Compositions of surface-modified therapeutically active particles produced by ultra-rapid freezing

Abstract

The present invention describes pharmaceutical compositions comprising less than 10% excipients and present as nanoaggregates. These pharmaceutical compositions have been shown to exhibit improved properties, such as improved atomization and aerodynamic performance. The present invention also provides methods for preparing the pharmaceutical compositions disclosed herein and uses thereof.

Description

[0001] This application claims priority to U.S. Provisional Application No. 62 / 702,674, filed on July 24, 2018, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present disclosure relates generally to the field of pharmaceuticals and pharmaceutical manufacturing. More specifically, the present disclosure relates to compositions and methods of preparing pharmaceutical compositions comprising a small amount of excipients and a therapeutic agent and formulated as nanoaggregates. Background Art

[0003] Until recently, aerosolized antifungal drugs delivered to the lungs were limited to amphotericin B (Le and Schiller, 2010; Borro et al., 2008). However, Hilberg et al. reported in 2008 that inhaled voriconazole was more effective than inhaled amphotericin B in the treatment of invasive pulmonary aspergillosis (IPA), confirming the initial report by Tolman et al. (2009a) of aerosolized voriconazole formulations for the successful treatment of IPA patients who had previously failed oral or parenteral voriconazole with or without inhaled amphotericin B.

[0004] Tolman et al. reported the delivery of inhaled voriconazole to the lungs by nebulization (Tolman et al., 2009a; Tolman et al., 2009b). However, the concentration of voriconazole in lung tissue dropped below the minimum detectable concentration after 6 hours (Tolman et al., 2009a). In addition, the efficacy of the nebulized formulation was also very low, only 5.9% (w / w) when sodium sulfobutyl ether-β-cyclodextrin (SBECD) was used as an excipient. The safety of SBECD delivery via the pulmonary route has not been confirmed, and this high content of inactive ingredients may cause serious side effects (Wong 1993). Sinha et al. and Arora et al. reported the use of polylactide-co-glycolide nanoparticles (Sinha et al., 2013) and polylactide microparticles (Arora et al., 2015) to prepare voriconazole formulations for dry powder inhalation (DPI), respectively, but the drug loading of these particles was low, 31% w / w and 20% w / w, respectively). Arora et al. reported another voriconazole powder formulation for DPI prepared using leucine as an excipient (Arora et al., 2016). However, all of these DPI powder formulations contain non-GRAS excipients that have not been approved by the FDA for inhaled drugs. Beinbom et al. also used particle engineering technology (i.e., thin film freezing, TFF) to develop amorphous and crystalline voriconazole formulations suitable for dry powder inhalation (Beinbom et al., 2012a; Beinbom et al., 2012b). However, the amorphous formulation contained 75% (w / w) excipients, so the efficacy was low, and based on in vivo pharmacokinetic data in a mouse model, the drug absorption efficiency was low and the clearance rate was fast. In the lungs and plasma, the AUC of the crystalline formulation was 0-24h Both were significantly higher than the amorphous preparation, which were 452.6 μg·h / g and 232.1 μg·h / g in the lungs, and 38.4 μg·h / g and 18.6 μg·h / g in plasma, respectively. However, the aerosol performance of the crystalline preparation was poor (FPF 37.8%).

[0005] Recently, researchers have proposed based on models that nanoaggregates containing drug nanoparticles have more favorable distribution in the lungs and higher epithelial coverage than discrete micron-sized particles and nanoparticles (Longest and Hindle, 2017). Aggregates are solid materials in particulate form, consisting of a combination of particles held together by strong intermolecular or intramolecular cohesion (Chiou and Riegelman, 1971). When three different forms of particulate drugs (including conventional microparticles, nanoaggregates, and true nanoaerosols of budesonide and fluticasone propionate) were tested in a computational model, the total absorption efficiency of fluticasone propionate nanoaggregates was 57 times higher than that of conventional microparticles. Although true nanoaerosols have superior absorption efficiency, there is no actual device available for delivering true nanoaerosols to the small airways, so nanoaggregates provide the best approach for targeting drugs to the small airways. According to Longest et al., slowly dissolving nanoaggregates are believed to have improved drug absorption and distribution (Longest and Hindle, 2017).

[0006] TFF is a particle engineering technique that uses ultrafast freezing rates of up to 10,000 K / s (Engstrom et al., 2008). Due to the high degree of supercooling, TFF has been successfully used to produce nanostructured aggregates (Sinswat et al., 2008). Spray drying is another common technique for producing micro- or nano-sized particles for DPI. However, the formation of particles during the drying process of spray drying generally takes longer than the freezing process of TFF (Wisniewski 2015), resulting in longer particle growth time and larger particle size. Therefore, typical spray drying methods do not have the advantages of enhanced absorption and microdosing of nanoaggregates as described by Longest and Hindle (2017). Therefore, there is still a need to develop other pharmaceutical compositions as nanoaggregates that exhibit improved properties, such as enhanced atomization effects. Summary of the invention

[0007] The present disclosure provides a pharmaceutical composition comprising a therapeutic agent and an excipient as a nanoaggregate, a method for making the same, and a method for using the same. In some embodiments, the present disclosure provides a pharmaceutical composition comprising:

[0008] (A) a therapeutic agent; and

[0009] (B) an excipient, wherein the excipient comprises less than about 10% by weight of the pharmaceutical composition;

[0010] Wherein the pharmaceutical composition is formulated as nanoaggregates, the nanoaggregates comprise nanoparticles of the therapeutic agent, and the surfaces of the nanoparticles of the therapeutic agent comprise discrete domains of the excipient, and wherein the discrete domains of the excipient reduce the contact area between the nanoparticles of the therapeutic agent.

[0011] In some embodiments, the therapeutic agent exists in a crystalline state. In other embodiments, the therapeutic agent exists in an amorphous state. In some embodiments, the excipient comprises about 9% w / w to about 1% w / w of the pharmaceutical composition, such as about 6% w / w to about 2% w / w of the pharmaceutical composition. In some embodiments, the excipient comprises about 3% w / w of the pharmaceutical composition. In other embodiments, the excipient comprises about 5% w / w of the pharmaceutical composition.

[0012] In some embodiments, the discrete domains of the excipient include one or more discontinuous domains of the excipient on the surface. In other embodiments, the discrete domains of the excipient include adjacent and continuous excipient layers. In some embodiments, the excipient is soluble in water. In some embodiments, the excipient is solid at room temperature. In some embodiments, the excipient is a sugar alcohol, such as mannitol. In some embodiments, the excipient exists as a nano domain in the pharmaceutical composition. In some embodiments, the nano domain of the excipient has a size of about 50nm to about 500nm such as about 100nm to about 200nm.

[0013] In some embodiments, the pharmaceutical composition has a mass median aerodynamic diameter of about 1.5 μm to about 7.5 μm, such as about 2.5 μm to about 6.5 μm. In some embodiments, the pharmaceutical composition does not contain a wax excipient. In some embodiments, the pharmaceutical composition does not contain a hydrophobic excipient. In some embodiments, the therapeutic agent is selected from the group consisting of anticancer drugs, antifungal agents, psychotherapeutic agents such as analgesics, consciousness level altering agents such as anesthetics or hypnotics, nonsteroidal anti-inflammatory drugs (NSAIDS), anthelmintics, beta-agonists, anti-acne drugs, antianginal drugs, antiarrhythmic drugs, antiasthmatic drugs, antibacterial agents, anti-benign prostatic hypertrophy agents, anticoagulants, antidepressants, antidiabetic drugs, antiemetics, antiepileptic drugs, antigout drugs, antihypertensive drugs, anti-inflammatory drugs, antimalarials, antimigraine drugs, antimuscarinics, antineoplastic drugs, antiobesity drugs, antiosteoporotic drugs, In some embodiments, the therapeutic agent is an antifungal agent, such as an azole antifungal agent. In some embodiments, the therapeutic agent is an antifungal agent, such as an azole antifungal agent. In some embodiments, the azole antifungal agent is voriconazole. In some embodiments, the pharmaceutical composition further comprises one or more additional excipients. In some embodiments, the pharmaceutical composition further comprises one or more additional therapeutic agents.

[0014] In some embodiments, the pharmaceutical composition is formulated for use by the following routes: oral, intra-fat, intra-arterial, intra-articular, intracranial, intradermal, intralesional, intramuscular, intranasal, intraocular, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrarectal, intrathecal, intratracheal, intratumoral, intraumbilical, intravaginal, intravenous, intracapsular, intravitreal, intraliposome, topical, mucosal, parenteral, rectal, subconjunctival, subcutaneous, sublingual, topical, transdermal, vaginal, in the form of an emulsion, in the form of a lipid composition, via a catheter, via lavage, via continuous infusion, via infusion, via inhalation, via injection, via local delivery or via local perfusion. In some embodiments, the pharmaceutical composition is formulated for use via inhalation.

[0015] In certain embodiments, pharmaceutical composition is formulated for use with inhaler, and this inhaler is for example fixed dose combination inhaler, single dose dry powder inhaler, multi-dose dry powder inhaler, multi-unit dose dry powder inhaler, metered dose inhaler or pressurized metered dose inhaler.In certain embodiments, inhaler is capsule type inhaler (capsule-basedinhaler).In certain embodiments, inhaler is low resistance inhaler.In other embodiments, inhaler is high resistance inhaler.In certain embodiments, inhaler is used under the flow rate of about 10L / min to about 150L / min such as about 20L / min to about 100L / min.In certain embodiments, inhaler has the pressure difference of 0.5kPa to about 5kPa.In certain embodiments, the pressure difference is 1kPa, 2kPa or 4kPa.In certain embodiments, inhaler has the loading dose of about 0.1mg to about 50mg.In certain embodiments, inhaler has the loading dose of about 0.1mg to about 10mg.In other embodiments, inhaler has the loading dose of about 5mg to about 50mg such as about 5mg to about 25mg. In certain embodiments, inhaler is configured to deliver one or a series of doses from one or more unit doses that load successively. In certain embodiments, inhaler is configured to deliver a dose from a unit dose. In other embodiments, inhaler is configured to deliver a series of doses from a unit dose. In other embodiments, inhaler is configured to deliver a dose separately from a series of capsules that load successively. In other embodiments, inhaler is configured to deliver a series of doses from a series of capsules that load successively.

[0016] In another aspect, the present disclosure provides a method for treating or preventing a disease or illness in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition as described herein, the pharmaceutical composition comprising a therapeutic agent effective to treat the disease or illness. In some embodiments, the disease or illness is in the lungs. In some embodiments, the disease or illness is an infection, such as a fungal infection. In some embodiments, the therapeutic agent is an antifungal agent, such as an azole antifungal agent. In some embodiments, the therapeutic agent is voriconazole.

[0017] In another aspect, the present disclosure provides a method for preparing a pharmaceutical composition, comprising:

[0018] (A) mixing a therapeutic agent and an excipient with a solvent to form a precursor solution, wherein the excipient is present in an amount of less than 10% w / w;

[0019] (B) depositing the precursor solution onto a surface, the surface being at a temperature suitable to cause freezing of the solvent; and

[0020] (C) removing the solvent to obtain a pharmaceutical composition.

[0021] In some embodiments, the solvent is a mixture of two or more solvents. In some embodiments, the mixture of solvents comprises water. In some embodiments, the solvent is an organic solvent. In some embodiments, the organic solvent is acetonitrile. In other embodiments, the organic solvent is 1,4-dioxane. In some embodiments, the solvent is a mixture of water and an organic solvent, such as a mixture of water and acetonitrile. In some embodiments, a mixture of two or more solvents comprises about 10% v / v to about 90% v / v of the organic solvent. In some embodiments, the mixture comprises about 40% v / v to about 60% v / v of an organic solvent such as about 50% v / v of an organic solvent. In other embodiments, the mixture comprises about 20% v / v to about 40% v / v of an organic solvent such as about 30% v / v of an organic solvent. In some embodiments, the therapeutic agent and excipient account for less than 10% w / v of the precursor solution, such as about 0.5% w / v to about 5% w / v of the precursor solution. In some embodiments, the therapeutic agent and excipient account for about 1% w / v of the precursor solution. In other embodiments, the therapeutic agent and excipient comprise about 3% w / v of the precursor solution.

[0022] In some embodiments, the surface is rotating. In some embodiments, the temperature is about 0°C to about -200°C. In some embodiments, the temperature is about 0°C to about -120°C, such as about -50°C to about -90°C. In some embodiments, the temperature is about -60°C. In other embodiments, the temperature is about -125°C to about -175°C, such as about -150°C. In some embodiments, the solvent is removed under reduced pressure. In some embodiments, the solvent is removed via freeze drying. In some embodiments, freeze drying is carried out at a freeze drying temperature of about -20°C to about -100°C, such as about -40°C. In some embodiments, the reduced pressure is less than 250mTorr, such as about 100mTorr.

[0023] In some embodiments, the method further includes heating the pharmaceutical composition under reduced pressure. In some embodiments, the pharmaceutical composition is heated to a temperature of about 0°C to about 30°C, such as about room temperature or about 25°C. In some embodiments, the reduced pressure is less than 250mTorr, such as about 100mTorr. In some embodiments, the reduced pressure is the same as the reduced pressure during lyophilization.

[0024] In another aspect, the present disclosure provides a pharmaceutical composition prepared according to the method described herein.

[0025] Through the following detailed description, other purposes, features and advantages of the present disclosure will become apparent. However, it should be understood that although specific embodiments of the present invention are indicated, the detailed description and specific embodiments are given by way of illustration only, because through this detailed description, various changes and modifications within the spirit and scope of the present invention will become apparent to those skilled in the art. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The following drawings constitute a part of this specification and are included in this specification to further illustrate certain aspects of the present disclosure. The present disclosure may be better understood by referring to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0027] Figure 1 Shown are the XRPDs of (a) voriconazole powder; (b) TFF-VCZ; (c) TFF-VCZ-MAN 95:5; (d) TFF-VCZ-MAN 70:30; (e) TFF-VCZ-MAN 50:50; and (f) TFF-MAN.

[0028] Figure 2 Modulated DSCs of (a) TFF-MAN; (b) TFF-VCZ; (c) TFF-VCZ-MAN 95:5; (d) TFF-VCZ-MAN 50:50 are shown.

[0029] Figures 3A-3J show SEM images of TFF-VCZ-MAN: (Figure 3A) TFF-VCZ; (Figure 3B) TFF-VCZ-MAN 95:5; (Figure 3C) TFF-VCZ-MAN 70:30; (Figure 3D) TFF-VCZ-MAN 50:50; (Figure 3E) TFF-VCZ-MAN 25:75; (Figure 3F) TFF-MAN; (Figure 3G) atomized TFF-VCZ-MAN 95:5; (Figure 3H) atomized TFF-VCZ-MAN50:50; (Figure 3I) TFF-VCZ-MAN 25:75, after treatment in Franz cell for 5 min; (Figure 3J) TFF-VCZ-MAN 95:5, after treatment in Franz cell for 5 min.

[0030] Figures 4A-4F SEM images of (FIG. 4A) TFF-VCZ and (FIG. 4B) TFF-VCZ-MAN 95:5 are shown; (FIG. 4C) TFF-VCZ; (FIG. 4D) 3D morphology of TFF-VCZ-MAN 95:5; and (FIG. 4E) the contact area and distance between TFF-VCZ and (FIG. 4F) TFF-VCZ-MAN 95:5 particles are shown.

[0031] Figure 5 Shown is the AFM topography of atomized TFF-VCZ-MAN 95:5 obtained by a DP4 insufflator.

[0032] Figure 6 The SSA of TFF-VCZ-MAN powder formulations are shown (n=3; mean±SD).

[0033] Figures 7A-7C The SEM / EDX data of TFF-VCZ-MAN 50:50 are shown: (FIG. 7A) SEM image; (FIG. 7B) elemental analysis result of spot A; (FIG. 7C) elemental analysis result of spot B.

[0034] Figures 8A and 8B show the FT-IR (Figure 8A, 3500 cm -1 Up to 3100cm -1 region; Figure 8B, 1290 cm -1 Up to 1230cm -1 Area): (a) voriconazole powder; (b) TFF-VCZ; (c) TFF-VCZ-MAN 95:5; (d) TFF-VCZ-MAN 70:30; (e) TFF-VCZ-MAN 50:50; (f) TFF-MAN.

[0035] Figures 9A and 9B show the 1D CP-MAS spectra of (Figure 9A) TFF-VCZ and (Figure 9B) TFF-VCZ-MAN 90:10; 13 C spectrum (spectrum on the left) and 19 F spectrum (spectrum on the right).

[0036] 10A and 10B show the following 2D 1 H- 13 CHETCOR spectra: (FIG. 10A) TFF-VCZ; and (FIG. 10B) TFF-VCZ-MAN 90:10.

[0037] Fig.11 The FPF (percentage of quantitative dose) of TFF-VCZ-MAN dry powder formulations are shown (n=3; mean±SD).

[0038] Fig.12 Aerodynamic size distribution characteristics of TFF-VCZ-MAN 95:5 are shown by shear time from left to right: 0 min; 15 min; 30 min; 60 min (n=3; mean±SD).

[0039] Figures 13A and 13B show the aerodynamic properties of TFF-VCZ-MAN 95:5 by shear time: (a line) FPF, percent delivered dose; (b line) FPF, percent quantified dose; (c line) MMAD; and (d line) GSD (n=3; mean±SD).

[0040] Figures 14A and 14B show the aerodynamic properties of TFF-VCZ-MAN 95:5 according to storage time at 25°C / 60%RH: (line a) FPF, percentage of delivered dose; (line b) FPF, percentage of quantified dose; (line c) MMAD; and (line d) GSD (n=3; mean±SD).

[0041] Fig.15 The cumulative release rate (%) of voriconazole is shown: (line a) TFF-VCZ-PVPK25 25:75 (amorphous); (line b) TFF-VCZ-MAN 25:75; (line c) TFF-VCZ-MAN 50:50; (line d) TFF-VCZ-MAN 95:5 (n=3; mean±SD).

[0042] Fig.16 An image of the freezing process is shown.

[0043] Figures 17A-17D The following AFM topography images are shown: (a) Formulation #2 (size 5 μm x 5 μm) and (b) Formulation #4 (size 2 μm x 2 μm); and the corresponding following 3D topography images: (c) Formulation #2 and (d) Formulation #4.

[0044] Figures 18A-18F SEM images of voriconazole nanoaggregates are shown: (a) Formulation #1, (b) Formulation #2, (c) Formulation #3, (d) Formulation #4, (e) Formulation #5, and (f) Formulation #6.

[0045] Figures 19A-19F SEM images of aerosolized voriconazole nanoaggregates are shown: (a)-(b) Formulation #7 and (c)-(f) Formulation #6.

[0046] Fig. 20 The following XRPDs are shown: (a) voriconazole powder, (b) TFF-voriconazole, (c) Formulation #6 (small scale), (d) Formulation #6 (large scale), and (e) TFF-mannitol. DETAILED DESCRIPTION

[0047] In some aspects of the present disclosure, pharmaceutical compositions comprise nanoaggregates. These compositions can be prepared by methods such as thin film freezing and comprise a therapeutic agent and an excipient. In some embodiments, these compositions also exhibit modified atomization or provide other pharmaceutical properties.

[0048] Also provided herein are methods of making and using these compositions. The details of these compositions are described in detail below.

[0049] I. Pharmaceutical Compositions

[0050] In some aspects, the present disclosure provides pharmaceutical compositions comprising a therapeutic agent and an excipient, wherein the excipient comprises less than about 10% w / w of the composition. These pharmaceutical compositions may further comprise one or more additional therapeutic agents or one or more additional excipients. Such compositions may be prepared using methods such as thin film freezing. These methods comprise freezing a solution of the therapeutic agent and the excipient in a solvent and then removing the solvent under reduced pressure and / or cryogenic conditions. Methods for preparing pharmaceutical compositions using thin film freezing are described in: U.S. Patent Application No. 2010 / 0221343; Watts et al., 2013; Engstrom et al., 2008; Wang et al., 2014; Thakkar et al., 2017; O'Donnell et al., 2013; Lang et al., 2014a; Lang et al., 2014b; Carvalho et al., 2014; Beinbom et al., 2012a; Beinborn et al., 2012 b; Zhang et al., 2012; Overhoff et al., 2008; Overhoff et al., 2007a; Overhoff et al., 2007b; Watts et al., 2010; Yang et al., 2010; DiNunzio et al., 2008; Yang et al., 2008; Purvis et al., 2007; Liu et al., 2015; Sinswat et al., 2008; and U.S. Patent No. 8,968,786, which are incorporated herein by reference.

[0051] In pharmaceutical compositions as herein described, nano-aggregates can be used as nano-aggregates, and this nano-aggregates comprises the combination of nano-particles attracted or combined together by intermolecular or intramolecular cohesion.In pharmaceutical compositions as herein described, nano-aggregates can comprise one or more drug particles, and these drug particles are coated by the discrete discontinuous nano-domain of excipient.Do not wish to be bound by any theory, it is believed that the nano-domain of excipient can comprise about 25nm, 50nm, 75nm, 100nm, 125nm, 150nm, 175nm, 200nm, 225nm, 250nm, 275nm, 300nm, 325nm, 350nm, 375nm, 400nm, 425nm, 450nm, 475nm, 500nm, 525nm, 550nm, 575nm, 600nm, 625nm or 650nm or can be derived from the size of any scope therein. These nano-domains of excipients may include a size of about 25 nm to about 750 nm, about 50 nm to about 500 nm, or about 100 nm to about 200 nm. Without wishing to be bound by any theory, it is believed that these nano-domains may exist as discrete compositions that form dots on the surface of the nano-aggregates containing the therapeutic agent. The pharmaceutical composition may further comprise a mass median aerodynamic diameter of about 2.5 μm to about 7.5 μm, about 3.0 μm to about 6.0 μm, about 4.0 μm to about 6.0 μm, or about 2.5 μm, 2.75 μm, 3.0 μm, 3.25 μm, 3.5 μm, 3.75 μm, 4 μm, 4.25 μm, 4.5 μm, 4.75 μm, 5 μm, 5.25 μm, 5.5 μm, 5.75 μm, 6.0 μm, 6.25 μm, 6.5 μm, 6.75 μm, 7.0 μm, 7.25 μm, to about 7.5 μm, or any range derivative therein.

[0052] A. Therapeutic agents

[0053] "Therapeutic agent" used in the methods and compositions of the present invention refers to any substance, compound, drug, medicament or other major active ingredient that can provide a therapeutic or pharmacological effect when administered to a human or animal. When a therapeutic agent is present in the composition, the content of the therapeutic agent in the composition is between about 50% w / w to about 99% w / w, between about 70% w / w to about 99% w / w, between about 90% w / w to about 97% w / w, or between about 95% w / w to about 97% w / w of the total weight of the composition. In some embodiments, the amount of the therapeutic agent is about 50% w / w, 60% w / w, 70% w / w, 80% w / w, 90% w / w, 91% w / w, 92% w / w, 93% w / w, 94% w / w, 95% w / w, 96% w / w, 97% w / w to about 99% w / w or any range derived therefrom.

[0054] Suitable lipophilic therapeutic agents can be any poorly water-soluble bioactive agent or a salt, isomer, ester, ether or other derivative thereof, including but not limited to anticancer drugs, antifungal agents, psychiatric therapeutic agents such as analgesics, consciousness-altering agents such as anesthetics or hypnotics, nonsteroidal anti-inflammatory drugs (NSAIDS), anthelmintics, anti-acne drugs, anti-angina drugs, anti-arrhythmic drugs, anti-asthma drugs, antibacterial agents, anti-benign prostatic hypertrophy agents, anticoagulants, antidepressants, antidiabetics, antiemetics, antiepileptics, antigout drugs, antihypertensive drugs, anti-inflammatory drugs, antimalarials, antimigraine drugs, antimuscarinics, antineoplastics, antihypertensive drugs, Obesity drugs, osteoporosis drugs, Parkinson's disease drugs, antiproliferative agents, antiprotozoal drugs, antithyroid drugs, antitussive drugs, anti-incontinence drugs, antiviral agents, antianxiety agents, appetite suppressants, beta-agonists, beta-blockers, cardiac positive inotropes, chemotherapy drugs, cognitive enhancers, birth control pills, corticosteroids, Cox-2 inhibitors, diuretics, erectile dysfunction improvers, expectorants, gastrointestinal drugs, histamine receptor antagonists, immunosuppressants, keratolytics, lipid regulators, leukotriene inhibitors, macrolides, muscle relaxants, neuroleptics, nutritional supplements, opioid analgesics, protease inhibitors or sedatives.

[0055] Non-limiting examples of therapeutic agents can include 7-methoxypteridine, 7-methylpteridine, abacavir, abafungin, abarelix, acebutolol, naproxenol, acetaminophen, acetanilide, acetazolamide, acetohexamide, acitretin, acrivastine, adenine, adenosine, alatrofloxacin, albendazole, salbutamol, alclofenac, aldesleukin, alemtuzumab, alfuzosin, alitretinoin, alobarbital, allopurinol, all-trans retinoic acid (ATRA), aloprin, alprazolam, alprenolol, hexamethylmelamine, amifostine, amiloride, aminoglutethimide, aminopyrine, amiodarone hydrochloride, amitriptyline, amlodipine, amobarbital, Amodiaquine, amoxapine, amphetamine, amphetamine, amphotericin, amphotericin B, ampicillin, amprenavir, amsacrine, amyl nitrite, amobarbital, anastrozole, amrinone, anthracene, anthracyclines, aprobital, arsenic trioxide, asparaginase, aspirin, astemizole, atenolol, atorvastatin, atovaquone, atrazine, atropine, azathioprine, auranofin, azacitidine, azapropazone, azathioprine, azantamide, azithromycin, aztreonam, baclofen, barbiturates, live BCG, beclomethasone, bendrofluazide, benazepril, benidipine, benolate bevacizumab (Avastin), bexarotene, bezafibrate, bicalutamide, bifonazole, biperiden, bisacodyl, bisantrene, bleomycin, bleomycin, bortezomib, brinzolamide, bromazepam, bromocriptine mesylate, bromperidol, brotizolam, budesonide, bumetanide, bupropion, busulfan, butalbital, butamben, butenafine hydrochloride, butalbital, butalbital (Butesa), butoconazole, butoconazole nitrate, parahydroxybenzoic acid Butyl ester, caffeine, calcifediol, calcipotriol, calcitriol, captestosterone, canbendazole, camphor, camptothecin, camptothecin analogs, candesartan, capecitabine, capsaicin, captopril, carbamazepine, thyroxine, furadan, carboplatin, carbromide, thyroxine, carmustine, cefamandole, cefazolin, cefixime, ceftazidime, cefuroxime axetil, celecoxib, cephradine, cerivastatin, cetrizine, cetuximab, chlorambucil, chloramphenicol, chlordiazepoxide, clomethiazole, chloroquine, chlorothiazide, chlorpheniramine, chlorproguanil hydrochloride, chlorpromazine, chlorsulfuronamide, chlorprothixene, chlorpyrifos, chlortetracycline, chlorthalidone, chlorzoxazone, cholecalciferol, Cilostazol, cimetidine, cinnarizine, cinoxacin, ciprofibrate, ciprofloxacin hydrochloride, cisapride, cisplatin, citalopram, cladribine, clarithromycin, clemastine fumarate, clioquinol, clobazam, clofarabine, clophenazine, clofibrate, clomiphene, clomipramine, clonazepam, clopidogrel, clotiazepam, clotrimazole, cloxacillin, clozapine, cocaine, codeine, colchicine, colistin, conjugated estrogens, corticosterone, cortisone, cortisone acetate, cyclizine, cyclohexylbarbital, cyclobenzaprine, cyclobutane-spirobarbiturate, cyclohexane-spirobarbiturate, cycloheptane-spirobarbiturate, cyclohexane-spirobarbiturate, cyclopentane-spirobarbiturate, cyclophosphamide, cyclopropane-spirobarbiturate biturates, cycloserine, cyclosporine, cyproheptadine, cyproheptadine hydrochloride, cytarabine, cytosine, dacarbazine, dactinomycin, danazol, dananthrone, dantrolene sodium, dapsone, darbepoetin alfa, darodipine, daunorubicin, decoquinate, dehydroepiandrosterone, delavirdine, demeclocycline, denileukin, deoxycorticosterone, desoxymethasone, dexamethasone, dextrophenylalanine Amine, dexchlorpheniramine, dexfenfluramine, dexrazoxane, dextropropoxyphene, heroin, diatrizoate, diazepam, diazoxide, dichlorophen, dipropionic acid, diclofenac, dicoumarol, didanosine, diflunisal, digitoxin, digoxin, dihydrocodeine, dihydroequilin, dihydroergotamine mesylate, diiodoquinol, diltiazem hydrochloride, diloxanide furoate, dimenhydrinate, dimethoate Moramine, dinitolamide, diosgenin, diphenoxylate hydrochloride, biphenyl, dipyridamole, dirithromycin, disopyramide, disulfiram, diuron, docetaxel, domperidone, donepezil, doxazosin, doxazosin hydrochloride, doxorubicin (neutral), doxorubicin hydrochloride, doxycycline, drotasteride propionate, droperidol, dihydroxyprophylline, echinocandins, econazole, econazole nitrate, efavirenz, ellipticine, enalapril, enmosumab, enoximone, epinephrine, epipodophyllotoxin derivatives, epirubicin, epoetin alfa, ceftazidime, dehydroequilin, equilin, ergocalciferol, ergotamine tartrate, erlotinib, erythromycin, estradiol, estramustine, estriol, estrone, ethacrynic acid, ethambutol, ethinyl amine , ethionamide, profenamide hydrochloride, ethyl 4-aminobenzoate (benzocaine), ethyl hydroxybenzoate, ethinyl estradiol, etodolac, etomidate, etoposide, etretinate, exemestane, felbamate, felodipine, fenbendazole, fenbufen, fenclofos, fencloric acid, fenfluramine, fenofibrate, fenoldopam, fenoprofen, fenoxycarb, fenpicrol, fentanyl, fenticonazole, fexofenadine, filgrastim, finasteride, flecainide acetate, floxuridine, fludarabine, fluconazole, fluconazole, flucytosine, fludioxonil, fludrocortisone, fludrocortisone acetate, flufenamic acid, flunarizine, flunarizine hydrochloride, flunisolide, flunitrazepam, fluocortolone, flumethuron, fluorene, fluorouracil,Fluoxetine hydrochloride, fluoxymesterone, triflutixol decanoate, flupentixol decanoate, flurazepam, flurbiprofen, fluticasone propionate, fluvastatin, folic acid, fosinopril, fosphenytoin sodium, frovatriptan, furosemide, fulvestrant, furazolidone, gabapentin, G-BHC (lindane), gefitinib, gemcitabine, gemfibrozil, gemtuzumab, glafenine, glibenclamide, gliclazide, glimepiride, glipizide, glutethimide, glibenclamide, glyceryl trinitrate (nitroglycerin), goserelin acetate, grepafloxacin, griseofulvin, guaifenesin, guanabenzyl acetate, guanine, haloperidol hydrochloride, hydrochlorothiazide, heptylbarbital, heroin, hesperetin, hexachlorobenzene, hexylbarbital, histrelin acetate, hydrocortisone , hydroflumethiazide, hydroxyurea, scopolamine, hypoxanthine, ibritumomab tiuxetan, ibuprofen, idarubicin, allyl butalbital, ifosfamide, ihydroequilenin, imatinib mesylate, imipenem, indapamide, indinavir, indomethacin, indoprofen, interferon alfa-2a, interferon alfa-2b, iodamide, iopanoic acid, iprodione, irbesartan, irinotecan, isavuconazole, isocarboxazid, isoconazole, isoguanine, isoniazid, isopropyl barbiturate, isoproturon, isosorbide dinitrate, isosorbide mononitrate, isradipine, itraconazole, itraconazole, itraconazole (Itra), ivermectin, ketoconazole, ketoprofen, ketorolac, kaline, labetalol, lamivudine, lamotrigine, tricholoma Azapropazone, lansoprazole, L-DOPA, leflunomide, lenalidomide, letrozole, leucovorin, leuprolide, levamisole, levofloxacin, lidocaine, linduron, lisinopril, lomefloxacin, lomustine, loperamide, loratadine, lorazepam, lorefloxacin, clometazepam, losartan mesylate, lovastatin, lisuride maleate, maprotiline hydrochloride, clomiphene citrate, mebendazole, meclizine hydrochloride, meclofenamic acid, medazepam, medigoxin, medroxyprogesterone acetate, mefenamic acid, mefloquine hydrochloride, megestrol acetate, melphalan, mepenzolate bromide, meprobamate, meptazocine, mercaptopurine, mesalamine, mesna, meso pyridazine, mestranol, methadone, methaqualone, methocarbamol, mephenytoin, methotrexate, methoxsalen, methsuximide, methylclothiazide, methylphenidate, methiothiazide, methyl-paraben, methylprednisolone, methyltestosterone, methysergide, metoclopramide, metolazone, metoprolol, metronidazole, mianserin hydrochloride, miconazole, midazolam, mifepristone, miglitol, minocycline, minoxidil, mitomycin C, mitotane, mitoxantrone, mycophenolate mofetil, molindone, montelukast, morphine, moxifloxacin hydrochloride, nabumetone, nadolol, nalbuphine, nalidixic acid, nandrolone, naphthane, naphthalene, naproxen, naratriptan hydrochloride, natamycin, nelarabine, nelfinavir, nevirapine, nicardipine hydrochloride, nicotinamide, niacin,Acenocoumarol, nifedipine, nilutamide, nimodipine, nimozole, nisoldipine, nitrazepam, nitrofurantoin, nitrofurazone, nizatidine, nomostat, norethisterone, norfloxacin, norgestrel, nortriptyline hydrochloride, nystatin, estradiol, ofloxacin, olanzapine, omeprazole, omoconazole, ondansetron hydrochloride, oprelekin, ornidazole, oxaliplatin Platinum, oxaniquine, oxaprozin, oxamit, oxazepam, oxcarbazepine, oxfendazole, oxiconazole, oxprenolol, hydroxybutazone, hydroxybenzylamine hydrochloride, paclitaxel, palifermin, pamidronate, para-aminosalicylic acid, pantoprazole, mefenamic acid, paroxetine hydrochloride, pegaspargase, pegfilgrastim, pemetrexed disodium, penicillamine , pentaerythritol nitrate, pentazocine, pentazocine, pentobarbital, pentobarbital, pentostatin, pentoxifylline, perphenazine, perphenazine pimozide, perylene, phenylacetylurea, phenacetin, phenanthren, phenindione, phenobarbital, barbiturate, phenolphthalein, phenoxybenzamine, phenoxybenzamine hydrochloride, phenoxymethylpenicillin, phensuximide, phenylbutazone, phenytoin, pindolol, pioglitazone, pipobroman, piroxicam, pizothiazine maleate, platinum compounds, plicamycin, polyenes, polymyxin B, porfimer sodium, posaconazole (Posa), pramipexole, prasterone, pravastatin, praziquantel, prazosin, prazosin hydrochloride, prednisolone, prednisone, primidone, probarbital, probenecid, probucol, procarbazine, prochlorperazine, progesterone, chlorpromazine hydrochloride Guanidine, promethazine, propofol, propoxur, propranolol, propylparaben, propylthiouracil, prostaglandins, pseudoephedrine, pteridine-2-methylthiol, pteridine-2-thiol, pteridine-4-methylthiol, pteridine-4-thiol, pteridine-7-methyl 1-thiol, pteridine-7-thiol, pyrantel pamoate, pyrazinamide, pyrene, pyridostigmine, pyrimethamine, quetiapine, mipaline, quinapril, quinidine, quinidine sulfate, quinine, quinine sulfate, rabeprazole sodium, ranitidine hydrochloride, rasburicase, ravuconazole, repaglinide, dicyclopental, reserpine, retinoids, rifabutin, rifampicin, rifapentine, rimexolone, risperidone, ritonavir, rituximab, rizatriptan, benzoate, rofecoxib, Ropinirole hydrochloride, rosiglitazone, saccharin, salbutamol, salicylamide, salicylic acid, saquinavir, sargramostim, butabarbital, secobarbital, sertaconazole, sertindole, sertraline hydrochloride, simvastatin, sirolimus, sorafenib, sparfloxacin, spiramycin, spironolactone, dihydrotestosterone, stanozolol, stavudine, diethylstilbestrol, streptozocin, strychnine, sulconazole, sulconazole nitrate, sulfacetamide, sulfadiazine, sulfadiazine, sulfadimethoxine, sulfamethoxazole, sulfa, sulfathiazole, sulindac, sulfonamide, sulfacetamide, sulfadiazine, sulfadoxine, sulfisoxazole, sulfamethoxazole, sulfapyridine, sulfasalazine, sulfinpyrazone, sulpiride, sultimethoxazole, sumatriptan succinate,sunitinib maleate, tacrine, tacrolimus, talbutal, tamoxifen citrate, tamsulosin, bexarotene, taxanes, tazarotene, telmisartan, temazepam, temozolomide, teniposide, tenoxicam, terazosin, terazosin hydrochloride, terbinafine hydrochloride, terbutaline sulfate, terconazole, terfenadine, testolactone, testosterone, tetracycline, tetrahydrocannabinol, tetraoxoprene, thalidomide, thebaine, theobromine, theophylline, thiabendazole, thiamphenicol, thioguanine, thioridazine, thiotepa, thotoin, thymine, tiagabine hydrochloride, tibolone, ticlopidine, tinidazole, tioconazole, tirofiban, tizanidine hydrochloride, tolazamide, tolbutamide, tolcapone , topiramate, topotecan, toremifene, tositumomab, tramadol, trastuzumab, trazodone hydrochloride, tretinoin, triamcinolone, triamterene, triazolam, triazoles, triflupromazine, trimethoprim, trimipramine maleate, triphenylene, troglitazone, tromethamine, tropicamide, trovafloxacin, tabamate, ubidecarenone (coenzyme Q10), undecylenic acid, uracil, uracil mustard, uric acid, valproic acid, valrubicin, valsartan, vancomycin, venlafaxine hydrochloride, vigabatrin, pentobarbital, vinblastine, vincristine, vinorelbine, voriconazole, xanthine, zafirlukast, zidovudine, zileuton, zoledronate, zoledronic acid, zolmitriptan, zolpidem and zopiclone. ,

[0056] In certain aspects, the therapeutic agent may be voriconazole or other members of the general class of azole compounds. Exemplary antifungal azoles include: a) imidazoles such as miconazole, ketoconazole, clotrimazole, econazole, omoconazole, bifonazole, butoconazole, fenticonazole, isoconazole, oxiconazole, sertaconazole, sulconazole, and tioconazole; b) triazoles such as fluconazole, itraconazole, isavuconazole, ravuconazole, posaconazole, voriconazole, terconazole; and c) thiazoles such as abafungin. Other drugs that may be used in the method include, but are not limited to, hyperthyroidism drugs such as thyroxine; anticancer agents such as cytotoxic agents such as epipodophyllotoxin derivatives, taxanes, bleomycins, anthracyclines, and platinum compounds and camptothecin analogs. The following therapeutic agents may also include other antifungal antibiotics such as poorly soluble echinocandins, polyenes (e.g., amphotericin B and natamycin) and antibacterial agents (e.g., polymyxin B and colistin) and antiviral drugs. Therapeutic agents may also include therapeutic agents for mental illnesses such as antipsychotics, antidepressants or analgesics and / or tranquilizers such as benzodiazepines. Therapeutic agents also include agents for altering the level of consciousness or anesthetics, such as propofol. The present composition and its preparation method can be used to prepare pharmaceutical compositions having appropriate pharmacokinetic properties for use as therapeutic drugs.

[0057] In some embodiments, the compositions described herein may include a long-acting beta-agonist (LABA). Some non-limiting examples of long-acting beta-agonists include formoterol such as formoterol fumarate, salmeterol such as salmeterol xinafoate, bambuterol, clenbuterol hydrochloride, indacaterol, olopataterol, protocol, abedeterol, salmethamine, vilanterol, arformoterol, carmoterol, SK-PF-610355, GSK-159797, GSK-597901, GSK-159802, GSK-642444, GSK-678007 or other long-acting beta-agonists known in the art.

[0058] In other embodiments, the compositions described herein may include a long-acting muscarinic antagonist (LAMA). Some non-limiting examples of long-acting muscarinic antagonists include tiotropium, aclidinium, daptomycin, ipratropium, oxitropium, daptomycin, glycopyrrolate or glycopyrrolate derivatives or other long-acting muscarinic antagonists consistent in the art such as those taught in U.S. Patent Application No. 2009 / 0181935, PCT Patent Application No. WO 2010 / 007561, and PCT Patent Application No. WO 2008 / 035157, which are incorporated herein by reference.

[0059] In other embodiments, the compositions described herein may include a corticosteroid, particularly a corticosteroid suitable for inhalation. Some non-limiting examples of corticosteroids include beclomethasone dipropionate, budesonide, flunisolone, fluticasone propionate, fluticasone furoate, mometasone furoate, ciclesonide, roflufenolone palmitate, triamcinolone acetonide, or other corticosteroids known in the art.

[0060] In other embodiments, the compositions described herein may include one or more antibiotics. Some antibiotic classes include penicillins, cephalosporins, carbapenems, macrolides, aminoglycosides, quinolones (including fluoroquinolones), sulfonamides and tetracyclines. In certain embodiments, the compositions may include narrow-spectrum antibiotics for specific bacterial types. In the non-limiting examples of some antibiotics, penicillins, cephalosporins, polymyxins, rifamycins, ribimycins, quinolones and sulfonamides are included. In some non-limiting examples of antibacterial antibiotics, macrolides, lincosamides or tetracyclines are included. In some embodiments, the antibiotic is: an aminoglycoside such as kanamycin and streptomycin; ansamycins such as rifaximin and geldanamycin; carbenicillins such as loracarbef; carbapenems such as ertapenem, imipenem; cephalosporins such as cephalexin, cefixime, cefepime and ceftobiprole; glycopeptides such as vancomycin or teicoplanin; lincosamides such as lincomycin and clindamycin; lipopeptides such as daptomycin; macrolides such as clarithromycin, spiramycin, azithromycin and telithromycin; monoclonal antibodies such as chloramphenicol, ... Bacillus amides, such as aztreonam; nitrofurans, such as furazolidone and nitrofurantoin; oxazolidinones, such as linezolid; penicillins, such as amoxicillin, azlocillin, flucloxacillin and penicillin G; antibiotic polypeptides, such as bacitracin, polymyxin B and colistin; quinolones, such as ciprofloxacin, levofloxacin and gatifloxacin; sulfonamides, such as silver sulfadiazine, mafenide, sulfadimethoxine or sulfapyridine; or tetracyclines, such as demeclocycline, doxycycline, minocycline, oxytetracycline or tetracycline. In some embodiments, the composition comprises drugs that act on mycobacteria, such as cycloserine, capreomycin, ethionamide, rifampicin, rifabutin, rifapentine and streptomycin. Other antibiotics that are contemplated include arsphenamine, chloramphenicol, fosfomycin, fusidic acid, metronidazole, mupirocin, platenmycin, quinupristin, dalfopristin, thiamphenicol, tigecycline, metronidazole or trimethoprim.

[0061] In some embodiments, the composition may further comprise one or more antifungal agents, such as those described above. Some antifungal agents include, but are not limited to, amphotericin B (an azole antifungal compound), echinocandins or flucytosine. Some non-limiting examples of azole antifungal compounds include: a) imidazoles, such as miconazole, ketoconazole, clotrimazole, econazole, dimethoconazole, bifonazole, butoconazole, fenticonazole, isoconazole, oxiconazole, sertaconazole, sulconazole and tioconazole; b) triazoles, such as fluconazole, itraconazole, isavuconazole, ravuconazole, posaconazole, voriconazole, terconazole; and c) thiazoles, such as abafungin.

[0062] In some embodiments, the composition may further comprise one or more antiviral agents such as nucleosides, for example, acyclovir, famciclovir, valacyclovir, penciclovir, and ganciclovir, or other antiviral agents such as pegylated interferon, interferon alpha-2b, lamivudine, adefovir, telbivudine, entecavir, or tenofovir.

[0063] B. Excipients

[0064] In some aspects, the disclosure includes one or more excipients formulated into a pharmaceutical composition. In certain embodiments, the excipient used herein is a water-soluble excipient. These water-soluble excipients include carbohydrates, such as disaccharides, such as sucrose, trehalose or lactose; trisaccharides, such as fructose, glucose, galactose or raffinose; polysaccharides, such as starch or cellulose; or sugar alcohols, such as xylitol, sorbitol or mannitol. In certain embodiments, these excipients are solid at room temperature. Some non-limiting examples of sugar alcohols include erythritol, threitol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fructose, iditol, inositol, heptyl alcohol, isomalt, maltitol, lactitol, maltotriitol, maltotetraitol or polyglucitol. In certain aspects, the pharmaceutical composition of the present invention may further be free of hydrophobic or wax excipients such as wax and oil. Some non-limiting examples of hydrophobic excipients include hydrogenated oil and partially hydrogenated oil, palm oil, soybean oil, castor oil, carnauba wax, beeswax, palm wax, white wax, castor wax or lanolin. In addition, the disclosure may further include one or more amino acids or their amides or ester derivatives. In certain embodiments, the amino acid used can be one of 20 kinds of classical amino acids, such as glycine, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, tryptophan, serine, threonine, asparagine, glutamine, cysteine, selenocysteine, proline, arginine, histidine, lysine, aspartic acid or glutamic acid. These amino acids can be D or L orientation, or can be alpha-amino acid, beta-amino acid, gamma-amino acid or delta-amino acid. In other embodiments, one of common non-classical amino acids can be used, such as carnitine, GABA, carboxyglutamic acid, levothyroxine, hydroxyproline, selenomethionine, beta-alanine, ornithine, citrulline, dehydroalanine, delta-aminolevulinic acid or 2-aminoisobutyric acid.

[0065] In some aspects, the amount of excipient in the pharmaceutical composition is about 0.5% w / w to about 10% w / w, about 1% w / w to about 10% w / w, about 2% w / w to about 8% w / w, or about 2% w / w to about 5% w / w. The amount of excipient in the pharmaceutical composition is about 0.5% w / w, 0.75% w / w, 1% w / w, 1.25% w / w, 1.5% w / w, 1.75% w / w, 2% w / w, 2.5% w / w, 3% w / w, 3.5% w / w, 4% w / w, 4.5% w / w, 5% w / w, 6% w / w, 7% w / w, 8% w / w, 9% w / w, to about 10% w / w, or any range derivable therein, based on the total weight of the pharmaceutical composition. In one embodiment, the amount of excipient in the pharmaceutical composition is 2% w / w to 5% w / w of the total weight of the pharmaceutical composition.

[0066] II. Manufacturing Method

[0067] Therefore, in one aspect, the present disclosure provides a pharmaceutical composition that can be prepared using a thin film freezing method. Such methods are described in U.S. Patent Application No. 2010 / 0221343 and Watts et al. (2013), which are incorporated herein by reference. In some embodiments, these methods include dissolving the ingredients of the pharmaceutical composition in a solvent to form a precursor solution. The solvent can be water or an organic solvent. Some non-limiting examples of organic solvents that can be used include volatile organic solvents, such as 1,4-dioxane, acetonitrile, acetone, methanol, ethanol, isopropanol, dichloromethane, chloroform, tetrahydrofuran, tert-butyl alcohol, dimethyl sulfoxide, N, N-dimethylformamide, ether, ethyl acetate, isopropyl acetate, butyl acetate, propyl acetate, toluene, hexane, heptane, pentane or a combination thereof. In some embodiments, the precursor solution may include less than 10% w / v of the therapeutic agent and excipient. The precursor solution may comprise less than 0.5% w / v, 1% w / v, 2% w / v, 3% w / v, 4% w / v, 5% w / v, 6% w / v, 7% w / v, 8% w / v, 9% w / v or 10% w / v of the therapeutic agent and excipient, or any range derived therefrom.

[0068] The precursor solution may be deposited onto a surface at a temperature that causes the precursor solution to freeze. In some embodiments, the temperature may be below the freezing point of the solution at ambient pressure. In other embodiments, a reduced pressure may be applied to the surface to freeze the solution at a temperature below the freezing point at ambient pressure. The surface may also be rotated or moved on a moving conveyor system so that the precursor solution is evenly distributed on the surface. Alternatively, the precursor solution may be applied to the surface in a manner that forms a flat surface.

[0069] After the precursor solution is applied to the surface, the solvent can be removed to obtain a pharmaceutical composition. Any suitable method of removing the solvent can be adopted, including evaporation under reduced pressure or high temperature or freeze-drying conditions. In certain embodiments, freeze-drying may include reduced pressure and / or reduced temperature. Such reduced temperature can be 25°C to about -200°C, 20°C to about -175°C, about 20°C to about -150°C, 0°C to about -125°C, -20°C to about -100°C, -75°C to about -175°C or -100°C to about -160°C. The temperature is about -20°C, -30°C, -35°C, -40°C, -45°C, -50°C, -55°C, -60°C, -70°C, -80°C, -90°C, -100°C, -110°C, -120°C, -130°C, -140°C, -150°C, -160°C, -170°C, -180°C, -190°C, to about -200°C or any range derivative therein. Additionally, the solvent may be removed at a reduced pressure of less than 500 mTorr, 450 mTorr, 400 mTorr, 375 mTorr, 350 mTorr, 325 mTorr, 300 mTorr, 275 mTorr, 250 mTorr, 225 mTorr, 200 mTorr, 175 mTorr, 150 mTorr, 125 mTorr, 100 mTorr, 75 mTorr, 50 mTorr, or 25 mTorr.

[0070] The compositions prepared using these methods may exhibit brittleness, making the compositions easy to shear into smaller particles when processed by the equipment. These compositions have a high surface area and exhibit improved fluidity of the composition. Such fluidity can be measured by, for example, the Carr index or other similar indicators. In particular, the Carr index can be measured by comparing the bulk density of the powder with the tap density of the powder. When the composition is processed by an auxiliary device to deliver the drug, such compounds may exhibit a favorable Carr index and may result in better shearing of the particles to produce smaller particles.

[0071] III. Definitions

[0072] When used with the term "comprising" in the claims and / or the specification, the use of the word "a" can mean "one", but it is also consistent with the meaning of "one or more", "at least one", and "one or more than one". As used herein, "another" can mean at least a second or more.

[0073] As used herein, the terms "drug", "medication", "therapeutic agent" and "therapeutically active agent" are used interchangeably to refer to compounds that cause a therapeutic or pharmacological effect in humans or animals and are used to treat a disease, disorder or other condition. In some embodiments, these compounds have been and have received regulatory approval for administration to living animals.

[0074] The use of the term "or" in the claims means "and / or" unless explicitly stated to refer to only alternatives or the alternatives are mutually exclusive. As used herein, "another" may mean at least a second or more.

[0075] As used in this specification and claims, the words "comprising" (and any form of "comprising", such as "comprise" and "comprises"), "having" (and any form of "having", such as "have" and "has"), "including" (and any form of "including", such as "includes" and "include"), or "containing" (and any form of "containing", such as "contains" and "contain") are non-exhaustive or open-ended and do not exclude additional, unrecited elements or method steps.

[0076] As used in this specification, the term "significant" (and any form of "significant," such as "significantly") is not intended to imply a statistical difference between two values, but only to imply the significance of a parameter or the extent of the difference.

[0077] Throughout this application, the term "about" is used to indicate that a value includes the inherent error variation of the device, method employed to determine the value, or the variation that exists between research subjects or experimental studies. Unless otherwise defined, the term "about" means ± 10% of the stated value.

[0078] As used herein, with respect to a particular component, the term "substantially free" or "substantially free" is used herein to mean that the particular component is not intentionally formulated into the composition and / or the particular component is present only as a contaminant or in trace amounts. The total amount of all containers, byproducts, and other materials present in the composition is less than 2%. The term "substantially free" or "substantially free" means that the composition contains less than 1% of the particular component. The term "substantially free" or "substantially free" contains less than 0.5% of the particular component.

[0079] As used herein, the term "domain" refers to a specific region of a composition that is substantially composed of a single material that has characteristics that are different from other components of the composition. A "discrete domain" refers to a separate region of a composition that is distinct from one another and separate from other regions of the composition. The domain may be substantially composed of a single element in the composition. The domains may be discontinuous, such that the discrete domains exist as multiple domains that are not in contact with one another.

[0080] As used herein, the term "nanoparticle" has its conventional and ordinary definition and refers to a discrete particle that behaves as an integral unit rather than as a single molecule within the particle. Nanoparticles may have a size of about 1 nm to about 10,000 nm, ultrafine nanoparticles have a size of 1 nm to 100 nm, fine particles have a size of 100 nm to 2,500 nm, and coarse particles have a size of 2,500 nm to 10,000 nm. In some embodiments, the nanoaggregates described herein may comprise a combination of a plurality of nanoparticles and have a size of about 10 nm to about 100 μm.

[0081] Although the numerical ranges and parameters set forth within the broad scope of the invention are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as possible. Any numerical values, however, inherently contain certain errors necessarily resulting from the standard deviation found in their respective measurements and parameters.

[0082] Through the following detailed description, other purposes, features and advantages of the present disclosure will become apparent. However, it should be understood that the specific embodiments and specific examples are only used for demonstration when indicating specific embodiments of the present disclosure, because according to this specific embodiment, various changes and improvements within the spirit and scope of the present disclosure will be apparent to those skilled in the art.

[0083] IV. Examples

[0084] For the convenience of better understanding of the present disclosure, the examples of the following specific embodiments are provided. It will be appreciated by those skilled in the art that the technology disclosed in the following examples represents the technology that the inventor has found to play a good role in the practice of the present disclosure, and therefore it can be considered to constitute the preferred mode of its practice. However, according to the present disclosure, it will be appreciated by those skilled in the art that many changes can be made to the disclosed specific embodiments without departing from the spirit and scope of the present disclosure, and the same or similar results can still be obtained. The following examples should not be construed as limiting or defining the full scope of the present disclosure.

[0085] Example 1 - Discussion and Results

[0086] A. Physicochemical properties of voriconazole dry powder preparations

[0087] Crystalline voriconazole powder formulations containing mannitol were produced using TFF technology (see Table 1). XRPD and mDSC were mainly used to determine the crystallinity of the formulations. The TFF-VCZ powder formulation containing mannitol was identified as crystalline, such as Figure 1 and 2 The XRPD of the TFF-VCZ-MAN powder formulation showed characteristic peaks of voriconazole, corresponding to voriconazole bulk powder (e.g., 12.4°2θ and 13.6°2θ) and δ-mannitol (e.g., 9.5°2θ and 20.2°2θ), as shown in FIG. Figure 1 As shown. These results indicate that the powder formulation consists of crystalline voriconazole and δ-mannitol. In the TFF-VCZ-MAN powder formulation, the peak intensity of δ-mannitol decreases as the mannitol content (% w / w) decreases, and when the powder formulation contains 5% (w / w) mannitol, the peak corresponding to δ-mannitol cannot be detected. TFF-MAN dry powder is mainly δ-form, while XRPD detects trace amounts of α-form and β-form (13.5°2θ and 14.5°2θ, respectively).

[0088] Table 1. Summary of voriconazole dry powder formulations studied using thin film freezing (TFF) technology.

[0089]

[0090]

[0091] mDSC also confirmed the crystallinity of the TFF-VCZ-MAN powder formulation. Figure 2It is shown that no glass transition is detected in TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50, and only endothermic peaks corresponding to the melting of voriconazole and mannitol are detected. TFF-VCZ has a melting endothermic peak at 130.86°C, and its melting heat is 105.3 J / g. When the expected melting heat of voriconazole in TFF-VCZ-MAN powder is calculated by percentage fraction (w / w), the melting heat of voriconazole in TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50 is 100.0 J / g and 52.6 J / g, respectively. The measured melting heat of voriconazole in TFF-VCZ-MAN 95:5 is 95.1 J / g, and the measured melting heat of voriconazole in TFF-VCZ-MAN 50:50 is 33.7 J / g, which are 95.1% and 64.0% of the expected values, respectively. TFF-MAN has a melting endotherm at 167.31°C with a melting heat of 187.5 J / g. The expected melting heats of mannitol in TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50 are 9.38 J / g and 93.8 J / g, respectively. The measured melting heats of mannitol are 2.63 J / g and 63.2 J / g, respectively, and are 28.0% and 67.4% of the expected values, respectively. Table 2 lists the composition ratios of voriconazole to mannitol (voriconazole: mannitol, w / w) in the two formulations measured by mDSC. 1 H-NMR calculates these ratios by integrating the proton peaks. The theoretical ratio for one proton of TFF-VCZ-MAN 95:5 is 1:0.1009, and the calculated experimental ratio is 1:0.0992, indicating that 98.3% of the expected mannitol was detected in TFF-VCZ-MAN 95:5. In the case of TFF-VCZ-MAN 50:50, 100% of the expected mannitol was detected using 1H-NMR.

[0092] Table 2. Utilization 1 Quantitative comparison of voriconazole and mannitol in TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50 by H-NMR.

[0093]

[0094]

[0095] The particle morphology of the TFF-VCZ-MAN powder is shown in Figure 3. Agglomeration of micron-sized particles was observed in the TFF-VCZ powder, and these particles were also found in other TFF-VCZ-MAN powder formulations. A more porous matrix was observed in the TFF-VCZ-MAN powder containing a higher content of mannitol. The 3D morphology and illustration of the TFF-VCZ and TFF-VCZ-MAN 95:5 powders shown in Figure 4 confirmed that the surface texture of the TFF-VCZ-MAN 95:5 powder was rough, while the surface texture of the TFF-VCZ powder was smooth. Figure 5 The high-resolution morphology of TFF-VCZ-MAN 95:5 powders in Figure 2 shows that TFF-VCZ-MAN 95:5 powders are nanoaggregates composed of nanoparticles with an average size of about 150-500 nm. The SSA of these TFF-VCZ-MAN powders is shown in Figure 2. Figure 6 As shown in Figure 2, TFF-VCZ powder exhibited the lowest SSA (8.36 m 2 / g), while the porous matrix of TFF-MAN dry powder showed the highest SSA (17.11m 2 / g). SSA increases with the addition of mannitol in the TFF-VCZ-MAN powder formulation. With the help of SEM / EDX shown in Figure 7, the micron-sized particles are identified as being composed of voriconazole nanoaggregates by the detection results of nitrogen, oxygen and fluorine. The porous matrix is ​​identified as mannitol by the detection results of oxygen (without nitrogen and fluorine).

[0096] The FT-IR peak pattern of TFF-VCZ powder matched that of voriconazole bulk powder, and it was also found that TFF-VCZ-MAN powders containing different amounts of mannitol had the same peak pattern. The peak pattern of TFF-MAN was also found in TFF-VCZ-MAN powder. Therefore, only peaks corresponding to TFF-VCZ and TFF-MAN were observed in TFF-VCZ-MAN powder, and no new peaks were found in the FT-IR spectrum of TFF-VCZ-MAN powder, as shown in Figure 8. 1D spectroscopy obtained by ssNMR 13 C and 19 The F CP-MAS spectrum is shown in Figure 9. Voriconazole and mannitol do not exist 13 The spectra of the C peaks overlap and 19 F spectrum has all resonances. TFF-VCZ and TFF-VCZ-MAN show the same spectrum. In addition, the TFF-VCZ-MAN 90:10 spectrum 13 C and 19 The F peak confirms the crystallinity of voriconazole and mannitol. 2D of TFF-VCZ-MAN 90:10 1 H- 13Comparison of the CHETCOR spectrum with that of TFF-VCZ is shown in Figure 10. No intermolecular cross peaks between voriconazole and mannitol from TFF-VCZ-MAN 90:10 were observed.

[0097] B. In vitro aerosol performance and stability

[0098] The aerodynamic particle size distribution of TFF-VCZ-MAN powder formulation was determined using NGI, and the FPF (quantitative dose percentage) was as follows: Fig.11 As shown. Based on the FPF (quantitative dose percentage) data, the TFF-VCZ-MAN powder formulation composed of 90% to 97% (w / w) voriconazole showed the highest aerosol effect. The FPF (quantitative dose percentage) of TFF-VCZ-MAN 97:3 was significantly higher (p<0.05) than that of TFF-VCZ, and the FPF (quantitative dose percentage) was improved by 66%. There was no significant difference in the aerosol performance of TFF-VCZ-MAN powder containing 90% to 97% (w / w) voriconazole (p>0.05). When the composition contains more than 10% (w / w) mannitol, the aerosol performance of the TFF-VCZ-MAN powder formulation decreases.

[0099] The effect of physical forces on the aerosol performance of the TFF-VCZ-MAN 95:5 powder formulation was also investigated by measuring FPF using NGF. Fig.12 As shown in Figures 1 and 13, the change of particle size distribution and aerosol performance was monitored at different shearing times. At 15min, 30min and 60min, FPF (quantitative dose percentage) was 44.3%, 47.5% and 42.4% respectively, and FPF (delivered dose percentage) was 68.7%, 73.6% and 69.5% respectively. Before applying shear force, the initial value of FPF was 40.0% (quantitative dose percentage), and the initial value of FPF was 58.8% (delivered dose percentage). Although it is also observed that MMAD changes from 3.7 μm at the initial time to 3.2 μm, 3.0 μm and 3.1 μm at 15min, 30min and 60min, GSD does not see significant changes.

[0100] A stability study was conducted at 25°C / 60%RH and the purity and aerosol performance of the TFF-VCZ-MAN 95:5 powder formulation were monitored over 13 months, and the results are shown in Figure 14. During the test period, the purity of voriconazole in TFF-VCZ-MAN 95:5 remained unchanged, and no degradation products were detected. In the stability study, FPF (percentage of quantitative dose), FPF (percentage of delivered dose), MMAD, and GSD were compared at each time point to compare aerosol performance. Within 13 months, there were no statistically significant differences in FPF (percentage of quantitative dose) and FPF (percentage of delivered dose) (both p values ​​were greater than 0.05). After 1 month, GSD decreased significantly from the initial value (p<0.05), while MMAD did not show any difference within 13 months (p>0.05).

[0101] C. Discussion of voriconazole dry powder formulation

[0102] In the dissolution test of TFF-VCZ-MAN powder formulation, pH 7.4 PBS was used as the receiving medium, and the top of the Franz cell supply chamber was covered with paraffin film to prevent evaporation loss of the solvent. The dissolution of crystalline TFF-VCZ-MAN95:5 was compared with that of amorphous TFF-VCZ-PVPK25 25:75. The results showed that the cumulative drug release rate of the crystalline dry powder during the test period was significantly lower (p<0.05), as shown in Table 1. Fig.15 The cumulative voriconazole release rate of amorphous TFF-VCZ-PVPK25 within 3 hours was 63.2% w / v, while the cumulative voriconazole release rate of crystalline TFF-VCZ-MAN 95:5 was only 22.8%. The cumulative voriconazole release rates of TFF-VCZ-MAN 25:75 and TFF-VCZ-MAN 50:50 within 3 hours were 46.3% and 35.3%, respectively.

[0103] D. Characterization of voriconazole dry powder formulation

[0104] Voriconazole (Beinborn et al., 2012b; Ramos and Diogo, 2016) and mannitol (Yu et al., 1998) have a high tendency to crystallize and have glass transition temperatures below room temperature. Therefore, unless there is a strong intermolecular interaction between voriconazole and mannitol to prevent crystallization, it is assumed that TFF-VCZ-MAN is in a crystalline state. Based on the XRPD data and the sharpness of the 1D CP-MAS spectrum, the TFF-VCZ-MAN powder formulation is in a crystalline state, indicating that there is no sufficiently strong interaction between voriconazole and mannitol.

[0105] While XRPD can be used to characterize the crystallinity of a powder, it may not be able to detect small amounts of amorphous forms in the formulation. Therefore, mDSC analysis of the TFF-VCZ-MAN powder was performed, and the results indicated that the TFF-VCZ-MAN dry powder was in a crystalline state as only two endothermic peaks for voriconazole and mannitol were detected. However, a decrease in the melting point of mannitol was observed, especially in TFF-VCZ-MAN 95:5. The heat of fusion of mannitol in TFF-VCZ-MAN 95:5 may be lower because a relatively small amount of mannitol is dissolved in molten voriconazole before the temperature reaches the melting point of mannitol. Additionally, the mannitol particles in TFF-VCZ-MAN 95:5 are typically 100-200 nm, and these nanosized particles can reduce the heat of fusion. To confirm the efficacy of mannitol in TFF-VCZ-MAN powder that exhibits a low melting point, the nanometer-sized particles were used to characterize the crystallinity of the TFF-VCZ-MAN powder. 1 H-NMR was used to determine the molecular ratio between voriconazole and mannitol. Although NMR is usually used for qualitative analysis, quantitative NMR analysis can also be used (Espina et al., 2009; Pauli et al., 2012). In both TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50, the experimental molecular ratio between voriconazole and mannitol was in good agreement with the theoretical value. 13 C and 19 F ssNMR confirms crystalline polymorphism and identifies low levels of amorphous drug in solid dosage forms (Correa-Soto et al., 2017; Offerdahl et al., 2005). 13 C and 19 The peak positions and line widths of the voriconazole resonances in the F CP-MAS spectrum are identical, confirming its crystallinity and indicating the absence of quantifiable amorphous content.

[0106] FTIR was used to study the chemical interaction between voriconazole and mannitol. The hydroxyl group of voriconazole is associated with its degradation pathway (Shaikh and Patil, 2012) and it may be the most active site in the presence of intermolecular interactions. If this occurs, the FT-IR peak of voriconazole will be at 3100 cm -1 With 3500cm -1 In this range, there are two peaks corresponding to voriconazole, which are at 3118.9 cm -1 and 3198.4cm -1 These two peaks were observed in all TFF-VCZ-MAN and TFF-VCZ powder formulations, but no shift was found between the two peaks.-1 The peak of mannitol was observed at 1230 cm-1, and no shift was observed. If there is an interaction between mannitol and the aromatic secondary amine of voriconazole, it may be that the peak at 1230 cm-1 is -1 With 1300cm -1 Peak shifts were observed between 1241.5 cm and 1246.5 cm (Silverstein et al., 2005). -1 、1248.8cm -1 、1268.5cm -1 and 1277.6cm -1 Four peaks of voriconazole were detected at 100 nm, but no obvious peak shift was found when mannitol was included in the voriconazole powder formulation. Therefore, these FT-IR data support that there is no or very weak interaction between voriconazole and mannitol in the TFF-VCZ-MAN powder formulation.

[0107] Although FT-IR is commonly used to identify conformations and intermolecular interactions, ssNMR is able to provide more in-depth atomic-level information for structural studies (Tian et al., 2017). 13 C and 19 F CP-MAS was used to investigate the conformational changes. There was no difference in the chemical shifts of all voriconazole peaks between TFF-VCZ and TFF-VCZ-MAN 90:10. 13 C- 1 H HETCOR spectra in order to study structural perturbations at a higher resolution. This result confirms that there is no 13 C chemical shift changes. At the given resolution, the chemical shifts of all aliphatic and aromatic protons in the indirect dimension also showed no observable changes. 13 C- 1 H HETCOR detects drug-excipient interactions. No intermolecular cross peaks (i.e., interactions) between voriconazole and mannitol were observed at a given spectral intensity.

[0108] Two different shapes of particles were observed in the TFF-VCZ-MAN powder formulation, and based on the observed particle morphology of TFF-VCZ and TFF, it was initially believed that the micron-sized particles were voriconazole and the porous matrix was mannitol. To confirm this, the chemical composition of these particles was confirmed using SEM / EDX. However, during the initial SEM / EDX run, the positions of oxygen, fluorine, and nitrogen detected overlapped with each other, indicating the presence of particles composed of voriconazole and mannitol. The reason was later determined. Since the measurement depth of EDX is in the micron range, the detection beam passes through the entire particle depth of the TFF-VCZ-MAN 50:50 powder being detected. To overcome this problem, the powder was widely dispersed on a carbon tape on a sample holder, and spot analysis was performed to determine the chemical composition of the two different morphologies of particles. Through spot analysis, the micron-sized particles were identified as voriconazole nanoaggregates based on the chemical composition of oxygen, nitrogen, and fluorine, while the porous matrix was identified as mannitol (showing a chemical composition containing oxygen but not nitrogen and fluorine). Therefore, it can be concluded that crystalline mannitol and crystalline voriconazole phase separated during the TFF process.

[0109] Although Figure 5 The AFM images in show that the TFF-VCZ-MAN powder is nanoaggregates, but the BET data also supports the formation of voriconazole nanoaggregates. While the SEM images show that the TFF-VCZ particles are much larger than the highly porous matrix of TFF-MAN, the specific surface area of ​​TFF-MAN is only two times larger than that of TFF-VCZ. This is probably because the voriconazole particles are nanoaggregates with a larger specific surface area than what is visible on the SEM images.

[0110] E. Mannitol content affects aerosol performance and dissolution rate

[0111] The content of mannitol in TFF-VCZ-MAN powder affects its morphology. When a small amount of mannitol is included, the high supercooling that occurs during the TFF process prevents particle growth, thereby forming submicron mannitol particles (Engstrom et al., 2008). These particles are present on the surface of voriconazole nanoaggregates and change their surface texture. During the atomization process, these submicron mannitol particles are not removed from the surface of voriconazole nanoaggregates. This may be due to the difficulty in removing nanoparticles from the surface. Although cohesion and adhesion are proportional to the particle diameter, the removal force (gravity, vibration and centrifugal force) is proportional to the cube of the diameter (Hinds, 1999). Therefore, submicron mannitol particles are difficult to separate from voriconazole nanoaggregates, and the surface of voriconazole nanoaggregates remains rough during the atomization process, thereby making the atomization effect higher. As the mannitol content in TFF-VCZ-MAN powder increases, a larger porous mannitol matrix is ​​produced. These matrices are not only present on the surface of voriconazole nanoaggregate particles, but also surround these nanoaggregate particles. Because the larger porous mannitol matrix holds them together, multiple voriconazole nanoaggregates are assembled. These aggregate structures are retained during the atomization process. Therefore, these large aggregated particles reduce the aerosol performance of TFF-VCZ-MAN powder formulations containing more than 10% (w / w) mannitol.

[0112] The aerosol performance of DPI formulations is closely dependent on the cohesion and adhesion of the particles. These forces include van der Waals forces, surface tension of the adsorbed liquid film, and electrostatic forces (Hickey et al., 1994). All of these factors are affected by particle shape and size, surface roughness / texture, relative humidity, temperature, particle contact duration and speed (Hinds, 1999; Beach et al., 2002; Tan et al., 2016; Price et al., 2002). Among these forces, van der Waals forces are the most important (Hinds 1999). Since van der Waals forces are attractive forces caused by intermolecular dipoles, when the distance between the particle surfaces reaches the separation distance, the van der Waals forces are greatly reduced (Hinds 1999). Therefore, rougher surfaces can significantly reduce van der Waals forces by maintaining a farther average particle distance. Surface roughness affects not only van der Waals forces, but also surface tension caused by surface moisture. Smooth particle surfaces and higher relative humidity lead to stronger surface tension. However, electrostatic forces depend on particle size. Particles larger than 0.1 μm may generate electrostatic forces (Hinds 1999). This attractive electrostatic force is stronger for larger particles and is also related to relative humidity; low humidity retains the charge on the particles for longer. Nevertheless, electrostatic forces are generally considered to be smaller than van der Waals forces and surface tension (Hinds 1999). Therefore, the surface roughness and texture of the particles play an important role in the aerosol performance of DPI formulations.

[0113] The morphological changes of the powder formulations caused by different levels of mannitol significantly affect the aerosol performance of the TFF-VCZ-MAN powder formulations. Changes in the cohesion and adhesion of the particles change the performance of the aerosol, and the reduction of these forces is associated with a reduction in the contact area between the particles (Beach et al., 2002) and a greater distance between the particles (Hinds 1999). The contact area of ​​the TFF-VCZ-MAN nanoaggregates containing a small amount of submicron mannitol particles is significantly reduced, and the distance between the voriconazole particles is further separated, as shown in Figure 4. Compared with TFF-VCZ powder, the FPF (quantitative dose percentage) of TFF-VCZ-MAN 99:1 powder is significantly improved (p<0.05). This improvement caused by the addition of mannitol is present when up to 3% (w / w) of mannitol is added to the formulation. By adding 1% (w / w) mannitol to a formulation containing 97% to 100% (w / w) voriconazole, the FPF (quantitative dose percentage) is increased by about 5%. In addition, the ejection dose of TFF-VCZ-MAN 95:5 powder was approximately 30% higher than that of TFF-VCZ powder (68% and 36%, respectively; data not shown). This enhanced ejection dose was attributed to reduced adhesion of the particles to the device. Since TFF-VCZ-MAN powder produced by TFF contains very little moisture (less than 0.1% w / w; data not shown), and voriconazole and mannitol are not hygroscopic, the surface tension of these particles is expected to be relatively low. Electrostatic forces may be generated by storing powders in low humidity environments, but these forces are thought to be much smaller than van der Waals forces and surface tension (Hinds 1999). Therefore, reducing the contact area and increasing the particle distance by changing the surface texture mainly involves reducing the cohesion and adhesion of the TFF-VCZ-MAN powder formulation, and these reductions in cohesion and adhesion improve the aerosol performance. Young et al. also described a relationship between aerosol performance and the separation energy between particles (Young et al., 2002), which is highly consistent with our results.

[0114] Different contents of mannitol in TFF-VCZ-MAN powder not only affect aerosol performance, but also affect dissolution rate. TFF-VCZ-MAN powder containing higher content of mannitol shows higher dissolution rate, and this result can be explained by faster wetting of powder by mannitol. For TFF-VCZ-MAN powder containing a large amount of mannitol, the surrounding mannitol particles encapsulating voriconazole will wet and dissolve quickly. Therefore, voriconazole nano-aggregates are surrounded by solvent in a short time, and the dissolution rate is accelerated. The SEM image of TFF-VCZ-MAN 25:75 powder shows that on the Franz cell, most of the mannitol particles are dissolved within 5min, while submicron mannitol particles are still observed on the surface of voriconazole nano-aggregates of TFF-VCZ-MAN 95:5 powder. This shows that when only a small amount of mannitol is included in the powder formulation, voriconazole nano-aggregates will not wet quickly.

[0115] F. Benefits of TFF Process

[0116] Highly effective nanoaggregates of voriconazole powder formulations were prepared using TFF. Although carrier-free DPI formulations have been reported previously (Yazdi and Smyth, 2016a; Yazdi and Smyth, 2016b), carriers are often included in DPI formulations. However, carrier-based DPI formulations generally have low efficacy. In addition, there are many factors, such as the particle size (Du et al., 2014), particle size distribution (Steckel and Muller, 1997), and surface morphology (Du et al., 2014; Flament et al., 2004) of the carrier particles, that affect the powder aerosol performance during nebulization, and such factors have a negative impact on the deposited dose uniformity (Du et al., 2017). By using TFF, the highest aerosol performance of TFF-VCZ-MAN nanoaggregates was obtained with as little as 3% (w / w) mannitol; therefore, the efficacy of the optimized TFF-VCZ-MAN powder formulation can be as high as 97% (w / w). This high potency at very low excipient levels allows less powder to be delivered and eliminates the problems of low potency and uneven deposited doses typically caused by carriers.

[0117] Highly effective DPI formulations can also be prepared by other techniques such as grinding. Although the particle size suitable for delivery to the lungs produced by grinding is only a few microns, these particles are still considered as single discrete micron-sized particles. According to the study of Longest and Hindle (2017), voriconazole DPI formulations as nanoaggregates prepared by TFF can have significantly higher total lung absorption efficiency and dose distribution uniformity. It is expected that these voriconazole nanoaggregates can better cover the sites where fungal colonies exist in the epithelium. TFF is able to produce nanoaggregates because it can rapidly nucleate at a freezing rate of up to 10,000K / s, achieve a narrower particle size distribution and a lower Ostwald ripening rate, thereby producing a large number of nuclei and preventing particle growth during freezing (Engstrom et al., 2008; Overhoff et al., 2009). The small size of the unfrozen channels and the rapidly increasing viscosity of the unfrozen solution (Engstrom et al., 2008) make voriconazole nanoaggregates of similar size.

[0118] Surface modification of particles can also be accomplished by TFF. Begat et al. previously reported the use of hydrophobic materials such as lecithin, leucine and magnesium stearate to modify the surface of particles. Although particles treated by dry mechanical fusion methods (e.g., mechanical fusion) show improved aerosol performance with or without a carrier due to the decrease in surface free energy (Begat et al., 2005; Begat et al., 2009), this method is based on the blending of the API with a force control agent (e.g., lecithin, leucine and magnesium stearate). The mechanical fusion method requires the input of mechanical energy into the formulation and may result in increased chemical instability of the drug. In addition, since blending may disaggregate aggregates, surface modification by blending may only be applicable to discrete micron-sized particles, not nano-aggregates. Kawashima et al. also reported surface modification of particles by various methods (such as mechanical shear mixing, freezing or spray drying) (Kawashima et al., 1998). With the help of hydrophilic additives such as light anhydrous silicic acid (AEROSIL), the surface of hydrophobic particles is converted to a hydrophilic surface, and the surface-modified particles have improved in vitro inhalation behavior. However, this method uses discrete micron-sized drug particles and cannot be used for nanoaggregates. Therefore, these discrete micron-sized particles treated by other methods cannot obtain the enhanced absorption and microdosing properties of those nanoaggregates described by Longest and Hindle (2017). However, using TFF, the particle surface can be modified without energy input. The surface of voriconazole nanoaggregates was modified using phase-separated submicron-sized mannitol particles (which were present alone on the surface of the drug nanoaggregates) because their rapid freezing rate prevented particle growth.

[0119] Highly efficient (up to 97% w / w) nanoaggregates of crystalline voriconazole powder formulations intended for dry powder inhalation were successfully developed using TFF technology. A small amount of mannitol was used as a single excipient, and the aerosol performance of voriconazole nanoaggregates was advantageously enhanced by using phase-separated submicron crystalline mannitol as a surface texture modifier. Voriconazole dry powder for inhalation prepared by TFF can be used as a viable topical treatment option for invasive pulmonary aspergillosis, with high atomization efficiency and drug loading, while having potential benefits associated with the deposition of nanoaggregates in the airways.

[0120] Example 2 - Materials and General Methods

[0121] A. Materials

[0122] The following materials were purchased: voriconazole (Carbosynth, San Diego, CA); 25 (D-Basf, Ludwigshafen, Germany); acetonitrile (HPLC grade, Fisher Scientific, Pittsburgh, PA); trifluoroacetic acid (TFA) (HPLC grade, Fisher Scientific, Pittsburgh, PA); Tuffryn membrane filter (25 mm, 0.45 μm, Pall Corporation, Port Washington, NY). Filtered water (Evoqua, Warrendale, PA), pyrogen-free mannitol PF was donated by Roquette America Inc. (Geneva, IL).

[0123] B. Preparation of dry powder for inhalation using TFF

[0124] Mannitol and voriconazole (30%-100% w / w) powders were dissolved in a mixture of acetonitrile and water (50:50 v / v), and the solid content in the solution was maintained at 1% w / v. Approximately 15 μL of each solution was added dropwise to a rotating low-temperature cooling (-60°C) stainless steel drum from a height of 10 cm. The frozen samples were collected in a stainless steel container filled with liquid nitrogen and transferred to a -80°C freezer and then transferred to a freeze dryer. The solvent was removed using a VirTis Advantage freeze dryer (VirTis Company Inc., Gardiner, NY). The sample was placed at -40°C for 21 hours, the temperature was then slowly raised to 25°C over 21 hours, and placed at 25°C for another 21 hours to dry it. During the drying process, the pressure was maintained at 100 mTorr.

[0125] CX-Ray Powder Diffraction (XRPD)

[0126] The crystallinity of the powder samples was measured using an X-ray diffractometer (MiniFlex 600, Rigaku Co., Tokyo, Japan) in the angular range of 5-35° 2θ (step size 0.02°, 3° / min, 40 kV, 15 mA).

[0127] D. Scanning electron microscopy (SEM)

[0128] The surface morphology of the samples was identified using SEM (Zeiss Supra 40V SEM, Carl Zeiss, Heidenheim an der Brenz, Germany). An aliquot of the powder was placed on a carbon tape and sputter coated with 60 / 40 Pd / Au for 20 minutes before capturing the image.

[0129] E. Modulated Differential Scanning Calorimetry (mDSC)

[0130] Thermal analysis of powder samples was performed using a Q20 differential scanning calorimeter (TA Instruments, New Castle, DE) equipped with a cryogenic cooling system (RCS40, TA Instruments, New Castle, DE). In the modulated DSC, the modulation period was 50 s, the modulation amplitude was 1 °C, and the average heating rate was 5 °C / min. During the test, the samples were fixed using a Tzero pan and Tzero sealing cover manufactured by TA Instruments, and a hole was punched in the top cover with a 20G syringe needle before the pan was placed in the sample holder.

[0131] F. Scanning electron microscopy-energy dispersive X-ray spectroscopy (SEM / EDX)

[0132] The elements of the powders produced by TFF were identified using SEM / EDX (Hitachi S5500 SEM / STEM, Hitachi America, Tarrytown, NY).

[0133] G. Atomic force microscopy (AFM)

[0134] Two different types of atomic force microscopes were used during this study. Three-dimensional (3D) surface topography of particles produced by TFF was obtained using an Asylum MFP-3D AFM (Oxford Instruments, Oxfordshire, United Kingdom) equipped with an aluminum-coated MikroMasch HQ:NSC15 cantilever (NanoWorld AG, Switzerland) with a resonant frequency of 325kHz, a force constant of 40N / m, and a typical tip radius of 8nm. The powder was fixed to the AFM disc with carbon tape, and compressed nitrogen was used to blow off particles that were not firmly adhered to the carbon tape. The morphology analysis was performed in tapping mode with a scan rate of 1.00Hz, a set point of 1.08V, and an integral gain of 20.0. The feedback filter, drive amplitude, and drive frequency were optimized for each sample, and all images were collected at a resolution of 512×512. 3D morphology maps were generated using Gwyddion software (Necas and Klapetek, 2012) (64-bit Windows version 2.50).

[0135] To obtain images of nanoaggregates, a Park XE-100 AFM (Park systems, Suwon, Korea) equipped with an aluminum-coated Nanosensors PPP-NCHR cantilever (NanoWorld AG, Switzerland), with a resonant frequency of 330kHz, a force constant of 42N / m, and a tip radius of less than 7nm. First, a 380μm single-sided polished P-type silicon wafer was coated with 20 (VWR, Radnor, PA), and then loaded with powder samples for AFM analysis. 20 (1.5% w / v) was pre-dissolved in HPLC grade methanol (Fisher Scientific, Pittsburgh, PA). The solution was dripped onto a silicon wafer using a pipette and the solution was removed with compressed nitrogen. The powder was placed in a DP4 insufflator (Penn-Century Inc., Wyndmoor, PA) and atomized onto a silicon wafer using a 3 mL syringe. After the atomized powder was loaded onto the silicon wafer, compressed nitrogen was used to remove the powder solids that were not firmly adhered to the silicon wafer. Images were collected at a resolution of 512 × 512 and a scan rate of 0.30 Hz in tapping mode. Other values ​​of AFM were optimized for each sample. The morphology images were processed using Gwyddion software (Necas and Klapetek, 2012) (64-bit Windows version 2.50).

[0136] H. Aerodynamic Particle Size Distribution Analysis

[0137] Aerodynamic particle size was detected using a new generation pharmaceutical impactor (NGI) (MSP Co. Shoreview, MN), which was connected to a high capacity pump (HCP5 type, Copley Scientific, Nottingham, UK) and a critical flow controller (TPK2000 type, Copley Scientific, Nottingham, UK). #3 HPMC capsules (VCaps plus, Capsugel, Morristown, NJ) containing TFF powder (approximately 5-10 mg) were placed in a high resistance RS01 dry powder inhaler (Plastiape, Osnago, Italy) and dispersed into the NGI through the USP inhalation port, with a flow rate of 60 L / min, and each actuation lasting 4 seconds. In the entire test, no pre-separator was used. The NGI collection plate was coated with a methanol solution of 2% w / v polysorbate 20 and dried for 20 min before use. After atomization, the powder was extracted with a mixture of water and acetonitrile (50:50 v / v) and the content of voriconazole was analyzed by HPLC. Copley Inhaler Test Data Analysis Software (CITAS) version 3.10 (Copley Scientific, Nottingham, UK) was used to calculate the mass median aerodynamic diameter (MMAD), geometric standard deviation (GSD) and fine particle fraction (FPF) based on the deposition on the device, inhalation port, stage 1 to stage 7 and micropore collector (MOC).

[0138] I. High Performance Liquid Chromatography (HPLC)

[0139] Voriconazole content was measured using a Dionex Ultimate 3000 HPLC system (Sunnyvale, CA) and a Shimadzu DGU 14A degasser (Shimadzu, Kyoto, Japan). A Waters Xbridge C18 column (4.6 × 150 mm, 3.5 μm) (Milford, MA) was used. The details of the method are as follows: the aerodynamic properties were measured by an isocratic method with a mobile phase of 40 / 60 (v / v) water / acetonitrile (containing 0.1% (v / v) TFA) at a flow rate of 0.8 mL / min and an elution time of 4 min. In the stability study, a gradient method was used to analyze chemical degradation products. In the gradient method, acetonitrile solution containing 0.1% (v / v) TFA was gradually increased from 25% (v / v) to 95% (v / v) over 14 min and mixed with an aqueous solution containing 0.1% (v / v) TFA at a flow rate of 0.8 mL / min. In both methods, samples were analyzed at a detection wavelength of 254 nm and 25° C. Linearity was evaluated between 50 ng / mL and 100 Ug / mL using an injection volume of 15 μL.

[0140] J. Solution Nuclear Magnetic Resonance (Solution NMR)

[0141] use 1 The weight ratio between voriconazole and mannitol in TFF-VCZ-MAN powder was calculated by H NMR. 1 H NMR spectra were acquired at 25°C on a VNMR 600 (Varian, Palo Alto, CA) spectrometer at 600 MHz in the dimethyl sulfoxide- d6 (DMSO- d6 Chemical shifts were recorded relative to 2.47 ppm DMSO-d6.

[0142] K. Solid-state nuclear magnetic resonance (ssNMR)

[0143] ssNMR experiments were performed on a Bruker Avance III HD 400 MHz spectrometer (Bruker, Billerica, MA) at 25 °C with a magic angle spinning (MAS) frequency of 12 kHz. A Bruker 4 mm triple resonance HFX probe was used in dual resonance mode, tuned to 1 H / 13 C or 1 H / 19 F frequency. All samples were loaded into 4 mm ZrO2 rotors (Wilmad-LabGlass, PA) under ambient conditions. 13 C and 19In the cross-polarization (CP) MAS experiment, 1 The H channel was subjected to linearly increasing power levels in the range of 80-100 kHz and a contact time of 2 ms to enhance CP efficiency. High-power SPINAL64 proton decoupling was used at a field strength of 80 kHz. 13 C- 1 The same power parameters, contact time, and MAS frequency were used in the H CP heteronuclear correlation (HETCOR) experiment. Amantadine was used as the internal standard for calibration. 13 C chemical shift and referenced to the ethyl group at 38.48 ppm 13 Peak C.

[0144] L. Fourier transform infrared spectroscopy (FT-IR)

[0145] Using Smart OMNI-Sampler TM Nicolet TM iS TM The intermolecular activity between voriconazole and mannitol of TFF-VCZ-MAN powder was studied by 50 FT-IR (Thermo Fisher Scientific, Waltham, MA). The measurement was performed on dry powder samples and the spectrum was recorded in the range of 4000–700 cm at an aperture of 150, a resolution of 4, and 32 scans. -1 Spectrum inside.

[0146] M. Brunauer-Emmett-Teller (BET) specific surface area (SSA) analysis

[0147] Using Monosorb TM The SSA of TFF-VCZ-MAN powder was measured by single-point BET method using MS-21 rapid surface area analyzer (Quantachrome Instruments, Boynton Beach, FL). The samples were degassed in 20 psi nitrogen for 24 hours at ambient temperature to remove surface impurities. A mixture of nitrogen / helium (30:70 v / v) was used as the adsorption gas.

[0148] N. Shear resistance test

[0149] To test the shear resistance of TFF-VCZ-MAN 95:5 powder, the powder was placed in a stainless steel container (inner diameter 2 7 / 8 inches, height 4 1 / 4 inches) and pre-sheared by rolling the container at 85 rpm. Powder samples were collected at 15 min, 30 min and 60 min, and their aerodynamic properties were compared with the initial conditions.

[0150] O. Dissolution Test

[0151] The dissolution rate of voriconazole in the powder processed by TFF technology was quantitatively determined by in vitro dissolution method. The voriconazole released in the powder obtained by TFF was distinguished using Franz cell equipment. A new generation of pharmaceutical impactors (NGI) (MSP Co.Shoreview, MN) connected with a high-capacity pump (HCP5 type, Copley Scientific, Nottingham, UK) and a critical flow controller (TPK 2000 type, Copley Scientific, Nottingham, UK) was used to load atomized powder onto a Tuffryn membrane filter (25mm, 0.45 μm, Pall Corporation, Port Washington, NY). The five nozzles of the second stage on the NGI top cover were blocked with laboratory tape, leaving only one nozzle open. The Tuffryn membrane filter was placed and fixed on the collection cup below the nozzle opened at the second stage with laboratory tape. #3HPMC capsules (VCaps plus, Capsugel, Morristown, NJ) containing TFF powder (about 5-10 mg) were placed in a high resistance RS01 dry powder inhaler (Plastiape, Osnago, Italy) and dispersed into NGI through the USP inhalation port at a flow rate of 60 L / min, with each actuation lasting 4 seconds. No pre-separator was used. After atomization, the membrane filter loaded with powder (about 0.5-1 mg) was carefully removed from the collection cup and placed on top of the receiving chamber of the Franz cell, which had previously been filled with degassed 10 mM phosphate buffered saline (PBS, pH 7.4, 5 mL). The supply chamber was placed on the membrane filter, and the membrane filter was fixed between the receiving chamber and the supply chamber with a pinch clamp. The top of the supply chamber was covered with a sealing film. The dissolution test was carried out under a sink condition at 37°C, while constantly stirring with a magnetic stirring bar in the receiving chamber. The solvent (150 μL) was drawn at time intervals of 0 min, 20 min, 40 min, 60 min, 120 min and 180 min, and HPLC analysis was performed without dilution. After each sampling, fresh solvent was replaced.

[0152] P. Collection of atomized particles during dissolution and preparation of SEM samples

[0153] The SEM images of atomized TFF-VCZ-MAN powder before and after dissolution test were recorded using a rapid screening impactor (FSI) (Copley Scientific, Nottingham, UK) connected to a high capacity pump (HCP5 type, Copley Scientific, Nottingham, UK) and a critical flow controller (TPK 2000 type, Copley Scientific, Nottingham, UK). The #3HPMC capsule (VCaps plus, Capsugel, Morristown, NJ) comprising TFF powder (approximately 5-10mg) was put into a high resistance RS01 dry powder inhaler (Plastiape, Osnago, Italy), and was dispersed into a glass fiber filter (MSP Co.Shoreview, MN) arranged in the FSI to collect particles of 5 μm or less in aerodynamic diameter. After the particles were collected on the filter, the carbon tape on the filter was tapped, and these particles were transferred to the carbon tape attached to the SEM sample in advance, and the SEM images were recorded.

[0154] To record the SEM images during the dissolution test, the glass fiber filter loaded with powder on the FSI was cut into a circle (25 mm in diameter). Then at 37 ° C, the glass fiber filter was placed between the supply chamber and the receiving chamber of the Franz cell, which was pre-filled with PBS (pH 7.4). The filter was placed in the Franz cell for 5 min and then placed in a freezer at -80 ° C for 1 hour. At 25 ° C, the solvent was removed in 5 hours using a VirTis Advantage freeze dryer (VirTis Company Inc., Gardiner, NY). The carbon tape pre-attached to the SEM sample on the tapping glass fiber filter was used to transfer the TFF-VCZ-MAN powder, and the SEM images were recorded as described above.

[0155] Q. Stability study

[0156] TFF-VCZ-MAN 95:5 dry powder was pre-sheared in a glass bottle as described in the shear resistance test. 7.6 mg to 8.4 mg of pre-sheared powder was filled into #3HPMC capsules (Capsugel, Morristown, NJ). 14 capsules containing powder were transferred to scintillation vials, and the vials were purged with nitrogen for 20 s and then covered with lids. The vials were sealed in aluminum foil (13×15 cm), which was pre-purged with nitrogen for 30 s inside the vials and then stored at 25°C / 60% RH. Purity and aerosol performance tests were performed at various time points of 1, 3, 6, and 13 months.

[0157] R. Statistical analysis

[0158] Statistical analysis was performed using Student's t-test to compare aerodynamic performance and cumulative drug release rate. P values ​​< 0.05 were considered significant. 10.0.0 for significance of comparison data.

[0159] Example 3: Scale-up of voriconazole composition and inhaler testing

[0160] 1. Results

[0161] A. Monitor the cooling process at -60℃ and -150℃

[0162] Table 3 lists the different formulations and process conditions. Fig.16 Images of the freezing process at two different temperatures are shown. A water / ACN (50:50 v / v) solution containing voriconazole and mannitol (95:5 w / w) was used, wherein the solid loading was 1% (w / v) and 3% (w / v) (formulations #2, #4, #6 and #7 in Table 3). At -60°C, both solutions with different solid loadings showed that the freezing process was complete and reached thermal equilibrium in 200ms or less. Nucleation was observed at the edge of the sample disk at about 1 / 30ms, but freezing developed from the center of the disk to the edge at -60°C. In contrast, solutions with solid loadings of 1% (w / v) and 3% (w / v) began to nucleate at 1 / 60ms or less at -150°C and cooled uniformly throughout the sample disk. However, thermal equilibrium was not reached by 200ms.

[0163] Table 3: List of formulation and process parameters

[0164]

[0165] B. Physical properties of voriconazole nanoaggregates prepared by TFF

[0166] Figure 17 shows the high resolution morphology of voriconazole nanoaggregates treated at two different temperatures. It shows that the voriconazole nanoaggregates formed at a lower temperature (-150°C) (Formulation #4) are composed of smaller nanoparticles. When treated at -150°C, nanoparticles as small as 200nm were observed, while nanoparticles of about 500nm were found at -60°C (Formulation #2).

[0167] FIG18 compares the particle morphology of voriconazole nanoaggregates formed using different process parameters. When water / acetonitrile (30:70 v / v) (Formulation #1) was used as the solvent system, porous structures of mannitol with particle sizes exceeding 20 μm were observed. Voriconazole nanoaggregates prepared using other solvent systems showed surface texture modification of voriconazole nanoparticles by mannitol nanoparticles. Lower process temperatures resulted in smaller particles within the solid loading range tested (1% to 3% w / v).

[0168] FIG. 19 shows SEM images of aerosolized voriconazole nanoaggregates prepared at -60°C and -150°C (Formulations #7 and #6, respectively). This shows that the nanoaggregates consist of nanoparticles as small as 200 nm. Although the voriconazole nanoaggregates remain mainly micron-sized nanoaggregates, irregularly shaped nanoaggregates that are not completely deaggregated after aerosolization are observed. The surfaces of these nanoparticles still retain the modified texture after aerosolization through the DP4 insufflator.

[0169] C. Comparison of physical and aerodynamic properties with scaled-up products

[0170] Table 4 lists the aerodynamic properties and moisture content of voriconazole nanoaggregates prepared at small scale (200 mg) and large scale (90 g). Fig. 20 The crystallinity of various powder preparations was compared. The large-scale preparation product was compared with the small-scale preparation using There was no significant difference (p>0.05) in FPF (quantitative dose percentage, 35.6% and 37.0%, respectively), FPF (delivered dose percentage, 49.5% and 48.5%, respectively) and MMAD (3.69 μm and 3.52 μm, respectively) measured by the low resistance RS00 device at a flow rate of 60 L / min. In addition, TGA analysis results showed that the moisture content of both batches was less than 0.1% (w / w). The XRPD spectra of voriconazole nanoaggregates prepared on a small scale and on a large scale did not show any pattern differences.

[0171] Table 4. Comparison of physicochemical and aerodynamic properties by size

[0172]

[0173] D. In vitro aerosol performance

[0174] i. Co-solvents, process temperature and solid loading

[0175] The effects of co-solvents, process temperature and solid loading on the aerosol properties of unmodified voriconazole nanoaggregates were evaluated using NGI. The results are shown in Table 5. When the solid loading (1%) and process temperature (-60°C) were fixed (Formulations #1-3), different ratios of water to acetonitrile in the co-solvent changed the aerosol properties of voriconazole nanoaggregates. As the water ratio increased from 30% (v / v) to 50% (v / v) and 70% (v / v), FPF (quantitative dose percentage) increased from 34.3% to 37.9% and 45.6%. In addition, FPF (delivered dose percentage) increased (53.1%, 61.2% and 69.9%, respectively), while MMAD decreased (3.41 μm, 3.31 μm and 3.09 μm, respectively).

[0176] Table 5. Aerosol properties by solvent system, process temperature, and solids loading

[0177]

[0178] Plastiape low resistance RS-00 device, flow rate 60 L / min (n=3; mean ± SD) Unmodified powder

[0179] The effect of process temperature was also confirmed. When the process temperature was reduced from -60°C to -150°C, while the solid loading (1%) and co-solvent (water / ACN 50:50 v / v) were fixed (formulations #2 and #4), FPF (quantitative dose percentage) increased significantly from 37.9% to 46.7% (p<0.05). However, FPF (delivered dose percentage) and MMAD did not change significantly (61.2% and 67.5% and 3.31 μm and 3.27 μm, respectively) (p>0.05).

[0180] Solid loading also affects aerosol properties. Figure 5 As shown, higher solid loadings lead to lower aerosol properties. When the powder was unmodified, FPF (quantitative dose percentage) decreased from 46.7% to 41.3% and 37.0% as the solid loading increased from 1% to 2% and 3% (formulations #4-6). FPF (delivered dose percentage) also decreased from 67.5% to 60.9% and 48.5%. There was no significant difference in MMAD at solid loadings of 1% and 2% (formulations #4 and #5) (3.27μm and 3.24μm, respectively, p>0.05), while the MMAD at a solid loading of 3% (formulation #6) was significantly larger (3.52μm, p<0.05).

[0181] ii. By device

[0182] The aerosol performance of voriconazole nanoaggregates (formulation #6) was evaluated using four different types of Plastiape devices: low resistance and high resistance RS00 and low resistance and high resistance RS01. Table 6 shows the evaluation results of the effect of different concentrations on aerosol performance. The FPF (quantitative dose percentage) obtained at flow rates of 90L / min, 60L / min and 30L / min using the low resistance RS00 device was 48.6%, 45.8% and 27.0%, respectively, and the FPF (delivered dose percentage) was 63.7%, 63.9% and 48.9%, respectively. As the flow rate decreased from 90L / min to 60L / min and 30L / min, the MMAD increased from 3.22μm to 3.36μm and 4.32μm. The FPF (quantitative dose percentage) obtained at 60L / min and 30L / min using the high resistance RS00 device was 34.7% and 30.7%, respectively. The MMAD obtained at 60L / min and 30L / min using the high resistance RS00 device was 3.76μm and 3.83μm, respectively. The FPF (quantitative dose percentage) obtained at flow rates of 90L / min, 60L / min and 30L / min using the low resistance RS01 device was 40.1%, 35.8% and 27.0%, respectively, and the MMAD was 4.28μm, 4.37μm and 5.34μm, respectively. The FPF (quantitative dose percentage) obtained at flow rates of 60L / min and 30L / min using the high resistance RS01 was 31.7% and 20.2%, respectively, and the MMAD was 4.48μm and 5.06μm, respectively. In general, in in situ aerosol performance testing, higher aerodynamic performance is obtained at the same flow rate using a low resistance device, and the aerodynamic performance obtained using the RS00 device is better than the results obtained using RS01.

[0183] Table 6. Aerosol properties by device

[0184]

[0185]

[0186] iii. By dosage

[0187] The effects of different dose loads on aerosol performance were tested using high resistance RS00 and high resistance RS01 devices. The results are shown in Table 7. When the loading dose increased from 10 mg to 15 mg and 20 mg, the aerosol characteristics FPF (quantitative dose percentage) (34.7%, 33.8% and 31.8%, respectively), FPF (delivered dose percentage) (55.0%, 55.5% and 51.5%, respectively) and MMAD (3.76 μm, 3.77 μm and 3.84 μm, respectively) obtained at a flow rate of 60 L / min using the high resistance RS00 device did not change significantly (p>0.05). When the loading dose was 10 mg and 20 mg, there were significant differences (p<0.05) in FPF (quantified dose percentage) (31.7% and 25.3%, respectively), FPF (delivered dose percentage) (48.5% and 37.4%, respectively) and MMAD (4.48 μm and 5.21 μm, respectively) obtained using the high resistance RS01 device at a flow rate of 60 L / min.

[0188] Table 7. Aerosol properties by dose

[0189]

[0190] Table 8. Aerosols by Equipment Based on Pressure Drop

[0191]

[0192]

[0193] n = 3

[0194] Loading capacity: 15.0 mg / capsule (fixed)

[0195] 2. Discussion

[0196] A. Process design space for preparing voriconazole nanoaggregates using TFF

[0197] The process parameters within the design space of the freezing process used in TFF must be considered and their impact understood during development and subsequent scale-up, including: solvent system, process temperature, solid loading, and batch size. The process design space was determined using a low resistance RS00 device at a flow rate of 60 L / min, as nebulization using the low resistance RS00 device is more dependent on the inhalation flow rate and the characteristics of the formulation. This dependence can differentiate the nebulization of formulations made with different process design parameters.

[0198] B. Solvent System

[0199] The physicochemical properties of amorphous solid dispersions of danazol prepared using TFF were not affected by two different solvents (tert-butyl alcohol and acetonitrile) (Overhoff et al., 2007); however, the crystallinity, morphology, and aerosol performance of voriconazole with PVP K12 or PVP K30 produced using TFF were different, depending on the solvent composition (including water and 1,4-dioxane) (Beinbom et al., 2012). The co-solvent system of water and acetonitrile used in this study was used to develop tacrolimus and voriconazole formulations prepared using TFF (Watts et al., 2013; Moon et al., 2019).

[0200] Although no differences in crystallinity were observed, morphological differences were observed among the different solvent compositions. In addition, significant trends in aerodynamic properties were observed when the solvent composition was changed. Without powder adjustment, solvent compositions containing a higher proportion of water showed enhanced atomization. This result may be related to two factors: viscosity of the co-solvent system and low temperature phase separation.

[0201] In a co-solvent system of water and acetonitrile, viscosity increases with increasing water content (Thompson et al., 2006; Cunningham et al., 1967). During the freezing process, high viscosity may hinder the movement of molecules. Therefore, the molecules are more evenly distributed in the frozen state, and the solute concentration in the unfrozen channels may not increase significantly. The low viscosity of the solvent allows the molecules to move more during the freezing process, and molecular agglomeration may occur. As a result, the solute concentration in the unfrozen channels increases. Since the voriconazole powder prepared using TFFE is a crystalline nanoaggregate, the higher solute concentration may induce the formation of larger nanoparticles due to the shorter distance between molecules.

[0202] Although TFF involves ultrafast supercooling, the freezing process of the water / ACN solvent system still requires 200 ms at -60 °C. Therefore, with a freezing time of 200 ms, the use of low-viscosity solvents may lead to a higher degree of molecular aggregation, resulting in a decrease in aerosol performance. This trend was also observed by Beinborn et al. in a previous study on voriconazole prepared using TFF (Beinborn et al., 2012). When a binary solvent system of water and 1,4-dioxane was used to produce crystalline voriconazole powders containing PVP K12 or PVP K30, TFF particles prepared with 1,4-dioxane / water (20:80 v / v) achieved higher crystallinity than particles prepared with 1,4-dioxane / water (50:50 v / v). Although the viscosity of 1,4-dioxane is higher than that of water, the viscosity of 1,4-dioxane / water (20:80 v / v) is higher than that of 1,4-dioxane / water (50:50 v / v) (Besbes et al., 2009). Therefore, the viscosity of the co-solvent system is one of the factors affecting the performance of the aerosol after lyophilization.

[0203] Preventing low temperature phase separation is another possibility to enhance aerosol performance by using a co-solvent system with a higher water content. Co-solvent systems consist of water and acetonitrile, and when containing 35%-88% (v / v) acetonitrile, phase separation during freezing is well known (Gu et al., 1994; Zarzycki et al., 2006). After phase separation occurs below -1.32°C (Zarzycki et al., 2006), the unfrozen solvent will separate into an 88% (v / v) acetonitrile phase and a 65% (v / v) water phase, and the solute may move to the phase where the solute has a higher solubility (Gu et al., 1994).

[0204] This low-temperature phase separation occurred in Formulation #1, which was treated with water / ACN (30:70 v / v) at -60°C. 5% (w / w) mannitol in voriconazole nanoaggregates was used as a surface texture modifier (Moon et al., 2019). Therefore, the mannitol observed on the surface of crystalline voriconazole nanoaggregates was nanoparticles, as shown in the SEM images of other formulations in Figure 18. However, porous mannitol particles of approximately 20 μm were observed, with the same morphology as TFF-mannitol (Moon et al., 2019). When mannitol particles of this size were generated in Formulation #1, the effect of mannitol on surface texture modification was reduced because a smaller amount of mannitol was available for surface texture modification, resulting in poor atomization effect.

[0205] Although supercooling in TFF may minimize phase separation and produce very small ice channels due to ultra-fast freezing (Moon et al., 2016), the process temperature of -60°C can still achieve a small amount of phase separation of water / ACN (30:70 v / v), promote aggregation during freezing (up to 200 ms) and increase in concentration of mannitol. However, water / ACN (70:30 v / v) did not undergo phase separation during freezing (Gu et al., 1994; Zarzycki et al., 2006). Therefore, aggregation and concentration increase caused by low-temperature phase separation are unlikely to occur.

[0206] C. Process temperature

[0207] In addition to the solvent composition, the process temperature also affects the aerosol performance of crystalline voriconazole nanoaggregates made using TFF. Lower process temperatures lead to higher supercooling, which produces smaller ice channels and prevents particle growth (Overhoff et al., 2009; Engstrom et al., 2008). In this study, ultra-fast supercooling was used when the temperature was -150°C, which showed a much faster nucleation rate. This supercooling at -150°C prevented particle growth and produced smaller nanoparticles in the voriconazole nanoaggregates, which were observed by AFM and SEM to be composed of nanoparticles as small as 200nm. In contrast, when processed at -60°C, the particle size observed using AFM was around 500nm. When voriconazole nanoaggregates are composed of smaller nanoparticles, they are more likely to deaggregate into smaller particles during inhalation, thereby improving aerosol performance.

[0208] Interestingly, the enhancement in aerosol performance caused by higher supercooling at -150°C was comparable to the higher performance produced by the co-solvent system of water / ACN (70:30 v / v) at lower supercooling at -60°C. There was no significant difference (p>0.05) in FPF (percentage of quantitative dose) and MMAD between the two process conditions (formulations #3 and #4).

[0209] D. Solid Loading

[0210] Increasing solid loading is one way to reduce manufacturing time when producing powder formulations using TFF. However, higher solid loadings generally compromise aerosol performance. Without modification or physical shearing, the bulk density of voriconazole nanoaggregates prepared using a solid loading of 1% (w / v) was approximately 30 mg / cm 3 Therefore, to speed up the manufacturing process, the solid loading was increased to 3% (w / v), which corresponds to a bulk density of 30 mg / cm 3When the solid loading was increased to 3% (w / v), two optimized process parameters were initially applied: water / ACN (70 / 30 v / v) solvent system and a processing temperature of -150°C. However, due to the low solubility of voriconazole in water / ACN (70 / 30 v / v), a solid loading of 3% (w / v) was not applicable. Therefore, water / ACN (50 / 50 v / v) was selected.

[0211] Although the bulk density obtained with a solid loading of 3% (w / v) was similar to that obtained with 1% (w / v) after lyophilization, the result was a lower aerosol performance before powder conditioning. However, appropriate modification could improve the performance to be comparable to that of voriconazole nanoaggregates (Formulation #4) prepared with a solid loading of 1% (w / v) and optimized process parameters. Therefore, powder-conditioned Formulation #6 was used for aerosolization testing.

[0212] E. Batch size

[0213] Until recently, the TFF method still used a syringe or a separatory funnel to add the solution dropwise. As a result, the solution takes longer to freeze. This is a major obstacle to scaling up the TFF method. To speed up the freezing process, amorphous meloxicam was produced by the TFF method using a 2-channel peristaltic pump (Jermain et al., 2019). However, in this study for voriconazole, the number of channels was increased to 8 and the feed rate of the solution was optimized to 25 mL / min. At the same time, the rotation speed of the cryogenic drum was increased from 10 rpm to 20 rpm to avoid overlapping of the frozen sample trays at higher feed rates.

[0214] The increase in speed shortens the retention time of the frozen sample on the cryogenic drum. Before being collected in the tray containing liquid nitrogen, this time is shortened from 4s to 2s. However, due to the ultra-fast freezing of TFF, the freezing process usually requires less than a few hundred milliseconds (Overhoff et al., 2007), and it is expected that the increase in the speed of the cryogenic drum will not affect the freezing process. For formulation #6 in this study, nucleation occurs in less than 1 / 60s (see Figure 18), and the frozen sample reaches thermal equilibrium in less than 2s before being collected in the tray.

[0215] Ultrafast supercooling at -150°C accelerates the nucleation rate and increases the number of ice crystals formed (Rambhatla et al., 2004; Overhoff et al., 2009). As a result, uniform nucleation is observed throughout the frozen sample. In addition, the freezing process is independent of the preparation scale, and there are no significant differences in the results of the frozen samples, regardless of the scale, because the droplets are of similar size. With the overall similar freezing process, the physicochemical and aerodynamic properties of the products prepared at a small scale are comparable to those prepared at a larger scale.

[0216] After deciding to use a peristaltic pump in the TFF process scale-up, the capacity of the lyophilizer was also tested. The data in Table 4 also confirmed that there was no difference between lyophilizing 90 g of voriconazole nanoaggregates using a 3-layer AdVantage Pro lyophilizer and lyophilizing 200 mg of voriconazole nanoaggregates using a 1-layer AdVantage 2.0 lyophilizer. Therefore, when using a 3-layer AdVantage Pro lyophilizer, it is suitable to scale up the TFF process using a peristaltic pump at a feed solution flow rate of 25 mL / min.

[0217] F. Interaction between the device and voriconazole nanoaggregates using different devices and flow rates

[0218] In the development of a drug product delivered via a DPI, device design or selection is as important as formulation development in terms of aerosol performance. The same powder formulation can be aerosolized differently using different DPI devices (Parumasivam et al., 2017). This study tested the commercially available Plastiape RS01 and RS00 devices, which are applicable to many DPI products on the market or in development (Armer et al., 2016; Elkins et al., 2014; Roscigno et al., 2017). The RS01 and RS00 devices use the same delivery technology: the capsule is lifted from its housing and rotated at high speed (Dry Powder Inhaler RS01: How to Use: Plastiape; (available from: plastiape.com / en / content / 1635 / dry--powder-inhaler--rs01--how--use). However, the powder in the RS01 device empties the capsule through two holes, while the powder in the RS00 device is discharged through eight smaller holes of the capsule with a longer mouthpiece. In contrast, when compared using the same flow rate with the same type of resistance, the overall aerosol performance of voriconazole nanoaggregates using the RS00 device is superior.

[0219] This higher performance obtained using the RS00 device may be due to the smaller pores produced by the perforation system of the RS00 device. The smaller pores may help the larger voriconazole nanoaggregates to disaggregate when they leave the capsule, and their smaller size leads to smaller MMAD and higher FPF. This may be a unique feature of voriconazole nanoaggregates used in DPIs because they are composed of brittle nanoaggregates. Other powder formulations for DPIs made using spray drying or grinding may not be considered nanoaggregates. Therefore, the size of the pores may not have a significant impact on the overall performance when the powder exits the capsule.

[0220] Comparing the low-resistance and high-resistance RS01 and RS00 devices, the two low-resistance devices generally performed better than the high-resistance devices at flow rates of 60 L / min and 30 L / min. In addition, the low-resistance devices showed higher ED relative to the high-resistance devices. However, powder deagglomeration and microdispersion using low-resistance devices depend on the patient's inhalation flow rate (Dal Negro RW, 2015), resulting in changes in the nebulization effect at different inhalation flow rates. In this study, this result was also observed in the two low-resistance devices. Although the maximum degree of nebulization was achieved at a flow rate of 90 L / min using the low-resistance RS00 device, a significant decrease in FPF (percentage of quantitative dose) was observed at a flow rate of 30 L / min (18.8%). A similar trend was observed when using the low-resistance RS01 device. When using the low-resistance RS01, a significant decrease in FPF (percentage of quantitative dose) also occurred (8.8%) when the flow rate decreased from 60 L / min to 30 L / min. However, when using the high resistance RS00 device, the FPF (percentage of quantitative dose) differed by only 4.0% at flow rates of 60 L / min and 30 L / min, and although a significant difference in ED was observed, there was no significant difference in MMAD (p>0.05). The high resistance RS00 device exhibited inhalation flow rate independence between 60 L / min and 30 L / min caused by sufficient turbulence protocol (Dal Negro RW, 2015). Therefore, although the performance of the low resistance RS00 device is superior from the perspective of aerosol performance, these performances may vary greatly between individual patients, resulting in differences in efficiency. However, in the case of the high resistance RS01 device, flow rate independence between 60 L / min and 30 L / min was not observed, confirming that the smaller holes in the RS00 device facilitate the aerosolization of voriconazole nanoaggregates.

[0221] G. Different dosage loads

[0222] Prior to TFF freezing, the bulk density of voriconazole nanoaggregates before conditioning was typically about 30 mg / cm 3 , independent of the solid loading of the solution (1%-3% w / v). However, the bulk density gradually increases up to 100 mg / cm by conditioning or externally applied physical shear stress. 3 The voriconazole nanoaggregates were adjusted to have a density of about 60 mg / cm 3The bulk density of the voriconazole nanoaggregates was determined and the effect of powder loading was evaluated with the #3 HPMC capsule. Since the capacity of the #3 capsule is 0.3 mL, up to about 20 mg of the adjusted voriconazole nanoaggregates can be inserted into the capsule. Therefore, the aerosol performance of the voriconazole nanoaggregates was evaluated in the dose range of 10-20 mg per capsule. High resistance RS00 and RS01 devices were used at a flow rate of 60 L / min, and Tukey-Kramer HSD tests were performed to compare the results between different powder levels.

[0223] While the high resistance RS00 device showed no significant differences in FPF (percentage of dose delivered), FPF (percentage of dose delivered), and MMAD (p>0.05), the performance obtained using the high resistance RS01 device was different at doses of 10 mg and 20 mg (p<0.05). In the case of voriconazole nanoaggregates, the consistency of aerosol performance obtained using the high resistance RS00 device may be a result of the smaller pores facilitating the aerosolization of particles with a narrow size distribution.

[0224] 3. Materials and Methods

[0225] A. Materials

[0226] Voriconazole USP was purchased from Aurobino Pharma Ltd. (Hyderabad, India). HPLC grade acetonitrile (ACN), methanol, and trifluoroacetic acid (TFA) were purchased from Fisher Scientific (Pittsburgh, PA). In-house filtered water (Evoqua, Warrendale, PA), pyrogen-free mannitol, and PF was donated by Roquette America Inc. (Geneva, IL).

[0227] B. Preparation of Powder Formulations

[0228] Voriconazole (95% w / w) and mannitol (5% w / w) were dissolved in a mixture of acetonitrile and water (30:70, 50:50 or 70:30 v / v) to a solid content of 1%-3% (w / v) in the solution. The solution was sonicated until a clear solution was obtained. The solution was then dripped onto a rotating cryogenically cooled (-60°C or -150°C) stainless steel drum from a height of approximately 10 cm. In small-scale preparations, a 10 mL syringe with a syringe needle (18 gauge) was used to feed the solution onto the drum. In large-scale preparations, the solution was diluted with a 10 mL syringe with a syringe needle (18 gauge). High-performance precision platinum-cured silicone pump tubing (16 gauge, Cole-Parmer, Vernon Hills, IL) A peristaltic pump (Cole-Parmer, Vernon Hills, IL) delivers the solution to the drum at a flow rate of 25 mL / min. During the freezing process, the frozen samples were collected in a stainless steel freeze dryer tray equipped with liquid nitrogen and transferred to a -80 ° C freezer to remove excess liquid nitrogen, and then the samples were transferred to the freeze dryer. The solvent was sublimated using a VirTis Advantage 2.0 or VirTis Advantage Pro desktop tray freeze dryer (VirTis Company Inc., Gardiner, NY) and the samples were dried. During the primary drying process, the shelf was kept at -40 ° C for 20 h, and then the shelf temperature was linearly increased to 25 ° C within 20 h, and then kept at 25 ° C for 20 h. Secondary drying was performed at 25 ° C for 20 h. During the freeze drying process, the pressure was maintained at 100 mTorr.

[0229] CX-Ray Powder Diffraction (XRPD)

[0230] The powder crystallinity was identified using an X-ray diffractometer (MiniFlex 600, Rigaku Co., Tokyo, Japan) in the range of 5-40° 2θ (step size 0.02°, 2° / min, 40 kV, 15 mA).

[0231] D. Scanning electron microscopy (SEM)

[0232] The surface morphology of the powder samples was determined using SEM (Zeiss Supra 40V SEM, Carl Zeiss, Heidenheim an der Brenz, Germany), and the atomized nanoparticles were identified. To identify the surface morphology, an aliquot of the powder was placed on a carbon tape, and the image was captured after sputtering 20 nm with 60 / 40Pd / Au. To measure the nanoparticles, 1-2 mg of powder was placed in a DP4 insufflator (Penn-Century Inc., Wyndmoor, PA), and a 3 mL syringe was used to atomize it onto a 380 μm single-sided polished P-type silicon wafer, and the image was captured after sputtering 5 nm with 60 / 40Pd / Au.

[0233] E. Atomic force microscopy (AFM)

[0234] To obtain images of nanoaggregates, Asylum MFP-3D AFM (Oxford Instruments, Oxfordshire, United Kingdom) was used, which was equipped with a gold-plated MikroMasch Hi'Res-C15 / Cr-Au cantilever (nanoWorld AG, Neuchatel, Switzerland), with a resonant frequency of 325kHz, a force constant of 40N / m, and a typical tip radius of 1nm. The powder was fixed to a silicon wafer using a DP4 insufflator (Penn-Century Inc., Wyndmoor, PA). 1-2mg of powder was placed in the insufflator and atomized onto a 380μm single-sided polished P-type silicon wafer using a 3mL syringe. After loading the powder, compressed nitrogen was used to blow off the excess powder that was not firmly adhered to the wafer. A scan rate of 1.00Hz was used for morphology analysis in tapping mode. Other values ​​of AFM were optimized for each sample. Images were acquired using 512 x 512 resolution and processed using Gwyddion software (version 2.50 for 64-bit Windows) (Necas and Klapetek, 2012).

[0235] F. Powder adjustment

[0236] 1.3 g of powder was added to a 60 mL Pyrex bottle. The sample bottle was rotated at 60 rpm for 30 min to shear the powder and stored in desiccant at room temperature.

[0237] G. Aerodynamic Particle Size Distribution Analysis

[0238] The aerodynamic properties of the powders were measured using a new generation pharmaceutical impactor (NGI) (MSP Co. Shoreview, MN) equipped with a critical flow controller (TPK model, MSP Corporation, Shoreview, MN) and a high volume pump (HCP5 model, MSP Corporation, Shoreview, MN). Approximately 5-20 mg of the powder formulation was inserted into a #3 HPMC capsule ( plus, The voriconazole content was analyzed by HPLC. The voriconazole content was analyzed by HPLC. Copley Inhaler Test Data Analysis Software (CITAS) 3.10 (Copley Scientific, Nottingham, UK) was used to calculate MMAD, geometric standard deviation (GSD) and fine particle fraction (FPF).

[0239] H. High performance liquid chromatography (HPLC)

[0240] For quantitative analysis of voriconazole content, a Dionex Ultimate 3000 HPLC system (Sunnyvale, CA) connected to a Shimadzu DGU 14A degasser (Shimadzu, Kyoto, Japan) and a Waters Xbridge C18 column (4.6×150 mm, 3.5 μm) (Milford, MA) was used. An isocratic method was used with a mobile phase of 40 / 60 (v / v) water / acetonitrile containing 0.1% (v / v) TFA at a flow rate of 0.8 mL / min for 4 min at 25°C. Sample concentrations were determined using a wavelength of 254 nm. A linearity study was performed for the standard curve between 62.5 ng / mL and 500 μg / mL of voriconazole using an injection volume of 7 μL.

[0241] I. Brunauer-Emmett-Teller (BET) Specific Surface Area (SSA) Analysis

[0242] To measure SSA, Monosorb TM MS-21 rapid surface area analyzer (Quantachrome Instruments, Boynton Beach, FL). Powder formulation samples were degassed using 20 psi nitrogen at ambient temperature for more than 24 h. A mixture of nitrogen and helium (30:70 v / v) was used as the adsorption gas.

[0243] J. Thermogravimetric analysis (TGA)

[0244] TGA was performed to measure the moisture content in the powder formulations. A Mettler TGA / DSC 1 thermogravimetric analyzer (Columbus, OH) was used. About 2-5 mg of the sample was placed in an alumina crucible (Mettler-Toledo, Columbus, OH) and the crucible lid was closed. The crucible was heated from 25°C to 150°C at a heating rate of 5°C / min. The moisture content of the sample was calculated by comparing the reduction in sample weight between 25°C and 125°C.

[0245] K. Photos frozen at different temperatures

[0246] To monitor the difference in freezing rate between different process temperatures in TFF, images of the freezing process were captured using a Canon EOS Rebel SL1 DSLR camera (Canon USA, Melville, NY) equipped with an 18-55 mm IS STM lens (Canon USA, Melville, NY) at a frame rate of 60 frames / sec and a resolution of 1280 × 720. The captured images were cropped to approximately 200 × 200 to show only the sample.

[0247] L. Statistical analysis

[0248] The Student's t test was used to determine whether there were statistical differences in aerodynamic properties. P values ​​< 0.05 were considered significant. 10.0.0 Calculate the p-value of the data.

[0249] ***

[0250] In view of the present disclosure, all compositions and methods disclosed and claimed herein can be made and performed without undue experimentation. Although the compositions and methods of the present disclosure have been described according to preferred embodiments, it will be apparent to those skilled in the art that changes may be applied to the methods described herein and the steps of the methods or the order of the steps without departing from the concept, spirit and scope of the present disclosure. More specifically, it will be apparent that certain agents that are both chemically and physiologically related can replace the agents described herein while achieving the same or similar results. All such similar substitutions and modifications apparent to those skilled in the art are considered to fall within the spirit, scope and concept of the present disclosure as defined by the appended claims.

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Claims

1. A pharmaceutical composition comprising: (A) a therapeutic agent; wherein the therapeutic agent is an antifungal agent, and wherein the therapeutic agent is in a crystalline state; and (B) an excipient, wherein the excipient accounts for 3% w / w to 10% w / w of the pharmaceutical composition; and wherein the excipient is a sugar alcohol or an amino acid; wherein the pharmaceutical composition is formulated as a dry powder as nanoaggregates, the nanoaggregates comprising nanoparticles of the therapeutic agent, and the surfaces of the nanoparticles of the therapeutic agent comprising discrete domains of the excipient, and wherein the discrete domains of the excipient reduce the contact area between the nanoparticles of the therapeutic agent.

2. The pharmaceutical composition according to claim 1, wherein the excipient comprises 3% w / w to 9% w / w of the pharmaceutical composition.

3. The pharmaceutical composition according to claim 2, wherein the excipient comprises 3% w / w to 6% w / w of the pharmaceutical composition.

4. The pharmaceutical composition according to claim 3, wherein the excipient comprises 3% w / w of the pharmaceutical composition.

5. The pharmaceutical composition according to claim 3, wherein the excipient comprises 5% w / w of the pharmaceutical composition.

6. A pharmaceutical composition according to any one of claims 1-5, wherein the discrete domains of excipient comprise one or more discontinuous domains of the excipient on the surface.

7. A pharmaceutical composition according to any one of claims 1-5, wherein the discrete domains of excipient comprise adjacent and continuous excipient layers.

8. The pharmaceutical composition according to claim 1, wherein the excipient is a sugar alcohol.

9. The pharmaceutical composition according to claim 8, wherein the excipient is mannitol.

10. The pharmaceutical composition according to claim 1, wherein the excipient is an amino acid. The pharmaceutical composition according to claim 10 , wherein the amino acid is leucine.

12. The pharmaceutical composition of claim 1, wherein the excipient is present as a nanodomain in the pharmaceutical composition.

13. The pharmaceutical composition of claim 12, wherein the nano-domains of the excipient have a size of 50 nm to 500 nm.

14. The pharmaceutical composition according to claim 13, wherein the size of the nano-domains of the excipient is 100 nm to 200 nm.

15. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition has a mass median aerodynamic diameter of 1.5 μm to 7.5 μm.

16. The pharmaceutical composition according to claim 15, wherein the mass median aerodynamic diameter is 2.5 μm to 6.5 μm.

17. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition does not contain a wax excipient.

18. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition does not comprise a hydrophobic excipient.

19. The pharmaceutical composition according to claim 1, wherein the antifungal agent is an azole antifungal drug.

20. The pharmaceutical composition according to claim 19, wherein the azole antifungal drug is voriconazole.

21. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises one or more additional excipients.

22. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition further comprises one or more additional therapeutic agents.

23. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is formulated for topical administration.

24. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is formulated for parenteral administration.

25. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is formulated for oral, intraadipose, intraarterial, intraarticular, intracranial, intramuscular, intranasal, intraocular, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrathecal, intratracheal, intratumoral, intraumbilical, intravenous, intracapsular, mucosal, rectal, subconjunctival, subcutaneous, sublingual, topical, vaginal; in the form of an emulsion, in the form of a lipid composition; via infusion or via inhalation.

26. The pharmaceutical composition of claim 25, wherein the pharmaceutical composition is formulated for administration via inhalation.

27. The pharmaceutical composition of claim 1, wherein the pharmaceutical composition is formulated for use with an inhaler.

28. The pharmaceutical composition according to claim 27, wherein the inhaler is a fixed dose combination inhaler.

29. The pharmaceutical composition of claim 28, wherein the inhaler is a metered dose inhaler.

30. The pharmaceutical composition according to claim 28 or 29, wherein the inhaler is a capsule-type inhaler.

31. The pharmaceutical composition of claim 27, wherein the inhaler is a low resistance inhaler.

32. The pharmaceutical composition of claim 27, wherein the inhaler is a high resistance inhaler.

33. The pharmaceutical composition of claim 27, wherein the inhaler is used at a flow rate of 10 L / min to 150 L / min.

34. The pharmaceutical composition of claim 33, wherein the flow rate is 20 L / min to 100 L / min.

35. The pharmaceutical composition of claim 27, wherein the inhaler has a pressure differential of 0.5 kPa to 5 kPa.

36. The pharmaceutical composition of claim 35, wherein the pressure differential is 1 kPa, 2 kPa or 4 kPa.

37. The pharmaceutical composition of claim 27, wherein the inhaler has a loading dose of 0.1 mg to 50 mg.

38. The pharmaceutical composition of claim 37, wherein the inhaler has a loading dose of 0.1 mg to 10 mg.

39. The pharmaceutical composition of claim 37, wherein the inhaler has a loading dose of 5 mg to 50 mg.

40. The pharmaceutical composition of claim 39, wherein the loading dose is 5 mg to 25 mg.

41. The pharmaceutical composition of claim 27, wherein the inhaler is configured to deliver one or a series of doses from one or more unit doses loaded sequentially.

42. The pharmaceutical composition of claim 41, wherein the inhaler is configured to deliver one dose from one unit dose.

43. The pharmaceutical composition of claim 41, wherein the inhaler is configured to deliver a series of doses from one unit dose.

44. The pharmaceutical composition of claim 41, wherein the inhaler is configured to deliver a dose each from a series of capsules loaded sequentially.

45. The pharmaceutical composition of claim 41, wherein the inhaler is configured to deliver a series of doses from a series of capsules loaded sequentially.

46. ​​Use of a pharmaceutical composition according to any one of claims 1 to 45 in the preparation of a medicament for preventing or treating a fungal infection in a patient in need thereof.

47. The use according to claim 46, wherein the patient's disease or disorder is in the lungs.

48. The use according to claim 46, wherein the therapeutic agent is an azole antifungal agent.

49. The use according to claim 48, wherein the therapeutic agent is voriconazole.

50. A method for preparing a pharmaceutical composition according to any one of claims 1-45, comprising: (A) mixing a therapeutic agent and an excipient with a solvent to form a precursor solution, wherein the therapeutic agent is an antifungal agent and the excipient is a sugar alcohol or an amino acid; (B) depositing the precursor solution onto a surface, the surface being at a temperature suitable to cause freezing of the solvent; and (C) removing the solvent to obtain a dry powder pharmaceutical composition.

51. The method of claim 50, wherein the solvent is a mixture of two or more solvents.

52. The method of claim 51, wherein the mixture of solvents comprises water.

53. The method of claim 52, wherein the solvent is an organic solvent.

54. The method of claim 53, wherein the organic solvent is acetonitrile.

55. The method of claim 53, wherein the organic solvent is 1,4-dioxane.

56. The method of claim 50, wherein the solvent is a mixture of water and an organic solvent.

57. The method of claim 56, wherein the solvent is a mixture of water and acetonitrile.

58. The method of claim 51, wherein the mixture of two or more solvents comprises 10% v / v to 90% v / v of organic solvent.

59. The method of claim 58, wherein the mixture comprises 40% v / v to 60% v / v of the organic solvent.

60. The method of claim 59, wherein the mixture comprises 50% v / v of the organic solvent.

61. The method of claim 58, wherein the mixture comprises 20% v / v to 40% v / v of the organic solvent.

62. The method of claim 61, wherein the mixture comprises 30% v / v of the organic solvent.

63. The method of claim 50, wherein the therapeutic agent and excipient comprise less than 10% w / v of the precursor solution.

64. The method of claim 63, wherein the therapeutic agent and excipient comprise 0.5% w / v to 5% w / v of the precursor solution.

65. The method of claim 64, wherein the therapeutic agent and excipient comprise 1% w / v of the precursor solution.

66. The method of claim 64, wherein the therapeutic agent and excipient comprise 3% w / v of the precursor solution.

67. The method of claim 50, wherein the surface is rotated.

68. The method of claim 50, wherein the temperature is from 0°C to -200°C.

69. The method of claim 68, wherein the temperature is from 0°C to -120°C.

70. The method of claim 69, wherein the temperature is between -50°C and -90°C.

71. The method of claim 70, wherein the temperature is -60°C.

72. The method of claim 69, wherein the temperature is -125°C to -175°C.

73. The method of claim 70, wherein the temperature is -150°C.

74. The method of claim 50, wherein the solvent is removed under reduced pressure.

75. The method of claim 74, wherein the solvent is removed via lyophilization.

76. The method of claim 75, wherein the lyophilization is performed at a lyophilization temperature of -20°C to -100°C.

77. The method of claim 76, wherein the lyophilization temperature is -40°C.

78. The method of claim 74, wherein the reduced pressure is less than 250 mTorr.

79. The method of claim 78, wherein the reduced pressure is 100 mTorr.

80. The method of claim 50, wherein the method further comprises heating the pharmaceutical composition under reduced pressure.

81. The method of claim 80, wherein the pharmaceutical composition is heated to a temperature of 0°C to 30°C.

82. The method of claim 81, wherein the temperature is room temperature or 25°C.

83. The method of claim 80, wherein the reduced pressure is less than 250 mTorr.

84. The method of claim 83, wherein the reduced pressure is 100 mTorr.

85. The method of claim 80, wherein the reduced pressure is the same as the reduced pressure during lyophilization.

86. A pharmaceutical composition comprising: (A) voriconazole, wherein the voriconazole is in a crystalline state; and (B) an excipient, wherein the excipient is mannitol, leucine or magnesium stearate, wherein the excipient accounts for 3% w / w to 10% w / w of the pharmaceutical composition; wherein the pharmaceutical composition is formulated as a dry powder as nanoaggregates, the nanoaggregates comprising nanoparticles of the voriconazole, and the surfaces of the nanoparticles of the voriconazole comprise discrete domains of the excipient, and wherein the discrete domains of the excipient reduce the contact area between the nanoparticles of the voriconazole.

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