Methods for treating cancer with inhaled angiogenesis inhibitors

Abstract

The present disclosure relates to methods of treating cancer, e.g., lung cancer (e.g., non-small cell lung cancer), by administering a dry powder to the respiratory tract of a subject in need thereof (e.g., by oral inhalation), the dry powder comprising respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) and, optionally, a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., leucine and a sodium salt, e.g., sodium sulfate).

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Application No. 63 / 399,080, filed August 18, 2022, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Lung cancer is the third most common type of cancer and the leading cause of cancer death in the United States. The five-year survival rate for lung cancer is only 56 percent when it is detected at a localized stage, and only 16 percent of lung cancer cases are diagnosed at this early stage. If lung cancer spreads to other organs, the five-year survival rate drops to just 5%. Thus, more than half of lung cancer patients die within one year of diagnosis. (See, e.g., US National Institute of Health, National Cancer Institute. SEER Cancer Statistics Review, 1975-2015.) Therefore, new and effective lung cancer treatments are urgently needed.

[0003] Because non-small cell lung cancer (NSCLC) is a highly vascularized tumor, angiogenesis inhibitors can be used alone or in addition to other cancer treatments (e.g., neoadjuvant treatment) to treat NSCLC patients. However, there are only three FDA-approved angiogenesis inhibitors for NSCLC, and these drugs can cause serious side effects and mortality, and they offer only modest benefits in terms of overall survival and are expensive. (See, e.g., Aftab et al. Cancer Res. (2011) 71:6764-6772; Daum et al. Front. Cell Dev. Biol. (2021) 9:1-17.)

[0004] Itraconazole has recently been identified as an antiangiogenic agent and an antagonist of the Hedgehog signaling pathway, and is being investigated for the treatment of NSCLC. Specifically, when high-dose oral itraconazole was administered to clinical trial subjects with NSCLC, itraconazole tissue concentrations were found to be significantly associated with reduced tumor volume and perfusion, decreased angiogenesis-stimulating cytokines IL1b and GM-CSF, and decreased tumor microvascular density (Gerber et al. Clin. Cancer Res. (2020) 26:6017-6027).

[0005] Itraconazole is a well-known small molecule drug that has been available for over 30 years. It is typically used to treat fungal infections and is the active ingredient in the antifungal drug SPORANOX® (itraconazole, Janssen Pharmaceuticals). Itraconazole can be synthesized using a variety of methods well known in the art. Therefore, itraconazole appears to be a suitable alternative to the angiogenesis inhibitors currently approved for the treatment of NSCLC. However, itraconazole has poor water solubility, poor oral bioavailability, unpredictable and heterogeneous pharmacokinetic parameters, and extensive drug-drug interactions. As a result, itraconazole generally causes side effects and toxicity in patients, and pharmaceutical formulations that deliver safe therapeutic levels of itraconazole have been difficult to obtain. For example, in clinical trials of high-dose oral itraconazole in NSCLC patients, pharmacokinetic parameters varied more than six-fold across the patient population, and only two subjects achieved the predicted plasma concentrations of itraconazole (Gerber, supra). Considering the significant association between itraconazole tissue concentrations and reduced tumor volume, the unpredictable pharmacokinetics of oral itraconazole is a significant obstacle to its application in cancer therapy, and its poor pharmacokinetics and significant side effects further limit its use in therapy.

[0006] There is a need for new methods of treating cancer, particularly lung cancer, using formulations of angiogenesis inhibitors that can consistently achieve therapeutically effective concentrations in cancer tissues while avoiding high plasma concentrations and toxicity. Summary of the Invention

[0007] The present disclosure relates to methods of treating cancer, e.g., lung cancer, e.g., non-small cell lung cancer (NSCLC), by administering to a subject in need thereof by inhalation (e.g., oral inhalation) a respirable dry powder comprising homogenous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) and, optionally, a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., sodium salts and leucine).

[0008] The cancer treated by the methods disclosed herein may be lung cancer, for example, NSCLC. The cancer (e.g., lung cancer) may be locally advanced cancer. In some embodiments, the cancer (e.g., lung cancer) is metastatic cancer (or stage IV cancer). The cancer (e.g., lung cancer) may be refractory, recurrent, or both.

[0009] The methods disclosed herein may include administering the dry powder to a subject with an additional therapeutic agent. The additional therapeutic agent may be a therapeutic agent disclosed herein, such as a chemotherapeutic agent, a targeted cancer therapy, an immunotherapy, or a combination thereof. In some embodiments, the additional therapeutic agent is a chemotherapeutic agent. In some embodiments, the additional therapeutic agent is an immunotherapy agent. In some embodiments, the additional therapeutic agent is a targeted cancer therapy.

[0010] In some aspects, methods of treating cancer (e.g., lung cancer) disclosed herein include (i) administering to the respiratory tract of a subject in need thereof a dry powder, the dry powder comprising homogenous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) and, optionally, a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., leucine and sodium salts); and (ii) administering to the subject an additional therapeutic agent, e.g., a chemotherapeutic agent, a targeted cancer therapy agent, an immunotherapy agent, or a combination thereof.

[0011] In some embodiments, methods of treating cancer (e.g., lung cancer) disclosed herein include (i) administering to the respiratory tract of a subject in need thereof a dry powder, the dry powder comprising homogeneous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) and, optionally, a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., leucine and sodium salts); (ii) orally administering itraconazole to the subject; and (iii) optionally administering to the subject an additional therapeutic agent (e.g., a chemotherapeutic agent, a targeted cancer therapy agent, an immunotherapy agent, or a combination thereof).

[0012] In the methods disclosed herein that include administering an additional therapeutic agent and / or orally administering itraconazole in addition to the dry powder of the present invention, the dry powder may be administered at any time and in any order relative to the additional therapeutic agent and / or oral itraconazole. For example, the additional therapeutic agent and / or oral itraconazole may be administered at a dose within 24 hours of administering the dry powder at a dose before or after administering the dry powder at a dose, e.g., within about 12 hours, about 6 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, about 30 minutes, or less, of the dry powder. Alternatively, the dry powder may be administered to the subject more than one day before or after, e.g., two, three, four, five, six, one, two, two, three, four, or more days before or after, the administration of the additional chemotherapeutic agent and / or oral itraconazole. In some embodiments, the dry powder is administered to the subject about 1 to about 28 days before the administration of the additional chemotherapeutic agent and / or oral itraconazole, e.g., 1 to about 21 days, about 1 to about 18 days, about 1 to about 16 days, about 1 to about 14 days, about 1 to about 12 days, or about 1 to about 10 days before the administration of the additional chemotherapeutic agent and / or oral itraconazole.

[0013] In some embodiments, the additional therapeutic agent, e.g., a platinum-based agent, antimetabolite, anti-microtubule agent, topoisomerase inhibitor, an anthracycline, a KRAS (Kirsten rat sarcoma viral oncogene homolog) inhibitor, an ALK (anaplastic lymphoma kinase) inhibitor, an EGFR (epidermal growth factor receptor) inhibitor, a VEGF (vascular endothelial growth factor) inhibitor, a BRAF inhibitor, a MEK inhibitor, a RET inhibitor, a MET inhibitor, or an immunotherapy (e.g., an immune checkpoint inhibitor, e.g., a PD-1 / PD-L1 inhibitor or a CTLA-4 inhibitor), is used to treat cancer. In some embodiments, the additional therapeutic agent is pemetrexed, cisplatin, carboplatin, oxaliplatin, satraplatin, picoplatin, nedaplatin, triplatin, lipoplatin, paclitaxel, docetaxel, doxorubicin, gemcitabine, vinorelbine, etoposide, SN-38, camptothecin, topotecan, exatecan, irinotecan, belotecan, methotrexate, bevacizumab, ranibizumab, aflibercept, ramucirumab, nintedanib, erlotinib, afatinib, axitinib, gefitinib, cabozantinib, osimertinib, dacomitinib, sotrasib, crizotinib, ribozyme, ribozyme inhibitors (RIRIs), ... and combinations thereof.

[0014] In some embodiments, the dry powders of the present invention are administered to a subject before, concurrently with, or after surgery, e.g., surgery to remove cancerous tissue from the subject (e.g., segmentectomy, sleeve resection, wedge resection, lobectomy, pneumonectomy, lymphadenectomy, or a combination thereof).

[0015] In some embodiments, the dry powder is administered to the subject prior to, concurrently with, or after radiation therapy to treat cancer (e.g., brachytherapy, external beam radiation therapy (EBRT), stereotactic body radiation therapy (SBRT), stereotactic ablative body radiation therapy (SABR), three-dimensional conformal radiation therapy (3D-CRT), intensity-modulated radiation therapy (IMRT), intensity-modulated arc therapy (VMAT), stereotactic radiosurgery (SRS), radiofrequency ablation, or a combination thereof).

[0016] In some embodiments, the dry powder angiogenesis inhibitor administered to a subject has a crystallinity of at least 50%, for example, 60% crystallinity, 70% crystallinity, 80% crystallinity, 90% crystallinity, 92% crystallinity, 94% crystallinity, 95% crystallinity, 96% crystallinity, 97% crystallinity, 98% crystallinity, 99% crystallinity, 99.5% crystallinity, or 99.9% crystallinity or greater. In some embodiments, the angiogenesis inhibitor is itraconazole. The itraconazole may be crystalline itraconazole. In some embodiments, the itraconazole is not amorphous itraconazole.

[0017] The dry powders used in the methods disclosed herein may contain the angiogenesis inhibitor in the form of primary particles, the primary particles being about 50 nm to about 5,000 nm (Dv50), e.g., about 50 nm to about 800 nm (Dv50), about 50 nm to about 300 nm (Dv50), about 50 nm to about 200 nm (Dv50), or about 100 nm to about 300 nm (Dv50). In some embodiments, the angiogenesis inhibitor is a crystalline primary particle.

[0018] The angiogenesis inhibitor may be present in the respirable dry particles in an amount of about 1% to about 95% by weight, e.g., about 40% to about 90% by weight, about 55% to about 85% by weight, about 55% to about 75% by weight, about 65% to about 85% by weight, or about 40% to about 60% by weight.

[0019] In some embodiments, the dry powder comprises a stabilizer (e.g., polysorbate 80) and one or more excipients (e.g., sodium sulfate and leucine). The ratio of angiogenesis inhibitor to stabilizer (wt:wt) in the respirable dry particles can be any desired ratio, for example, about 1:1 to 50:1, 10:1 or more, about 10:1, about 20:1, about 5:1 to about 20:1, about 7:1 to about 15:1, or about 9:1 to about 11:1. In some embodiments, the stabilizer is present in the respirable dry particles in an amount of about 0.05% to about 45% by weight, e.g., about 4% to about 10% by weight. The one or more excipients can be present in the respirable dry particles in an amount of about 10% to about 99% by weight. For example, the one or more excipients can be present in the respirable dry particles in an amount of about 5% to about 50% by weight.

[0020] The one or more excipients present in the dry powder may include a monovalent metal cation salt, a divalent metal cation salt, an amino acid, a sugar alcohol, or a combination thereof. In some embodiments, the one or more excipients include a sodium salt and an amino acid. The sodium salt may be selected from the group consisting of sodium chloride and sodium sulfate. The amino acid may be leucine.

[0021] In some embodiments, the stabilizer present in the respirable dry particles can be polysorbate 80 in an amount of 10 wt% or less, e.g., 7 wt% or less, 5 wt% or less, 4 wt% or less, 3 wt% or less, 2 wt% or less, or 1 wt% or less.

[0022] In some embodiments, the stabilizer present in the respirable dry particles is oleic acid or a salt thereof in an amount of 10 wt% or less, e.g., 7 wt% or less, 5 wt% or less, 4 wt% or less, 3 wt% or less, 2 wt% or less, or 1 wt% or less.

[0023] In some embodiments, the respirable dry particles have (i) a volume median geometric diameter (VMGD) of about 10 micrometers or less, e.g., about 5 micrometers or less, (ii) a tap density of about 0.2 g / cc or more, e.g., a tap density of 0.2 g / cc to 1.0 g / cc, (iii) a dispersibility ratio at 1 bar / 4 bar (1 / 4 bar) of less than about 1.5, as measured by laser diffraction, and / or (iv) a dispersibility ratio at 0.5 bar / 4 bar (0.5 / 4 bar) of about 1.5 or less, as measured by laser diffraction.

[0024] In some embodiments, the dry powder has (i) a mass median aerodynamic diameter (MMAD) of about 1 micrometer to about 5 micrometers, and / or (ii) a fine particle fraction (FPF) of less than 5 micrometers of the total dose of about 25% or more.

[0025] In some embodiments, the respirable dry particles, when emitted from a passive dry powder inhaler having a resistance of about 0.036 sqrt (kPa) / liter per minute using a No. 3 capsule containing a total mass of 10 mg (said total mass consisting of the respirable dry particles) at an inhalation flow rate of 30 LPM over a 3 second period, have a capsule emitted powder mass of at least 80%, and the respirable dry particles emitted from the inhaler have a volume median geometric diameter of 5 micrometers or less as measured by laser diffraction.

[0026] The methods disclosed herein may include administering the dry powder to the respiratory tract of a subject using a passive dry powder inhaler, for example, a capsule-based passive dry powder inhaler.

[0027] In some embodiments, the dry powder provides a sputum lung concentration (e.g., steady-state concentration in sputum) of the angiogenesis inhibitor (e.g., itraconazole) of at least 100 ng / mL, e.g., about 500 ng / mL, about 800 ng / mL, about 1200 ng / mL, about 1600 ng / mL, about 2000 ng / mL, about 3000 ng / mL, about 4000 ng / mL, about 5000 ng / mL, about 6000 ng / mL, about 7000 ng / mL, about 8000 ng / mL, about 10,000 ng / mL, about 15,000 ng / mL, about 20,000 ng / mL, about 25,000 ng / mL, about 50,000 ng / mL, about 75,000 ng / mL, about 100,000 ng / mL, ng / mL, about 125,000ng / mL, about 150,000ng / mL, about 175,000ng / mL, or about 200,000ng / mL or more, or about 500ng / mL to 400,000ng / mL, about 500ng / mL to 300,000ng / mL, about 500ng / mL to 200,000ng / mL, about 500ng / mL to 100,000ng / mL, about 500ng / mL to 50,000ng / mL, about 500ng / mL to 25,000ng / mL, about 500ng / mL to 10,000ng / mL, about 2000ng / mL to 8000ng / mL, or about 2000ng / mL to 8100ng / mL.

[0028] In some embodiments, the dry powder provides a lung tissue concentration (e.g., steady-state concentration in the tissue) of the angiogenesis inhibitor (e.g., itraconazole), e.g., a concentration in lung tumor tissue, of at least about 100 ng / g, e.g., about 500 ng / g, about 800 ng / g, about 1200 ng / g, about 1600 ng / g, about 2000 ng / g, about 3000 ng / g, about 4000 ng / g, about 5000 ng / g, about 6000 ng / g, about 7000 ng / g, about 8000 ng / g, about 9000 ng / g, or about 10,000 ng / g or more, e.g., up to about 1 0.6 mg / g, at most about 1.4 mg / g, at most about 1.2 mg / g, or at most about 1.0 mg / g, or about 100 ng / g to about 900,000 ng / g, for example, about 100 ng / g to about 800,000 ng / g, about 100 ng / g to about 700,000 ng / g, about 100 ng / g to about 600,000 ng / g, about 100 ng / g to about 500,000 ng / g, about 100 ng / g to about 400,000 ng / g, about 100 ng / g to about 300,000 ng / g, about 100 ng / g to about 200,000 ng / g, or about 100 ng / g to about 1 00,000ng / g, or about 100ng / g to about 1.6mg / g, for example, about 500ng / g, about 1000ng / g, about 1500ng / g, about 2000ng / g, about 2500ng / g, about 5000ng / g, about 10,000ng / g, about 15,000ng / g, about 20,000ng / g, about 25,000ng / g, about 30,000ng / g, about 40,000ng / g, about 50,000ng / g, about 60,000ng / g, about 70,000ng / g, about 80,000ng / g, about 90,000ng / g, about 100,000ng / g, about 12 The antibody is administered in an amount effective to achieve a saturation of about 0,000 ng / g, about 140,000 ng / g, about 160,000 ng / g, about 180,000 ng / g, about 200,000 ng / g, about 250,000 ng / g, about 300,000 ng / g, about 350,000 ng / g, about 400,000 ng / g, about 450,000 ng / g, about 500,000 ng / g, about 600,000 ng / g, about 700,000 ng / g, about 800,000 ng / g, about 900,000 ng / g, about 1 mg / g, about 1.2 mg / g, about 1.4 mg / g, or about 1.6 mg / g.

[0029] In some embodiments, the dry powder is administered in an amount effective to achieve a plasma concentration (eg, steady state concentration) of the angiogenesis inhibitor (eg, itraconazole) of 25 ng / mL or less.

[0030] In some embodiments, the dry powder is administered to a subject as a single dose (e.g., a single dose administered via oral inhalation). In some embodiments, the dry powder is administered as an initial dose followed by one or more subsequent doses.

[0031] In another aspect, the disclosure relates to a dry powder (e.g., a dry powder disclosed herein) for use in treating cancer (e.g., lung cancer, e.g., NSCLC), the dry powder comprising homogenous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) and, optionally, a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., sodium salts and leucine), the dry powder for administration to the respiratory tract of a subject.

[0032] In another aspect, the disclosure relates to the use of a dry powder (e.g., a dry powder disclosed herein) in the manufacture of a medicament for the treatment of cancer (e.g., lung cancer, e.g., NSCLC), the dry powder comprising homogenous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) and, optionally, a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., sodium salts and leucine), the medicament for administration to the respiratory tract of a subject. [Brief explanation of the drawings]

[0033] [Figure 1] 1 is an X-ray diffraction plot of particles of Formulations I and II. [Figure 2] 1 is an X-ray diffraction plot of particles of Formulations III and IV. [Figure 3] 10 is an X-ray diffraction plot of particles of Formulations V and VI. [Figure 4] 10 is an X-ray diffraction plot of particles of Formulations VII and VIII. [Figure 5]1 is an X-ray diffraction plot of particles of Formulation XI. [Figure 6] 10 is an X-ray diffraction plot of particles of Formulation XII. [Figure 7] 10 is an X-ray diffraction plot of particles of Formulation XIII. [Figure 8] 10 is an X-ray diffraction plot of particles of Formulation XIV. [Figure 9] 10 is an X-ray diffraction plot of particles of Formulation XV. [Figure 10] 10 is an X-ray diffraction plot of particles of Formulation XVI. [Figure 11] 10 is an X-ray diffraction plot of particles of Formulation XIX. [Figure 12] 1 is a plot showing the cumulative mass dissolution of impactor stage mass (ISM) collected in a paddle-over-disk (POD) dissolution test in a USP Apparatus II setup after aerosolization of various dry powders containing the angiogenesis inhibitor itraconazole from a UniDose RS01 dry powder inhaler (DPI) at 60 L / min. [Figure 13] 1 is a plot showing the cumulative mass dissolution (%) of ISM collected in a POD dissolution test in a USP Apparatus II setup after aerosolization of various dry powders containing the angiogenesis inhibitor itraconazole from RS01 DPI at 60 L / min with a UniDose. [Figure 14A] 1 is a plot showing the relationship between dissolution half-life and particle size of the angiogenesis inhibitor itraconazole in various dry powders. [Figure 14B] 1 is a plot showing the dissolution half-life versus surface area of ​​the angiogenesis inhibitor itraconazole in various dry powders. [Figure 15] 1 is a plot showing the relationship between dissolution half-life and Cmax of the angiogenesis inhibitor itraconazole in various dry powders. [Figure 16] 1 is a plot showing the relationship between dissolution half-life and dose-adjusted Cmax (expressed as a ratio to Formulation XIX) for various dry powders containing the angiogenesis inhibitor itraconazole. [Figure 17]1 is a plot showing the cumulative mass dissolution (%) of ISM collected in a POD dissolution test in a USP Apparatus II setup after aerosolization of an itraconazole suspension formulation from a UniDose Micro Mist nebulizer at 15 L / min. [Figure 18] 1 is a plot showing the cumulative percent mass recovery from various dry powders, including the angiogenesis inhibitor itraconazole, deposited on stage 4 of a cNGI. [Figure 19] 1 is a plot showing the relationship between dissolution half-life and Cmax of the angiogenesis inhibitor itraconazole in various dry powders. [Figure 20] 1 is a plot showing the relationship between diffusion rate and dose-adjusted Cmax (expressed as a percentage of Formulation XIX) in various dry powders containing the angiogenesis inhibitor itraconazole. [Figure 21] 1 is a plot showing the cumulative percent mass recovered from a nebulized suspension formulation of the angiogenesis inhibitor itraconazole deposited on stage 4 of a cNGI. [Figure 22] Figures 1A and 1B show simulated kinetics of Formulations XIX, XII, and SPORANOX® in terms of plasma exposure (A) and lung exposure (B) using a model established from SPORANOX® animal pharmacokinetic (PK) data and human data. In both simulations, 5 mg was inhaled once daily (Formulations XIX and XII), while a 200 mg SPORANOX® oral solution dose was administered twice daily. Concentrations of the angiogenesis inhibitor itraconazole were measured over 7 days of administration. [Figure 23]Figures 1A and 1B show simulated kinetics of Formulations XIX, XII, and SPORANOX® in terms of plasma exposure (A) and lung exposure (B) using a model established from SPORANOX® PK data and human data. In each case, 20 mg was inhaled once daily (Formulations XII and XIX), while a 200 mg SPORANOX® oral solution dose was administered twice daily. Concentrations of the angiogenesis inhibitor itraconazole were measured over 7 days of administration. [Figure 24] 1 is a graph showing the plasma pharmacokinetic profile over 96 hours following a single dose of Formulation XII in healthy volunteers, details of which study are provided in Example 21. [Figure 25] 1 is a graph showing the 24-hour plasma pharmacokinetic profile of Formulation XII after a single dose or 14 daily doses in healthy volunteers, details of which study are provided in Example 21. [Figure 26] Figures A and B show summary data of systemic pharmacokinetics in asthmatic patients after a single inhaled or oral dose. Pharmacokinetic profile of the angiogenesis inhibitor itraconazole in sputum (A) and plasma (B) following a single dose of Formulation XII (▲) or oral SPORANOX® (△) in asthmatic patients. DETAILED DESCRIPTION OF THE INVENTION

[0034] The present disclosure relates to a method of treating cancer (e.g., lung cancer, e.g., NSCLC) by administering a respirable dry powder by inhalation (e.g., oral inhalation) to a subject in need thereof, wherein the dry powder comprises homogenous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole). In a preferred embodiment, the angiogenesis inhibitor is itraconazole (e.g., crystalline itraconazole). The dry powder may further comprise a stabilizer. In a preferred embodiment, the stabilizer is polysorbate 80 (PS80). The dry powder may further comprise one or more excipients. In a preferred embodiment, the one or more excipients are leucine and a sodium salt (e.g., sodium sulfate).

[0035] The inventors have discovered that administration of the dry powders disclosed herein can result in lung concentrations of angiogenesis inhibitors (e.g., itraconazole) that are substantially higher than those achieved by oral administration. Without wishing to be bound by theory, it is believed that such administration is effective in treating cancer (e.g., lung cancer) by increasing the lung concentration of angiogenesis inhibitors using the dry powders disclosed herein, while minimizing the systemic concentration of the angiogenesis inhibitor, thereby preventing side effects and toxicity. It is further believed that therapeutic concentrations of angiogenesis inhibitors in the lungs can be achieved at total doses that are relatively low compared to conventional administration methods. For example, studies involving the treatment of NSCLC with oral itraconazole required a dose of 600 mg / day, which is significantly higher than the standard recommended dose of itraconazole (approximately 200 mg / day).

[0036] Administration of the dry powders disclosed herein can result in a relatively high lung:systemic concentration ratio of an angiogenesis inhibitor (e.g., itraconazole). Without wishing to be bound by any particular theory, it is believed that a relatively high lung:systemic concentration ratio can minimize off-target effects and / or toxicity while achieving effective cancer treatment. Furthermore, by achieving a high drug ratio in the lungs, a relatively low dose of the angiogenesis inhibitor is administered to a subject, compared to the large doses required for oral or intravenous administration. Additionally, the inventors have discovered that administration of the dry powders disclosed herein can result in a more consistent exposure of the angiogenesis inhibitor (e.g., across multiple doses or within a subject population) relative to other formulations or administration routes of the angiogenesis inhibitor (e.g., compared to oral doses of itraconazole). Thus, the methods disclosed herein offer advantages over current applications of angiogenesis inhibitors, which typically involve oral administration of large amounts and fail to achieve predictable or consistent concentrations in cancerous tissue (e.g., lung tissue).

[0037] The dry powder disclosed herein can be administered to a subject by inhalation, for example, oral inhalation. To achieve oral inhalation, a dry powder inhaler, for example, a passive dry powder inhaler, can be used. The dry powder disclosed herein can be used to treat cancer, for example, lung cancer (e.g., NSCLC), in a subject. The relatively low-dose inhaled formulation of angiogenesis inhibitors (e.g., itraconazole) minimizes many of the drawbacks of oral or intravenous (IV) formulations in the treatment of these patients.

[0038] The respirable dry powders disclosed herein comprise an angiogenesis inhibitor. The angiogenesis inhibitor may be in crystalline microparticle form. In a preferred embodiment, the angiogenesis inhibitor is itraconazole in crystalline microparticle form. Respirable dry powders comprising itraconazole for use in treating fungal infections are described in WO2018 / 071757, WO2019 / 204583, and WO2019 / 204597, the entire contents of which are incorporated herein by reference.

[0039] The inventors have discovered that dry powders comprising itraconazole in amorphous form, when inhaled at therapeutic doses, have a shorter lung residence time, resulting in a lower lung to plasma exposure ratio and a lower undesirable toxic effect on lung tissue. Without wishing to be bound by any particular theory, it is believed that the dry powders disclosed herein that comprise an angiogenesis inhibitor (e.g., itraconazole) in a crystalline form (e.g., nanocrystalline form) have a slower dissolution rate in the lung relative to the amorphous form, resulting in a more continuous exposure over a 24-hour period following administration and minimizing systemic exposure.

[0040] Additionally, the local toxicity observed in lung tissue without amorphous administration is not related to the total exposure of the lung tissue to itraconazole, in terms of total dose or duration of exposure. For example, itraconazole has no known activity against human or animal lung cells, so elevated local concentrations lack local pharmacological activity to explain its local toxicity. Instead, the toxicity of the amorphous form appears to be related to the increased solubility of itraconazole due to its amorphous nature, resulting in supersaturation of the interstitial space with itraconazole and subsequent recrystallization within the tissue, leading to local granulomatous inflammation. Surprisingly, the inventors have discovered that the dry powders disclosed herein containing itraconazole in crystalline particulate form are less toxic to lung tissue than more rapidly dissolving formulations, such as those containing amorphous itraconazole. This was surprising. Crystalline microparticulate itraconazole formulations have lower aqueous solubility than formulations containing amorphous itraconazole, resulting in a slower dissolution rate and longer lung retention than a comparable dose of amorphous itraconazole. Furthermore, crystalline microparticulate itraconazole results in higher lung exposures than a comparable dose of amorphous itraconazole after a single dose and over 28 days.

[0041] The degree of crystallinity of the angiogenesis inhibitor, the size of the crystalline primary particles of the angiogenesis inhibitor, and the identity and amount of excipients and stabilizers all appear to be important factors for effective therapy and reduced toxicity in the lungs. Without wishing to be bound by any particular theory, it is believed that angiogenesis inhibitors with smaller crystalline particles (e.g., nanocrystalline or microcrystalline angiogenesis inhibitors) dissolve more rapidly in airway lining fluid than larger crystalline particles, in part due to their greater total surface area. It is also believed that crystalline angiogenesis inhibitors dissolve more slowly in airway lining fluid than amorphous angiogenesis inhibitors, in part due to their reduced water solubility. Thus, the dry powders described herein can be formulated with a crystalline particulate form of an angiogenesis inhibitor (e.g., itraconazole) that provides a desired crystal size or crystal size range of the angiogenesis inhibitor within the dry powder, optionally with suitable excipients and stabilizers in appropriate ratios with the angiogenesis inhibitor, each of which can be tailored to, for example, affect dissolution rate and provide desired pharmacokinetic properties while avoiding unacceptable toxicity in the lung.

[0042] Respirable dry powders for use in the methods disclosed herein may comprise homogeneous respirable dry particles comprising 1) an angiogenesis inhibitor (e.g., itraconazole, e.g., itraconazole in crystalline microparticle form), 2) a stabilizer, and, optionally, 3) one or more excipients. Such respirable dry particles can be prepared using any suitable method, for example, by preparing a feedstock in which the angiogenesis inhibitor (e.g., itraconazole, e.g., itraconazole in crystalline microparticle form) is suspended in an aqueous solution of an excipient and spray-drying the feedstock.

[0043] The dry powder may be administered to a subject by inhalation, for example, oral inhalation. Oral inhalation can be achieved using a dry powder inhaler, such as a passive dry powder inhaler. The dry powder can be used to treat cancer, such as lung cancer, or more specifically, non-small cell lung cancer (NSCLC), in a subject. The inhaled formulations of angiogenesis inhibitors (e.g., itraconazole) disclosed herein minimize many of the drawbacks of oral or intravenous (IV) formulations in treating these subjects. For example, oral or intravenous formulations of angiogenesis inhibitors may not consistently achieve therapeutic concentrations at the site of cancer cells or tumors (e.g., in the lungs) or may require relatively high doses to achieve therapeutic lung concentrations, resulting in toxicity or side effects. The high doses required for oral administration also increase the risk of drug-drug interactions (DDIs). For example, administering an oral angiogenesis inhibitor (e.g., itraconazole) in combination with a standard of care protocol for cancer treatment may be contraindicated due to DDIs, e.g., when the standard of care involves administering a cancer therapeutic agent that is a substrate for the same enzyme as the angiogenesis inhibitor, e.g., a cytochrome P450, e.g., CYP3A4. Thus, the high dose required for oral administration of the angiogenesis inhibitor may limit its use in combination with other cancer therapeutic agents or as neoadjuvant therapy. The dry powders disclosed herein can reliably achieve therapeutic concentrations of the angiogenesis inhibitor (e.g., itraconazole) in a subject (e.g., in lung tissue), thereby reducing the total dose of the angiogenesis inhibitor required for administration to a subject and also allowing for relatively low systemic concentrations (e.g., plasma concentrations) of the angiogenesis inhibitor, thereby achieving efficacy (e.g., in treating cancer, e.g., lung cancer) while reducing side effects and toxicity.Advantageously, the relatively small amounts of angiogenesis inhibitor required when using the dry powders disclosed herein can reduce the risk of DDIs compared to the larger amounts required for oral administration, thereby providing the opportunity to combine the dry powder containing angiogenesis inhibitor with another agent, e.g., a standard of care treatment for NSCLC, e.g., using the dry powders disclosed herein as neoadjuvant therapy with a reduced risk of DDIs.

[0044] The inhaled formulations of angiogenesis inhibitors (e.g., itraconazole) disclosed herein may provide synergistic effects when combined with another chemotherapeutic agent. Accordingly, the present disclosure outlines methods that include combination therapy. For example, administering the respirable dry powder disclosed herein to a subject in addition to another therapy (e.g., an additional therapeutic agent and / or radiation) may improve the extent of tumor volume reduction in the subject compared to the extent of tumor volume reduction achieved when the same treatment is administered without the dry powder.

[0045] definition As used herein, the term "about" refers to a relative range of ±5% of the stated value, for example, "about 20 mg" would be "20 mg ±1 mg."

[0046] As used herein, the term "administration" or "administering" refers to introducing a composition comprising a therapeutic agent into a subject. For example, administering may refer to introducing respirable dry particles disclosed herein into the respiratory tract of a subject.

[0047] As used herein, the term "amorphous" refers to the absence of significant crystallinity when analyzed via powder X-ray diffraction (XRD).

[0048] The term "capsule emitted powder mass" or "CEPM," as used herein, refers to the amount of dry powder emitted from a capsule or dose unit container upon actuation of a dry powder inhaler, e.g., during inhalation. CEPM is typically measured gravimetrically by weighing the capsule before and after emission to determine the emitted powder mass. CEPM can be expressed as the emitted powder mass in milligrams or as a percentage of the original powder mass filled in the capsule before emission.

[0049] The term "crystalline particulate form," as used herein, refers to an angiogenesis inhibitor (e.g., itraconazole) (including pharmaceutically acceptable forms thereof, including salts, hydrates, enantiomers, etc.) that is in the form of particles (i.e., primary particles smaller than the respirable dry particles that comprise the dry powders disclosed herein), in which the angiogenesis inhibitor is at least about 50% crystalline. The percent crystallinity of an angiogenesis inhibitor refers to the proportion of compound that is in a crystalline state relative to the total amount of compound present in the primary particles. If desired, the angiogenesis inhibitor can be at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or about 100% crystalline. Angiogenesis inhibitors in crystalline microparticle form are in the form of particles having a median volume diameter (Dv50) of about 50 nanometers (nm) to about 5,000 nm, preferably a Dv50 of 80 nm to 1750 nm, or preferably a Dv50 of 50 nm to 800 nm.

[0050] The term "dispersibility" is a term of art that describes a characteristic of a dry powder or respirable dry particle that is dispersed into a respirable aerosol. Dispersibility of a dry powder or respirable dry particle is expressed herein, in one embodiment, as the volume median geometric diameter (VMGD) measured at 1 bar dispersion pressure (i.e., regulated pressure) divided by the VMGD measured at 4 bar dispersion pressure (i.e., regulated pressure), or the VMGD at 0.5 bar divided by the VMGD at 4 bar, as measured by laser diffraction, e.g., laser diffraction on a HELOS / RODOS. These quotients are referred to herein as the "1 bar / 4 bar dispersibility ratio" and "0.5 bar / 4 bar dispersibility ratio," respectively, with higher dispersibility correlated with lower quotients. For example, the 1 bar / 4 bar dispersibility ratio refers to the VMGD of a dry powder or respirable dry particle emitted from the orifice of a RODOS dry powder disperser (or equivalent technique) at approximately 1 bar, as measured by a HELOS or other laser diffraction system, divided by the VMGD of the same dry powder or respirable dry particle measured by a HELOS / RODOS at 4 bar. That is, a highly dispersible dry powder or respirable dry particle will have a 1 bar / 4 bar dispersibility ratio or a 0.5 bar / 4 bar dispersibility ratio close to 1.0. Highly dispersible powders have a low tendency to agglomerate, aggregate, or conglomerate when emitted from an inhaler and inhaled by a subject, and / or, if agglomerate, aggregate, or conglomerate, are easily dispersed or deagglomerated. In another embodiment, dispersibility is assessed by measuring the particle size emitted from the inhaler as a function of flow rate. A decrease in the flow rate through the inhaler reduces the amount of energy in the airflow available to transfer to the powder to disperse it. A highly dispersible powder is one whose size distribution is characterized by a mass median aerodynamic diameter (MMAD) (aerodynamic) or VMGD (geometric) that does not increase substantially over the range of flow rates typical of human inhalation, e.g., about 15 to about 60 liters per minute (LPM), about 20 to about 60 LPM, or about 30 to about 60 LPM. A highly dispersible powder has an emitted powder mass or dose, or capsule emitted powder mass or dose, of about 80% or more even at lower inhalation flow rates. VMGD is sometimes referred to as median volume diameter (VMD), ×50, or Dv50.

[0051] The term "dry particles," as used herein, refers to respirable particles that may contain up to about 15% total water and / or other solvents. Preferably, the dry particles contain up to about 10% total water and / or other solvents by weight of the dry particles, up to about 5% total water and / or other solvents, up to about 1% total water and / or other solvents by weight of the dry particles, or 0.01% to 1% total water and / or other solvents by weight of the dry particles, or can be substantially free of water and / or other solvents.

[0052] The term "dry powder," as used herein, refers to a composition comprising respirable dry particles. The dry powder may contain up to about 15% total water and / or other solvents. Preferably, the dry powder contains up to about 10% total water and / or other solvents by weight of the dry powder, up to about 5% total water and / or other solvents, up to about 1% total water and / or other solvents by weight of the dry powder, or 0.01% to 1% total water and / or other solvents, or can be substantially free of water and / or other solvents. In one aspect, the dry powder is a respirable dry powder.

[0053] The term "effective amount," as used herein, refers to the amount of drug necessary to achieve a desired effect, e.g., treatment of cancer, e.g., lung cancer, e.g., non-small cell lung cancer. The actual effective amount for a particular application may vary depending on the particular dry powder or respirable dry particles, the mode of administration, the subject's age, weight, general health, and the severity of the symptom or condition being treated. The appropriate dosage of the dry powder or dry particles, as well as the administration schedule for a particular patient, can be determined by a clinician of ordinary skill based on these and other considerations.

[0054] As used herein, the term "emitted dose" or "ED" refers to a measure of the amount of formulation delivered from a suitable inhalation device after exhalation or dispersion has occurred. More specifically, for dry powders, ED is a measure of the percentage of powder drawn from a unit-dose package and expelled from the mouthpiece of the inhalation device. ED is defined as the ratio of the drug or powder delivered by the inhalation device to the nominal dose (i.e., the mass per unit dose of drug or powder placed into a suitable inhalation device before exhalation). ED is an experimentally measured parameter and is defined in accordance with USP Section 601 Aerosols, Metered-Dose Inhalers and Dry Powder Inhalers, Delivered-Dose Uniformity, Sampling the Delivered Dose from Dry Powder Inhalers, United States Pharmacopeia convention, Rockville, MD, 13 th Revision, 222-225, 2007, which uses an in vitro device configured to mimic patient administration. ED can also be calculated from results generated by experiments with a Next Generation Impactor (NGI) by combining all drug or powder assayed from the mouthpiece adapter, the NGI induction port, and all stages within the NGI. Results generated through ED testing according to USP 601 and via the NGI typically show good agreement.

[0055] The term "lung to plasma ratio" or "lung:plasma ratio" refers to the ratio of the concentration of an angiogenesis inhibitor (e.g., itraconazole) in the lung to the concentration of the angiogenesis inhibitor in the plasma either at a given time point or over a given range of time points. For example, the lung:plasma ratio may be calculated based on the maximum concentration of the angiogenesis inhibitor in the lung or serum (i.e., "C max") or based on concurrent measurements at any time point. The lung:plasma ratio may be calculated for total exposure (i.e., "area under the curve" or "AUC") over a specific period, such as a 24-hour period. The lung concentration of the angiogenesis inhibitor may be assessed by measuring levels in sputum, by lung lavage, by biopsy, or by some other method. The lung:plasma ratio may be calculated based on concurrent measurements at any time point in the dosing cycle, and may be calculated based on concurrent measurements before or at steady state.

[0056] The term "nominal dose," as used herein, refers to an individual dose of an angiogenesis inhibitor (e.g., itraconazole). The nominal dose is the total dose of an angiogenesis inhibitor (e.g., itraconazole) in one container, such as a capsule, blister, or ampoule.

[0057] The terms "FPF(<X)", "FPF(<X micrometers)", and "percentage of particles less than X micrometers" when used herein, where X is, for example, 3.4 micrometers, 4.4 micrometers, 5.0 micrometers, or 5.6 micrometers, refer to the percentage of dry particle samples with an aerodynamic diameter less than X micrometers. For example, FPF(<X) can be determined by dividing the mass of respirable dry particles deposited on stage 2 of a 2-stage foldable Andersen Cascade Impactor (ACI) and the final collection filter by the mass of respirable dry particles weighed into a capsule for delivery to the instrument. This parameter may be defined as "FPF_TD(<X)", where TD means total dose. Similar measurements can be made using an 8-stage ACI. The cut-off values of the 8-stage ACI are different for each at a standard flow rate of 60 L / min, but FPF_TD(<X) can be estimated from a complete dataset of the 8-stage ACI. The results of the 8-stage ACI can also be calculated by the USP method of defining FPF using the dose collected by the ACI instead of the amount placed in the capsule. Similarly, a 7-stage Next Generation Impactor (NGI) can be used.

[0058] The terms "FPD(<X)", "FPD <X micrometers", "FPD(<X micrometers)", and "particle dose less than X micrometers" when used herein, where X is, for example, 3.4 micrometers, 4.4 micrometers, 5.0 micrometers, or 5.6 micrometers, refer to the mass of therapeutic agent delivered by respirable dry particles with an aerodynamic diameter less than X micrometers. FPD <X micrometers can be determined by either directly calculating or estimating the FPD value by using an 8-stage Andersen Cascade Impactor (ACI) or Next Generation Impactor (NGI) at a standard flow rate of 60 L / min and summing the mass deposited on the final collection filter. Similarly, a 7-stage Next Generation Impactor (NGI) can be used.

[0059] The term "respirable," as used herein, refers to dry particles or dry powders suitable for delivery by inhalation to the respiratory tract (e.g., pulmonary delivery) in a subject. Respirable dry powders or respirable dry particles have a mass median aerodynamic diameter (MMAD) of less than about 10 micrometers, preferably about 5 micrometers or less.

[0060] As used herein, the term "respiratory tract" includes the upper respiratory tract (e.g., nasal passages, nasal cavities, throat, pharynx, and larynx), the respiratory airways (e.g., trachea, bronchi, and bronchioles), and the lungs (e.g., respiratory bronchioles, alveolar ducts, alveolar sacs, and alveoli).

[0061] As used herein, the term "lower respiratory tract" includes the respiratory tract and lungs.

[0062] When the term "small" is used herein to describe respirable dry particles, it refers to particles having a volume median geometric diameter (VMGD) of about 10 micrometers or less, preferably about 5 micrometers or less or less than 5 micrometers.

[0063] The term "stabilizer," as used herein, refers to a compound that improves the physical stability of an angiogenesis inhibitor in crystalline particulate form (e.g., reduces strong aggregation, weak aggregation, Ostwald ripening, and / or flocculation of the particulates) when the angiogenesis inhibitor is suspended in a liquid in which the angiogenesis inhibitor is poorly soluble. Suitable stabilizers are surfactants and amphiphiles, such as polysorbates (PS, polyoxyethylated sorbitan fatty acid esters), for example, polysorbate 20 (PS20), polysorbate 40 (PS40), polysorbate 60 (PS60) and polysorbate 80 (PS80), fatty acids, for example, lauric acid, palmitic acid, myristic acid, oleic acid and stearic acid, sorbitan fatty acid esters, for example, Span 20, Span 40, Span 60, Span 80 and Span 85, phospholipids, for example, dipalmitoylphosphatidylcholine (DPPC), 1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DSPC), 1-palmitoylphosphatidyl ... Examples of suitable stabilizers include 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), phosphatidylglycerols (PGs) such as diphosphatidylglycerol (DPPG), DSPG, DPPG, and POPG, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), aliphatic alcohols, benzyl alcohol, polyoxyethylene-9-lauryl ether, glycocholate, surfactin, poloxomers, polyvinylpyrrolidone (PVP), PEG / PPG block copolymers (Pluronics / Poloxamers), polyoxyethylene cholesteryl ether, POE alkyl ether, tyloxapol, and lecithin. Preferred stabilizers include polysorbates and fatty acids. A particularly preferred stabilizer is polysorbate 80 (PS80). Another preferred stabilizer is oleic acid or a salt thereof.

[0064] The term "homogeneous dry particles," as used herein, refers to particles that are compositionally homogeneous. Homogeneous dry particles disclosed herein have substantially the same composition of angiogenesis inhibitor (e.g., itraconazole, e.g., crystalline itraconazole), stabilizer, and optionally one or more excipients, excluding blends of two or more particles.

[0065] Therapeutic Uses and Methods The present disclosure relates to a method of treating cancer (e.g., lung cancer, e.g., NSCLC), comprising administering to the respiratory tract of a subject in need thereof a dry powder comprising homogenous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) and, optionally, a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., leucine and sodium sulfate).

[0066] The methods disclosed herein are particularly useful for treating subjects with lung cancer, e.g., non-small cell lung cancer (NSCLC). The NSCLC can include adenocarcinoma, squamous cell carcinoma, or large cell carcinoma. The subject can be chemotherapy-naive (e.g., a subject who has not previously received any chemotherapy). Alternatively, the subject can have previously received one or more cancer therapies (e.g., chemotherapy) and be resistant or refractory to the therapies. In other words, the methods disclosed herein can be used to treat resistant or refractory lung cancer. For example, the methods disclosed herein can include administering a dry powder to the airway of a subject whose cancer has not responded to one or more previous therapies or has become resistant to one or more previous therapies. The cancer treated using the methods disclosed herein can be a recurrent or recurrent cancer, e.g., a cancer that has returned after a period of remission from a previous cancer treatment. In a preferred embodiment, the subject is a human.

[0067] The methods disclosed herein can be used to treat primary or metastatic cancer. The cancer (e.g., lung cancer) can be early stage, locally advanced, or advanced. The cancer can be at any stage, including stage 0, stage I, stage II, stage III (including stages IIIA and IIIB), or stage IV (including stages IVA and IVB). For example, the methods disclosed herein can be used to treat a subject with stage 0 NSCLC, stage I NSCLC, stage II NSCLC, stage IIIA NSCLC, stage IIIB NSCLC, stage IVA NSCLC, or stage IVB NSCLC. In some embodiments, the cancer is advanced, metastatic, and / or refractory cancer (e.g., advanced NSCLC, metastatic NSCLC, and / or refractory NSCLC). In some embodiments, the methods of the present invention treat only primary cancer (e.g., when the method is not intended to treat metastatic cancer).

[0068] The cancers treated using the methods disclosed herein may be associated with one or more of an EGFR mutation (e.g., a sensitizing EGFR mutation), a KRAS mutation, an ALK rearrangement, a ROS1 rearrangement, a BRAF V600E mutation, an NTRK gene fusion, MET exon 14 skipping, a RET rearrangement, or PD-L1.

[0069] The therapeutic methods disclosed herein can inhibit, for example, cancer cell growth or tumor growth of lung cancer, e.g., NSCLC. For example, administering a therapeutically effective amount of the dry powder disclosed herein to the respiratory tract of a subject (optionally in combination with an additional therapeutic agent and / or radiation) can inhibit cell growth or tumor growth by at least about 20%, e.g., about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% or more relative to an untreated subject or a subject not treated with the dry powder disclosed herein. The therapeutic methods disclosed herein can result in tumor regression that can be observed over a period of at least about 10 days, at least about 20 days, at least about 30 days, at least about 2 months, at least about 4 months, at least about 6 months, or at least about 1 year or more.

[0070] The methods disclosed herein may extend the survival of a treated subject. For example, administering a therapeutically effective amount of the dry powder disclosed herein to the airways of a subject (optionally in combination with an additional therapeutic agent and / or radiation) can extend the survival of a subject of the invention by at least one month compared to another subject who did not receive the treatment or compared to a subject who received a treatment that did not include the dry powder disclosed herein. In some embodiments, the survival is extended by at least two months, at least three months, at least four months, at least five months, or at least six months or more.

[0071] The methods disclosed herein may extend the progression-free survival of a treated subject. For example, administering a therapeutically effective amount of the dry powder disclosed herein to the airways of a subject (optionally in combination with an additional therapeutic agent and / or radiation) may extend the progression-free survival of a subject of the invention by at least one month compared to another subject who did not receive the treatment or compared to a subject who received a treatment that did not include the dry powder disclosed herein. In some embodiments, the progression-free survival is extended by at least two months, at least three months, at least four months, at least five months, or at least six months or more.

[0072] The methods disclosed herein increase the response rate in a group of subjects compared to another group that did not receive the treatment or a group that received a treatment that did not include the dry powder disclosed herein, hi some embodiments, the response rate is increased by at least about 1%, at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, or at least about 50% or more.

[0073] The methods disclosed herein may improve symptoms of cancer (e.g., lung cancer) in a subject with cancer, e.g., lung cancer, e.g., NSCLC. For example, the methods disclosed herein may improve one or more of the following symptoms in a subject: shortness of breath or dyspnea, cough (e.g., persistent or chronic cough), blood in sputum, pain (e.g., chest pain or back pain), lung infection, jaundice, bloating or abdominal distension, headache, dizziness, seizures, lethargy or numbness (e.g., lethargy or numbness in the hands and feet), fatigue, and unexplained weight loss.

[0074] The methods disclosed herein may include administering the dry powder to the subject's respiratory tract before or after surgery to treat the cancer, such as induction therapy, neoadjuvant therapy, or adjuvant therapy. For example, the methods disclosed herein may include administering the dry powder to the subject's respiratory tract before or after surgery to remove cancerous tissue from the subject. The surgery may be any surgery typically performed to treat cancer, particularly lung cancer, such as a segmentectomy, sleeve resection, wedge resection, lobectomy, pneumonectomy, lymph node resection, or a combination thereof. For example, prior to surgery to remove cancerous tissue from the lung, the subject may be administered one or more doses of the dry powder disclosed herein (either alone or as part of a combination therapy disclosed herein) at least one day before the surgery, such as at least three days, at least one week, at least two weeks, at least three weeks, at least four weeks, at least one month, at least two months, at least three months, or at least four months or more before the surgery. The subject may be administered the dry powder in multiple doses over a period of time, for example, ending just before surgery. Alternatively or additionally, after surgery to remove cancerous tissue from the lung, the subject may be administered one or more doses of the dry powder disclosed herein (either alone or as part of a combination therapy disclosed herein) at least one day after the surgery, for example, at least three days, at least one week, at least two weeks, at least three weeks, at least four weeks, at least one month, at least two months, at least three months, or at least four months or more after the surgery. In particular, the methods disclosed herein may be used as maintenance therapy and / or to prevent disease recurrence.

[0075] Combination therapy The methods disclosed herein can include combination therapy. In particular, if desired or indicated, the dry powders described herein can be administered with one or more other therapeutic agents, such as the additional therapeutic agents disclosed herein. The other therapeutic agents can be administered by any suitable route, which routes are described in more detail below. The dry powders of the present invention can be administered before, substantially simultaneously with, or after the administration of the other therapeutic agent. Preferably, the dry powder and the additional therapeutic agent are administered so as to substantially overlap their pharmacological activities.

[0076] For example, the method may include administering the dry powder disclosed herein to a subject in addition to one or more therapeutic agents and / or radiation. The additional therapeutic agent may be any suitable agent used in the treatment of cancer, such as any chemotherapeutic agent known in the art for treating lung cancer. For example, the dry powder disclosed herein may be administered to a subject being treated with standard care treatments outlined in the National Comprehensive Cancer Network (NCCN) Clinical Practice Guidelines in Oncology. The combination therapy may include administering the dry powder of the present invention in addition to chemoradiation. The combination therapy may include administering the dry powder in addition to platinum-doublet chemotherapy. The additional therapeutic agent may be a chemotherapeutic agent, a targeted cancer therapy, an immunotherapy, or a combination thereof.

[0077] The combination therapy can include administering a dry powder disclosed herein in addition to oral itraconazole and, optionally, one or more additional therapeutic agents. For example, the methods disclosed herein can include administering a dry powder comprising itraconazole to a subject in addition to oral itraconazole and, optionally, one or more additional therapeutic agents (e.g., chemotherapeutic agents), such as standard of care treatment for lung cancer.

[0078] The additional therapeutic agent and / or oral itraconazole may be administered to the subject at, around, or before the time the dry powder is administered to the subject. For example, the additional therapeutic agent and / or oral itraconazole may be administered at a dose no more than 24 hours before or after the administration of a dose of the dry powder, for example, within about 12 hours, about 6 hours, about 4 hours, about 3 hours, about 2 hours, about 1 hour, or about 30 minutes, or less, of administration of the dry powder. However, the additional therapeutic agent and / or oral itraconazole need not be administered at, around, or before the time the dry powder is administered. For example, the dry powder may be administered to the subject more than one day before or after, e.g., two, three, four, five, six, one, two, two, three, four or more weeks before or after, the administration of the additional chemotherapeutic agent and / or oral itraconazole, depending, e.g., on the desired therapeutic outcome or the needs of the subject. In some embodiments, the dry powder is administered to the subject about 1 to about 28 days before the administration of the additional chemotherapeutic agent and / or oral itraconazole, e.g., 1 to about 21 days, about 1 to about 18 days, about 1 to about 16 days, about 1 to about 14 days, about 1 to about 12 days, or about 1 to about 10 days before the administration of the additional chemotherapeutic agent and / or oral itraconazole.

[0079] The additional therapeutic agent used in the combination therapies disclosed herein can be any suitable agent known in the art to treat cancer, e.g., lung cancer, For example, the therapeutic agent can be selected from the group consisting of platinum-based drugs, antimetabolites, anti-microtubule agents, topoisomerase inhibitors, anthracyclines, KRAS inhibitors, ALK inhibitors, EGFR inhibitors, VEGF inhibitors, BRAF inhibitors, MEK inhibitors, RET inhibitors, MET inhibitors, and immunotherapeutic agents (e.g., immune checkpoint inhibitors, e.g., PD-1 / PD-L1 inhibitors or CTLA-4 inhibitors), etc.

[0080] For example, the additional therapeutic agent can be pemetrexed, cisplatin, carboplatin, oxaliplatin, satraplatin, picoplatin, nedaplatin, triplatin, lipoplatin, paclitaxel (including albumin-bound paclitaxel), docetaxel, doxorubicin, gemcitabine, vinblastine, vinorelbine, etoposide, SN-38, camptothecin, topotecan, exatecan, irinotecan, belotecan, methotrexate, bevacizumab (AVASTIN®), ranibizumab, ramucirumab (CYRAMZA®), or ribozyme inhibitor (RIRI). )), nintedanib, aflibercept, erlotinib (TARCEVA®), afatinib (GILOTRIF®), axitinib, gefitinib (IRESSA®), cabozantinib (COMETRIQ®, CABOMETYX®), osimertinib (TAGRISSO®), dacomitinib (VIZIMPRO®), sotorasib, crizotinib (XALKORI®), entrectinib (ROZLYTREK®), lenvatinib, pazopanib, ceritinib (ZYKADIA®), alectinib (ALECENSA®), brigatinib (ALUNBRIG®), lorlatinib (LORBRENA®), dabrafenib (TAFINLAR®), regorafenib, vemurafenib (ZELBORAF®), sorafenib, sunitinib, everolimus, thalidomide, lenalidomide, trametinib (MEKINIST®), vandetanib (CAPRELSA®), selpercatinib (RETEVMO™), pralzata Mab (GAVRETO™), capmatinib (TABRECTA™), tepotinib (TEPMETKO®), larotrectinib (VITRAKVI®), amivantamab, mobocertinib, nivolumab (OPDIVO®), ipilimumab (YERVOY®), atezolizumab (TECENTRIQ®), cetuximab (ERBITUX®), pembrolizumab (KEYTRUDA®), tremelimumab, cemiplimab (LIBTAYO®), pidilizumab,It may be selected from the group consisting of durvalumab (IMFINZI®), necitumumab, oral or intravenous itraconazole, and combinations thereof.

[0081] The additional therapeutic agent may be any of the compounds described in U.S. Patent Nos. 5,997,318, 6,051,227, 6,682,736, 6,808,710, 6,984,720, 7,034,121, 7,169,901, 7,297,334, 7,423,125, 7,488,802, 7,605,238, 7,943,743, 8,008,449, 8,034,905, 8,168,757, 8,354,509, 8,609,089, 8,686,119, Nos. 8,779,105, 8,779,108, 8,900,587, U.S. Patent Application Publication Nos. US2012 / 263677, US2014 / 0356353 or US2021 / 0324106, or any one of International Publication Nos. WO2007 / 113648, WO2012 / 145493, WO2012 / 122444, WO2013 / 014668, WO2013 / 173223, each of which is incorporated herein by reference in its entirety.

[0082] Non-limiting examples of standard of care treatment regimens that may be used in the combination therapies disclosed herein (e.g., in combination with dry powders) include carboplatin or cisplatin, pemetrexed and pembrolizumab, carboplatin, paclitaxel and atezolizumab (optionally in combination with bevacizumab), carboplatin, albumin-bound paclitaxel and atezolizumab, nivolumab and ipilimumab, carboplatin or cisplatin, pemetrexed, nivolumab and ipilimumab, These include pemetrexed, cisplatin, paclitaxel, and optionally bevacizumab, carboplatin or cisplatin, pemetrexed, and optionally bevacizumab, cisplatin, and another chemotherapy, carboplatin, and another chemotherapy, gemcitabine, and docetaxel or vinorelbine, single-agent chemotherapy, carboplatin, paclitaxel, and pembrolizumab, carboplatin, albumin-bound paclitaxel, and pembrolizumab, or carboplatin, paclitaxel, nivolumab, and ipilimumab. Those skilled in the art will understand that one or more of the above therapeutic agents may be replaced or omitted by another therapeutic agent, for example, depending on the subject's condition, disease state, or response to the therapy.

[0083] The additional therapeutic agent can be administered to a subject by any suitable route of administration, including, but not limited to, oral, intravenous, intramuscular, inhalation (e.g., intrabronchial, intranasal, oral inhalation, intranasal nasal drop), intraarterial, intrathecal, intralymphatic, intralesional, intracapsular, intraorbital, intracardiac, intradermal, transtracheal, subcuticular, intraarticular, subcapsular, intrathecal, subcutaneous, intraperitoneal, intrathecal, epidural, intrasternal, intratumoral, topical, epidermal, mucosal, intranasal, vaginal, rectal, or sublingual.

[0084] The additional therapeutic agent and / or oral itraconazole can be administered to the subject at any appropriate dose, which may be a dose approved for the therapeutic agent and / or for a particular type or stage of cancer. However, it will be understood that the dosage of the compounds or compositions of the present disclosure will be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject or organism will depend on a variety of factors, including, but not limited to, the disease or disorder being treated, the severity of the disease or disorder being treated, the activity of the specific compound or composition being administered, the specific compound or composition being administered, the age, weight, general health, sex, and / or diet of the subject, the time of administration, route of administration, and excretion or metabolic rate of the specific compound or composition being administered, the duration of treatment, drugs used in combination with or concurrently with the specific compound or composition being administered, and similar factors known in the art. It will also be understood that the attending physician may determine or adjust the dosage, e.g., to reduce the dosage of the additional therapeutic agent and / or oral itraconazole when combined with the dry powder disclosed herein. Similarly, the total amount of dry powder administered, or the total drug load in the administered dry powder, may be adjusted (eg, reduced) when combined with one or more additional therapeutic agents and / or oral itraconazole.

[0085] The method for treating cancer disclosed herein may also include radiation therapy. For example, the method may include administering the dry powder disclosed herein to the airway of a subject with cancer (optionally in combination with one or more additional therapeutic agents disclosed herein) before or after administering radiation therapy to treat the cancer. The radiation therapy may be any suitable radiation therapy for treating cancer, particularly lung cancer, such as brachytherapy, external beam radiation therapy (EBRT), stereotactic body radiation therapy (SBRT), stereotactic ablative body radiation therapy (SABR), three-dimensional conformal radiation therapy (3D-CRT), intensity-modulated radiation therapy (IMRT), proton therapy, intensity-modulated arc therapy (VMAT), stereotactic radiotherapy (SRS), radiofrequency ablation, or a combination thereof.

[0086] Pharmacokinetics, Dosage, and Other Considerations The amount of dry powder administered to a subject may be sufficient to maintain a steady-state concentration. As used herein, steady-state concentration (Css) refers to the drug concentration in, for example, the lung or plasma at the point where a "steady state" is achieved and the drug administration rate and drug elimination rate are equal. The steady-state concentration is a limiting value that is theoretically reached after the end of an infinite number of equally spaced doses. The maximum value (Css,max) under such conditions is given by Css,max = C0 / (1-f) for a drug eliminated from a one-compartment system by first-order kinetics. The ratio Css,max / C0 indicates the extent to which a drug accumulates under a specific dose regimen of theoretically infinite duration, and the corresponding ratio 1 / (1-f) is sometimes referred to as the accumulation rate, and R.Css is also the limiting value theoretically reached at the "end" of a constant-rate infusion for an infinite period.

[0087] In some embodiments, an angiogenesis inhibitor, e.g., itraconazole, may be administered at a nominal dose of about 2 mg, about 3 mg, about 4 mg, 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 50 mg, about 2 mg to about 35 mg, about 5 mg to about 50 mg, about 10 mg to about 50 mg, or about 15 mg to about 50 mg. The dose and administration regimen may be selected to achieve a particular lung:plasma ratio of the angiogenesis inhibitor or to achieve a particular steady-state concentration of the angiogenesis inhibitor in the lung and / or plasma.

[0088] The lung:plasma ratio can be at least about 100:1, at least about 200:1, at least about 300:1, at least about 400:1, at least about 500:1, at least about 600:1, at least about 700:1, at least about 800:1, at least about 1000:1, at least about 1300:1, at least about 1600:1, at least about 1900:1, at least about 2200:1, at least about 2500:1, at least about 2800:1, at least about 3000:1, at least about 3200:1, at least about 3400:1, at least about 3600:1, 3000:1 to 4000:1, 3500:1 to 4000:1, or 3600:1 to 3700:1. Additionally, the lung:plasma ratio may be at least about 2:1, at least 3:1, at least 4:1, at least 5:1, at least 6:1, at least 7:1, at least 8:1, at least 9:1, at least 10:1, at least 15:1, at least 20:1, at least 25:1, at least 50:1, or at least 75:1. The lung:plasma ratio is determined based on the maximum concentration of the angiogenesis inhibitor in the lung or serum (i.e., "C max "), or at any time point. The lung:plasma ratio may be calculated for total exposure (i.e., "area under the curve" or "AUC") over a specific period of time, such as a 24 hour period. The lung:plasma ratio may be calculated based on concurrent measurements at any time point during the dosing cycle, or may be calculated before or at steady state.

[0089] At steady state, the lung:plasma ratio is at least about 20:1, at least about 25:1, at least 50:1, at least 75:1, at least about 100:1, at least about 200:1, at least about 300:1, at least about 400:1, at least about 500:1, at least about 600:1, at least about 700:1, at least about 800:1, at least about 1000:1, at least about It may be about 1300:1, at least about 1600:1, at least about 1900:1, at least about 2200:1, at least about 2500:1, at least about 2800:1, at least about 3000:1, at least about 3200:1, at least about 3400:1, at least about 3600:1, 3000:1 to 4000:1, 3500:1 to 4000:1, or 3600:1 to 3700:1.

[0090] The dry powders disclosed herein may be administered to achieve a particular plasma concentration of an angiogenesis inhibitor (e.g., itraconazole), which may be less than 40 ng / mL, less than 35 ng / mL, less than 30 ng / mL, less than 25 ng / mL, less than 20 ng / mL, less than 15 ng / mL, less than 12 ng / mL, less than 10 ng / mL, less than 8 ng / mL, less than 6 ng / mL, less than 4 ng / mL, less than 2 ng / mL, less than 1.5 ng / mL, less than 1.0 ng / mL, less than 0.5 ng / mL, less than 0.3 ng / mL, or less than 0.2 ng / mL.

[0091] The dry powder disclosed herein may be administered to achieve a specific steady-state plasma concentration of an angiogenesis inhibitor (e.g., itraconazole). At steady state, the plasma concentration may be less than 25 ng / mL, less than 20 ng / mL, less than 15 ng / mL, less than 12 ng / mL, less than 10 ng / mL, less than 8 ng / mL, less than 6 ng / mL, less than 4 ng / mL, less than 2 ng / mL, less than 1.5 ng / mL, less than 1.0 ng / mL, less than 0.5 ng / mL, less than 0.3 ng / mL, or less than 0.2 ng / mL. In addition, the steady-state plasma concentration may be less than 40 ng / mL, less than 35 ng / mL, or less than 30 ng / mL.

[0092] The lung concentration of an angiogenesis inhibitor may be determined, for example, based on the concentration of the angiogenesis inhibitor in sputum or lung tissue (e.g., lung tumor tissue). The lung concentration is determined based on the highest concentration (i.e., "C") of the angiogenesis inhibitor (e.g., itraconazole) in lung tissue or sputum. max The lung concentration may be measured at any time during the dosing cycle or calculated before or at steady state.

[0093] For example, the dry powders disclosed herein may be administered in one or more doses to increase lung concentrations (sputum concentrations) of angiogenesis inhibitors (e.g., itraconazole) to at least 100 ng / mL, e.g., about 500 ng / mL, about 800 ng / mL, about 1200 ng / mL, about 1600 ng / mL, about 2000 ng / mL, about 3000 ng / mL, about 4000 ng / mL, about 5000 ng / mL, about 6000 ng / mL, about 7000 ng / mL, about 8000 ng / mL, about 10,000 ng / mL, about 15,000 ng / mL, about 20,000 ng / mL, about 25,000 ng / mL, about 50,000 ng / mL, about 75,000 ng / mL, about 1 The concentration may be about 00,000ng / mL, about 125,000ng / mL, about 150,000ng / mL, about 175,000ng / mL or about 200,000ng / mL or more, or about 500ng / mL to 400,000ng / mL, about 500ng / mL to 300,000ng / mL, about 500ng / mL to 200,000ng / mL, about 500ng / mL to 100,000ng / mL, about 500ng / mL to 50,000ng / mL, about 500ng / mL to 25,000ng / mL, about 500ng / mL to 10,000ng / mL, about 2000ng / mL to 8000ng / mL or about 2000ng / mL to 8100ng / mL.

[0094] The dry powders disclosed herein may be administered in one or more doses to achieve a lung concentration (tissue concentration, e.g., tumor tissue concentration) of an angiogenesis inhibitor (e.g., itraconazole) of at least about 100 ng / g, e.g., about 500 ng / g, about 800 ng / g, about 1200 ng / g, about 1600 ng / g, about 2000 ng / g, about 3000 ng / g, about 4000 ng / g, about 5000 ng / g, about 6000 ng / g, about 7000 ng / g, about 8000 ng / g, about 9000 ng / g, or about 10,000 ng / g or more. In some embodiments, the dry powders disclosed herein may be administered in one or more doses to achieve lung concentrations (tissue concentrations, e.g., tumor tissue concentrations) of angiogenesis inhibitors (e.g., itraconazole) of up to about 1.6 mg / g, up to about 1.4 mg / g, up to about 1.2 mg / g, or up to about 1.0 mg / g. In some embodiments, one or more doses of the dry powder disclosed herein may be administered to achieve a lung concentration (tissue concentration, e.g., tumor tissue concentration) of an angiogenesis inhibitor (e.g., itraconazole) of about 100 ng / g to about 900,000 ng / g, e.g., about 100 ng / g to about 800,000 ng / g, about 100 ng / g to about 700,000 ng / g, about 100 ng / g to about 600,000 ng / g, about 100 ng / g to about 500,000 ng / g, about 100 ng / g to about 400,000 ng / g, about 100 ng / g to about 300,000 ng / g, about 100 ng / g to about 200,000 ng / g, or about 100 ng / g to about 100,000 ng / g.

[0095] In some embodiments, the dry powders disclosed herein are administered in one or more doses to increase lung concentrations (tissue concentrations, e.g., tumor tissue concentrations) of angiogenesis inhibitors (e.g., itraconazole) to about 100 ng / g to about 1.6 mg / g, e.g., about 500 ng / g, about 1000 ng / g, about 1500 ng / g, about 2000 ng / g, about 2500 ng / g, about 5000 ng / g, about 10,000 ng / g, about 15,000 ng / g, about 20,000 ng / g, about 25,000 ng / g, about 30,000 ng / g, about 40,000 ng / g, about 50,000 ng / g, about 60,000 ng / g, about 70,000 ng / g, , about 80,000ng / g, about 90,000ng / g, about 100,000ng / g, about 120,000ng / g, about 140,000ng / g, about 160 ,000ng / g, approx. 180,000ng / g, approx. 200,000ng / g, approx. 250,000ng / g, approx. 300,000ng / g, approx. 350,000 ng / g, about 400,000 ng / g, about 450,000 ng / g, about 500,000 ng / g, about 600,000 ng / g, about 700,000 ng / g, about 800,000 ng / g, about 900,000 ng / g, about 1 mg / g, about 1.2 mg / g, about 1.4 mg / g or about 1.6 mg / g.

[0096] Similar methods can be used to determine the lung concentration of metabolites of the angiogenesis inhibitor. For example, one or more doses of the dry powder disclosed herein, including itraconazole, can be administered to achieve a lung concentration (sputum concentration) of hydroxyitraconazole of at least 10 ng / mL, e.g., about 15 ng / mL, about 25 ng / mL, about 50 ng / mL, about 75 ng / mL, about 100 ng / mL, about 200 ng / mL, about 300 ng / mL, about 400 ng / mL, about 500 ng / mL, about 800 ng / mL, or about 900 ng / mL. The concentration may be about 00ng / mL, about 1200ng / mL, about 1600ng / mL, about 2000ng / mL, about 3000ng / mL, about 4000ng / mL, about 5000ng / mL, about 6000ng / mL, about 7000ng / mL or about 8000ng / mL or more, or about 500ng / mL to 8000ng / mL, about 2000ng / mL to 8000ng / mL or about 2000ng / mL to 8100ng / mL.

[0097] The dry powders disclosed herein, comprising itraconazole, may be administered in one or more doses to achieve a lung concentration (tissue concentration, e.g., tumor tissue concentration) of hydroxyitraconazole of at least about 10 ng / g, e.g., about 25 ng / g, about 50 ng / g, about 75 ng / g, about 100 ng / g, about 200 ng / g, about 300 ng / g, about 400 ng / g, about 500 ng / g, about 800 ng / g, about 1200 ng / g, about 1600 ng / g, about 2000 ng / g, about 3000 ng / g, about 4000 ng / g, about 5000 ng / g, about 6000 ng / g, about 7000 ng / g, about 8000 ng / g, about 9000 ng / g, or about 10,000 ng / g or more.

[0098] The dry powders disclosed herein may be administered in one or more doses to achieve a steady state lung concentration of an angiogenesis inhibitor (e.g., itraconazole) in sputum of about 500 ng / mL, about 800 ng / mL, about 1200 ng / mL, about 1600 ng / mL, about 2000 ng / mL, about 3000 ng / mL, about 4000 ng / mL, about 5000 ng / mL, about 5000 ng / mL, about 6000 ng / mL, about 7000 ng / mL, about 8000 ng / mL, about 1 The concentration may be about 0,000ng / mL, about 15,000ng / mL, about 20,000ng / mL, about 25,000ng / mL, about 50,000ng / mL, or about 75,000ng / mL or more, for example, 2000ng / mL to 100,000ng / mL, 2000ng / mL to 50,000ng / mL, 2000ng / mL to 10,000ng / mL, 2000ng / mL to 8000ng / mL, or 2000ng / mL to 8100ng / mL.

[0099] The dry powders disclosed herein can be administered in one or more doses to achieve a steady-state lung concentration of an angiogenesis inhibitor (e.g., itraconazole) in tissue (e.g., tumor tissue) of about 100 ng / g to about 1.6 mg / g, e.g., about 500 ng / g, about 1000 ng / g, about 1500 ng / g, about 2000 ng / g, about 2500 ng / g, about 5000 ng / g, about 10,000 ng / g, about 15,000 ng / g, about 20,000 ng / g, about 25,000 ng / g, about 30,000 ng / g, about 40,000 ng / g, about 50,000 ng / g, about 60,000 ng / g, about 70,000 ng / g, about 80,000 ng / g, or about 90,000 ng / g. 000ng / g, about 90,000ng / g, about 100,000ng / g, about 120,000ng / g, about 140,000ng / g, about 160,000ng / g, about 180,000ng / g, about 200,000ng / g, about 250,000ng / g, about 300,000ng / g, about 350,000ng / g, about 400,000ng / g, about 450,000ng / g, about 500,000ng / g, about 600,000ng / g, about 700,000ng / g, about 800,000ng / g, about 900,000ng / g, about 1mg / g, about 1.2mg / g, about 1.4mg / g or about 1.6mg / g.

[0100] The dry powders disclosed herein may be administered in one or more doses to achieve a steady state lung concentration of a metabolite of an angiogenesis inhibitor (e.g., hydroxyitraconazole) in sputum of about 500 ng / mL, about 800 ng / mL, about 1200 ng / mL, about 1600 ng / mL, about 2000 ng / mL, about 3000 ng / mL, about 4000 ng / mL, about 5000 ng / mL, about 5000 ng / mL, about 6000 ng / mL, about 7000 ng / mL, about 8000 ng / mL, 2000 ng / mL to 8000 ng / mL, or about 2000 ng / mL to 8100 ng / mL. The dry powders disclosed herein may be administered in one or more doses to achieve a steady state lung concentration of a metabolite of an angiogenesis inhibitor (e.g., hydroxyitraconazole) in tissue (e.g., tumor tissue) of about 10 ng / g, e.g., about 25 ng / g, about 50 ng / g, about 75 ng / g, about 100 ng / g, about 200 ng / g, about 300 ng / g, about 400 ng / g, about 500 ng / g, about 800 ng / g, about 1200 ng / g, about 1600 ng / g, about 2000 ng / g, about 3000 ng / g, about 4000 ng / g, about 5000 ng / g, about 6000 ng / g, about 7000 ng / g, about 8000 ng / g, about 9000 ng / g, or about 10,000 ng / g or more.

[0101] The dry powder disclosed herein may be administered once daily, twice daily, three times daily, once every other number of days, once every three days, or once weekly, for approximately 7 days, approximately 14 days, approximately 21 days, approximately 28 days, approximately 1 month, approximately 2 months, approximately 3 months, approximately 4 months, approximately 5 months, approximately 6 months, approximately 1 year, or continuously. In some embodiments, the dry powder is administered once daily until a steady state is achieved, and then administered less frequently for up to 6 months. In some embodiments, one or more doses required to achieve a therapeutic concentration of the angiogenesis inhibitor (e.g., itraconazole) in the lungs are administered daily until a steady state is reached, and then one or more doses are administered, for example, at a lower dose or less frequently. However, it will be understood that the total daily usage of the compounds or compositions of the present disclosure will be determined by the attending physician within the scope of sound medical judgment.

[0102] Various methods and assays for determining lung and plasma concentrations are known in the art and may be used to measure lung and plasma concentrations during and after administration of the dry powder of the present invention. For example, bioassays or HPLC may be used to measure the amount of angiogenesis inhibitor in the lung (e.g., using induced sputum, bronchial lavage, spontaneous sputum, biopsy specimens) or in tumors (e.g., using biopsy specimens) after administering the dry powder to a subject for, for example, at least 7 days, at least 14 days, at least 21 days, or at least 28 days.

[0103] The dry powder and / or respirable dry particles can be administered to the respiratory tract of a subject in need thereof using any suitable method, such as instillation techniques and / or inhalation devices, such as dry powder inhalers (DPIs) or metered dose inhalers (MDIs). A significant number of DPIs are available, for example, inhalers disclosed in U.S. Pat. Nos. 4,995,385 and 4,069,819, Spinhaler® (Fisons, Loughborough, UK), Rotahalers®, Diskhaler® and Diskus® (GlaxoSmithKline, Research Triangle Technology Park, North Carolina), FlowCaps® (Hovione, Loures, Portugal), Inhalator® (Boehringer-Ingelheim, Germany), Aerolizer® (Novartis, Switzerland), the high-tolerance, extra-high-tolerance and low-tolerance RS-01™ (a well-known type of unit-dose capsule-based dry powder inhaler manufactured by Plastiape, Italy, and described in U.S. Pat. Nos. 7,284,552 and US 2018 / 0369513), and others known to those skilled in the art.

[0104] The following scientific journal articles are incorporated by reference in their entirety for their reviews of dry powder inhaler (DPI) configurations: 1) single-dose capsule DPIs, 2) multi-dose blister DPIs, and 3) multi-dose reservoir DPIs: N. Islam, E. Gladki, "Dry powder inhalers (DPIs) - A review of device reliability and innovation," International Journal of Pharmaceuticals, 360 (2008): 1-11; H. Chystyn, "Diskus Review," International Journal of Clinical Practice, June 2007, 61, 6, 1022-1036; H. Steckel, B. Muller, "In vitro evaluation of dry powder inhalers I: drug deposition of commonly used devices," International Journal of Pharmaceuticals, 154 (1997): 19-29. Some representative capsule-based DPI units are RS-01™ (Plastiape, Italy), Turbospin® (PH&T, Italy), Brezhaler® (Novartis, Switzerland), Aerolizer (Novartis, Switzerland), Podhaler® (Novartis, Switzerland), HandiHaler® (Boehringer Ingelheim, Germany), AIR® (Civitas, Massachusetts), Dose One® (Dose One, Maine), and Eclipse® (Rhone Poulenc Rorer).Some representative unit dose DPIs are Conix® (3M, Minnesota), Cricket® (Mannkind, California), Dreamboat® (Mannkind, California), Occoris® (Team Consulting, Cambridge, UK), Solis® (Sandoz), Trivair® (Trimel Biopharma, Canada), and Twincaps® (Hovione, Loures, Portugal). Some representative blister-based DPI units are Diskus® (GlaxoSmithKline (GSK), UK), Diskhaler® (GSK), Taper Dry® (3M, Minnesota), Gemini® (GSK), Twincer® (University of Groningen, Netherlands), Aspirair® (Vectura, UK), Acu-Breathe® (Respirics, Minnesota, USA), Exubra® (Novartis, Switzerland), Gyrohaler® (Vectura, UK), Omnihaler® (Vectura, UK), Microdose® (Microdose Therapeutix, USA), Multihaler® (Cipla, India), Prohaler® (Aptar), Technohaler® (Vectura, UK) and Xcelovair® (Mylan, Pennsylvania).Some representative reservoir-based DPI units are Clickhaler® (Vectura), Next DPI® (Chiesi), Easyhaler® (Orion), Novolizer® (Meda), Pulmojet® (sanofi-aventis), Pulvinal® (Chiesi), Skyehaler® (Skyepharma), Duohaler® (Vectura), Taifun® (Akela), Flexhaler® (AstraZeneca, Sweden), Turbuhaler® (AstraZeneca, Sweden), and Twisthaler® (Merck), as well as others known to those skilled in the art.

[0105] Generally, an inhalation device (e.g., a DPI) can deliver a maximum amount of dry powder or dry particles per inhalation, which amount is related to the volume of the blister, capsule (e.g., a size 000 with a volumetric capacity of 1.37 mL, a size 00 with a volumetric capacity of 950 μL, a size 0E with a volumetric capacity of 770 μL, a size 0 with a volumetric capacity of 680 μL, a size 1 with a volumetric capacity of 480 μL, a size 2 with a volumetric capacity of 360 μL, a size 3 with a volumetric capacity of 270 μL, and a size 4 with a volumetric capacity of 200 μL), or other means for introducing the dry powder and / or respirable dry particles into the inhaler. Preferably, the blister has a volume of about 360 microliters or less, about 270 microliters or less, or more preferably, about 200 microliters or less, about 150 microliters or less, or about 100 microliters or less. Preferably, the capsule is a size 2 or 4 capsule. More preferably, the capsule is a size 3 capsule. Therefore, delivery of a desired dose or effective amount may require two or more inhalations. Preferably, each dose administered to a subject in need thereof contains an effective amount of respirable dry particles or dry powder, and each dose is administered using about four or fewer inhalations. For example, each dose of dry powder or respirable dry particles can be administered in one inhalation, or two, three, or four inhalations. The dry powder and / or respirable dry particles are preferably administered in a single breath-actuated step using a passive DPI. When using this type of device, the subject's inhalation energy disperses the respirable dry particles and draws them into the airway.

[0106] Dry powders and / or respirable dry particles suitable for use in the methods of the present invention can travel through the upper airways (i.e., oropharynx and larynx), the lower airways (which include the trachea followed by bifurcations into bronchi and bronchioles), and the terminal bronchioles (which then divide into respiratory bronchioles leading to the final respiratory zone, alveoli, or deep lung). In one embodiment of the present invention, the majority of the mass of the respirable dry particles is deposited in the deep lung. In another embodiment of the present invention, delivery is primarily to the central airways. In another embodiment, delivery is to the upper airways. In a preferred embodiment, the majority of the mass of the respirable dry particles is deposited in the conducting airways.

[0107] Because the dry powders and respirable dry particles described herein are intended to be inhaled, the present disclosure excludes the use of the dry powders to make ready-to-use dispersions, known to those skilled in the art as preparations that are completed immediately prior to use, meaning immediately prior to administration of the drug to a patient.

[0108] Dry Powders and Particles The present disclosure relates to methods of treatment comprising administering to a subject in need thereof a dry powder comprising respirable dry particles comprising an angiogenesis inhibitor, which may also include a stabilizer and / or one or more excipients.

[0109] The angiogenesis inhibitor may be a compound containing one or more 1,2,4-triazole rings, such as itraconazole. In some embodiments, the angiogenesis inhibitor is a hedgehog signaling inhibitor. In some embodiments, the angiogenesis inhibitor is a CYP51 inhibitor. In some embodiments, the dry powder comprises a drug commonly used as an antifungal agent, such as itraconazole, fluconazole, fosfluconazole, voriconazole, posaconazole, albaconazole, efinaconazole, ravuconazole, fosravuconazole, isavuconazole, or a salt thereof. In a preferred embodiment, the dry powder comprises itraconazole (e.g., crystalline itraconazole).

[0110] Alternatively, the dry powder may be selected from the group consisting of bevacizumab (AVASTIN®), axitinib (INLYTA®), cabozantinib (COMETRIQ®, CABOMETYX®), everolimus (AFINITOR®), lenalidomide (REVLIMID®), lenvatinib mesylate (LENVIMA®), nintedanib (VARGATEF®, OFEV®), pazopanib (V), and rivaroxaban. The therapeutic agent may include an angiogenesis inhibitor selected from the group consisting of thalidomide (SYNOVIR, THALOMID), vandetanib (CAPRELSA), or ziv-aflibercept (ZALTRAP). Alternatively, the dry powder comprises a compound disclosed in one of U.S. Patent Nos. 8,653,083, 9,095,589, 9,346,791, 11,028,078, or International Publication Nos. WO2013 / 155218, WO2015 / 116947, or WO2020 / 072830, each of which is incorporated herein by reference in its entirety. In a preferred embodiment, the angiogenesis inhibitor is itraconazole.

[0111] The crystallinity of the angiogenesis inhibitor (e.g., itraconazole) and the size of the primary particles of the angiogenesis inhibitor can be important for effective therapy and reduced pulmonary toxicity. For example, without wishing to be bound by any particular theory, it is believed that smaller primary particles of crystalline itraconazole dissolve in airway lining fluid more quickly than larger particles of the same crystalline form of itraconazole, in part due to increased surface area. Crystalline itraconazole is also believed to dissolve in airway lining fluid more slowly than amorphous itraconazole. Thus, the dry powders described herein can be formulated with a crystalline particulate form of the angiogenesis inhibitor (e.g., itraconazole) that results in a desired degree of crystallinity and primary particle size and can be tailored to avoid unacceptable pulmonary toxicity while achieving desired pharmacokinetic properties.

[0112] The respirable dry particles may include an angiogenesis inhibitor (e.g., itraconazole) in an amount of about 1 weight percent (wt%) to about 95 wt%. Preferably, the respirable dry particles include an amount of the angiogenesis inhibitor such that a therapeutically effective dose can be administered and maintained without the need to inhale large amounts of dry powder and without the need to inhale the dry powder too frequently, e.g., more than three times per day. For example, the respirable dry particles preferably contain about 1% by weight (wt%) to 95 wt%, about 10 wt% to 75 wt%, about 15 wt% to 75 wt%, about 25 wt% to 75 wt%, about 30 wt% to 70 wt%, about 40 wt% to 60 wt%, about 50 wt% to 95 wt%, about 50 wt% to 90 wt%, about 50 wt% to 70 wt%, about 70 wt% to 90 wt%, about 60 wt% to 80 wt%, about 20 wt%, about 30 wt%, about 40 wt%, about 50 wt%, about 60 wt%, or about 70 wt% of angiogenesis inhibitor (e.g., itraconazole). The respirable dry particles may comprise about 75% by weight (wt%), about 80% by weight, about 85% by weight, about 90% by weight, or about 95% by weight of the angiogenesis inhibitor (e.g., itraconazole). In certain embodiments, the range of the angiogenesis inhibitor in the respirable dry particles is about 40% by weight (wt%) to about 90% by weight, about 55% to about 85% by weight, about 55% to about 75% by weight, or about 65% to about 85% by weight. For example, the respirable dry particles may comprise about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, or about 60% by weight of an angiogenesis inhibitor (e.g., itraconazole). The amount (by weight) of angiogenesis inhibitor present in the respirable dry particles may also be referred to as the "drug loading."

[0113] The angiogenesis inhibitor (e.g., itraconazole) can be present in the respirable dry particles in a crystalline particulate form (e.g., nanocrystals). More specifically, in the form of primary particles having a diameter of about 50 nm to about 5,000 nm (Dv50), the angiogenesis inhibitor preferably has a crystallinity of at least 50%. For example, at any desired loading (sometimes referred to as "drug loading") of the angiogenesis inhibitor, the primary particle size can be about 100 nm, about 300 nm, about 1500 nm, about 80 nm to about 300 nm, about 80 nm to about 250 nm, about 80 nm to about 200 nm, about 100 nm to about 150 nm, about 1200 nm to about 1500 nm, about 1500 nm to about 1750 nm, about 1200 nm to about 1400 nm, or about 1200 nm to about 1350 nm (Dv50). In certain embodiments, the primary particles are about 50 nm to about 2500 nm, about 80 to 1750 nm, about 50 nm to 1000 nm, about 50 nm to 800 nm, about 50 nm to 600 nm, about 50 nm to 500 nm, about 50 nm to 400 nm, about 50 nm to 300 nm, about 50 nm to 200 nm, or about 100 nm to 300 nm. Additionally, for any desired drug loading and primary particle size, the degree of crystallinity of the angiogenesis inhibitor can be at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or about 100% crystallinity. Preferably, the angiogenesis inhibitor has a crystallinity of about 100%. In some embodiments, the administered dry powder comprises homogenous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) that is at least 50% crystallinity, e.g., 55% crystallinity, 60% crystallinity, 65% crystallinity, 70% crystallinity, 75% crystallinity, 80% crystallinity, 85% crystallinity, 90% crystallinity, 95% crystallinity, 96% crystallinity, 97% crystallinity, 98% crystallinity, 99% crystallinity, or greater than 99% crystallinity.

[0114] Crystalline microparticle forms of angiogenesis inhibitors (e.g., itraconazole) can be prepared in any desired primary particle size using any suitable method, including stabilizers if desired, such as by wet milling, jet milling, or other suitable methods.

[0115] The respirable dry particles also include a stabilizer. The stabilizer helps maintain the desired size of the crystalline particulate form of the angiogenesis inhibitor (e.g., itraconazole) during wet-milling when the feedstock is spray-dried, and also helps wetting and dispersing of the crystalline particulate suspension of the angiogenesis inhibitor and maintaining the physical stability of the suspension. Preferably, only as little stabilizer as necessary to achieve the above benefits is used. The amount of stabilizer is typically a fixed ratio relative to the amount of angiogenesis inhibitor present in the dry particles, which can range from about 1:1 (angiogenesis inhibitor:stabilizer (wt:wt)) to about 50:1 (wt:wt), with a ratio of ≥ 10:1 (10:1 or greater) being preferred. For example, the ratio of angiogenesis inhibitor to stabilizer (wt:wt) in the dry particles of the invention can be ≧10:1 (10:1 or greater), about 10:1, about 20:1, about 1:1 to about 50:1, about 10:1 to about 15:1, or about 10:1 to about 20:1. In certain embodiments, the ratio is about 5:1 to about 20:1, about 7:1 to about 15:1, or about 9:1 to about 11:1. Alternatively, the ratio of angiogenesis inhibitor to stabilizer (wt:wt) in the dry particles can be 10:1 to greater than 25:1, 11:1 to 35:1, 10.5:1 to 14.5:1, 11:1 to 31:1, greater than 12:1, 11:1 to 15:1, 11.5:1 to 14:1, 13:1 to 16:1, 15:1 to 19.5:1, 19:1 to 25:1, 20.5:1 to 23:1, or 22:1 to 32:1. Alternatively, the ratio of angiogenesis inhibitor to stabilizer (wt:wt) in the dry particles can be 11.5:1 or greater, 12:1 or greater, 14:1 or greater, 15:1 or greater, 16:1 or greater, 17:1 or greater, 18:1 or greater, 19:1 or greater, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 18:1, about 19.5:1, or about 22:1.

[0116] Additionally, the amount of stabilizer present in the dry particles of the present invention can range from about 0.05% by weight (wt%) to about 45% by weight. In certain embodiments, the range is from about 1% by weight (wt%) to about 15% by weight, from about 4% by weight to about 10% by weight, or from about 5% by weight to about 8% by weight. It is generally preferred that the respirable dry particles contain less than about 10% by weight (wt%) of stabilizer, e.g., 9% by weight or less, 8% by weight or less, 7% by weight or less, 5% by weight or less, 4% by weight or less, 3% by weight or less, 2% by weight or less, or 1% by weight or less. Alternatively, the respirable dry particles contain about 5% by weight, about 6% by weight, about 7% by weight, about 7.5% by weight, about 8% by weight, or about 10% by weight of stabilizer. A particularly preferred stabilizer for use in the dry powders described herein is polysorbate 80. Another preferred stabilizer is oleic acid or a salt thereof. In contrast to conventional dry powders, where surfactants are used to prevent crystallization from occurring in the dry powder, surfactants in the present invention are added to stabilize a colloidal suspension of a crystalline compound (e.g., an angiogenesis inhibitor) in an antisolvent.

[0117] In some embodiments, the administered dry powder comprises homogenous respirable dry particles comprising an angiogenesis inhibitor (e.g., itraconazole) and a stabilizer (e.g., polysorbate 80, or oleic acid or a salt thereof), wherein the ratio of angiogenesis inhibitor to stabilizer (wt:wt) is about 1:1 to 50:1, 10:1 or more, about 10:1, about 20:1, about 5:1 to about 20:1, about 7:1 to about 15:1, or about 9:1 to about 11:1. In some embodiments, the stabilizer is present in an amount of about 0.05% to about 45% by weight, about 4% to about 10% by weight.

[0118] The respirable dry particles also include one or more excipients in any suitable and desired amount. The dry particles can include about 10% to about 99% by weight of excipients in a total content, with about 25% to about 85% by weight or about 40% to about 55% by weight being more typical. The dry particles can include about 1%, about 2%, about 4%, about 6%, about 8%, or less than about 10% by weight of excipients in a total content. In certain embodiments, the range is about 5% to about 50%, about 15% to about 50%, about 25% to about 50%, about 5% to about 40%, about 5% to about 30%, about 5% to about 20%, or about 5% to about 15%. In other embodiments, the range of excipient is from about 1% to about 9%, from about 2% to about 9%, from about 3% to about 9%, from about 4% to about 9%, from about 5% to about 9%, from about 1% to about 8%, from about 2% to about 8%, from about 3% to about 8%, from about 4% to about 8%, from about 5% to about 8%, from about 1% to about 7%, from about 2% to about 7%, from about 3% to about 7%, from about 4% to about 7%, from about 5% to about 7%, from about 1% to about 6%, from about 2% to about 6%, from about 3% to about 6%, or from about 1% to about 5%.

[0119] In some embodiments, the dry powder administered comprises one or more excipients present in an amount of about 10% to about 99% by weight, e.g., 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the one or more excipients are present in an amount of about 5% to about 50% by weight.

[0120] Many excipients are known in the art and can be included in the dry powders and dry particles described herein. Particularly preferred pharmaceutically acceptable excipients for the dry powders and dry particles described herein include monovalent and divalent metal cation salts, sugar chains, sugar alcohols, and amino acids.

[0121] Suitable monovalent metal cation salts include, for example, sodium and potassium salts. Suitable sodium salts that can be present in the respirable dry particles of the invention include, for example, sodium chloride, sodium citrate, sodium sulfate, sodium lactate, sodium acetate, sodium bicarbonate, sodium carbonate, sodium stearate, sodium ascorbate, sodium benzoate, sodium biphosphate, sodium phosphate, sodium bisulfite, sodium borate, sodium gluconate, sodium metasilicate, and the like.

[0122] Suitable potassium salts include, for example, potassium chloride, potassium bromide, potassium iodide, potassium bicarbonate, potassium nitrite, potassium persulfate, potassium sulfite, potassium bisulfite, potassium phosphate, potassium acetate, potassium citrate, potassium glutamate, dipotassium guanylate, potassium gluconate, potassium malate, potassium ascorbate, potassium sorbate, potassium succinate, potassium sodium tartrate, and any combination thereof.

[0123] Suitable divalent metal cation salts include magnesium salts and calcium salts, such as magnesium lactate, magnesium fluoride, magnesium chloride, magnesium bromide, magnesium iodide, magnesium phosphate, magnesium sulfate, magnesium sulfite, magnesium carbonate, magnesium oxide, magnesium nitrate, magnesium borate, magnesium acetate, magnesium citrate, magnesium gluconate, magnesium maleate, magnesium sulfate, magnesium malate, magnesium taurate, magnesium orotate, magnesium glycinate, magnesium naphthenate, magnesium acetylacetonate, magnesium formate, magnesium hydroxide, magnesium stearate, magnesium hexafluorosilicate, magnesium salicylate, or any combination thereof.

[0124] Suitable calcium salts include, for example, calcium chloride, calcium sulfate, calcium lactate, calcium citrate, calcium carbonate, calcium acetate, calcium phosphate, calcium alginate, calcium stearate, calcium sorbate, calcium gluconate, and the like.

[0125] A preferred sodium salt is sodium sulfate. A preferred sodium salt is sodium chloride. A preferred sodium salt is sodium citrate. A preferred magnesium salt is magnesium lactate.

[0126] Carbohydrate excipients useful in this regard include monosaccharides, polysaccharides, sugar alcohols, dextrans, dextrins, and cyclodextrins. Representative monosaccharides include dextrose (anhydrous and monohydrate, also known as glucose and glucose monohydrate), galactose, D-mannose, sorbose, and the like. Representative disaccharides include lactose, maltose, sucrose, trehalose, and the like. Representative trisaccharides include raffinose, and the like. Other carbohydrate excipients, including dextrans, maltodextrins, and cyclodextrins, such as 2-hydroxypropyl-beta-cyclodextrin, can be used as desired. Representative sugar alcohols include mannitol, sorbitol, and the like. A preferred sugar alcohol is mannitol.

[0127] Suitable amino acid excipients include any naturally occurring amino acid that forms a powder under standard pharmaceutical processing techniques, including nonpolar (hydrophobic) and polar amino acids (uncharged, positively charged, and negatively charged amino acids), which are pharmaceutical grade and generally regarded as safe (GRAS) by the U.S. Food and Drug Administration. Representative examples of nonpolar amino acids include alanine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophan, and valine. Representative examples of polar uncharged amino acids include cysteine, glycine, glutamine, serine, threonine, and tyrosine. Representative examples of polar positively charged amino acids include arginine, histidine, and lysine. Representative examples of negatively charged amino acids include aspartic acid and glutamic acid. A preferred amino acid is leucine.

[0128] In one embodiment, respirable dry particles of the invention include leucine as one of the one or more excipients in an amount of about 1% to about 9%, about 2% to about 9%, about 3% to about 9%, about 4% to about 9%, about 5% to about 9%, about 1% to about 8%, about 2% to about 8%, about 3% to about 8%, about 4% to about 8%, about 5% to about 8%, about 1% to about 7%, about 2% to about 7%, about 3% to about 7%, about 4% to about 7%, about 5% to about 7%, about 1% to about 6%, about 2% to about 6%, about 3% to about 6%, about 1% to about 5%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 9%, or about 10%. In another embodiment, the respirable dry particles include leucine as one of the one or more excipients in an amount of 10% or greater.

[0129] Without wishing to be bound by theory, it is believed that combining an angiogenesis inhibitor in a dry powder with an amino acid, such as leucine, and optionally one or more excipients, such as a monovalent metal cation (e.g., a sodium salt, e.g., sodium chloride or sodium sulfate), and optionally a stabilizer (e.g., polysorbate 80, oleic acid or its salts), can provide an optimal dissolution rate for achieving an effective therapeutic level of the angiogenesis inhibitor in the lungs without unacceptable toxicity. In addition, maintaining a relatively high drug loading rate of the angiogenesis inhibitor (e.g., 40% or 50% or more) can prevent rapid dissolution of the dry powder in the lungs. For example, the dry powder disclosed herein may dissolve more slowly in the lungs than a formulation containing a relatively low amount of itraconazole (e.g., less than 40 wt%) combined with a hydrophilic excipient, such as mannitol.

[0130] Dissolution of dry powders used in the methods disclosed herein may be measured in terms of dissolution half-life. In some embodiments, the dry powders used in the methods disclosed herein have a dissolution half-life of at least about 2 minutes, e.g., about 2 to about 60 minutes, about 2 to about 40 minutes, about 2 to about 30 minutes, about 3 to about 25 minutes, about 4 to about 20 minutes, about 4 to about 18 minutes, about 2 to about 10 minutes, about 10 to about 20 minutes, about 4 to about 5 minutes, about 5 to about 6 minutes, or about 7 to about 8 minutes, e.g., about 2 minutes, about 3 minutes, about 4 minutes, about 5 minutes, about 6 minutes, about 7 minutes, about 8 minutes, about 9 minutes, about 10 minutes, about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes, about 16 minutes, about 18 minutes, about 20 minutes, about 30 minutes, or about 40 minutes. In some embodiments, the dissolution half-life is about 4.1 minutes, about 4.2 minutes, about 4.3 minutes, or about 4.4 minutes. In some embodiments, the dissolution half-life is about 7.2 minutes, about 7.3 minutes, about 7.4 minutes, or about 7.5 minutes. In some embodiments, the dissolution half-life is about 16.6 minutes, about 16.7 minutes, about 16.8 minutes, or about 16.9 minutes. In some embodiments, the dissolution half-life is about 4.13 minutes to about 16.84 minutes.

[0131] The dry particles described herein include an angiogenesis inhibitor (e.g., itraconazole, e.g., itraconazole in crystalline microparticle form) and, optionally, a stabilizer and / or one or more excipients. In some embodiments, the dry particles include a first excipient that is a monovalent or divalent metal cation salt and a second excipient that is an amino acid, a sugar chain, or a sugar alcohol. For example, the first excipient can be a sodium salt or a magnesium salt, and the second excipient can be an amino acid (e.g., leucine). In a more specific example, the first excipient can be sodium sulfate, sodium chloride, or magnesium lactate, and the second excipient can be leucine. More specifically, the first excipient can be sodium sulfate, and the second excipient can be leucine. In another example, the first excipient can be a sodium salt or a magnesium salt, and the second excipient can be a sugar alcohol (e.g., mannitol). In a more specific example, the first excipient can be sodium sulfate, sodium chloride, or magnesium lactate, and the second excipient can be mannitol. In another example, the first excipient can be a sodium salt or a magnesium salt, and the second excipient can be a sugar chain (e.g., maltodextrin). In another example, the dry particles of the present invention comprise an angiogenesis inhibitor in crystalline microparticle form, a stabilizer, and one excipient, such as a sodium salt, a magnesium salt, or an amino acid (e.g., leucine).

[0132] In one embodiment, the dry powder comprises respirable dry particles that include 1) an angiogenesis inhibitor in crystalline particulate form, 2) a stabilizer, and 3) one or more excipients.

[0133] In one preferred embodiment, the dry powder comprises respirable dry particles comprising 1) itraconazole in crystalline microparticle form, 2) a stabilizer, and 3) one or more excipients.

[0134] In one embodiment, the dry powder comprises: (i) about 50% to about 80% of an angiogenesis inhibitor (e.g., itraconazole) in crystalline microparticle form, about 4% to about 40% of a stabilizer, and about 1% to about 9% of one or more excipients; (ii) about 45% to about 85% of an angiogenesis inhibitor (e.g., itraconazole) in crystalline microparticle form, about 3% to about 15% of a stabilizer, about 3% to about 40% of a sodium salt, and about 1% to about 9% of one or more amino acids; (iii) about 45% to about 85% of an angiogenesis inhibitor (e.g., itraconazole) in crystalline microparticle form, about 3% to about 15% of a stabilizer, about 3% to about 50% of sodium sulfate, and about 1% to about 9% of one or more amino acids; (iv) about 45% to about 85% of an angiogenesis inhibitor (e.g., itraconazole) in crystalline microparticle form, about 3% to about 15% of a stabilizer, about 3% to about 50% of a sodium salt, and about 1% to about 8% of one or more amino acids; or (v) about 45% to about 85% of an angiogenesis inhibitor (e.g., itraconazole) in crystalline microparticle form, about 3% to about 15% of a stabilizer, about 3% to about 40% of sodium sulfate, and about 1% to about 8% of leucine, all percentages being by weight and all formulations adding up to 100% on a dry basis.

[0135] In a particularly preferred embodiment, the dry powder comprises respirable dry particles comprising 1) itraconazole in crystalline microparticle form, 2) a stabilizer, and 3) one or more excipients. In this particularly preferred embodiment, the dry powder does not contain lactose. A specific formulation of this particularly preferred embodiment is set forth below. In Tables 1 and 1A below, these examples are further specified for itraconazole in crystalline microparticle form with a given itraconazole crystal size (also referred to as itraconazole primary particles).

[0136] In one embodiment, the dry powder comprises 50% itraconazole, 35% sodium sulfate, 10% leucine, and 5% polysorbate 80.

[0137] In one embodiment, the dry powder comprises 50% itraconazole, 37% sodium sulfate, 8% leucine, and 5% polysorbate 80.

[0138] In another embodiment, the dry powder comprises 60% itraconazole, 26% sodium sulfate, 8% leucine, and 6% polysorbate 80.

[0139] In another embodiment, the dry powder comprises 70% itraconazole, 15% sodium, 8% leucine, and 7% polysorbate 80.

[0140] In another embodiment, the dry powder comprises 75% itraconazole, 9.5% sodium sulfate, 8% leucine, and 7.5% polysorbate 80.

[0141] In another embodiment, the dry powder comprises 80% itraconazole, 4% sodium sulfate, 8% leucine, and 8% polysorbate 80.

[0142] In another embodiment, the dry powder comprises 80% itraconazole, 10% sodium sulfate, 2% leucine, and 8% polysorbate 80.

[0143] In another embodiment, the dry powder comprises 80% itraconazole, 11% sodium sulfate, 1% leucine, and 8% polysorbate 80.

[0144] In one embodiment, the dry powder comprises 50% itraconazole, 35% sodium chloride, 10% leucine, and 5% polysorbate 80.

[0145] In one embodiment, the dry powder comprises 50% itraconazole, 37% sodium chloride, 8% leucine, and 5% polysorbate 80.

[0146] In another embodiment, the dry powder comprises 60% itraconazole, 26% sodium chloride, 8% leucine, and 6% polysorbate 80.

[0147] In another embodiment, the dry powder comprises 70% itraconazole, 15% sodium chloride, 8% leucine, and 7% polysorbate 80.

[0148] In another embodiment, the dry powder comprises 75% itraconazole, 9.5% sodium chloride, 8% leucine, and 7.5% polysorbate 80.

[0149] In another embodiment, the dry powder comprises 80% itraconazole, 4% sodium chloride, 8% leucine, and 8% polysorbate 80.

[0150] In another embodiment, the dry powder comprises 80% itraconazole, 10% sodium chloride, 2% leucine, and 8% polysorbate 80.

[0151] In another embodiment, the dry powder comprises 80% itraconazole, 11% sodium chloride, 1% leucine, and 8% polysorbate 80.

[0152] The dry powders and / or respirable dry particles are preferably small, mass-dense, and dispersible. Laser diffraction systems, such as the Spraytec system (particle size analyzer, Malvern Instruments) and the HELOS / RODOS system (laser diffraction sensor and dry dispersion unit, Sympatec GmbH), may be used to measure the volume median geometric diameter (VMGD). Respirable dry particles of the present invention have a VMGD of about 10 micrometers or less, about 5 micrometers or less, about 4 μm or less, about 3 μm or less, about 1 μm to about 5 μm, about 1 μm to about 4 μm, about 1.5 μm to about 3.5 μm, about 2 μm to about 5 μm, about 2 μm to about 4 μm, or about 2 μm to about 3 μm, as measured by laser diffraction using a HELOS / RODOS system at a dispersion pressure setting of 1.0 bar at maximum orifice ring pressure (also referred to as regulated pressure). Preferably, the VMGD is about 5 micrometers or less or about 4 μm or less. In one embodiment, the dry powder and / or respirable dry particles have a minimum VMGD of about 0.5 micrometers or about 1.0 micrometers.

[0153] The dry powder and / or respirable dry particles preferably have a dispersibility ratio at 1 bar / 4 bar and / or a dispersibility ratio at 0.5 bar / 4 bar of less than about 2.0 (e.g., from about 0.9 to about 2), or less than about 1.7 (e.g., from about 0.9 to about 1.7), or less than about 1.5 (e.g., from about 0.9 to about 1.5), or less than about 1.4 (e.g., from about 0.9 to about 1.4), or less than about 1.3 (e.g., from about 0.9 to about 1.3), and preferably at 1 bar / 4 bar and / or 0.5 bar / 4 bar of less than about 1.5 (e.g., from about 1.0 to about 1.5) and / or less than about 1.4 (e.g., from about 1.0 to about 1.4).

[0154] The dry powder and / or respirable dry particles preferably have a tap density of at least about 0.2 g / cm 3 , at least about 0.25 g / cm 3 , a tap density of at least about 0.3 g / cm 3 , at least about 0.35 g / cm 3 , tap density of at least 0.4 g / cm 3 For example, the dry powder and / or respirable dry particles may have a tap density of 0.4 g / cm 3 Over (e.g., 0.4 g / cm 3 ~Approx. 1.2g / cm 3 and a tap density of at least about 0.45 g / cm 3 (For example, about 0.45 g / cm 3 ~Approx. 1.2g / cm 3 ), at least about 0.5 g / cm 3 (For example, about 0.5 g / cm 3 ~Approx. 1.2g / cm 3 ), at least about 0.55 g / cm 3 (For example, about 0.55 g / cm 3 ~Approx. 1.2g / cm 3 ), at least about 0.6 g / cm 3 (For example, about 0.6 g / cm 3 ~Approx. 1.2g / cm 3 ) or at least about 0.6 g / cm 3 ~Approx. 1.0g / cm 3 Alternatively, the dry powder and / or respirable dry particles preferably have a tap density of about 0.01 g / cm3 ~about 0.5g / cm 3 , about 0.05g / cm 3 ~about 0.5g / cm 3 , about 0.1g / cm 3 ~about 0.5g / cm 3 , about 0.1g / cm 3 ~ approx. 0.4g / cm 3 or approximately 0.1 g / cm 3 ~ approx. 0.4g / cm 3 Alternatively, the dry powder and / or respirable dry particles have a tap density of about 0.15 g / cm 3 ~Approx. 1.0g / cm 3 Alternatively, the dry powder and / or respirable dry particles have a tap density of about 0.2 g / cm 3 ~about 0.8g / cm 3 is.

[0155] The dry powder and / or respirable dry particles have a bulk density of at least about 0.1 g / cm 3 or at least about 0.8 g / cm 3 For example, the dry powder and / or respirable dry particles have a bulk density of about 0.1 g / cm 3 ~about 0.6g / cm 3 , about 0.2g / cm 3 ~about 0.7g / cm 3 , about 0.3g / cm 3 ~about 0.8g / cm 3 is.

[0156] The respirable dry particles, and when the dry powder is a respirable dry powder, the dry powder preferably has an MMAD of less than 10 micrometers, and preferably an MMAD of about 5 micrometers or less or about 4 micrometers or less. In one embodiment, the respirable dry powder and / or respirable dry particles preferably have a minimum MMAD of about 0.5 micrometers or about 1.0 micrometer. In one embodiment, the respirable dry powder and / or respirable dry particles preferably have a minimum MMAD of about 2.0 micrometers, about 3.0 micrometers, or about 4.0 micrometers.

[0157] The dry powder and / or respirable dry particles preferably have at least about 35%, preferably at least about 45%, at least about 60%, about 45% to about 80%, or about 60% to about 80% of the total dose having an FPF of less than about 5.6 micrometers (FPF<5.6 μm).

[0158] The dry powder and / or respirable dry particles preferably have at least about 20%, preferably at least about 25%, at least about 30%, at least about 40%, about 25% to about 60%, or about 40% to about 60% of the total dose having an FPF of less than about 3.4 micrometers (FPF<3.4 μm).

[0159] The dry powders and / or respirable dry particles preferably have a total water and / or solvent content of at most about 15% by weight, at most about 10% by weight, at most about 5% by weight, at most about 1% or from about 0.01% to about 1%, or may be substantially free of water or other solvents.

[0160] The dry powder and / or respirable dry particles are preferably administered with low inhalation energy. To correlate the dispersion of powder at different inhalation flow rates, inhalation volumes, and from inhalers with different resistances, the energy required to perform the inhalation strategy can be calculated. The inhalation energy is given by E=R 2 Q 2 V, where E is the inhaled energy in joules and R is kPa 1 / 2 where Q is the inhaler resistance in L / min, Q is the steady state flow rate in L / min, and V is the inspired air volume in L.

[0161] 0.02kPa 1 / 2 / LPM and 0.055kPa 1 / 2Based on both the FDA guidance document for dry powder inhalers and a study by Tiddens et al. (Journal of Aerosol Med, 19(4), p.456-465, 2006), which found that the average adult inhalation volume across a range of DPIs was 2.2 L, along with peak inspiratory flow rates (PIFR) measured by Clarke et al. (Journal of Aerosol Med, 6(2), p.99-110, 1993) for flow rates Q from two inhaler resistances of 1 / LPM, it is predicted that using an inhalation volume of 2 L, a healthy adult population could achieve inhalation energies ranging from 2.9 Joules at a comfortable inhalation to 22 Joules at a maximal inhalation.

[0162] It is predicted that adult patients with mild COPD will have a maximum inhalation energy of 5.1 to 21 joules, those with moderate COPD will have a maximum inhalation energy of 5.2 to 19 joules, and those with severe COPD will have a maximum inhalation energy of 2.3 to 18 joules. This is also based on using the PIFR measurement in the equation for inhalation energy in relation to flow rate Q. The PIFR achievable in each group is a function of the inhaler resistance during inhalation. Using the study by Broeders et al. (Eur Respir J, 18, p. 780-783, 2001), a resistance of 0.021 kPa was calculated for each group. 1 / 2 / LPM and 0.032kPa 1 / 2 The maximum and minimum achievable PIFR values ​​for the two dry powder inhalers were predicted.

[0163] Similarly, adult asthmatics are predicted to be able to achieve a maximum inhalation energy of 7.4 to 21 joules, based on the same assumptions as the COPD population and the PIFR data from Broeders et al.

[0164] Healthy adults and children, for example, are capable of providing sufficient inhalation energy to disperse the dry powders of the present disclosure, for example, from a suitable inhalation device (e.g., a dry powder inhaler). It is expected that most cancer patients, e.g., lung cancer, e.g., NSCLC patients, will also be capable of providing sufficient inhalation energy to disperse the dry powders of the present disclosure, for example, from a suitable inhalation device.

[0165] The dry powders and / or respirable dry particles useful in the methods disclosed herein are preferably characterized by high emissions, e.g., CEPM, of at least 75%, at least 80%, at least 85%, at least 90%, or at least 95%, from the passive dry powder inhaler for a subject applying about 5 Joules, about 3.5 Joules, about 2.4 Joules, about 2 Joules, about 1 Joule, about 0.8 Joules, about 0.5 Joules, or about 0.3 Joules of total inhalation energy to the dry powder inhaler. The dry powder and / or respirable dry particle container may contain about 5 mg, about 7.5 mg, about 10 mg, about 15 mg, about 20 mg, or about 30 mg. In one embodiment, the dry powder and / or respirable dry particles are characterized by a CEPM of 80% or greater and a VMGD of 5 micrometers or less when emitted from a passive dry powder inhaler having a resistance of about 0.036 sqrt (kPa) / liter per minute using a No. 3 capsule containing 10 mg of total mass at an airflow rate of 30 LPM for 3 seconds. In another embodiment, the dry powder and / or respirable dry particles are characterized by a CEPM of 80% or greater and a VMGD of 5 micrometers or less when emitted from a passive dry powder inhaler having a resistance of about 0.036 sqrt (kPa) / liter per minute using a No. 3 capsule containing 10 mg of total mass at an airflow rate of 20 LPM for 3 seconds. In a further aspect, the dry powder and / or respirable dry particles are characterized by a CEPM of 80% or greater and a VMGD of 5 micrometers or less when emitted from a passive dry powder inhaler having a resistance of about 0.036 sqrt (kPa) / liter per minute using a No. 3 capsule containing 10 mg total mass at an airflow rate of 15 LPM for 4 seconds.

[0166] The dry powder can be filled into a unit-dose container, or the unit-dose container can be at least 2% full, at least 5% full, at least 10% full, at least 20% full, at least 30% full, at least 40% full, at least 50% full, at least 60% full, at least 70% full, at least 80% full, or at least 90% full. The unit-dose container can be a capsule (e.g., #000 with a volume capacity of 1.37 mL, #00 with a volume capacity of 950 μL, #0E with a volume capacity of 770 μL, #0 with a volume capacity of 680 μL, #1 with a volume capacity of 480 μL, #2 with a volume capacity of 360 μL, #3 with a volume capacity of 270 μL, and #4 with a volume capacity of 200 μL). The capsule can be at least about 2% full, at least about 5% full, at least about 10% full, at least about 20% full, at least about 30% full, at least about 40% full, or at least about 50% full. The unit dose container can be a blister. The blister can be packaged as a single blister or as part of a series of blisters, e.g., 7 blisters, 14 blisters, 28 blisters, or 30 blisters. One or more of the blisters can preferably be at least 30% full, at least 50% full, or at least 70% full.

[0167] An advantage of the dry powders disclosed herein is that they disperse well over a wide range of flow rates and are relatively flow rate independent, making the dry powders and / or respirable dry particles a simple passive DPI available for a wide range of patient populations.

[0168] In certain embodiments, the dry powders and / or respirable dry particles of the invention comprise an angiogenesis inhibitor in crystalline microparticle form (also referred to as crystalline primary particles of the angiogenesis inhibitor) (e.g., having a primary particle size of about 80 nm to about 1750 nm, e.g., about 60 nm to about 175 nm, about 150 nm to about 400 nm, or about 1200 nm to about 1750 nm), a stabilizer, and optionally one or more excipients. Particular dry powders and respirable dry particles have the following formulations, shown in Table 1:

[0169] Dry powders and / or respirable dry particles useful in the methods described herein are preferably characterized by: 1) a VMGD at 1 bar of about 10 micrometers or less, preferably about 5 micrometers or less, as measured using a HELOS / RODOS system; 2) a dispersibility ratio at 1 bar / 4 bar and / or a dispersibility ratio at 0.5 bar / 4 bar of about 1.5 or less, about 1.4 or less, or about 1.3 or less; 3) a MMAD of about 10 micrometers or less, preferably about 5 micrometers or less; 4) at least about 45% or at least about 60% of the total dose having an FPF<5.6 μm; and / or 5) at least about 25% or at least about 40% of the total dose having an FPF<3.4 μm. Desirably, the dry powders and / or respirable dry particles have a tap density of about 0.2 g / cm. 3 More than about 0.3g / cm 3 More than about 0.4g / cm 3 More than 0.4g / cm 3 Super, about 0.45g / cm 3 More than or about 0.5g / cm 3 The above is a further feature.

[0170] Exemplary dry powders that can be used in the methods disclosed herein are shown in Table 1 below. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0171] In certain embodiments, Formulation XII has an FPF of 57% of the total dose less than 5 microns, resulting in a sub-5 micron fine particle dose of 2.8 mg for a 10.0 mg total dry powder capsule fill.

[0172] The dry powders and / or respirable dry particles described by any of the ranges, or the specifically disclosed formulations (characterized in the paragraphs above), may be filled into a container, such as a capsule or blister. When the container is a capsule, the capsule may be, for example, a No. 2 or No. 3 capsule, preferably a No. 3 capsule. The capsule material may be, for example, gelatin or HPMC (hydroxypropyl methylcellulose), preferably HPMC.

[0173] The dry powders and / or respirable dry particles described and characterized above may be contained in a dry powder inhaler (DPI). The DPI may be a capsule-based DPI or a blister-based DPI, preferably a capsule-based DPI. More preferably, the dry powder inhaler is selected from the RS01™ series of dry powder inhalers (Plastiape SpA, Italy). More preferably, the dry powder inhaler is selected from the RS01™ HR or RS01™ UHR2. Most preferably, the dry powder inhaler is an RS01™ HR.

[0174] The dry powders used in the methods disclosed herein may comprise homogenous respirable dry particles comprising itraconazole in crystalline particulate form, polysorbate 80, and or additional excipients (e.g., monovalent metal cation salts, e.g., sodium salts), wherein the ratio of itraconazole to polysorbate 80 (wt:wt) in the dry powder is greater than 10:1, provided that the dry powder is a mixture of 20% itraconazole, 39% sodium sulfate, 39% mannitol, and 2% polysorbate 80, 50% itraconazole, 22.5% sodium sulfate, mannitol, and 1% polysorbate 80. 22.5% ethanol and 5% polysorbate 80, 20% itraconazole, 62.4% sodium chloride, 15.6% leucine and 2% polysorbate 80, 50% itraconazole, 36% sodium sulfate, 9% leucine and 5% polysorbate 80, 20% itraconazole, 66.3% magnesium lactate, 11.7% leucine and 2% polysorbate 80, 50% itraconazole, 38.25% magnesium lactate, 6.75% leucine and 5% polysorbate 80, 5% itraconazole 0%, sodium sulfate 35%, leucine 10% and polysorbate 80 5%, itraconazole 50%, sodium sulfate 35%, leucine 10% and less than 5% polysorbate 80, itraconazole 50%, sodium sulfate 35%, leucine 13.75% and polysorbate 80 1.25%, itraconazole 50%, sodium sulfate 37%, leucine 8% and polysorbate 80 5%, itraconazole 60%, sodium sulfate 26%, leucine 8% and polysorbate 80 6%, itraconazole 70% leucine, 15% sodium, 8% leucine and 7% polysorbate 80, 75% itraconazole, 9.5% sodium sulfate, 8% leucine and 7.5% polysorbate 80, 80% itraconazole, 4% sodium sulfate, 8% leucine and 8% polysorbate 80, 80% itraconazole, 10% sodium sulfate, 2% leucine and 8% polysorbate 80, or 80% itraconazole, 11% sodium sulfate, 1% leucine and 8% polysorbate 80.

[0175] Dry powders useful in the methods disclosed herein may comprise homogenous respirable dry particles comprising itraconazole in crystalline particulate form, polysorbate 80, and one or more excipients (e.g., monovalent metal cation salts, e.g., sodium salts), wherein the ratio of itraconazole to polysorbate 80 (wt:wt) in the stock solution used to prepare the dry powder is greater than 10:1, provided that the dry powder is 20% itraconazole, 39% sodium sulfate, 39% mannitol, and 2% polysorbate 80, 50% itraconazole, 50% sodium sulfate, 50% mannitol, and 2% polysorbate 80. 22.5% thorium, 22.5% mannitol and 5% polysorbate 80, 20% itraconazole, 62.4% sodium chloride, 15.6% leucine and 2% polysorbate 80, 50% itraconazole, 36% sodium sulfate, 9% leucine and 5% polysorbate 80, 20% itraconazole, 66.3% magnesium lactate, 11.7% leucine and 2% polysorbate 80, 50% itraconazole, 38.25% magnesium lactate, 6.75% leucine and 5% polysorbate 80, itraconazole Itraconazole 50%, sodium sulfate 35%, leucine 10% and polysorbate 80 5%, itraconazole 50%, sodium sulfate 35%, leucine 10% and less than 5% polysorbate 80, itraconazole 50%, sodium sulfate 35%, leucine 13.75% and polysorbate 80 1.25%, itraconazole 50%, sodium sulfate 37%, leucine 8% and polysorbate 80 5%, itraconazole 60%, sodium sulfate 26%, leucine 8% and polysorbate 80 6%, itra Provided that it does not contain 70% conazole, 15% sodium, 8% leucine and 7% polysorbate 80, 75% itraconazole, 9.5% sodium sulfate, 8% leucine and 7.5% polysorbate 80, 80% itraconazole, 4% sodium sulfate, 8% leucine and 8% polysorbate 80, 80% itraconazole, 10% sodium sulfate, 2% leucine and 8% polysorbate 80, or 80% itraconazole, 11% sodium sulfate, 1% leucine and 8% polysorbate 80.

[0176] Exemplary formulations that may be used in the methods described herein include, but are not limited to, the following: [Table 2-1] [Table 2-2] [Table 2-3]

[0177] Methods for preparing dry powders and dry particles Respirable dry particles and dry powders for use in the methods disclosed herein can be prepared using any suitable method, provided that the dry powders are not readily dispersible. Many suitable methods for preparing dry powders and / or respirable dry particles are conventional in the art and include single emulsion solvent evaporation, double emulsion solvent evaporation, spray drying, spray freeze drying, milling (e.g., jet milling), blending, solvent extraction, solvent evaporation, phase separation, simple coacervation, complex coacervation, interfacial polymerization, suitable methods involving the use of supercritical carbon dioxide (CO), ultrasonic crystallization, nanoparticle agglomerate formation, and other suitable methods, including combinations thereof. Respirable dry particles can be prepared using methods known in the art for preparing microspheres or microcapsules. These methods can be used under conditions that result in the formation of respirable dry particles with desired aerodynamic properties (e.g., aerodynamic diameter and geometric diameter). If desired, respirable dry particles with desired properties, such as size and density, can be selected using suitable methods, such as sieving.

[0178] Suitable methods for selecting respirable dry particles with desired properties, such as size and density, include wet sieving, dry sieving, and aerodynamic classifiers (eg, cyclones).

[0179] The respirable dry particles of the present invention are preferably spray-dried. Suitable spray-drying techniques are described, for example, in K. Masters' "Spray Drying Handbook" (John Wiley & Sons, New York (1984)). Generally, during spray drying, heat from a hot gas, such as heated air or heated nitrogen, is used to evaporate solvent from droplets formed by atomizing a continuous liquid feed. When hot air is used, moisture in the air is at least partially removed before its use. When nitrogen is used, the nitrogen gas can be flowed "dry," meaning that no additional water vapor is combined with the gas. If desired, the moisture level of the nitrogen or air can be set to a constant value higher than that of the "dry" nitrogen before the start of the spray-drying operation. If desired, the spray-drying apparatus or other apparatus, such as a jet mill apparatus, used to prepare the dry particles of the present invention can include an in-line geometric particle size measuring device to determine the geometric diameter of the respirable dry particles as they are generated, and / or an in-line aerodynamic particle size measuring device to determine the aerodynamic diameter of the respirable dry particles as they are generated.

[0180] In spray drying, a solution, emulsion, or suspension containing the dry particle components to be produced in a suitable solvent (e.g., aqueous solvent, organic solvent, aqueous-organic mixture, or emulsion) is sprayed into a drying vessel by an atomizing device. For example, a nozzle or rotary atomizer may be used to spray the solution or suspension into the drying vessel. The nozzle may be a two-fluid nozzle, and the nozzle may be in an internal or external mixing configuration. Alternatively, a rotary atomizer with 4 or 24 impellers may be used. Examples of suitable spray dryers that can be equipped with rotary atomizers and / or nozzles include the Mobile Minor Spray Dryer or Model PSD-1 (both manufactured by GEA Niro, Inc., Denmark), the Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), and the ProCepT Formatrix R&D spray dryer (ProCepT nv, Zelzate, Belgium), among several other spray dryer options. Actual spray drying conditions will vary, in part, depending on the composition of the solution or suspension being spray dried and the material flow rate. Those skilled in the art will be able to determine appropriate conditions based on the composition of the solution, emulsion, or suspension to be spray dried, the desired particle characteristics, and other factors. Typically, the spray dryer inlet temperature is about 90°C to about 300°C. The spray dryer outlet temperature will vary depending on factors such as the feed temperature and the properties of the material being dried. Typically, the outlet temperature is from about 50° C. to about 150° C. If desired, the respirable dry particles produced can be fractionated by volumetric size, e.g., using sieves, separated by aerodynamic size, e.g., using cyclones, and / or further separated according to density using techniques known to those skilled in the art.

[0181] To prepare the respirable dry particles of the present invention, generally, an emulsion or suspension containing the desired components (i.e., raw materials) of the dry powder of the present invention is prepared and spray-dried under appropriate conditions. Preferably, the raw materials have a dissolved or suspended solids concentration of at least about 1 g / L, at least about 2 g / L, at least about 5 g / L, at least about 10 g / L, at least about 15 g / L, at least about 20 g / L, at least about 30 g / L, at least about 40 g / L, at least about 50 g / L, at least about 60 g / L, at least about 70 g / L, at least about 80 g / L, at least about 90 g / L, or at least about 100 g / L. The raw materials can be produced by dissolving, suspending, or emulsifying the appropriate components (e.g., salts, excipients, other active ingredients) in a suitable solvent to prepare a solution, suspension, or emulsion. The solution, emulsion, or suspension can be prepared using any suitable method, such as bulk mixing of dry and / or liquid components or static mixing of liquid components to form a combination. For example, a hydrophilic component (e.g., an aqueous solution) and a hydrophobic component (e.g., an organic solution) can be combined using a static mixer to form a combination. The combination can then be atomized to produce droplets, which can be dried to form respirable dry particles. Preferably, the atomization step occurs immediately after the components are combined in the static mixer. Alternatively, the atomization step occurs on a bulk-mixed solution.

[0182] The raw material can be prepared using any solvent that the microparticle form of angiogenesis inhibitor has low solubility, such as organic solvent, aqueous solvent or their mixture.Suitable organic solvents that can be used include, but are not limited to, alcohols, such as ethanol, methanol, propanol, isopropanol, butanol, etc.Other organic solvents include, but are not limited to, tetrahydrofuran (THF), perfluorocarbons, dichloromethane, chloroform, ether, ethyl acetate, methyl tert-butyl ether, etc.Cosolvents that can be used include, but are not limited to, aqueous solvents and organic solvents (such as, but not limited to, the organic solvents mentioned above).Aqueous solvents include water and buffer solutions.Preferred solvent is water.

[0183] Various methods (e.g., static mixing, bulk mixing) can be used to mix the solute and solvent to prepare the feedstock, and are known in the art. Other suitable methods for mixing may be used if desired. For example, additional ingredients can be included in the feedstock to cause or enhance the mixing. For example, carbon dioxide can act to promote physical mixing of the solute and solvent by creating effervescence.

[0184] The raw material or raw material components can have any desired pH, viscosity, or other properties. If desired, a pH buffer can be added to the solvent or cosolvent, or to the resulting mixture. Generally, the pH of the mixture ranges from about 3 to about 8.

[0185] After the dry powder and / or respirable dry particles are prepared, they can be separated, e.g., by filtration or cyclone centrifugation, to provide a particle sample having a preselected size distribution. For example, more than about 30%, more than about 40%, more than about 50%, more than about 60%, more than about 70%, more than about 80%, or more than about 90% of the respirable dry particles in the sample can have diameters within a predetermined range. The predetermined range within which a particular percentage of the respirable dry particles falls can be, for example, any of the size ranges described herein, e.g., a VMGD of about 0.1 to about 3 micrometers.

[0186] The suspension may be a nanosuspension as well as an intermediate for making a dry powder comprising the nanocrystalline compound.

[0187] The dry powder may be a drug embedded in a matrix material, such as sodium sulfate and leucine. Optionally, the dry powder may be spray dried to provide small, densely dispersed particles.

[0188] The dry powder can consist solely of the respirable dry particles described herein, without other carrier or excipient particles (also referred to as a "bulk powder"). If desired, the dry powder can include a blend of the respirable dry particles described herein with other carrier or excipient particles, such as lactose carrier particles that are greater than 10 micrometers, between 20 micrometers and 500 micrometers, and preferably between 25 micrometers and 250 micrometers. In some embodiments, dry powders that include carrier particles (blended powders) are excluded.

[0189] In a preferred embodiment, the dry powder does not include carrier particles. In one aspect, the angiogenesis inhibitor is embedded in a matrix including excipients and / or stabilizers. The dry powder may include a uniform content of respirable dry particles, each particle including the angiogenesis inhibitor. Thus, as used herein, "uniform content" means that all respirable particles include some amount of the angiogenesis inhibitor along with any stabilizers and / or excipients.

[0190] The dry powder can comprise respirable dry particles, wherein at least 98%, at least 99%, or substantially all (by weight) of the respirable dry particles comprise an angiogenesis inhibitor.

[0191] The dry powder can include an angiogenesis inhibitor distributed throughout a matrix containing one or more excipients. The excipients can include any number of salts, sugars, lipids, amino acids, surfactants, polymers, or other components suitable for pharmaceutical use. Preferred excipients can include sodium sulfate and leucine. The dry powder is typically produced by first processing an angiogenesis inhibitor (e.g., itraconazole, e.g., crystalline itraconazole) and adjusting the particle size using any number of techniques familiar to those skilled in the art (e.g., wet milling, jet milling). For example, the crystalline angiogenesis inhibitor can be processed with a stabilizer in an antisolvent to form a suspension. Preferred stabilizers include polysorbates (also known as TWEEN®), such as polysorbate 80 (PS80). Another preferred stabilizer is oleic acid or a salt thereof. The stabilized suspension of the crystalline angiogenesis inhibitor is then spray-dried with one or more additional excipients. The resulting dry particles contain the crystalline angiogenesis inhibitor dispersed throughout the excipient matrix, with each dry particle being homogeneous in composition.

[0192] In certain embodiments, the dry powders of the present invention are made by starting with a crystalline angiogenesis inhibitor (e.g., itraconazole), which can typically be obtained in the microcrystalline size range. The particle size of the microcrystalline angiogenesis inhibitor can be reduced to nanocrystalline size using any of a number of techniques familiar to those skilled in the art, including, but not limited to, high-pressure homogenization, high-shear homogenization, jet milling, pin milling, microfluidization, or wet milling (also known as ball milling, pearl milling, or bead milling). Wet milling is often preferred because it can achieve a wide range of particle size distributions, including those in the nanometer (<1 μm) size domain. Of particular importance in the submicron size domain is the use of surface-stabilizing components, such as surfactants (e.g., polysorbate 80, also known as TWEEN® 80). The surfactant allows for the creation of submicron particles during milling and the formation of a physically stable suspension. Surfactants isolate the many high-energy surfaces created during milling, preventing strong aggregation and precipitation. Therefore, the presence of surfactants is important for spray-drying homogeneous microparticles because they form a uniform and stable suspension, ensuring compositional uniformity throughout the particles. The use of surfactants allows for the formation of microsuspensions or nanosuspensions. The surfactant suspends nanocrystalline angiogenesis inhibitor (e.g., itraconazole) particles in a stable colloidal suspension in an antisolvent. The antisolvent for the drug can be water, or a combination of water and another miscible solvent, such as an alcohol or ketone, as a continuous antisolvent phase for the colloidal suspension. The feedstock to be spray-dried can be prepared by dissolving the soluble components in the desired solvent(s) and then dispersing the surfactant-stabilized crystalline angiogenesis inhibitor nanosuspension into the resulting feedstock with mixing, although the process is not limited to this specific order of operations.

[0193] Methods for analyzing the dry powders and / or respirable dry particles of the present invention are found in the Examples section below.

[0194] Liquid formulations Liquid formulations for delivery via pressurized metered dose inhalers (pMDIs) or soft mist inhalers (SMIs) can be prepared using any suitable method. For example, for use with a pMDI, the material may be prepared in a pressurized canister in which a crystalline microparticle form of an angiogenesis inhibitor (e.g., itraconazole) is suspended in a propellant, e.g., an HFA propellant or a CFC propellant, and optionally stabilized with a stabilizer, e.g., polysorbate 80. The pressurized suspension may then be delivered to the subject's respiratory tract by actuation of the pMDI. Table 1B includes various embodiments for delivering crystalline microparticle form of itraconazole via use of a pMDI. The nanoparticle solids concentration may vary from about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 50%. The dose volume of the pMDI may vary from about 20 μL to about 110 μL. The amount of itraconazole in the dosage volume may be about 15%, 20%, 25%, 30%, or 40%. The remainder of the volume may include a propellant and, optionally, a surfactant. The delivery efficiency of the pMDI may be about 15%, 20%, 25%, 30%, or 40%. The nominal dose of itraconazole in the pMDI may vary from about 0.50 mg to about 12 mg. For example, the nominal dose may be about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, or about 12 mg. The calculated delivered dose may range from about 0.1 mg to about 5 mg.

[0195] [Table 3]

[0196] For use with a soft mist inhaler (SMI), the angiogenesis inhibitor (e.g., itraconazole) is prepared in a solvent, e.g., water, in which it is poorly soluble, in crystalline microparticle form, and stabilized with a stabilizer, e.g., polysorbate 80. The suspension may be stored in a collapsible bag within a cartridge that is loaded into the device. A metered volume of the suspension is then passed through a capillary tube to a micropump. Upon activation of the SMI, a dose is delivered to the patient. Table 1C includes various embodiments for delivering itraconazole in crystalline microparticle form via the use of an SMI. The nanoparticle solids concentration may vary from about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, or about 50%. The SMI dosage volume may vary from about 10 μL to about 25 μL. The formulation may include itraconazole in crystalline microparticle form and a surfactant. The delivery efficiency of the SMI may be about 65%, 70%, 75%, 80%, or 85%. The nominal dose of itraconazole in the pMDI may vary from about 1.0 mg to about 8 mg. For example, the nominal dose may be about 2 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, or about 8 mg. The calculated delivered dose may range from about 0.5 mg to about 5 mg. [Table 4] [Example]

[0197] The materials used in the following examples and their sources are listed below. Sodium chloride, sodium sulfate, polysorbate 80, oleic acid, ammonium hydroxide, mannitol, magnesium lactate, and L-leucine were obtained from Sigma-Aldrich Co. (St. Louis, MO), Spectrum Chemicals (Gardena, CA), Applichem (Maryland Heights, MO), Alfa Aesar (Tewksbury, MA), Thermo Fisher (Waltham, MA), Croda Chemicals (East Yorkshire, United Kingdom), or Merck (Darmstadt, Germany). Itraconazole was obtained from Neuland (Princeton, NJ) or SMS Pharmaceutical Ltd. (Telengana State, India). Ultrapure water (ASTM Type II) was obtained from a water purification system (Millipore Corp., Billerica, MA) or equivalent.

[0198] method: The geometric or volume diameter of the suspension was determined using laser diffraction to determine the volume median diameter (×50 or Dv50), sometimes referred to as the volume geometric diameter (VMGD), of the active agent suspension. The apparatus consisted of a Horiba LA-950 instrument equipped with an automatic recirculation system or a fixed-volume sample cuvette for sample handling and removal. The sample was added to a dispersion medium consisting of either deionized water or deionized water containing less than 0.5% surfactant, such as polysorbate 80 or sodium dodecyl sulfate. Ultrasonic energy can be applied to aid in dispersing the suspension. When the laser transmittance was in the correct range, the sample was sonicated for 60 seconds at setting 5. The sample was then measured, and the particle size distribution was recorded.

[0199] Dry Powder Geometric or Volume Diameter Laser diffraction was used to determine the median volume diameter (×50 or Dv50), sometimes referred to as the median volume geometric diameter (VMGD), of the dry powders of the present invention. The apparatus consisted of a HELOS diffractometer and a RODOS dry powder disperser (Sympatec, Inc., Princeton, NJ). The RODOS disperser applies shear force to the particle sample and is controlled by the adjusted pressure of the compressed dry air blown through it (typically set at 1.0 bar at maximum orifice ring pressure). The pressure setting can be changed to vary the amount of energy used to disperse the powder. For example, the dispersion energy can be adjusted by changing the adjusted pressure from 0.2 bar to 4.0 bar. The powder sample is introduced into the RODOS funnel via a microspatula. The dispersed particles travel through a laser beam, and the resulting diffraction light pattern is collected by a series of detectors, typically using an R1 lens. The diffraction ensemble pattern was then converted to a volume-based particle size distribution using the Fraunhofer diffraction model, based on the fact that smaller particles diffract light at larger angles. Using this method, the span of the distribution was also determined according to the formula (Dv

[90] -Dv

[10] ) / (Dv

[50] ). The span value provides a relative measure of the polydispersity of the particle size distribution.

[0200] Aerodynamic Performance with an Andersen Cascade Impactor: The aerodynamic properties of powders dispersing from the inhalation device were evaluated using an Andersen Cascade Impactor (ACI) Mk-II 1 ACFM (Copley Scientific Limited, Nottingham, UK). The ACI device was operated under controlled environmental conditions of 18-25°C and 25-35% relative humidity (RH). The device consists of eight stages that separate aerosol particles based on inertial impaction. In each stage, the aerosol stream passes through a series of nozzles and impacts a corresponding impaction plate. Particles with sufficiently small inertia are carried forward into the aerosol stream, while the remaining particles impact the plate. In each subsequent stage, the aerosol passes through the nozzles at ever-increasing speeds, capturing increasingly smaller aerodynamic particles on the plate. After the aerosol passes through the final stage, a filter captures the smallest remaining particles (the so-called "final filter"). Gravimetric and / or chemical analysis can then be performed to determine the particle size distribution. A short-stack cascade impactor (also known as a collapsed cascade impactor) was also used to reduce the operating time required to assess two aerodynamic particle size cutpoints. This collapsed cascade impactor eliminated stages except those necessary to establish fine and coarse particle fractions. Depending on the impaction method used, two or eight separate powder fractions could be collected. Capsules (HPMC, No. 3, Capsugel Vcaps, Peapack, NJ) were filled with powder to a predetermined weight and placed into a handheld, breath-actuated dry powder inhaler (DPI) device: a high-resistance RS01™ DPI or an ultra-high-resistance UHR2 DPI (both manufactured by Plastiape, Osnago, Italy). The capsules were punctured, and the powder was drawn into the cascade impactor, which was operated for 2.0 seconds at a flow rate of 60.0 L / min.At this flow rate, the calibrated cutoff diameters are 8.6 micrometers, 6.5 micrometers, 4.4 micrometers, 3.3 micrometers, 2.0 micrometers, 1.1 micrometers, 0.5 micrometers, and 0.3 micrometers for the 8 stages, and 5.6 micrometers and 3.4 micrometers for the 2 stages used in the short stack cascade impactor, based on the Andersen cascade impactor. The fractions were collected by placing filters in the apparatus and determining the amount of powder that impacted the stages by gravitational or chemical measurement by HPLC.

[0201] Aerodynamic Performance with Next-Generation Impactor: The aerodynamic properties of powders dispersing from the inhalation device were evaluated using a Next-Generation Impactor (NGI) (Copley Scientific Limited, Nottingham, UK). For measurements using the NGI, the NGI device was operated under controlled environmental conditions of 18–25°C and 25–35% relative humidity (RH). The device consists of seven stages that separate aerosol particles based on inertial impaction and can operate at various airflow rates. In each stage, the aerosol stream passes through a series of nozzles and strikes a corresponding impaction surface. Particles with sufficiently small inertia are carried forward into the next stage, while the remaining particles strike this surface. In each subsequent stage, the aerosol passes through the nozzles at ever-increasing speeds, collecting increasingly aerodynamically smaller particles on its plate. After the aerosol leaves the final stage, a microorifice collector captures the smallest remaining particles. Gravimetric and / or chemical analysis can then be performed to determine the particle size distribution. Capsules (HPMC, No. 3, Capsugel Vcaps, Peapack, NJ) were filled with powder to a predetermined weight and placed into a handheld, breath-actuated dry powder inhaler (DPI) device: High Resistance RS01 DPI or Ultra High Resistance RS01 DPI (both manufactured by Plastiape, Osnago, Italy). The capsules were punctured, and the powder was drawn into a cascade impactor operated at a predetermined flow rate relative to 2.0 liters of inhaled air. The cutoff diameter of the stage at that flow rate was calculated. Wetted filters were placed in the device, and the amount of powder impacting the stages was determined by HPLC chemistry to capture these fractions.

[0202] Fine Particle Dose: The fine particle dose indicates the mass of one or more therapeutic agents in a given size range and can be used to predict the mass that will reach a specific region of the respiratory tract. Fine particle dose can be measured gravimetrically or chemically, either by ACI or NGI. When measured gravimetrically, the dry particles of the present invention are presumed to be homogeneous, and the mass of the therapeutic agent can be determined by multiplying the mass of powder in each stage and each collection filter by the percentage of the therapeutic agent in the formulation. When measured chemically, the powder in each stage or filter is collected, separated, and assayed, for example, by HPLC to determine the therapeutic agent content. The cumulative mass deposited on each stage at a given flow rate is calculated and the cumulative mass corresponding to a particle with a diameter of 5.0 micrometers is interpolated. This cumulative mass of powder contained in one or more capsules and acted upon in the impactor is equivalent to a fine particle dose of less than 5.0 micrometers (FPD<5.0 micrometers).

[0203] Mass Median Aerodynamic Diameter: Information obtained by the Andersen Cascade Impactor (ACI) was used to determine the mass median aerodynamic diameter (MMAD). The cumulative mass below the stage cutoff diameter was calculated for each stage and normalized by the amount of powder recovered. The MMAD for that powder was then formed by linear interpolation of the stage cutoff diameters, bracketing the 50th percentile. An alternative method for measuring MMAD is the Next Generation Impactor (NGI) method. Like the ACI, the MMAD is calculated by the cumulative mass below the stage cutoff diameter, bracketing the 50th percentile, for each stage and normalized by the amount of powder recovered. The MMAD for that powder was then formed by linear interpolation of the stage cutoff diameters, bracketing the 50th percentile.

[0204] Geometric or volumetric diameter of release: The median volume diameter (Dv50) (sometimes referred to as the median volume geometric diameter (VMGD)) of the powders of the present invention after release from a dry powder inhaler was determined using laser diffraction with a Spraytec (Malvern, Inc.) diffraction instrument. The powder was filled into size 3 capsules (V-Caps, Capsugel) and placed in a capsule-based dry powder inhaler (RS01™ Model 7 (high resistance), Plastiape, Italy), or DPI, which was then sealed in a cylinder. The cylinder was connected to a positive pressure air source, and the steady air flow through the system was measured with a mass flow meter and its duration was controlled by a timer-controlled solenoid valve. The outlet of the dry powder inhaler was exposed to room pressure, and the resulting aerosol jet was passed through the laser of a diffraction particle sizer (Spraytec) in its open bench configuration before being captured by a vacuum extractor. The steady air flow rate through the system was initiated using a solenoid valve. A steady airflow rate, typically 60 L / min, was inhaled through the DPI for a predetermined period, typically 2 seconds. Alternatively, airflow rates of 15, 20, or 30 L / min were inhaled through the DPI. The resulting geometric particle size distribution of the aerosol was calculated by software based on scattering pattern measurements from a photodetector, typically with samples taken at 1000 Hz during the inhalation period. The measured Dv50, GSD, and FPF < 5.0 μm were then averaged over the entire inhalation period.

[0205] The emitted dose (ED) refers to the mass of therapeutic agent that leaves a suitable inhalation device after exhalation or dispersion. The ED is determined using a method based on USP Section 601 Aerosols, Metered-Dose Inhalers and Dry Powder Inhalers, Delivered-Dose Uniformity, Sampling the Delivered Dose from Dry Powder Inhalers, United States Pharmacopeia convention, Rockville, MD, 13th Revision, pp. 222-225, 2007. The capsule contents are dispersed using either an RS01 HR inhaler with a pressure drop of 4 kPa and a typical flow rate of 60 LPM, or a UHR2 RS01 with a pressure drop of 4 kPa and a typical flow rate of 39 LPM. The emitted powder is collected on the filter of a filter holder sampler. The sampler is rinsed with an appropriate solvent, e.g., water, and analyzed using an HPLC method. In gravimetric analysis, a shorter filter holder sampling device is used to reduce the device's volume, and the filter is weighed before and after to determine the mass of powder delivered from the DPI to the filter. The therapeutic agent release amount is then calculated based on the content of the therapeutic agent in the delivered powder. The release amount can be recorded as the mass of therapeutic agent delivered from the DPI or as a percentage of the loading amount.

[0206] Thermogravimetric Analysis: Thermogravimetric analysis (TGA) was performed using either a Q500 model or a Discovery model thermogravimetric analyzer (TA Instruments, New Castle, DE). Samples were placed in either an open aluminum DSC pan or a closed aluminum DSC pan (which was then automatically pierced and opened prior to testing). The tare weight was pre-recorded by the instrument. The following method was used: a temperature ramp from ambient temperature (approximately 35°C) to 200°C at 5.00°C / min. Weight loss was recorded as a function of temperature up to 140°C. TGA allows the content of volatile compounds in the dry powders of the present invention to be calculated. When the process uses water alone or in combination with a volatile solvent, weight loss by TGA is a good estimate of water content.

[0207] Powder X-ray Diffraction: The crystalline nature of the formulations of the present invention was assessed by powder X-ray diffraction (PXRD). 20-30 mg material samples were analyzed on a powder X-ray diffractometer (D8 Discover with a LINXEYE detector (Bruker Corporation, Billerica, MA) or equivalent) using a 1.5418 A Cu X-ray tube with a scan range of 5-45° 2θ and a step size of 0.02° 2θ, with a data accumulation time of 1.2 seconds per step.

[0208] Itraconazole Content / Purity Using HPLC: A high-performance liquid chromatography (HPLC) method using a reversed-phase C18 column coupled to an ultraviolet (UV) detector has been developed for the analysis of the identity, bulk content, assay, CUPMD, and impurities of itraconazole dry powder. The reversed-phase column was equilibrated at 30°C, and the autosampler was set at 5°C. A gradient elution of the mobile phase, i.e., 20 mM monobasic sodium phosphate (mobile phase A) at pH 2.0 and acetonitrile (mobile phase B), was used, ranging from 59:41 (A:B) to 5:95 (A:B) over a run time of 19.5 minutes. Detection was by UV at 258 nm, with an input volume of 10 μL. The itraconazole content in the powder of the present invention was quantified against a standard curve.

[0209] The identification of known impurities A, B, C, D, E, F, and G (as identified in monograph Ph.Eur.01 / 2011:1335) is confirmed by comparing the retention times of the impurity peaks in the itraconazole dry powder sample with those of the reference standard itraconazole USP impurity mixture spiked with impurity A. Unknown impurities are identified and quantified by their retention times relative to the retention time of the main itraconazole peak, along with the area above the limit of detection (LOD). All impurities are measured by their area percentage relative to the itraconazole peak.

[0210] Particle size reduction: The particle size distribution of the crystalline active agents of the present invention can be controlled using a number of techniques familiar to those skilled in the art, including, but not limited to, high-pressure homogenization, high-shear homogenization, jet milling, pin milling, microfluidization, or wet milling (also known as ball milling, pearl milling, or bead milling). Wet milling is often preferred because it can achieve a wide range of particle size distributions, including those in the nanometer (less than 1 μm) size domain.

[0211] Particle size reduction using low-energy wet milling: One technique for reducing the particle size of the active agents of the present invention was by low-energy wet milling (also known as roller milling or jar milling). A suspension of the active agent was prepared in a poor solvent, which could be water or any solvent in which the active agent is poorly soluble. A stabilizer (which can be, but is not limited to, a nonionic surfactant or an amphiphilic polymer) can then be added to the suspension along with grinding media (which can be, but is not limited to, spheres with high abrasion resistance and a size range of 0.03 to 0.70 millimeters in diameter). The suspension is then rotated using a jar mill (US Stoneware, East Palestine, OH, USA), with samples taken periodically to assess particle size (LA-950, HORIBA, Kyoto, Japan). Once the particle size is sufficiently reduced or the minimum particle size is reached, the suspension is sieved to remove the grinding media and the product is recovered.

[0212] Particle size reduction using high-energy wet milling: Another technique for reducing the particle size of the active agents of the present invention was by high-energy wet milling using a rotor-stator mill or a media-agitated mill. A suspension of the active agent was prepared in a poor solvent, which could be water or any solvent in which the active agent is poorly soluble. A stabilizer (which can be, but is not limited to, a nonionic surfactant or an amphiphilic polymer) can then be added to the suspension along with grinding media (which can be, but is not limited to, spheres with high abrasion resistance and a size range of 0.03 to 0.70 millimeters in diameter). The suspension is then fed into the mill, which can operate in either batch or recirculation mode. The process consists of the suspension and grinding media being agitated in a milling chamber, which increases the amount of energy input to the system and accelerates the particle size reduction process. The milling chamber and recirculation vessel are jacketed and actively cooled to avoid product temperature rise. The agitation and recirculation rates of the suspension are controlled during the process. Samples are taken periodically to evaluate particle size (LA-950, HORIBA, Kyoto, Japan). Once the particle size is sufficiently reduced or the minimum particle size is reached, the suspension is discharged from the mill.

[0213] Particle size reduction using microfluidization: Another technique for narrowing the particle size distribution of the active agents of the present invention was via microfluidization. Microfluidizer-based processing is the operation of a high-shear wet processing unit utilized for particle size reduction of liquids and solids. The unit can consist of various interaction chambers, which are cylindrical modules with predetermined orifice and channel designs through which fluids pass at high pressure to control shear rates. Product enters the unit via an inlet vessel and is forced into a geometrically defined interaction chamber by a high-pressure pump at velocities of up to 400 m / s. The product is then efficiently cooled, if necessary, and collected in an output vessel. This process can be repeated (e.g., multiple "passes") as needed to achieve particle size targets. The particle size of the active agents of the present invention is periodically monitored by laser diffraction (LA-950, HORIBA, Kyoto, Japan). Once the particle size is sufficiently reduced or the minimum particle size is reached, the suspension is withdrawn from the unit.

[0214] Particle size reduction using jet milling: Another technique for narrowing the particle size distribution of the active agents of the present invention has been through jet milling. Jet mills use fluid energy (compressed air or gas) to crush and classify in a single chamber with no moving parts. The particles are driven by high-pressure air and accelerated into a shallow crushing chamber at high speeds. As the particles collide with each other, their size is reduced. Centrifugal force holds the larger particles in the rotating crushing zone until they achieve the desired fine particle size. Centripetal force drives the desired particles toward an electrostatic classifier where they can be discharged once they have achieved the correct particle size. The final particle size is controlled by varying the feed rate and propellant pressure.

[0215] Preparation of Liquid Feedstocks for Spray Drying Spray drying homogeneous particles requires that the components of interest be solubilized in a solution or suspended in a uniform and stable suspension. The feedstock can utilize water, or a combination of water and other miscible solvents, such as alcohols or ketones, as the solvent for solutions or the continuous phase for suspensions. Various formulation feedstocks were prepared by dissolving the soluble components in the desired solvent(s) and then dispersing a surfactant-stabilized active agent-containing suspension into the resulting solution with mixing, although the process is not limited to this given sequence of operations.

[0216] Spray drying using a Niro spray dryer: Dry powder was produced by spray drying using a Niro Mobile Minor spray dryer (GEA Process Engineering Inc., Columbia, MD), with the powder collected from a cyclone, a product filter, or both. Atomization of the feed liquid was performed using a co-current two-fluid nozzle, either a Niro (GEA Process Engineering Inc., Columbia, MD) or a Spraying Systems (Carol Stream, IL) two-fluid nozzle 1 / 4J, with a gas cap 671471 and a fluid cap 2850SS, although other two-fluid nozzle configurations are possible. In some embodiments, the two-fluid nozzle can be configured for internal or external mixing. Additional atomization techniques include rotary atomization or pressure nozzles. The feed liquid was fed into the two-fluid nozzle using a gear pump (Cole-Parmer Instrument Company, Vernon Hills, IL) directly or via a static mixer (Charles Ross & Son Company, Hauppauge, NY) just prior to introduction into the two-fluid nozzle. Additional liquid delivery techniques include delivery from a pressurized vessel. Nitrogen or air may be used as the drying gas, provided that the moisture in the air is at least partially removed before use. Pressurized nitrogen or air can be used as the atomizing gas supplied to the two-fluid nozzle. The drying gas inlet temperature can range from 70°C to 300°C, the outlet temperature can range from 30°C to 120°C, and the liquid feed rate can range from 10 mL / min to 100 mL / min. The gas supplied to the two-fluid atomizer can vary depending on the nozzle selection, ranging from 5 kg / hr to 50 kg / hr for the Niro co-current two-fluid nozzle and from 30 g / min to 150 g / min for the Spraying Systems 1 / 4J two-fluid nozzle. The atomizing gas rate can be set to achieve a specific gas-to-liquid mass ratio, which directly affects the droplet size produced. The pressure in the drying drum can range from +3°C to -6°C.The spray-dried powder can be collected in a vessel, at the outlet of the cyclone, on a cartridge or baghouse filter, or from both the cyclone and the cartridge or baghouse filter.

[0217] Spray drying using a Buchi spray dryer: Dry powders were prepared by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), and the powder was collected from either a standard cyclone or a high-performance cyclone. The system was operated in open-loop (single-pass) mode with either air or nitrogen as the drying and atomizing gas. When operating with air, the system used a Buchi B-296 dehumidifier to ensure stable temperature and humidity of the air used for spray drying. Additionally, an external LG dehumidifier (model 49007903, LG Electronics, Englewood Cliffs, NJ) was operated continuously when the room relative humidity exceeded 30% RH. When operating with nitrogen, a pressurized nitrogen source was used. Additionally, the system aspirator was adjusted to maintain a system pressure of -2.0 inches of water. Atomization of the feed liquid was performed using a 1.5 mm diameter Buchi two-fluid nozzle or a Schlick 970-0 atomizer (Dusen-Schlick GmbH, Coburg, Germany) equipped with a 0.5 mm liquid insert. The inlet temperature of the process gas ranged from 100°C to 220°C, the outlet temperature from 30°C to 120°C, and the liquid feed flow rate from 3 mL / min to 10 mL / min. The two-fluid atomizing gas was 25 mm to 45 mm (300 LPH to 530 LPH) for the Buchi two-fluid nozzle, and the upper atomizing air pressure for the Schlick atomizer was 0.3 bar. The aspirator speed ranged from 50% to 100%.

[0218] Stability Assessment: As detailed in the International Conference on Harmonization (ICH) Q1 guidance, the physicochemical stability and aerosol performance of a given formulation were evaluated at 2-8°C, 25°C / 60% RH, and 40°C / 75% RH when material availability permitted. Stability samples were stored in calibrated chambers (Models PH024 and PH074 from Darwin Chambers Company, St. Louis, MO). Bulk powder samples were weighed into amber glass vials, sealed at 30% RH, and induction-sealed in aluminum pouches (Drishield 3000, 3M, St. Paul, MN) with silica desiccant (2.0 g, Multisorb Technologies, Buffalo, NY). Additionally, to assess the stability of the formulation in capsules, target masses of powder were manually weighed into No. 3 HPMC capsules (Capsugel Vcaps, Peapack, NJ) with a tolerance of ±0.2 mg at 30% RH. The filled capsules were then divided into high density polyethylene (HDPE) bottles and sealed in aluminum pouches with silica desiccant by induction sealing.

[0219] Example 1. Dry Powder Formulation of Nanocrystalline Itraconazole Stabilized with Polysorbate 80 and Containing Sodium Sulfate / Mannitol A. Powder preparation Nanocrystalline itraconazole was prepared by combining 11.662 g of itraconazole (Neuland lot ITI0114005) with 103.789 g of water and 1.1662 g of polysorbate 80 (Spectrum lot 2DI0112). Subsequently, 129.625 g of 500 μm grinding media polystyrene (Dow Chemical, Midland MI) was added to the suspension, and the suspension was milled at 1000 rpm for 1 hour, then at 1500 rpm for 30 minutes before collection. The final median particle size (Dv(50)) of the milled suspension was 124 nm.

[0220] A feedstock solution was prepared and used to manufacture dry powders composed of nanocrystalline itraconazole, polysorbate 80, and other additional excipients. Drug loadings of 20 wt% and 50 wt% itraconazole on a dry basis were targeted. The feedstock solution used to spray-dry the particles was made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and stirred until the solution was visually clear. The itraconazole-containing suspension was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock was stirred while spray-drying. The feedstock mass was 83.3 g, which supported a 15-minute manufacturing campaign. Table 2 lists the components of the feedstock used to prepare the dry powder. [Table 5]

[0221] Dry powders of Formulations I and II were produced from these raw materials by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), with the powder collected in a cyclone. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. A Buchi nozzle with a 1.5 mm cap and 0.7 liquid tip was used for atomization of the feed liquid. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0222] The dry powders were produced according to the following spray-drying conditions: For Formulations I and II, the solids concentration of the liquid feed was 3.0 wt%, the inlet temperature of the process gas was 117-119°C, the outlet temperature of the process gas was 50°C, the flow rate of the drying gas was 17.0 kg / hr, the flow rate of the atomizing gas was 30.4 g / min, and the flow rates of the atomizing gas and liquid feed were 6.0 mL / min. The resulting dry powders are recorded in Table 3 below. [Table 6]

[0223] B. Powder Characterization The bulk particle size characteristics of the two formulations can be seen in Table 4. The span at 1 bar of 1.83 for Formulation I and 1.67 for Formulation II indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersibility ratio of 1.07 for Formulation I and 1.12 for Formulation II indicates that the formulations are relatively insensitive to dispersion energy, a desirable feature that allows for similar particle dispersion over a range of dispersion energies. [Table 7]

[0224] The measured and / or calculated geometric particle size and capsule emitted powder mass (CEPM) simulated patient flow rates at 60 liters per minute (LPM) and 20 LPM were measured for the two formulations and are recorded in Table 5. The small changes in CEPM and geometric size from 60 LPM to 20 LPM indicate that the dry powder is relatively unaffected by patient inspiratory flow, resulting in relatively similar therapeutic doses delivered to patients breathing at various flow rates. [Table 8]

[0225] The aerodynamic particle size, fraction of fine particles, and fine particle dose measured and / or calculated in an 8-stage Andersen cascade impactor (ACI-8) are recorded in Table 6. The fine particle doses for both Formulations I and II indicate that a high percentage of the nominal dose loaded into the capsule reaches the impactor stage (38.8% for Formulation I and 37.1% for Formulation II) and is therefore predicted to be delivered to the lung. The MMAD for Formulation I was 3.59 micrometers and the MMAD for Formulation II was 3.17 micrometers, indicating deposition in the central and conducting airways. [Table 9]

[0226] The weight loss of Formulations I and II was determined by TGA and was found to be 0.48% and 0.15%, respectively.

[0227] The itraconazole contents of Formulations I and II were determined by HPLC-UV to be 102.9% and 103.1%, respectively.

[0228] The crystallinity of Formulations I and II was assessed by XRD. Diffraction patterns of itraconazole were observed for both formulations, suggesting that the milling or spray-drying process did not affect the solid state of itraconazole. Additional peaks observed in the patterns correspond to additional excipients in the formulations (Figure 1).

[0229] Formulations I and II were determined to be stable after storage at 2-8°C and 25°C / 60% RH for 6 months.

[0230] Example 2. Dry Powder Formulation of Nanocrystalline Itraconazole Stabilized with Polysorbate 80 and Containing Sodium Chloride / Leucine A. Powder preparation Nanocrystalline itraconazole was prepared by combining 11.662 g of itraconazole (Neuland lot ITI0114005) with 103.789 g of water and 1.1662 g of polysorbate 80 (Spectrum lot 2DI0112). Subsequently, 129.625 g of 500 μm grinding media polystyrene (Dow Chemical, Midland MI) was added to the suspension, and the suspension was milled at 1000 rpm for 1 hour, then at 1500 rpm for 30 minutes before collection. The final median particle size (Dv(50)) of the milled suspension was 124 nm.

[0231] A feedstock solution was prepared and used to manufacture dry powders composed of nanocrystalline itraconazole, polysorbate 80, and other additional excipients. Drug loadings of 20 wt% and 50 wt% itraconazole on a dry basis were targeted. The feedstock solution used to spray-dry the particles was made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and stirred until the solution was visually clear. The itraconazole-containing suspension was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock was stirred while spray-drying. The feedstock mass was 83.3 g, which supported a 15-minute manufacturing campaign. Table 7 lists the components of the feedstock used to prepare the dry powder. [Table 10]

[0232] Dry powders of Formulations I and II were produced from these raw materials by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), with the powder collected in a cyclone. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. A Buchi nozzle with a 1.5 mm cap and 0.7 liquid tip was used for atomization of the feed liquid. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0233] The dry powders were produced according to the following spray-drying conditions: For Formulations I and II, the solids concentration of the liquid feed was 3.0%, the inlet temperature of the process gas was 138°C-141°C, the outlet temperature of the process gas was 60°C, the flow rate of the drying gas was 17.0 kg / hr, the flow rate of the atomizing gas was 30.4 g / min, and the flow rates of the atomizing gas and liquid feed were 6.0 mL / min. The resulting dry powders are recorded in Table 8 below. [Table 11]

[0234] B. Powder Characterization The bulk particle size characteristics of these two formulations can be seen in Table 9. The span at 1 bar of 1.76 for Formulation III and 1.86 for Formulation IV indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersibility ratio of 1.19 for Formulation III and 1.05 for Formulation IV indicates that the formulations are relatively insensitive to dispersion energy, a desirable feature that allows for similar particle dispersion over a range of dispersion energies. [Table 12]

[0235] The measured and / or calculated geometric particle size and capsule emitted powder mass (CEPM) simulated patient flow rates at 60 liters per minute (LPM) and 20 LPM were measured for these two formulations and are recorded in Table 10. The small changes in CEPM and geometric size from 60 LPM to 20 LPM indicate that the dry powder is relatively unaffected by patient inspiratory flow, resulting in relatively similar therapeutic doses delivered to patients breathing at various flow rates. [Table 13]

[0236] The aerodynamic particle size, fraction of fine particles, and fine particle dose measured and / or calculated in an 8-stage Andersen cascade impactor (ACI-8) are recorded in Table 11. The fine particle doses for both Formulations III and IV indicate that a high percentage of the nominal dose loaded into the capsule reaches the impactor stage (55.5% for Formulation III and 49.4% for Formulation IV) and is therefore predicted to be delivered to the lung. The MMAD for Formulation III was 3.14 micrometers and the MMAD for Formulation IV was 3.30 micrometers, indicating deposition in the central and conducting airways. [Table 14]

[0237] The weight loss of Formulations III and IV was determined by TGA and was found to be 0.15% and 0.08%, respectively.

[0238] The itraconazole contents of Formulations III and IV were determined by HPLC-UV to be 103.7% and 104.9%, respectively.

[0239] The crystallinity of Formulations III and IV was assessed by XRD. Diffraction patterns of itraconazole were observed for both formulations, suggesting that the milling or spray-drying process did not affect the solid state of itraconazole. Additional peaks observed in the patterns correspond to additional excipients in the formulations (Figure 2).

[0240] Formulations III and IV were determined to be stable after storage at 2-8°C and 25°C / 60% RH for 6 months.

[0241] Example 3. Dry Powder Formulation of Nanocrystalline Itraconazole Stabilized with Polysorbate 80 and Containing Magnesium Lactate / Leucine A. Powder preparation Nanocrystalline itraconazole was prepared by combining 11.662 g of itraconazole (Neuland lot ITI0114005) with 103.789 g of water and 1.1662 g of polysorbate 80 (Spectrum lot 2DI0112). Subsequently, 129.625 g of 500 μm grinding media polystyrene (Dow Chemical, Midland MI) was added to the suspension, and the suspension was milled at 1000 rpm for 1 hour, then at 1500 rpm for 30 minutes before collection. The final median particle size (Dv(50)) of the milled suspension was 124 nm.

[0242] A feedstock solution was prepared and used to manufacture dry powders composed of nanocrystalline itraconazole, polysorbate 80, and other additional excipients. Drug loadings of 20 wt% and 50 wt% itraconazole on a dry basis were targeted. The feedstock solution used to spray-dry the particles was made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and stirred until the solution was visually clear. The itraconazole-containing suspension was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock was stirred while spray-drying. The feedstock mass was 83.3 g, which supported a 15-minute manufacturing campaign. Table 12 lists the components of the feedstock used to prepare the dry powder. [Table 15]

[0243] Dry powders of Formulations V and VI were produced from these raw materials by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), with the powder collected in a cyclone. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. A Buchi nozzle with a 1.5 mm cap and a 0.7 liquid tip was used for atomization of the feed liquid. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0244] The dry powders were produced according to the following spray-drying conditions: For Formulations V and VI, the solids concentration of the liquid feed was 3.0%, the inlet temperature of the process gas was 171-173°C, the outlet temperature of the process gas was 80°C, the flow rate of the drying gas was 17.0 kg / hr, the flow rate of the atomizing gas was 30.4 g / min, and the flow rates of the atomizing gas and liquid feed were 6.0 mL / min. The resulting dry powders are recorded in Table 13 below. [Table 16]

[0245] B. Powder Characterization The bulk particle size characteristics of these two formulations can be seen in Table 14. The span at 1 bar of 1.70 for Formulation V and 1.83 for Formulation VI indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersibility ratio of 1.02 for Formulation V and 1.05 for Formulation VI indicates that the formulations are relatively insensitive to dispersion energy, a desirable feature that allows for similar particle dispersion over a range of dispersion energies. [Table 17]

[0246] The measured and / or calculated geometric particle size and capsule emitted powder mass (CEPM) simulated patient flow rates at 60 liters per minute (LPM) and 20 LPM were measured for these two formulations and are recorded in Table 15. The small changes in CEPM and geometric size from 60 LPM to 20 LPM indicate that the dry powder is relatively unaffected by patient inspiratory flow, resulting in relatively similar therapeutic doses delivered to patients breathing at various flow rates. [Table 18]

[0247] The aerodynamic particle size, fraction of fine particles, and fine particle dose measured and / or calculated in an 8-stage Andersen cascade impactor (ACI-8) are recorded in Table 16. The fine particle doses of both Formulations V and VI indicate that a high percentage of the nominal dose loaded into the capsule reaches the impactor stage (39.6% for Formulation V and 44.6% for Formulation VI) and is therefore predicted to be delivered to the lung. The MMAD for Formulation V was 3.97 micrometers and the MMAD for Formulation VI was 3.42 micrometers, indicating deposition in the central and conducting airways. [Table 19]

[0248] The weight loss of Formulations V and VI was determined by TGA and was found to be 5.157% and 3.087%, respectively.

[0249] The itraconazole content of Formulations V and VI was determined by HPLC-UV to be 99.7% and 100.6%, respectively.

[0250] The crystallinity of Formulations V and VI was assessed by XRD. Diffraction patterns of itraconazole were observed for both formulations, suggesting that the milling or spray-drying process did not affect the solid state of itraconazole. Additional peaks observed in the patterns correspond to additional excipients in the formulations (Figure 3).

[0251] Formulations V and VI were determined to be stable after storage for 6 months at 2-8°C and 25°C / 60% RH.

[0252] Example 4. Dry Powder Formulation of Oleic Acid Stabilized Nanocrystalline Itraconazole with Sodium Sulfate / Leucine A. Powder preparation Nanocrystalline itraconazole was prepared by combining 11.646 g of itraconazole (Neuland lot ITI0114005) with 104.233 g of water, 0.582 g of oleic acid (Croda000705097), and 9.44 g of 10% ammonium hydroxide. Subsequently, 129.625 g of 500 μm grinding media polystyrene (Dow Chemical, Midland MI) was added to the suspension, and the suspension was milled at 1000 rpm for 1 hour, then at 1500 rpm for an additional hour before collection. The final median particle size (Dv(50)) of the milled suspension was 120 nm.

[0253] A feedstock solution was prepared and used to manufacture dry powders composed of nanocrystalline itraconazole, oleic acid, and other additional excipients. Drug loadings of 50 wt% and 70 wt% itraconazole on a dry basis were targeted. The feedstock solutions used to spray-dry the particles were made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and stirred until the solution was visually clear. The itraconazole-containing suspension was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock was stirred while spray-drying. Feedstock amounts ranged from 100 to 193.3 g, which supported campaigns lasting from 16 to 34 minutes. Table 17 lists the components of the feedstock used to prepare the dry powders. [Table 20]

[0254] Dry powders of Formulations VII and VIII were produced from these raw materials by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), with the powder collected in a cyclone. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. A Buchi nozzle with a 1.5 mm cap and 0.7 liquid tip was used for atomization of the feed liquid. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0255] The dry powders were produced according to the following spray-drying conditions: For Formulations VII and VIII, the solids concentration of the liquid feed was 3.0%, the inlet temperature of the process gas was 131°C-133°C, the outlet temperature of the process gas was 60°C, the drying gas flow rate was 17.0 kg / hr, the atomizing gas flow rate was 30.4 g / min (1.824 kg / hr), and the liquid feed flow rate was 6.0 mL / min. The resulting dry powders are recorded in Table 18 below. [Table 21]

[0256] B. Powder Characterization The bulk particle size characteristics of these two formulations can be seen in Table 19. The span at 1 bar of 1.94 for Formulation VII and 1.81 for Formulation VIII indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersibility ratio of 1.22 for Formulation VII and 1.11 for Formulation VIII indicates that the formulations are relatively insensitive to dispersion energy, a desirable feature that allows for similar particle dispersion over a range of dispersion energies. [Table 22]

[0257] The measured and / or calculated geometric particle size and capsule emitted powder mass (CEPM) simulated patient flow rates at 60 liters per minute (LPM) and 20 LPM were measured for these two formulations and are recorded in Table 20. The small changes in CEPM and geometric size from 60 LPM to 20 LPM indicate that the dry powder is relatively unaffected by patient inspiratory flow, resulting in relatively similar therapeutic doses delivered to patients breathing at various flow rates. [Table 23]

[0258] The aerodynamic particle size, percent fine particle size, and fine particle dose measured and / or calculated in an 8-stage Andersen cascade impactor (ACI-8) are recorded in Table 21. The fine particle doses of both Formulations VII and VIII indicate that a high percentage of the nominal dose loaded into the capsule reaches the impactor stage (56.0% for Formulation VII and 52.6% for Formulation VIII) and is therefore predicted to be delivered to the lung. The MMAD for Formulation VII was 2.77 micrometers and the MMAD for Formulation VIII was 3.08 micrometers, indicating deposition in the central and conducting airways. [Table 24]

[0259] The weight loss of Formulations VII and VIII was determined by TGA and was found to be 0.47% and 0.33%, respectively.

[0260] The itraconazole content of Formulations VII and VIII was determined by HPLC-UV to be 101.5% and 101.4%, respectively.

[0261] The crystallinity of Formulations VII and VIII was assessed by XRD. Diffraction patterns of itraconazole were observed for both formulations, suggesting that the milling or spray-drying process did not affect the solid state of itraconazole. Additional peaks observed in the patterns correspond to additional excipients in the formulations (Figure 4).

[0262] Formulations VII and VIII were determined to be stable after storage at 2-8°C and 25°C / 60% RH for 6 months.

[0263] Example 5. Reference Liquid Nanocrystalline and Microcrystalline Itraconazole Formulations A liquid formulation of crystalline particulate itraconazole was prepared.

[0264] Formulation IX is a microsuspension of itraconazole with polysorbate 80. The itraconazole concentration in the liquid is 5 mg / mL. The ratio of itraconazole to polysorbate 80 is 10:1 (wgt / wgt). The median size of the itraconazole crystals is 1600 nanometers.

[0265] Formulation X is a nanosuspension of itraconazole with polysorbate 80. The itraconazole concentration in the liquid is 5 mg / mL. The ratio of itraconazole to polysorbate 80 is 10:1 (wgt / wgt). The median size of the itraconazole crystals is 132 nanometers.

[0266] Example 6. Dry Powder Formulation of Oleic Acid Stabilized Nanocrystalline Itraconazole with Sodium Sulfate / Leucine A. Powder preparation Nanocrystalline itraconazole was prepared by combining 30.374 g of itraconazole (Neuland ITI0714011) with 87.018 g of water, 1.519 g of oleic acid (Croda 000705097), and 2.585 g of ammonium hydroxide (Acros B0522464). Subsequently, 129.625 g of 500 μm grinding media polystyrene (Dow Chemical, Midland MI) was added to the suspension, and the suspension was milled at 1800 rpm for 2 hours before collection. The final median particle size (Dv(50)) of the milled suspension was 124 nm.

[0267] A feedstock solution was prepared and used to manufacture a dry powder composed of nanocrystalline itraconazole, oleic acid, and other additional excipients. A drug loading of 50 wt% itraconazole on a dry basis was targeted. The feedstock solution used to spray-dry the particles was made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and stirred until the solution was visually clear. The itraconazole-containing suspension was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock mass was 1219.4 g, which supported an approximately 3.5-hour manufacturing campaign. Table 22 lists the components of the feedstock used to prepare the dry powder. [Table 25]

[0268] Dry powder of Formulation XI was produced from this raw material by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), with the powder collected in a cyclone. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. A Buchi nozzle with a 1.5 mm cap and 0.7 liquid tip was used for atomization of the feed liquid. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0269] The dry powders were produced according to the following spray-drying conditions: For Formulation XI, the solids concentration of the liquid feedstock was 3.0%, the inlet temperature of the process gas was 129°C-132°C, the outlet temperature of the process gas was 60°C, the drying gas flow rate was 17.0 kg / hr, the atomizing gas flow rate was 30.4 g / min, and the liquid feedstock flow rate was 6.0 mL / min. The resulting dry powders are recorded in Table 23 below. [Table 26]

[0270] B. Powder Characterization The bulk particle size characteristics of the formulations are found in Table 24. The span at 1 bar of 2.77 for Formulation XI indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersibility ratio of 1.28 for Formulation XI indicates that the particle size is relatively insensitive to dispersion energy, a desirable feature that allows for similar dispersion over a range of dispersion energies. [Table 27]

[0271] The measured and / or calculated geometric particle size and simulated patient flow rate of capsule emitted powder mass (CEPM) at 60 liters per minute (LPM) and 20 LPM were measured for this formulation and are recorded in Table 25. The small changes in CEPM and geometric size from 60 LPM to 20 LPM indicate that the dry powder is relatively unaffected by patient inspiratory flow, resulting in relatively similar therapeutic doses delivered to patients breathing at various flow rates. [Table 28]

[0272] The aerodynamic particle size, fraction of fine particles, and fine particle dose measured and / or calculated by the Next Generation Impactor (NGI) are recorded in Table 26. The fine particle dose of Formulation XI indicates that a high percentage of the nominal dose loaded into the capsule reaches the impactor stage (42%) and is therefore predicted to be delivered to the lung. The MMAD of Formulation XI was 3.37 micrometers, indicating deposition in the central and conducting airways. [Table 29]

[0273] The weight loss of Formulation XI was determined by TGA and was found to be 0.31%. The itraconazole content of Formulation XI was determined by HPLC-UV to be 99.7%. The crystallinity of Formulation XI was assessed by XRD. The diffraction pattern of itraconazole was observed in the formulation, suggesting that the milling or spray drying process did not affect the solid state of itraconazole. Additional peaks observed in the pattern correspond to additional excipients in the formulation (Figure 5).

[0274] Example 7. Dry Powder Formulations of Nanocrystalline Itraconazole of Various Particle Sizes Stabilized with Polysorbate 80 and Containing Sodium Sulfate / Leucine A. Powder preparation Formulation XII nanocrystalline itraconazole was prepared by blending 30.090 g of itraconazole (Neuland ITI0114005 and ITI0714011) with 87.262 g of water and 3.009 g of polysorbate 80. Subsequently, 129.625 g of 500 μm grinding media polystyrene (Dow Chemical, Midland MI) was added to the suspension, and the suspension was milled at 1800 rpm for 1 hour before collection. The final median particle size (Dv(50)) of the milled suspension was 132 nm. This process is hereinafter referred to as "Wet Mill Process #1."

[0275] Formulation XIII, nanocrystalline itraconazole, was prepared as a suspension containing 10 wt% itraconazole and 1.0 wt% polysorbate 80 in deionized water. The polysorbate 80 was dissolved in 89.0% DI water using a magnetic stir bar, and then the itraconazole was slowly added, also using a magnetic stir bar. Once all the itraconazole was suspended, the formulation was processed in a Microfluidizer processor M-110P at 30,000 psi for 120 passes, using an ice-water cooling coil to cool the material during processing. The final median particle size (Dv(50)) of the milled suspension was 198 nm. This process is hereafter referred to as "Microfluidics Process #1."

[0276] Formulation XIV of nanocrystalline itraconazole was prepared by blending 30.090 g of itraconazole (Neuland ITI0114005) with 87.26195 g of water and 3.009 g of polysorbate 80. Subsequently, 129.625 g of 500 μm grinding media polystyrene (Dow Chemical, Midland MI) was added to the suspension, and the suspension was milled at 1000 rpm for 30 minutes before collection. The final median particle size (Dv(50)) of the milled suspension was 258 nm. This process is hereinafter referred to as "Wet Mill Process #2."

[0277] Formulation XV, microcrystalline itraconazole, was prepared using a jet mill Qualification Micronizer (Sturtevant, Hanover, MA, USA). The feed pressure was set at 90 psig, and the milling pressure was set at 40 psig. Itraconazole was continuously fed into the mill until 60.3 g of itraconazole was milled. The final median particle size (Dv(50)) of the milled API was 1600 nm. This process is hereafter referred to as "Jet Mill Process #1." Formulation XV, micronized itraconazole, was then blended into a suspension consisting of 10 wt% itraconazole and 1.0 wt% polysorbate 80 in deionized water. The batch size was 200 g. The polysorbate 80 was dissolved in 89.0% DI water using a magnetic stir bar, and then the itraconazole was slowly added and mixed until the suspension was visually dispersed and uniform.

[0278] A feedstock solution was prepared and used to manufacture a dry powder composed of crystalline itraconazole, polysorbate 80, and other additional excipients. A drug loading of 50 wt% itraconazole on a dry basis was targeted. The feedstock solution used to spray-dry the particles was made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and stirred until the solution was visually clear. The itraconazole-containing suspension was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock was stirred while spray-drying. The feedstock mass ranged from 166.67 g to 1219.4 g, and this mass supported a 30-minute to 3.5-hour manufacturing campaign. Table 27 lists the components of the feedstock used to prepare the dry powder. [Table 30]

[0279] Dry powders of Formulations XII-XV were produced from these raw materials by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), with the powder collected in a cyclone. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. A Buchi nozzle with a 1.5 mm cap and 0.7 liquid tip was used for atomization of the feed liquid. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0280] The dry powders were produced according to the following spray-drying conditions: For Formulations XII, XIV, and XV, the solids concentration of the liquid feed was 3%, the inlet temperature of the process gas was 127°C-140°C, the outlet temperature of the process gas was 60°C, the flow rate of the drying gas was 17.0 kg / hr, the flow rate of the atomizing gas was 30.0 g / min, and the flow rate of the liquid feed was 6.0 mL / min. The resulting dry powders are recorded in Table 28 below.

[0281] The dry powders were produced according to the following spray drying conditions: For Formulation XIII, the solids concentration of the liquid feed was 3%, the inlet temperature of the process gas was 134°C, the outlet temperature of the process gas was 60°C, the flow rate of the drying gas was 17.0 kg / hr, the flow rate of the atomizing gas was 30.4 / min, and the flow rate of the liquid feed was 6.0 mL / min. The resulting dry powders are recorded in Table 28 below. [Table 31]

[0282] B. Powder Characterization The bulk particle size properties of the four formulations can be seen in Table 29. The span at 1 bar of less than 2.10 for formulations XII-XV indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersion ratio of 1.25 for formulations XII-XV indicates that they are relatively independent of dispersion energy, a desirable property that allows for similar particle dispersion across a range of dispersion energies. [Table 32]

[0283] Measured and / or calculated geometric particle size and capsule emitted powder mass (CEPM) simulated patient flow rates at 60 liters per minute (LPM) and 30 LPM were measured for Formulations XII, XIV, and XV and are recorded in Table 30. The small changes in CEPM and geometric size from 60 LPM to 30 LPM indicate that the dry powder is relatively unaffected by patient inspiratory flow, resulting in relatively similar therapeutic doses delivered to patients breathing at various flow rates. Formulation XIII was not tested for emitted particle size due to the lack of sufficient material. [Table 33]

[0284] The aerodynamic particle size, fraction of fine particles, and fine particle dose measured and / or calculated using an 8-stage Andersen Cascade Impactor (ACI-8) or Next Generation Impactor (NGI) are recorded in Table 31. The fine particle doses of all Formulations XII-XV demonstrated that greater than 30% of the nominal dose reached the impactor stage and would therefore be predicted to be delivered to the lung. The MMADs for Formulations XII-XV ranged from 3.42 to 4.76, indicating deposition in the central and conducting airways. [Table 34]

[0285] The weight loss of Formulations VII-VIII was measured by TGA and is detailed in Table 32. [Table 35]

[0286] The itraconazole content of Formulations VII and VIII was determined by HPLC-UV and is detailed in Table 33. [Table 36]

[0287] The crystallinity of Formulation XII was assessed by XRD. The diffraction pattern of itraconazole was observed in the formulation, suggesting that the milling or spray drying process did not affect the solid state of itraconazole. Additional peaks observed in the pattern correspond to additional excipients in the formulation (Figure 6).

[0288] The crystallinity of Formulation XIII was assessed by XRD. The diffraction pattern of itraconazole was observed in the formulation, suggesting that the milling or spray drying process does not affect the solid state of itraconazole. Additional peaks observed in the pattern correspond to additional excipients in the formulation (Figure 7).

[0289] The crystallinity of Formulation XIV was assessed by XRD. The diffraction pattern of itraconazole was observed in the formulation, suggesting that the milling or spray drying process does not affect the solid state of itraconazole. Additional peaks observed in the pattern correspond to additional excipients in the formulation (Figure 8).

[0290] The crystallinity of Formulation XV was assessed by XRD. The diffraction pattern of itraconazole was observed in the formulation, suggesting that the milling or spray drying process does not affect the solid state of itraconazole. Additional peaks observed in the pattern correspond to additional excipients in the formulation (Figure 9).

[0291] Example 8. Polysorbate 80-stabilized crystalline itraconazole dry powder formulation containing sodium sulfate / leucine and reduced levels of polysorbate 80 A. Powder preparation Formulation XVI, microcrystalline itraconazole, was prepared using a jet mill, Qualification Micronizer (Sturtevant, Hanover, MA, USA). The feed pressure was set at 90 psig, and the milling pressure was set at 40 psig. Itraconazole (SMS Pharma, Lot ITZ-0715005) was continuously fed into the mill until approximately 60 g of itraconazole was milled. The final median particle size (Dv(50)) of the milled API was approximately 1510 nm.

[0292] Formulation XVI, microcrystalline itraconazole, was then blended into a suspension in deionized water consisting of 10 wt% itraconazole and 0.25 wt% polysorbate 80. The batch size was 440 g. The polysorbate 80 was dissolved in 89.75% DI water with a magnetic stir bar, and then the micronized itraconazole was slowly added and mixed until the suspension was observed to be visually dispersed and uniform.

[0293] A feedstock solution was prepared and used to manufacture a dry powder composed of nanocrystalline itraconazole, polysorbate 80, and other additional excipients. A drug loading of 50 wt% itraconazole on a dry basis was targeted. The feedstock solution used to spray-dry the particles was made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and the solution was stirred until visually clear. The itraconazole-containing suspension was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock volume was 3000 g, which supported an approximately 1-hour manufacturing campaign. Table 34 lists the components of the feedstock used to prepare the dry powder. [Table 37]

[0294] A dry powder of Formulation XVI was produced from this raw material by spray drying in a Niro Mobile Minor spray dryer (GEA Process Engineering Inc., Columbia, MD) and collected in a bag filter. The system was operated in open-loop (single pass) mode using nitrogen as the drying and atomizing gas. Atomization of the feed liquid utilized a Schlick 940-0 atomizer equipped with a 1.0 mm liquid insert. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0295] The dry powders were produced according to the following spray-drying conditions: For Formulation XVI, the solids concentration of the liquid feed was 1.2%, the inlet temperature of the process gas was 181°C to 185°C, the outlet temperature of the process gas was 65°C, the flow rate of the drying gas was 80 kg / hr, the flow rate of the atomizing gas was 250 g / min, the back pressure of the atomizing gas at the atomizer inlet was 30.4 psig to 31.4 psig, and the flow rate of the liquid feed was 50 mL / min. The resulting dry powders are recorded in Table 35. [Table 38]

[0296] B. Powder Characterization The bulk particle size characteristics of the formulations are found in Table 36. The span at 1 bar of 1.93 for Formulation XVI indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersibility ratio of 1.03 for Formulation XVI indicates that the particle size is relatively insensitive to dispersion energy, a desirable feature that allows for similar dispersion over a range of dispersion energies. [Table 39]

[0297] The weight loss of Formulation XVI was determined by TGA and was found to be 0.37%.

[0298] The crystallinity of Formulation XVI was assessed by XRD. The diffraction pattern of itraconazole was observed in the formulation, suggesting that the milling or spray drying process did not affect the solid state of itraconazole. Additional peaks observed in the pattern correspond to additional excipients in the formulation. (Figure 10)

[0299] Example 9. Spray-dried dry powder formulation of itraconazole, sodium sulfate, and leucine A. Powder preparation A feedstock solution using a water-tetrahydrofuran (THF) cosolvent system was prepared and used to manufacture a dry powder composed of itraconazole, sodium sulfate, and leucine. A drug loading of 50 wt% itraconazole on a dry basis was targeted. The feedstock solution used to spray-dry the particles was prepared as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and the solution was stirred until visually clear. The required amount of THF was weighed into an appropriately sized glass container. The itraconazole was added to the THF and the solution was stirred until visually clear. The itraconazole THF solution was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock volume was 5 L, which supported an approximately 8.5-hour manufacturing campaign. Table 37 lists the components of the feedstock used to prepare the dry powder. [Table 40]

[0300] Dry powder of Formulation XIX was produced from this feedstock by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), with the powder collected in a cyclone. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. A Buchi nozzle with a 1.5 mm cap and a 0.7 mm liquid tip was used for atomization of the feed. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0301] The dry powders were produced according to the following spray-drying conditions: For Formulation XIX, the solids concentration of the liquid feed was 12.0 g / L, the inlet temperature of the process gas was 92°C-103°C, the outlet temperature of the process gas was 40°C, the drying gas flow rate was 17.0 kg / hr, the atomizing gas flow rate was 2830 g / min, and the liquid feed flow rate was 10.0 mL / min. The resulting dry powders are recorded in Table 38 below. [Table 41]

[0302] B. Powder Characterization The bulk particle size characteristics of the formulations are found in Table 39. The span at 1 bar of 2.32 for Formulation XIX indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersibility ratio of 1.12 for Formulation XIX indicates that the particle size is relatively insensitive to dispersion energy, a desirable feature that allows for similar dispersion over a range of dispersion energies. [Table 42]

[0303] The simulated patient flow rates of measured and / or calculated geometric particle size and capsule emitted powder mass (CEPM) at 60 liters per minute (LPM) and 30 LPM were measured for this formulation and are recorded in Table 40. The small changes in CEPM and geometric size from 60 LPM to 20 LPM indicate that the dry powder is relatively unaffected by patient inspiratory flow, resulting in relatively similar therapeutic doses delivered to patients breathing at various flow rates. [Table 43]

[0304] The aerodynamic particle size, fraction of fine particles, and fine particle dose measured and / or calculated by the Next Generation Impactor (NGI) are recorded in Table 41. The fine particle dose of Formulation XIX indicates that a high percentage of the nominal dose loaded into the capsule reaches the impactor stage (41.1%) and is therefore predicted to be delivered to the lung. The MMAD of Formulation XIX was 3.80 micrometers, indicating deposition in the central and conducting airways. [Table 44]

[0305] The weight loss of Formulation XIX was determined by TGA and was found to be 0.37%.

[0306] The itraconazole content of Formulation XIX was determined by HPLC-UV to be 99.0%.

[0307] The crystallinity of Formulation XIX was assessed by XRD (Figure 11). No itraconazole peaks were observed, indicating that no appreciable levels of itraconazole were present in the formulation. As shown, all peaks observed in the formulation correspond to excipients. Therefore, the solid-state itraconazole in Formulation XIX can be characterized as amorphous.

[0308] Example 10. In vitro dissolution test of dry powder formulations containing itraconazole A. In vitro dissolution test An in vitro model was used to develop a predictive study to understand itraconazole dissolution. Drug dissolution is a prerequisite for cellular uptake and / or absorption via the lungs. Therefore, itraconazole dissolution kinetics plays an important role in determining the extent of its absorption from the respiratory tract. For itraconazole-containing dry particles delivered as an aerosol to the respiratory tract, the fate of itraconazole in these particles is determined by the physicochemical properties of the particles. For itraconazole in aerosolized dry particles to exert its local effect in the lungs, the dry particles must first dissolve itraconazole, which must then be present in the lung fluid and lung tissue. However, once itraconazole dissolution into the lung fluid occurs, it can become more available for permeation and systemic absorption. The dissolution rate of itraconazole was predicted to be proportional to its solubility, concentration in the surrounding liquid film, and solid-liquid interfacial area. Solubility is determined by the drug's compounding, formulation, and physical form. The total liquid volume in the lungs is 10-30 mL, and the capsular liquid volume is approximately 5 μL / cm 2 , this volume may impair the solubilization and subsequent absorption of poorly soluble molecules, such as itraconazole.

[0309] The following in vitro dissolution model was used to understand the dissolution characteristics of dry powder aerosols containing itraconazole: Aerosol particles of the invention were collected using a Next Generation Impactor (NGI) (Copley Scientific, UK) at well-defined aerosol particle size distribution (APSD) cutoffs, and then their dissolution behavior was simulated using model lung fluid.

[0310] The aerosol dose collection system UniDose™ (Nanopharm, Newport, United Kingdom) in combination with a modified Next Generation Impactor (NGI) was used to uniformly deposit the impactor stage mass (ISM) (defined as the dose collected below stage 2 of the Next Generation Impactor) onto a filter suitable for subsequent dissolution testing in a USP V, paddle-over-disk (POD) apparatus.

[0311] B. Materials and Methods for In Vitro Dissolution Testing The materials used in this study are shown in Table 42. The powder formulation, capsules, and packaging materials were equilibrated at 22.5±2.5°C and 30±5% RH. The formulations were encapsulated in No. 3 HPMC capsules under the same conditions. The fill weight of the powder preparation was 10 mg. The formulations were aerosolized from the capsules using a capsule-based unit-dose DPI device (RS01, Plastiape, Osnago, Italy). One capsule of each formulation was aerosolized at 60 L / min (4 L inhalation volume) using a Plastiape RS01 dry powder inhaler (DPI). The aerosol dose was collected using a UniDose system. One milliliter of suspension formulation was aerosolized into a cNGI at 15 L / min using a Micro Mist™ Nebulizer (Hudson RCI, Temecula, CA, USA). Using the UniDose collection system, the entire impactor stage mass (i.e., less than stage 2 of the NGI) was uniformly deposited onto a glass microfiber filter membrane, visible as the location of the circle (impacting particle or droplet). The filter was placed in a disk cassette and dissolution tests were performed in a USP Apparatus II POD (paddle-over-disk, USP V) at 37°C using 500 ml of PBS (pH 7.4) and 2.0% SDS. Sink conditions were maintained within the container for all tests. Samples were taken at predetermined time points and tested for drug content on an Agilent (Santa Clara, CA, USA) 1260 Infinity Series HPLC. Data are presented as raw cumulative mass and cumulative mass percentage at 240 minutes (min). [Table 45]

[0312] C. Impactor Stage Mass (ISM) UniDose POD Dissolution Test Results of Formulations of the Present Invention A plot of the raw cumulative mass of ISM for the formulations is shown in Figure 12, and a plot of the cumulative mass dissolution (%) is shown in Figure 13. The UniDose ISM and dissolution half-life for each dry powder are summarized in Table 42. The particle size of the itraconazole crystals in suspension and the specific surface area (SSA) of the itraconazole crystals, estimated using particle size distribution measurements, are also shown in Table 43.

[0313] Based on the cumulative mass data, the collected ISM of the formulation ranged from 2.1 to 2.6 mg of itraconazole. These data suggest that the aerosolization efficiency of the formulation was approximately 50% based on the nominal dose. The itraconazole loading in each particle was 50%, and the nominal dose was 5 mg of itraconazole (10 mg of powder).

[0314] Formulation XIX had the fastest dissolution rate, with over 80% of the drug dissolved within the first time point. Due to the rapid dissolution kinetics of Formulation XIX, the dissolution half-life could not be calculated. The dissolution half-lives of the other dry powders indicated a ranking of their dissolution kinetics as follows: XI>XII>XIII>XIV>XV>Pure ITZ

[0315] The data presented in Figures 12 and 13 also evaluated the relationship between particle size of Formulations XI, XII, XIII, XIV, and XV and their respective dissolution half-lives, as shown in Figure 14A. These data suggest a good correlation between particle size and dissolution half-life of itraconazole crystals. Figure 14B shows the relationship between the specific surface area of ​​itraconazole crystals and dissolution half-life. These data suggest that increasing the surface area of ​​particles in the formulation shortens the dissolution half-life. These data demonstrate that the particle size, and thus the surface area, of the drug substance affects the dissolution behavior of the formulation.

[0316] Based on the pharmacokinetic data presented in Example 14, Formulation XIX had the highest systemic exposure. This correlated with its dissolution data, suggesting that this formulation has fast dissolution kinetics. The dissolution half-lives of other dry powders and the C of Formulation XIX were significantly higher than those of the other dry powders. max The relationship between is shown in Figure 15, or C max C expressed as a response rate max The relationship between dissolution half-life and C is shown in Figure 16. max There was an inverse correlation between the C and the systemic response of Formulation XIX, suggesting that the faster the dissolution rate, the higher the systemic exposure. maxThe correlation of the ratios strengthened with dissolution half-life. These data suggest that the systemic exposure response of an itraconazole formulation is modulated by its dissolution behavior and, therefore, the physicochemical properties of the formulation. [Table 46]

[0317] Raw cumulative mass % dissolution plots of ISM determined by UniDose POD for nanosuspension and microsuspension formulations are shown in FIG.

[0318] The dissolution rate of the nanosuspension was faster than that of the microsuspension: the dissolution half-life of the nanosuspension was 5.3 minutes and that of the microsuspension was 35.5 minutes.

[0319] Example 11. In vitro dissolution and permeability studies of dry powder formulations containing crystalline itraconazole A. In vitro elution and permeability testing A biorelevant dissolution test system was developed based on mimicking the air-liquid interface at the airway epithelial interface using a cell-based in vitro method. Materials were uniformly deposited onto the cell culture medium using a modified next-generation impactor (cNGI) incorporating a cell culture plate onto a collection stage. Drug elution and permeation through the epithelial cell monolayer were measured.

[0320] B. Materials and Methods for In Vitro Elution and Permeability Studies Epithelial cell monolayers grown at the air-liquid interface in permeable inserts, Snapwell™ (Corning Costar, Massachusetts, USA), were incorporated into cNGIs. Calu-3 cell lines (ATCC, LGC Standards, Teddington, UK) (passages 32-50) were grown in minimum essential medium (MEM) supplemented with non-essential amino acids, 10% (v / v) fetal bovine serum, 1% (v / v) penicillin-streptomycin, and 1% (v / v) antifungal agent Fungizone, and maintained at 37°C in a humidified atmosphere of 95% / 5% air / CO2, respectively. Cells were plated onto the Snapwell inserts, 1 cm thick. -2 5x10 5 Cells were seeded at a density of 1000 μm and cultured under air-interface conditions for 12 days, starting on day 2. Transepithelial electrical resistance (TEER) was measured using an EVOM2 chopstick electrode connected to an EVOM2 Epithelial Voltohmmeter (World Precision Instruments, Hitchin, United Kingdom). TEER was 450 Ω cm. 2 A monolayer that was greater than 100% was considered confluent.

[0321] The Snapwell containing Calu-3 ALI cells was transferred to the cup of a modified NGI and placed in stage 4 of the NGI (Copley Scientific, Nottingham, UK). One capsule of the dry powder of the present invention was aerosolized into the cNGI at 60 L / min for 4 seconds. One milliliter of the suspension was aerosolized into the cNGI at 15 L / min using a Micro Mist Nebulizer (Hudson RCI, Temecula, CA, USA).

[0322] The materials used in this study are shown in Table 41. The dry powder, capsules, and packaging materials were equilibrated at 22.5±2.5°C and 30±5% RH. The formulations were encapsulated in No. 3 HPMC capsules under the same conditions. The fill weight of the powder preparation was 10 mg. The formulations were aerosolized from the capsules using a capsule-based unit-dose DPI device (RS01™, Plastiape, Osnago, Italy). One capsule of each formulation was aerosolized at 60 L / min (4 L inhalation volume) using the Plastiape RS01™ dry powder inhaler (DPI).

[0323] After the dose was deposited onto the Snapwell from stage 4, the Snapwell was transferred to a 6-well plate containing 2 mL of PBS (pH 7.4) and 2.0% SDS maintained at 37°C. Basolateral samples were taken at various time points, and drug content was measured using an Agilent (Santa Clara, CA, USA) 1260 Infinity Series HPLC. The total amount delivered to the cells was determined from the total amount of drug dissolved over the time course and from lysed cells after the experiment.

[0324] C. Results of dissolution and permeability testing of powdered itraconazole formulations incorporated into cNGI The cumulative mass percent total recovery plot of dry powder containing itraconazole delivered to cells above stage 4 is shown in Figure 18. These data suggest differences in dissolution and permeation kinetics of the various formulations. Pure itraconazole as received had slower dissolution and permeation kinetics than the other formulations, while Formulation XIX had the fastest dissolution and permeation kinetics.

[0325] To understand the cNGI data for various formulations, we used the data to calculate the diffusion rate of the drug substance by taking into account the difference in loading. This calculation was performed using the following equation:

number

[0326] where J is the flux (slope of the cNGI dissolution / permeability profile), A is the barrier area, and C is the loading volume. These data are summarized in Table 44 and show that the diffusion rates of the formulations followed the following rank order: XIX>XI>XII>XIII>XIV>XV>Pure ITZ [Table 47]

[0327] Based on the pharmacokinetic data shown in Example 14, Formulation XIX had the highest systemic exposure. This correlates with the diffusion rate of this formulation, suggesting that this formulation has fast dissolution and permeation kinetics. The diffusion rates and C of the other dry powders were significantly higher than those of the other dry powders. max The relationship between the diffusion rate of formulation XIX and that of other dry powders is shown in Figure 19. max C expressed as a response rate max The relationship between the diffusion rate and C is shown in Figure 20. max The effect of C on the systemic response of Formulation XIX was suggested to be related, with the faster the diffusion rate, the higher the systemic exposure. max The correlation of the ratios became stronger with the diffusion rate.

[0328] The cNGI dissolution plots of the raw cumulative mass (%) of ISM for the nanosuspension and microsuspension formulations are shown in Figure 21. The cNGI data suggest that the diffusion rate of the nanosuspension formulation was faster than that of the microsuspension formulation.

[0329] Example 12. PK study of a single inhaled dose in rats A. Materials and Methods To assess the systemic exposure of male rats to itraconazole and its metabolite, hydroxy-itraconazole, at a nominal dose level of 5 mg / kg, blood and lung tissue samples were collected from rats after a single inhalation dose of each of five different itraconazole formulations over a 60-minute exposure period. Plasma concentrations of itraconazole and hydroxy-itraconazole were measured by a validated LC-MS / MS method in samples collected at the end of the exposure period and up to 96 hours after the end of exposure.

[0330] B. Results (plasma) The maximum mean plasma concentration of itraconazole (C max ) and the area under the mean plasma concentration versus time curve (AUC) estimated up to the last quantifiable time of the sample. last ) are summarized in Table 45. [Table 48]

[0331] C corrected for dose differences max and AUC last Based on the values ​​of , the highest mean plasma concentration (C max ) and the area under the mean plasma concentration versus time curve (AUC last ) for each group. max and average AUC last The ratios are shown in Table 46. [Table 49]

[0332] The rate of systemic exposure (C) of rats to itraconazole max ) and degree (AUC last ) was highest after exposure to Formulation XIX. max and AUC last was similar after exposure to Formulations XII and XI and slightly lower after exposure to Formulation XIV. max and AUC lastwas lowest after exposure to Formulation XV. A similar pattern was observed for the rate and extent of systemic exposure to hydroxy-itraconazole, but C max and AUC last The values ​​of were lower after exposure to formulation XII than after exposure to formulation XI and slightly higher after exposure to formulation XIV.

[0333] C. Results (lung tissue) The maximum mean lung tissue concentration of itraconazole (C max ) and the area under the mean lung tissue concentration versus time curve (AUC ) extrapolated to the last quantifiable time of the sample. last ) are summarized in Table 47. [Table 50]

[0334] C corrected for dose differences max and AUC last Based on the values ​​of , the highest mean lung tissue concentration (C max ) and the area under the mean lung tissue concentration versus time curve (AUC last ) for each group. max and average AUC last The ratios are shown in Table 48. [Table 51]

[0335] The rate of local lung exposure to itraconazole in rats (C max ) and degree (AUC last ) was lowest after exposure to Formulation XIX. max and AUC last was generally similar after exposure to Formulations XII, XI, and XIV, but AUC after exposure to Formulation XII last was somewhat lower than after exposure to the other two formulations. After exposure to formulation XV, C max was only slightly higher after exposure to formulation XIX and was lower than the values ​​for the other formulations, and AUC lastwas higher than after exposure to Formulation XIX and was roughly comparable to that after exposure to the other formulations. max and AUC last The values ​​were highest after exposure to Formulation XIX and lower after exposure to Formulations XII, XI, XIV and XV, but were roughly similar for all four of these formulations.

[0336] Lung AUC last The ratios of values ​​to the corresponding values ​​in plasma are shown in Table 49. [Table 52]

[0337] For itraconazole, the lung tissue to plasma ratio was lowest after exposure to Formulation XIX, similar after exposure to Formulations XII and XI, and somewhat higher after exposure to Formulation XIV. The highest ratio was observed after exposure to Formulation XV. For hydroxy-itraconazole, the lung tissue to plasma ratio was similar after exposure to each formulation and significantly lower than the ratio observed for itraconazole.

[0338] conclusion Systemic exposure to itraconazole in rats was highest after administration of Formulation XIX. Systemic exposure was similar after inhalation administration of Formulations XII and XI and slightly lower after administration of Formulation XIV. Systemic exposure was lowest after administration of Formulation XV. A similar pattern was observed for systemic exposure to hydroxy-itraconazole, with systemic exposure after administration of Formulation XII being lower than after administration of Formulation XI and slightly higher than after administration of Formulation XIV.

[0339] Local exposure to itraconazole in the lungs of rats was lowest after exposure to Formulation XIX. Local exposure was generally similar after administration of Formulations XII, XI, and XIV. After administration of Formulation XV, the maximum concentration was only slightly higher than after administration of Formulation XIX, but was lower than the values ​​for the other formulations, and the AUC lastValues ​​were higher after exposure to Formulation XIX and roughly similar to those after exposure to the other formulations. Topical exposure to hydroxy-itraconazole was highest after administration of Formulation XIX and lower after administration of Formulations XII, XI, XIV, and XV, but roughly similar for all four of these formulations.

[0340] Example 13. Amorphous Itraconazole Dry Powder Formulation Prepared for Use in a 28-Day Toxicity Study A. Powder preparation A feedstock solution using a water-tetrahydrofuran (THF) cosolvent system was prepared and used to manufacture a dry powder composed of itraconazole, sodium sulfate, and leucine. A drug loading of 50 wt% itraconazole on a dry basis was targeted. The feedstock solutions used to spray-dry the particles were prepared as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and the solution was stirred until visually clear. The required amount of THF was weighed into an appropriately sized glass container. The itraconazole was added to the THF and the solution was stirred until visually clear. The itraconazole THF solution was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. Each feedstock volume was 9.5625 L. Fourteen of these feedstocks were prepared for a total of 133.875 L, which supported an approximately 30-hour manufacturing campaign. Table 50 lists the ingredients of each raw material used to prepare the dry powder. [Table 53]

[0341] A dry powder of Formulation XX was produced from this raw material by spray drying in a Niro Mobile Minor spray dryer (GEA Process Engineering Inc., Columbia, MD) and collected in a bag filter. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. Atomization of the feed liquid utilized a Niro atomizer equipped with a 1.0 mm liquid insert. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0342] The dry powders were produced according to the following spray-drying conditions: For Formulation XX, the solids concentration of the liquid feedstock was 12 g / L, the inlet temperature of the process gas was 120°C to 140°C, the outlet temperature of the process gas was 40°C, the flow rate of the drying gas was 80 kg / hr, the flow rate of the atomizing gas was 352.2 g / min, the back pressure of the atomizing gas at the nebulizer inlet was 45 psig to 57 psig, and the flow rate of the liquid feedstock was 75 mL / min. The resulting dry powders are recorded in Table 51. The itraconazole in the formulation was amorphous. [Table 54]

[0343] B. Powder Characterization The bulk particle size characteristics of the formulations are found in Table 52. The span at 1 bar of 1.83 for Formulation XX indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersibility ratio of 1.06 for Formulation XX indicates that the particle size is relatively insensitive to dispersion energy, a desirable feature that allows for similar dispersion over a range of dispersion energies. [Table 55]

[0344] The weight loss of Formulation XX was determined by TGA and was found to be 0.34%.

[0345] The itraconazole content of Formulation XX was determined by HPLC-UV to be 100.9% of the nominal amount.

[0346] Example 14. Dry Powder Formulation of Crystalline Itraconazole Prepared for Use in a 28-Day Toxicity Study A. Powder preparation Formulation XXI, nanocrystalline itraconazole, was prepared as a suspension containing 25 wt% itraconazole (SMS Pharma lot ITZ-0715005) and 2.5 wt% polysorbate 80. The polysorbate 80 was dissolved in 72.5% deionized water using a magnetic stir bar, and then the itraconazole was added and suspended by stirring with a magnetic stir bar. Once all the itraconazole was suspended, the formulation was processed in a Netzsch MiniCer using 0.2 mm milling media (TOSOH, Tokyo, Japan) at 90% chamber fill. The itraconazole suspension was manufactured using the following conditions: mill speed 3000 RPM, inlet pump speed 100 RPM, recirculating chiller 10 °C, inlet air pressure 4.5 bar, and run time 30–40 min. Eight suspensions were generated in this manner and combined to form the final suspension lot. The final median particle size (Dv(50)) of the milled suspension was 130 nm.

[0347] Nanocrystalline itraconazole (Formulation XXII) was prepared as a suspension containing 10 wt% itraconazole, 0.7 wt% oleic acid, and 1.5% ammonium hydroxide in deionized water. The oleic acid was dissolved in 87.8% deionized water using a magnetic stirrer, followed by the addition of ammonium hydroxide and dissolution using a magnetic stirrer. Finally, itraconazole was added and mixed with a magnetic stirrer to form a suspension. Once all the itraconazole was suspended, the formulation was processed in a Netzsch MiniCer using 0.5 mm milling media (TOSOH, Tokyo, Japan) at 90% chamber fill. The itraconazole suspension was manufactured using the following conditions: mill speed 3000 RPM, inlet pump speed 100 RPM, recirculating chiller 10°C, inlet air pressure 4.5 bar, and run time 200–240 min. Eight suspensions were generated in this manner and combined to form the final suspension lot. The final median particle size (Dv(50)) of the milled suspension was 115 nm.

[0348] Formulation XXIII microcrystalline itraconazole was prepared using a jet mill, Qualification Micronizer (Sturtevant, Hanover, MA, USA). The feed pressure was set at 85 psig, and the milling pressure was set at 45 psig. Itraconazole was continuously fed into the mill until 480.0 g of itraconazole was milled. The final median particle size (Dv(50)) of the milled API was 1640 nm. Formulation XXIII micronized itraconazole was then blended into a suspension consisting of 10 wt% itraconazole and 0.25 wt% polysorbate 80 in deionized water. The batch size was 4800 g. The polysorbate 80 was dissolved in 88.75 g deionized water using a magnetic stir bar, and then the itraconazole was slowly added and mixed until the suspension was visually dispersed and uniform.

[0349] A feedstock suspension was prepared and used to manufacture a dry powder composed of crystalline itraconazole and other additional excipients. A drug load of 50 wt% itraconazole on a dry basis was targeted. The feedstock suspension used to spray-dry the particles was made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and stirred until the solution was visually clear. The itraconazole-containing suspension was then added to the excipient solution and stirred until visually homogenous. The feedstock was then spray-dried. The feedstock was stirred while spray-drying. Each individual feedstock mass for Formulation XXI was 7.5 kg. Six batches of these feedstocks were spray-dried, and these feedstocks supported a 15-hour manufacturing campaign. Each individual feedstock mass for Formulation XXII was 6.0 kg. Three batches of these feedstocks were spray-dried, and these feedstocks supported a 6-hour manufacturing campaign. Each individual feedstock mass for Formulation XXIII was 8.0 kg. Four of these feedstocks were spray dried and supported an approximately 11 hour manufacturing campaign. Tables 53 and 54 list the components of the feedstocks used to prepare the dry powders. [Table 56] [Table 57]

[0350] Dry powders of Formulations XXI-XXIII were produced from these raw materials by spray drying in a Niro Mobile Minor spray dryer (GEA Process Engineering Inc., Columbia, MD) and collected in a bag filter. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. Atomization of the feed liquid utilized a Niro two-fluid nozzle atomizer with a 1.0 mm liquid insert. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0351] The dried powders were produced according to the following spray-drying conditions: For Formulations XXI and XXII, the solids concentration of the liquid feed was 3%, the process gas inlet temperature was 170-190°C, the process gas outlet temperature was 65°C, the drying gas flow rate was 80.0 kg / hr, the atomizing gas flow rate was 250.0 g / min, and the liquid feed flow rate was 50.0 g / min. For Formulation XXIII, the solids concentration of the liquid feed was 1.2%, the process gas inlet temperature was 170-190°C, the process gas outlet temperature was 65°C, the drying gas flow rate was 80.0 kg / hr, the atomizing gas flow rate was 250.0 g / min, and the liquid feed flow rate was 50.0 g / min. The resulting dry powders are recorded in Table 55. [Table 58]

[0352] B. Powder Characterization The bulk particle size characteristics of the three formulations can be seen in Table 56. The span at 1 bar of less than 2.05 for Formulations XXI-XXIII indicates a relatively narrow size distribution. The 1 bar / 4 bar dispersion ratio of 1.25 for Formulations XXI-XXIII indicates that they are relatively independent of dispersion energy, a desirable property that allows for similar particle dispersion across a range of dispersion energies. [Table 59]

[0353] The weight loss of Formulations XXII-XXIII was measured by TGA and is detailed in Table 57. [Table 60]

[0354] The itraconazole content of Formulations XXI and XXIII was determined by HPLC-UV and is detailed in Table 58. [Table 61]

[0355] Example 17. 28-day inhalation toxicity studies A and B in rats A. Materials and Methods Two separate 28-day studies were conducted to evaluate both plasma and lung pharmacokinetics and potential local tissue toxicity. In the first study, 28-day Study A, five groups of rats received either air or placebo control or one of three doses of Formulation XX daily for 28 days. In the second study, 28-day Study B, seven groups of rats received one of three crystalline nanoparticulate itraconazole formulations daily or, in the case of Group 1, every third day for 28 days. The groups and achieved doses are detailed in Tables 59 and 60. [Table 62] [Table 63]

[0356] In both studies, blood and lung tissue samples were collected from male and female rats after the first and final inhalation doses of each formulation to assess pulmonary and systemic exposure and accumulation of itraconazole. Additionally, lung tissue samples, including larynx, trachea, carina, and lung, were collected from all rats 24 hours after the final dose to assess treatment-related microscopic pathological changes. Plasma and lung concentrations of itraconazole in the samples were measured using a validated LC-MS / MS method.

[0357] B. Results (plasma) The maximum mean plasma concentrations (C) of itraconazole were observed in male and female rats on days 1 and 28. max ), and the area under the mean plasma concentration versus time curve (AUC) estimated up to the last quantifiable time of the sample. 0-last ) are summarized in Table 61 for amorphous itraconazole in 28-day Study A and in Table 62 for crystalline itraconazole in 28-day Study B. [Table 64] [Table 65]

[0358] Despite the differences in the absolute amounts achieved between the various formulations, the peak (C max ) and total (AUC 0-last It is clear that the systemic exposure is higher than either of the crystalline formulations after both a single dose (Day 1) and multiple doses (Day 28). Table 63 below shows the mean C values ​​for itraconazole from both 28-day studies for each formulation, using a target dose of 15 mg / kg / day for each study. max and AUC 0-last Dose-normalized values ​​are summarized. Normalization was performed by dividing the measured exposure by the actual achieved dose for each day of each study. [Table 66]

[0359] Systemic exposure of itraconazole was measured in rats on day 1 at C max and AUC 0-last Dose-normalized values ​​for C were highest after exposure to Formulation XX. By Day 28, Formulation XX still generally had a higher C max and AUC 0-last The dose-normalized values ​​of AUC were higher for crystalline formulation XXI, especially in females, but the difference was not significant. In males, the AUC 0-last were slightly higher. These data indicate that for a given achieved lung delivery, the systemic exposure resulting from inhalation of the crystalline formulation is generally less than that of Formulation XX. However, considering that systemic exposure is determined by both the lung tissue dissolution rate and lung tissue permeability of the material in the lung, the crystalline formulation also demonstrates sufficient dissolution and tissue permeation over time to reduce or eliminate the difference, providing confidence that the crystalline formulation is not simply an insoluble lung deposit.

[0360] C. Results (lung tissue) The mean lung tissue trough concentrations (23 hours after the end of the previous dose) on days 1 to 28 for each group, expressed as a percentage of the corresponding mean plasma concentration values ​​at the same time points, are shown in Table 64. [Table 67]

[0361] For itraconazole, lung tissue to plasma ratios were lowest after exposure to Formulation XX and were consistently significantly higher for all crystalline formulations on both Day 1 and Day 28. These data indicate that the crystalline formulations provide substantially higher lung exposures at lower systemic exposures at the doses tested, enhancing exposure at the site of action while minimizing the potential for undesirable systemic exposure effects.

[0362] D. Results (lung pathology) In the 28-day Study A, microscopic findings related to Formulation XX were present in respiratory tissues at doses above 5 mg / kg / day. Minimal to mild granulomatous inflammation was present at all doses, with macrophages and multinucleated giant cells frequently containing intracytoplasmic spicules. The highest dose included a 28-day recovery period, but these only partially resolved. The documented pathology was considered adverse at all doses because it was scattered and did not completely resolve during the recovery period. The spicules noted in the pathology were likely itraconazole, and we theorize that they form when amorphous material supersaturates the lung capsule fluid and interstitial space after multiple doses, leading to crystallization of the API. Shorter duration exposure studies with the same formulation did not demonstrate these findings.

[0363] In the 28-day Study B, Formulations XXI and XXIII were associated with minimal adverse accumulation of foamy macrophages in the lungs only at 40 mg / kg / day (Formulation XXI was the only formulation administered at that level). There were no apparent differences in the incidence or severity of findings among rats administered Formulations XXI-XXIII at similar dose levels. Overall, the no-observed-adverse-effect level (NOAEL) was approximately 15 mg / kg / day for all three crystalline formulations tested.

[0364] Pathological findings associated with the amorphous composition in the rat airways were distinct in character from those induced by the crystalline formulation, and findings in the latter group were more related to a clearance response to accumulated material in the airway lumen as opposed to granulomatous inflammation within the airway mucosa. Additionally, findings associated with the amorphous formulation affected more areas of the airways and were deleterious at lower doses.

[0365] conclusion Systemic exposure, i.e., plasma levels, to itraconazole in rats was highest after administration of Formulation XX. Systemic exposure was generally lower after inhalation administration of Formulations XXI–XXIII, but by 28 days of administration, the difference diminished after a single dose. However, lung exposure was significantly and consistently higher for Formulations XXI–XXIII compared to Formulation XX. When comparing lung and systemic exposure, the ratio for Formulation XX was higher in lung than in systemic exposure. However, lung to plasma ratios were substantially higher for each of the crystalline formulations, XXI–XXIII. These data indicate that the crystalline formulations delivered substantially higher local concentrations of itraconazole and resulted in systemic exposures less than those of Formulation XX.

[0366] The amorphous nature of itraconazole in Formulation XX results in increased solubility and rapid penetration into the systemic circulation through the lungs, as evidenced by significantly higher systemic exposure on Day 1. Administration of Formulation XX also resulted in local toxicity in the form of spicules in the mucosa and adverse granulomatous inflammation at all doses tested, even at doses as low as 5 mg / kg / day. The use of crystalline nanoparticles in Formulations XXI-XXIII resulted in substantially greater lung retention and higher local exposures than the amorphous formulations at similar or lower systemic exposures. This altered exposure profile has the advantage of increasing pulmonary efficacy while potentially further minimizing, rather than worsening, the undesirable effects of itraconazole systemic exposure for Formulation XX. Additionally, Formulations XXI-XXIII were associated with significantly lower potential for adverse microscopic pathology findings despite substantially higher local exposures.

[0367] Overview of in vitro and in vivo examples Investigation of the impact of itraconazole's physical form in dry powders involved iterative progression through in vitro dissolution and permeability studies, as well as in vivo single- and multiple-dose pharmacokinetic and toxicity studies. In vitro dissolution studies demonstrated that the physical form and particle size of itraconazole within the formulation played an important role in determining dissolution rate and the expected rate at which delivered material clears the lungs and penetrates into the systemic circulation. These data demonstrate the ability to control important aspects of both pulmonary and systemic exposure, allowing for modulation of both efficacy and, potentially, adverse outcomes. These in vitro findings were validated in an in vivo single-dose inhalation PK study, confirming that powders containing crystalline itraconazole nanoparticles, when delivered by inhalation, exhibited prolonged lung retention and, after a single dose, a higher lung-to-plasma ratio and lower peak and total systemic exposure compared to formulations containing amorphous itraconazole. The outlined example of a 28-day inhalation toxicity study further demonstrates that distinct exposure kinetics are maintained for amorphous itraconazole in dry powder and crystalline itraconazole with respect to pulmonary and systemic exposure over multiple days after dosing. Additionally, examination of the amorphous and crystalline material for microscopic pathology after multiple days of dosing reveals differences in both the nature and severity of these findings, with the crystalline material exhibiting fewer adverse findings and only higher pulmonary exposure.

[0368] Example 18. Human Simulation: Oral Inhalation and Oral Solution Administration Certain assumptions were made for this human simulation. The lung systemic absorption rate estimated using a rat model was used as input for the human simulation. The lung solubility value from the rat model was used as the starting point for the human simulation. Particle size distribution (MMAD and GSD) data using Alberta Idealized Throat was used in GastroPlus™ with the ICRP66 model to estimate human deposition fractions. A practical dose was used that resulted in approximately 56% deposition in the lungs and 12.6% deposition in the throat, with the remaining percentage of the drug estimated to be retained in the device.

[0369] Single-dose pharmacokinetic parameters for Formulation XII were simulated over 14 days of repeated exposure. Dose-proportional increases in both total lung and plasma concentrations were predicted from 5 mg to 20 mg. Similar half-lives were predicted in the lung and plasma. [Table 68]

[0370] Proportional increases in plasma and lung amounts are predicted after multiple doses. Seven days after dosing, the model predicted accumulation in the lung and even greater accumulation in plasma. Based on human predictions, some accumulation of undissolved drug in the interstitial regions of the alveoli was expected with subsequent doses. Plasma concentrations after oral solution administration were higher than those at either the 5 mg or 20 mg oral inhalation dose levels. However, total lung concentrations were higher after oral inhalation administration. Thus, the total lung to plasma ratio was significantly higher after oral inhalation administration compared with oral solution administration. [Table 69]

[0371] Table 67 shows modeled human clinical data following a single dose of Formulation XII inhaled (oral inhalation) at 5 mg or 20 mg doses, or 200 mg of oral SPORANOX® (oral solution). For lung to plasma ratios, lung and plasma AUC data are compared for each dose over a 7-day period; the ratio is substantially higher for inhaled administration than for oral administration. While oral administration may achieve lung levels capable of producing therapeutic lung levels, it would require a larger total dose delivered and a higher systemic exposure. Without wishing to be bound by theory, it is believed that the same lung exposure using 0.2 mg inhalation would be achieved with 200 mg orally. [Table 70]

[0372] Table 68 shows the exposure over a 24-hour period at "steady state" on Day 21. Daily dosing by inhalation was compared to possible dosing every other day (EOD) by inhalation. The EOD dosing option appeared to be half the daily dose, so it may be possible to improve exposure kinetics based on the regimen. Even with EOD dosing, lung exposure is significantly higher than that seen after oral dosing of 200 mg daily. [Table 71]

[0373] Figures 22A and 22B show the kinetics of three itraconazole-containing formulations at a 5 mg dose. On the left (Figure 22A), a graph shows plasma exposure during routine clinical use comparing a twice-daily dosing regimen of SPORANOX® with once-daily dosing of either Formulation XIX or Formulation XII. Quite clearly, inhaled dosing resulted in significantly lower systemic exposure, with Formulation XII ultimately achieving trough exposure levels similar to those of Formulation XIX, but with less daily variability and a C max reached that level at a fairly low level.

[0374] On the right (Figure 22B), lung exposure for the same dose and regimen is shown; the dotted line approximates the Aspergillus MIC (approximately 500 ng / g or ng / mL). With oral administration, lung levels reach levels above this MIC, but only for a short period between twice-daily doses, and for the majority of the exposure period, exposure is below "effective" levels for treating fungal infections. Formulation XIX and SPORANOX® at 5 mg had very similar lung exposure profiles. However, even Formulation XII, the lowest dose of 5 mg, had an exposure profile that consistently exceeded the MIC and SPORANOX® over the entire 24-hour period on Day 1 and throughout the 7-day administration period. Itraconazole's antifungal efficacy is based on AUC / MIC, meaning that exposure above the MIC, both in terms of total exposure and time, is a key factor in determining efficacy. Clearly, Formulation XII achieves a theoretical exposure that contributes significantly more to antifungal efficacy than either Formulation XIX or oral SPORANOX®, both of which significantly reduce systemic exposure. Without wishing to be bound by theory, these results demonstrate that the dry powders disclosed herein, including an angiogenesis inhibitor (e.g., itraconazole), can be used to achieve and / or maintain therapeutically effective concentrations of the angiogenesis inhibitor (e.g., itraconazole) in lung tissue, which may be useful in treating cancer, such as lung cancer, including NSCLC. In particular, studies of treating NSCLC in humans with high-dose oral itraconazole have shown a direct and significant correlation with reduced tumor volume and tumor perfusion, reduced angiogenesis-stimulating cytokines IL1b and GM-CSF, and reduced tumor microvascular density (Gerber et al. Clin. Cancer Res. (2020) 26:6017-6027).

[0375] Figures 23A and 23B show the kinetics of three itraconazole formulations at a 20 mg dose. The results and interpretation are similar to those for the 5 mg inhalation dose described above for Figures 22A and 22B, except that for inhalation, the dose is higher and the corresponding lung and plasma exposures are higher. Formulation XIX, with the higher dose, achieves a lung exposure above the MIC and higher than SPORANOX® over the 24-hour period. Conversely, the plasma exposure remains significantly below that of SPORANOX®. The 20 mg exposure of Formulation XII results in a higher lung exposure than the 5 mg dose, which consistently remains above the MIC and above that of SPORANOX® throughout the entire time course.

[0376] Example 19. Phase 1 / 1b: Safety-Tolerability Study Safety, tolerability, and PK studies in healthy volunteers and asthma patients demonstrate lung and plasma PK advantages over oral SPORANOX®. In Part 1 of the study, single ascending doses (5 mg, 10 mg, 25 mg, and 35 mg) of Formulation XII were administered to normal healthy volunteers (n=6 / cohort). In Part 2 of the study, multiple ascending doses (10 mg, 20 mg) of Formulation XII were administered to healthy volunteers (n=6 / cohort), with an optional third cohort receiving a dose up to 35 mg. The safety and tolerability of Formulation XII were evaluated during administration for up to 14 days, at doses expected to produce lung exposures greater than fivefold higher than oral SPORANOX® and itraconazole plasma levels greater than fivefold lower than those observed with oral SPORANOX®.

[0377] Part 3 of the study evaluated the safety and tolerability of Formulation XII or oral SPORANOX® administered as a single dose to asthma patients (n=16) in a crossover design. Patients who received 200 mg of oral SPORANOX® in the first period received a 20 mg dose of Formulation XII in the second period, and patients who received 20 mg of Formulation XII in the first period received 200 mg of oral SPORANOX® in the second period. Sputum and plasma itraconazole levels were measured to assess lung and plasma exposure. This study confirmed that lung exposure with Formulation XII resulted in lung concentrations higher than the minimum inhibitory concentration (MIC) for A. fumigatus and higher than those achieved with oral SPORANOX®. The plasma exposure of itraconazole following administration of Formulation XII was more than five-fold lower than that observed with oral SPORANOX®. Without wishing to be bound by theory, these results are believed to indicate that the dry powders disclosed herein, including crystalline itraconazole, can be used to achieve and maintain therapeutically effective concentrations of itraconazole in lung tissue for treating lung cancer (e.g., NSCLC), while minimizing plasma concentrations and reducing the risk of toxicity and side effects.

[0378] Example 20: Comparison of airway findings from two rat and three dog studies with inhalation exposure to inhaled itraconazole formulations XIX and XII Studies were conducted in rats and dogs in two laboratories using inhalation of itraconazole-containing dry powders formulated using spray drying. All studies contained the same active pharmaceutical ingredient, but the formulation excipients in some cases, and in particular the physiochemical properties of the itraconazole in the particles, varied. The studies and results are summarized below.

[0379] Testing in rats Formulation XIX was administered in rats for a 28-day inhalation study, followed by a 28-day recovery period.

[0380] Rats were exposed to air, placebo, or itraconazole formulated as Formulation XIX (itraconazole was at a 50% formulation concentration) at target doses of 5 mg / kg / day, 20 mg / kg / day, or 44 mg / kg / day for 28 days. Microscopic findings related to Formulation XIX were present in the lungs, bronchi, larynx, and carina at doses of 5 mg / kg / day and above, and in the trachea at doses of 20 mg / kg / day. Minimal to mild granulomatous inflammation was present in the lungs and bronchi at doses of itraconazole 5 mg / kg / day and above. The granulomatous inflammation was characterized by aggregates of macrophages and multinucleated cells within the bronchiolar mucosa, often forming papillary protrusions in the lumen. The macrophages and multinucleated giant cells frequently contained intracytoplasmic spicules. Alveolar macrophage aggregates were also present in the lungs at an incidence above background in rats dosed at 20 mg / kg / day and above. These macrophages were vacuolated, resulting in a foamy appearance of cytoplasm.

[0381] In the larynx and tracheal carina, minimal to mild granulomatous inflammation was present at doses of 5 mg / kg / day or higher of itraconazole. As in the lung, this inflammation was characterized by intracytoplasmic spicules within the mucosa, along with aggregates of macrophages and multinucleated giant cells. Similar minimal granulomatous inflammation was present in the tracheal mucosa of rats treated with 20 mg / kg / day or higher.

[0382] At the end of the 28-day recovery period, granulomatous inflammation of the bronchioles was still present in rats dosed with 44 mg / kg / day, suggesting that the bronchiolar findings at this dose did not resolve during the recovery period. Granulomatous inflammation was not observed in the larynx, carina, or trachea at the end of the recovery period, suggesting that it had completely resolved during the recovery period in these tissues.

[0383] In summary, the primary finding associated with Formulation XIX was granulomatous inflammation characterized by mucosal macrophages with cytoplasmic spicules and multinucleated giant cells. This finding was observed in rats receiving itraconazole at doses of 5 mg / kg / day and above, but was considered adverse at all doses. It was present throughout the conducting airways, from the larynx to the small bronchioles, and did not resolve in the bronchioles during the recovery period in rats treated with 44 mg / kg / day (other dose groups were not examined at the end of the recovery period). Aggregates of alveolar macrophages with foamy cytoplasm were also present in the lungs at an incidence above background at the time of terminal sacrifice in rats treated with 20 mg / kg / day and above.

[0384] 28-day inhalation study in rats with Formulation XII or Formulation XV Rats were exposed to itraconazole formulated as Formulation XII at target doses of 5 mg / kg / day, 15 mg / kg / day, or 40 mg / kg / day, or to Formulation XV at doses of 5 mg / kg / day or 15 mg / kg / day, for 28 days. In both cases, itraconazole was 50% of the total formulation concentration. In addition, one group of rats received 15 mg / kg of itraconazole as Formulation XII every 3 days. Minimal to mild accumulation of foamy macrophages associated with Formulation XII and Formulation XV was present in the lungs at 15 mg / kg / day, with the incidence and severity being higher in rats receiving Formulation XII at 40 mg / kg / day. Minimal accumulation of foamy macrophages was observed in the lungs with Formulation XII or Formulation XV at 5 mg / kg / day administered every 3 days, or with Formulation XII at 15 mg / kg / day; however, due to the lack of air- or placebo-control rats, it was not possible to determine whether there were test-item-related effects at these doses. Mild subacute inflammation, considered test-item related and adverse, was present in rats administered Formulation XII at 40 mg / kg / day. Minimal subacute inflammation was observed in rats administered Formulation XII or Formulation XV at 15 mg / kg / day; however, it was unclear whether the inflammation was test-item related. There were no clear differences in the incidence or severity of macrophage accumulation or subacute inflammation between male rats administered Formulation XII and Formulation XV at the corresponding dose levels. There was evidence that the severity and / or incidence of these findings was higher in female rats administered Formulation XV at 15 mg / kg / day compared with Formulation XII.

[0385] Dog testing A 7-day inhalation study (with a 14-day recovery period) of Formulation XIX in dogs. Dogs were exposed to itraconazole formulated as Formulation XIX at target doses of 5 mg / kg / day, 10 mg / kg / day, or 20 mg / kg / day for 7 days. The itraconazole formulation concentration was 50% of the total. A 14-day recovery group was included for dogs exposed to 5 mg / kg / day. Minimal to mild acute inflammation associated with Formulation XIX was considered adverse and was present in both dogs (one male, one female) treated with 20 mg / kg / day, and minimal acute inflammation was present in the female dog treated with 10 mg / kg / day. The acute inflammation was characterized by the presence of neutrophils, macrophages, and some multinucleated giant cells that appeared to contain spicules in their cytoplasm (observed upon post-study slide review). Thus, the acute inflammation exhibited characteristics of granulomatous inflammation. No test-item-related findings were observed in dogs treated with 5 mg / kg / day at terminal or recovery sacrifice.

[0386] A 28-day inhalation study of Formulation XIX in dogs with a 28-day recovery period. Dogs were exposed to air, placebo, or formulated itraconazole at target doses of 5 mg / kg / day, 10 mg / kg / day, or 20 mg / kg / day for 28 days. The itraconazole formulation concentration was 50% of the total. Minimal to mild bronchiolar / peribronchiolar granulomatous inflammation associated with Formulation XIX was present in both males and females at doses of 5 mg / kg / day and above. The incidence and severity of this finding increased with treatment in males at doses of 5 mg / kg / day and above and in females at 20 mg / kg / day. The granulomatous inflammation was located in and around the terminal and respiratory bronchioles and was characterized by aggregates of macrophages and multinucleated giant cells (rich in eosinophilic cytoplasm). Mild granulomatous inflammation was observed at doses of 10 mg / kg / day and above and was considered adverse. The granulomatous inflammation completely resolved during the recovery period.

[0387] A 14-day inhalation study of Formulations XII and XV in dogs. Dogs were exposed for 14 days to placebo or itraconazole formulated as Formulation XII at target doses of 2 mg / kg / day, 6 mg / kg / day, or 20 mg / kg / day, or as Formulation XV at target doses of 6 mg / kg / day or 20 mg / kg / day. Additionally, one group of dogs received itraconazole as Formulation XII at a target dose of 6 mg / kg every three days. Itraconazole formulation concentrations were 50% of the total in all cases. Test-item-related airway findings were present in dogs receiving Formulation XII or Formulation XV at 20 mg / kg / day. Test-item-related mild intraalveolar mixed cell inflammation was present in all dogs receiving Formulation XII at 20 mg / kg / day. Test-item-related mild lymphocytic inflammation of the carina and tracheal mucosa was present in two of three dogs receiving Formulation XII at 20 mg / kg / day. Additionally, minimal mixed cell inflammation within the alveoli was present in one of three dogs receiving 20 mg / kg / day Formulation XV. Thus, the location of findings varied somewhat between Formulation XII and Formulation XV. While this variability complicated comparisons between Formulation XII and Formulation XV, the dose level (20 mg / kg / day) at which findings clearly related to the test article were observed was the same for both test articles. Mild mixed cell inflammation was present in one of three dogs receiving 6 mg / kg of Formulation XII every three days and one of three dogs receiving 6 mg / kg / day Formulation XV. The low incidence of this finding in each of these groups and the absence of findings in other airway segments at these doses made the relationship to inhalation of Formulation XII or Formulation XV unclear.

[0388] Comparison of Formulation XIX with Formulations XII and XV in rat studies Formulation XIX-related findings in the rat airways were characterized differently from those induced by Formulation XII and Formulation XV. In addition, Formulation XIX-related findings affected more tissues in the airways and appeared to be deleterious at lower doses. Recovery was not assessed in rat studies with Formulation XII or Formulation XV. However, based on experience with other inhalants, granulomatous inflammation within the bronchiolar mucosa, while present after exposure to Formulation XIX, appears to resolve more slowly than the increase in alveolar macrophages or subacute inflammation induced by Formulation XII or Formulation XV.

[0389] Granulomas, or granulomatous inflammation, composed of macrophages and multinucleated giant cells are a common response to substances that are not readily solubilized in cytoplasmic lysosomes, including inhaled foreign matter. The presence of these cells at multiple levels in the mucosa of the respiratory tract after inhalation of Formulation XIX suggests that the test article exerted its effects and either a) penetrated the epithelium and was phagocytosed by macrophages, or b) lysed, penetrated the epithelium, recrystallized in the interstitium, and was phagocytosed by macrophages in the interstitium, or b) lysed, penetrated the epithelium, recrystallized in the interstitium, and was phagocytosed by macrophages in the interstitium. It is not surprising that poorly soluble forms of substances do not completely resolve during the recovery period.

[0390] Exposure to Formulation XII at 40 mg / kg / day resulted in mild subacute inflammation, which was considered test article-related and adverse. This subacute inflammation was observed in the alveolar parenchyma and was morphologically distinct from the granulomatous mucosal inflammation observed with Formulation XIX exposure. Minimal subacute inflammation was observed in rats receiving Formulation XII or Formulation XV at 15 mg / kg / day, but it was unclear whether this was test article-related. When comparing Formulation XII and Formulation XV, there were no clear differences in the incidence or severity of macrophage accumulation or subacute inflammation between male rats receiving these test articles at the corresponding dose levels. Female rats receiving Formulation XV at 15 mg / kg / day suggested a higher severity and / or incidence of these findings compared with Formulation XII. Recovery was not investigated in studies with Formulation XII or Formulation XV. However, minimal to mild subacute inflammation and minimal to mild macrophage accumulation were considered reversible findings and were generally expected to resolve over the 28-day recovery period.

[0391] It is difficult to compare alveolar macrophage accumulation across these rat studies because studies of Formulation XII or Formulation XV did not include air or placebo controls. Different groups of rats in different study laboratories may have different background incidences of minimal alveolar macrophage accumulation. However, data from these studies suggest that minimal to mild alveolar macrophage accumulation may have occurred at higher incidences and / or lower doses in rats treated with Formulation XII or Formulation XV, possibly indicating superior alveolar distribution of the macrophage-phagocytosed form of Formulation XII or Formulation XV.

[0392] Comparison of Formulation XIX with Formulations XII and XV in Dog Studies In the dog respiratory tract, findings associated with Formulation XIX had characteristics that differed from those induced by Formulations XII and XV.

[0393] Acute inflammation associated with Formulation XIX, which was considered adverse, was present in the 7-day dog ​​study at 10 mg / kg / day. There were no findings associated with Formulation XIX at 5 mg / kg / day in the 7-day study. Granulomatous inflammation associated with Formulation XIX was present at doses above 5 mg / kg / day in the 28-day dog ​​study, reaching mild severity and considered adverse at doses above 10 mg / kg / day. In retrospect, the acute inflammation observed in the 7-day study could be described as acute granulomatous inflammation. Thus, the findings were similar across the two dog studies with Formulation XIX, although the differences reflected the length of the studies. In both studies, the findings were considered adverse at 10 mg / kg / day. The location of the inflammation was primarily bronchiolar / peribronchiolar, as opposed to intraalveolar. This location indicates the orientation of the conducting airways, somewhat similar to the location of findings associated with Formulation XIX in rats, but the mucosal location was absent and less discreet than in rats.Granulomatous inflammation was similar, but not morphologically identical, between rats and dogs.

[0394] Formulation XII or Formulation XV induced test-item-related findings at 20 mg / kg / day in a 14-day study in dogs. Mild intra-alveolar mixed cell inflammation related to the test item was present in all dogs receiving Formulation XII at 20 mg / kg / day. No adverse events were noted in the 14-day study records; however, mild mixed cell inflammation would have been considered adverse. Findings at 6 mg / kg / day were clearly not test-item-related due to their low incidence. Dogs were not exposed to Formulation XII or Formulation XV at 10 mg / kg / day, so direct comparisons with Formulation XIX (which was adverse at 10 mg / kg / day) are not possible. However, when comparing adverse events after inhalation exposure, it may be more appropriate to compare lung tissue levels, as opposed to doses, which were substantially higher in animals treated with Formulation XII and Formulation XV. Granulomatous inflammation associated with Formulation XIX completely resolved during the 28-day recovery period. There was no recovery period in the 14-day dog ​​studies with Formulation XII and Formulation XV. The mixed cell inflammation associated with Formulation XII or Formulation XV in dogs was somewhat morphologically similar to the subacute inflammation associated with Formulation XII or Formulation XV in rats, which affected the alveoli and was not granulomatous.

[0395] Example 21: A Phase 1, Open-Label Study to Evaluate the Safety, Tolerability, and Pharmacokinetics of Single and Multiple Doses of Itraconazole Administered as a Dry Powder for Inhalation in Healthy Subjects Clinical pharmacokinetics of itraconazole oral solution based on historical data Itraconazole is metabolized in the liver by the isoenzyme cytochrome P450 3A4 to the major metabolite hydroxy-itraconazole. Itraconazole is highly bound by plasma proteins (99.8% for oral solution and 99.6% for capsules).

[0396] The pharmacokinetics of oral itraconazole have been studied in humans in both single-dose and multiple-dose studies. The pharmacokinetics differed between the two forms (solution and capsules), with higher exposures observed in the oral solution. It is recommended that the oral solution and capsules not be used interchangeably.

[0397] The absolute bioavailability of its oral solution is 55% in healthy volunteers and increases by 30% when taken under fasting conditions. Under fasting conditions, the steady-state AUC 0-24 h is 131 ± 30% of the exposure under fed conditions, and it is recommended that the oral solution be administered in a fasted state. At steady state, under fasting conditions, the mean C max was 1,963±601ng / mL, and the mean t max was 2.5±0.8 hours, and the mean AUC 0-24h The mean elimination rate of itraconazole was 29,271 ± 10,285 ng·h / mL. The half-life of itraconazole at steady state was 39.7 ± 13 hours.

[0398] Pharmacokinetic data from Part 1 (SAD) in healthy volunteers Summary systemic pharmacokinetic data following a single inhaled dose of Formulation XII are summarized in Table 69, and the concentration versus time profile is shown in Figure 24. The pharmacokinetic data are significant due to their impact on the safety profile of Formulation XII compared with oral administration. The data confirm that inhaled administration of Formulation XII results in low systemic exposure. Doses ranged from 5 mg to 35 mg of itraconazole. Itraconazole was rapidly absorbed into the systemic circulation, with detectable plasma exposure in all subjects at the earliest sampling time of 15 minutes. Exposure was generally maintained over the first 18 to 24 hours, indicating prolonged absorption. Beyond 24 hours post-dose, plasma concentrations declined steadily and generally monoexponentially, with decay rates similar across all cohorts (K el Geometric mean range: 0.021-0.032 1 / h). Exposure (C max and AUC) increased monotonically and generally less than dose-proportional. [Table 72]

[0399] Pharmacokinetic data from part 2 (MAD) in healthy volunteers Summary systemic pharmacokinetic data after a single inhaled dose and 14 days of daily inhalation of Formulation XII are summarized in Table 70, and concentration versus time profiles are shown in Figure 25. Doses were either 10 mg, 20 mg, or 35 mg of itraconazole. As in Part 1, itraconazole was rapidly absorbed and entered the systemic circulation, with detectable plasma exposure in all subjects at the earliest sampling time of 15 minutes. Exposure was generally sustained over the first 18-24 hours, with T max The median estimates ranged from 7 to 18 hours across the cohort.

[0400] Median plasma concentrations increased with each repeated dose, and concentrations approached steady state by day 14. Compared with steady-state plasma levels of itraconazole recorded after oral solution administration, exposure after inhalation was significantly higher than AUC 0-24h Between days 1 and 14, the accumulation of itraconazole was 100-400 times lower than that of C max and AUC 0-24h Both concentrations were approximately three-fold and similar at each dose. As in Part 1, at the end of dosing, plasma concentrations declined steadily monoexponentially, suggesting no significant lung accumulation that would prolong systemic exposure. [Table 73]

[0401] Pharmacokinetic data from Part 3 (single dose) in adults with mild to moderate stable asthma Summary systemic pharmacokinetic data after a single inhaled or oral dose in asthma patients are summarized in Table 71, and concentration versus time profiles are shown in Figures 26A and 26B. The dose was either 20 mg itraconazole inhaled as Formulation XII or 200 mg itraconazole administered as SPORANOX® Oral Solution. With both oral and inhaled administration, itraconazole is rapidly absorbed and enters the systemic circulation, resulting in T max The estimated median exposure was 4.0 hours for Formulation XII and 1.5 hours for SPORANOX®. After Formulation XII administration, itraconazole plasma exposure generally increased and / or was maintained over the first 24 hours, indicating prolonged absorption. In contrast, orally administered itraconazole was rapidly absorbed and eliminated, with exposure peaking shortly after administration but decreasing to a 17% Cmax at 12 hours post-dose. max The total systemic exposure (AUC 0-24h ) after formulation XII was approximately 85-fold lower than the exposure after oral administration, and the highest exposure (C max ) after formulation XII was approximately 250-fold lower than exposure after oral administration. [Table 74]

[0402] Induced sputum was collected 2, 6, and 24 hours after administration and used to measure itraconazole concentrations using a validated liquid chromatography-mass spectrometry / mass spectrometry (LC-MS / MS) method with an LLOQ (lower limit of quantitation) of 0.1 ng / mL. Sputum itraconazole levels were higher with Formulation XII compared with oral SPORANOX®, with a geometric mean C max was 5381ng / mL, whereas C maxThe mean lung exposure was 116.3 ng / mL (Figure 26A). The high lung exposure after Formulation XII was maintained over a 24-hour period, with sputum itraconazole concentrations declining 2-6 hours after a single 200 mg oral dose of itraconazole. These data confirm that inhaled administration of Formulation XII results in high and sustained lung exposure, higher than that achieved with oral administration, while maintaining low systemic exposure. The geometric mean C in lung and plasma max Based on the data, Formulation XII resulted in a lung:plasma ratio of approximately 2300:1, while oral administration resulted in a lung:plasma ratio of 1:5.

[0403] Example 22. Dry powder formulations containing crystalline microparticle form of itraconazole at various drug loads Nanocrystalline itraconazole formulations XXXXI–XXXXVI were prepared as suspensions containing 35.0 wt% itraconazole (SMS Pharma, Lot ITZ-0715005) and 2.92 wt% polysorbate 80, with the itraconazole contained at a 12:1 ratio (wt:wt) to polysorbate 80. The polysorbate 80 was dissolved in 62.1% deionized water using a magnetic stir bar, and then the itraconazole was added and suspended by stirring with a magnetic stir bar. Once all the itraconazole was suspended, the formulations were processed in a Netzsch MiniCer using 0.2 mm grinding media (TOSOH, Tokyo, Japan) at 90% chamber fill. The itraconazole suspensions were manufactured using the following conditions: The mill speed was 3000 RPM, the inlet pump flow rate was 220 mL / min, the recirculation chiller was 10° C., and the run time was 37 minutes. The final median particle size (Dv(50)) of the milled suspension was 141 nm.

[0404] A raw material suspension was prepared and used to manufacture dry powders containing itraconazole in crystalline microparticle form and additional excipients. Drug loadings of 50 wt%, 60 wt%, 70 wt%, and 80 wt% itraconazole on a dry basis were targeted. The raw material suspensions used to spray-dry the particles were made as follows: The required amount of water was weighed into an appropriately sized glass container. The excipients were added to the water and stirred until the solution was visually clear. The itraconazole nanosuspension was then added to the excipient solution and stirred until visually homogeneous. The raw material was then spray-dried. The raw material was stirred while spray-drying. Table 72 lists the ingredients of the raw material used to prepare the dry powders. [Table 75]

[0405] Dry powders of Formulations XXXXI-XXXXVI were produced from the corresponding raw materials in Table 72 by spray drying in a Buchi B-290 Mini Spray Dryer (Buchi Labortechnik AG, Flawil, Switzerland), with the powder collected in a cyclone. The system was operated in open-loop (single-pass) mode using nitrogen as the drying and atomizing gas. A Schlick 970-1 nozzle was used for atomization of the feed solution. The system aspirator was adjusted to maintain a system pressure of -2.0 inches of water.

[0406] The dry powder was produced according to the following spray drying conditions: the solids concentration of the liquid feed was 30 g / kg, the drying gas flow rate was 17.0 kg / hr, the atomizing gas flow rate was 19.6 g / min, and the liquid feed flow rate was 3.0 mL / min. The inlet temperature of the process gas was varied to keep the outlet temperature constant at 65°C. The resulting dry powder formulations are recorded in Table 73. [Table 76]

[0407] B. Powder Characterization The bulk particle size properties of the six formulations are summarized in Table 74. The 1 bar / 4 bar dispersibility ratios of less than 1.1 and the 0.5 bar / 4 bar dispersibility ratios of less than 1.25 for Formulations XXXXI-XXXXVI indicate that they are relatively independent of dispersion energy, a desirable property that allows for similar particle dispersion across a range of dispersion energies. [Table 77]

[0408] The weight loss of Formulations XXXXI-XXXXVI was measured by TGA and is detailed in Table 7. [Table 78]

[0409] The aerodynamic particle size, fine particle fraction, and fine particle load for formulations XXXXI, XXXXIV, and XXXXVI, measured and / or calculated by Next Generation Impactor (NGI), are recorded in Table 76. The fine particle load for all formulations indicates that a high percentage of the nominal dose loaded into the capsule reaches the impactor stage (greater than 45%) and is therefore predicted to be delivered to the lung. The MMAD for all formulations was less than 3.5 μm, indicating deposition in the central and conducting airways. [Table 79]

Claims

1. A dried powder for use in the treatment of cancer (e.g., lung cancer, e.g., NSCLC), comprising homogeneous respiratory dried particles containing an angiogenesis inhibitor (e.g., itraconazole) and optionally a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., sodium salt and leucine), for administration to the target airway.

2. Use of a dried powder in the manufacture of a pharmaceutical product for the treatment of cancer (e.g., lung cancer, e.g., NSCLC), wherein the dried powder comprises homogeneous respiratory dried particles comprising an angiogenesis inhibitor (e.g., itraconazole) and optionally a stabilizer (e.g., polysorbate 80) and / or one or more excipients (e.g., sodium salt and leucine), and the pharmaceutical product is for administration to the target airway.

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