 67 to 624. The variability of the CL / F values was 101.5% CV%.

[0343]

[0449] In comparison, the mean Cmax in Cohort 1 was 0.364 ng / mL, and the AU Cinf values averaged 0.301 h*ng / mL. Thus, doubling the treprostinil dose resulted in an approximately 50% increase in exposure. Vz / F and CL / F values were significantly higher and more variable in Cohort 2.

[0344]

[0450] Cohort 3

[0451] Eight subjects were enrolled and administered in Cohort 3. Six subjects received active treatment. One patient received treprostinil and two received placebo. Active treatment was administered via dry powder inhalation (DPI) as a single capsule of 75 μg treprostinil strength, and placebo treatment was administered via DPI as a single capsule of placebo formulation. All inhalations were administered using an RS00 inhaler.

[0345]

[0452] Blood samples were taken at pre-dose and post-inhalation nominal concentrations of 0.083, 0.167, 0.25, Blood samples were collected at 0.33, 0.417, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, and 8 hours. Analysis of plasma concentration versus time data to calculate standard pharmacokinetic (PK) parameters after inhalation was performed using Phoenix WinNonlin version 6.3 with scheduled blood sampling times. Plasma concentrations were provided only for subjects on the active dose, and the number of randomized subjects was assigned by the bioanalytical laboratory to maintain study blinding.

[0346]

[0453] The table shown in Figure 5A describes the six active subjects in Cohort 3. Summary of treprostinil concentration-time data for individual subjects along with statistics are included.

[0454] The highest concentrations in 3 of the 6 subjects occurred 0.25 hours after inhalation; One subject had a Tmax of 0.083 hours post-dose, one had a Tmax of 0.17 hours post-dose, and one had a Tmax of 0.417 hours post-dose. At 3 hours post-dose, two of the six active subjects had measurable concentrations of treprostinil. No subjects had quantifiable concentrations after the 3-hour time point.

[0347]

[0455] Pre-trial non-compartmental PK parameters of treprostinil for Cohort 3 The parameters are summarized in the table shown in Figure 5B. The mean Cmax was 0.728 ng / mL, and the most frequent Tmax was 0.25 hours after inhalation. The mean AUCinf value was 0.757 h*ng / mL, with a CV% of 39.4%. The mean apparent volume of distribution (Vz / F) was 97 L. The mean oral clearance (CL / F) was 112 L / h, with a range of 58 to 161. The variability of the CL / F values was 39.4% CV%.

[0348]

[0456] In comparison, the Cmax in Cohort 1 and Cohort 2 was an average of 0.36 The Cmax and AUCinf values were 4 ng / mL and 572 ng / mL, respectively, with mean AUCinf values of 0.301 h*ng / mL and 0.422 h*ng / mL. Thus, tripling the dose from Cohort 1 resulted in an approximately 100-150% increase in exposure. CL / F values in Cohort 3 were more consistent with and had similar variability to those observed in Cohort 1 than those observed in Cohort 2. These results indicate that both Cmax and AUCinf can increase proportionally with increasing dose, and that CL is independent of dose over the range of 25-75 μg treprostinil.

[0349]

[0457] Cohort 4

[0458] Eight subjects were enrolled and dosed in Cohort 4. Six subjects were on active treatment. One patient received the active treatment, and two received placebo. Active treatment with 100 μg treprostinil was administered via dry powder inhalation (DPI) as two capsules of 50 μg treprostinil strength, and placebo treatment was administered via DPI as two capsules of a placebo formulation. All inhalations were administered using an RS00 inhaler.

[0350]

[0459] Blood samples were taken at pre-dose and post-inhalation nominal concentrations of 0.083, 0.167, 0.25, Blood samples were collected at 0.33, 0.417, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, and 8 hours. Analysis of plasma concentration versus time data to calculate standard pharmacokinetic (PK) parameters after inhalation was performed using Phoenix WinNonlin version 6.3 with scheduled blood sampling times. Plasma concentrations were provided only for subjects on the active dose, and the number of randomized subjects was assigned by the bioanalytical laboratory to maintain study blinding.

[0351]

[0460] The table shown in Figure 6A describes the six active subjects in Cohort 4. Summary treprostinil concentration-time data for individual subjects is included along with statistics. Peak concentrations in two of six subjects occurred at 0.25 hours after inhalation; of the remaining subjects, two had Tmax at 0.5 hours, one at 0.17 hours, and one at 0.33 hours. At 4 hours after inhalation, three of the six active subjects had measurable concentrations of treprostinil. No subjects had quantifiable concentrations after the 6 or 8 hour time points.

[0352]

[0461] Preliminary non-compartmental PK parameters of treprostinil for Cohort 4 The parameters are summarized in the table shown in Figure 6B. The mean Cmax was 1.08 ng / mL, with the most frequent Tmax values observed at 0.25 and 0.5 hours after inhalation. The mean AUCinf value was 1.22 ng / mL, with a CV% of 18.4%. The mean apparent volume of distribution (Vz / F) was 96 L. The mean oral clearance (CL / F) was 84.8 L / h, with a range of 68.3 to 122. The variability (CV%) of the CL / F values was 22.8%.

[0353]

[0462] In comparison, the Cmax for cohorts 1, 2, and 3 was an average of 0.364 nM each. The Cmax and AUCinf values were 0.301 h*ng / mL, 0.572 ng / mL, and 0.728 ng / mL, respectively, with mean AUCinf values of 0.301 h*ng / mL, 0.422 h*ng / mL, and 0.757 h*ng / mL. Thus, quadrupling the dose from cohort 1 resulted in an approximately 200-300% increase in exposure, and doubling the dose from cohort 2 resulted in an approximately 2-fold increase in exposure. These results indicate that both Cmax and AUCinf can increase proportionally with increasing dose, and that CL / F is independent of dose over the range of 25-100 μg treprostinil.

[0354]

[0463] Cohort 5

[0464] Eight subjects were enrolled and dosed in Cohort 5. Six subjects were on active treatment. Two patients received the active treatment, and two received placebo. Active treatment with 125 μg treprostinil was administered via dry powder inhalation (DPI) as one capsule of 75 μg treprostinil strength and one capsule of 50 μg treprostinil strength, and placebo treatment was administered via DPI as two capsules of placebo formulation. All inhalations were administered using an RS00 inhaler.

[0355]

[0465] Blood samples were taken at pre-dose and post-inhalation nominal concentrations of 0.083, 0.167, 0.25, 0.33, 0.417, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, Blood samples were collected at 6 and 8 hours. Analysis of plasma concentration versus time data to calculate standard pharmacokinetic (PK) parameters after inhalation was performed using Phoenix WinNonlin version 6.3 with scheduled blood sampling times. Plasma concentrations were provided only for subjects on the active dose, and the number of randomized subjects was assigned by the bioanalytical laboratory to maintain study blinding.

[0356]

[0466] The table shown in Figure 7A describes the six active subjects in Cohort 5. Summary of treprostinil concentration-time data for individual subjects with statistics is included. Peak concentrations in three of six subjects occurred at 0.17 hours after inhalation; of the remaining subjects, two had Tmax at 0.33 hours and one at 0.42 hours after administration. At 3.5 and 4 hours after inhalation, only one of six active subjects had measurable concentrations of treprostinil. No subjects had quantifiable concentrations after the 6 or 8 hour time points.

[0357]

[0467] Pre-trial non-compartmental PK parameters of treprostinil for cohort 5 The parameters are summarized in the table shown in Figure 7B. The mean Cmax was 1.19 ng / mL, with the most frequent Tmax value observed 0.17 hours after inhalation. The mean AUCinf value was 1.15 h*ng / mL, with a CV% of 44.9%. The mean apparent volume of distribution (Vz / F) was 101 L. The mean oral clearance (CL / F) was 141 L / h, with a range of 65.7 to 336. The variability (CV%) of the CL / F values was 69.9%.

[0358]

[0468] In comparison, the Cmax for cohorts 1, 2, 3, and 4 was an average of 0.36 The Cmax and AUCinf values were 0.4 ng / mL, 0.572 ng / mL, 0.728 ng / mL, and 1.08 ng / mL, respectively, and the mean AUCinf values were 0.301 h*ng / mL, 0.422 h*ng / mL, 0.757 h*ng / mL, and 1.22 h*ng / mL. Thus, a five-fold increase in dose from Cohort 1 resulted in an approximately 220-280% increase in exposure. These results indicate that both Cmax and AUCinf can increase proportionally with increasing dose, and that CL / F is independent of dose over the range of 25-125 μg treprostinil.

[0359]

[0469] Cohort 6

[0470] Cohort 6 was conducted as an original and repeat study. In Cohort 6 (original and repeat), eight subjects were enrolled and administered the treatment. Six subjects received the active treatment and two received placebo. Active treatment of 150 μg treprostinil was administered via dry powder inhalation (DPI) as two capsules of 75 μg treprostinil strength, and placebo treatment was administered via DPI as two capsules of a placebo formulation. All inhalations were administered using the RS00 inhaler. Cohort 6 original included several mechanical device malfunctions and subjects who did not follow instructions, resulting in Cohort 6 repeat.

[0360]

[0471] Blood samples were taken at pre-dose and post-inhalation nominal concentrations of 0.083, 0.167, 0.25, Blood samples were collected at 0.33, 0.417, 0.5, 0.75, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, and 8 hours. Analysis of plasma concentration versus time data to calculate standard pharmacokinetic (PK) parameters after inhalation was performed using Phoenix WinNonlin version 6.3 with scheduled blood sampling times. Plasma concentrations were provided only for subjects on the active dose, and the number of randomized subjects was assigned by the bioanalytical laboratory to maintain study blinding.

[0361]

[0472] The table shown in Figure 8A is for the six active subjects in Cohort 6-R. Includes summary treprostinil concentration-time data for individual subjects along with descriptive statistics In two of the six subjects, peak concentrations occurred at 0.25 and 0.33 hours after inhalation. In the remaining two subjects, Tmax occurred at 0.167 and 0.417 hours after administration. At 4 hours after inhalation, four of the six active subjects had measurable concentrations of treprostinil. No subjects had quantifiable concentrations after the 6 or 8 hour time points.

[0362]

[0473] Pre-study non-compartmental PK of treprostinil for Cohort 6-R The parameters are summarized in the table shown in Figure 8B. Pre-study non-compartmental PK parameters of treprostinil for Cohort 6 - Original are summarized in the table shown in Figure 8C. Mean concentration-time data for each of the six cohorts are shown on a linear scale in Figure 8D. Cmax averaged 1.45 ng / mL, with the most frequent Tmax values observed at 0.25 and 0.33 hours post-inhalation. AUCinf values averaged 1.62 h*ng / mL, with a CV% of 68.3%. The apparent volume of distribution (Vz / F) averaged 107 L. Oral clearance (CL / F) averaged 126 L / h, ranging from 51.8 to 245. The variability (CV%) of CL / F values was 68.3%.

[0363]

[0474] In comparison, C in cohorts 1, 2, 3, 4, 5 and combined cohort 6 The Cmax averaged 0.364 ng / mL, 0.572 ng / mL, 0.728 ng / mL, 1.08 ng / mL, 1.19 ng / mL, and 1.21 ng / mL, respectively (Figure 8E), and the AUCinf values averaged 0.301 hr*ng / mL, 0.422 hr*ng / mL, 0.757 hr*ng / mL, 1.22 hr*ng / mL, 1.15 hr*ng / mL, and 1.37 hr*ng / mL (Figure 8F). Thus, a six-fold increase in dose from Cohort 1 to the combined Cohort 6 observations resulted in an approximately 260-400% increase in exposure, while a three-fold increase from Cohort 2 and a two-fold increase from Cohort 3 resulted in an approximately 130-255% and 81-98% increase in exposure, respectively. Furthermore, plots of the relationship between dose and Cmax and AUCinf are shown in Figure 8E and Figure 8F, respectively. These results indicate that both Cmax and AUCinf can increase proportionally with increasing dose. However, we observed some apparent device failures during the original 150 μg dose, which could result in incomplete and / or inefficient exposure. While no device failures were observed during the repeat dose, and the mean values may be higher than in the initial cohort, note the greater variability in the repeated cohort. A plot of the relationship between dose and CL / F (Figure 8G) shows that CL / F is independent of dose across the range of 25 to 150 μg treprostinil, suggesting that the PK of treprostinil is proportional to dose across the range of 25 to 150 μg treprostinil.

[0364]

[0475] Clinical Conclusions

[0476] LIQ861 is available as a single capsule (25, 50 and 75 mcg doses) or The combination of the two lower capsule strengths (100, 125, and 150 mcg doses) is administered at levels of 25, 50, 75, 100, 125, and 150 μg of treprostinil, with each capsule receiving either one or two breaths. According to an embodiment of the present invention, the active ingredient to particle powder ratios of the novel capsules, as well as breaths per capsule and powder per breath ratios for human administration, are included in the table below.

[0365] [Table 78]

[0366]

[0477] According to such an embodiment, each breath is 2.5 Particle potencies of ∼15 mg and 12.5 to 75 mcg of active agent may be received.

[0478] A predetermined amount of particulate powder filled into a capsule and delivered via a dry powder inhaler For a given treprostinil delivered, human clinical outcome results are included in the table below for LIQ861.

[0367] [Table 79]

[0368]

[0479] For comparison, TYVASO (United Therapeutics, In c.) provides the current standard of care for inhaled treprostinil therapy. This treprostinil is delivered via a nebulizer to treat PAH and is limited to delivering 6 mcg of treprostinil per breath, requiring nine breaths to reach a dose of 54 mcg. The current standard of care for inhaled therapy has been shown to be dose-limited to a maximum tolerated dose of 84 mcg of treprostinil, requiring 14 breaths to reach this dose. See Channick, R. et al., Inhaled Treprostinil: a Therapeutic Review, Drug Design, Development and Therapy 2012:6 19-28; and Nelsen AC et al., Pharmacokinetics of Inhaled Treprostinil Sodium in Healthy Volunteers. Am J Respir Crit Care Med. 2010; 181:A3348, both of which are incorporated herein by reference in their entirety.

[0369]

[0480] In another embodiment, the particles of the present invention comprise a loaded 0. It may contain 1% treprostinil loaded compared to 5% treprostinil. According to one embodiment of the present invention, the solid weight percentage in water is 1.06% treprostinil sodium, 92.44% trehalose dihydrate, 2% polysorbate 80, 4% L-leucine, 0.27% sodium citrate dihydrate, and 0.23% sodium chloride. A plurality of 1% treprostinil particles were prepared from a solution containing:

[0370]

[0481] According to the 1% treprostinil particle formulation of the present invention, the particle powder presented to the patient Mass and active ingredient include particle powder to active ingredient ratios for the novel capsules below, and powder ratios per breath per capsule and per breath for human administration.

[0371] [Table 80]

[0372]

[0482] According to such an embodiment, as shown in the table above, each breath is:1. You can receive particle potencies of 25-15mg and 12.5-150mcg of active agent.

[0373]

[0483] In the first clinical trial of LIQ861 involving delivery of a 150mcg dose For powder masses identified as acceptable, a 300 mcg active drug dose can be administered at 1% active drug particle size with a safe and acceptable powder mass and excipient amount.

[0374]

[0484] kit

[0485] According to an embodiment of the present invention, the dry powder inhaler device uses a capsule The LIQ861 particles can be combined into a kit containing the treprostinil powder. The capsules can be packaged in a blister pack with or without a desiccant, ensuring a controlled environment for the LIQ861 particle powder during transport with the user. The blister pack can contain single-dose capsules or multiple capsules for daily, weekly, or monthly administration. Typically, patients are treated four times daily for PAH indications. The kit can include capsules containing 25, 50, 75, 100, 125, 150, 200, 250, 300 mcg, or more dosage strengths for treating PAH. The powder particles in the capsules of the kit can be particles containing 0.5% treprostinil or 1% treprostinil.

[0375]

[0486]

[0376] [Table 81-1]

[0377] [Table 81-2]

[0378]

Table 81-3

[0379]

Table 81-4

[0380]

Table 81-5

[0381]

Table 81-6

[0382] Table 82

Claims

1. A dry powder inhalation treatment for pulmonary arterial hypertension comprising a dose of encapsulated dry particles containing more than 25 micrograms of treprostinil.

2. 10. The dry powder inhalation therapy of claim 1, wherein the dose of dry particles comprises 100 micrograms or more of treprostinil.

3. 10. The dry powder inhalation therapy of claim 1, wherein the dose of dry particles comprises 150 micrograms or more of treprostinil.

4. 10. The dry powder inhalation therapy of claim 1, wherein the dose of dry particles comprises 5 mg or more of dry particles.

5. 3. The dry powder inhalation therapy of claim 2, wherein the dose of dry particles comprises 10 mg or more of dry particles.

6. 4. The dry powder inhalation therapy of claim 3, wherein the dose of dry particles comprises 15 mg or more of dry particles.

7. A dry powder treatment for pulmonary arterial hypertension comprising a single capsule encapsulating 5 mg or more of dry particles, each 5 mg of dry particles containing 25 micrograms of treprostinil.

8. 1. A method of treating a patient suffering from pulmonary arterial hypertension, comprising: providing a dry powder inhaler to a patient; Providing a patient with at least one capsule for use in a dry powder inhaler, the capsule containing at least 25 micrograms of treprostinil; instructing the patient to inhale treprostinil using a dry powder inhaler; A method comprising:

9. 9. The method of claim 8, wherein the capsule contains at least 50 micrograms of treprostinil.

10. 9. The method of claim 8, wherein the capsule contains at least 100 micrograms of treprostinil.

11. 9. The method of claim 8, wherein the capsule contains at least 150 micrograms of treprostinil.

12. 1. A method of treating a patient suffering from pulmonary arterial hypertension, comprising administering a dry powder dose of treprostinil to a patient suffering from pulmonary arterial hypertension, wherein the dose of treprostinil is greater than 85 micrograms.

13. A dry powder inhalation composition for treating pulmonary arterial hypertension, comprising a plurality of dry powder particles comprising treprostinil, a non-reducing sugar, a humectant, a hydrophobicity adjuster, a pH adjuster, and a buffering agent.

14. 14. A dry powder inhalation composition according to claim 13, wherein the bulking agent comprises trehalose dihydrate.

15. 14. A dry powder inhalation composition according to claim 13, wherein the humectant comprises polysorbate 80.

16. 14. The dry powder inhalation composition of claim 13, wherein the hydrophobicity modifier comprises L-leucine.

17. 14. A dry powder inhalation composition according to claim 13, wherein the pH adjuster comprises sodium citrate dihydrate.

18. 14. A dry powder inhalation composition according to claim 13, wherein the buffer comprises sodium chloride.

19. 14. The dry powder inhalation composition of claim 13, wherein the composition comprises less than about 4 percent by weight water.

20. 14. The dry powder inhalation composition of claim 13, wherein the composition comprises less than about 2 percent by weight water.

21. 14. The dry powder inhalation composition of claim 13, wherein the composition comprises less than about 1 weight percent water.

22. 14. The dry powder inhalation composition of claim 13, wherein the dry powder particles comprise particles having a three-dimensional shape comprising a width and length of between 1 micrometer and 2 micrometers, and a depth of between 0.3 micrometer and 0.8 micrometers.

23. 14. The dry powder inhalation composition of claim 13, wherein the dry powder particles comprise a drying solution comprising trehalose dihydrate, L-leucine, treprostinil sodium, polysorbate 80, sodium citrate dihydrate, sodium chloride, and water.

24. 24. The dry powder inhalation composition of claim 23, wherein the dry powder particles comprise, by solids percentage, about 0.581 percent treprostinil sodium, about 92.32 percent trehalose, about 2.19 percent polysorbate 80, about 4.39 percent L-leucine, about 0.26 percent sodium citrate, and about 0.25 percent sodium chloride.

25. 1. A method of producing particles for dry powder delivery to the lungs of a patient in need thereof, comprising: forming a composition comprising about 12.30 weight percent trehalose dihydrate, about 0.53 weight percent L-leucine, about 0.07 weight percent treprostinil sodium, about 0.26 weight percent polysorbate 80, about 0.04 weight percent sodium citrate dihydrate, about 0.03 weight percent sodium chloride, and about 86.78 weight percent water into particles; drying the composition so that the particles contain less than 4 weight percent water; A method comprising:

26. 1. A method of treating a patient suffering from pulmonary arterial hypertension, comprising delivering greater than 12.5 micrograms per breath of treprostinil to the patient.

27. 1. A method of treating a patient suffering from pulmonary arterial hypertension, comprising delivering greater than 25 micrograms per breath of treprostinil to the patient.

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