Dry powder inhalation preparations and their use for therapeutic treatment of the lungs

Abstract

To provide a dry powder inhalation formulation and its use for therapeutic treatment of lungs.SOLUTION: A dry powder inhalation formulation comprises at least one active pharmaceutical ingredient (API) and a lipid matrix comprising at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin, and their mixture. The lipid matrix is hydrogenated castor oil. The ratio between the at least one API and the hydrogenated castor oil is from 10 / 90 to 88 / 12.SELECTED DRAWING: Figure 2

Description

Detailed description of the invention

[0001] The present invention relates to a dry powder inhalation formulation and its use for therapeutic treatment of the lungs.

[0002] Dry powder inhalation (DPI) therapy is currently used primarily to deliver many treatments to the lungs, such as for asthma, and much effort has been made to optimize dry powder inhalation formulations to prolong the retention of active pharmaceutical ingredients (APIs) in the lungs.

[0003] Inhalation allows for the direct administration of high doses of drugs to the lungs without prior distribution within the body. This not only allows for the use of smaller amounts of active pharmaceutical ingredients (APIs) that are likely to be consumed within the body, but also allows for reduced toxicity through direct in situ administration when API toxicity is a concern.

[0004] Furthermore, because the lungs feature a vast absorption surface, inhalation is a promising route of administration for delivering APIs into the systemic circulation.

[0005] However, conventional immediate-release dry powder inhalation (DPI) formulations can nevertheless lead to too short a retention period in the lungs. This is due to multiple mechanisms of clearance for exogenous inhaled particles (i.e., clearance by mucocilia and macrophages) and the rapid dissolution of drug particles, which leads to rapid absorption into the systemic circulation. Therefore, this short retention period necessitates multiple doses, leading to poor patient adherence. Immediate API release can also be a cause of poor local tolerance due to the rapid dissolution of drug particles and the high peak concentration of API in the pulmonary fluid after lung deposition.

[0006] For a sustainable retention profile of the API in the lungs, specific excipients that prolong lung retention have been identified.

[0007] Therefore, further development of dry powder inhalation formulations is necessary to increase the effectiveness of lung treatment.

[0008] In that context, for example, a controlled-release cisplatin-based DPI formulation with high drug load and high particulate fraction was developed (Levet et al, Int J Pharm 2016). This formulation exhibited controlled release and long-term lung retention capacity, leading to low systemic distribution (Levet et al, Int J Pharm 2017).

[0009] To this end, Levet et al. are working on a specific formulation for a single antitumor agent (i.e., cisplatin) that includes a PEGylated excipient to avoid premature elimination by the lung epithelial defense mechanism. Levet et al. are also comparing different carriers, such as polymers and lipid matrices, to allow for longer residence times in the lungs, thereby increasing the efficiency of inhalation.

[0010] In particular, the lipid matrix described in the study by Levet et al. contains tristearin, which was identified as a promising candidate for providing the lipid matrix.

[0011] Furthermore, even lipid-derived excipients for forming the matrix are preferred over polymers, but due to toxicity reasons, only a few substances capable of forming a lipid matrix are acceptable and approved for pharmaceutical compositions for inhalation compared to other routes of therapeutic treatment (Pilcer et Amighi, Int J Pharm 2010).

[0012] Unfortunately, although tristearin has been identified as a promising candidate, its market availability as a pharmaceutical compound is quite limited.

[0013] The present invention aims to address at least some of these obstacles by providing a dry powder inhalation formulation comprising, for example, monotherapy or polytherapy, at least one active pharmaceutical ingredient (API) and a lipid matrix comprising at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof.

[0014] Surprisingly, at least one triglyceride selected from the group including monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof, which are available in pharmaceutical grade, has been identified as being able to form a lipid matrix with an API (active pharmaceutical ingredient) and exhibiting an increased pulmonary deposition rate in the lungs. The latter can be used for pulmonary inhalation.

[0015] In fact, monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof are compounds that have the following physicochemical properties: when measured using the shaking flask method, they have a Log P value greater than 5, preferably 14-25, more particularly 18-25, preferably about 20; and / or have a melting point temperature of 40°C or higher, preferably 60°C or higher, more preferably 75°C or higher.

[0016] This makes it possible to manufacture inhalation formulations in the form of a dry powder, rather than an oil phase or a sticky paste, for proper use in dry powder inhalers.

[0017] Furthermore, according to the present invention, it has been identified, against all expectations, that by changing the ratio between the amount of API and the amount of at least one triglyceride selected from the group including monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof, it is possible to increase the pulmonary deposition rate of API and modulate the solubility profile of API compared to other conventional triglycerides such as tristearin. This was not the case with conventional triglycerides.

[0018] Preferably, the dry powder inhalation formulation according to the present invention has a weight ratio of API / triglyceride between the at least one API and the at least one triglyceride of 0.1 / 99.9 to 99.9 / 0.1, preferably 10 / 90 to 88 / 12, preferably 15 / 85 to 85 / 15, preferably 25 / 75 to 75 / 25, more preferably 30 / 70 to 70 / 30, particularly 40 / 60 to 60 / 40, and for example, about 50 / 50.

[0019] Advantageously, in the formulation according to the present invention, the at least one triglyceride is hydrogenated castor oil.

[0020] Castor oil is listed in the literature as a common excipient (see U.S. Patent Application Publication 2005 / 0042178) or a grinding aid (International Publication 2013 / 128283).

[0021] In a preferred embodiment according to the present invention, the dry powder inhalation formulation contains the at least one API in an amount of 0.1 wt% to 98 wt% based on the total weight of the dry powder inhalation formulation. In a first specific embodiment according to the present invention, the active agent is a small chemical molecule having a solubility in alcohol of at least 0.1 w% / v, or at least 0.5 w% / v, more preferably at least 1 w% / v, and particularly at least 5 w% / v. According to this first specific embodiment, the small chemical molecule has a solubility in alcohol of 0.1% w / v or more and is any active pharmaceutical ingredient (API) defined as an alcohol-soluble API in the present invention. Such any active pharmaceutical ingredient (API) is, for example, budesonide, paclitaxel, pemetrexed, itraconazole, voriconazole, clarithromycin, salbutamol, salbutamol sulfate, fluticasone, beclomethasone, mometasone, mometasone furoate, ciclesonide, formoterol, arformoterol, indacaterol, indacaterol maleate, olodaterol, olodaterol hydrochloride, salmeterol, ipratropium bromide, glycopyrronium bromide, tiotropium bromide, umeclidinium bromide, ibuprofen, vancomycin, vancomycin hydrochloride, tetrahydrolipstatin, isoniazid, rifampicin, pyrazinamide, docetaxel, vincristine, vincristine sulfate, etoposide, vinorelbine, gemcitabine, etc.

[0022] More specifically, the dry powder inhalation formulation according to the first specific embodiment contains the at least one API in an amount of 0.1 wt% to 95 wt%, preferably 1 wt% to 90 wt%, more preferably 1 wt% to 85 wt% based on the weight of the dry powder inhalation formulation.

[0023] In a preferred embodiment according to the present invention, the dry powder inhalation formulation is present in lipid matrix particles in which the alcohol-soluble API is dissolved or finely dispersed.

[0024] In a second specific embodiment according to the present invention, the active agent is a small chemical molecule having a solubility in alcohol of less than 0.5 w% / v. In this second specific embodiment, the small chemical molecule has a solubility in alcohol of less than 0.5% w / v, particularly less than 0.1% w / v, more particularly less than 0.01% w / v, and is any API defined as an alcohol-insoluble API. Such any API is, for example, cisplatin, carboplatin, oxaliplatin, pemetrexed disodium, azacitidine, beclomethasone dipropionate, tobramycin, acridinium bromide, and the like.

[0025] More specifically, the dry powder inhalation formulation according to the second specific embodiment contains the at least one API in an amount of 0.1 wt% to 95 wt%, preferably 1 wt% to 90 wt%, more preferably 1 wt% to 85 wt% based on the weight of the dry powder inhalation formulation.

[0026] Therefore, the dry powder inhalation formulation contains the alcohol-insoluble API in an amount of 0.1 wt% to 95 wt%, preferably 1 wt% to 90 wt%, more preferably 1 wt% to 85 wt%, for example 1 wt% to 60 wt%, or 1 to 50 wt% based on the weight of the dry powder inhalation formulation.

[0027] In a preferred embodiment according to the present invention, the dry powder inhalation formulation is present in lipid matrix particles in which the alcohol-insoluble API is dispersed.

[0028] In a third specific embodiment according to the present invention, the active agent is a macromolecule. In this third specific embodiment according to the present invention, the dry powder inhalation formulation contains the at least one API in an amount of 0.1 wt% to 95 wt%, preferably 1 wt% to 90 wt%, more preferably 1 wt% to 85 wt% based on the weight of the dry powder inhalation formulation.

[0029] Therefore, the dry powder inhalation formulation contains the macromolecule in an amount of 0.1 wt% to 95 wt%, preferably 1 wt% to 90 wt%, more preferably 1 wt% to 85 wt%, for example, 1 wt% to 60 wt%, or 1 to 50 wt%, relative to the total weight of the dry powder inhalation formulation.

[0030] In yet another preferred embodiment, the formulation according to the present invention further comprises a long-lasting pulmonary retention excipient such as a PEGylated excipient, or a polysaccharide such as chitosan or dextran.

[0031] Preferably, according to the present invention, the long-term lung retention excipient is a PEGylated excipient, and is present in an amount of 0.1 wt% to 20 wt%, preferably 0.2 to 10 wt%, and more preferably 0.5 to 5 wt%, relative to the total weight of the dry powder inhalation preparation.

[0032] More specifically, according to the present invention, the PEGylated excipient is derived from vitamin E, or from phospholipids such as tocopheryl polyethylene glycol succinate (TPGS) or distearoyl phosphoethanolamine polyethylene glycol 2000 (DSPE-mPEG-2000).

[0033] In yet another preferred embodiment, the formulation according to the present invention further comprises one or more excipients, such as excipients that improve the physicochemical and / or aerodynamic properties of a dry powder for inhalation.

[0034] The term "one or more excipients" refers to compounds selected from the following: sugar alcohols; polyols (such as sorbitol, mannitol, and xylitol); crystalline sugars including monosaccharides (such as glucose and arabinose) and disaccharides (such as lactose, maltose, sucrose, dextrose, trehalose, and maltitol); inorganic salts (such as sodium chloride and calcium carbonate); organic salts (such as sodium lactate, potassium phosphate or sodium phosphate, sodium citrate, and urea); polysaccharides (such as dextran, chitosan, starch, cellulose, hyaluronic acid, and their derivatives); oligosaccharides (such as cyclodextrin and dextrin); titanium dioxide; silicone dioxide; magnesium stearate; lecithin; and amino acids. (Leucine, isoleucine, histidine, threonine, lysine, valine, methionine, phenylalanine, etc.); amino acid derivatives (acesulfame K, aspartame, etc.); lauric acid or derivatives (esters and salts, etc.); palmitic acid or derivatives (esters and salts, etc.); stearic acid or derivatives (esters and salts, etc.); erucic acid or derivatives (esters and salts, etc.); behenic acid or derivatives (esters and salts, etc.); sodium stearyl fumarate; sodium stearyl lactate; phosphatidylcholine; phosphatidylglycerol; natural and synthetic pulmonary surfactants; lauric acid and its salts (sodium lauryl sulfate, magnesium lauryl sulfate, etc.); triglycerides; sugar esters; phospholipids; cholesterol; talc.

[0035] In certain preferred embodiments, the excipient is mannitol, dextran, or lactose.

[0036] In certain preferred embodiments, the excipient is a phospholipid or cholesterol.

[0037] In certain preferred embodiments, the excipient is mannitol, dextran, hyaluronic acid, lactose, phospholipid, or cholesterol.

[0038] In certain preferred embodiments, the excipient is mannitol, dextran, phospholipid, or cholesterol.

[0039] In certain embodiments, the at least one excipient is a carrier.

[0040] In a preferred embodiment of the formulation according to the present invention, the formulation has a geometric particle size distribution (PSD) d of 30 μm or less, preferably 15 μm or less, preferably 10 μm or less, and preferably 5 μm or less. 50 It is a microparticle with a specific shape.

[0041] In a more preferred embodiment of the formulation according to the present invention, the formulation has a geometric particle size distribution (PSD) d of 60 μm or less, preferably 30 μm or less, more preferably 15 μm or less, preferably 10 μm or less, and more preferably 7 μm or less. 90 It is a microparticle with a specific shape.

[0042] In a further preferred embodiment of the formulation according to the present invention, the formulation is in the form of particles having a volume average diameter D[4,3] of 40 μm or less, preferably 20 μm or less, more preferably 15 μm or less, preferably 10 μm or less, and preferably 6 μm or less.

[0043] Preferably, according to the present invention, the improved excipient and / or the excipient is present in an amount of 0.1% w / w to 80 wt%, preferably less than 70 wt%, more preferably less than 60 wt%, and particularly less than 50 wt%, for example less than 50 wt%, based on the total weight of the dry powder inhalation formulation.

[0044] The term “particulate dose” or “FPD” generally refers to the mass of particles with an aerodynamic diameter of less than 5 μm relative to the mass of the nominal dose (i.e., the mass of the dose loaded into the inhalation device).

[0045] Particulate doses or particulate fractions represent fractions of a pharmaceutical formulation that can be deeply inhaled and are theoretically available for pharmacological activity (Dunbar et al, Kona 16: 7-45, 1998).

[0046] In an advantageous embodiment of the present invention, the formulation is in the form of fine particles having an aerodynamic median mass diameter (MMAD) of 6 μm or less, preferably 5 μm or less, and preferably 4 μm or less.

[0047] MMAD refers to the diameter of particles deposited in an impactor such that 50% (w / w) of the particles have a smaller diameter and 50% (w / w) have a larger diameter.

[0048] The term "aerodynamic diameter" or "DAE" of a particle can be defined as the diameter of a sphere of unit density (i.e., density 1) having the same settling velocity in still air as the particle under consideration. "DAE" provides a useful measurement of inhalable particles, taking into account factors that influence their aerodynamic properties. "DAE" can be used to compare particles of different physical sizes, taking into account their density and shape, as well as their geometric size.

[0049] Methods for measuring "dae" are those described in the European or American Pharmacopoeia, using an impactor or impaction device such as a glass impinger, multi-stage liquid impinger (MsLI), Andersen cascade impactor, or next-generation impactor (NGI). These allow for the measurement of the aerodynamic properties of DPI formulations (including MMAD, geometric standard deviation, lung deposition pattern, particulate dose, and particulate fraction) under simulated respiratory conditions.

[0050] The total dose of particles with an aerodynamic diameter of less than 5 μm can be calculated by interpolation from the collection efficiency curve and can be considered as the particulate dose (FPD) or particulate fraction (FPF), expressed as a percentage of the nominal API dose (i.e., the dose contained in the DPI device).

[0051] Preferably, the dry powder inhalation formulation according to the present invention is packaged, for example, in a blister pack or capsule, for use in a dry powder inhaler, or a sealed dry powder inhaler and / or a disposable dry powder inhaler.

[0052] In certain embodiments, the activator is a small chemical molecule having bronchodilating activity, glucocorticoid activity, anti-inflammatory activity, and anti-infective activity (e.g., an antibiotic, an anti-tuberculosis drug, an antifungal drug, or an antiviral drug).

[0053] In another specific embodiment, the small chemical molecules are, for example, budesonide, salbutamol, fluticasone, beclomethasone, mometasone, ciclesonide, formoterol, salbutamol, alformoterol, indacaterol, orodaterol, salmeterol, ipratropium, acridinium, glycopyrronium, tiotropium, unmeclidinium, mometasone, ciclesonide, formoterol, alformoterol, ibuprofen, tobramycin, vancomycin, tetrahydrolipstatin, clarithromycin, isoniazid, rifampin, pyrazinamide, itraconazole, voriconazole, aztreonam, ethambutol, streptomycin, kanamycin, amikacin, colistin, colistinmethanes Any active pharmaceutical ingredient absorbed into the lungs for systemic or topical treatment, such as sodium rufonate, capreomycin, ciprofloxacin, rifapentin, doxycycline, cycloserine E, ethionamide, gatifloxacin, levofloxacin, moxifloxacin, ofloxacin, fosfomycin, p-aminosalicylic acid, tetrasodium denufosol, lancobutide, ribavirin, zanamivir, laminavir, rupintrivir, pentamidine, amphotericin B, posaconazole, isabconazole, capsufungin, micafungin, anidurafungin, iloprost, levothyroxine, their salts, solvates, hydrates, polymorphs, as well as their esters, combinations thereof, analogs, and derivatives.

[0054] In further embodiments, the activator is a macromolecule such as a peptide, protein, antibody, antibody fragment, nanobody, or nucleic acid.

[0055] Preferably, according to the present invention described above, the macromolecule is insulin, proinsulin, synthetic insulin, semi-synthetic insulin, bevacizumab, pembrolizumab, atezolizumab, nivolumab, ipilimumab, Toll-like receptor agonist, ghrelin, IgG monoclonal antibody, small interfering ribonucleic acid (siRNA), dorunase alpha, cyclosporine A, alpha-1 antitrypsin, interleukin antagonist, interferon-α, interferon-β, interferon-γ, interferon-ω, interleukin-2, anti-IgE mAb, catalase, calcitonin, paratyloid hormone, human growth hormone, insulin-like growth factor-I, heparin, rhG-CSF, GM-CSF, Epo-Fc, FSH-Fc, sFc-γ, RIIb, or mRNA.

[0056] In a more preferred embodiment, the activator is an antitumor agent, for example, provided for lung cancer and lung tumors.

[0057] Lung cancer is the cancer with the highest prevalence and mortality rate in the world. In most cases, lung cancer is diagnosed at an advanced stage. Therefore, patients often already have metastases to the lungs or other organs, i.e., extrapulmonary metastases. Treatment usually involves a combination of therapies, including surgery, radiation therapy, chemotherapy, targeted therapy, and immunotherapy.

[0058] Chemotherapy is used in up to 60% of lung cancer patients, primarily in advanced stages of the disease. Currently, chemotherapy is administered intravenously, via infusion, or orally—that is, through systemic routes of administration; i.e., systemic chemotherapy. Chemotherapy causes severe systemic toxicity due to (i) the widespread distribution of chemotherapy drugs within the body and (ii) a lack of selectivity for cancer cells. Consequently, oncologists have a great need for new, more efficient, and tolerable treatment approaches.

[0059] In reality, chemotherapy merely involves finding the optimal dose of antitumor agent to administer by minimizing the irreversible toxic and unpleasant side effects caused by the chemotherapy to the greatest extent possible.

[0060] Therefore, highly complex treatment schemes are expected to very frequently combine different therapeutic procedures and activities (e.g., surgery) with a complex cycle of a first cycle of radiotherapy followed by injections or intravenous infusions of antitumor agents. The injections or intravenous infusions of antitumor agents may be the same or different for each cycle and may be reduced or adapted depending on the side effects identified in the patient's body.

[0061] Each antitumor agent or molecule possesses dose-controlled toxicities (DLTs), such as nephrotoxicity and neurotoxicity, which necessitate the inclusion of a rest period in the treatment scheme. If non-irreversible, the rest period allows the patient's body to recover from the harmful side effects.

[0062] Furthermore, antitumor agents often have a half-life, also known as their half-life. Half-life is the time required for the concentration or amount of a drug in the body to decrease by half. The half-life of antitumor agents is typically between 12 and 36 hours. This half-life can be shortened to represent the time the human body is exposed to the beneficial effects of the antitumor agent.

[0063] However, due to dose-dependent toxicity (DLT) of antitumor agents, the amount of dose administered during certain treatment periods remains limited, and as mentioned above, these doses should be separated from each other by rest or exclusion periods.

[0064] The combination of half-life and dose-dependent toxicity, along with the fact that antitumor agents are administered orally, by injection, or intravenously, results in low concentrations of the antitumor agent efficiently reaching the solid tumor site, limited effectiveness against the tumor itself, and high systemic toxicity to the patient's body.

[0065] Another limitation of antitumor drugs is cumulative dose. Cumulative dose is the total dose resulting from repeated exposure to the same part of the body or the entire body to the antitumor drug.

[0066] Cisplatin is administered intravenously, with a cumulative dose of 300 mg / m² within 4-6 cycles. 2 That is the case.

[0067] Based on these considerations, the researchers of this invention were able to develop more targeted therapies and on-site injections / infusions.

[0068] Inhalation therapy is a different type of therapy and can be distinguished from nebulizer-type inhalers as a different mode of delivery of APIs to the airways. Inhalers can also be of different types, one of which is the dry powder inhaler (DPI).

[0069] Compared to nebulizers, dry powder inhalers (DPIs) are well-suited for chemotherapy. They allow for the administration of high doses of APIs as well as poorly water-soluble compounds (i.e., most chemotherapy agents in cancer). Furthermore, DPIs limit aerosol contamination of the environment due to (i) their activation and driving solely by the patient's inspiratory flow, and (ii) negligible exhaled drug doses. Finally, DPIs can be designed as single-use, disposable devices.

[0070] Therefore, in order to deliver effective anticancer therapy to tumor-affected lungs, it is necessary to develop pulmonary pathways for antitumor drugs and to adequately adapt current inhalation devices and formulations used in clinical trials.

[0071] Extending the retention profile of antitumor agents in the lungs and ensuring moderate absorption offers numerous advantages. However, the lungs have a vast absorption surface area, which leads to rapid systemic absorption of the drugs, making achieving a long-release profile a challenging task even today.

[0072] As previously explained, controlled-release cisplatin-based DPI formulations have been developed to have high drug-carrying capacity and high particulate fraction, and possess controlled release and long-term lung retention capabilities, leading to low systemic distribution (Levet et al, Int J Pharm 2017). According to this literature, in a mouse model of lung cancer, this approach yielded tumor responses comparable to intravenous regimens in vivo at half the dose (1.0 mg / kg vs. 0.5 mg / kg, respectively). While intravenous regimens yielded good results for extrapulmonary metastases, cisplatin inhalation appeared to be more effective against lung tumors.

[0073] DPI formulations composed of paclitaxel-based nanocarriers have also been developed to exhibit increased lung residence time and limited systemic distribution, specifically targeting lung cancer cells by targeting the folate receptor (FR) (Rosiere et al, Int J Pharm 2016; Rosiere et al, Mol Pharm, 2018). FR, particularly FR-α, is overexpressed on the surface of cancer cells in many lung tumors (i.e., over 70% of adenocarcinomas) and is a promising membrane receptor to target in lung cancer. In lung cancer mouse models, FR-targeted inhalation treatment combined with intravenous administration of Taxol® (commercially available paclitaxel) resulted in significantly longer survival rates compared to intravenous administration of Taxol® alone.

[0074] Returning to the main point, while the use of DPI chemotherapy has led to promising preclinical results in terms of pharmacokinetic profile and safety, its efficacy has been quite limited.

[0075] While achieving a long-term release profile for antitumor agents can yield numerous advantages, this challenge is further highlighted in the context of inhaled chemotherapy, as the lungs exhibit a significantly larger absorption surface for rapidly absorbing APIs and delivering them to the systemic circulation.

[0076] Furthermore, because the lungs have a powerful defense mechanism, the elimination of APIs in inhalation therapy is very rapid due to the highly efficient clearance mechanism in the lungs against foreign particles.

[0077] Sebti et al. (Eur J Pharm Biopharm 2006a; Sebti et al., Eur J Pharm Biopharm 2006b) developed solid lipid macroparticles comprising budesonide and a lipid matrix composed of cholesterol, but no delay in the budesonide release profile was observed.

[0078] For the same purpose, Depreter and Amighi (Depreter and Amighi, Eur J Pharm Biopharm (2010) developed insulin microparticles coated with a lipid matrix composed of cholesterol, but only a slight delay in insulin release was observed.

[0079] For the same purpose, as mentioned above, Levet et al. worked to enable longer residence times in the lungs and thereby increase the efficiency of inhaled chemotherapy, and identified a lipid matrix consisting of tristearin and TPGS as a promising candidate.

[0080] Furthermore, even if lipid-derived excipients for matrix formation are preferable to polymers, only a small number of substances capable of forming a lipid matrix are permissible and approved for inhalation pharmaceutical compositions compared to other routes of therapeutic treatment, due to toxicity reasons (Pilcer and Amighi, Int J Pharm 2010).

[0081] Unfortunately, while tristearin-based matrices have been identified as promising, their market availability as pharmaceutical compounds is quite limited. On the other hand, the availability of at least one triglyceride selected from the group including monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof is generally broader.

[0082] Furthermore, although the tristearin-based matrix was not promising for controlling the release profile of voriconazole, the preparation method (i.e., spray drying) and excipient composition were similar to those described by Levet et al. for cisplatin.

[0083] Furthermore, according to the present invention, it has been identified that at least one triglyceride selected from the group limited by the present invention enables a sustained-release profile and long-term retention of chemotherapy in the lungs, as well as a significant increase in lung deposition rate compared to the closest triglyceride, tristearin.

[0084] Preferably, the antitumor agent is cisplatin, carboplatin, oxaliplatin, docetaxel, paclitaxel, pemetrexed, etoposide, or vinorelbine.

[0085] Other embodiments of the dry powder inhalation formulation according to the present invention are described in the appended claims.

[0086] This invention relates to a method for producing a dry powder inhalation preparation according to the present invention. The method includes the following steps: a) A step of mixing one or more APIs in a predetermined amount of at least one triglyceride selected from the group including monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof, with or without a solvent; b) A bottom-up or top-down method, such as spray drying of a suspension or solution, spray coagulation of a solution of the active drug or API in the at least one triglyceride, or extrusion of a physical mixture of the API and the at least one triglyceride, followed by a jet mill, to produce aspirable particles from the mixture.

[0087] For example, the present invention relates to a method for producing a dry powder inhalation formulation according to the present invention. The method comprises the following steps: suspending or solubilizing one or more API powders in a predetermined amount of at least one triglyceride selected from the group including monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof to form a suspension or solution of particles of one or more APIs (e.g., by melting and / or extrusion); and then reducing the particle size of the solution or suspension of one or more APIs obtained after cooling (e.g., by spray solidification) or extrusion at high speed and / or high pressure to obtain the dry powder inhalation formulation.

[0088] In another example, the manufacturing method includes the following steps: a) A step of homogeneously mixing at least one API with at least one triglyceride to form a homogeneous mixture; b) A step of extruding a homogeneous mixture using a (twin-screw) extruder at an appropriate temperature to obtain a homogeneous lipid matrix containing the API; c) A process of cutting the extruded material to obtain coarse pellets / cylinders; d) Optionally, a step of converting the triglycerides into stable polymorphic phases by storing the coarse pellets / cylinders under appropriate storage conditions; e) A step of grinding the pellets using a suitable mill to obtain inhalable particulate matter (DPI) containing the API together with the at least one triglyceride.

[0089] In another example, the present invention relates to a method for producing a dry powder inhalation preparation according to the present invention. The method includes the following steps: a) A step of suspending or solubilizing one or more API powders in a solvent to form a suspension or solution of particles of one or more APIs; b) A step of arbitrarily reducing the particle size of one or more APIs by high-speed and / or high-pressure homogenization under cooling to form a suspension in which the particle size of one or more APIs has been reduced; c) A step of mixing a predetermined amount of at least one triglyceride, selected from the group comprising monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof, with the microcrystalline suspension or solution in a solvent to obtain a mixture of the at least one API and the at least one triglyceride; d) A step of spray-drying a mixture of the at least one API and the at least one triglyceride to obtain the dried powder inhalation formulation.

[0090] In a preferred embodiment of the method according to the present invention, the high speed applied for size reduction is 10,000 to 30,000 rpm, preferably 15,000 to 26,000 rpm, and the time applied is 8 to 15 minutes, preferably 9 to 12 minutes.

[0091] Preferably, according to the present invention, the high pressure for the homogenization process gradually increases from a first pressure of 2,000 to 10,000 psi, preferably 4,000 to 6,000 psi over a predetermined number of pre-grinding cycles of 8 to 12, preferably 9 to 11, to a second pressure of 8,000 to 12,000 psi, preferably 9,000 to 11,000 psi over a predetermined number of pre-grinding cycles of 8 to 12, preferably 9 to 11, and then gradually increases to a third pressure of 18,000 to 24,000 psi, preferably 19,000 to 24,000 psi over a predetermined number of pre-grinding cycles of 18 to 22, preferably 19 to 21.

[0092] In a particularly preferred embodiment of the present invention, the microcrystals of the microcrystalline suspension have a geometric particle size distribution (PSD) d of 30 μm or less, preferably 15 μm or less, preferably 10 μm or less, and preferably 5 μm or less. 50 It has.

[0093] In a more particularly preferred embodiment of the present invention, the microcrystals of the microcrystalline suspension have a geometric particle size distribution (PSD) d of 60 μm or less, preferably 30 μm or less, more preferably 15 μm or less, preferably 10 μm or less, and more preferably 7 μm or less. 90 It has.

[0094] In another particularly preferred embodiment of the present invention, the microcrystals in the microcrystalline suspension have a volume average diameter D[4,3] of 40 μm or less, preferably 20 μm or less, more preferably 15 μm or less, preferably 10 μm or less, and preferably 6 μm or less.

[0095] In an advantageous and preferred embodiment of the present invention, a PEGylated excipient or other excipient is further added.

[0096] Other embodiments of the method according to the present invention are described in the appended claims.

[0097] The present invention further relates to the use of a dry powder inhalation formulation in lung therapy.

[0098] Preferably, the use of the present invention is anticipated to treat local lung diseases: asthma, COPD, lung infections (e.g., patients with cystic fibrosis, aspergillosis, tuberculosis, etc.), or systemic diseases (e.g., diabetes, pain, etc.).

[0099] In a modified version of the present invention, the dry powder inhalation chemotherapy preparation is used in multi-drug therapy for the treatment of lung cancer, such as any lung tumor, lung metastases (e.g., osteosarcoma metastases), small cell lung cancer, or non-small cell lung cancer.

[0100] Advantageously, the multi-therapy comprises one primary therapy selected from the group consisting of intravenous injection or infusion chemotherapy, immunotherapy, tumor resection surgery, surgical resection to remove part or all of the organ having the tumor, curative surgery, radiotherapy, and combinations thereof, and one or more chemotherapy administered by inhalation as an additional therapy.

[0101] The present invention also relates to corresponding treatment methods.

[0102] [Detailed description of the invention] Other features and advantages of the present invention can be obtained from the following non-limiting description and by reference to the examples and drawings.

[0103] In the drawings, Figure 1 shows the FPF values ​​(%) of the comparative example (composition F5 in Levet et al.) and Examples 1 and 2 (mean ± standard deviation, n=2~3).

[0104] Figure 2 shows the cisplatin release profiles from the breathable fractions of the DPI formulations prepared in Examples 1 and 2, compared to a comparative formulation consisting solely of cisplatin microparticles.

[0105] Figure 3 shows the FPF value of Example 10 compared to a comparative formulation consisting only of pemetrexed microparticles (mean ± standard deviation, n=3 and 1, respectively), demonstrating the superiority of Example 10 over conventional DPI formulations in terms of pulmonary deposition.

[0106] Figure 4 shows the release profile of pemetrexed from the breathable fraction of the DPI formulation prepared in Example 10 compared to a comparative formulation consisting only of pemetrexed microparticles (mean ± standard deviation, n=3).

[0107] Figure 5 shows the FPF values ​​(mean ± standard deviation) for Example 13, demonstrating the high pulmonary deposition rate of the insulin-based DPI composition disclosed in the present invention.

[0108] Figure 6 shows the insulin release profile from the breathable fraction of the DPI preparation prepared in Example 13 (up to 240 minutes, mean ± standard deviation, n=2) (A) compared with the comparative preparation (up to 180 minutes) (B) described by Depreter et al.

[0109] Figure 7 shows the cisplatin release profile from the breathable fraction of the DPI formulation prepared in Example 16 compared to a comparative formulation consisting of cisplatin microparticles (mean ± standard deviation, n=3).

[0110] Figure 8 shows the FPF values ​​of Examples 19 and 20 compared to Comparative Examples BUD-TS4 and BUD-TS5 (mean ± standard deviation, n=3). (***) p<0.001, t-test. The superiority of the budesonide composition disclosed in this invention is demonstrated with respect to lung deposition compared to other triglyceride-based budesonide DPI formulations.

[0111] Figure 9 shows the release profile of budesonide from the breathable fraction of a comparative example of micronized budesonide, as well as the DPI formulations prepared by Examples 19, 20, and 21 (n=1), illustrating the controlled release profile of budesonide from the composition of the present invention and demonstrating the possibility of modifying the release profile by adjusting the drug / lipid ratio.

[0112] [Examples] [Example 1. Preparation of a cisplatin dry powder formulation for inhalation n°1]

[0113] In short, the raw material cisplatin microcrystals derived from the raw powder (Shanghai Jinhe Bio-technology Co., Ltd., Shanghai, PRC) were first suspended in 50 mL of isopropanol to a concentration of 5% w / v. The size was reduced at high speed (24000 rpm for 10 minutes) (X620 motor and T10 dispersion shaft, Ingenieurburo CAT M. Zipperer GmbH, Staufen, Germany), followed by 10 preliminary grinding cycles at 5000 psi, then 10000 psi, followed by 20 grinding cycles at 20000 psi, and then high-pressure homogenization (EmulsiFlex-C5 high-pressure homogenizer, Avestin Inc., Ottawa, Canada). A heat exchanger was connected to the homogenization valve, and an F32-MA cooling circulator (Julabo GmbH, Seelbach, The suspension was maintained at -15°C using a (Germany) laser. At the end of the process, aliquots were removed from the suspension, and the particle size distribution (PSD) of the cisplatin microcrystals was measured by laser diffraction (see below).

[0114] Next, hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA), solubilized in heated isopropanol, were added to a microcrystalline suspension to obtain a mixture of cisplatin at a final concentration of 1.32% w / v and hydrogenated castor oil / TPGS (99:1 w / w) at a final concentration of 0.68% w / v. This mixture was spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use.

[0115] The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0116] The geometric PSDs of bulk cisplatin, cisplatin microparticles from a size reduction process, and cisplatin dry powder formulations were measured as suspended and individualized particles. This was performed using a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.). Aliquot measurements of bulk cisplatin and from the size reduction process were performed in cisplatin-saturated isopropanol. Cisplatin dry powder formulations were pre-dispersed, vortexed, and measured in a 0.9% aqueous solution of Poloxamer 407 (BASF, Ludwigshafen, Germany) saturated with cisplatin at 0.1% w / v. PSD was expressed as the median volume diameter d(0.5) (50% of particles are less than the expressed diameter), the volume mean diameter D[4,3], and the percentage of fine particles (percentage of particles less than 5 μm) determined from the cumulative sieving curve. The PSD results are within the range expected by this invention.

[0117] [Example 2. Preparation of a cisplatin dry powder formulation for inhalation, n°2]

[0118] In short, the raw material cisplatin microcrystals derived from the raw powder (Shanghai Jinhe Bio-technology Co., Ltd., Shanghai, PRC) were first suspended in 50 mL of isopropanol to a concentration of 5% w / v. The size was reduced at high speed (24000 rpm for 10 minutes) (X620 motor and T10 dispersion shaft, Ingenieurburo CAT M. Zipperer GmbH, Staufen, Germany), followed by 10 preliminary grinding cycles at 5000 psi, then 10000 psi, followed by 20 grinding cycles at 20000 psi, and then high-pressure homogenization (EmulsiFlex-C5 high-pressure homogenizer, Avestin Inc., Ottawa, Canada). A heat exchanger was connected to the homogenization valve, and an F32-MA cooling circulator (Julabo GmbH, Seelbach, The suspension was maintained at -15°C using a (Germany) laser. At the end of the process, the alicot was removed from the suspension, and the particle size distribution (PSD) of the cisplatin microcrystals was measured by laser diffraction (see below).

[0119] Next, hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA), solubilized in heated isopropanol, were added to a microcrystalline suspension to obtain a mixture of cisplatin at a final concentration of 1.0% w / v and hydrogenated castor oil / TPGS (99:1 w / w) at a final concentration of 1.0% w / v. This mixture was spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0120] The geometric PSDs of bulk cisplatin, cisplatin microparticles from a size reduction process, and cisplatin dry powder formulations were measured as suspended and individualized particles. This was done using a Mastersizer3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.), as described in Example 1.

[0121] [Example 3. Analysis of deposition rate of cisplatin dry powder formulations prepared by Examples 1 and 2]

[0122] The comparative example (composition F5 in Levet et al.) and the formulations from Examples 1 and 2 were analyzed in terms of their particulate fraction values.

[0123] The particulate fraction (FPF) (the proportion of cisplatin-based particles with an aerodynamic diameter (dae) of less than 5 μm to the recovered dose) and the aerodynamic PSD, characterized by the median mass diameter (MMAD), were determined using MsLI (Copley Scientific, Nottingham, UK) (apparatus C) as described in European Pharmacopoeia 8.0.(2014). 20 mg of each DPI formulation (from Comparative Example and Example 2), pre-sieved through a 355 mm stainless steel mesh, was weighed into a size 3 HPMC capsule (Quali-VI, Qualicaps, Madrid, Spain) and deposited into MsLI using an RS.01 dry powder inhaler (RPC Plastiape, Osnago, Italy) mounted on the inhalation port with an adapter (n=3).

[0124] The deposition flow rate of 100 ± 5 L / min was measured using a DFM3 flow meter (Copley Scientific, Nottingham, UK) and obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series to a TPK critical flow controller (Copley Scientific, Nottingham, UK).

[0125] At these flow rates, the cutoff diameters were 10.0, 5.3, 2.4, 1.3, and 0.4 mm between the stages of MsLI. The microorifice collector (MOC) filter (i.e., stage 5) contained a Fluoropore 9cm PTFE membrane with a pore diameter of 0.45 mm bonded to a high-density polyethylene support (Merck Millipore, Darmstadt, Germany). A critical flow rate controller was used to ensure a deposition time of 2.4 seconds at 100 L / min and a critical flow rate of P3 / P2 ratio < 0.5, as required by the European Pharmacopoeia 8.0.(2014).

[0126] Following impaction, a first rinse using 20 mL of pre-filled 0.5% w / v Poloxamer 407 in ultrapure water / isopropanol (60:40 v / v) as the dilution phase, a second rinse using 25 mL of DMF, and a third rinse using a 100.0 mL dilution phase that had been sonicated for 30 minutes were added to the four upper stages of the MsLI. After solubilization with the 100.0 mL dilution phase and sonication for 30 minutes, drug deposition in the capsule, apparatus, induction port, and MOC filter was measured. The impaction mass at each stage was determined by quantification of cisplatin content by effective electrothermal atomic absorption spectrometry (ETAAS) as described by Levet et al. (Levet, Int J Pharm 2016).

[0127] Next, the results were plotted using Copley Inhaler Testing Data Analysis Software 1 (Copley Scientific, Nottingham, UK) to obtain FPFs of less than 5 μm. This was done by interpolating the recovered mass against the cutoff diameter of the corresponding stage. FPFs were expressed as a percentage of the nominal dose.

[0128] Figure 1 shows the FPF values ​​(%) of the comparative example (composition F5 in Levet et al.) and Examples 1 and 2 (mean ± standard deviation, n=2-3), demonstrating the superiority of the cisplatin composition disclosed in this invention compared to other triglyceride-based cisplatin DPI formulations in terms of lung deposition.

[0129] [Example 4. Analysis of the dissolution rate of cisplatin from the cisplatin dry powder formulations prepared by Examples 1 and 2]

[0130] The solubility properties of the DPI formulation were demonstrated by applying the method described by Levet et al. (Int J Pharm 2016). This method is derived from the paddle-over-disk method from USP39 using a Type V dissolution device modified for transdermal patches. The cisplatin release profile was determined from the total breathable fraction (dae ≤ 5 μm) of the DPI formulation, selected using a Fast Screening Impactor (FSI, Copley Scientific, Nottingham, UK). The appropriate mass of each DPI formulation, corresponding to the deposition amount of 3 mg of cisplatin, was weighed into size 3 HPMC capsules (Quali-VI Qualicaps, Madrid, Spain). Next, this was deposited onto a 0.45 mm pore size Fluoropore® hydrophobic PTFE membrane filter (Merck Millipore, Darmstadt, Germany) with an FSI (2.4 s, 100 L / min) equipped with a corresponding pre-separator insert, using an RS.01 DPI instrument (RPC Plastiape). Then, the Fluoropore filter with the deposited powder facing upwards was covered with an Isopore® 0.4 mm hydrophilic polycarbonate filter (Merck-Milipore, Germany) and fixed onto a watch glass-PTFE disc assembly (Copley, Nottingham, UK) equipped with clips and a PTFE mesh screen. The disc assembly was then immersed in a dissolution vessel of an AT7 dissolution apparatus (Sotax AG, Aesch, Switzerland) containing 400 mL of modified simulated lung fluid (mSLF) (Son and McConville, 2009) (a medium mimicking a lung electrolyte and surfactant composition).

[0131] The dissolution test was performed under sink conditions of 37±0.2℃ and pH 7.35±0.05. The paddle, set to 25±2mm between the center of the blade and the center of the disk assembly, was set to a rotation speed of 50±4rpm. A sample volume of 2.0 mL was passed through a cellulose acetate syringe filter (VWR, Leuven, Belgium) with a pore size of 0.22 mm, filtered for a preset time between 2 minutes and 24 hours, and replaced with 2.0 mL of free preheated mSLF.

[0132] At the end of the dissolution assay, the disk assembly was emptied into a dissolution vessel and sonicated for 30 minutes to establish a 100% cisplatin dissolution rate.

[0133] Figure 2 shows the cisplatin release profiles from the breathable fractions of the DPI formulations prepared in Examples 1 and 2, compared to a comparative formulation consisting solely of cisplatin microparticles.

[0134] [Example 5. Preparation of dried insulin powder formulation n°1]

[0135] First, the insulin was suspended in isopropanol (2% w / v) and sonicated for 10 minutes in a 40 kHz Branson 2510 bath to ensure powder dispersion. The particle size was then reduced using an EmulsiFlex-C5 high-pressure homogenizer (Aves-tin Inc., Ottawa, Canada). A preliminary low-pressure homogenization cycle was then performed on the insulin suspension to further reduce the particle size (10 cycles at 7000 PSI and 10 cycles at 12,000 PSI). Finally, HPH was applied for 30 cycles at 24,000 PSI. These cycles were performed by directly recirculating the treated suspension into a closed-loop sample tank. Since HPH causes a rise in sample temperature (a 30°C rise after 20 cycles at 24,000 PSI), all operations were performed using a heat exchanger placed in front of the homogenization valve to maintain the sample temperature at 5 ± 1°C.

[0136] At the end of the process, aliquots were removed from the suspension, and the particle size distribution (PSD) of insulin microcrystals was measured by laser diffraction (see below).

[0137] Next, hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA), solubilized in heated isopropanol, were added to a microcrystalline suspension to obtain a mixture of insulin at a final concentration of 1.0% w / v and hydrogenated castor oil / TPGS (99:1 w / w) at a final concentration of 1.0% w / v. This mixture was spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0138] The geometric PSDs of bulk insulin, insulin microparticles from a size reduction process, and insulin dry powder formulations, when measured as suspended and individualized particles, were within the scope of the present invention. This was done as described in Example 1, using a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.).

[0139] [Example 6. Preparation of dried insulin powder formulation n°2]

[0140] First, insulin was suspended in isopropanol (2% w / v) and sonicated for 10 minutes in a 40 kHz Branson 2510 bath to ensure dispersion of the powder. Next, an EmulsiFlex-C5 high pressure homogenizer (Aves-tin Inc., Ottawa, Canada) was used to reduce the particle size. First, a pre-crushing low pressure homogenization cycle was performed on the insulin suspension to further reduce the particle size (10 cycles at 7000 PSI and 10 cycles at 12,000 PSI). Then, finally, HPH was applied at 24,000 PSI for 30 cycles. These cycles were performed by directly recirculating the treated suspension into the sample tank (closed loop). Since HPH causes an increase in the sample temperature (it rises by 30 °C after 20 cycles at 24,000 PSI), all operations were performed using a heat exchanger placed before the homogenization valve to maintain the sample temperature at 5 ± 1 °C.

[0141] At the end of the process, an aliquot was removed from the suspension and the particle size distribution (PSD) of the insulin microcrystals was measured by laser diffraction (see below).

[0142] Next, hardened castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA) solubilized in heated isopropanol were added to the microcrystal suspension to obtain a mixture of 1.5% w / v insulin and 0.5% w / v hardened castor oil / TPGS (99:1 w / w), and spray dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for humans. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70 °C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and drying air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and the relative humidity was maintained at 50% HR during spray drying.

[0143] The geometric PSDs of bulk insulin, insulin microparticles from a size reduction process, and insulin dry powder formulations, when measured as suspended and individualized particles, were within the scope of the present invention. This was done as described in Example 1, using a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.).

[0144] [Example 7. Preparation of a budesonide dry powder formulation for inhalation, n°1]

[0145] First, budesonide (1% w / v) was solubilized in isopropanol under magnetic stirring. Next, hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA), solubilized in heated isopropanol, were added to a microcrystalline suspension to obtain a mixture of budesonide at a final concentration of 1.0% w / v and hydrogenated castor oil / TPGS (99:1 w / w) at a final concentration of 1.0% w / v. This mixture was spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0146] The geometric PSD of the bulk budesonide dry powder formulation, measured as suspended and individualized particles, was within the scope of the present invention. This was performed as described in Example 1, using a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.).

[0147] [Example 8. Preparation of a budesonide dry powder formulation for inhalation, n°2]

[0148] First, budesonide (1.5% w / v) was solubilized in isopropanol under magnetic stirring. Next, hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA), solubilized in heated isopropanol, were added to a microcrystalline suspension to obtain a mixture of budesonide at a final concentration of 1.5% w / v and hydrogenated castor oil / TPGS (99:1 w / w) at a final concentration of 0.5% w / v. This mixture was spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0149] The geometric PSD of the bulk budesonide dry powder formulation, measured as suspended and individualized particles, was within the scope of the present invention. This was performed as described in Example 1, using a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.).

[0150] [Example 9. Preparation of budesonide dry powder formulation n°3]

[0151] In a Turbula® mixer (Willy A. Bachofen AG, Muttenz, Switzerland), 10 g of pulverized budesonide, 9.9 g of hydrogenated castor oil, and 0.1 g of TPGS were homogeneously blended. The homogeneous blend was then extruded at an appropriate temperature through a twin-screw extruder (Process-11, Thermo Fischer Scientific, Massachusetts, USA) to obtain a homogeneous lipid matrix. The extruded material was then cut to obtain coarse pellets, which were placed in an incubator under appropriate storage conditions to convert the lipid matrix into a stable polymorphic phase. Finally, the pellets were ground using a jet mill (with appropriate pellet feed rate, injection pressure, and grinding pressure) to obtain fine particles for human inhalation.

[0152] [Example 10. Preparation of pemetrexed dry powder formulation for inhalation n°1]

[0153] In short, microcrystalline pemetrexed disodium (heptahydrate form) derived from the raw material (Carbosynth Limited, Berkshire, United Kingdom) was first suspended in 50 mL of isopropanol to a concentration of 1% w / v in the presence of 0.05% w / v TPGS (Sigma-Aldrich, St-Louis, USA). The size was reduced at high speed (24,000 rpm for 10 minutes) (X620 motor and T10 dispersion shaft, Ingenieurburo CAT M. Zipperer GmbH, Staufen, Germany), followed by 20 grinding cycles at 25,000 psi and high-pressure homogenization (EmulsiFlex-C3 high-pressure homogenizer, Avestin Inc., Ottawa, Canada). A heat exchanger was connected to the homogenization valve, and the temperature was maintained at +5°C using an F32-MA cooling circulator (Julabo GmbH, Seelbach, Germany). At the end of the process, aliquots were removed from the suspension, and the particle size distribution (PSD) of the pemetrexed microcrystals was measured by laser diffraction (see below).

[0154] Next, 1% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany) was solubilized in a heated (50°C) microcrystalline suspension and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and drying air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0155] The geometric PSDs of bulk pemetrexed, pemetrexed microparticles from a size reduction process, and pemetrexed dry powder formulations were measured as suspended and individualized particles and were within the scope of the present invention. This was done as described in Example 1, using a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.).

[0156] [Example 11. Analysis of the deposition rate of the pemetrexed dry powder formulation prepared according to Example 10]

[0157] The formulations prepared in Example 10 were analyzed in terms of their in vitro lung deposition patterns and FPF values.

[0158] FPF (the ratio of pemetrexed-based particles with an aerodynamic diameter (dae) of less than 5 μm to the recovered dose) and aerodynamic PSD, characterized by MMAD, were determined using NGI (Copley Scientific, Nottingham, UK) (apparatus E) as described in European Pharmacopoeia 8.0.(2014). 20 mg of the DPI formulation (according to Example 10, and a comparative formulation consisting only of pemetrexed microparticles), pre-sieved through a 355 mm stainless steel mesh, was weighed in a size 3 HPMC capsule (Quali-VI, Qualicaps, Madrid, Spain) and deposited in NGI using an RS.01 dry powder inhaler (RPC Plastiape, Osnago, Italy) mounted on the inhalation port with an adapter (n=3).

[0159] The deposition flow rate of 100 ± 5 L / min was measured using a DFM3 flow meter (Copley Scientific, Nottingham, UK) and obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series to a TPK critical flow controller (Copley Scientific, Nottingham, UK).

[0160] At these flow rates, the cutoff diameters between the NGI stages were 6.12, 3.42, 2.18, 1.31, 0.72, 0.40, and 0.24 μm. A critical flow controller was used to ensure a deposition time of 2.4 seconds at 100 L / min and a critical flow rate of P3 / P2 ratio < 0.5, as required by the European Pharmacopoeia 8.0 (2014).

[0161] After impaction, the pemetrexed mass deposited on the capsule, apparatus, lead port, pre-separator, 7 stages, and MOC of NGI was collected using ultrapure water / DMF (30:70 v / v) as the dilution phase and sonicated for 30 minutes. The impaction mass at each stage was determined by quantification of the pemetrexed content by effective HPLC. The chromatography system (HP 1200 series, Agilent Technologies, Diegem, Belgium) was equipped with four pumps, an autosampler, and a diode array detector. Separation was performed using a reversed-phase Hypersil Gold C18 column (5 mm, 250 mm × 4.6 mm) (Thermo Fisher Scientific, Waltham, USA). The mobile phase consisted of ultrapure water / acetonitrile (86:14) acidified with 0.4% formic acid and delivered at a flow rate of 1 mL / min. Quantification was performed at 256 nm. The injection volume was 20 μL, the temperature was set to 30°C, and the analysis execution time was 15 minutes.

[0162] Next, the results were plotted using Copley Inhaler Testing Data Analysis Software 1 (Copley Scientific, Nottingham, UK) to obtain FPFs of less than 5 μm. This was done by interpolating the recovered mass against the cutoff diameter of the corresponding stage. FPFs were expressed as a percentage of the nominal dose.

[0163] Figure 3 shows the FPF value of Example 10 compared to a comparative formulation consisting only of pemetrexed microparticles (mean ± standard deviation, n=3 and 1, respectively), demonstrating the superiority of Example 10 over conventional DPI formulations in terms of pulmonary deposition.

[0164] [Example 12. Analysis of the dissolution rate of pemetrexed from the pemetrexed dry powder formulation prepared according to Example 10]

[0165] The solubility properties of the DPI formulation were demonstrated by applying an adapted method described by Pilcer et al. (J Pharm Sci 2013). A solubility system (Copley Scientific, Nottingham, UK) specifically developed for investigating the DPI release profile was used, adapted from the "Paddle over Disc" method (Eur. Ph. 7). To investigate the release profile of particles deposited in the lung, fractionation of the pemetrexed formulation was first performed using NGI. The stage 3 cup was selected to have a removable disc insertion section for particle collection. Particularly interesting, at the selected inhalation rate (100 L / min for 2.4 seconds), the cutoff diameter of stage 3 was in the range of 2.18–3.42 μm, allowing for the selection of particles targeting the lung. Capsules containing an appropriate amount of the formulation according to Example 10 were weighed, and approximately 6 mg of pemetrexed was collected in stage 3. Next, the disc insertion area is covered with a polycarbonate membrane (pore size 0.4 μm) (Merck Millipore), and a paddle dissolution apparatus (Erweka DT6; ERWEKA) is filled with 400 mL of mSLF (Son and McConville, 2009) (a culture medium mimicking a pulmonary electrolyte and surfactant composition). I placed my job at GmbH, Heusenstamm, Hesse, Germany.

[0166] The dissolution test was performed under sink conditions of 37±0.2℃ and pH 7.35±0.05. The paddle, set to 25±2mm between the center of the blade and the center of the disk assembly, was set to a rotation speed of 50±4rpm. A sample volume of 2.0 mL was passed through a cellulose acetate syringe filter (VWR, Leuven, Belgium) with a pore size of 0.22 mm, filtered for a preset time between 2 minutes and 24 hours, and replaced with 2.0 mL of free preheated mSLF.

[0167] At the end of the dissolution assay, the disk assembly was emptied into a dissolution vessel and sonicated for 30 minutes to establish a 100% pemetrexed dissolution value.

[0168] Figure 4 shows the release profile of pemetrexed from the breathable fraction of the DPI formulation prepared in Example 10 compared to a comparative formulation consisting only of pemetrexed microparticles (mean ± standard deviation, n=3).

[0169] The two dissolution profiles were compared using a similarity factor f2 (Shah et al, Pharm Res 1998). The curves were significantly different (f2<50). Furthermore, the cumulative release values ​​at all time points from Example 10 were significantly lower (p<0.05, t-test) compared to those from pemetrexed microcrystals, for example, 53±9% and 97.9±0.9% at 1 hour, respectively (p<0.01), which indicates a controlled release profile of pemetrexed from the composition of the present invention.

[0170] [Example 13. Preparation of dried insulin powder formulation n°3]

[0171] First, insulin (Sigma-Aldrich) was suspended in isopropanol (1% w / v) using a magnetic stirrer, and the powder was sonicated for 10 minutes in a 40 kHz Branson 2510 bath to ensure dispersion. Then, the particle size was reduced for 30 cycles at 22,000 PSI using an EmulsiFlex-C3 high-pressure homogenizer (Avestin Inc., Ottawa, Canada). These cycles were performed by directly recirculating the treated suspension into a sample tank (closed loop). Since high heat per pound (HPH) causes an increase in sample temperature, all operations were performed using a heat exchanger placed in front of the homogenization valve to maintain the sample temperature at 5 ± 1°C.

[0172] At the end of the process, aliquots were removed from the suspension, and the particle size distribution (PSD) of insulin microcrystals was measured by laser diffraction (see below).

[0173] Next, 1% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany) was solubilized in a heated (50°C) microcrystalline suspension and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0174] The geometric PSDs of bulk insulin, insulin microparticles from a size reduction process, and insulin dry powder formulations, when measured as suspended and individualized particles, were within the scope of the present invention. This was done as described in Example 1, using a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.).

[0175] [Example 14. Analysis of deposition rate of the insulin dry powder formulation prepared according to Example 13]

[0176] The formulation prepared according to Example 13 was analyzed in terms of its FPF value.

[0177] The Fast Filtration Process (FPF), which is the proportion of insulin-based particles with a dae size less than 5 μm to the recovered dose, was determined using a Fast Screening Impactor (FSI) (Copley Scientific, Nottingham, UK). The FSI employs a two-step separation process, where a first large non-inhalable bolus is captured in a liquid trap, followed by a micro-cutting impaction step at 5 microns (i.e., corresponding to the FPF). A mass of 10 mg of a DPI formulation (according to Example 13), pre-sieved through a 355 mm stainless steel mesh, was weighed in a size 3 HPMC capsule (Quali-VI, Qualicaps, Madrid, Spain) and deposited into the FSI using an RS.01 dry powder inhaler (RPC Plastiape, Osnago, Italy) mounted on the inhalation port with an adapter (n=3).

[0178] The deposition flow rate of 100 ± 5 L / min was measured using a DFM3 flow meter (Copley Scientific, Nottingham, UK) and obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series to a TPK critical flow controller (Copley Scientific, Nottingham, UK). A deposition time of 2.4 seconds at 100 L / min was ensured using the critical flow controller.

[0179] After impaction, the insulin mass deposited on the capsule, apparatus, induction port, pre-separator, and Fluoropore 9cm PTFE membrane with a pore size of 0.45 mm, bound to a high-density polyethylene support (Merck Millipore, Darmstadt, Germany), was collected using 0.01 M HCl as the dilution phase and sonicated for 30 minutes. The impaction mass at each stage was determined by quantification of insulin content by HPLC as described in European Pharmacopoeia 9.2.(2017). FPF was expressed as a percentage of the nominal dose.

[0180] Figure 5 shows the FPF values ​​(mean ± standard deviation) for Example 13, demonstrating the high pulmonary deposition rate of the insulin-based DPI composition disclosed in the present invention.

[0181] [Example 15. Analysis of the dissolution rate of insulin from the insulin dry powder preparation prepared according to Example 13]

[0182] The solubility characteristics of the DPI formulation according to Example 13 were demonstrated by applying an adapted method described by Depreter et al. (Eur J Pharm Biopharm 2012). A solubility system (Copley Scientific, Nottingham, UK) specifically developed for investigating the DPI release profile was used, adapted from the "Paddle over Disc" method (Eur. Ph. 7). To investigate the release profile of particles deposited in the lung, fractionation of the insulin formulation was first performed using NGI. The Stage 3 cup was selected to have a removable disc insertion section for particle collection. Of particular interest, at the selected inhalation rate (100 L / min for 2.4 seconds), the Stage 3 cutoff diameter was in the range of 2.18–3.42 μm, allowing for the selection of particles targeting the lung. Capsules containing an appropriate amount of the formulation according to Example 13 were weighed, and approximately 3 mg of insulin was collected in Stage 3. Next, the disc insertion area was covered with a polycarbonate membrane (pore size 0.4 μm) (Merck Millipore) and placed in a paddle dissolving apparatus (Erweka DT6; ERWEKA GmbH, Heusenstamm, Hesse, Germany) filled with 400 mL of 0.01 mM phosphate buffered saline (pH 7.4).

[0183] The dissolution test was performed under sink conditions of 37±0.2℃ and pH 7.35±0.05. The paddle, set to 25±2mm between the center of the blade and the center of the disk assembly, was set to a rotation speed of 50±4rpm. A 5.0mL sample was taken at a preset time between 2 minutes and 24 hours and replaced with 5.0mL of free preheated PBS.

[0184] At the end of the dissolution assay, the disc assembly was emptied into a dissolution vessel and sonicated for 30 minutes to establish 100% insulin dissolution.

[0185] The samples were lyophilized in the presence of 3% w / v trehalose (Christ Epsilon 1-6). The lyophilized material was dissolved in 500 μL of 0.02N HCl and injected into an HPLC system using the method described in European Pharmacopoeia 9.2 (2017).

[0186] Figure 6 shows the insulin release profile from the breathable fraction of the DPI preparation prepared in Example 13 (up to 240 minutes, mean ± standard deviation, n=2) (A) compared with the comparative preparation (up to 180 minutes) (B) described by Depreter et al.

[0187] The cumulative release values ​​at various time points from Example 13 were lower than those from the two formulations from Depreter et al., for example, at 1 hour they were approximately 60% and 100%, respectively, demonstrating the superiority of the insulin composition disclosed in this invention over both the insulin microcrystals and lipid-coated insulin microcrystals (F1 and F2 in Figure 6, respectively) described by Depreter et al. with respect to the control of insulin release.

[0188] [Example 16. Preparation of cisplatin dried powder formulation n°3 and comparative example]

[0189] First, cisplatin (Umicore, Hanau-Wolfgang, Germany) was suspended in 50 mL of ethanol to a concentration of 5% w / v, reduced in size at high speed (24,000 rpm for 10 minutes) (X620 motor and T10 dispersion shaft, Ingenieurburo CAT M. Zipperer GmbH, Staufen, Germany), and then homogenized under high pressure using an EmulsiFlex-C3 high-pressure homogenizer (Avestin Inc., Ottawa, Canada) (40 cycles at 20,000 PSI). These cycles were performed by directly recirculating the treated suspension into a sample tank (closed loop). Since high heat per second (HPH) causes an increase in sample temperature, all operations were performed using a heat exchanger placed in front of the homogenization valve to maintain the sample temperature at 15 ± 1 °C.

[0190] At the end of the process, aliquots were removed from the suspension, and the PSD of the cisplatin microcrystals was measured by laser diffraction (see below).

[0191] Next, hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA) (or tristearin for comparative examples), solubilized in heated isopropanol, were added to a microcrystalline suspension to obtain a mixture of cisplatin at a final concentration of 2.0% w / v and triglyceride / TPGS (99:1 w / w) at a final concentration of 2.0% w / v. This mixture was spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0192] The geometric PSDs of bulk cisplatin, cisplatin microparticles from a size reduction process, and cisplatin dry powder formulations, when measured as suspended and individualized particles, were within the scope of the present invention. This was done as described in Example 1, using a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) connected to a Hydro MV dispenser equipped with a 40W ultrasonic probe (Malvern Instruments Ltd.).

[0193] [Example 17. Analysis of deposition rate of cisplatin dry powder formulation prepared according to Example 6 16]

[0194] The formulations from Example 16 were analyzed in terms of their in vitro lung deposition patterns and FPF values.

[0195] FPF (percentage of cisplatin-based particles with dae less than 5 μm relative to the recovered dose) and aerodynamic PSD characterized by MMAD were determined using NGI (Copley Scientific, Nottingham, UK) (apparatus E) as described in European Pharmacopoeia 8.0.(2014). 20 mg of the DPI formulation (according to Example 17), pre-sieved through a 355 mm stainless steel mesh, was weighed in a size 3 HPMC capsule (Quali-VI, Qualicaps, Madrid, Spain) and deposited in NGI using an RS.01 dry powder inhaler (RPC Plastiape, Osnago, Italy) mounted on the inhalation port with an adapter (n=3).

[0196] The deposition flow rate of 100 ± 5 L / min was measured using a DFM3 flow meter (Copley Scientific, Nottingham, UK) and obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series to a TPK critical flow controller (Copley Scientific, Nottingham, UK).

[0197] At these flow rates, the cutoff diameters between the NGI stages were 6.12, 3.42, 2.18, 1.31, 0.72, 0.40, and 0.24 μm. A critical flow controller was used to ensure a deposition time of 2.4 seconds at 100 L / min and a critical flow rate of P3 / P2 ratio < 0.5, as required by the European Pharmacopoeia 8.0.(2014).

[0198] After impaction, the cisplatin mass deposited in the capsule, apparatus, induction port, pre-separator, 7 stages, and MOC of NGI was collected using DMF as the dilution phase and sonicated for 30 minutes. The impaction mass at each stage was determined by quantitative determination of cisplatin content using the effective ETAAS method described by Levet et al. (Int J Pharm 2016).

[0199] Next, the results were plotted using Copley Inhaler Testing Data Analysis Software 1 (Copley Scientific, Nottingham, UK) to obtain FPFs of less than 5 μm. This was done by interpolating the recovered mass against the cutoff diameter of the corresponding stage. FPFs were expressed as a percentage of the nominal dose.

[0200] The results obtained were integrated with those obtained in Example 3, demonstrating the superiority of the cisplatin composition disclosed in this invention compared to other triglyceride-based cisplatin DPI formulations with respect to pulmonary deposition.

[0201] [Example 18. Analysis of the dissolution rate of cisplatin from the cisplatin dry powder formulation prepared according to Example 16]

[0202] The solubility properties of the DPI formulation were demonstrated by applying an adapted method described by Pilcer et al. (J Pharm Sci 2013). A dissolution system (Copley Scientific, Nottingham, UK) specifically developed for investigating the DPI release profile was used, adapted from the "Paddle over Disc" method (Eur. Ph. 7). To investigate the release profile of particles deposited in the lung, fractionation of the cisplatin formulation was first performed using NGI. The Stage 3 cup was selected to have a removable disc insertion portion for particle collection. Particularly interesting, at the selected inhalation rate (100 L / min for 2.4 seconds), the Stage 3 cutoff diameter was in the range of 2.18–3.42 μm, allowing for the selection of lung-targeting particles from a capsule filled with an appropriate amount of the formulation according to Example 16, depositing approximately 2 mg of cisplatin in Stage 3. Next, the disc insertion area is covered with a polycarbonate membrane (pore size 0.4 μm) (Merck Millipore), and a paddle dissolution apparatus (Erweka DT6; ERWEKA GmbH) filled with 400 mL of mSLF (Son and McConville, 2009) (a culture medium mimicking a pulmonary electrolyte and surfactant composition) is placed inside. I put it in Heusenstamm, Hesse, Germany.

[0203] The dissolution test was performed under sink conditions of 37±0.2℃ and pH 7.35±0.05. The paddle, set to 25±2mm between the center of the blade and the center of the disk assembly, was set to a rotation speed of 50±4rpm. A sample volume of 2.0 mL was passed through a cellulose acetate syringe filter (VWR, Leuven, Belgium) with a pore size of 0.22 mm, filtered for a preset time between 2 minutes and 24 hours, and replaced with 2.0 mL of free preheated mSLF.

[0204] At the end of the dissolution assay, the disk assembly was emptied into a dissolution vessel and sonicated for 30 minutes to establish a 100% cisplatin dissolution rate.

[0205] Figure 7 shows the cisplatin release profile from the breathable fraction of the DPI formulation prepared in Example 16 compared to a comparative formulation consisting of cisplatin microparticles (mean ± standard deviation, n=3).

[0206] The two dissolution profiles were compared using a similarity factor f2 (Shah et al, Pharm Res 1998). The curves were significantly different (f2<50). Furthermore, the cumulative release values ​​at all time points from Example 16 were significantly lower (p<0.05, t-test) compared to those from cisplatin microcrystals, for example, 57±4% and 76±5% at 4 hours, respectively (p<0.01), which indicates a controlled release profile of cisplatin from the composition of the present invention.

[0207] [Example 19. Preparation of a budesonide dry powder formulation for inhalation, n°4]

[0208] 0.1500% w / v budesonide, 2.8215% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany) (or tristearin for comparative example BUD-TS4), and 0.0285% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm isopropanol (65°C) under magnetic stirring, and then dried in a Mini-Spray Dryer. The DPI formulation for human use was obtained by spray drying using a B-290 (Buchi Labortechnik AG, Flawil, Switzerland). The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0209] [Example 20. Preparation of a budesonide dry powder formulation for inhalation, n°5]

[0210] 0.600% w / v budesonide, 2.376% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany) (or tristearin for comparative example BUD-TS5), and 0.024% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm isopropanol (65°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0211] [Example 21. Preparation of a budesonide dry powder formulation for inhalation, n°6]

[0212] 1,500% w / v budesonide, 1,485% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany), and 0.015% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm isopropanol (65°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and drying air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0213] [Example 22. Analysis of deposition rate of budesonide dry powder formulations prepared by Examples 19 and 20]

[0214] The formulations from Examples 19 and 20 were analyzed in terms of their in vitro lung deposition patterns and particulate fraction values. Comparative examples of formulations 19 and 20, namely BUD-TS4 and BUD-TS5, were prepared using the corresponding identical protocols, with tristearin (TS) (Tokyo Chemical Company, Tokyo, Japan) instead of hydrogenated castor oil.

[0215] FPF (percentage of budesonide-based particles with dae less than 5 μm relative to the recovered dose) and aerodynamic PSD characterized by MMAD, as described in European Pharmacopoeia 8.0.(2014), NGI (Copley Scientific, Nottingham, The determination was made using (UK) (apparatus E). 10 mg of DPI formulations (from Examples 19 and 20, and comparative powders BUD-TS4 and BUD-TS5), pre-sieved through a 355 mm stainless steel mesh, was weighed in size 3 HPMC capsules (Quali-VI, Qualicaps, Madrid, Spain) and deposited in NGI using an RS.01 dry powder inhaler (RPC Plastiape, Osnago, Italy) mounted on the inhalation port with an adapter (n=3).

[0216] The deposition flow rate of 100 ± 5 L / min was measured using a DFM3 flow meter (Copley Scientific, Nottingham, UK) and obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series to a TPK critical flow controller (Copley Scientific, Nottingham, UK).

[0217] At these flow rates, the cutoff diameters between the NGI stages were 6.12, 3.42, 2.18, 1.31, 0.72, 0.40, and 0.24 μm. A critical flow controller was used to ensure a deposition time of 2.4 seconds at 100 L / min and a critical flow rate of P3 / P2 ratio < 0.5, as required by the European Pharmacopoeia 8.0.(2014).

[0218] After impaction, the budesonide mass deposited on the capsule, apparatus, induction port, pre-separator, 7 stages, and MOC of NGI was collected using 0.5% w / v poloxamer 407 in a 60:40 (v / v) mixture of ultrapure water:isopropanol as the dilution phase, and sonicated at 60°C for 30 minutes. The solution was then filtered through a regenerated cellulose Minisart syringe filter (Sartorius Stedim Biotech GmbH, Germany) with a pore size of 0.45 μm. The impaction mass at each stage was determined by quantification of budesonide content by effective HPLC. The chromatography system (HP 1200 series, Agilent Technologies, Diegem, Belgium) was equipped with four pumps, an autosampler, and a diode array detector. Separation was performed using a reversed-phase Alltima C18 column (5 mm, 150 mm × 4.6 mm) (Hichrom, Theale, UK). The mobile phase consisted of phosphate buffer:acetonitrile (65:35 v / v) at pH 3.20 and was delivered at a flow rate of 1 m.5 L / min. Quantification was performed at 245 nm. The injection volume was 100 μL, the temperature was set to 40°C, and the analysis execution time was 22 minutes.

[0219] Next, the results were plotted using Copley Inhaler Testing Data Analysis Software 1 (Copley Scientific, Nottingham, UK) to obtain FPFs of less than 5 μm. This was done by interpolating the recovered mass against the cutoff diameter of the corresponding stage. FPFs were expressed as a percentage of the nominal dose.

[0220] Figure 8 shows the FPF values ​​of Examples 19 and 20 compared to Comparative Examples BUD-TS4 and BUD-TS5 (mean ± standard deviation, n=3). (***) p<0.001, t-test. The superiority of the budesonide composition disclosed in this invention is demonstrated with respect to lung deposition compared to other triglyceride-based budesonide DPI formulations.

[0221] [Example 23. Analysis of the dissolution rate of budesonide from budesonide dry powder formulations prepared by Examples 19, 20, and 21]

[0222] The solubility properties of the DPI formulations according to Examples 19, 20, and 21, and the comparative budesonide powder (i.e., micronized budesonide), were demonstrated by applying the adapted method described by Pilcer et al. (J Pharm Sci 2013). The comparative micronized budesonide powder was prepared by spray-drying a 3% w / v budesonide isopropanol solution (Mini-Spray Dryer B-290, Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use.

[0223] A dissolution system (Copley Scientific, Nottingham, UK) specifically developed for investigating the DPI release profile was used, adapted from the "Paddle over Disc" method (Eur. Ph. 7). To investigate the release profile of particles deposited in the lungs, fractionation of the budesonide formulation was first performed using NGI. The Stage 2 cup was selected to have a removable disc insertion section for collecting particles. Of particular interest, at the selected inhalation rate (100 L / min for 2.4 seconds), the Stage 2 cutoff diameter ranged from 6.12 to 3.42 μm, allowing for the selection of lung-targeting particles from capsules filled with appropriate amounts of micronized budesonide, as well as formulations according to Examples 20, 21, and 22, resulting in the deposition of approximately 500 μg of budesonide in Stage 3. Next, the disc insertion area was covered with a polycarbonate membrane (pore size 0.4 μm) (Merck Millipore) and placed in a paddle dissolving apparatus (Erweka DT6; ERWEKA GmbH, Heusenstamm, Hesse, Germany) filled with 400 mL of 0.01 mM phosphate buffered saline (pH 7.4).

[0224] The dissolution test was performed under sink conditions of 37±0.2℃ and pH 7.35±0.05. The paddle, set to 25±2mm between the center of the blade and the center of the disk assembly, was set to a rotation speed of 50±4rpm. A sample volume of 2.0 mL was passed through a cellulose acetate syringe filter (VWR, Leuven, Belgium) with a pore size of 0.22 mm, filtered for a preset time between 2 minutes and 24 hours, and replaced with 2.0 mL of free preheated PBS.

[0225] The 100% budesonide dissolution value corresponds to the mass deposited in Stage 3.

[0226] Figure 9 shows the release profile of budesonide from the breathable fraction of a comparative example of micronized budesonide, as well as the DPI formulations prepared by Examples 19, 20, and 21 (n=1), illustrating the controlled release profile of budesonide from the composition of the present invention and demonstrating the possibility of modifying the release profile by adjusting the drug / lipid ratio.

[0227] [Example 24. Preparation of a dried paclitaxel powder formulation for inhalation, n°1]

[0228] 0.1500% w / v paclitaxel, 2.8215% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany), and 0.0285% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm ethanol (50°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and drying air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0229] [Example 25. Preparation of a dried paclitaxel powder formulation for inhalation, n°2]

[0230] 0.600% w / v paclitaxel, 2.376% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany), and 0.024% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm ethanol (50°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0231] [Example 26. Preparation of a dried paclitaxel powder formulation for inhalation, n°3]

[0232] 1.500% w / v paclitaxel, 1.485% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany), and 0.015% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm ethanol (50°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and drying air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0233] [Example 27. Preparation of a dried paclitaxel powder formulation for inhalation, n°4]

[0234] 2.700% w / v paclitaxel, 0.297% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany), and 0.003% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm ethanol (50°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and dry air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0235] [Example 28. Preparation of a dried paclitaxel powder formulation for inhalation, n°5]

[0236] 1.50% w / v paclitaxel, 1.35% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany), and 0.15% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm ethanol (50°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and drying air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0237] [Example 29. Preparation of a dried paclitaxel powder formulation for inhalation, n°6]

[0238] 1.5% w / v paclitaxel, 1.2% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany), and 0.3% w / v TPGS (Sigma-Aldrich, St-Louis, USA) were solubilized in warm ethanol (50°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and drying air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0239] [Example 30. Preparation of a dried paclitaxel powder formulation for inhalation, n°7]

[0240] 1,500% w / v paclitaxel, 1,485% w / v hydrogenated castor oil (BASF, Ludwigshafen, Germany), 0.015% w / v TPGS (Sigma-Aldrich, St-Louis, USA), and 0.3% w / v L-leucine (Sigma-Aldrich) were solubilized in warm ethanol (50°C) under magnetic stirring, and spray-dried using a Mini-Spray Dryer B-290 (Buchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used during spray drying were as follows: feed rate 3.0 g / min, inlet temperature 70°C, 0.7 mm nozzle, 1.5 mm nozzle cap, compressed air 800 L / min, and drying air flow 35 m 3 / h. The apparatus was equipped with a B-296 dehumidifier (Buchi Labortechnik AG) and maintained the relative humidity at 50%HR during spray drying.

[0241] [Example 31. Preparation of a budesonide dry powder formulation for inhalation, n°7]

[0242] Budesonide dry powder formulation n°4 for inhalation was first prepared according to Example 19. Then, 10 g of budesonide dry powder formulation n°4 for inhalation and 30 g of lactose (Respitose® SV003, DFE Pharma, Goch, Germany) were blended homogenously in a Turbula® mixer (Willy A. Bachofen AG, Muttenz, Switzerland) to obtain a carrier-based dry powder formulation for inhalation.

[0243] It should be understood that the present invention is not limited to the embodiments described and that modifications can be applied without departing from the scope of the appended claims. [Brief explanation of the drawing]

[0244] [Figure 1]Figure 1 shows the FPF values ​​(%) of the comparative example (composition F5 in Levet et al.) and Examples 1 and 2 (mean ± standard deviation, n=2-3). [Figure 2] Figure 2 shows the cisplatin release profiles from the breathable fractions of the DPI formulations prepared in Examples 1 and 2, compared to a comparative formulation consisting solely of cisplatin microparticles. [Figure 3] Figure 3 shows the FPF value of Example 10 compared to a comparative formulation consisting only of pemetrexed microparticles (mean ± standard deviation, n=3 and 1, respectively), demonstrating the superiority of Example 10 over conventional DPI formulations in terms of pulmonary deposition. [Figure 4] Figure 4 shows the release profile of pemetrexed from the breathable fraction of the DPI formulation prepared in Example 10 compared to a comparative formulation consisting only of pemetrexed microparticles (mean ± standard deviation, n=3). [Figure 5] Figure 5 shows the FPF values ​​(mean ± standard deviation) for Example 13, demonstrating the high pulmonary deposition rate of the insulin-based DPI composition disclosed in the present invention. [Figure 6] Figure 6 shows the insulin release profile from the breathable fraction of the DPI preparation prepared in Example 13 (up to 240 minutes, mean ± standard deviation, n=2) (A) compared with the comparative preparation (up to 180 minutes) (B) described by Depreter et al. [Figure 7] Figure 7 shows the cisplatin release profile from the breathable fraction of the DPI formulation prepared in Example 16 compared to a comparative formulation consisting of cisplatin microparticles (mean ± standard deviation, n=3). [Figure 8] Figure 8 shows the FPF values ​​of Examples 19 and 20 compared to Comparative Examples BUD-TS4 and BUD-TS5 (mean ± standard deviation, n=3). (***) p<0.001, t-test. The superiority of the budesonide composition disclosed in this invention is demonstrated with respect to lung deposition compared to other triglyceride-based budesonide DPI formulations. [Figure 9]Figure 9 shows the release profile of budesonide from the breathable fraction of a comparative example of micronized budesonide, as well as the DPI formulations prepared by Examples 19, 20, and 21 (n=1), illustrating the controlled release profile of budesonide from the composition of the present invention and demonstrating the possibility of modifying the release profile by adjusting the drug / lipid ratio.

Claims

1. A dry powder inhalation preparation, At least one active pharmaceutical ingredient (API), A lipid matrix comprising at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin, and mixtures thereof, wherein the lipid matrix is ​​hydrogenated castor oil. The ratio between the at least one API and the hydrogenated castor oil is 15 / 85 to 85 / 15. The aforementioned formulation has a geometric particle size distribution (PSD) of 5 μm or less. ５０ , Geometric particle size distribution (PSD) d below 7 μm ９０ A dry powder inhalation preparation having a volume average diameter D[4,3] of 6 μm or less.

2. The dry powder inhalation formulation according to claim 1, wherein the weight ratio between the at least one API and the at least one triglyceride, API / hydrogenated castor oil, is 25 / 75 to 75 / 25 with respect to the total weight of the dry powder inhalation formulation.

3. The dry powder inhalation preparation according to claim 1, wherein the weight ratio between at least one API and hydrogenated castor oil is 40 / 60 to 60 / 40 with respect to the total weight of the dry powder inhalation preparation.

4. The dry powder inhalation formulation according to claim 1, further comprising a long-lasting pulmonary retention excipient selected from the group consisting of PEG-modified excipients or polysaccharides.

5. The dry powder inhalation preparation according to claim 4, wherein the long-term lung retention excipient is a PEG-modified excipient and is present in an amount of 0.1 wt% to 20 wt% of the total weight of the dry powder inhalation preparation.

6. The dry powder inhalation preparation according to claim 1, further comprising an excipient in an amount of 0.1 wt% to 80 wt% with respect to the total weight of the dry powder inhalation preparation, wherein the excipient is selected from the group consisting of sugar alcohols; polyols; crystalline sugars; inorganic salts; organic salts; oligosaccharides; titanium dioxide; silicone dioxide; magnesium stearate; lecithin; amino acids; derivatives of amino acids such as acesulfame K or aspartame; lauric acid or its esters and salts; palmitic acid or its esters and salts; stearic acid or its esters and salts; erucic acid or its esters and salts; behenic acid or its esters and salts; sodium stearyl fumarate; sodium stearyl lactate; phosphatidylcholine; phosphatidylglycerol; natural and synthetic pulmonary surfactants; triglycerides; sugar esters; phospholipids; cholesterol; and talc.

7. The aforementioned APIs include budesonide, salbutamol, fluticasone, beclomethasone, mometasone, ciclesonide, formoterol, salbutamol, alformoterol, indacaterol, orodaterol, salmeterol, ipratropium, acridinium, glycopyrronium, tiotropium, unmeclidinium, mometasone, ciclesonide, formoterol, alformoterol, ibuprofen, tobramycin, vancomycin, tetrahydrolipstatin, clarithromycin, isoniazid, and rifa. Pyraminamide, pyrazinamide, itraconazole, voriconazole, aztreonam, ethambutol, streptomycin, kanamycin, amikacin, colistin, colistimethasoderm, capreomycin, ciprofloxacin, rifapentin, doxycycline, cycloserine E, ethionamide, gatifloxacin, levofloxacin, moxifloxacin, ofloxacin, fosfomycin, p-aminosalicylic acid, denufosol tetrasodium, lancobutide, ribavirin, zanamivir, laminavir, rupintrivir A small chemical molecule having bronchodilation activity, glucocorticoid activity, anti-inflammatory activity, or anti-infective activity, which is adsorbed to the lungs for systemic or topical treatment, selected from the group consisting of pentamidine, amphotericin B, posaconazole, isabconazole, capsufungin, micafungin, anidurafungin, iloprost, levothyroxine, their salts, solvates, hydrates, and esters, and combinations thereof, or the API consists of peptides, proteins, antibodies, antibody fragments, nanobodies, and nucleic acids. The API is a macromolecule selected from the group, and the APIs include insulin, proinsulin, synthetic insulin, semi-synthetic insulin, bevacizumab, pembrolizumab, atezolizumab, nivolumab, ipilimumab, Toll-like receptor agonists, ghrelin, IgG monoclonal antibodies, small interfering ribonucleic acid (siRNA), dorunase alpha, cyclosporine A, alpha-1 antitrypsin, interleukin antagonists, interferon-α, interferon-β, interferon-γ, interferon-ω, interleukin-2, and anti-IgEThe dry powder inhalation formulation according to claim 1, wherein the API is selected from the group consisting of mAb, catalase, calcitonin, paratyloid hormone, human growth hormone, insulin-like growth factor-I, heparin, rhG-CSF, GM-CSF, Epo-Fc, FSH-Fc, sFc-γRIIb, and mRNA, or the API is an antitumor agent.

8. The dry powder inhalation formulation according to claim 7, wherein the API is an antitumor agent selected from the group consisting of cisplatin, carboplatin, oxaliplatin, docetaxel, paclitaxel, pemetrexed, etoposide, and vinorelbine.

9. The dry powder inhalation formulation according to claim 8, wherein the API is cisplatin, and the ratio between the at least one API and hydrogenated castor oil is 40 / 60 to 60 / 40.

10. A dried powder inhalation preparation according to claim 9, comprising a disaccharide excipient.

Images