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

Abstract

To provide pharmaceutical compositions which exhibit improved properties such as enhanced aerosolization ability and aerodynamic performance.SOLUTION: A pharmaceutical composition comprises: (A) a therapeutic agent; and (B) an excipient, where the excipient accounts for less than 10 wt.% of the pharmaceutical composition, and the therapeutic agent accounts for at least 90 wt.% of the pharmaceutical composition. The pharmaceutical composition is formulated as a dry powder as a nanoaggregate comprising nanoparticles of the therapeutic agent. The surface of the nanoparticles of the therapeutic agent comprises discrete domains of the excipient. The discrete domains of the excipient reduce the contact area between the nanoparticles of the therapeutic agent.SELECTED DRAWING: Figure 3-1

Description

[Technical field]

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

[0002] 1. Field The present disclosure relates generally to the field of pharmaceuticals and pharmaceutical manufacturing. More specifically, the present disclosure relates to compositions and methods for preparing drug compositions that include small amounts of excipients and therapeutic agents formulated as nanoaggregates. [Background technology]

[0003] 2. Description of Related Technology Until recently, delivery of aerosolized antifungals to the lungs was limited to amphotericin B (Le and Schiller, 2010; Borro et al. 2008). However, Hilberg et al. 2008 reported that inhaled voriconazole was more effective than inhaled amphotericin B for the treatment of invasive pulmonary aspergillosis (IPA), thereby supporting the successful treatment of patients with IPA who had previously failed oral or injectable forms of voriconazole, with or without inhaled amphotericin B.

[0004] Tolman et al. reported inhaled voriconazole delivered to the lungs by nebulization (Tolman et al. 2009a; Tolman et al. 2009b). However, the concentration of voriconazole in the lung tissue decreased to levels below the minimum detectable range 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 using sodium sulfobutylether-β-cyclodextrin (SBECD) as an excipient. The safety of SBECD delivered by the pulmonary route has not yet been confirmed, and this large amount of inactive ingredients can cause serious side effects (Wong 1993). Voriconazole formulations for dry powder inhalation (DPI) have been reported by Sinha et al. using polylactide-co-glycolide nanoparticles (Sinha et al. 2013) and Arora et al. using polylactide microparticles (Arora et al. 2015), but the drug loading of these particles was low (31% and 20% w / w, respectively). Arora et al. reported another voriconazole powder formulation for DPI using leucine as an excipient (Arora et al. 2016). However, all of these DPI powder formulations contain non-GRAS excipients not previously used in FDA-approved inhalation drugs. Beinborn et al. also used thin film freezing (TFF), a particle engineering technique, to develop amorphous and crystalline voriconazole formulations suitable for dry powder inhalation (Beinborn et al. 2012a; Beinborn et al. 2012b). However, the amorphous formulation contained 75% (w / w) excipients, which resulted in low efficacy, poor drug absorption, and rapid clearance based on in vivo pharmacokinetic data in a mouse model. 0-24h is the AUC of the amorphous formulation 0-24h The FPFs of the crystalloid formulations were significantly higher in both lung (452.6 μg h / g and 232.1 μg h / g, respectively) and plasma (38.4 μg h / g and 18.6 μg h / g, respectively) than in the control. However, the aerosol performance of the crystalloid formulation was poorer (FPF 37.8%).

[0005] Recently, it was proposed based on modeling that nanoaggregates containing drug nanoparticles would be favorably distributed with higher lung epithelial coverage compared to discrete micron-sized particles and nanoparticles (Longest and Hindle 2017). Aggregates are particulate solid materials composed of collections of particles that are held together by strong inter- or intra-molecular cohesive forces (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 nanoaggregates of fluticasone propionate was 57 times higher than that of conventional microparticles. Although true nanoaerosols achieved better absorption efficiency, due to the lack of practical devices available to deliver true nanoaerosols to the peripheral airways, nanoaggregates offered the best approach to target the drug to the peripheral airways. Slowly dissolving nanoaggregates were described to show improved drug uptake and distribution based on Longest et al. (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). With high degrees of supercooling, TFF has been successfully used to produce nanostructured aggregates (Sinswat et al. 2008). Spray drying is another common technique to produce micro- or nano-scale particles for DPIs. However, particle formation during the drying process of spray drying is generally slower than the freezing process of TFF (Wisniewski 2015), resulting in longer particle growth times and larger particles. Thus, typical spray drying methods do not offer the benefits of increased uptake and microdosimetry that nanoaggregates have, as described by Longest and Hindle 2017. Thus, there remains a need to develop additional pharmaceutical compositions as nanoaggregates that exhibit improved properties, such as enhanced aerosolization. Summary of the Invention

[0007] The present disclosure provides pharmaceutical compositions as nanoaggregates comprising a therapeutic agent and an excipient, methods for their manufacture, and methods for their use. In some embodiments, the present disclosure provides a pharmaceutical composition comprising: (A) a therapeutic drug; and (B) an excipient, comprising less than about 10% by weight of the pharmaceutical composition; Including, Here, a pharmaceutical composition is provided in which the pharmaceutical composition is formulated as a nanoaggregate comprising nanoparticles of a therapeutic agent, the surface of the nanoparticles of the therapeutic agent comprising distinct domains of an excipient, the distinct domains of the excipient reducing the contact area between the nanoparticles of the therapeutic agent.

[0008] In some embodiments, the therapeutic agent is present in a crystalline form. In other embodiments, the therapeutic agent is present in an amorphous form. In some embodiments, the excipient comprises about 9% w / w to about 1% w / w of the pharmaceutical composition, e.g., 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.

[0009] In some embodiments, the separate domains of the excipient comprise one or more discontinuous domains of the excipient on the surface. In other embodiments, the separate domains of the excipient comprise adjacent continuous layers of the excipient. In some embodiments, the excipient is water soluble. In some embodiments, the excipient is solid at room temperature. In some embodiments, the excipient is a sugar alcohol, e.g., mannitol. In some embodiments, the excipient is present as nanodomains in the pharmaceutical composition. In some embodiments, the nanodomains of the excipient have a size of about 50 nm to about 500 nm, e.g., about 100 nm to about 200 nm.

[0010] In some embodiments, the pharmaceutical composition has a mass median aerodynamic diameter of about 1.5 to about 7.5 μm, e.g., about 2.5 to about 6.5 μm. In some embodiments, the pharmaceutical composition does not include a wax excipient. In some embodiments, the pharmaceutical composition does not include a hydrophobic excipient. In some embodiments, the therapeutic agent is an anticancer agent, an antifungal agent, a psychotropic agent, e.g., an analgesic agent, an agent that alters the level of consciousness, e.g., an anesthetic or hypnotic agent, a nonsteroidal anti-inflammatory drug (NSAIDS), an anthelmintic, a beta agonist, an antiacne agent, an antiangina agent, an antiarrhythmic agent, an antiasthma agent, an antibacterial agent, an antibenign prostatic hyperplasia agent, an anticoagulant, an antidepressant, an antidiabetic agent, an antiemetic agent, an antiepileptic agent, an antigout agent, an antihypertensive agent, an antiinflammatory agent, an antimalarial agent, an antimigraine agent, an antimuscarinic agent, an antitumor agent, an antiobesity agent, an antiosteoporosis agent, an antiperfusion agent, an anti- ...-perfusion agent, an anti-inflammatory agent, an anti-inflammatory agent, an anti-malarial agent, an antimigraine agent, an antimuscarinic agent, an antitumor agent, an antiobesity agent, an antiosteoporosis agent, an anti-perfusion agent, an anti-perfusion agent, an anti-per The therapeutic agent is selected from the group including: kinson's disease drugs, antiproliferative drugs, antiprotozoal drugs, antithyroid drugs, antitussive drugs, antiurinary incontinence drugs, antiviral drugs, anxiolytic drugs, appetite suppressants, beta blockers, cardiac inotropic drugs, chemotherapy drugs, cognitive enhancers, contraceptive drugs, corticosteroids, Cox-2 inhibitors, diuretics, erectile dysfunction improving drugs, expectorants, gastrointestinal drugs, histamine receptor antagonists, immunosuppressants, keratolytic drugs, lipid regulating drugs, leukotriene inhibitors, macrolides, muscle relaxants, neuroleptics, nutritional agents, opioid analgesics, protease inhibitors, and sedatives. In some embodiments, the therapeutic agent is an antifungal agent, for example, 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.

[0011] In some embodiments, the pharmaceutical composition is formulated for oral administration, intraadipose administration, intra-arterial administration, intra-articular administration, intracranial administration, intradermal administration, intralesional administration, intramuscular administration, intranasal administration, intraocular administration, intrapericardial administration, intraperitoneal administration, intrapleural administration, intraprostatic administration, intrarectal administration, intrathecal administration, intratracheal administration, intratumoral administration, intraumbilical administration, intravaginal administration, intravenous administration, intravesical administration, intravitreal administration, liposomal administration, local administration, mucosal administration, parenteral administration, rectal administration, subconjunctival administration, subcutaneous administration, sublingual administration, topical administration, transdermal administration, vaginal administration, in the form of a cream, in the form of a lipid composition, administration by catheter administration, administration by irrigation administration, administration by continuous infusion administration, administration by injection administration, administration by inhalation administration, administration by injection administration, administration by local delivery administration, or administration by local perfusion administration. In some embodiments, the pharmaceutical composition is formulated for administration by inhalation administration.

[0012] In some embodiments, the pharmaceutical composition is formulated for use in an inhaler, such as a fixed dose combination inhaler, a single dose dry powder inhaler, a multi-dose dry powder inhaler, a multi-unit dose dry powder inhaler, a metered dose inhaler, or a pressurized metered dose inhaler. In some embodiments, the inhaler is a capsule-based inhaler. In some embodiments, the inhaler is a low resistance inhaler. In other embodiments, the inhaler is a high resistance inhaler. In some embodiments, the inhaler is used at a flow rate of about 10 L / min to about 150 L / min, such as about 20 L / min to about 100 L / min. In some embodiments, the inhaler has a pressure differential of 0.5 kPa to about 5 kPa. In some embodiments, the pressure differential is 1 kPa, 2 kPa, or 4 kPa. In some embodiments, the inhaler has a loading dose of about 0.1 mg to about 50 mg. In some embodiments, the inhaler has a loading dose of about 0.1 mg to about 10 mg. In other embodiments, the inhaler has a loading dose of about 5 mg to about 50 mg, for example about 5 mg to about 25 mg. In some embodiments, the inhaler is configured to deliver one or a series of doses from one or more unit doses loaded in series. In some embodiments, the inhaler is configured to deliver one dose from one unit dose. In other embodiments, the inhaler is configured to deliver a series of doses from one unit dose. In other embodiments, the inhaler is configured to deliver one dose each from a series of capsules loaded in series. In other embodiments, the inhaler is configured to deliver a series of doses from a series of capsules loaded in series.

[0013] In yet another aspect, the disclosure provides a method of treating or preventing a disease or disorder in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition described herein that includes a therapeutic agent effective to treat the disease or disorder. In some embodiments, the disease or disorder is in the lungs. In some embodiments, the disease or disorder is an infection, e.g., a fungal infection. In some embodiments, the therapeutic agent is an antifungal agent, e.g., an azole antifungal agent. In some embodiments, the therapeutic agent is voriconazole.

[0014] In yet another aspect, the present disclosure provides a method for preparing a pharmaceutical composition, comprising: (A) mixing a therapeutic agent and excipients present in an amount less than 10% w / w with a solvent to form a precursor solution; (B) depositing the precursor solution onto the surface at a temperature suitable to freeze the solvent; and (C) Removing the solvent to obtain a pharmaceutical composition The present invention provides a method comprising:

[0015] In some embodiments, the solvent is a mixture of two or more solvents. In some embodiments, the mixture of solvents includes 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, the mixture of two or more solvents includes about 10% v / v to about 90% v / v of the organic solvent. In some embodiments, the mixture includes about 40% v / v to about 60% v / v of the organic solvent, such as about 50% v / v of the organic solvent. In other embodiments, the mixture includes about 20% v / v to about 40% v / v of the organic solvent, such as about 30% v / v of the organic solvent. In some embodiments, the therapeutic agent and excipients comprise less than 10% w / v of the precursor solution, such as about 0.5% to about 5% w / v of the precursor solution. In some embodiments, the therapeutic agent and excipients comprise about 1% w / v of the precursor solution. In other embodiments, the therapeutic agent and excipients comprise about 3% w / v of the precursor solution.

[0016] 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., e.g., 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., e.g., about −150° C. In some embodiments, the solvent is removed under reduced pressure. In some embodiments, the solvent is removed by lyophilization. In some embodiments, the lyophilization is performed at a lyophilization temperature of about −20° C. to about −100° C., e.g., about −40° C. In some embodiments, the reduced pressure is less than 250 mTorr, e.g., about 100 mTorr.

[0017] In some embodiments, the method further comprises 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., e.g., near room temperature or about 25° C. In some embodiments, the reduced pressure is less than 250 mTorr, e.g., about 100 mTorr. In some embodiments, the reduced pressure is the same as the reduced pressure during lyophilization.

[0018] In yet another aspect, the disclosure provides pharmaceutical compositions prepared according to the methods described herein.

[0019] [The present invention 1001] A pharmaceutical composition, comprising: (A) a therapeutic drug; and (B) an excipient, comprising less than about 10% by weight of the pharmaceutical composition; Including, wherein the pharmaceutical composition is formulated as nanoaggregates comprising nanoparticles of the therapeutic agent, the surfaces of the nanoparticles of the therapeutic agent comprising distinct domains of the excipient, the distinct domains of the excipient reducing the contact area between the nanoparticles of the therapeutic agent. The pharmaceutical composition. [The present invention 1002] The pharmaceutical composition of any one of claims 1 to 10, wherein said therapeutic agent is present in a crystalline form. [The present invention 1003] The pharmaceutical composition of any one of claims 1 to 10, wherein said therapeutic agent is present in amorphous form. [The present invention 1004] The pharmaceutical composition according to any one of claims 1001 to 1003, wherein said excipient constitutes about 9% w / w to about 1% w / w of said pharmaceutical composition. [The present invention 1005] 1004. The pharmaceutical composition of the present invention, wherein said excipient constitutes about 6% w / w to about 2% w / w of said pharmaceutical composition. [The present invention 1006] 1005. The pharmaceutical composition of claim 10, wherein said excipient comprises about 3% w / w of said pharmaceutical composition. [The present invention 1007] The pharmaceutical composition of the present invention, wherein said excipient comprises about 5% w / w of said pharmaceutical composition. [The present invention 1008] The pharmaceutical composition of any of claims 1001 to 1007, wherein said distinct domains of said excipient comprise one or more discontinuous domains of said excipient on said surface. [The present invention 1009] The pharmaceutical composition of any one of claims 1001 to 1007, wherein said distinct domains of said excipient comprise adjacent contiguous layers of said excipient. [The present invention 1010] The pharmaceutical composition of any one of claims 1001 to 1009, wherein the excipient is water-soluble. [The present invention 1011] The pharmaceutical composition of any one of claims 1001 to 1010, wherein the excipient is a sugar alcohol. [The present invention 1012] The pharmaceutical composition of the present invention, wherein said excipient is mannitol. [The present invention 1013] The pharmaceutical composition according to any one of claims 1001 to 1012, wherein the excipient is present in the pharmaceutical composition as a nano-domain. [The present invention 1014] The pharmaceutical composition of the present invention, wherein the nano-domains of the excipient have a size of about 50 nm to about 500 nm. [The present invention 1015] 1014. The pharmaceutical composition of the present invention, wherein the size of the excipient nano-domain is from about 100 nm to about 200 nm. [The present invention 1016] Any of the pharmaceutical compositions according to claims 1001 to 1015, having a mass median aerodynamic particle diameter of about 1.5 to about 7.5 µm. [The present invention 1017] The pharmaceutical composition of the present invention, wherein the mass median aerodynamic particle diameter is about 2.5 to about 6.5 μm. [The present invention 1018] The pharmaceutical composition of any one of claims 1001 to 1017, which does not contain a wax excipient. [The present invention 1019] A pharmaceutical composition according to the present invention, which does not contain a hydrophobic excipient. [The present invention 1020] The therapeutic agent is selected from the group consisting of anticancer agents, antifungal agents, psychoneurological agents such as analgesics, agents that alter the level of consciousness such as anesthetics or hypnotics, nonsteroidal anti-inflammatory drugs (NSAIDS), anthelmintics, beta agonists, antiacne agents, antiangina pectoris agents, antiarrhythmic agents, antiasthma agents, antimicrobial agents, antibenign prostatic hyperplasia agents, anticoagulants, antidepressants, antidiabetic agents, antiemetic agents, antiepileptic agents, antigout agents, antihypertensive agents, antiinflammatory agents, antimalarial agents, antimigraine agents, antimuscarinic agents, antitumor agents, antiobesity agents, antiosteoporosis agents, antiparkinsonian agents, antiproliferative agents, antiprotozoal agents, The pharmaceutical composition of any of the present inventions 1001 to 1019, wherein the pharmaceutical composition is selected from the group including antithyroid agents, antitussives, anti-urinary incontinence agents, antivirals, anti-anxieties, appetite suppressants, beta blockers, cardiac inotropes, chemotherapeutics, cognition enhancers, contraceptives, corticosteroids, Cox-2 inhibitors, diuretics, erectile dysfunction ameliorators, expectorants, gastrointestinal agents, histamine receptor antagonists, immunosuppressants, keratolytics, lipid regulating agents, leukotriene inhibitors, macrolides, muscle relaxants, neuroleptics, nutritional agents, opioid analgesics, protease inhibitors, and sedatives. [The present invention 1021] The pharmaceutical composition of the present invention, wherein the therapeutic agent is an antifungal agent. [The present invention 1022] The pharmaceutical composition of the present invention, wherein the antifungal agent is an azole antifungal agent. [The present invention 1023] The pharmaceutical composition of the present invention, wherein the azole antifungal drug is voriconazole. [The present invention 1024] The pharmaceutical composition of any of claims 1001 to 1023, further comprising one or more additional excipients. [The present invention 1025] The pharmaceutical composition of any of claims 1001 to 1024, further comprising one or more additional therapeutic agents. [The present invention 1026] The pharmaceutical composition of any one of the present inventions 1001 to 1025, which is formulated for administration orally, intraadiposely, intra-arterially, intra-articularly, intracranially, intradermally, intralesionally, intramuscularly, intranasally, intraocularly, intrapericardially, intraperitoneally, intrapleurally, intraprostatically, intrarectally, intrathecally, intratracheally, intratumorally, intraumbilically, intravaginally, intravenously, intravesically, intravitreally, in liposomes, partially, mucosally, parenterally, rectally, subconjunctivally, subcutaneously, sublingually, topically, transdermally, vaginally, in the form of a cream, in the form of a lipid composition, by catheter, by lavage, by continuous infusion, by injection, by inhalation, by injection, by local delivery, or by local perfusion. [The present invention 1027] A pharmaceutical composition of the present invention 1026 which is formulated for administration by inhalation. [The present invention 1028] The pharmaceutical composition of any one of claims 1001 to 1027, which is formulated for use in an inhaler. [The present invention 1029] The pharmaceutical composition of the present invention 1028, wherein said inhaler is a fixed dose combination inhaler, a single dose dry powder inhaler, a multiple dose dry powder inhaler, a multiple unit dose dry powder inhaler, a metered dose inhaler, or a pressurized metered dose inhaler. [The present invention 1030] The pharmaceutical composition of the present invention, wherein said inhaler is a capsule-based inhaler. [The present invention 1031] The pharmaceutical composition of any one of claims 1028 to 1030, wherein the inhaler is a low resistance inhaler. [The present invention 1032] The pharmaceutical composition of any one of claims 1028 to 1030, wherein the inhaler is a high resistance inhaler. [The present invention 1033] The pharmaceutical composition of any one of claims 1028 to 1032, wherein the inhaler is used at a flow rate of about 10 L / min to about 150 L / min. [The present invention 1034] The pharmaceutical composition of the present invention, wherein the flow rate is about 20 L / min to about 100 L / min. [The present invention 1035] The pharmaceutical composition of any of claims 1028 to 1034, wherein the inhaler has a pressure difference of 0.5 kPa to about 5 kPa. [The present invention 1036] The pharmaceutical composition of the present invention 1035, wherein the pressure difference is 1 kPa, 2 kPa, or 4 kPa. [The present invention 1037] 1037. The pharmaceutical composition of any one of claims 1028 to 1036, wherein the inhaler has a loading dose of about 0.1 mg to about 50 mg. [The present invention 1038] The pharmaceutical composition of the present invention 1037, wherein said inhaler has a loading dose of about 0.1 mg to about 10 mg. [The present invention 1039] The pharmaceutical composition of the present invention 1037, wherein said inhaler has a loading dose of about 5 mg to about 50 mg. [The present invention 1040] The pharmaceutical composition of the present invention, wherein the loading dose is about 5 mg to about 25 mg. [The present invention 1041] The pharmaceutical composition of any of claims 1001-1040, wherein the inhaler is configured to deliver one or a series of doses from one or more unit doses loaded in series. [The present invention 1042] The pharmaceutical composition of claim 1041, wherein said inhaler is configured to deliver one dose from one unit dose. [The present invention 1043] The pharmaceutical composition of claim 1041, wherein the inhaler is configured to deliver a series of doses from one unit dose. [The present invention 1044] 1041. The pharmaceutical composition of claim 1041, wherein the inhaler is configured to deliver one dose each from a series of successively loaded capsules. [The present invention 1045] 1041. The pharmaceutical composition of claim 1041, wherein the inhaler is configured to deliver a series of doses from a series of capsules loaded in series. [The present invention 1046] A method for treating or preventing a disease or disorder in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of any of the pharmaceutical compositions of present inventions 1001 to 1045, which comprises a therapeutic agent effective to treat the disease or disorder. [The present invention 1047] The method of claim 1046, wherein the disease or disorder is in the lungs. [The present invention 1048] The method of claim 1046 or 1047, wherein the disease or disorder is an infectious disease. [The present invention 1049] The method according to any one of claims 1046 to 1048, wherein the infectious disease is caused by a fungus. [The present invention 1050] The method of claim 1049, wherein said therapeutic agent is an antifungal agent. [The present invention 1051] The method of claim 1050, wherein said therapeutic agent is an azole antifungal agent. [The present invention 1052] The method of claim 1051, wherein said therapeutic agent is voriconazole. [The present invention 1053] 1. A method for preparing a pharmaceutical composition comprising the steps of: (A) mixing a therapeutic agent and excipients present in an amount less than 10% w / w with a solvent to form a precursor solution; (B) depositing the precursor solution onto a surface at a temperature suitable to freeze the solvent; and (C) Removing the solvent to obtain a pharmaceutical composition. The method comprising: [The present invention 1054] The process of claim 1053, wherein the solvent is a mixture of two or more solvents. [The present invention 1055] The process of claim 1054, wherein said mixture of solvents comprises water. [The present invention 1056] The method of claim 1055, wherein the solvent is an organic solvent. [The present invention 1057] The process of claim 1056, wherein the organic solvent is acetonitrile. [The present invention 1058] The process of claim 1056, wherein the organic solvent is 1,4-dioxane. [The present invention 1059] The method according to any one of claims 1053 to 1057, wherein the solvent is a mixture of water and an organic solvent. [The present invention 1060] 1059. The process of claim 1059, wherein the solvent is a mixture of water and acetonitrile. [The present invention 1061] The method of any of claims 1053 to 1060, wherein said mixture of two or more solvents comprises from about 10% v / v to about 90% v / v of an organic solvent. [The present invention 1062] The method of claim 1061, wherein the mixture comprises about 40% v / v to about 60% v / v of the organic solvent. [The present invention 1063] The method of claim 1062, wherein the mixture comprises about 50% v / v of the organic solvent. [The present invention 1064] The method of claim 1061, wherein the mixture comprises about 20% v / v to about 40% v / v of the organic solvent. [The present invention 1065] The method of claim 1064, wherein the mixture comprises about 30% v / v of the organic solvent. [The present invention 1066] The method of any one of claims 1053 to 1065, wherein said therapeutic agent and excipients constitute less than 10% w / v of said precursor solution. [The present invention 1067] The method of claim 1066, wherein said therapeutic agent and excipients comprise from about 0.5% to about 5% w / v of said precursor solution. [The present invention 1068] The method of claim 1067, wherein said therapeutic agent and excipients comprise about 1% w / v of said precursor solution. [The present invention 1069] The method of claim 1067, wherein said therapeutic agent and excipients comprise about 3% w / v of said precursor solution. [The present invention 1070] Any of the methods of claims 1053 to 1069, wherein the surface is rotating. [The present invention 1071] The method according to any one of claims 1053 to 1070, wherein the temperature is from about 0°C to about -200°C. [The present invention 1072] The method of claim 1071, wherein the temperature is from about 0°C to about -120°C. [The present invention 1073] The method of claim 1072, wherein the temperature is from about -50°C to about -90°C. [The present invention 1074] The method of claim 1073, wherein the temperature is about -60°C. [The present invention 1075] The method of claim 1072, wherein the temperature is from about -125°C to about -175°C. [The present invention 1076] The method of claim 1073, wherein the temperature is about -150°C. [The present invention 1077] The process of any one of claims 1053 to 1076, wherein the solvent is removed under reduced pressure. [The present invention 1078] The process of claim 1077, wherein the solvent is removed by lyophilization. [The present invention 1079] The method according to claim 1078, wherein said freeze-drying is carried out at a freeze-drying temperature of about -20°C to about -100°C. [The present invention 1080] The method of claim 1079, wherein the freeze-drying temperature is about -40°C. [The present invention 1081] The method according to any one of claims 1077 to 1080, wherein the reduced pressure is less than 250 mTorr. [The present invention 1082] The method of claim 1081, wherein the reduced pressure is about 100 mTorr. [The present invention 1083] The method of any of claims 1053 to 1082, further comprising heating the pharmaceutical composition under reduced pressure. [The present invention 1084] The method of claim 1083, wherein the pharmaceutical composition is heated to a temperature of about 0°C to about 30°C. [The present invention 1085] The method of claim 1084, wherein said temperature is at or near room temperature or about 25°C. [The present invention 1086] The method according to any one of claims 1083 to 1085, wherein the reduced pressure is less than 250 mTorr. [The present invention 1087] The method of claim 1086, wherein the reduced pressure is about 100 mTorr. [The present invention 1088] The method of any of claims 1083 to 1087, wherein the reduced pressure is the same as that during freeze-drying. [The present invention 1089] A pharmaceutical composition prepared according to any one of the methods of the present invention. Other objects, features, and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that this detailed description and the specific examples, while indicating specific embodiments of the present invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description. [Brief description of the drawings]

[0020] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure. The present disclosure may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0021] [Figure 1] Figure 1 shows 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. [Diagram 2] Modulated DSC of (a) TFF-MAN, (b) TFF-VCZ, (c) TFF-VCZ-MAN 95:5, and (d) TFF-VCZ-MAN 50:50. [Figure 3-1]SEM images of TFF-VCZ-MAN are shown: (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 3-2] Shown are SEM images of TFF-VCZ-MAN: (Figure 3G) aerosolized TFF-VCZ-MAN 95:5; (Figure 3H) aerosolized TFF-VCZ-MAN 50:50; (Figure 3I) TFF-VCZ-MAN 25:75 after 5 min in a Franz cell; (Figure 3J) TFF-VCZ-MAN 95:5 after 5 min in a Franz cell. [Figure 4] (Figure 4A) SEM image of TFF-VCZ, (Figure 4B) SEM image of TFF-VCZ-MAN 95:5, (Figure 4C) 3D topography image of TFF-VCZ, (Figure 4D) 3D topography image of TFF-VCZ-MAN 95:5, (Figure 4E) diagram of contact area and distance between particles of TFF-VCZ, and (Figure 4F) diagram of contact area and distance between particles of TFF-VCZ-MAN 95:5. [Diagram 5] 13 shows an AFM topography image of TFF-VCZ-MAN 95:5 aerosolized with a DP4 insufflator. [Figure 6] The SSA of the TFF-VCZ-MAN powder formulation is shown (n=3; mean±SD). [Figure 7] Shows SEM / EDX data of TFF-VCZ-MAN 50:50: (Figure 7A) SEM image; (Figure 7B) elemental analysis of spot A; (Figure 7C) elemental analysis of spot B. [Figure 8] FT-IR (Figure 8A, region 3500 cm-1 to 3100 cm-1; Figure 8B, region 1290 cm-1 to 1230 cm-1) 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. [Figure 9](Figure 9A) 1D CP-MAS spectrum of TFF-VCZ, (Figure 9B) 1D CP-MAS spectrum of TFF-VCZ-MAN 90:10, showing the 13C spectrum (left spectrum) and the 19F spectrum (right spectrum). [Figure 10] (Figure 10A) 2D 1H-13C HETCOR spectrum of TFF-VCZ, (Figure 10B) 2D 1H-13C HETCOR spectrum of TFF-VCZ-MAN 90:10. [Figure 11] The FPF (% of weighed value) of the TFF-VCZ-MAN dry powder formulations is shown (n=3, mean±SD). [Figure 12] Aerodynamic particle size distribution profiles of TFF-VCZ-MAN 95:5 for shear times of 0, 15, 30, and 60 min (from left to right) are shown (n=3; mean ± SD). [Figure 13] Aerodynamic properties of TFF-VCZ-MAN 95:5 versus shear time: (line a) FPF, % of delivered dose; (line b) FPF, % of metric dose; (line c) MMAD; (line d) GSD (n=3; mean ± SD). [Figure 14] Aerodynamic properties over time of TFF-VCZ-MAN 95:5 stored at 25 °C / 60% RH: (line a) FPF, % of delivered dose; (line b) FPF, % of metric dose; (line c) MMAD; (line d) GSD (n=3; mean ± SD). [Figure 15] Cumulative voriconazole release (%) is shown for (a) TFF-VCZ-PVPK25 25:75 (amorphous), (b) TFF-VCZ-MAN 25:75, (c) TFF-VCZ-MAN 50:50, and (d) TFF-VCZ-MAN 95:5 (n=3; mean ± SD). [Figure 16] Images of the freezing process are shown. [Figure 17] (a) AFM topography image of formulation #2 (scale 5 μm×5 μm), (b) AFM topography image of formulation #4 (scale 2 μm×2 μm), (c) corresponding 3D topography image of formulation #2, (d) corresponding 3D topography image of formulation #4. [Figure 18]1 shows SEM images of voriconazole nanoaggregates for (a) Formulation #1, (b) Formulation #2, (c) Formulation #3, (d) Formulation #4, (e) Formulation #5, and (f) Formulation #6. [Figure 19] 1 shows SEM images of aerosolized voriconazole nanoaggregates for (a)-(b) Formulation #7, (c)-(f) Formulation #6. [Figure 20] 1 shows the XRPD of (a) voriconazole powder, (b) TFF-voriconazole, (c) formulation #6 (small scale), (d) formulation #6 (large scale), and (e) TFF-mannitol. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] Description of exemplary embodiments In some aspects of the present disclosure, pharmaceutical compositions include nanoaggregates. These compositions can be prepared by methods such as thin film freezing and include a therapeutic agent and an excipient. In some embodiments, these compositions also exhibit improved aerosolization or provide other pharmaceutical properties.

[0023] Methods for preparing and using these compositions are also provided herein, the details of which are set forth in more detail below.

[0024] I. Pharmaceutical Compositions In some embodiments, 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 include freezing a solvent solution of the therapeutic agent and excipients, and then removing the solvent either at reduced pressure and / or at low temperature. Methods of 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, Beinborn et al., 2012a, Beinborn et al., 2012b, 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. Pat. No. 8,968,786, all of which are incorporated herein by reference.

[0025] Such pharmaceutical compositions may exist as nanoaggregates, which include collections of nanoparticles attracted or bound by intermolecular or intramolecular cohesive forces. In the pharmaceutical compositions described herein, the nanoaggregates may include one or more particles of drug coated with distinct discontinuous nanodomains of excipient. Without wishing to be bound by any theory, the nanodomains of excipient may include sizes of 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 any range of sizes derivable therein. The size of these nano-domains of the excipients includes sizes 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 separate compositions interspersed on the surface of the nano-aggregates containing the therapeutic agent. The pharmaceutical composition may further include 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, 2, 75, 3.0, 3.25, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.25, 5.5, 5.75, 6.0, 6.25, 6.5, 6.75, 7.0, 7.25 to about 7.5 μm, or any range derivable therefrom.

[0026] A.Medical Drugs A "therapeutic agent" as used in the methods and compositions of the invention refers to any substance, compound, drug, agent, or other major active ingredient that provides a therapeutic or pharmacological effect when administered to a human or animal. When a therapeutic agent is present in the composition, the therapeutic agent is present in the composition at a level of about 50% to about 99% w / w, about 70% to about 99% w / w, about 90% to about 97% w / w, or about 95% to about 97% w / w of the total composition. In some embodiments, the amount of therapeutic agent is about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% to about 99% w / w, or any range derivable therein.

[0027] Suitable lipophilic therapeutic agents may be any poorly water soluble biologically active drug or a salt, isomer, ester, ether or other derivative thereof, including, but not limited to, anti-cancer drugs, anti-fungal drugs, psycho-neurological drugs such as analgesics, drugs that alter the level of consciousness such as anesthetics or hypnotics, non-steroidal anti-inflammatory drugs (NSAIDS), anthelmintics, anti-acne drugs, anti-angina drugs, anti-arrhythmic drugs, anti-asthma drugs, antibacterial drugs, anti-benign prostatic hyperplasia drugs, anticoagulants, antidepressants, antidiabetic drugs, antiemetic drugs, antiepileptic drugs, antigout drugs, antihypertensive drugs, anti-inflammatory drugs, antimalarial drugs, antimigraine drugs, antimuscarinic drugs, anti-tumor drugs, The therapeutic agent may include, but is not limited to, an anti-cancer agent, an anti-obesity agent, an anti-osteoporosis agent, an anti-Parkinson's agent, an anti-proliferative agent, an antiprotozoal agent, an antithyroid agent, an antitussive, an anti-urinary incontinence agent, an antiviral agent, an anxiolytic, an appetite suppressant, a beta agonist, a beta blocker, a cardiac inotropic agent, a chemotherapy agent, a cognitive enhancer, a contraceptive, a corticosteroid, a Cox-2 inhibitor, a diuretic, an erectile dysfunction agent, an expectorant, a gastrointestinal agent, a histamine receptor antagonist, an immunosuppressant, a keratolytic agent, a lipid regulating agent, a leukotriene inhibitor, a macrolide, a muscle relaxant, a neuroleptic, a nutritional agent, an opioid analgesic, a protease inhibitor, or a sedative.

[0028] Non-limiting examples of therapeutic agents include 7-methoxypteridine, 7-methylpteridine, abacavir, abafungin, abarelix, acebutolol, acenaphthene, acetaminophen, acetanilide, acetazolamide, acetohexamide, acetretin, acrivastine, adenine, adenosine, alatrofloxacin, albendazole, albuterol, alclofenac, aldesleukin, alemtuzumab, alfuzosin, alitretinoin, allobarbital, allopurinol, all-trans retinoic acid (ATRA), aloxipirin, alprazolam, alprenolo, and the like. , altretamine, amifostine, amiloride, aminoglutethimide, aminopyrine, amiodarone HCl, amitriptyline, amlodipine, amobarbital, amodiaquine, amoxapine, amphetamine, amphotericin, amphotericin B, ampicillin, amprenavir, amsacrine, amyl nitrate, amylobarbitone, anastrozole, amrinone, anthracene, anthracycline, aprobarbital, arsenic trioxide, asparaginase, aspirin, astemizole, atenolol, atorvastatin, atovaquone, atrazine, atropine, atropine Azathioprine, Auranofin, Azacitidine, Azapropazone, Azathioprine, Azintamide, Azithromycin, Aztreonam, Baclofen, Barbitone, BCG live bacteria, Beclamide, Beclomethasone, Bendroflumethiazide, Benezepryl, Benidipine, Benorilate, Benperidol, Bentazepam, Benzamide, Benzanthracene, Benzathine penicillin, Benzhexol HCl, Benznidazole, Benzodiazepine, Benzoic acid, Hydroxynaphthoic acid, Bephenium, Betamethasone, Bevacizine Mab (Avastin), Bexarotene, Bezafibrate, Bicalutamide, Bifonazole, Biperiden, Bisacodyl, Bisantrene, Bleomycin, Bortezomib, Brinzolamide, Bromazepam, Bromocriptine Mesylate, Bromperidol, Brotizolam, Budesonide, Bumetanide, Bupropion, Busulfan, Butalbital, Butamben, Butenafine HCl, Butobarbitone, Butobarbitone (Butetal), Butoconazole, Butoconazole Nitrate, Butylparaben, Caffeine,calcifediol, calciprotriene, calcitriol, calsterone, campbendazole, camphor, camptothecin, camptothecin analogues, candesartan, capecitabine, capsaicin, captopril, carbamazepine, carbimazole, carbofuran, carboplatin, carbromal, carimazole, carmustine, cefamandole, cefazolin, cefixime, ceftazidime, cefuroxime axetil, celecoxib, cephradine, cerivastatin, cetirizine, cetuximab, chlorambucil, chloram Phenicol, Chlordiazepoxide, Clomethiazole, Chloroquine, Chlorothiazide, Chlorpheniramine, Chlorproguanil HCl, Chlorpromazine, Chlorpropamide, Chlorprothixene, Chlorpyrifos, Chlortetracycline, Chlorthalidone, Chlorzoxazone, Cholecalciferol, Chrysene, Cilostazol, Cimetidine, Cinnarizine, Cinoxacin, Ciprofibrate, Ciprofloxacin HCl, Cisapride, Cisplatin, Citalopram, Cladribine, Clarithromycin, Clemastine Fumarate, Clioquinoline aztreonam, clobazam, clofarabine, clofazimine, clofibrate, clomiphene citrate, clomipramine, clonazepam, clopidogrel, clotiazepam, clotrimazole, cloxacillin, clozapine, cocaine, codeine, colchicine, colistin, conjugated estrogens, corticosterone, cortisone, cortisone acetate, cyclizine, cyclobarbital, cyclobenzaprine, cyclobutane-spirobarbiturate, cycloethane-spirobarbiturate, cycloheptane-spirobarbiturate, cyclohexene ... San-spirobarbiturate, cyclopentane-spirobarbiturate, cyclophosphamide, cyclopropane-spirobarbiturate, cycloserine, cyclosporine, cyproheptadine, cyproheptadine HCl, cytarabine, cytosine, dacarbazine, dactinomycin, danazol, danthron, dantrolene sodium, dapsone, darbepoetin alfa, dalodipine, daunorubicin, decoquinate, dehydroepiandrosterone, delavirdine, demeclocycline, denileukin, deoxycorticosterone, desoxymetasone,Dexamethasone, dexamphetamine, dexchlorpheniramine, dexfenfluramine, dexrazoxane, dextropropoxyphene, diamorphine, diatrizoate, diazepam, diazoxide, dichlorophene, dichlorprop, diclofenac, dicumarol, didanosine, diflunisal, digitoxin, digoxin, dihydrocodeine, dihydroequilene, dihydroergotamine mesylate, diiodohydroxyquinoline, diltiazem HCl, diloxanide furoate, dimenhydrinate, dimorpholamine, dinitramide, Diosgenin, Diphenoxylate HCl, Diphenyl, Dipyridamole, Dirithromycin, Disopyramide, Disulfiram, Diuron, Docetaxel, Domperidone, Donepezil, Doxazosin, Doxazosin HCl, Doxorubicin (neutral), Doxorubicin HCl, Doxycycline, Dromostanolone Propionate, Droperidol, Dyphylline, Echinocandin, Econazole, Econazole Nitrate, Efavirenz, Ellipticine, Enalapril, Enlimomab, Enoximone, Epinephrine, Epipodophyllotoxin Derivatives, Epil Bicine, epoetin alfa, eposartan, equilenin, equilin, ergocalciferol, ergotamine tartrate, erlotinib, erythromycin, estradiol, estramustine, estriol, estrone, ethacrynic acid, ethambutol, etinamate, ethionamide, ethopropazine HCl, ethyl-4-aminobenzoate (benzocaine), ethylparaben, ethinyl estradiol, etodolac, etomidate, etoposide, etretinate, exemestane, felbamate, felodipine, fenbendazole, fen embuconazole, fenbufen, fenchlorphos, fenclofenac, fenfluramine, fenofibrate, fenoldepam, fenoprofen calcium, fenoxycarb, fenpiclonil, fentanyl, fenticonazole, fexofenadine, filgrastim, finasteride, flecamid acetate, floxuridine, fludarabine, fluconazole, flucytosine, fludioxonil, fludrocortisone, fludrocortisone acetate, flufenamic acid, fluanisone, flunarizine HCl, flunisolide,Flunitrazepam, fluocortolone, fluometuron, fluorene, fluorouracil, fluoxetine HCl, fluoxymesterone, flupentixol decanoate, fluphentixol decanoate, flurazepam, flurbiprofen, fluticasone propionate, fluvastatin, folic acid, fosenopril, fosphenytoin sodium, frovatriptan, furosemide, fulvestrant, furazolidone, gabapentin, G-BHC (lindane), gefitinib, gemcitabine, gemfibrozil, gemtuzumab, glafenine, glibenc Lamide, gliclazide, glimepiride, glipizide, glutethimide, glyburide, glyceryl trinitrate (nitroglycerin), goserelin acetate, grepafloxacin, griseofulvin, guaifenesin, guanabenzacetate, guanine, halofantrine HCl, haloperidol, hydrochlorothiazide, heptabarbital, heroin, hesperetin, hexachlorobenzene, hexethal, histrelin acetate, hydrocortisone, hydroflumethiazide, hydroxyurea, hyoscyamine, hypoxanthine, ibritumomab, ibuprofen Fen, idarubicin, idobutal, ifosfamide, dihydroequilenin, imatinib mesylate, imipenem, indapamide, indinavir, indomethacin, indoprofen, interferon alpha-2a, interferon alpha-2b, iodamide, iopanoic acid, iprodione, irbesartan, irinotecan, isabuconazole, isocarboxazid, isoconazole, isoguanine, isoniazid, isopropyl barbiturate, isoproturon, isosorbide dinitrate, isosorbide mononitrate, isradipine , itraconazole, itraconazole, itraconazole (Itra), ivermectin, ketoconazole, ketoprofen, ketorolac, khellin, labetalol, lamivudine, lamotrigine, lanatoside C, lanospragole, L-DOPA, leflunomide, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, levofloxacin, lidocaine, linuron, lisinopril, lomefloxacin, lomustine, loperamide, loratadine, lorazepam, lorefloxacin, lormetazepam, losartan mesylate, lovastatin,Lisuride maleate, Maprotiline HCl, Mazindol, Mebendazole, Meclizine HCl, Meclofenamic acid, Medazepam, Medigoxin, Medroxyprogesterone acetate, Mefenamic acid, Mefloquine HCl, Megestrol acetate, Melphalan, Mepenzolate bromide, Meprobamate, Meptazinol, Mercaptopurine, Mesalazine, Mesna, Mesoridazine, Mestranol, Methadone, Methaqualone, Methocarbamol, Methoine, Methotrexate, Methoxsalen, Methsuximide, Methyclothiazide, Methylphenidate, Methylphenidate Novarbitone, Methyl-p-hydroxybenzoate, Methylprednisolone, Methyltestosterone, Methyprilon, Methysergide Maleate, Metoclopramide, Metolazone, Metoprolol, Metronidazole, Mianserin HCl, Miconazole, Midazolam, Mifepristone, Miglitol, Minocycline, Minoxidil, Mitomycin C, Mitotane, Mitoxantrone, Mofetil Mycophenolate, Molindone, Montelukast, Morphine, Moxifloxacin HCl, Nabumetone, Nadolol, Nalbuphine, Nalidixic Acid, Nandrolone lon, naphthacene, naphthalene, naproxen, naratriptan HCl, natamycin, nelarabine, nelfinavir, nevirapine, nicardipine HCl, nicotinamide, nicotinic acid, nicoumarone, nifedipine, nilutamide, nimodipine, nimorazole, nisoldipine, nitrazepam, nitrofurantoin, nitrofurazone, nizatidine, nofetumomab, norethisterone, norfloxacin, norgestrel, nortriptyline HCl, nystatin, estradiol, ofloxacin, olanzapine, omeprazole, omoconazole ... Andansetron HCl, Oprelvequin, Ornidazole, Oxaliplatin, Oxamniquine, Oxanterembonate, Oxaprozin, Oxatomide, Oxazepam, Oxcarbazepine, Oxifendazole, Oxiconazole, Oxprenolol, Oxyphenbutazone, Oxyphencyclimine HCl, Paclitaxel, Palifermin, Pamidronate, p-aminosalicylic acid, Pantoprazole, Paramethadione, Paroxetine HCl, Pegademase, Pegaspargase, Pegfilgrastim, Pemetrexed disodium,Penicillamine, pentaerythritol tetranitrate, pentazocine, pentazocine, pentobarbital, pentobarbitone, pentostatin, pentoxifylline, perphenazine, perphenazine pimozide, perylene, phenacemide, phenacetin, phenanthrene, phenindione, phenobarbital, f, Phenol barbitone, phenolphthalein, phenoxybenzamine, phenoxybenzamine HCl, phenoxymethylpenicillin, phensuximide, phenylbutazone, phenytoin, pindolol, pioglitazone, pipobroman, piroxicam, pizotifen maleate, platinum compounds, plicamycin, polyenes, polymyxin B, porfimer sodium, posaconazole (Posa), pramipexole, prasterone, pravastatin, praziquantel, prazosin, prazosin HCl, prednisolone, prednisone, primidone, proba lubital, probenecid, probucol, procarbazine, prochlorperazine, progesterone, proguanil HCl, promethazine, propofol, propoxar, propranolol, propylparaben, propylthiouracil, prostaglandins, pseudoephedrine, pteridine-2-methyl-thiol, pteridine-2-thiol, pteridine-4-methyl-thiol, pteridine-4-thiol, pteridine-7-methyl-thiol, pteridine-7-thiol, pyranterembonate, pyrazinamide, pyrene, pyridostigmine, Pyrimethamine, quetiapine, quinacrine, quinapril, quinidine, quinidine sulfate, quinine, quinine sulfate, rabeprazole sodium, ranitidine HCl, rasburicase, ravuconazole, repaglinide, reposar, reserpine, retinoids, rifabutin, rifampicin, rifapentine, rimexolone, risperidone, ritonavir, rituximab, rizatriptan benzoate, rofecoxib, ropinirole HCl, rosiglitazone, saccharin, salbutamol, salicylamide, salicylic acid, saquinavir, sargramostim, secbuta Barbiturate, Secobarbital, Sertaconazole, Sertindole, Sertraline HCl, Simvastatin, Sirolimus, Sorafenib, Sparfloxacin, Spiramycin, Spironolactone, Stanolone, Stanozolol, Stavudine, Stilbestrol, Streptozocin, Strychnine, Sulconazole, Sulconazole Nitrate, Sulfacetamide, Sulfadiazine, Sulfamerazine, Sulfamethazine, Sulfamethoxazole, Sulfanilamide, Sulfathiazole, Sulindac, Sulfabenzamide, Sulfacetamide,Sulfadiazine, sulfadoxine, sulfafurazole, sulfamerazine, sulfamethoxazole, sulfapyridine, sulfasalazine, sulfinpyrazone, sulpiride, sulthiame, sumatriptan succinate, sunitinib maleate, tacrine, tacrolimus, talbutal, tamoxifen citrate, tamsulosin, targretin, taxanes, tazarotene, telmisartan, temazepam, temozolomide, teniposide, tenoxicam, teratoma. Zosyn, Terazosin HCl, Terbinafine HCl, Terbutaline Sulfate, Terconazole, Terfenadine, Testolactone, Testosterone, Tetracycline, Tetrahydrocannabinol, Tetroxoprim, Thalidomide, Thebaine, Theobromine, Theophylline, Thiabendazole, Thiamphenicol, Thioguanine, Thioridazine, Thiotepa, Thotoin, Thymine, Tiagabine HCl, Tibolone, Ticlopidine, Tinidazole, Tioconazo ol, tirofiban, tizanidine HCl, tolazamide, tolbutamide, tolcapone, topiramate, topotecan, toremifene, tositumomab, tramadol, trastuzumab, trazodone HCl, tretinoin, triamcinolone, triamterene, triazolam, triazole, triflupromazine, trimethoprim, trimipramine maleate, triphenylene, troglitazone, tromethamine, tropicamide, trovafloxacin, chibamate, Included may be mentioned are ubidecarenone (coenzyme Q10), undecenoic acid, uracil, uracil mustard, uric acid, valproic acid, valrubicin, valsartan, vancomycin, venlafaxine HCl, vigabatrin, vinbarbital, vinblastine, vincristine, vinorelbine, voriconazole, xanthine, zafirlukast, zidovudine, zileuton, zoledronate, zoledronic acid, zolmitriptan, zolpidem, and zopiclone.

[0029] In certain embodiments, the therapeutic agent can be voriconazole or other members of the general azole compound group.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, and terconazole; and c) thiazoles, such as abafungin.Other drugs that can be used in this approach include, but are not limited to, hyperthyroidism drugs, such as kalimazole; anticancer drugs, such as cytotoxic agents, such as epipodophyllotoxin derivatives, taxanes, bleomycin, anthracyclines, and platinum compounds and camptothecin analogues. The following therapeutic agents may also include other antifungal antibiotics, such as poorly water-soluble echinocandins, polyenes (e.g., amphotericin B and natamycin), as well as antibacterial agents (e.g., polymyxin B and colistin) and antiviral agents. These drugs may also include psychotropic agents, such as antipsychotics, antidepressants, or analgesics and / or sedatives, such as benzodiazepines. These drugs may also include agents that alter the level of consciousness or anesthetics, such as propofol. The present compositions and methods for making them can be used to prepare pharmaceutical compositions having suitable pharmacokinetic properties for use as therapeutic agents.

[0030] In some embodiments, the compositions described herein may include long-acting beta agonists (LABA).Some non-limiting examples of long-acting beta agonists include formoterol, such as formoterol fumarate, salmeterol, such as salmeterol xinafoate, bambuterol, clenbuterol, indacaterol, olodaterol, protokylol, avesiterol, salmefamol, vilanterol, arformoterol, carmoterol, PF-610355, GSK-159797, GSK-597901, GSK-159802, GSK-642444, GSK-678007, or other long-acting beta agonists known in the art.

[0031] In other embodiments, the compositions described herein may include long-acting muscarinic antagonists (LAMA).Some non-limiting examples of long-acting muscarinic antagonists include tiotropium, aclidinium, dexpyronium, ipratropium, oxitropium, darotropium, glycopyrronium, or salts of glycopyrrolate derivatives, or other long-acting muscarinic antagonists known in the art, such as those taught in U.S. Patent Application No. 2009 / 0181935, PCT Patent Application No. WO2010 / 007561, and PCT Patent Application No. WO2008 / 035157, which are incorporated herein by reference.

[0032] In other embodiments, the compositions described herein may include a corticosteroid, particularly a corticosteroid suitable for inhalation, some non-limiting examples of which include beclomethasone dipropionate, budesonide, flunisolide, fluticasone propionate, fluticasone furoate, mometasone furoate, ciclesonide, rofleponide palmitate, triamcinolone acetonide, or other corticosteroids known in the art.

[0033] In other embodiments, the compositions described herein may include one or more antibiotic agents. Some types of antibiotics include penicillins, cephalosporins, carbapenems, macrolides, aminoglycosides, quinolones (including fluoroquinolones), sulfonamides, and tetracyclines. In some embodiments, the compositions may include narrow-spectrum antibiotics that target specific bacterial species. Some non-limiting examples of bactericidal antibiotics include penicillins, cephalosporins, polymyxins, rifamycins, lipiarmycins, quinolones, and sulfonamides. Some non-limiting examples of bacteriostatic antibiotics include macrolides, lincosamides, or tetracyclines. In some embodiments, the antibiotic is an aminoglycoside, such as kanamycin and streptomycin, ansamycin, such as rifaximin and geldanamycin, carbacephems, 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, monobactams, such as For example, aztreonam, nitrofurans such as furazolidone and nitrofurantoin, oxazolidonones 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 sulfasalazine, or tetracyclines such as demeclocycline, doxycycline, minocycline, oxytetracycline, or tetracycline.In some embodiments, the composition includes drugs that act against mycobacteria, such as cycloserine, capreomycin, ethionamide, rifampicin, rifabutin, rifapentine, and streptomycin. Other antibiotics considered may include arsphenamine, chloramphenicol, fosfomycin, fusidic acid, metronidazole, mupirocin, platensimycin, quinupristin, dalfopristin, thiamphenicol, tigecycline, tinidazole, or trimethoprim.

[0034] 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, azole antifungal compounds, echinocandins, or flucytosine.Some non-limiting examples of azole antifungal compounds 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.

[0035] In some embodiments, the compositions may further comprise one or more antiviral agents, such as nucleoside analogues, such as acyclovir, famciclovir, valacyclovir, penciclovir, and ganciclovir, or other antiviral agents, such as pegylated interferon, interferon alpha-2b, lamivudine, adefovir, telbivudine, entecavir, or tenofovir.

[0036] B. Excipients In some embodiments, the present disclosure includes one or more excipients that are incorporated into the pharmaceutical composition. In some embodiments, the excipients used herein are water-soluble excipients. These water-soluble excipients include sugars, 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 some embodiments, these excipients are solid at room temperature. Some non-limiting examples of sugar alcohols include erythritol, threitol, arabitol, xylitol, ribitol, mannitol, sorbitol, galactitol, fucitol, iditol, inositol, volemitol, isomalt, maltitol, lactitol, maltotritol, maltotetraitol, or polyglycitol. In some embodiments, the pharmaceutical composition of the present invention may further be free of hydrophobic or waxy excipients, such as waxes and oils. Some non-limiting examples of hydrophobic excipients include hydrogenated and partially hydrogenated oils, palm oil, soybean oil, castor oil, carnauba wax, beeswax, palm wax, white wax, castor oil wax, or lanolin.In addition, the present disclosure may further include one or more amino acids, or their amide or ester derivatives.In some embodiments, the amino acid used may be one of the 20 standard 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 may be in D or L configuration, or the amino acids may be α-, β-, γ-, or δ-amino acids.In other embodiments, one of the common non-standard amino acids may be used, such as carnitine, GABA, carboxyglutamic acid, levothyroxine, hydroxyproline, selenomethionine, beta-alanine, ornithine, citrulline, dehydroalanine, δ-aminolevulinic acid, or 2-aminoisobutyric acid.

[0037] In some embodiments, the amount of excipient in the pharmaceutical composition is about 0.5% to about 10% w / w, about 1% to about 10% w / w, about 2% to about 8% w / w, or about 2% to about 5% w / w. The amount of excipient in the pharmaceutical composition is about 0.5%, 0.75%, 1%, 1.25%, 1.5%, 1.75%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, 8%, 9% to about 10% w / w of the total pharmaceutical composition, or any range derivable therein. In one embodiment, the amount of excipient in the pharmaceutical composition is 2% to 5% w / w of the total weight of the pharmaceutical composition.

[0038] II. Manufacturing method Thus, in one aspect, the present disclosure provides pharmaceutical compositions that can be prepared using thin film freezing methods. Such methods are described in US Patent Application Publication No. 2010 / 0221343 and Watts et al., 2013, both of which are incorporated herein by reference. In some embodiments, these methods include dissolving the components of the pharmaceutical composition in a solvent to form a precursor solution. The solvent can be either 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, diethyl ether, ethyl acetate, isopropyl acetate, butyl acetate, propyl acetate, toluene, hexane, heptane, pentane, or combinations thereof. In some embodiments, the precursor solution can include less than 10% w / v of therapeutic agent and excipients. The precursor solution may comprise less than 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% w / v, or any range derivable therein.

[0039] The precursor solution may be deposited on a surface at a temperature that will freeze the precursor solution. In some embodiments, the temperature may be below the freezing point of the solution at atmospheric pressure. In other embodiments, a vacuum may be applied to the surface to freeze the solution below the freezing point at atmospheric pressure. The surface may also be rotated or moved on a moving conveyor-type system, which may distribute the precursor solution evenly on the surface. Alternatively, the precursor solution may be applied to the surface to produce a uniform surface.

[0040] After the precursor solution is applied to the surface, the solvent can be removed to obtain the pharmaceutical composition. Any suitable solvent removal method can be applied, such as evaporation under reduced pressure or elevated temperature, or freeze-drying. In some embodiments, freeze-drying can include reduced pressure and / or low temperature. Such low 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. This temperature is from 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 derivable therefrom. Additionally, the solvent may be removed under reduced pressure of less than 500mTorr, 450mTorr, 400mTorr, 375mTorr, 350mTorr, 325mTorr, 300mTorr, 275mTorr, 250mTorr, 225mTorr, 200mTorr, 175mTorr, 150mTorr, 125mTorr, 100mTorr, 75mTorr, 50mTorr, or 25mTorr.

[0041] Compositions prepared using these methods and the like may exhibit brittleness such that the composition is easily sheared into smaller particles when processed by the device. These compositions not only exhibit high surface area, but also improved flowability of the composition. Such flowability may be measured, for example, by Carr index or other similar measurement methods. In particular, Carr's index may be measured by comparing the bulk density of the powder with the tapped density of the powder. Such compounds may exhibit good Carr index, which may result in better shearing of the particles to produce smaller particles when the composition is processed through a secondary device to deliver the drug.

[0042] III. Definition When used in conjunction with the term "comprising" in the claims and / or specification, the use of the words "a" or "an" may mean "one," but is also consistent with the meanings of "one or more," "at least one," and "one or more." As used herein, "another" may mean at least a second or more.

[0043] As used herein, the terms "drug," "pharmaceutical," "therapeutic agent," and "therapeutically active agent" are used interchangeably to refer to compounds that produce 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 undergone and have received regulatory approval for administration to an organism.

[0044] Use of the term "or" in the claims is used to mean "and / or" unless expressly indicated to refer to alternatives only or unless the alternatives are mutually exclusive. As used herein, "another" may mean at least a second or more.

[0045] As used in the specification and claims, the words "comprising" (and all forms of "comprising", e.g., "comprise" and "comprises"), "having" (and all forms of "having", e.g., "have" and "has"), "including" (and all forms of "including", e.g., "includes" and "include"), or "containing" (and all forms of "containing", e.g., "contains" and "contain") are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0046] As used herein, the term "significant" (and all forms of "significant", e.g., "significantly") is not intended to imply a statistical difference between two values, but merely to suggest the importance of a parameter or the range of differences.

[0047] Throughout this application, the term "about" is used to indicate that values ​​measured using the present methods include the inherent variability of equipment error or the variability that exists between study subjects or experimental studies. Unless another definition is applicable, the term "about" refers to ±10% of the indicated value.

[0048] As used herein, the term "substantially free of" or "substantially free" in reference to a particular component is used herein to mean that the particular component is not intentionally incorporated into the composition at all and / or is present only as a contaminant or in trace amounts. The total amount of all entrapment, by-products, and other materials is present in the composition in an amount of less than 2%. The term "more substantially free of" or "more substantially free" is used to indicate that the composition contains less than 1% of a particular component. The term "essentially free of" or "essentially free" means that the particular component contains less than 0.5%.

[0049] As used herein, the term "domain" refers to a specific region of a composition that is substantially composed of a single material that has properties that are distinct from other components of the composition. "Discrete domain" refers to an individual region of a composition that is a distinct and separate region of the composition. A domain may be substantially composed of a single element from the composition. These domains may be discontinuous such that the separate domains exist as multiple domains that do not contact each other.

[0050] As used herein, the term "nanoparticle" has its customary and usual definition and refers to a discrete particle that behaves as a whole unit, rather than as individual molecules within a particle. Nanoparticles can have a particle size of about 1 to about 10,000 nm, while ultrafine nanoparticles have a particle size of 1 nm to 100 nm, fine particles have a particle size of 100 nm to 2,500 nm, and coarse particles have a particle size of 2,500 nm to 10,000 nm. In some embodiments, the nanoaggregates described herein may comprise a composition of multiple nanoparticles and may have a size of about 10 nm to about 100 μm.

[0051] Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as possible, however, any numerical values ​​inherently contain certain errors necessarily resulting from the standard deviation found in their respective testing measurements and parameters.

[0052] Other objects, features, and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that this detailed description and the specific examples, while indicating preferred embodiments of the present disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. EXAMPLES

[0053] IV. Working Examples In order to facilitate a better understanding of the present disclosure, the following examples of specific embodiments are given. Those skilled in the art will understand that the techniques disclosed in the following examples represent techniques that the inventors have found to work well in implementing the present disclosure, and therefore can be considered to be preferred modes for its implementation. However, those skilled in the art will understand in light of the present disclosure that many changes can be made in the specific embodiments disclosed and still obtain the same or similar results without departing from the spirit and scope of the present disclosure. The following examples should in no way be interpreted as limiting or defining the entire scope of the present disclosure.

[0054] [Example 1] Observations and Results A. Physicochemical properties of voriconazole dry powder formulations Crystalline voriconazole powder formulations containing mannitol were produced using the TFF technique (see Table 1). XRPD and mDSC were primarily used to measure the crystallinity of the formulations. The TFF-VCZ powder formulations containing mannitol were identified as crystalline as shown in Figures 1 and 2. The TFF-VCZ-MAN powder formulations exhibited characteristic voriconazole peaks in XRPD corresponding to voriconazole bulk powder (e.g., 12.4° 2θ and 13.6° 2θ) as well as δ-mannitol (e.g., 9.5° 2θ and 20.2° 2θ), as shown in Figure 1. These indicate that the powder formulations consist of crystalline voriconazole and δ-mannitol. The intensity of the δ-mannitol peaks decreased as the amount of mannitol (% w / w) in the TFF-VCZ-MAN powder formulations decreased, and the peaks corresponding to δ-mannitol were undetectable when the powder formulation contained 5% (w / w) mannitol. The TFF-MAN dry powder was predominantly the δ form, with trace amounts of α and β forms detected by XRPD (13.5° 2θ and 14.5° 2θ, respectively).

[0055] Table 1. Summary of voriconazole dry powder formulations investigated using thin film freezing (TFF) technique. TIFF2024170583000002.tif95144

[0056] The crystallinity of the TFF-VCZ-MAN powder formulations was also confirmed by mDSC. Figure 2 shows that no glass transition was 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 were detected. TFF-VCZ had a melting endothermic peak at 130.86°C with a heat of fusion of 105.3 J / g. The expected heat of fusion of voriconazole in TFF-VCZ-MAN powder was calculated as a percentage (w / w), and the heat of fusion of voriconazole in TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50 was 100.0 J / g and 52.6 J / g, respectively. The measured heats of fusion of voriconazole were 95.1 J / g for TFF-VCZ-MAN 95:5 and 33.7 J / g for TFF-VCZ-MAN 50:50, which were 95.1% and 64.0% of the expected values. TFF-MAN had a melting endothermic peak at 167.31° C. with a heat of fusion of 187.5 J / g. The expected heats of fusion of mannitol in TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50 were 9.38 J / g and 93.8 J / g, respectively. The measured heats of fusion of mannitol were 2.63 J / g and 63.2 J / g, which were 28.0% and 67.4% of the expected values, respectively. Table 2 shows the composition ratios of voriconazole to mannitol (voriconazole:mannitol w / w) in the two formulations tested by mDSC. These ratios are: 1 The ratio was calculated by integration of the proton peaks using H-NMR. The theoretical ratio of one proton for TFF-VCZ-MAN 95:5 was 1:0.1009 and the experimental ratio was calculated to be 1:0.0992, indicating that 98.3% of the expected amount of mannitol was found in TFF-VCZ-MAN 95:5. For TFF-VCZ-MAN 50:50, 100% of the expected amount of mannitol was found. 1 Detected by H-NMR.

[0057] (Table 2) 1Quantitative comparison of voriconazole and mannitol in TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50 by H-NMR TIFF2024170583000003.tif46158

[0058] The particle morphology of the TFF-VCZ-MAN powders is shown in Figure 3. Aggregation of micron-sized particles was observed in the TFF-VCZ powder, and these particles were also seen in other TFF-VCZ-MAN powder formulations. A more porous matrix was observed in the TFF-VCZ-MAN powders containing higher amounts of mannitol. The 3D topography and illustration of the TFF-VCZ and TFF-VCZ-MAN 95:5 powders shown in Figure 4 confirm that the surface texture of the TFF-VCZ-MAN 95:5 powder is rough, whereas the surface texture of the TFF-VCZ powder is smooth. The high-resolution topography of the TFF-VCZ-MAN 95:5 powder in Figure 5 shows that the TFF-VCZ-MAN 95:5 powder is a nanoaggregate consisting of nanoparticles of about 150-500 nm. The SSA of these TFF-VCZ-MAN powders is shown in Figure 6. The TFF-VCZ powder has the lowest SSA (8.36 m). 2 / g, and the porous matrix of the TFF-MAN dry powder showed the highest SSA (17.11 m 2 The SSA showed a 100% increase in the mannitol content (1.0 g / g) of the porous matrix. As more mannitol was added to the TFF-VCZ-MAN powder formulation, the SSA increased. Nitrogen, oxygen, and fluorine were detected by SEM / EDX as shown in Figure 7, and the micron-sized particles were identified as consisting of voriconazole nanoaggregates. Nitrogen and fluorine were not detected, but oxygen was detected, identifying the porous matrix as mannitol.

[0059] The FT-IR peak pattern of the TFF-VCZ powder was consistent with that of the voriconazole bulk powder, and this same peak pattern was also observed in the TFF-VCZ-MAN powders containing various amounts of mannitol. The peak pattern of TFF-MAN was also observed in the TFF-VCZ-MAN powder. Thus, as shown in Figure 8, peaks corresponding only to TFF-VCZ and TFF-MAN were observed in the TFF-VCZ-MAN powder, and no new peaks were observed in the FT-IR spectrum of the TFF-VCZ-MAN powder. 1D ssNMR 13 C and 19 The FCP-MAS spectrum is shown in Figure 9. Voriconazole was synthesized by the reaction with mannitol. 13 There is no spectral overlap of the C peaks, and all resonances are 19 Voriconazole shows identical spectra in TFF-VCZ and TFF-VCZ-MAN. Furthermore, the sharp peak in the spectrum of TFF-VCZ-MAN 90:10 13 C and 19 The F peak confirms the crystallinity of both voriconazole and mannitol. 1 H- 13 The C HETCOR spectrum was compared with that of TFF-VCZ. No intermolecular cross peaks between voriconazole and mannitol were observed in TFF-VCZ-MAN 90:10.

[0060] B. In Vitro Aerosol Performance and Stability The aerodynamic particle size distribution of the TFF-VCZ-MAN powder formulations was measured by NGI and the FPF (% of metered dose) is shown in Figure 11. Based on the FPF (% of metered dose) data, the TFF-VCZ-MAN powder formulation consisting of 90-97% (w / w) voriconazole showed the greatest aerosolization. The FPF (% of metered dose) of TFF-VCZ-MAN 97:3 was significantly higher (p<0.05) than that of TFF-VCZ, with a 66% improvement in FPF (% of metered dose). There was no significant difference in the aerosol performance of TFF-VCZ-MAN powders containing 90-97% (w / w) voriconazole (p>0.05). The aerosol performance of the TFF-VCZ-MAN powder formulations was reduced when more than 10% (w / w) mannitol was included in the composition.

[0061] The effect of physical force on the aerosol performance of TFF-VCZ-MAN 95:5 powder formulation was also investigated by measuring the FPF using NGI. Changes in particle size distribution and aerosol performance with various shear force times were observed as shown in Figure 12 and Figure 13. At 15, 30, and 60 minutes, the FPF (% of metered value) was 44.3, 47.5, and 42.4%, respectively, and the FPF (% of delivered dose) was 68.7, 73.6, and 69.5%, respectively. The initial values ​​before the application of shear force were 40.0% for FPF (% of metered dose) and 58.8% for FPF (% of delivered dose). Changes in MMAD were also observed from 3.7 μm at the initial time to 3.2, 3.0, and 3.1 μm at 15, 30, and 60 minutes, respectively, but no significant changes were observed for GSD.

[0062] Stability studies were performed at 25°C / 60% RH to observe the changes in purity and aerosol performance of the TFF-VCZ-MAN 95:5 powder formulation over a 13-month period, as shown in Figure 14. The purity of voriconazole in TFF-VCZ-MAN 95:5 was maintained, and no degradants were detected over the study period. To compare aerosol performance over the stability study, FPF (% of measured value), FPF (% of delivered dose), MMAD, and GSD were compared at each time point. There were no statistically significant differences in FPF (% of measured value) and FPF (% of delivered dose) over the 13-month period (both p>0.05). GSD after 1 month decreased from the initial value (p<0.05), while MMAD showed no difference over the 13-month period (p>0.05).

[0063] C. Dissolution of Voriconazole Dry Powder Formulations For dissolution testing of TFF-VCZ-MAN powder formulations, PBS at pH 7.4 was used as the receptor medium, and the top of the donor chamber of the Franz cell was covered with parafilm to prevent loss of dissolution medium due to evaporation. The dissolution rate of crystalline TFF-VCZ-MAN 95:5 was compared to amorphous TFF-VCZ-PVPK25 25:75, and the crystalline dry powder showed significantly slower cumulative drug release over the study period (p<0.05), as shown in Figure 15. The cumulative voriconazole release at 3 hours for amorphous TFF-VCZ-PVPK25 was 63.2%, while the cumulative voriconazole release at 3 hours for crystalline TFF-VCZ-MAN 95:5 was only 22.8%. The cumulative voriconazole released at 3 hours for TFF-VCZ-MAN 25:75 and TFF-VCZ-MAN 50:50 was 46.3% and 35.3%, respectively.

[0064] D. Characterization of Voriconazole Dry Powder Formulations Voriconazole (Beinborn et al., 2012b; Ramos and Diogo 2016) and mannitol (Yu et al., 1998) have a strong tendency to crystallize and have glass transition temperatures below room temperature. Therefore, TFF-VCZ-MAN was hypothesized to be crystalline unless there is a strong intermolecular interaction between voriconazole and mannitol to prevent crystallization. The TFF-VCZ-MAN powder formulation was crystalline based on the XRPD data and sharp 1D CP-MAS spectrum, indicating that there is not a strong enough interaction between voriconazole and mannitol.

[0065] Although XRPD is useful for characterizing the crystallinity of powders, it may fail to detect small amounts of amorphous components in the formulation. Therefore, mDSC was performed on the TFF-VCZ-MAN powder, which showed that the TFF-VCZ-MAN dried powder was crystalline. This was because only two endothermic melting peaks were detected for voriconazole and mannitol. However, a melting point depression of mannitol was observed, especially in TFF-VCZ-MAN 95:5. The lower heat of fusion of mannitol in TFF-VCZ-MAN 95:5 may be due to a relatively small amount of mannitol dissolving in the molten voriconazole before the temperature reached the melting point of mannitol. Also, the mannitol particles in TFF-VCZ-MAN 95:5 are usually 100-200 nm, and these nanoscale particles may reduce the heat of fusion. To confirm the efficacy of mannitol in the TFF-VCZ-MAN powder that showed a melting point depression, the molecular ratio of voriconazole to mannitol was adjusted to 100-200 nm. 1 The voriconazole and mannitol ratios were measured by H-NMR. Although NMR is commonly used for qualitative analysis, quantitative NMR analysis is also applicable (Espina et al., 2009; Pauli et al., 2012). The experimental molar ratios of voriconazole to mannitol were in good agreement with the theoretical values ​​for both TFF-VCZ-MAN 95:5 and TFF-VCZ-MAN 50:50. Furthermore, 13 C and 19F ssNMR has often been used to confirm crystalline polymorphism and identify low levels of amorphous drug in solid dosage forms (Correa-Soto et al., 2017; Offerdahl et al., 2005). 13 C and 19 Identical peak positions and linewidths of voriconazole resonances in the F CP-MAS spectrum confirm crystallinity and suggest the absence of a quantifiable amorphous component.

[0066] FTIR was used to study the chemical interactions between voriconazole and mannitol. The hydroxyl groups of voriconazole are involved in its degradation pathway (Shaikh and Patil 2012) and may be the most active sites if there is an intermolecular interaction. If so, this would result in a 3100 cm -1 ~3500cm -1 The FT-IR peaks of voriconazole in the range of 3118.9 cm shift (Silverstein et al., 2005). There are two peaks in this range corresponding to voriconazole, and these peaks are at 3118.9 cm. -1 and 3198.4 cm -1 These two peaks were observed in all of the TFF-VCZ-MAN and TFF-VCZ powder formulations, and no shifts in these peaks were found. For mannitol in this range, the peaks at 3276.6 cm -1 When there was an interaction between mannitol and the aromatic secondary amine of voriconazole, a peak was observed at 1230 cm -1 ~1300cm -1 A peak shift may be observed between 1241.5 cm and 1242.5 cm in this range (Silverstein et al., 2005). -1 , 1248.8cm -1 , 1268.5cm -1 , and 1277.6 cm -1Four peaks from voriconazole were detected in the FT-IR spectra, but no significant peak shift was observed when mannitol was included in the voriconazole powder formulation. Therefore, these FT-IR data support the absence or very weak interaction between voriconazole and mannitol in the TFF-VCZ-MAN powder formulation.

[0067] FT-IR is usually used to identify conformation and intermolecular interactions, while ssNMR can provide more detailed atomic-level information for structural investigation (Tian et al., 2017). In this study, 1D 13 C and 19 Conformational changes were investigated using F CP-MAS. All voriconazole peaks showed no difference in chemical shifts between TFF-VCZ and TFF-VCZ-MAN 90:10. Furthermore, 2D NMR spectroscopy was used to investigate the structural perturbations with better resolution. 13 C- 1 The H HETCOR spectrum was obtained. The results are shown in the direct dimension. 13 This confirms the absence of any change in the chemical shift of C. At the given resolution, the chemical shifts of all aliphatic and aromatic protons in the indirect dimension also show no observable change. Furthermore, the 2D 13 C- 1 H HETCOR has been utilized to detect drug-excipient interactions. At a given spectral intensity, no intermolecular cross-peaks, i.e., no interactions, were observed between voriconazole and mannitol.

[0068] In the TFF-VCZ-MAN powder formulation, two different shaped particles were observed, and based on the observed particle morphology of TFF-VCZ and TFF-MAN, 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 by SEM / EDX. However, the locations where oxygen, fluorine, and nitrogen were detected overlapped each other during the first SEM / EDX run, which indicated particles composed of both voriconazole and mannitol. The cause of this was identified later. Since the measurement depth of EDX is on the micron scale, the detection beam passed through the entire depth of the particles of the tested TFF-VCZ-MAN 50:50 powder. To overcome this issue, the powder was widely dispersed on carbon tape on the sample holder and spot analysis was performed to determine the chemical composition of the two different morphologies of the particles. Spot analysis identified the micron-sized particles as voriconazole nanoaggregates based on their oxygen, nitrogen, and fluorine chemical composition, while the porous matrix was identified as mannitol, which showed an oxygen chemical composition without nitrogen and fluorine. It was therefore concluded that crystalline mannitol phase separated from crystalline voriconazole during the TFF process.

[0069] The AFM images in Figure 5 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 about twice that of TFF-VCZ. This may be because the voriconazole particles are nanoaggregates with a larger specific surface area than is visually seen in the SEM images.

[0070] E. Mannitol level affects aerosol performance and dissolution rate The amount of mannitol in the TFF-VCZ-MAN powder influenced the powder morphology. When a small amount of mannitol was included, submicron mannitol particles were formed as a result of the high degree of supercooling during the TFF process by preventing particle growth (Engstrom et al. 2008). These particles were present on the surface of the voriconazole nanoaggregates and modified the surface properties of the nanoaggregates. These submicron mannitol particles were not removed from the surface of the voriconazole nanoaggregates during aerosolization. This could be because it was difficult to remove nano-sized particles from the surface. Cohesion and adhesion forces are proportional to the particle diameter, whereas removal forces are proportional to the cube of the diameter for gravity, vibration, and centrifugal forces (Hinds 1999). Therefore, it was difficult to separate the submicron mannitol particles from the voriconazole nanoaggregates and the rough surface properties of the voriconazole nanoaggregates were maintained during aerosolization, resulting in a higher degree of aerosolization. Increasing the amount of mannitol in the TFF-VCZ-MAN powder resulted in large porous mannitol matrices. These matrices were not only present on the surface of the voriconazole nanoagglomerates but also surrounded the particles. Multiple voriconazole nanoagglomerates were aggregated because the large porous mannitol matrices maintained the aggregated state of multiple voriconazole nanoagglomerates. These aggregate structures were maintained during aerosolization. As a result, these large aggregate particles reduced the aerosol performance of TFF-VCZ-MAN powder formulations containing more than 10% (w / w) mannitol.

[0071] The aerosol performance of DPI formulations is highly dependent on particle cohesion and adhesion forces. These forces include van der Waals forces, surface tension of adsorbed liquid film, and electrostatic forces (Hickey et al. 1994). All of these are influenced by particle shape and size, surface roughness / texture, relative humidity, temperature, duration and speed of particle contact (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 dipoles between molecules, van der Waals forces decrease significantly when the distance between the surfaces of particles reaches the separation distance (Hinds 1999). Thus, the rougher the surface, the more average particle spacing is maintained, and thus the van der Waals forces decrease crucially. Surface roughness affects not only van der Waals forces but also the surface tension caused by surface moisture. Smooth particle surfaces and high relative humidity lead to stronger surface tension forces. However, electrostatic forces are particle size dependent. Particles larger than 0.1 μm can generate electrostatic forces (Hinds 1999). This attractive electrostatic force is stronger for larger particles and is also related to the relative humidity; that is, lower humidity allows particles to retain their charge for longer. Nevertheless, electrostatic forces are usually considered to be smaller than van der Waals forces and surface tension forces (Hinds 1999). Thus, particle surface roughness and texture play an important role in the aerosol performance of DPI formulations.

[0072] The morphological changes in the powder formulation caused by various amounts of mannitol significantly affected the aerosol performance of the TFF-VCZ-MAN powder formulation. Aerosol performance is altered by changes in particle cohesion and adhesion forces, and the decrease in these forces is related to a decrease in the contact area between particles (Beach et al. 2002) in addition to an increase in particle spacing (Hinds 1999). As shown in Figure 4, the inclusion of small amounts of submicron mannitol particles significantly decreased the contact area of ​​TFF-VCZ-MAN nanoaggregates, leading to further spacing of voriconazole particles. Compared to TFF-VCZ powder, TFF-VCZ-MAN 99:1 powder showed a significant improvement in FPF (% of metered dose) (p<0.05). This improvement with the addition of mannitol continued until 3% (w / w) mannitol was added to the formulation. By adding 1% (w / w) mannitol to formulations containing 97%–100% (w / w) voriconazole, an approximately 5% increase in FPF (% of metered dose) was achieved. Furthermore, the TFF-VCZ-MAN 95:5 powder showed an approximately 30% higher emitted dose compared to the TFF-VCZ powder (68% vs. 36%, respectively, data not shown). This increase in emitted dose was achieved as a result of reduced adhesion of the particles to the device. TFF produces TFF-VCZ-MAN powders that contain very small amounts of moisture (less than 0.1% w / w, data not shown), and because voriconazole and mannitol are not hygroscopic, the surface tension on these particles is expected to be relatively low. Storing the powder in a low humidity environment can generate electrostatic forces, although electrostatic forces are thought to be much smaller than van der Waals forces and surface tension (Hinds 1999). Therefore, decreasing the particle contact area and spacing by modifying the surface properties was mainly responsible for decreasing the cohesive and adhesive forces of the TFF-VCZ-MAN powder formulations, which led to improved aerosol performance. Young et al. also described a relationship between aerosol performance and the separation energy between particles (Young et al. 2002), which is in good agreement with our results.

[0073] Varying amounts of mannitol in the TFF-VCZ-MAN powders affected the dissolution rate as well as the aerosol performance. TFF-VCZ-MAN powders containing higher amounts of mannitol showed increased dissolution rates, which could be explained by faster wetting of the powder by mannitol. In the case of TFF-VCZ-MAN powders containing higher amounts of mannitol, the surrounding mannitol particles surrounding the voriconazole were wetted and dissolved very quickly. Thus, the voriconazole nanoaggregates were surrounded by the dissolution medium in a short time, resulting in a faster dissolution rate. SEM images of TFF-VCZ-MAN 25:75 powder showed that most of the mannitol particles were dissolved in less than 5 minutes on the Franz cell, while submicron mannitol particles were still observed on the surface of the voriconazole nanoaggregates from TFF-VCZ-MAN 95:5 powder. This indicated that the voriconazole nanoaggregates were not wetted as quickly when the powder formulation contained only a small amount of mannitol.

[0074] F. Advantages of the TFF process Highly potent nanoaggregates of voriconazole powder formulations were produced by TFF. Although carrier-free DPI formulations have been reported previously (Yazdi and Smyth 2016a; Yazdi and Smyth 2016b), carriers are commonly included in DPI formulations. However, carrier-based DPI formulations generally have low efficacy. In addition, 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 affect the aerosol performance of the powder during aerosolization, and these factors negatively affect the uniformity of the deposited dose (Du et al. 2017). By using TFF, maximum aerosol performance of TFF-VCZ-MAN nanoaggregates was achieved with only 3% (w / w) mannitol. Thus, the potency of the optimized TFF-VCZ-MAN powder formulation can be up to 97% (w / w). This high potency combined with very low levels of excipients can result in less powder needing to be delivered, eliminating problems commonly caused by carriers, such as low potency and non-uniform deposition.

[0075] High-potency DPI formulations can also be produced by other techniques, such as, for example, milling. Although the size of particles produced by milling and suitable for pulmonary delivery is a few microns, such particles are considered to be single, discrete, micron-sized particles. Voriconazole DPI formulations produced by TFF, which are nanoaggregates, may have significantly higher total pulmonary absorption efficiency and uniformity of dose distribution based on a study by (Longest and Hindle 2017). These voriconazole nanoaggregates are expected to allow better epithelial coverage when fungal colonies are present. TFF was able to produce nanoaggregates. This is because rapid nucleation at freezing rates of up to 10,000 K / s allowed for narrower particle size distribution and lower Ostwald ripening, forming more nuclei during the freezing process and preventing particle growth (Engstrom et al. 2008; Overhoff et al. 2009). The small size of the unfrozen channel and the rapid increase in viscosity of the unfrozen solution (Engstrom et al. 2008) resulted in voriconazole nanoaggregates of similar size.

[0076] Surface modification of particles can also be performed by TFF. Begat et al. previously reported surface modification of particles using hydrophobic materials, e.g., lecithin, leucine, and magnesium stearate. Although particles treated by dry mechanical fusion methods such as mechanofusion showed improved aerosol performance with or without a carrier by lowering the surface free energy (Begat et al. 2005; Begat et al. 2009), this method was based on mixing the drug with force control agents such as lecithin, leucine, and magnesium stearate. The mechanofusion method requires the input of mechanical energy into the formulation, which may cause chemical instability of the drug. Furthermore, surface modification by mixing is only applicable to discrete micron-sized particles and may not be applicable to nanoaggregates, since mixing may disaggregate the aggregates. Kawashima et al. also reported surface modification of particles by various methods, e.g., mechanical shear mixing, freezing, or spray drying (Kawashima et al. 1998). With the use of hydrophilic additives such as light anhydrous silicic acid (AEROSIL), the surface of hydrophobic particles was converted to hydrophilic, and the surface-modified particles showed improved inhalation behavior in vitro. However, this method uses discrete micron-sized drug particles and cannot be used for nanoaggregates. Therefore, these discrete micron-sized particles treated with other methods cannot achieve the increased uptake and microdosimetry of nanoaggregates as described in Longest and Hindle 2017. However, in TFF, no energy input was required to modify the surface of the particles. Surface modification of voriconazole nanoaggregates with phase-separated submicron mannitol particles, which were individually present on the surface of the drug nanoaggregates, was performed by a fast freezing rate that prevented particle growth.

[0077] Highly potent (up to 97% w / w) nanoaggregates of crystalline voriconazole powder formulations intended for dry powder inhalation were successfully developed using TFF technology. Small amounts of mannitol used as the sole excipient advantageously enhanced the aerosol performance of voriconazole nanoaggregates with phase-separated submicron crystalline mannitol acting as a surface texture modifier. Voriconazole dry powder for inhalation produced by TFF is a viable topical treatment option for invasive pulmonary aspergillosis with high aerosolization efficiency and drug loading while offering potential advantages regarding nanoaggregate deposition in the airways.

[0078] Example 2 Materials and general methods A. Material The following materials were purchased: voriconazole (Carbosynth, San Diego, CA); Kollidon® 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) was used, and Pearlitol® PF, pyrogen-free mannitol, was donated by Roquette America Inc. (Geneva, IL).

[0079] B. Preparation of Powders for Dry Powder Inhalation Using TFF Mannitol and voriconazole (30–100% w / w) powders were dissolved in a mixture of acetonitrile and water (50:50 v / v) to maintain the solids in the solution at 1% w / v. Approximately 15 μL of each solution was dropped from a height of 10 cm onto a rotating cryogenically cooled (−60°C) stainless steel drum. Frozen samples were collected in a stainless steel container filled with liquid nitrogen and transferred to a −80°C freezer until transfer to the freeze dryer. Solvent was removed using a VirTis Advantage Lyophilizer (VirTis Company Inc., Gardiner, NY). Samples were kept at −40°C for 21 h, the temperature was slowly increased to 25°C over 21 h, and then kept at 25°C for an additional 21 h to dry. Pressure was maintained at 100 mTorr during the drying process.

[0080] CX-ray powder diffraction (XRPD) The crystallinity of the powder samples was measured by X-ray diffraction (MiniFlex 600, Rigaku Corporation, Tokyo, Japan) measuring from 5 to 35° 2θ (0.02° step, 3° / min, 40 kV, 15 mA).

[0081] D. Scanning Electron Microscopy (SEM) The surface morphology of the samples was identified using a SEM (Zeiss Supra 40V SEM, Carl Zeiss, Heidenheim an der Brenz, Germany). An aliquot of the powder was placed on carbon tape and sputter-coated with 60 / 40 Pd / Au for 20 min before imaging.

[0082] E. Modulated Differential Scanning Calorimetry (mDSC) Thermal analysis of the powder samples was studied by differential scanning calorimetry model Q20 (TA Instruments, New Castle, DE) equipped with a refrigerated cooling system (RCS40, TA Instruments, New Castle, DE). Modulated DSC was performed with a modulation period of 50 s, a modulation amplitude of 1 °C, and an average heating rate of 5 °C / min. Tzero pans and Tzero airtight lids from TA Instruments were used to hold the samples during testing, and the lids were punctured with a 20 G syringe needle before placing the pans in the sample holder.

[0083] F. Scanning Electron Microscopy-Energy Dispersive X-ray Spectroscopy (SEM / EDX) SEM / EDX (Hitachi S5500 SEM / STEM, Hitachi America, Tarrytown, NY) was used to identify the elements of the powders produced by TFF.

[0084] G. Atomic Force Microscopy (AFM) Two different atomic force microscopy techniques were used during this study. Three-dimensional (3D) surface topography images of particles generated by TFF were acquired by an Asylum MFP-3D AFM (Oxford Instruments, Oxfordshire, UK) equipped with an aluminum-coated MikroMasch HQ:NSC15 cantilever (NanoWorld AG, Neuchâtel, Switzerland) with a resonant frequency of 325 kHz, a force constant of 40 N / m, and a typical tip radius of 8 nm. The powder was affixed to the AFM disk with carbon tape, and particles not firmly attached to the carbon tape were blown off using compressed nitrogen gas. Topography was performed in tapping mode with a scan speed of 1.00 Hz, a set point of 1.08 V, and an integral gain of 20.0. Feedback filters, drive amplitudes, and drive frequencies were optimized for each sample, and all images were collected at a resolution of 512 × 512. 3D topographic images were generated using Gwyddion software (Necas and Klapetek 2012) (64-bit Windows version 2.50).

[0085] 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, Neuchâtel, Switzerland) with a resonant frequency of 330 kHz, a force constant of 42 N / m, and a tip radius of less than 7 nm was used. A 380 μm single-sided polished P-type silicon wafer was coated with Tween® 20 (VWR, Radnor, PA) before loading the powder sample for AFM. Tween® 20 (1.5% w / v) was previously dissolved in HPLC-grade methanol (Fisher Scientific, Pittsburgh, PA). The solution was dropped onto the silicon wafer using a transfer pipette, and the solution was removed by compressed nitrogen gas. The powder was placed in a DP4 insufflator (Penn-Century Inc., Windmoor, PA) and aerosolized onto the silicon wafer using a 3 mL syringe. After the aerosolized powder was loaded onto the silicon wafer, compressed nitrogen gas was used to remove the powder solids that were not strongly attached to the silicon wafer. Tapping mode was applied to collect images at a resolution of 512 × 512 with a scanning speed of 0.30 Hz. Other values ​​of the AFM were optimized for each sample. Topography images were processed with Gwyddion software (Necas and Klapetek 2012) (64-bit Windows version 2.50).

[0086] H. Aerodynamic particle size distribution analysis Aerodynamic particle size was measured by a Next Generation Pharmaceutical Impactor (NGI) (MSP Co. Shoreview, MN) connected to a High Capacity Pump (Model HCP5, Copley Scientific, Nottingham, UK) and a Critical Flow Controller (Model TPK 2000, 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 induction port at a flow rate of 60 L / min for 4 seconds per actuation. No pre-separator was used throughout the study. The collection plate of the NGI was coated with a 2% w / v solution of polysorbate 20 in methanol and allowed to dry for 20 minutes before use. After aerosolization, the powder was extracted with a mixture of water and acetonitrile (50:50 v / v) and the voriconazole content was analyzed by HPLC. Mass median aerodynamic diameter (MMAD), geometric standard deviation (GSD), and fine particle fraction (FPF) were calculated based on the dose deposited on the device, induction port, stages 1-7, and micro-orifice collector (MOC) using Copley Inhaler Testing Data Analysis Software (CITAS) version 3.10 (Copley Scientific, Nottingham, UK).

[0087] I. High Performance Liquid Chromatography (HPLC) 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 method details are as follows: an isocratic method for aerodynamic properties, using a mobile phase of 40 / 60 (v / v) water / acetonitrile with 0.1% (v / v) TFA and a flow rate of 0.8 mL / min for 4 min, and a gradient method for chemical degradants during stability testing. In the gradient method, acetonitrile with 0.1% (v / v) TFA was gradually increased from 25 to 95% (v / v) over 14 min, mixed with water with 0.1% (v / v) TFA, and a flow rate of 0.8 mL / min. In both methods, samples were analyzed at a detection wavelength of 254 nm at 25° C. Linearity was performed from 50 ng / mL to 100 μg / mL using an injection volume of 15 μL.

[0088] J. Solution Nuclear Magnetic Resonance (Solution NMR) 1 H NMR was performed to calculate the weight ratio of voriconazole to mannitol in the TFF-VCZ-MAN powder. 1 H NMR spectra were obtained using a VNMR 600 (Varian, Palo Alto, CA) spectrometer at 25 °C using dimethyl sulfoxide- d6 (DMSO- d6 ) at 600 MHz. Chemical shifts are recorded relative to DMSO-d6 at 2.47 ppm.

[0089] K. Solid state nuclear magnetic resonance (ssNMR) ssNMR experiments were performed on a Bruker Avance III HD 400 MHz spectrometer (Bruker, Billerica, MA) at 25 °C and a magic angle spinning (MAS) frequency of 12 kHz. A Bruker 4 mm triple resonance HFX probe was used. 1 H / 13 C or 1 H / 19All samples were run in double resonance mode tuned to the F frequency. 2 The rotor (Wilmad-LabGlass, PA) was filled with 1D 13 C and 19 F cross-polarization (CP) MAS experiments were performed using 1 A linear gradient power level of 80–100 kHz was performed during a contact time of 2 ms in the H channel. High-power SPINAL 64 proton decoupling was used at a field strength of 80 kHz. The same power parameters, contact times, and MAS frequencies were used for two-dimensional (2D) 13 C- 1 Adamantine was used in H CP heteronuclear correlation (HETCOR) experiments. 13 Ethyl C was used as an external standard to calibrate the C chemical shifts. 13 The C peak was referenced at 38.48 ppm.

[0090] L. Fourier transform infrared spectroscopy (FT-IR) A Nicolet™ iS™ 50 FT-IR equipped with a Smart OMNI-Sampler™ (ThermoFisher Scientific, Waltham, MA) was used to study the intermolecular activity between voriconazole and mannitol in TFF-VCZ-MAN powder. Measurements were performed using the samples as dry powders and were measured at 4000–700 cm. -1 A spectral range of was recorded with an aperture of 150, a resolution of 4 and 32 scans.

[0091] M.Brunauer-Emmett-Teller (BET) specific surface area (SSA) analysis The SSA of TFF-VCZ-MAN powders was measured by one-point BET method using a Monosorb™ Rapid Surface Area Analyzer Model MS-21 (Quantachrome Instruments, Boynton Beach, FL). To remove surface impurities, samples were degassed with 20 psi nitrogen gas at ambient temperature for 24 h. A mixture of nitrogen / helium (30:70 v / v) was used as the adsorption gas.

[0092] N. Shear resistance test To test the shear resistance of the TFF-VCZ-MAN 95:5 powder, the powder was placed in a stainless steel container (2 7 / 8" inner diameter, 4 1 / 4" height) and pre-sheared by rotating the container at 85 rpm. Powder samples were taken at 15, 30, and 60 minutes and the aerodynamic properties were compared to the initial condition.

[0093] O. Dissolution Test An in vitro dissolution method was used to quantify the dissolution of voriconazole from powders processed by the TFF technique. A Franz cell apparatus was used to allow differentiation of voriconazole release from powders produced by TFF. A Next Generation Pharmaceutical Impactor (NGI) (MSP Co. Shoreview, MN) connected to a High Capacity Pump (Model HCP5, Copley Scientific, Nottingham, UK) and a Critical Flow Controller (Model TPK 2000, Copley Scientific, Nottingham, UK) was used to load aerosolized powder onto a Tuffryn membrane filter (25 mm, 0.45 μm, Pall Corporation, Port Washington, NY). Five nozzles in stage 2 of the NGI lid were blocked with lab tape, leaving only one nozzle open. A Tuffryn membrane filter was placed in the collection cup under the open nozzle in stage 2 and secured in place with lab tape. A #3 HPMC capsule (VCaps plus, Capsugel, Morristown, NJ) containing TFF powder (approximately 5–10 mg) was placed into a high-resistance RS01 dry powder inhaler (Plastiape, Osnago, Italy) and dispersed into the NGI through the USP induction port for 4 seconds at a flow rate of 60 L / min per actuation. No pre-separator was used. After aerosolization, the membrane filter loaded with powder (approximately 0.5–1 mg) was carefully removed from the collection cup and placed on top of the receptor chamber of a Franz cell pre-filled with degassed 10 mM phosphate-buffered saline (PBS), pH 7.4 (5 mL). The donor chamber was placed on top of the membrane filter, and the membrane filter was secured between the receptor and donor chambers with a pinch clamp. Parafilm was used to cover the top of the donor chamber. Dissolution studies were performed at 37 °C sink conditions while a magnetic bar was stirring in the receptor chamber. Dissolution medium (150 μL) was removed at time intervals of 0, 20, 40, 60, 120, and 180 min for HPLC analysis without dilution, and was replaced with fresh dissolution medium after each sampling.

[0094] P. Preparation of SEM Samples During Recovery and Dissolution of Aerosolized Particles SEM images of aerosolized TFF-VCZ-MAN powders before and during dissolution testing were recorded using a Fast Screening Impactor (FSI) (Copley Scientific, Nottingham, UK) connected to a High Capacity Pump (Model HCP5, Copley Scientific, Nottingham, UK) and a Critical Flow Controller (Model TPK 2000, 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 onto a glass fiber filter (MSP Co. Shoreview, MN) placed within the FSI to collect particles with an aerodynamic diameter of 5 μm or less. Once collected on the filter, the particles were transferred to carbon tape by tapping the carbon tape, which had previously been applied to the SEM sample, on the filter and SEM images were recorded.

[0095] To record SEM images during dissolution testing, the glass fiber filters loaded with powder from FSI were cut into circles (25 mm diameter). The glass fiber filters were then placed between the donor chamber of a Franz cell and the receptor chamber previously filled with PBS, pH 7.4, at 37 °C. The filters were left on the Franz cell for 5 min and then placed in a -80 °C freezer for 1 h. The solvent was removed using a VirTis Advantage Lyophilizer (VirTis Company Inc., Gardiner, NY) for 5 h at 25 °C. Carbon tape previously attached to the SEM sample was tapped on the glass fiber filter to transfer the TFF-VCZ-MAN powder, and SEM images were recorded as described above.

[0096] Q. Stability testing TFF-VCZ-MAN 95:5 dry powder was presheared in a glass vial as described in the shear resistance test. 7.6 mg to 8.4 mg of the presheared powder was filled into size #3 HPMC capsules (Capsugel, Morristown, NJ). 14 capsules filled with powder were transferred to a scintillation vial and the vial was purged with nitrogen gas for 20 seconds before being closed with a cap. The vial was sealed with aluminum foil (13 x 15 cm) that had previously been purged with nitrogen gas for 30 seconds and the aluminum foil was maintained at 25°C / 60% RH. Purity and aerosol performance were performed at 1, 3, 6, and 13 months.

[0097] R. Statistical analysis Aerodynamic performance and cumulative drug release were compared for statistical analysis by Student's t-test. P values ​​less than 0.05 were considered significant. Data significance was compared using JMP® 10.0.0.

[0098] Example 3: Large-scale manufacturing and inhaler testing of voriconazole compositions 1.Results A. Observation of the cooling process at -60℃ and -150℃ Table 3 shows the various formulations and processing conditions. Figure 16 shows images of the freezing process at two different temperatures. Water / ACN (50:50 v / v) solutions containing voriconazole and mannitol (95:5 w / w) were used at solid loadings of 1% 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 thermal equilibrium was reached within 200 ms. Nucleation was observed at the edge of the sample disk in about 1 / 30 ms, but freezing proceeded from the center of the disk to its edge at -60°C. In contrast, at -150°C, nucleation started within 1 / 60 ms for solutions with solid loadings of 1% and 3% (w / v), and cooling proceeded uniformly throughout the sample disk. However, thermal equilibrium was not reached by 200 ms.

[0099] Table 3. List of formulations and processing parameters TIFF2024170583000004.tif94156

[0100] B. Physical properties of voriconazole nanoaggregates produced by TFF Figure 17 shows high resolution topography of voriconazole nanoaggregates processed at two different temperatures. This figure shows that voriconazole nanoaggregates formed at the lower temperature (-150°C) (Formulation #4) consist of smaller nanoparticles. When processed at -150°C, nanoparticles as small as 200 nm were observed, whereas at -60°C nanoparticles of about 500 nm were found (Formulation #2).

[0101] Figure 18 compares the particle morphology of voriconazole nanoaggregates formed using various processing parameters. When water / ACN (30:70 v / v) (Formulation #1) was used as the solvent system, porous structure of mannitol with particle size greater than 20 μm was observed. Voriconazole nanoaggregates produced with other solvent systems showed modification of the surface properties of voriconazole particles by mannitol nanoparticles. Lower processing temperatures resulted in smaller particles within the solid loading range tested (1-3% w / v).

[0102] Figure 19 shows SEM images of aerosolized voriconazole nanoaggregates produced at -60°C and -150°C (formulations #7 and 6, respectively). The figure shows that the nanoaggregates consist of nanoparticles as small as 200 nm. The voriconazole nanoaggregates remained primarily micron-sized nanoaggregates, although irregularly shaped nanoaggregates were observed that did not completely disaggregate after aerosolization. The surface of these nanoparticles remained modified in nature after aerosolization with the DP4 insufflator.

[0103] C. Comparison of physical and aerodynamic properties with scale-up Table 4 shows the aerodynamic properties and moisture content of voriconazole nanoagglomerates produced at small scale (200 mg) and large scale (90 g). Additionally, Figure 20 compares the crystallinity of the various powder formulations. Comparing large scale to small scale, FPF (% of metered dose, 35.6% vs. 37.0%), FPF (% of delivered dose, 49.5% vs. 48.5%), and MMAD (3.69 μm vs. 3.52 μm) were not significantly different (p>0.05) when tested at a flow rate of 60 L / min using the Plastiape® low resistance RS00 device. Also, the moisture content of both batches was less than 0.1% (w / w) by TGA. XRPD spectra of voriconazole nanoagglomerates showed no pattern difference between small scale and large scale.

[0104] Table 4. Comparison of physicochemical and aerodynamic properties by scale TIFF2024170583000005.tif127151

[0105] D. In vitro aerosol performance i. By co-solvent, processing temperature, and solids loading The effect of co-solvent, processing temperature, and solids loading on the aerosol properties of unconditioned voriconazole nanoaggregates was evaluated using NGI. The results are shown in Table 5. Different ratios of water to acetonitrile in the co-solvent changed the aerosol properties of voriconazole nanoaggregates when the solids loading (1%) and processing temperature (-60 °C) were fixed (formulations #1-3). As the percentage of water increased from 30% (v / v) to 50% (v / v) to 70% (v / v), the FPF (% of metered dose) increased from 34.3% to 37.9% to 45.6%. The FPF (% of delivered dose) also increased (53.1%, 61.2%, and 69.9%, respectively), but the MMAD decreased (3.41, 3.31, and 3.09 µm, respectively).

[0106] Table 5. Aerosol properties by solvent system, processing temperature, and solid loading. TIFF2024170583000006.tif23372 Plastiape low resistance RS-00 device at 60 L / min (n=3; mean ± SD) The powder is not conditioned.

[0107] The effect of processing temperature was also examined. When the processing temperature was decreased from -60°C to -150°C (formulations #2 and 4) with fixed solid loading (1%) and co-solvent (water / ACN 50:50 v / v), the FPF (% of metered dose) significantly increased from 37.9% to 46.7% (p<0.05). However, the FPF (% of delivered dose) and MMAD did not change significantly (61.2% vs. 67.5% and 3.31 vs. 3.27 μm, respectively) (p>0.05).

[0108] Solid loading also affects aerosol properties. As shown in Table 5, higher solid loadings result in poorer aerosol properties. As solid loading increased from 1% to 2% to 3% (formulations #4-6), the FPF (% of metered dose) decreased from 46.7% to 41.3% to 37.0% when the powder was not conditioned. The FPF (% of delivered dose) also decreased from 67.5% to 60.9% to 48.5%. There was no significant difference in the MMAD of 1% and 2% (formulations #4 and 5) (3.27 vs. 3.24 μm, p>0.05), but 3% (formulation #6) resulted in a significantly larger MMAD (3.52 μm, p<0.05).

[0109] ii. By device The aerosol performance of voriconazole nanoagglomerates (formulation #6) was evaluated using four different types of Plastiape devices, namely low and high resistance RS00, and low and high resistance RS01. Table 6 shows the evaluation of the effect of various flow rates on the aerosol performance. At flow rates of 90, 60, and 30 L / min, the low resistance RS00 device showed FPFs (% of metered dose) of 48.6%, 45.8%, and 27.0%, respectively, and FPFs (% of delivered dose) of 63.7%, 63.9%, and 48.9%, respectively. The MMAD increased from 3.22 to 3.36 to 4.32 μm as the flow rate decreased from 90 to 60 to 30 L / min. The high resistance RS00 device showed FPFs (% of metered dose) of 34.7% at 60 L / min and 30.7% at 30 L / min. The MMAD of the high resistance RS00 device was 3.76 μm at 60 L / min and 3.83 μm at 30 L / min. The FPF (% of metered dose) of the low resistance RS01 device at flow rates of 90, 60, and 30 L / min was 40.1%, 35.8%, and 27.0%, respectively, with MMADs of 4.28, 4.37, and 5.34 μm, respectively. The high resistance RS01 showed an FPF (% of metered dose) of 31.7% at 60 L / min and 20.2% at 30 L / min, but MMADs of 4.48 and 5.06 μm, respectively. In general, the low resistance devices showed higher aerodynamic performance at the same flow rates, with the RS00 device resulting in better performance compared to the RS01 during in situ aerosol performance testing.

[0110] (Table 6) Aerosol characteristics by device TIFF2024170583000007.tif233120

[0111] iii. Per dose The effect of varying dose loading on aerosol performance was tested using the high resistance RS00 and high resistance RS01 devices. The results are shown in Table 7. When using the high resistance RS00 device at 60 L / min, increasing the loading dose from 10 mg to 15 mg to 20 mg did not significantly (p>0.05) change the aerosol properties of FPF (% of metered dose) (34.7%, 33.8%, and 31.8%, respectively), FPF (% of delivered dose) (55.0%, 55.5%, and 51.5%, respectively), and MMAD (3.76, 3.77, and 3.84 μm, respectively). However, the high resistance RS01 device at 60 L / min showed significant differences (p<0.05) between the 10 mg and 20 mg loading dose in FPF (% of metered dose) (31.7% vs. 25.3%), FPF (% of delivered dose) (48.5% vs. 37.4%), and MMAD (4.48 vs. 5.21 μm).

[0112] Table 7. Aerosol characteristics by dose TIFF2024170583000008.tif23084

[0113] Table 8. Aerosol characteristics by device based on pressure drop TIFF2024170583000009.tif23675·n=3 Loading: 15.0mg / capsule (fixed)

[0114] 2. Consideration A. Processing design domain of voriconazole nanoaggregates produced by TFF Processing parameters within the design domain of the freezing process used in TFF needed to be considered and their impact during development and subsequent scale-up understood, including solvent system, processing temperature, solids loading, and batch size. The processing design domain was determined using the low-resistance RS00 device at a flow rate of 60 L / min because aerosolization with the low-resistance RS00 device was more dependent on the inhalation flow rate and formulation properties. This dependency could differentiate aerosolization from formulations produced by different processing design parameters.

[0115] B. Solvent Systems The physicochemical properties of amorphous solid dispersions of danazol produced using TFF were not affected by the two different solvents used, tert-butanol and acetonitrile (Overhoff et al., 2007), whereas the crystallinity, morphology, and aerosol performance of voriconazole produced using TFF with PVP K12 or PVP K30 differed depending on the solvent composition, including water and 1,4-dioxane (Beinborn et al., 2012). The water and acetonitrile co-solvent system used during this study was employed to develop tacrolimus formulations, as well as voriconazole formulations produced using TFF (Watts et al., 2013; Moon et al., 2019).

[0116] No differences in crystallinity were observed for the different solvent compositions, but differences in morphology were. Furthermore, significant trends in aerodynamic properties were observed as the solvent composition was changed. Without powder conditioning, solvent compositions containing more water showed enhanced aerosolization. This result may be related to two factors: viscosity of the co-solvent system and freezing phase separation.

[0117] In a co-solvent system of water and acetonitrile, more water increases the viscosity (Thompson et al., 2006; Cunningham et al., 1967). High viscosity may hinder the movement of molecules during the freezing process. Thus, the molecules are more uniformly distributed in the frozen state, and the solute concentration in the unfrozen channel may not increase significantly. A lower viscosity of the solvent may allow more movement of molecules during the freezing process, which may lead to molecular aggregation. As a result, the solute concentration in the unfrozen channel increases. Because the voriconazole powder produced by TFF is a crystalline nanoaggregate, a higher solute concentration may produce larger nanoparticles due to the shorter distance between molecules.

[0118] Although TFF involves ultrafast supercooling, the freezing process of the water / ACN solvent system requires up to 200 ms at -60 °C. Therefore, during this 200 ms freezing time, a higher degree of molecular aggregation may occur in the lower viscosity solvent, resulting in poorer aerosol performance. This trend was also observed in a previous study by Beinborn et al. on voriconazole produced using TFF (Beinborn et al., 2012). When crystalline voriconazole powders containing PVP K12 or PVP K30 were produced in a binary solvent system of water and 1,4-dioxane, a higher degree of aerosolization was obtained for the TFF particles produced in 1,4-dioxane / water (20:80 v / v) compared to particles produced in 1,4-dioxane / water (50:50 v / v). The viscosity of 1,4-dioxane is higher than that of water, whereas 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). Thus, the viscosity of the co-solvent system is one of the factors that influence the aerosol performance after lyophilization.

[0119] Prevention of freezing phase separation is the second possibility for enhancing aerosol performance with more water-containing cosolvent systems. Cosolvent systems consisting of water and acetonitrile are well known to phase separate during the freezing process when they contain 35-88% (v / v) acetonitrile (Gu et al., 1994; Zarzycki et al., 2006). Once phase separation occurs below -1.32 °C (Zarzycki et al., 2006), the unfrozen solvent separates into an 88% (v / v) acetonitrile phase and a 65% (v / v) water phase, allowing solutes to migrate to the phase with higher solubilities (Gu et al., 1994).

[0120] This freeze phase separation occurred in formulation #1, which was treated with water / ACN (30:70 v / v) at -60°C. 5% (w / w) mannitol in the voriconazole nanoaggregates acts as a surface modifier (Moon et al., 2019). Thus, mannitol is observed on the surface of the crystalline voriconazole nanoaggregates, which are nanoparticles, as shown in the SEM images of the other formulations in Figure 18. However, porous mannitol particles are observed at around 20 μm, and these particles have the same morphology as TFF-mannitol (Moon et al., 2019). With mannitol particles of this size, which occurred in formulation #1, there is less mannitol available to act on the surface, which reduces the effectiveness of mannitol in surface modification, thereby resulting in poor aerosolization.

[0121] Although supercooling in TFF can minimize phase separation and produce very small ice channels due to ultrafast freezing (Moon et al., 2016), the processing temperature of -60 °C results in low levels of phase separation in water / ACN (30:70 v / v), which promotes aggregation and increased concentration of mannitol during the freezing process time (up to 200 ms). However, water / ACN (70:30 v / v) does not phase separate during freezing (Gu et al., 1994; Zarzycki et al., 2006). Therefore, aggregation and increased concentration caused by freezing phase separation are unlikely to occur.

[0122] C. Processing temperature In addition to the solvent composition, processing temperature also influences the aerosol performance of crystalline voriconazole nanoaggregates produced using TFF. Lower processing temperatures result in higher degrees of supercooling, which results in smaller ice channels and prevents particle growth (Overhoff et al., 2009; Engstrom et al., 2008). The temperature of -150°C in this study showed much faster nucleation due to ultrafast supercooling. This supercooling at -150°C resulted in smaller nanoparticles in the voriconazole nanoaggregates, which consisted of nanoparticles as small as 200 nm, as observed using both AFM and SEM, due to the prevention of particle growth. In contrast, when processed at -60°C, a particle size of approximately 500 nm was observed using AFM. The smaller the voriconazole nanoaggregates are, the more likely they are to disaggregate into smaller particles during inhalation, thereby leading to improved aerosol performance.

[0123] Interestingly, the level of aerosol performance improvement caused by high degree of supercooling at -150° C. is comparable to that caused by the water / ACN (70:30 v / v) co-solvent system at low degree of supercooling at -60° C. The FPF (% of metered dose) and MMAD are not significantly different (p>0.05) under these two processing conditions (formulations #3 and #4).

[0124] D. Solid load Increasing the solids loading is one way to reduce processing time in the manufacture of powder formulations using TFF. However, increasing the solids loading usually compromises aerosol performance. The bulk density of voriconazole nanoagglomerates produced at a solids loading of 1% (w / v) is approximately 30 mg / cm without conditioning or physical shear. 3 Therefore, to accelerate the manufacturing process, the solids loading was increased to 3% (w / v), which is 30 mg / cm 3This corresponds to a bulk density of 10 ...

[0125] Bulk density at 3% (w / v) solids loading resulted in similar bulk density after lyophilization as 1% (w / v), resulting in poor aerosol performance before powder conditioning. However, proper conditioning improved performance comparable to voriconazole nanoagglomerates produced at 1% (w / v) solids loading using optimized processing parameters (Formulation #4). Therefore, aerosolization studies were performed with Formulation #6 with powder conditioning.

[0126] E. Batch Size Until recently, the TFF process was applied by dropwise feeding of the solution using a syringe or a separatory funnel. As a result, the freezing of the solution required more time. This was a major obstacle to the scale-up of the TFF process. To expedite the freezing process, a two-channel peristaltic pump was applied and the TFF process was used to produce amorphous meloxicam (Jermain et al., 2019). However, during this study on voriconazole, the number of channels was increased to eight and the solution feeding rate 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 prevent the frozen sample disks from overlapping each other when the feeding rate was higher.

[0127] Increasing the rotation speed reduced the time that the frozen samples remained on the cryogenic drum. This time was reduced from 4 to 2 seconds before the samples were collected in a tray containing liquid nitrogen. However, due to the ultrafast freezing of TFF, the time required for the freezing process is typically less than a few hundred milliseconds (Overhoff et al., 2007), and we did not expect that increasing the rotation speed of the cryogenic drum would affect the freezing process. For formulation #6 in this study, nucleation occurred in less than 1 / 60th of a second (Figure 18), and the frozen samples reached thermal equilibrium in less than 2 seconds before being collected in the tray.

[0128] 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). Thus, uniform nucleation was observed throughout the frozen samples. Furthermore, the size of the droplets is similar regardless of scale, so the freezing process is independent of the scale and there are no significant differences in the results for the frozen samples. With an overall similar freezing process, the physicochemical and aerodynamic properties at the small scale were comparable to the larger scale.

[0129] After deciding to use a peristaltic pump in the scale-up of the TFF process, the capacity of the lyophilizer was also tested. The data in Table 4 also confirm that the lyophilization of 90 g of voriconazole nanoaggregates using the three-shelf AdVantage Pro lyophilizer was not different from the 200 mg of voriconazole nanoaggregates lyophilized using the one-shelf AdVantage 2.0 lyophilizer. Therefore, the scale-up of the TFF process using a peristaltic pump with a feed solution flow rate of 25 mL / min is suitable when using the three-shelf AdVantage Pro lyophilizer.

[0130] F. Device interactions with voriconazole nanoaggregates using different devices and flow rates During the development of pharmaceuticals delivered by DPIs, device design or selection is as important as formulation development in terms of aerosol performance. The same powder formulation can be aerosolized variably with different DPI devices (Parumasivam et al., 2017). In this study, we tested the commercially available Plastiape RS01 and RS00 devices, which have been applied in many DPI products marketed or in development (Armer et al., 2016; Elkins et al., 2014; Roscigno et al., 2017). The RS01 and RS00 devices employ the same delivery technology; that is, lifting the capsule from the housing and rotating it at high speed (Dry Powder Inhaler RS01: How to Use: Plastiape; [Available at: plastiape.com / en / content / 1635 / dry--powder-inhaler--rs01--how-use). However, the powder in the RS01 device exits the capsule through two holes, while the RS00 device has a longer mouthpiece and releases the powder through eight smaller holes in the capsule. In comparison, the overall aerosol properties of voriconazole nanoagglomerates using the RS00 device are better when compared using the same flow rate with the same type of resistance.

[0131] This improved performance achieved using the RS00 device may be due to the smaller holes created by the perforation system of the RS00 device. The smaller holes may aid in the deagglomeration of the larger voriconazole nanoagglomerates as they leave the capsule, resulting in a smaller MMAD and a higher FPF as a result of these smaller hole sizes. This may be a unique feature of voriconazole nanoagglomerates for DPIs, as they consist of brittle nanoagglomerates. Other powder formulations for DPIs produced using spray drying or milling may not be considered to be nanoagglomerates. Therefore, the size of the holes as the powder leaves the capsule may not have a significant effect on the overall performance.

[0132] Comparing the low and high resistance of the RS01 and RS00 devices, both low resistance devices generally performed better than the high resistance devices at flow rates of 60 L / min and 30 L / min. The low resistance devices also showed higher ED compared to the high resistance devices. However, deagglomeration and fine dispersion of powders using the low resistance devices is dependent on the patient's inhalation flow rate (Dal Negro RW, 2015), and different inhalation flow rates result in variability in aerosolization. This was also observed with both low resistance devices in this study. A flow rate of 90 L / min achieved maximum aerosolization with the low resistance RS00 device, but a significant decrease in FPF (% of metered dose) was observed at 30 L / min (18.8%). A similar trend was observed when using the low resistance RS01 device. The decrease in FPF (% of metered dose) at flow rates from 60 L / min to 30 L / min was also significant (8.8%) when using the low resistance RS01. However, with the high resistance RS00 device, the difference in FPF (% of metered dose) was only 4.0% between flow rates of 60 and 30 L / min, and a notable difference in ED was observed, but MMAD was not significant (p>0.05). The high resistance RS00 device showed inhalation flow rate independence caused by a sufficient turbulent flow regime between 60 and 30 L / min (Dal Negro RW, 2015). Thus, the low resistance RS00 device performs better in terms of aerosol properties, which may vary significantly between individual patients, resulting in efficiency variability. However, for the high resistance RS01 device, no flow rate independence was observed between 60 and 30 L / min, confirming that the smaller holes of the RS00 device contribute to the aerosolization of voriconazole nanoaggregates.

[0133] G. By different dose loading The bulk density of voriconazole nanoaggregates before conditioning is typically around 30 mg / cm, regardless of the solids loading (1–3% w / v) of the solution before freezing using TFF. 3 However, the bulk density can be increased to 100 mg / cm3 by conditioning or by applying external physical shear stress. 3The voriconazole nanoaggregates were prepared in a bulk density of approximately 60 mg / cm 3 The effect of conditioning and powder loading level was evaluated in #3 size HPMC capsules. Since the volume of #3 capsules is 0.3 mL, the maximum amount of voriconazole nanoagglomerates that can be inserted into the capsule is approximately 20 mg after conditioning. Therefore, the aerosol performance of voriconazole nanoagglomerates was evaluated in the dose range of 10-20 mg per capsule. High resistance RS00 and RS01 devices were used with a flow rate of 60 L / min, and Tukey-Kramer HSD tests were performed to compare the results between different powder levels.

[0134] The high resistance RS00 device did not show significant differences (p>0.05) in FPF (% of metered dose), FPF (% of delivered dose), and MMAD, but performance was different between 10 mg and 20 mg when using the high resistance RS01 device (p<0.05). The consistency in aerosol performance using the high resistance RS00 device may be a result of small holes that may help aerosolize particles with a narrow distribution in the case of voriconazole nanoagglomerates.

[0135] 3. Materials and Methods A. Material 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) was used, and Pearlitol® PF, pyrogen-free mannitol, was donated by Roquette America Inc. (Geneva, IL).

[0136] B. Preparation of Powder Formulations 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 give 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. For small scale, a 10 mL syringe with a syringe needle (18 gauge) was used to deliver the solution onto the drum. For large scale processes, a Masterflex® L / S® peristaltic pump (Cole-Parmer, Vernon Hills, IL) with Masterflex® L / S® High-performance Precision Platinum-Cured Silicon pump tubing (size 16, Cole-Parmer, Vernon Hills, IL) was used to pump the solution onto the drum at a flow rate of 25 mL / min. During the freezing process, frozen samples were collected in stainless steel freeze-dryer trays filled with liquid nitrogen and transferred to a −80°C freezer to remove excess liquid nitrogen before transferring the samples to the freeze-dryer. The solvent was sublimated and the samples were dried using a VirTis Advantage 2.0 or VirTis Advantage Pro shelf freeze-dryer (VirTis Company Inc., Gardiner, NY). During the primary drying process, the shelf was maintained at −40°C for 20 h, the shelf temperature was linearly increased to 25°C over 20 h, and then maintained at 25°C for 20 h. Secondary drying was performed at 25°C for 20 h. Pressure was maintained at 100 mTorr during the freeze-drying process.

[0137] CX-ray powder diffraction (XRPD) The crystallinity of the powders was identified using X-ray diffraction (MiniFlex 600, Rigaku Corporation, Tokyo, Japan) measuring from 5 to 40° 2θ (0.02° step, 2° / min, 40 kV, 15 mA).

[0138] D. Scanning Electron Microscopy (SEM) A SEM (Zeiss Supra 40V SEM, Carl Zeiss, Heidenheim an der Brenz, Germany) was used to measure the surface morphology of powder samples and to identify nanoparticles after aerosolization. To identify surface morphology, an aliquot of powder was placed on carbon tape and sputter-coated with a 20 nm thick layer of 60 / 40 Pd / Au before being imaged. To measure nanoparticles, 1–2 mg of powder was placed in a DP4 insufflator (Penn-Century Inc., Wyndmoor, PA) and aerosolized using a 3 mL syringe onto a 380 μm single-side polished P-type silicon wafer, which was sputter-coated with a 5 nm thick layer of 60 / 40 Pd / Au before being imaged.

[0139] E. Atomic Force Microscopy (AFM) To obtain images of nanoaggregates, an Asylum MFP-3D AFM (Oxford Instruments, Oxfordshire, UK) equipped with a gold-coated MikroMasch Hi'Res-C15 / Cr-Au cantilever (nanoWorld AG, Neuchâtel, Switzerland) was used. The cantilever has a resonant frequency of 325 kHz, a force constant of 40 N / m, and a typical tip radius of 1 nm. A DP4 insufflator (Penn-Century Inc., Wyndmoor, PA) was utilized to attach the powder to a silicon wafer. 1–2 mg of powder was placed in the insufflator, and a 3 mL syringe was used to aerosolize the powder onto a 380 μm single-sided polished P-type silicon wafer. After loading the powder, excess powder that did not strongly adhere to the wafer was blown off with compressed nitrogen gas. Topography was performed in tapping mode with a scan rate of 1.00 Hz. Other values ​​of the AFM were optimized for each sample. Images were collected using a resolution of 512 × 512 and processed with Gwyddion software (64-bit Windows version 2.50) ( Necas and Klapetek, 2012 ).

[0140] F. Powder Conditioning 1.3 g of powder was added to a 60 mL Pyrex bottle. The bottle was rotated at 60 rpm for 30 minutes to shear the powder and stored in a desiccant at room temperature.

[0141] G. Aerodynamic particle size distribution analysis The aerodynamic properties of the powders were measured by a Next Generation Pharmaceutical Impactor (NGI) (MSP Corporation, Shoreview, MN) equipped with a Critical Flow Controller (Model TPK, MSP Corporation, Shoreview, MN) and a High Capacity Pump (Model HCP5, MSP Corporation, Shoreview, MN). Approximately 5-20 mg of powder formulation was inserted into a #3 HPMC capsule (Vcaps®plus, Capsugel®, Morristown, NJ) and dispersed into the NGI through the USP induction port by either a Plastiape RS01 or RS00 DPI at a total airflow of 4 L. No pre-separator was used. A 1.5% (w / v) solution of polysorbate 20 in methanol was applied to coat the collection plate of the NGI and allowed to dry for 20 min before use. After dispersion, the powder was extracted using a mixture of water and acetonitrile (50:50 v / v) containing 0.1% (v / v) TFA and analyzed for voriconazole content using HPLC. MMAD, geometric standard deviation (GSD), and fine particle fraction (FPF) were calculated using Copley Inhaler Testing Data Analysis Software (CITDAS) version 3.10 (Copley Scientific, Nottingham, UK).

[0142] H. High Performance Liquid Chromatography (HPLC) 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 using 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 confirmed using a wavelength of 254 nm. A linearity study of the standard curve between voriconazole concentrations of 62.5 ng / mL and 500 μg / mL was performed with an injection volume of 7 μL.

[0143] I. Brunauer-Emmett-Teller (BET) specific surface area (SSA) analysis A Monosorb™ Rapid Surface Area Analyzer Model MS-21 (Quantachrome Instruments, Boynton Beach, FL) was used to measure SSA. Powder formulation samples were degassed using 20 psi nitrogen gas for 24 hours at ambient temperature. A mixture of nitrogen and helium (30:70 v / v) was used as the adsorbent gas.

[0144] J. Thermogravimetric analysis (TGA) TGA was performed to measure the moisture content of the powder formulations. A Mettler Thermogravimetric Analyzer, TGA / DSC Model 1 (Columbus, OH) was used. Approximately 2–5 mg of sample was placed in an alumina crucible (Mettler-Toledo, Columbus, OH) and covered with a crucible lid. The crucible was heated from 25 °C to 150 °C at a ramp rate of 5 °C / min. The moisture content of the sample was calculated by comparing the loss of sample weight between 25 °C and 125 °C.

[0145] K. Photographs of freezing at various temperatures To observe the difference in freezing rate among various treatment temperatures by TFF, the freezing process was photographed by a Canon DSLR camera, EOS Rebel SL1 (Canon USA, Melville, NY) equipped with an 18-55 mm IS STM lens (Canon USA, Melville, NY) at a frame rate of 60 frames / s and a resolution of 1280 × 720. The photographed images were cropped to approximately 200 × 200 to show only the sample.

[0146] L. Statistical analysis A Student's t-test was applied to determine whether the aerodynamic properties were statistically different. A p-value less than 0.05 was considered as significant. JMP® 10.0.0 was applied to calculate the p-values ​​of the data.

[0147] All compositions and methods disclosed and claimed herein can be manufactured and carried out without undue experimentation in light of this disclosure.Although the compositions and methods of this disclosure have been described in terms of preferred embodiments, it will be apparent to those skilled in the art that modifications may be made to the methods described herein, and to the steps or sequence of steps of the methods, without departing from the concept, spirit and scope of this disclosure.More specifically, it will be apparent that certain agents that are both chemically and physiologically related may be substituted for the agents described herein to achieve the same or similar results.All such similar substitutes and modifications that are apparent to those skilled in the art are deemed to be within the spirit, scope and concept of this disclosure, as defined by the appended claims.

[0148] References The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference. TIFF2024170583000010.tif197150TIFF2024170583000011.tif231151TIFF2024170583000012.tif231151

Claims

1. A pharmaceutical composition for treating or preventing an infectious disease in a patient, the pharmaceutical composition comprising: (A) a therapeutic agent that is an antifungal agent, and (B) an excipient that is an amino acid or a sugar alcohol and that constitutes less than about 10% by weight of the pharmaceutical composition; Including, the pharmaceutical composition is formulated as a nanoaggregate comprising nanoparticles of the therapeutic agent, the surface of the nanoparticles of the therapeutic agent comprising distinct domains of the excipient, the distinct domains of the excipient reducing the contact area between the nanoparticles of the therapeutic agent; The pharmaceutical composition.

2. The pharmaceutical composition of claim 1, wherein the infection is in the lungs.

3. A pharmaceutical composition described in either claim 1 or 2, wherein the infection is caused by a fungus.

4. The pharmaceutical composition described in any one of claims 1 to 3, wherein the therapeutic agent is an azole antifungal agent.

5. A pharmaceutical composition described in any one of claims 1 to 4, wherein the therapeutic agent is voriconazole.

6. A pharmaceutical composition according to any one of claims 1 to 5, administered to the patient by inhalation.

7. A pharmaceutical composition according to any one of claims 1 to 6, administered to the patient via an inhaler.

8. 8. The pharmaceutical composition of claim 1, wherein the excipient comprises from about 6% w / w to about 2% w / w of the pharmaceutical composition.

9. The pharmaceutical composition of claim 1, wherein the excipient is a sugar alcohol.

10. The pharmaceutical composition of claim 1, wherein the sugar alcohol is mannitol.

11. The pharmaceutical composition of claim 1, wherein the excipient is an amino acid.

12. The pharmaceutical composition of any one of claims 1 to 8 and 11, wherein the amino acid is leucine.

13. A pharmaceutical composition comprising: (A) voriconazole, and (B) an excipient that is an amino acid or a sugar alcohol and that constitutes less than about 10% by weight of the pharmaceutical composition; Including, The pharmaceutical composition is formulated as a nanoaggregate comprising nanoparticles of voriconazole, the surface of the nanoparticles of voriconazole comprising distinct domains of the excipient, the distinct domains of the excipient reducing the contact area between the nanoparticles of voriconazole. The pharmaceutical composition.

14. The pharmaceutical composition of claim 13, wherein the amino acid is leucine.

15. The pharmaceutical composition of claim 13, wherein the sugar alcohol is mannitol.

Images