RU2817158C2 - Dry powder inhalation composition and its use for pulmonary drug treatment - Google Patents

Abstract

FIELD: medicine; pharmaceutics.

SUBSTANCE: dry powder inhalation composition containing at least one API and a lipid matrix containing at least one triglyceride, selected from a group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin and a mixture thereof, and a method for preparing it.

EFFECT: dry powder inhalation composition and its use for pulmonary drug treatment are proposed.

23 cl, 9 dwg, 31 ex

Description

The present invention relates to a dry powder inhalation composition and its use for drug treatment of the lungs.

Dry powder inhalation (DPI) therapy is now widely used for many types of pulmonary treatments, such as asthma, and much effort has been devoted to optimizing dry powder inhalation formulations to prolong the retention of the active pharmaceutical ingredient (API) in the lungs.

Inhalation allows high doses of a drug to be administered directly into the lungs without prior distribution in the body. This allows, on the one hand, to use a smaller amount of active pharmaceutical ingredient (API), in addition, it is possible to reduce its loss in the body, but also, in the case of probable API toxicity, direct in situ administration can reduce toxicity.

Moreover, inhalation is a promising route of administration for delivering APIs into the systemic circulation because the lungs have a huge absorption surface.

However, traditional immediate-release dry powder inhalation (DPI) formulations may remain in the lungs for too short a time due to multiple mechanisms of clearance of exogenous inhaled particles (i.e., mucociliary and macrophage clearance) and rapid dissolution of drug particles, which leads to rapid absorption into the systemic circulation. Consequently, such short retention times require multiple doses, resulting in poor patient compliance. Immediate release of API may also result in poor local tolerability due to rapid dissolution of drug particles and high peak concentrations of API in pulmonary secretions following pulmonary deposition.

Certain excipients have been found to prolong pulmonary residence time to achieve a sustained pulmonary residence profile of the API.

Thus, there is a need to develop additional dry powder inhalation formulations to improve the effectiveness of pulmonary therapy.

In this context, for example, controlled-release cisplatin-based DPI formulations with high drug content and high fine particle fraction have been developed (Levet et al, Int J Pharm 2016). The formulations have demonstrated controlled release and long-lasting properties in the lungs, resulting in low systemic distribution (Levet et al, Int J Pharm 2017).

In this regard, Levet et al. developed a specific formulation for one antitumor agent, namely cisplatin-containing pegylated excipients, to prevent too rapid elimination through a pulmonary epithelium-based protective mechanism, and also compared different carriers such as polymers and lipid matrix to provide a long residence time in the lungs with increased thereby the effectiveness of inhalation.

Among other things, the lipid matrix described by Levet et al. contains tristearin, which has been found to be a promising potential lipid matrix agent.

In addition, although a lipid-derived matrix-forming excipient is preferred over polymers, due to toxicity, only a few substances capable of forming a lipid matrix have been cleared and approved for use in an inhaled pharmaceutical composition compared to other drug treatments (Pilcer et Amighi, Int J Pharm 2010).

Unfortunately, although tristearin is a promising potential agent, its availability on the market as a pharmaceutical compound is quite limited.

The present invention seeks to overcome at least a portion of these disadvantages by providing a dry powder inhalation composition, for example for use in monotherapy or polytherapy, containing at least one active pharmaceutical ingredient (API) and a lipid matrix containing at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin and mixtures thereof.

Surprisingly, it has been discovered that said at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin and a mixture thereof, for which a pharmaceutical grade of purity exists, is capable of forming a lipid matrix with the active pharmaceutical ingredient (API) and exhibiting increased deposition rates in lungs, which is suitable for use by inhalation into the lungs.

Indeed, monohydroxystearin, dihydroxystearin, trihydroxystearin and mixtures thereof are compounds having physicochemical properties such as Log P values above 5, preferably in the range of 14 to 25, more particularly from 18 to 25 and preferably about 20, when measured by shaking in the flask, and/or a melting point that is greater than or equal to 40°C, preferably greater than or equal to 60°C, more preferably greater than or equal to 75°C.

This allows the inhalation composition to be provided in the form of a dry powder, rather than an oil phase or sticky paste, for proper use in a dry powder inhaler.

In addition, in accordance with the present invention, it has surprisingly been found that it is possible to increase the rate of deposition of API in the lungs relative to other conventional triglycerides, such as tristearin, and provide a modulated dissolution profile of the API by changing the ratio between the amount of API and the amount of at least one triglyceride, selected from the group containing monohydroxystearin, dihydroxystearin, trihydroxystearin and a mixture thereof, which is not typical for traditional triglycerides.

Preferably, the dry powder inhalation composition in accordance with the present invention has a weight ratio between said at least one API and said at least one triglyceride: API/triglyceride in the range of 0.1/99.9 to 99.9/0.1, preferably from 10/90 to 88/12, preferably from 15/85 to 85/15, preferably from 25/75 to 75/25, more preferably from 30/70 to 70/30, in particular from 40/60 to 60/ 40, for example, is about 50/50.

Advantageously, in the composition of the present invention, said at least one triglyceride is hydrogenated castor oil.

Castor oil is described in the literature in long lists of standard excipients (see US 2005/0042178) or grinding agents (WO2013/128283).

In a preferred embodiment in accordance with the present invention, the dry powder inhalation composition contains from 0.1 wt%. up to 98% wt. of said at least one API relative to the total weight of the dry powder inhalation composition. In a first specific embodiment of the present invention, said active agent is a small chemical molecule having a solubility in alcohols of at least 0.1% w/v. or at least 0.5% w/v, more preferably at least 1% w/v, more particularly at least 5% w/v. According to this first specific embodiment, said small chemical molecule is any active pharmaceutical ingredient (API) having a solubility in alcohols of 0.1% w/v. or more, defined in the present invention as an alcohol-soluble API, such as, 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, glycopyronium bromide, tiotropium bromide, umeclidinium bromide, ibuprofen, vancomycin, vancomycin hydrochloride, tetrahydrolipstatin, isoniazid, rifampicin, pyrazinamide, docetaxel, vincristine, vincristine sulf at, etoposide, vinorelbine, gemcitabine and etc.

More specifically, the dry powder inhalation composition in accordance with the first specific embodiment contains from 0.1 wt%. up to 95% wt., preferably from 1% wt. up to 90% wt., more preferably from 1% wt. up to 85% wt. of said at least one API relative to the weight of the dry powder inhalation composition.

In a preferred embodiment in accordance with the present invention, the dry powder inhalation composition consists of lipid matrix particles in which the alcohol-soluble API is dissolved or finely dispersed.

In a second specific embodiment of the present invention, said active agent is a small chemical molecule having a solubility in alcohols of less than 0.5% w/v. In this second specific embodiment, said small chemical molecule is any API having a solubility in alcohols of less than 0.5% w/v, in particular less than 0.1% w/v, and in particular less than 0.01 % w/v, defined as an alcohol-insoluble API such as, for example, cisplatin, carboplatin, oxaliplatin, pemetrexed disodium, azacitidine, beclomethasone dipropionate, tobramycin, aclidinium bromide, etc.

More specifically, the dry powder inhalation composition in accordance with the second specific embodiment contains from 0.1 wt%. up to 95% wt., preferably from 1% wt. up to 90% wt., more preferably from 1% wt. up to 85% wt. of said at least one API relative to the weight of the dry powder inhalation composition.

Accordingly, the dry powder inhalation composition contains from 0.1% wt. up to 95% wt. the specified alcohol-insoluble API, preferably from 1% of the mass. up to 90% wt., more preferably from 1% wt. up to 85% wt., for example from 1% wt. up to 60 or from 1 to 50 wt.%, relative to the total weight of the specified dry powder inhalation composition.

In a preferred embodiment in accordance with the present invention, the dry powder inhalation composition consists of lipid matrix particles in which the alcohol-insoluble API is dispersed.

In a third specific embodiment according to the present invention, said active agent is a macromolecule. In this third specific embodiment, in accordance with the present invention, the dry powder inhalation composition contains from 0.1 wt%. up to 95% wt., preferably from 1% wt. up to 90% wt., more preferably from 1% wt. up to 85% wt. of said at least one API relative to the weight of the dry powder inhalation composition.

Accordingly, the dry powder inhalation composition contains from 0.1% wt. up to 95% wt. the specified macromolecule, preferably from 1% of the mass. up to 90% wt., more preferably from 1% wt. up to 85% wt., for example, from 1% wt. up to 60 or from 1 to 50 wt.%, relative to the total weight of the specified dry powder inhalation composition.

In yet another preferred embodiment, the composition in accordance with the present invention further contains a long-lasting excipient in the lungs, such as a PEGylated excipient, or polysaccharides, such as chitosan or dextran.

In accordance with the present invention, said long-lasting excipient in the lungs is preferably a PEGylated excipient and is present in an amount ranging from 0.1% by weight. up to 20% by weight, preferably from 0.2 to 10% by weight, more preferably from 0.5 to 5% by weight. relative to the total weight of said dry powder inhalation composition.

More specifically, in accordance with the present invention, said PEGylated excipient is derived from vitamin E or from phospholipids such as tocopherol polyethylene glycol succinate (TPGS) or distearoylphosphoethanolamine polyethylene glycol 2000 (DSPE-mPEG-2000).

In yet another preferred embodiment, the composition in accordance with the present invention further contains one or more excipients, such as excipients that improve the physicochemical and/or aerodynamic properties of the dry powder for inhalation.

By "one or more excipients" is meant a compound selected from sugar alcohols; polyhydric alcohols (such as sorbitol, mannitol and xylitol); crystalline sugars, including monosaccharides (such as glucose, arabinose) and disaccharides (such as lactose, maltose, sucrose, dextrose, trehalose, maltitol); inorganic salts (such as sodium chloride and calcium carbonate); organic salts (such as sodium lactate, potassium or sodium phosphate, sodium citrate, urea); polysaccharides (such as dextran, chitosan, starch, cellulose, hyaluronic acid and their derivatives); oligosaccharides (such as cyclodextrins and dextrins); titanium dioxide; silicon dioxide; magnesium stearate; lecithin; amino acids (such as leucine, isoleucine, histidine, threonine, lysine, valine, methionine, phenylalanine); amino acid derivatives (such as acesulfame K, aspartame); lauric acid or its derivatives (such as esters and salts); palmitic acid or its derivatives (such as esters and salts); stearic acid or derivatives thereof (such as esters and salts); erucic acid or its derivatives (such as esters and salts); behenic acid or its derivatives (such as esters and salts); sodium stearyl fumarate; sodium stearyl lactylate; phosphatidylcholines; phosphatidylglycerols; natural and synthetic light surfactants; lauric acid and its salts (such as sodium lauryl sulfate, magnesium lauryl sulfate); triglycerides; sugar esters; phospholipids; cholesterol; talc.

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

In certain preferred embodiments, the excipient is phospholipids or cholesterol.

In certain preferred embodiments, the excipient is mannitol, dextran, hyaluronic acid, lactose, phospholipids or cholesterol.

In certain preferred embodiments, the excipient is mannitol, dextran, phospholipids or cholesterol.

In certain embodiments, said at least one excipient is a carrier.

In one preferred embodiment of the composition in accordance with the present invention, the composition is in the form of fine particles having a geometric particle size distribution (PSD) d ₅₀ that is less than or equal to 30 μm, preferably less than 15 μm, preferably less than or equal to 10 μm, preferably less or equal to 5 µm.

In a further preferred embodiment of the composition in accordance with the present invention, said composition is in the form of fine particles having a geometric particle size distribution (PSD) d ₉₀ that is less than or equal to 60 μm, preferably less than or equal to 30 μm, preferably less than or equal to 15 μm, preferably less than or equal to 10 µm, and more preferably less than or equal to 7 µm.

In yet another further preferred embodiment of the composition in accordance with the present invention, said composition is in the form of particles having an average volumetric diameter D[4,3] that is less than or equal to 40 μm, preferably less than or equal to 20 μm, preferably less than or equal to 15 µm, preferably less than or equal to 10 µm, preferably less than or equal to 6 µm.

Preferably, in accordance with the present invention, said improved excipient and/or said excipient is present in an amount ranging from 0.1% w/w. up to 80% by weight, preferably less than 70% by weight, more preferably less than 60% by weight, in particular less than 50% by weight, for example less than 50% by weight, of the total weight of said dry powder inhalation composition.

The term "fine particle dose" or FPD essentially refers to the mass of particles with an aerodynamic diameter less than 5 μm relative to the mass of the nominal dose (ie, the mass of the dose delivered to the inhalation device).

The fine particle dose or fine particle fraction is the fraction of a pharmaceutical formulation that can be deeply inhaled and which theoretically may exhibit pharmacological activity (Dunbar et al, Kona 16: 7–45, 1998).

In an advantageous embodiment according to the present invention, the composition is in the form of fine particles having a mass median aerodynamic diameter (MMAD) that is less than or equal to 6 μm, preferably less than or equal to 5 μm, preferably less than or equal to 4 μm.

MMAD refers to the diameter of the particles deposited in the impactor, with 50% (w/w) of the particles having a smaller diameter and 50% (w/w) having a larger diameter.

The terms "aerodynamic diameter" or dae of a particle can be defined as the diameter of a sphere of unit density (i.e. density equal to 1) that has the same settling rate in still air as the particle in question. The dae parameter effectively measures respirable particles and takes into account factors affecting their aerodynamic properties. The parameter dae can be used to compare particles of different physical sizes, taking into account their density and shape, as well as their geometric size.

Methods for measuring dae are those described in the European Pharmacopoeia or the United States Pharmacopoeia using an impactor or impactor device, such as a glass impinger, a multistage liquid impinger (MsLI), a cascade impactor, or a next generation impactor (NGI). This allows the aerodynamic properties of DPI formulations (including MMAD, geometric standard deviation, lung deposition type, fine particle dose, fine particle fraction) to be measured under simulated breathing conditions.

The total dose of particles with aerodynamic diameters less than 5 µm can be calculated by interpolation from the data collection efficiency curve and taken as the fine particle dose (FPD) or fine particle fraction (FPF), expressed as a percentage of the nominal dose of the API (i.e. dose contained in the DPI device).

Preferably, the dry powder inhalation composition in accordance with the present invention is packaged, for example, in a blister or capsule for use in a dry powder inhaler or in a sealed and/or disposable dry powder inhaler.

In a specific embodiment, said active agent is a small chemical molecule having bronchodilator activity, glucocorticoid activity, anti-inflammatory activity, anti-infective activity (eg, antibiotics, antituberculosis agents, antifungals, antivirals), etc.

In another specific embodiment, said small chemical molecule is any pulmonaryly absorbable active pharmaceutical ingredient for systemic or topical therapy, such as, for example, budesonide, salbutamol, fluticasone, beclomethasone, mometasone, ciclesonide, formoterol, salbutamol, arformoterol, indacaterol, olodaterol, salmeterol, ipratropium, aclidinium, glycopyrronium, tiotropium, unmeclidinium, mometasone, ciclesonide, formoterol, arformoterol, ibuprofen, tobramycin, vancomycin, tetrahydrolipstatin, clarithromycin, isoniazid, rifampicin, pyrazinamide, itraconazole, voriconazole, m, ethambutol, streptomycin, kanamycin, amikacin, colistin, colistimethate sodium, capreomycin, ciprofloxacin, rifapentine, doxycycline, cycloserine E, ethionamide, gatifloxacin, levofloxacin, moxifloxacin, ofloxacin, fosfomycin, p-aminosalicylate, denufozole tetrasodium, lancovutide, ribavirin, zanamivir, rupintrivir, pentamidine, amphotericin B, posaconazole, isavuconazole, capsufungin, micafungin, anidulafangin, iloprost, levothyroxine, their salts, solvates, hydrates, polymorphic forms and their esters, their combinations, analogs and derivatives.

In yet another embodiment, said active agent is a macromolecule such as a peptide, protein, antibody, antibody fragment, nanobody, nucleic acid.

Preferably, the macromolecule specified in accordance with the present invention is insulin, proinsulin, synthetic insulin, semi-synthetic insulin, bevacizumab, pembrolizumab, atezolizumab, nivolumab, ipilimumab, toll-like receptor agonists, ghrelin, IgG monoclonal antibody, small interfering ribonucleic acid (siRNA), dornase alpha, cyclosporine A, antitrypsin alpha-1, interleukin antagonists, interferon-α, interferon-β, interferon-γ, interferon-ω, interleukin-2, monoclonal antibody to IgE, catalase, calcitonin, parathyroid hormone, human growth hormone, insulin-like growth factor I, heparin, rhG-CSF, GM-CSF, Epo-Fc, FSH-Fc, sFc-γ RIIb, mRNA.

In a further preferred embodiment, said active agent is an antitumor agent, which is, for example, used in lung cancer and lung tumors.

Lung cancer is the cancer with the highest prevalence and mortality rate in the world. In most cases, lung cancer is diagnosed in advanced stages. Consequently, patients often already have metastases in the lungs or other organs, e.g. extrapulmonary metastases. Treatment modalities are mainly used in combination and include surgery, radiotherapy, chemotherapy, targeted therapy and immunotherapy.

Up to 60% of patients with lung cancer undergo chemotherapy, mostly in advanced stages of the disease. Chemotherapy is currently administered by intravenous injection or infusion or per os, i.e. a systemic route of administration is used, i.e. systemic chemotherapy. Chemotherapy is associated with severe systemic toxicity due to (i) the widespread distribution of the chemotherapy drug in the body and (ii) lack of selectivity for cancer cells. Consequently, oncologists are in great need of new, more effective and better tolerated treatments.

In reality, chemotherapy is the search for the greatest amount of anticancer agent to reduce as much as possible the irreversible toxicity-related side effects and unpleasant side effects caused by chemotherapy.

Accordingly, a very complex therapeutic regimen is expected, combining very often different therapeutic modalities and actions, such as, for example, surgery, with a first cycle of radiotherapy, followed by complex cycles of injections or intravenous infusion of an antitumor agent, which may be the same or different in each cycle and can decrease or adapt depending on the side effect detected on the patient's body.

Each antitumor agent or molecule has dose-limiting toxicity (DLT), such as nephrotoxicity, neurotoxicity, etc., which requires the introduction of a rest period(s) into the therapeutic regimen, allowing the patient's body to recover from unwanted side effects if they are reversible.

In addition, the antitumor agent has a half-life, which is often referred to as its half-life. The half-life is the period of time required for the concentration or amount of a drug in the body to be reduced by half. The half-life of an antineoplastic agent is typically between 12 hours and 36 hours, and may be short because it corresponds to the time of beneficial effects of the antineoplastic agent on the human body.

However, due to the dose-limiting toxicity (DLT) of the antineoplastic agent, the number of doses administered over multiple dosing periods remains limited and must be separated from each other, as previously stated, by a period of rest or withdrawal.

The combination of factors such as half-life, dose-limiting toxicity, and administration of the antineoplastic agent orally or by injection or intravenous infusion results in the concentration of antineoplastic agent that effectively reaches the solid tumor site being low and having limited effect on the tumor itself and leads to high systemic toxicity for the patient.

Another limitation of an antineoplastic agent is the cumulative dose, which is the total dose resulting from repeated exposure of the same part of the body or the entire body to the antineoplastic agent.

For cisplatin administered intravenously, the cumulative dose is 300 mg/m2 over 4 to 6 cycles.

These facts have enabled the present inventors to develop more targeted treatments and site-of-treatment injection/infusion.

There are different types of inhalation therapy, and they use nebulizers, inhalers that deliver APIs into the airways in different ways. Inhalers can also be of different types, and one of them is dry powder inhalers (DPI).

Compared to nebulizers, dry powder inhalers (DPIs) are well suited for chemotherapy. They allow the administration of high doses of APIs as well as poorly water-soluble compounds (i.e., most chemotherapeutic agents in the treatment of cancer). Moreover, DPI reduces environmental aerosol contamination due to (i) only their activation and initiation by the patient's inhaled flow and (ii) the negligible amount of exhaled drug doses. Finally, the DPI can be designed as a single-use device.

Thus, there is a need to develop a pulmonary delivery method for an anticancer agent and to deeply adapt existing inhalation devices and formulations used in clinical trials to deliver effective anticancer therapeutics to tumor-affected lungs.

There are many advantages to prolonging the residence profile of the antineoplastic agent in the lungs and not being absorbed too quickly. However, achieving a sustained release profile currently remains problematic because the pulmonary absorption surface area is very large, resulting in rapid systemic absorption of the agent.

As previously explained, controlled-release cisplatin DPI formulations have been developed with high drug content and a high fraction of fine particles, which are characterized by controlled release and long residence in the lungs, resulting in low systemic distribution (Levet et al , Int J Pharm 2017). As reported herein, in a mouse model of lung cancer, this approach resulted in comparable tumor responses in vivo compared to a half-dose intravenous regimen (1.0 mg/kg and 0.5 mg/kg, respectively). The intravenous regimen shows good results for extrapulmonary metastases, while inhaled cisplatin appears to be more active against lung tumors.

DPI formulations consisting of paclitaxel-based nanocarriers with extended lung residence time, limited systemic distribution, and specifically targeting lung cancer cells by targeting the folate receptor (FR) have also been developed (Rosière et al, Int J Pharm 2016; Rosière et al, Mol Pharm, 2018). FR, especially FR-α, is overexpressed on cancer cell surfaces in many lung tumors (ie, more than 70% of adenocarcinomas) and is a promising membrane receptor for targeting in lung cancer. In a mouse model of lung cancer, significantly higher survival rates were observed with FR-targeted inhaled treatment in combination with intravenous Taxol® (a commercial form of paclitaxel) than with Taxol® alone.

It can be concluded that although the use of DPI chemotherapy has led to confirmation of preclinical results in terms of pharmacokinetic profiles and safety, its effectiveness is still quite limited.

Although achieving a sustained release profile of an antineoplastic agent can have many benefits, in the context of inhaled chemotherapy this issue has also received attention because the lungs have a very large absorption surface area, resulting in the API being rapidly absorbed and released into the systemic circulation.

In addition, since the lungs are equipped with strong protective mechanisms, elimination of APIs during inhalation therapy occurs quite quickly due to the highly efficient clearance mechanisms in the lungs for foreign deposited particles.

Sebti et al (Sebti et al, Eur J Pharm Biopharm 2006a; Sebti et al, Eur J Pharm Biopharm 2006b) developed solid lipid particulates with budesonide and a cholesterol lipid matrix, but did not observe any delay in the release profile of budesonide.

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

For the same purpose as above, Levet et al. were able to prolong residence time in the lungs, thereby increasing the effectiveness of inhaled chemotherapy, and identified the lipid matrix of tristearin and TPGS as a promising potential agent.

Additionally, although a lipid-derived matrix-forming excipient is preferred over polymers, due to toxicity, only a few substances capable of forming a lipid matrix are permitted and approved for use in an inhaled pharmaceutical composition compared to other drug treatments (Pilcer and Amighi, Int J Pharm 2010).

Unfortunately, although the tristearin matrix has been identified as promising, its availability on the market as a pharmaceutical compound is quite limited, while the availability of the at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin and mixtures thereof, significantly higher.

Moreover, the tristearin-based matrix was not promising for controlling the release profile of voriconazole, whereas the preparation method (ie, spray drying) and excipient composition were similar to those described by Levet et al in the context of cisplatin.

In addition, in accordance with the present invention, it has been found that said at least one triglyceride selected from the limited group in accordance with the present invention allows, in addition to obtaining a slow release profile and prolonged residence of the chemotherapeutic agent in the lungs, to significantly increase the rate of deposition lungs compared to the closest triglyceride tristearin.

Preferably, said antitumor agent is cisplatin, carboplatin, oxaliplatin, docetaxel, paclitaxel, pemetrexed, etoposide, vinorelbine.

Other embodiments of dry powder inhalation compositions in accordance with the present invention are mentioned in the accompanying claims.

The present invention relates to methods for producing a dry powder inhalation composition in accordance with the present invention, comprising the following steps:

a) mixing one or more APIs with a predetermined amount of at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin and a mixture thereof, with or without a solvent;

b) an ascending or descending method resulting in the production of respirable particles from the previous mixture, such as a fine grinding step such as spray drying of a suspension or solution, spray crystallization of a solution of the active drug or API in said at least one triglyceride, or extrusion followed by grinding in a jet mill, a physical mixture of the API with said at least one triglyceride.

For example, the present invention relates to a method of preparing a dry powder inhalation composition in accordance with the present invention, comprising the step of suspending or dissolving the powder of one or more APIs in a predetermined amount of at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin and mixture to form a particle suspension or solution of one or more APIs (e.g., by melting and/or extrusion) followed by reduction in particle size of the solution or suspension of one or more APIs obtained after cooling (e.g., by spray crystallization) or high speed extrusion and/or pressure to obtain said dry powder inhalation composition (eg, by grinding in a jet mill).

In another example, the manufacturing method includes the following steps:

a) homogeneously mixing at least one API with at least one triglyceride to form a homogeneous mixture,

b) extruding the homogeneous mixture using a (twin-screw) extruder at an appropriate temperature to obtain a homogeneous lipid matrix containing said API,

c) cutting the extrudate to produce large granules/cylinders,

d) optionally converting the triglycerides into a stable polymorphic form by storing said large granules/cylinders under appropriate storage conditions, and

e) grinding the granules using a suitable mill to obtain microparticles for inhalation (DPI) containing said API with said at least one triglyceride.

In yet another example, the present invention relates to a method for producing a dry powder inhalation composition in accordance with the present invention, comprising the following steps:

a) suspending or dissolving a powder of one or more APIs in a solvent to form a suspension of particles or a solution of one or more APIs,

b) optionally reducing the size of said particles of one or more APIs at high speed and/or pressure of homogenization with cooling to form a suspension of particles of one or more reduced size APIs,

c) mixing a predetermined amount of at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin and a mixture thereof, in a solvent with said microcrystalline suspension or solution to obtain a mixture of said at least one API with said at least one triglyceride ,

d) spray drying a mixture of said at least one API with said at least one triglyceride to produce said dry powder inhalation composition.

In a preferred embodiment of the method according to the present invention, said high speed applied to reduce particle size is from 10,000 to 30,000 rpm, preferably from 15,000 to 26,000 rpm, and is applied over a period of time 8 to 15 minutes, preferably 9 to 12 minutes.

Preferably, in accordance with the present invention, said high pressure in the homogenization step is gradually increased from a first pressure ranging from 2000 to 10,000 psi. inch, preferably 4000 to 6000 psi. inch, for a predetermined number of pre-grinding cycles ranging from 8 to 12, preferably 9 to 11, to a second pressure ranging from 8,000 to 12,000 psi. inch, preferably 9,000 to 11,000 psi. inch, for a predetermined number of pregrind cycles ranging from 8 to 12, preferably 9 to 11, to a third pressure ranging from 18,000 to 24,000 psi. inch, preferably 19,000 to 22,000 psi. inch, for a predetermined number of pregrind cycles ranging from 18 to 22, preferably 19 to 21.

In a particularly preferred embodiment according to the present invention, said microcrystalline microcrystalline suspension has a geometric particle size distribution (PSD) d ₅₀ that is less than or equal to 30 μm, preferably less than 15 μm, preferably less than or equal to 10 μm, preferably less than or equal to 5 μm.

In a further particularly preferred embodiment according to the present invention, said microcrystal of the microcrystalline suspension has a geometric particle size distribution (PSD) d ₉₀ that is less than or equal to 60 μm, preferably less than or equal to 30 μm, preferably less than or equal to 15 μm, preferably less than or equal to 10 µm, and more preferably less than or equal to 7 µm.

In another particularly preferred embodiment according to the present invention, said microcrystal of the microcrystalline suspension has an average volume diameter D[4,3] that is less than or equal to 40 μm, preferably less than or equal to 20 μm, preferably less than or equal to 15 μm, preferably less than or equal to equal to 10 μm, and more preferably less than or equal to 6 μm.

In an advantageously preferred embodiment according to the present invention, PEGylated excipients or other excipients are additionally added.

Other embodiments of the method according to the present invention are mentioned in the accompanying claims.

The present invention further relates to the use of a dry powder inhalation composition in pulmonary therapy.

Preferably, the use according to the present invention is for the treatment of local lung diseases: asthma, COPD, pulmonary infections (eg, patients with cystic fibrosis, aspergillosis, tuberculosis, etc.), or systemic diseases (eg, diabetes, pain, etc. .).

In an embodiment of the present invention, the dry powder inhalation chemotherapy formulation is used in polytherapy for the treatment of lung cancer, such as any lung tumor, such as pulmonary metastases, such as osteosarcoma metastases, small cell lung cancer, or non-small cell lung cancer.

Advantageously, said polytherapy comprises one primary therapy selected from the group consisting of intravenous injection or infusion of a chemotherapeutic agent, immunotherapy, ablative tumor surgery, ablative surgery to remove part or all of an organ affected by the tumor, curative surgery, radiotherapy, and combinations thereof. , and one or more inhaled chemotherapy drugs as adjunctive therapy.

The present invention also relates to a corresponding method of therapy.

Detailed Description of the Invention

Other characteristics and advantages of the present invention will be obtained from the following non-limiting description and with reference to examples and figures.

In fig. 1 shows the FPF values (%) in the comparative example (composition F5, Levet et al) and in examples 1 and 2 (mean ± SD, n = 2–3).

In fig. 2 shows the release profiles of cisplatin from the inhalable fraction of DPI formulations prepared in accordance with Examples 1 and 2, relative to a comparative formulation consisting of cisplatin microparticles only.

In fig. 3 presents the FPF value of Example 10 relative to a comparative microparticulate pemetrexed formulation only (mean ± SD, n = 3 and 1, respectively), demonstrating the superiority of Example 10 relative to the standard DPI formulation in terms of pulmonary deposition.

In fig. 4 shows the release profiles of pemetrexed from the inhalable fraction of the DPI formulation prepared in accordance with Example 10 relative to a comparative formulation consisting of pemetrexed microparticles only (mean ± SD, n = 3).

In fig. 5 shows the FPF values from Example 13 (mean ± SD) demonstrating the high lung deposition rate of the insulin-based DPI composition described in the present invention.

In fig. 6 shows the insulin release profiles from the inhaled fraction of DPI formulations prepared in accordance with Example 13 (up to 240 min, mean ± SD, n = 2) (A), relative to the comparative formulations described by Depreter et al (up to 180 min) (B).

In fig. 7 shows the release profiles of cisplatin from the inhaled fraction of the DPI formulation prepared in accordance with Example 16 relative to a comparative formulation consisting of microparticulate cisplatin (mean ± SD, n = 3).

In fig. 8 shows the FPF values from Examples 19 and 20 relative to Comparative Examples BUD-TS4 and BUD-TS5 (mean ± SD, n = 3). (***) p < 0.001, t-test demonstrating the superiority of the budesonide compositions described in the present invention compared to other triglyceride-based budesonide DPI formulations in terms of pulmonary deposition.

In fig. 9 shows the release profiles of budesonide from the inhaled fraction of a comparative example of micronized budesonide and DPI formulations prepared in accordance with Examples 19, 20 and 21 (n = 1), which indicate a controlled release profile of budesonide from the compositions of the present invention and the ability to modulate the release profile by modulation of the drug/lipid ratio.

Examples

Example 1. Preparation of dry powder composition of cisplatin for inhalation No. 1

Briefly, crude cisplatin microcrystals from bulk powder (Shanghai Jinhe Bio-technology Co., Ltd., Shanghai, China) were first suspended in 50 mL of isopropanol to achieve a concentration of 5% w/v. and reduced in size by homogenization at high speed (10 min at 24,000 rpm) (X620 motor and T10 dispersing shaft, Ingenieurbüro CAT M. Zipperer GmbH, Staufen, Germany) and high pressure (EmulsiFlex-C5 high-pressure homogenizer, Avestin Inc., Ottawa, Canada) at 5000 psi. inch, then at 10,000 psi. inch for 10 pre-grind cycles and then at 20,000 psi. inch for 20 grinding cycles. The heat exchanger was connected to a homogenizing valve and maintained at −15°C using an F32-MA cooling circulator (Julabo GmbH, Seelbach, Germany). At the end of the process, an aliquot was removed from the suspension to measure the particle size distribution (PSD) of the cisplatin microcrystals using laser diffraction (see below).

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

The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity (RH) of 50% RH during spray drying.

The geometric PSD of bulk cisplatin, microparticles of cisplatin produced by the size reduction method, and dry powder formulation of cisplatin were measured as suspended and individualized particles. A Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used for this purpose. Bulk cisplatin and aliquots obtained from the size reduction process were measured in cisplatin-saturated isopropanol. The dry powder formulation of cisplatin was pre-dispersed, shaken and measured in cisplatin-saturated 0.1% w/v. poloxamer 407 (BASF, Ludwigshafen, Germany) and 0.9% aqueous NaCl solution. PSD was expressed as the median volume diameter d(0.5) (when 50% of the particles are smaller than the expressed diameter), the median volume diameter D[4,3], and the percentage of fine particles determined from the cumulative reduced curves (% particles less than 5 μm). The PSD results are within the range provided in accordance with the present invention.

Example 2. Preparation of dry powder formulations of cisplatin for inhalation No. 2

Briefly, crude cisplatin microcrystals from bulk powder (Shanghai Jinhe Bio-technology Co., Ltd., Shanghai, China) were first suspended in 50 mL of isopropanol to achieve a concentration of 5% w/v. and reduced in size by homogenization at high speed (10 min at 24,000 rpm) (X620 motor and T10 dispersing shaft, Ingenieurbüro CAT M. Zipperer GmbH, Staufen, Germany) and high pressure (EmulsiFlex-C5 high-pressure homogenizer, Avestin Inc., Ottawa, Canada) at 5000 psi. inch, then at 10,000 psi. inch for 10 pre-grind cycles and then at 20,000 psi. inch for 20 grinding cycles. The heat exchanger was connected to a homogenizing valve and maintained at −15°C using an F32-MA cooling circulator (Julabo GmbH, Seelbach, Germany). At the end of the process, an aliquot was removed from the suspension to measure the particle size distribution (PSD) of the cisplatin microcrystals using laser diffraction (see below).

Hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA) dissolved in heated isopropanol were then added to the microcrystalline suspension to obtain a final cisplatin concentration of 1.0% w/v. and a mixture (99: 1 w/w) of hydrogenated castor oil/TPGS 1.0% w/v. and spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

The geometric PSD of bulk cisplatin, microparticles of cisplatin produced by the size reduction method, and dry powder formulation of cisplatin were measured as suspended and individualized particles. For this purpose, a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used as described in Example 1.

Example 3. Analysis of the deposition rate of dry powder formulations of cisplatin obtained in accordance with examples 1 and 2

The compositions were analyzed in accordance with the comparative example (composition F5, Levet al) and in accordance with examples 1 and 2 in relation to the values of the fine particle fractions.

Fine particle fraction (FPF), a percentage related to the extracted dose of cisplatin-based particles with an aerodynamic diameter (dae) of less than 5 μm and an aerodynamic PSD characterized by mass median aerodynamic diameter (MMAD), was determined using MsLI (Copley Scientific, Nottingham , UK), apparatus C, as described in European Pharmacopoeia 8.0. (2014). A 20 mg sample of each DPI formulation (comparative example and in accordance with example 2), previously sieved through a 355 mm diameter stainless steel sieve, was weighed into a size 3 HPMC capsule (Quali-V-I, Qualicaples, Madrid, Spain) and placed in MsLI using a dry powder inhaler RS.01 (RPC Plastiape, Osnago, Italy) mounted on the inhalation port using its adapter (n = 3).

A sedimentation flow rate of 100 ± 5 L/min measured with a DFM3 flow meter (Copley Scientific, Nottingham, UK) was obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series with a TPK critical flow controller (Copley Scientific , Nottingham, UK).

At this flow rate, the critical diameters between each MsLI stage were 10.0, 5.3, 2.4, 1.3 and 0.4 mm. The micro-diaphragm collection (MOC) filter (i.e., stage 5) contained a 9 cm Fluoropore PTFE membrane with 0.45 mm pore size adhered to a high-density polyethylene backing (Merck Millipore, Darmstadt, Germany). A critical flow controller was used to achieve a settling time of 2.4 s at 100 L/min and a critical flow rate of P3/P2 < 0.5 as required by European Pharmacopoeia 8.0. (2014).

After impaction, the top four stages of MsLI were subjected to a first wash using 20 mL of 0.5% w/v prefill. poloxamer 407 in ultrapure water/isopropanol (60:40 v/v) as dilution phase, a second wash with 25 ml DMF and a third wash with dilution phase, adjusted to 100.0 ml and sonicated in for 30 minutes. Drug deposition into the capsule, device, induction port, and MOC filter was determined after dissolution in 100.0 mL of dilution phase and sonication for 30 min. The impacted mass at each stage was determined by quantifying the cisplatin content using a validated electrothermal atomic absorption spectrometry (ETAAS) method described by Levet el al (Levet, Int J Pharm 2016).

The results were then plotted using Copley Inhaler Testing Data Analysis Software1 (Copley Scientific, Nottingham, UK) to obtain an FPD of less than 5 µm. This was done by interpolating the extracted mass depending on the critical diameter of the corresponding stage. FPF was expressed as a percentage of the nominal dose.

In fig. 1 shows the FPF values (%) of a comparative example (composition F5 in Levet el al and examples 1 and 2 (mean ± SD, n = 2-3), demonstrating the superiority of the cisplatin compositions described in the present invention compared to other DPIs -triglyceride-based formulations with cisplatin regarding pulmonary deposition.

Example 4 Analysis of the dissolution rate of cisplatin from dry powder formulations of cisplatin prepared in accordance with examples 1 and 2

The dissolution properties of DPI formulations were determined by applying the method described by Levet et al (Levet et al, Int J Pharm 2016). This method is based on the stirrer-over-disc method of USP39 and uses a modified Type V dissolution apparatus for transdermal patches. The release profile of cisplatin was determined from the total inhaled fraction (dae less than or equal to 5 μm) of the DPI formulation selected using a Fast Screening Impactor (FSI, Copley Scientific, Nottingham, UK). The corresponding mass of each DPI formulation, equivalent to a precipitated dose of 3 mg of cisplatin, was weighed into a size 3 HPMC capsule (Quali-V-I Qualicaples, Madrid, Spain). It was then applied using a DPI device RS.01 (RPC Plastiape) onto a hydrophobic Fluoropore® PTFE membrane filter with 0.45 mm pore size (Merck Millipore, Darmstadt, Germany) with FSI (2.4 s, 100 l/min ), equipped with a corresponding preseparator insert. The Fluoropore filter, with the deposited powder facing upward, was then covered with a 0.4 mm hydrophilic polycarbonate Isopore® filter (Merck-Milipore, Germany) and secured to a convex PTFE disc assembly (Copley, Nottingham, UK) using PTFE clamps and mesh. The disc assembly was then immersed in the dissolution vessel of the AT7 dissolution apparatus (Sotax AG, Aesch, Switzerland) using 400 ml of modified simulated pulmonary fluid (mSLF) (Son and McConville, 2009), which is a medium simulating the composition of electrolytes and surfactants. lung substances.

The dissolution test was carried out according to sufficient dissolution conditions at 37 ± 0.2 °C, pH 7.35 ± 0.05. The blades, mounted 25 ± 2 mm between the blade and the center of the disc assembly, were set at a rotation speed of 50 ± 4 rpm. 2.0 mL samples were filtered through 0.22 mm pore size cellulose acetate syringe filters (VWR, Leuven, Belgium) at predetermined time points ranging from 2 min to 24 h and replaced with 2.0 mL of free, prewarmed mSLF.

At the end of the dissolution assay, the disc assembly was opened into the dissolution vessel and sonicated for 30 min to achieve 100% cisplatin dissolution value.

In fig. 2 shows the release profiles of cisplatin from the inhalable fraction of DPI formulations prepared in accordance with Examples 1 and 2, relative to a comparative formulation consisting of cisplatin microparticles only.

Example 5. Preparation of dry powder composition of insulin No. 1

Insulin was first suspended in isopropanol (2% w/v) and the powder dispersion was obtained by ultrasonication for 10 min in a Branson 2510 40 kHz bath. The particle size was then reduced using an EmulsiFlex-C5 high-pressure homogenizer (Aves-tin Inc., Ottawa, Canada). First, low pressure homogenization cycles were performed to pre-grind on the insulin slurry to further reduce particle size (10 cycles at 7,000 psi and 10 cycles at 12,000 psi). Finally, HPH was then applied for 30 cycles at 24,000 psi. inch. These cycles were performed by recirculating the treated suspension directly into the sample reservoir (closed loop). Because HPH causes an increase in sample temperature (30°C increase after 20 cycles at 24,000 psi), all operations were performed using a heat exchanger mounted upstream of the homogenizing valve, with the sample temperature maintained at 5 ± 1°C.

At the end of the process, an aliquot was removed from the suspension to measure the particle size distribution (PSD) of the insulin microcrystals using laser diffraction (see below).

Hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA) dissolved in heated isopropanol were then added to the microcrystalline suspension to obtain a final insulin concentration of 1.0% w/v. and a mixture (99: 1 w/w) of hydrogenated castor oil/TPGS 1.0% w/v. and spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

The geometric PSD of bulk insulin, insulin microparticles produced by the size reduction method, and insulin dry powder formulation was measured as suspended and individualized particles, and the resulting value was within the range of the present invention. For this purpose, a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used as described in Example 1.

Example 6. Preparation of dry powder composition of insulin No. 2

Insulin was first suspended in isopropanol (2% w/v) and the powder dispersion was obtained by ultrasonication for 10 min in a Branson 2510 40 kHz bath. The particle size was then reduced using an EmulsiFlex-C5 high-pressure homogenizer (Aves-tin Inc., Ottawa, Canada). First, low pressure homogenization cycles were performed to pre-grind on the insulin slurry to further reduce particle size (10 cycles at 7,000 psi and 10 cycles at 12,000 psi). Finally, HPH was then applied for 30 cycles at 24,000 psi. inch. These cycles were performed by recirculating the treated suspension directly into the sample reservoir (closed loop). Because HPH causes an increase in sample temperature (30°C increase after 20 cycles at 24,000 psi), all operations were performed using a heat exchanger mounted upstream of the homogenizing valve, with the sample temperature maintained at 5 ± 1°C.

At the end of the process, an aliquot was removed from the suspension to measure the particle size distribution (PSD) of the insulin microcrystals using laser diffraction (see below).

Hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA) dissolved in heated isopropanol were then added to the microcrystalline suspension to obtain a final insulin concentration of 1.5% w/v. and a mixture (99: 1 w/w) of hydrogenated castor oil/TPGS 0.5% w/v. and spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

The geometric PSD of bulk insulin, insulin microparticles produced by the size reduction method, and insulin dry powder formulation was measured as suspended and individualized particles, and the resulting value was within the range of the present invention. For this purpose, a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used as described in Example 1.

Example 7. Preparation of dry powder composition of budesonide for inhalation No. 1

Budesonide (1% w/v) was first dissolved in isopropanol under magnetic stirring. Hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA) dissolved in heated isopropanol were then added to the microcrystalline suspension to obtain a final budesonide concentration of 1.0% w/v. and a mixture (99: 1 w/w) of hydrogenated castor oil/TPGS 1.0% w/v. and spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

The geometric PSD of the dry powder formulation of bulk budesonide was measured as suspended and individualized particles, and the resulting value was within the range of the present invention. For this purpose, a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used as described in Example 1.

Example 8. Preparation of dry powder composition of budesonide for inhalation No. 2

Budesonide (1.5% w/v) was first dissolved in isopropanol under magnetic stirring. Hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA) dissolved in heated isopropanol were then added to the microcrystalline suspension to obtain a final budesonide concentration of 1.5% w/v. and a mixture (99: 1 w/w) of hydrogenated castor oil/TPGS 0.5% w/v. and spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

The geometric PSD of the dry powder formulation of bulk budesonide was measured as suspended and individualized particles, and the resulting value was within the range of the present invention. For this purpose, a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used as described in Example 1.

Examples 9. Preparation of dry powder composition of budesonide No. 3

10 g micronized budesonide, 9.9 g hydrogenated castor oil and 0.1 g TPGS were mixed until homogeneous in a Turbula® mixer (Willy A. Bachofen AG, Muttenz, Switzerland). The homogeneous mixture was then extruded through a twin-screw extruder (Process-11, Thermo Fischer Scientific, Massachusetts, USA) at an appropriate temperature to obtain a homogeneous lipid matrix. The extrudate was then cut into large beads and the beads were placed in an incubator under appropriate storage conditions to convert the lipid matrix into a stable polymorphic material. Finally, the granules were milled using a jet mill (with appropriate granule feed rate, injection pressure, and attrition pressure) to produce microparticles for inhalation (DPI) for human use.

Example 10. Preparation of dry powder composition of pemetrexed for inhalation No. 1

Briefly, crude pemetrexed disodium microcrystals (heptahydrate form) from bulk powder (Carbosynth Limited, Berkshire, UK) were first suspended in 50 ml isopropanol to achieve a concentration of 1% w/v. in the presence of 0.05% w/v. TPGS (Sigma-Aldrich, St-Louis, USA) and reduced in size by high-speed (10 min at 24,000 rpm) (X620 motor and T10 dispersing shaft, Ingenieurbüro CAT M. Zipperer GmbH, Staufen, Germany) homogenization under high pressure (EmulsiFlex-C3 high pressure homogenizer, Avestin Inc., Ottawa, Canada) at 25,000 psi. inch for 20 grinding cycles. The heat exchanger was connected to a homogenizing valve and maintained at +5°C using an F32-MA cooling circulator (Julabo GmbH, Seelbach, Germany). At the end of the process, an aliquot was removed from the suspension to measure the particle size distribution (PSD) of pemetrexed microcrystals using laser diffraction (see below).

Then 1% w/v. hydrogenated castor oil (BASF, Ludwigshafen‎, Germany) was dissolved in a heated (50°C) microcrystal suspension and spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

The geometric PSD of bulk pemetrexed, microparticles of pemetrexed produced by the size reduction method, and dry powder formulation of pemetrexed were measured as suspended and individualized particles, and the resulting value was within the range of the present invention. For this purpose, a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used as described in Example 1.

Example 11 Settlement Rate Analysis of Pemetrexed Dry Powder Formulation Prepared in Example 10

The formulation according to Examples 10 was analyzed for in vitro lung deposition type and FPF value.

The FPF value, the percentage related to the extracted dose of pemetrexed-based particles with an aerodynamic diameter (dae) of less than 5 μm and an aerodynamic PSD characterized by MMAD, was determined using NGI (Copley Scientific, Nottingham, UK), apparatus E, as described in the European Pharmacopoeia 8.0. (2014). A 20 mg sample of the DPI formulation (according to Example 10 and a comparative formulation consisting only of pemetrexed microparticles), previously sifted through a 355 mm diameter stainless steel sieve, was weighed into a size 3 HPMC capsule (Quali-V-I, Qualicaples, Madrid, Spain) and placed into the NGI using a dry powder inhaler RS.01 (RPC Plastiape, Osnago, Italy) mounted on the inhalation port using its adapter (n = 3).

A sedimentation flow rate of 100 ± 5 L/min measured with a DFM3 flow meter (Copley Scientific, Nottingham, UK) was obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series with a TPK critical flow controller (Copley Scientific , Nottingham, UK).

At this flow rate, the critical diameters between each NGI stage were 6.12, 3.42, 2.18, 1.31, 0.72, 0.40, and 0.24 mm. A critical flow controller was used to achieve a settling time of 2.4 s at 100 L/min and a critical flow with a P3/P2 ratio < 0.5 as required by European Pharmacopoeia 8.0 (2014).

After impaction, the pemetrexed mass deposited in the capsule, device, induction port, preseparator, 7 stages and NGI MOC was collected using ultrapure water/DMF (30:70 v/v) as the dilution phase and were subjected to ultrasonic treatment for 30 min. The impacted mass at each stage was determined by quantifying the pemetrexed content using a validated HPLC. The chromatographic system (HP 1200 series, Agilent Technologies, Diegem, Belgium) was equipped with a quaternary pump, an autosampler, and a diode array detector. Separation was performed on a Hypersil Gold C18 reverse phase column (5 mm, 250 mm x 4.6 mm) (Thermo Fisher Scientific, Waltham, USA). The mobile phase consisted of ultrapure water/acetonitrile (86:14) acidified with 0.4% formic acid, which was 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 at 30°C, and the assay time was 15 min.

The results were then plotted using Copley Inhaler Testing Data Analysis Software1 (Copley Scientific, Nottingham, UK) to obtain an FPD of less than 5 µm. This was done by interpolating the extracted mass depending on the critical diameter of the corresponding stage. FPF was expressed as a percentage of the nominal dose.

In fig. 3 presents the FPF value of Example 10 relative to a comparative microparticulate pemetrexed formulation only (mean ± SD, n = 3 and 1, respectively), demonstrating the superiority of Example 10 relative to the standard DPI formulation in terms of pulmonary deposition.

Example 12 Analysis of the dissolution rate of pemetrexed from the dry powder formulation of pemetrexed prepared in accordance with Examples 10

The dissolution properties of the DPI formulation were determined by using an adapted method based on the method described by Pilcer et al (Pilcer et al, J Pharm Sci 2013). A dissolution system (Copley Scientific, Nottingham, UK) specifically designed for DPI release profile studies was used in conjunction with the stirrer-over-disc method (Eur.Ph. 7). To study the release profiles of particles that are deposited in the lungs, fractionation of the pemetrexed-containing formulation was first performed using NGI. The cup in step 3 was selected to be equipped with a removable disk insert for collecting particles. Of particular interest is that at the selected inhalation rate (100 L/min for 2.4 s), the critical diameters at stage 3 ranged from 2.18 to 3.42 μm, allowing the selection of lung-targeted particles. Capsules containing the appropriate amount of formulation according to Example 10 were weighed to collect approximately 6 mg of pemetrexed in step 3. The disk insert was then coated with a polycarbonate membrane (pore size 0.4 μm) (Merck Millipore) and placed in a paddle dissolution apparatus (Erweka DT6; ERWEKA GmbH, Heusenstamm, Hesse, Germany) filled with 400 ml mSLF (Son and McConville, 2009), a medium simulating the electrolyte and surfactant composition of the lung.

The dissolution test was carried out according to sufficient dissolution conditions at 37 ± 0.2 °C, pH 7.35 ± 0.05. The blades, mounted 25 ± 2 mm between the blade and the center of the disc assembly, were set at a rotation speed of 50 ± 4 rpm. 2.0 mL samples were filtered through 0.22 mm pore size cellulose acetate syringe filters (VWR, Leuven, Belgium) at predetermined time points ranging from 2 min to 24 h and replaced with 2.0 mL of free, prewarmed mSLF.

At the end of the dissolution assay, the disc assembly was opened into the dissolution vessel and sonicated for 30 min to achieve 100% pemetrexed dissolution value.

In fig. 4 shows the release profiles of pemetrexed from the inhalable fraction of the DPI formulation prepared in accordance with Example 10 relative to a comparative formulation consisting of pemetrexed microparticles only (mean ± SD, n = 3).

The similarity factor f2 was used to compare the two dissolution profiles (Shah et al, Pharm Res 1998). The curves were significantly different (f2 < 50). Moreover, the total release values at all time points from Example 10 were significantly lower compared to the values from pemetrexed microcrystals (p < 0.05, t test), for example, after 1 hour, 53 ± 9% and 97, respectively. 9 ± 0.9% (P < 0.01), indicating a controlled release profile of pemetrexed from the composition of the present invention.

Example 13. Preparation of dry powder composition of insulin No. 3

Insulin (Sigma-Aldrich) was first suspended in isopropanol (1% w/v) using a magnetic stirrer, and the powder dispersion was prepared by sonication for 10 min in a Branson 2510 40 kHz bath. Particle size was then reduced using an EmulsiFlex-C3 high-pressure homogenizer (Avestin Inc., Ottawa, Canada) for 30 cycles at 22,000 psi. inch. These cycles were performed by recirculating the treated suspension directly into the sample reservoir (closed loop). Since HPH causes an increase in sample temperature, all operations were performed using a heat exchanger installed in front of the homogenizing valve, and the sample temperature was maintained at 5 ± 1 °C.

At the end of the process, an aliquot was removed from the suspension to measure the particle size distribution (PSD) of the insulin microcrystals using laser diffraction (see below).

Then 1% w/v. hydrogenated castor oil (BASF, Ludwigshafen‎, Germany) was dissolved in a heated (50°C) microcrystal suspension and spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

The geometric PSD of bulk insulin, insulin microparticles produced by the size reduction method, and insulin dry powder formulation was measured as suspended and individualized particles, and the resulting value was within the range of the present invention. For this purpose, a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used as described in Example 1.

Example 14 Settlement Rate Analysis of Dry Powder Insulin Formulations Prepared in Example 13

The formulation according to Example 13 was analyzed for FPF value.

The FPF value, the percentage value related to the extracted dose of insulin-based particles with dae less than 5 μm, was determined using a Fast Screening Impactor (FSI) (Copley Scientific, Nottingham, UK). FSI uses a two-stage separation method in which the first large non-inhaled boluses are captured in a liquid separator, followed by a fine-cut impaction stage at 5 µm (i.e. according to FPF). A 10 mg sample of DPI formulations (according to example 13), previously sieved through a 355 mm diameter stainless steel sieve, was weighed into a size 3 HPMC capsule (Quali-V-I, Qualicaples, Madrid, Spain) and placed in FSI using dry powder inhaler RS.01 (RPC Plastiape, Osnago, Italy) mounted on the inhalation port using its adapter (n = 3).

A sedimentation flow rate of 100 ± 5 L/min measured with a DFM3 flow meter (Copley Scientific, Nottingham, UK) was obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series with a TPK critical flow controller (Copley Scientific , Nottingham, UK). A critical flow controller was used to achieve a deposition time of 2.4 s at a flow rate of 100 L/min.

After impaction, the insulin mass is deposited in the capsule, in the device, in the induction port, in the preseparator and on a Fluoropore PTFE membrane measuring 9 cm and pore size 0.45 mm, glued to a high-density polyethylene backing (Merck Millipore, Darmstadt, Germany) , were collected using 0.01 M HCl as a dilution phase and sonicated for 30 min. The impacted mass at each stage was determined by quantifying the insulin content using the HPLC method described in European Pharmacopoeia 9.2. (2017). FPF was expressed as a percentage of the nominal dose.

In fig. 5 shows the FPF values from Example 13 (mean ± SD) demonstrating the high lung deposition rate of the insulin-based DPI composition described in the present invention.

Example 15 Analysis of the dissolution rate of insulin from dry insulin powder formulations prepared in accordance with Example 13

The dissolution properties of the DPI composition according to example 13 were determined by using an adapted method based on the method described by Depreter et al (Depreter et al, Eur J Pharm Biopharm 2012). A dissolution system (Copley Scientific, Nottingham, UK) specifically designed for DPI release profile studies was used in conjunction with the stirrer-over-disc method (Eur.Ph. 7). To study the release profiles of particles that are deposited in the lungs, fractionation of the insulin-containing formulation was first performed using NGI. The cup in step 3 was selected to be equipped with a removable disk insert for collecting particles. Of particular interest is that at the selected inhalation rate (100 L/min for 2.4 s), the critical diameters at stage 3 ranged from 2.18 to 3.42 μm, allowing the selection of lung-targeted particles. Capsules with the appropriate amount of formulation according to Example 13 were weighed to collect about 3 mg of insulin in step 3. The disk insert was then coated with a polycarbonate membrane (pore size 0.4 μm) (Merck Millipore) and placed in a paddle dissolution apparatus (Erweka DT6; ERWEKA GmbH, Heusenstamm, Hesse, Germany) filled with 400 ml of 0.01 mM phosphate-buffered saline (PBS) at pH 7.4.

The dissolution test was carried out according to sufficient dissolution conditions at 37 ± 0.2 °C, pH 7.35 ± 0.05. The blades, mounted 25 ± 2 mm between the blade and the center of the disc assembly, were set at a rotation speed of 50 ± 4 rpm. 5.0 mL samples were withdrawn at specified time points ranging from 2 min to 24 h and replaced with 5.0 mL of neat prewarmed PBS.

At the end of the dissolution assay, the disk assembly was opened into the dissolution vessel and sonicated for 30 min to achieve 100% insulin dissolution value.

Samples were lyophilized (Christ Epsilon 1–6) in the presence of 3% w/v. trehalose. The lyophilisates were dissolved in 500 μl of 0.02 N HCl and injected into the HPLC system using the method described in European Pharmacopoeia 9.2. (2017).

In fig. 6 shows the insulin release profiles from the inhaled fraction of DPI formulations prepared in accordance with Example 13 (up to 240 min, mean ± SD, n = 2) (A), relative to the comparative formulations described by Depreter et al (up to 180 min) (B).

The cumulative release values at various time points from Example 13 were lower compared to those of the two formulations from Depreter et al, for example, after 1 hour, respectively, about 60% compared to about 100%, which demonstrates the advantages of the insulin compositions described in the present invention, from the point of view of control of insulin release both in the case of insulin microcrystals and in the case of lipid-coated insulin microcrystals described by Depreter et al (F1 and F2 in Fig. 6, respectively).

Example 16 Preparation of Cisplatin No. 3 Dry Powder Formulation and Comparative Example

First, cisplatin (Umicore, Hanau-Wolfgang, Germany) was suspended in 50 ml ethanol to achieve a concentration of 5% w/v. and reduced in size by high-speed (10 min at 24,000 rpm) (X620 motor and T10 dispersing shaft, Ingenieurbüro CAT M. Zipperer GmbH, Staufen, Germany) high-pressure homogenization using an EmulsiFlex-C3 high-pressure homogenizer (Avestin Inc., Ottawa, Canada) for 40 cycles at 20,000 psi. inch. These cycles were performed by recirculating the treated suspension directly into the sample reservoir (closed loop). Since HPH causes an increase in sample temperature, all operations were performed using a heat exchanger installed in front of the homogenizing valve, and the sample temperature was maintained at 15 ± 1 °C.

At the end of the process, an aliquot was removed from the suspension to measure the PSD of cisplatin microcrystals using laser diffraction (see below).

Hydrogenated castor oil (BASF, Ludwigshafen, Germany) and TPGS (Sigma-Aldrich, St-Louis, USA) (or tristearin in the case of the comparative example) dissolved in heated isopropanol were then added to the microcrystalline suspension to obtain a final concentration of cisplatin 2, 0% w/v and a mixture (99: 1 w/w) of triglyceride/TPGS 2.0% w/v. and spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI formulation for human use. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

The geometric PSD of bulk cisplatin, microparticles of cisplatin obtained by the size reduction method, and dry powder formulation of cisplatin were measured as suspended and individualized particles, and the obtained value was within the range of the present invention. For this purpose, a Mastersizer 3000 laser diffractometer (Malvern Instruments Ltd., Worcestershire, UK) coupled to a Hydro MV dispenser equipped with a 40 W ultrasonic probe (Malvern Instruments Ltd.) was used as described in Example 1.

Example 17. Analysis of the sedimentation rate of a dry powder composition of cisplatin obtained in accordance with example 6 16

The formulations of Examples 16 were analyzed for in vitro lung deposition type and FPF value.

FPF, the percentage related to the extracted dose of cisplatin-based particles with a dae value of less than 5 μm and an aerodynamic PSD characterized by MMAD, was determined using NGI (Copley Scientific, Nottingham, UK), apparatus E, as described in European Pharmacopoeia 8.0. (2014). A 20 mg sample of DPI formulations (according to Example 17), previously sieved through a 355 mm diameter stainless steel sieve, was weighed into a size 3 HPMC capsule (Quali-V-I, Qualicaples, Madrid, Spain) and placed in FSI using dry powder inhaler RS.01 (RPC Plastiape, Osnago, Italy) mounted on the inhalation port using its adapter (n = 3).

A sedimentation flow rate of 100 ± 5 L/min measured with a DFM3 flow meter (Copley Scientific, Nottingham, UK) was obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series with a TPK critical flow controller (Copley Scientific , Nottingham, UK).

At this flow rate, the critical diameters between each NGI stage were 6.12, 3.42, 2.18, 1.31, 0.72, 0.40, and 0.24 mm. A critical flow controller was used to achieve a settling time of 2.4 s at 100 L/min and a critical flow rate of P3/P2 < 0.5 as required by European Pharmacopoeia 8.0. (2014).

After impaction, the bulk of cisplatin deposited in the capsule, device, induction port, preseparator, 7 steps, and MOC NGI was collected using DMF as the dilution phase and sonicated for 30 min. Impacted mass at each stage was determined by quantifying cisplatin content using the validated ETAAS method described by Levet et al (Levet et al, Int J Pharm 2016).

The results were then plotted using Copley Inhaler Testing Data Analysis Software1 (Copley Scientific, Nottingham, UK) to obtain an FPD of less than 5 µm. This was done by interpolating the extracted mass depending on the critical diameter of the corresponding stage. FPF was expressed as a percentage of the nominal dose.

The results obtained were combined with the results obtained in Example 3, demonstrating the superiority of the cisplatin compositions described in the present invention compared to other triglyceride-based cisplatin DPI formulations in terms of pulmonary deposition.

Example 18 Analysis of the dissolution rate of cisplatin from a dry powder formulation of cisplatin prepared in accordance with Examples 16

The dissolution properties of the DPI formulation were determined by using an adapted method based on the method described by Pilcer et al (Pilcer et al, J Pharm Sci 2013). A dissolution system (Copley Scientific, Nottingham, UK) specifically designed for DPI release profile studies was used in conjunction with the stirrer-over-disc method (Eur.Ph. 7). To study the release profiles of particles that are deposited in the lungs, fractionation of the cisplatin-containing formulation was first performed using NGI. The cup in step 3 was selected to be equipped with a removable disk insert for collecting particles. Of particular interest is that at the selected inhalation rate (100 L/min for 2.4 s), the critical diameters at stage 3 were in the range from 2.18 to 3.42 μm, which made it possible to select particles with a targeted effect on the lung, from a capsule filled with an appropriate amount of the formulations according to Example 16 to precipitate about 2 mg of cisplatin in step 3. The disk insert was then coated with a polycarbonate membrane (pore size 0.4 μm) (Merck Millipore) and placed in a paddle dissolution apparatus (Erweka DT6; ERWEKA GmbH, Heusenstamm, Hesse, Germany) filled with 400 ml mSLF (Son and McConville, 2009), a medium simulating the electrolyte and surfactant composition of the lung.

The dissolution test was carried out according to sufficient dissolution conditions at 37 ± 0.2 °C, pH 7.35 ± 0.05. The blades, mounted 25 ± 2 mm between the blade and the center of the disc assembly, were set at a rotation speed of 50 ± 4 rpm. 2.0 mL samples were filtered through 0.22 mm pore size cellulose acetate syringe filters (VWR, Leuven, Belgium) at predetermined time points ranging from 2 min to 24 h and replaced with 2.0 mL of free, prewarmed mSLF.

At the end of the dissolution assay, the disc assembly was opened into the dissolution vessel and sonicated for 30 min to achieve 100% cisplatin dissolution value.

In fig. 7 shows the release profiles of cisplatin from the inhaled fraction of the DPI formulation prepared in accordance with Example 16 relative to a comparative formulation consisting of microparticulate cisplatin (mean ± SD, n = 3).

The similarity factor f2 was used to compare the two dissolution profiles (Shah et al, Pharm Res 1998). The curves were significantly different (f2 < 50). Moreover, the total release values at all time points from Example 16 were significantly lower compared to the values from cisplatin microcrystals (p < 0.05, t test), for example, after 4 hours, 57 ± 4% and 76 ± 5% (p < 0.01), indicating a controlled release profile of cisplatin from the composition of the present invention.

Example 19. Preparation of dry powder composition of budesonide for inhalation No. 4

0.1500% w/v budesonide, 2.8215% w/v. hydrogenated castor oil (BASF, Ludwigshafen‎, Germany) (or tristearin for comparative examples BUD-TS4) and 0.0285% w/v. TPGS (Sigma-Aldrich, St-Louis, USA) was dissolved in hot isopropanol (65°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 20. Preparation of dry powder composition of budesonide for inhalation No. 5

0.600% w/v budesonide, 2.376% w/v. hydrogenated castor oil (BASF, Ludwigshafen, Germany) (or tristearin for comparative examples BUD-TS5) and 0.024% w/v. TPGS (Sigma-Aldrich, St-Louis, USA) was dissolved in hot isopropanol (65°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 21. Preparation of dry powder composition of budesonide for inhalation No. 6

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) was dissolved in hot isopropanol (65°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 22 Settlement Rate Analysis of Dry Powder Formulations of Budesonide Prepared According to Examples 19 and 20

The formulations of Examples 19 and 20 were analyzed for in vitro lung deposition type and fine particle fraction value. Comparative examples of compositions 19 and 20, i.e. BUD-TS4 and BUD-TS5, respectively, were prepared using tristearin (TS) (Tokyo Chemical Company, Tokyo, Japan) instead of hydrogenated castor oil according to the same corresponding protocols.

FPF, the percentage related to the extracted dose of budesonide-based particles with a dae value of less than 5 μm and an aerodynamic PSD characterized by MMAD, was determined using NGI (Copley Scientific, Nottingham, UK), apparatus E, as described in European Pharmacopoeia 8.0. (2014). A 10 mg sample of DPI formulations (according to examples 19 and 20 and comparative powders BUD-TS4 and BUD-TS5), previously sifted through a stainless steel sieve with a diameter of 355 mm, was weighed into a HPMC capsule size 3 (Quali-V-I, Qualicaples, Madrid , Spain) and placed into the NGI using a dry powder inhaler RS.01 (RPC Plastiape, Osnago, Italy) mounted on the inhalation port using its adapter (n = 3).

A sedimentation flow rate of 100 ± 5 L/min measured with a DFM3 flow meter (Copley Scientific, Nottingham, UK) was obtained using two HCP5 air pumps (Copley Scientific, Nottingham, UK) connected in series with a TPK critical flow controller (Copley Scientific , Nottingham, UK).

At this flow rate, the critical diameters between each NGI stage were 6.12, 3.42, 2.18, 1.31, 0.72, 0.40, and 0.24 mm. A critical flow controller was used to achieve a settling time of 2.4 s at 100 L/min and a critical flow rate of P3/P2 < 0.5 as required by European Pharmacopoeia 8.0. (2014).

After impaction, the mass of budesonide deposited in the capsule, device, induction port, preseparator, 7 stages and MOC NGI was collected using 0.5% w/v. poloxamer 407 in a mixture of ultrapure water: isopropanol in a ratio of 60: 40 (v/v) as a dilution phase and subjected to ultrasonic treatment for 30 min at 60°C. The solutions were then filtered through Minisart regenerated cellulose syringe filters with a pore size of 0.45 μm (Sartorius Stedim Biotech GmbH, Germany). The impacted mass at each stage was determined by quantifying the budesonide content using a validated HPLC method. The chromatographic system (HP 1200 series, Agilent Technologies, Diegem, Belgium) was equipped with a quaternary pump, an autosampler, and a diode array detector. Separation was carried out on an Alltima C18 reverse phase column (5 mm, 150 mm x 4.6 mm) (Hichrom, Theale, UK). The mobile phase consisted of a mixture of phosphate buffer: acetonitrile (65: 35 v/v), pH = 3.20, which was delivered at a flow rate of 1.5 ml/min. Quantification was performed at 245 nm. The injection volume was 100 μL, the temperature was set at 40°C, and the assay time was 22 min.

The results were then plotted using Copley Inhaler Testing Data Analysis Software1 (Copley Scientific, Nottingham, UK) to obtain an FPD of less than 5 µm. This was done by interpolating the extracted mass depending on the critical diameter of the corresponding stage. FPF was expressed as a percentage of the nominal dose.

In fig. 8 shows the FPF values from Examples 19 and 20 relative to Comparative Examples BUD-TS4 and BUD-TS5 (mean ± SD, n = 3). (***) p < 0.001, t-test demonstrating the superiority of the budesonide compositions described in the present invention compared to other triglyceride-based budesonide DPI formulations in terms of pulmonary deposition.

Example 23 Analysis of the dissolution rate of budesonide from dry powder formulations of budesonide prepared in accordance with Examples 19, 20 and 21

The dissolution properties of the DPI formulations according to Examples 19, 20 and 21 and the comparative budesonide powder (i.e. micronized budesonide) were determined by using a method adapted from the method described by Pilcer et al (Pilcer et al, J Pharm Sci 2013 ). A comparative micronized budesonide powder was prepared by spray drying (Mini-Spray Dryer B-290, Büchi Labortechnik AG, Flawil, Switzerland) 3% w/v. a solution of budesonide in isopropanol to obtain a DPI composition for use in humans.

A dissolution system (Copley Scientific, Nottingham, UK) specifically designed for DPI release profile studies was used in conjunction with the stirrer-over-disc method (Eur.Ph. 7). To study the release profiles of particles that are deposited in the lungs, fractionation of the budesonide-containing formulation was first performed using NGI. The cup in step 2 was selected to be equipped with a removable disk insert for collecting particles. Of particular interest is that at the selected inhalation rate (100 L/min for 2.4 s), the critical diameters at stage 2 ranged from 6.12 to 3.42 μm, which allowed the selection of particles with a targeted effect on the lung, from a capsule filled with the appropriate amount of formulation (according to micronized budesonide and Examples 20, 21 and 22) to precipitate about 500 μg of budesonide in step 3. The disc insert was then covered with a polycarbonate membrane (pore size 0.4 μm) (Merck Millipore) and placed in a paddle dissolution apparatus (Erweka DT6; ERWEKA GmbH, Heusenstamm, Hesse, Germany) filled with 400 ml of 0.01 mM phosphate-buffered saline (PBS) at pH 7.4.

The dissolution test was carried out according to sufficient dissolution conditions at 37 ± 0.2 °C, pH 7.35 ± 0.05. The blades, mounted 25 ± 2 mm between the blade and the center of the disc assembly, were set at a rotation speed of 50 ± 4 rpm. 2.0-mL samples were filtered through 0.22-mm cellulose acetate syringe filters (VWR, Leuven, Belgium) at predetermined time points ranging from 2 min to 24 h and replaced with 2.0 mL of neat prewarmed PBS.

The value of 100% dissolution of budesonide corresponds to the mass deposited at stage 3.

In fig. 9 shows the release profiles of budesonide from the inhaled fraction of a comparative example of micronized budesonide and DPI formulations prepared in accordance with Examples 19, 20 and 21 (n = 1), which indicate a controlled release profile of budesonide from the compositions of the present invention and the ability to modulate the release profile by modulation of the drug/lipid ratio.

Example 24. Preparation of dry powder composition of paclitaxel for inhalation No. 1

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) was dissolved in hot ethanol (50°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 25. Preparation of dry powder composition of paclitaxel for inhalation No. 2

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) was dissolved in hot ethanol (50°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 26. Preparation of dry powder composition of paclitaxel for inhalation No. 3

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) was dissolved in hot ethanol (50°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 27. Preparation of dry powder composition of paclitaxel for inhalation No. 4

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) was dissolved in hot ethanol (50°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 28. Preparation of dry powder composition of paclitaxel for inhalation No. 5

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) was dissolved in hot ethanol (50°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 29. Preparation of dry powder composition of paclitaxel for inhalation No. 6

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) was dissolved in hot ethanol (50°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain a DPI composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 30. Preparation of dry powder composition of paclitaxel for inhalation No. 7

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) was dissolved in hot ethanol (50°C) with magnetic stirring and the hot solution was spray dried using a Mini-Spray Dryer B-290 (Büchi Labortechnik AG, Flawil, Switzerland) to obtain DPI- composition for use in humans. The operating parameters used in spray drying were as follows: feed rate 3.0 g/min, inlet temperature 70°C, nozzle 0.7 mm, nozzle cap 1.5 mm, compressed air at 800 L/min and flow drying air 35 m³/h. The apparatus was equipped with a B-296 dehumidifier (Büchi Labortechnik AG) to maintain a relative humidity of 50% RH during spray drying.

Example 31. Preparation of dry powder composition of budesonide for inhalation No. 7

First, dry powder budesonide inhalation formulation No. 4 was prepared according to Example 19. Then, 10 g dry powder budesonide inhalation formulation No. 4 and 30 g lactose (Respitose® SV003, DFE Pharma, Goch, Germany) were mixed until homogeneous in a Turbula mixer ® (Willy A. Bachofen AG, Muttenz, Switzerland) to obtain a dry powder formulation for inhalation based on a carrier.

It should be understood that the present invention is not limited to the embodiments described and that changes may be made without departing from the scope of the appended claims.

Claims (28)

1. Dry powder inhalation composition for administration to the lungs, containing: - at least one active pharmaceutical ingredient (API) absorbed by the lungs for systemic or local therapy, and - a lipid matrix containing at least one triglyceride selected from the group consisting of monohydroxystearin, dihydroxystearin, trihydroxystearin and mixtures thereof, and wherein said composition has a mass ratio between at least one API and said lipid matrix: API/lipid matrix in the range from 0.1/99.9 to 88/12, contains from 0.1 wt.% to 95 wt.% of the specified at least one API relative to the weight of the specified dry powder inhalation composition, wherein said composition is in the form of fine particles having a mass median aerodynamic diameter (MMAD) that is less than or equal to 6 microns. 2. The dry powder inhalation composition of claim 1, having a weight ratio between said at least one API and said lipid matrix: API/lipid matrix in the range of 10/90 to 88/12, preferably from 15/85 to 85/15 , preferably from 25/75 to 75/25, more preferably from 30/70 to 70/30, in particular from 40/60 to 60/40, for example about 50/50, based on the total weight of the dry powder inhalation composition. 3. The dry powder inhalation composition according to claim 1 or 2, wherein said at least one triglyceride is hydrogenated castor oil. 4. Dry powder inhalation composition according to any one of paragraphs. 1-3, wherein said API is a small chemical molecule having a solubility in alcohols of at least 0.1% w/v, more preferably at least 0.5% w/v, more preferably at least at least 1% w/v, more particularly at least 5% w/v. 5. Dry powder inhalation composition according to claim 4, containing from 1% wt. up to 85% wt. of said at least one API, relative to the weight of the dry powder inhalation composition. 6. Dry powder inhalation composition according to any one of paragraphs. 1-3, wherein said API is a small chemical molecule having a solubility in alcohols of less than 0.5% w/v, more preferably less than 0.1% w/v, more preferably less than 0.01% w/v ./about. 7. Dry powder inhalation composition according to claim 6, containing from 1% wt. up to 85% wt. of said at least one API, relative to the weight of the dry powder inhalation composition. 8. Dry powder inhalation composition according to any one of paragraphs. 1–3, in which the specified API is a macromolecule. 9. Dry powder inhalation composition according to claim 8, containing from 1% wt. up to 85% wt. of said at least one API, relative to the weight of the dry powder inhalation composition. 10. Dry powder inhalation composition according to any one of paragraphs. 1-9, further containing a long-lasting excipient in the lungs, such as a pegylated excipient, or polysaccharides, such as chitosan or dextran. 11. The dry powder inhalation composition according to claim 10, wherein said long-lasting excipient is a PEGylated excipient and is present in an amount ranging from 0.1% by weight. up to 20% by weight, preferably from 0.2 to 10% by weight, more preferably from 0.5 to 5% by weight, based on the total weight of said dry powder inhalation composition. 12. The dry powder inhalation composition of claim 10 or 11, wherein said pegylated excipient is derived from vitamin E or from phospholipids such as tocopherol polyethylene glycol succinate (TPGS) or distearoylphosphoethanolamine polyethylene glycol 2000 (DSPE-mPEG-2000). 13. Dry powder inhalation composition according to any one of paragraphs. 1-12, in which the specified composition is in the form of fine particles having a geometric particle size distribution (PSD) d ₅₀ l, which is less than or equal to 30 μm, preferably less than 15 μm, preferably less than or equal to 10 μm, preferably less than or equal to 5 μm . 14. Dry powder inhalation composition according to any one of paragraphs. 1-12, wherein said composition is in the form of fine particles having a geometric particle size distribution (PSD) d ₉₀ that is less than or equal to 60 µm, preferably less than or equal to 30 µm, preferably less than or equal to 15 µm, preferably less than or equal to 10 µm, and more preferably less than or equal to 7 µm. 15. Dry powder inhalation composition according to any one of paragraphs. 1-12, wherein said composition is in the form of particles having an average volume diameter D[4,3] that is less than or equal to 40 µm, preferably less than or equal to 20 µm, preferably less than or equal to 15 µm, preferably less than or equal to 10 µm, preferably less than or equal to 6 µm. 16. Dry powder inhalation composition according to any one of paragraphs. 1–12, additionally containing an excipient that improves physico-chemical and/or aerodynamic properties, preferably in an amount ranging from 0.1 wt%. up to 80% wt. relative to the total weight of said dry powder inhalation composition. 17. Dry powder inhalation composition according to any one of paragraphs. 1-12 in the form of fine particles having a mass median aerodynamic diameter (MMAD) that is less than or equal to 5 μm, preferably less than or equal to 4 μm. 18. Dry powder inhalation composition according to any one of paragraphs. 1–17 in a blister or capsule for use in a dry powder inhaler or sealed and/or disposable dry powder inhaler. 19. Dry powder inhalation composition according to any one of paragraphs. 1-7 and 10-18, wherein said active agent is a small chemical molecule having bronchodilator activity, glucocorticoid activity, anti-inflammatory activity or anti-infective activity, such as any active pharmaceutical ingredient absorbed by the lung for systemic or local therapy, such as e.g. , budesonide, salbutamol, fluticasone, beclomethasone, mometasone, ciclesonide, formoterol, salbutamol, arformoterol, indacaterol, olodaterol, salmeterol, ipratropium, aclidinium, glycopyrronium, tiotropium, unmeclidinium, mometasone, ciclesonide, formoterol, arformoterol, , tobramycin, vancomycin, tetrahydrolipstatin , clarithromycin, isoniazid, rifampicin, pyrazinamide, itraconazole, voriconazole, aztreonam, ethambutol, streptomycin, kanamycin, amikacin, colistin, sodium colistimethate, capreomycin, ciprofloxacin, rifapentine, doxycycline, cycloserine E, ethionamide, gatifloxacin, cin, moxifloxacin, ofloxacin, fosfomycin , p-aminosalicylate, denufozole tetrasodium, lankovutide, ribavirin, zanamivir, laminavir, rupintrivir, pentamidine, amphotericin B, posaconazole, isavuconazole, capsufungin, micafungin, anidulafangin, iloprost, levothyroxine, their pharmaceutically acceptable salts, solvates, hydrates, polymorphs and their esters, or combinations thereof. 20. Dry powder inhalation composition according to any one of paragraphs. 1-3 and 8-18, in which the specified active agent is a macromolecule, such as a peptide, protein, antibody, antibody fragment, nanobody, nucleic acid, for example insulin, proinsulin, synthetic insulin, semi-synthetic insulin, bevacizumab, pembrolizumab, atezolizumab, nivolumab, ipilimumab, toll-like receptor agonists, ghrelin, IgG monoclonal antibody, small interfering ribonucleic acid (siRNA), dornase alpha, cyclosporine A, antitrypsin alpha-1, interleukin antagonists, interferon-α, interferon-β, interferon-γ, interferon-ω, interleukin-2, monoclonal antibody to IgE, catalase, calcitonin, parathyroid hormone, human growth hormone, insulin-like growth factor I, heparin, rhG-CSF, GM-CSF, Epo-Fc, FSH-Fc, sFc-γ RIIb, mRNA. 21. Dry powder inhalation composition according to any one of paragraphs. 1-18, wherein said active agent is an antitumor agent. 22. The dry powder inhalation composition according to claim 21, wherein said antitumor agent is cisplatin, carboplatin, oxaliplatin, docetaxel, paclitaxel, pemetrexed, etoposide, vinorelbine. 23. The dry powder inhalation composition according to any one of the preceding claims in the form of a dry powder inhalation composition for oral administration.