Dry powder formulation of azole derivative for inhalation

Abstract

A spray dried-powder composition for inhalation comprising particles (X) containing (a) between 5 and 50% by weight of at least one azole derivative in amorphous state but not in crystalline structure and (b) at least one matricial agent to the composition selected from a group consisting of polyol such as sorbitol, mannitol and xylitol; a monosaccharides such as glucose and arabinose; disaccharide such as lactose, maltose, saccharose and dextrose; cholesterol, and any mixture thereof, wherein the composition provides a dissolution rate of said azole derivative of at least, 5% within 10 minutes, 10% within 20 minutes and 40% within 60 minutes when tested in the dissolution apparatus type 2 of the United States Pharmacopoeia at 50 rotation per minute, 37° C. in 900 milliliters of an aqueous dissolution medium adjusted at pH 1.2 and containing 0.3% of sodium laurylsulfate.

Claims

1. Spray-dried particles for an inhalation composition, comprising: a) between 5 and 50% by weight, based on a total dry particle weight of the composition, of at least one azole compound consisting of iconazole, fluconazole, itraconazole, posaconazole, voriconazole, isoconazole, ketoconazole, oxiconazole, bifoconazole, fentoconazole, tioconazole, terconazole, sulconazole, ravuconazole, econazole, and mixtures thereof in an amorphous state; and up to 20 wt. % of the total dry particle weight of at least one azole compound having a nanocrystaline structure with a mean size of between 0.1 and 1 micrometers, wherein said composition attains about 100% dissolution in less than 20 minutes; and b) at least one matricidal compound comprising a polyol comprising sorbitol, mannitol or xylitol; a monosaccharide comprising glucose or arabinose; a disaccharide comprising lactose, maltose, saccharose or dextrose; cholesterol or any mixture thereof.

2. The particles of claim 1, wherein the matricidal compound is mannitol or cholesterol.

3. The particles of claim 1, wherein a weight ratio of the at least one azole compound and at least one matricidal compound is between 0.5/99.5 and 40/60.

4. The particles of claim 1, further comprising a surfactant.

5. The particles of claim 4, comprising between 0.1 and 5% by weight of the surfactant.

6. The particles of claim 4, wherein said surfactant comprises lecithin, phospholipid compounds or hydrogenated phospholipid compounds, or alpha-tocopherol compounds.

7. The particles of claim 6, wherein said phospholipid compounds comprise phosphatic acids, saturated or unsaturated phosphatidyl choline, phosphatidyl ethanol amine, phosphatidyl glycerol, phosphatidyl serine, phosphatidyl inositol, dioleoylphosphatidylcholine, dimyristoyl phosphatidylcholine, dipalmitoyl phosphatidylcholine, distearolyl phosphatidylcholine, dibenolyl phosphatidylcholine, ditricosanoyl phosphatidylcholine, diarachinolyl phosphatidylcholine, dilignocerolyl phosphatidylcholine, dimyristoyl phosphatidyl ethanol amine, dipalmitoyl, phosphatidyl ethanol amine, pipalmitoleoyl phosphatidyl ethanol amine, distearolyl phosphatidyl ethanol amine, dimyristolyl phosphatidyglycerol, or dipalmitoylphosphatidyl glycerol.

8. The particles of claim 1, which provide a Fine Particle Fraction of the azole compound of at least 35% of the total nominal dose of the azole in the particles in accordance with the method “Preparations for Inhalation: Assessment of Fine Particles” using a Multi-Stage Liquid Impinger, Apparatus C, chapter 2.9.18 of the European Pharmacopeia.

9. The particles of claim 5, which contain between about 0.5 and 5% by weight of said surfactant.

10. A method for preparing the spray-dried particles of claim 1, which comprises the steps of: a) preparing a liquid composition, comprising: i) a liquid carrier, comprising a class 3 solvent according to European Pharmacopeia; selected from the group consisting of acetic acid, heptane, acetone, isobutyl acetate, anisole, isopropyl acetate, 1-butanol, methyl acetate, 2-butanol, 3-methyl-1-butanol, butyl acetate, methyl ethyl ketone, tert-butyl methyl ether, methyl isobutyl ketone, cumene, 2-methyl-1-propanol, dimethyl sulfoxide, and pentane; ii) a liquid carrier, selected from the group consisting of ethanol, 1-propanol, ethyl ether, 2-propanol, ethyl formate, propyl acetate, formic acid, mixtures thereof, and any mixtures of the above with water; and iii) at least one azole compound in solution in any of said liquid carriers i) or ii) or both; and iv) at least one matricidal compound in solution in any of said liquid carriers i) or i) or both; and b) spray-drying the liquid composition thereby producing said particles.

11. The method of claim 10, wherein the liquid composition comprises a mixture of class 3 solvents from European Pharmacopeia, or any mixture of two or more of such solvents with or without water.

12. The method of claim 10, wherein said liquid carrier further comprises a surfactant.

13. The method of claim 10, wherein the at least one azole compound is itraconazole.

14. The particles of claim 1, wherein said at least one azole compound is itraconazole.

15. The particles of claim 1, wherein both said amorphous and said nanocrystalline at least one azole compound are itraconazole.

Description

BRIEF DESCRIPTION OF THE FIGURES

(1) FIG. 1 is the MDSC heating curves of spray dried itraconazole.

(2) FIG. 2 is in vitro dissolution profile of micronized crystalline bulk itraconazole, pure amorphous itraconazole and a spray dried powder formulation according to the present invention (example 1B) comprising hydrophilic matricial and itraconazole.

(3) FIG. 3 is in vitro deposition patterns (mean ±S.D, n=3) of spray dried powder formulations according to the present invention (examples 2A to 2D) determined with an MsLI from the Axhaler® device. Results are exposed as percentage of itraconazole (expressed in function of the nominal dose) recovered from the device and each part of the impactor (throat, stage 1, 2, 3, 4 and the filter). The following conditions were used: 100 ml/min, 2.4 s. Three No.3 HPMC capsules filled with a quantity of formulation corresponding to 2.5 mg of itraconazole were used per test.

(4) FIG. 4 is in vitro dissolution profile of bulk crystalline itraconazole and the spray dried formulations according to present invention (examples 2A to 2D).

(5) FIG. 5 is the SEM photographs of spray dried powder formulations according to the present invention (examples 3A to 3E) and a spray dried itraconazole (example 3F) at magnification ×1000.

(6) FIG. 6 is the MDSC heating curves of spray dried powder formulations according to the present invention (examples 3A to 3E), spray dried itraconazole (example 3F) and spray dried mannitol.

(7) FIG. 7 is in vitro deposition patterns (mean ±S.D, n=3) of spray dried powder formulations according to the present invention (examples 3A to 3E) determined with an MsLI from the Axhaler® device. Results are exposed as percentage of itraconazole (expressed in function of the nominal dose) recovered from the device and each part of the impactor (throat, stage 1, 2, 3, 4 and the filter). The following conditions were used: 100 ml/min, 2.4 s. Three No.3 HPMC capsules filled with a quantity of formulation corresponding to 2.5 mg of itraconazole were used per test.

(8) FIG. 8 is in vitro dissolution profile of micronized crystalline bulk itraconazole, spray dried amorphous itraconazole (example 3F) and spray dried powder formulations according to the present invention (examples 3A to 3E).

(9) FIG. 9 is in vitro dissolution profile of spray dried powder formulations according to the present invention (examples 3A to 3E) with Curve A defining the dissolution rate of 5% within 10 minutes, 10% within 20 minutes and 40% within 60 minutes.

(10) FIG. 10 is in vitro dissolution profile of spray dried powder formulations according to the present invention (examples 3A to 3E) with Curves B and B′ defining the dissolution rate of 5% within 5 minutes, 10% within 10 minutes, 15% within 20 minutes and 40% within 60 minutes, and the one of 50% within 5 minutes, 60% within 10 minutes, 90% within 20 minutes and 100% within 60 minutes, respectively.

(11) FIG. 11 is in vitro dissolution profile of micronized crystalline bulk itraconazole and a spray dried powder formulation according to the present invention comprising Itraconazole, cholesterol and phospholipon (example 4).

(12) FIG. 12 is in vitro dissolution profile of micronized crystalline bulk itraconazole and spray dried powder formulations comprising itraconazole and mannitol according to the present invention, i.e., particles not containing crystalline nanoparticles of itraconazole (example 5A) and particles containing crystalline nanoparticles of itraconazole (example 5B).

DESCRIPTION OF THE INVENTION

(13) This invention is related to a dry powder formulation for inhalation of azole derivatives with the proviso that said azole derivative is not a compound of the group consisting of the family of omeprazole, esomeprazole, lansaprazole, pantoprazole and rabeprazole and a process to provide it.

(14) Azole derivatives can be selected from the group consisting of miconazole, fluconazole, itraconazole, posaconazole, voriconazole, isoconazole, ketoconazole, oxiconazole, bifonazole, fenticonazole, tioconazole, terconazole, sulconazole, ravuconazole, econazole, terconazole.

(15) The dry powder of the invention can present high dispersibility capabilities to maximize, after inhalation from an inhaler device, the proportion of particles presenting an appropriated aerodynamic diameter range.

(16) Appropriated aerodynamic range refers to aerodynamic diameter that presents inhaled conidia. Generated particles from an inhaler device in breath conditions must present the same aerodynamic range that inhaled aspergillus conidia (1.9-6 μm) to reach potential infections sites for an optimal treatment targeting and effectiveness.

(17) Advantageously, the dry powder composition is based on the use of exclusively physiological component excipients, safe, generally recognized as save (GRAS) excipients, FDA authorized excipients for inhalation therapy to guaranty a good safety profile after inhalation and to be compatible with the lung membrane to avoid hyper-responsiveness, cough, airway spasticity or inflammation.

(18) The manufacturing process requires one or two step(s) to obtain the final dry product and all techniques used are made for an easy scaling up to industrial batch size production. The dry powder in itself is designed to possess enhanced flow properties for an easy processing at industrial scale.

(19) The dry powder is specifically designed for oral inhalation to treat or give prophylaxis against pulmonary invasive aspergillosis. The azole derivatives are in form that allows that dissolution rate can be improved at different extent and/or modified by varying the composition of the dry powder. The improvement can be controlled by modifying the dry powder composition and/or the active pharmaceutical ingredient (API) physical state or by combining prior administration different embodiments of the invention.

(20) This is advantageous because the modification of dissolution rate can overcome in Vivo clearance and absorption mechanisms that lead to decreasing drug proportion in the site of infection.

(21) The dry powder is constituted of matricial microparticles. The matricial microparticles are constituted of safe, physiological component or inhalation FDA authorized excipient wherein the active ingredient is dispersed in a modified physical state. After inhalation of those microparticles, after matrix dissolution or erosion, the active ingredient will expose a higher surface area to the pulmonary mucosa than the same dose of pure spray dried active ingredient microparticles, resulting in an improved dissolution rate.

(22) The nature of the matricial agent directly influences the dissolution profile of the active ingredient. The matricial agent can be (i) hydrophilic to directly release the active ingredient when in contact with the pulmonary mucosa (ii) hydrophobic to delay the release of the active ingredient (iii) a mixture of hydrophilic and hydrophobic (in different proportion) agent to obtain an intermediate release profile.

(23) Matricial agents are physiological component excipients, GRAS excipients; FDA authorized excipients for inhalation therapy to avoid as far as possible pulmonary or systemic toxicity. The matricial agents can be combined together to confer to the dry powder desired flow, aerodynamic and dissolution characteristics. The matricial agent is necessary in the composition.

(24) Matricial agent can be selected from the group consisting of sugar alcohols, polyols such as sorbitol, mannitol and xylitol, and crystalline sugars, including monosaccharides (glucose, arabinose) and disaccharides (lactose, maltose, saccharose, dextrose) and cholesterol.

(25) In one embodiment of the invention the API is in majority in amorphous state. The proportion of amorphous active ingredient (in percentage of the total amount of active ingredient from the invention is from 51% to 100%, preferably between 70% and 100%, even more preferably 100%.

(26) One way to obtain an amorphous compound is to spray dry it from a solution because the rapid solvent evaporation during the drying process do not let enough time to solid particles to recrystallize. However azole compounds and particularly itraconazole are only sparingly soluble in chloride solvent such as dichloromethane and chloroform which are, due to their high toxicity, not recommended for the preparation of pharmaceutical formulations. This invention provides methods to obtain an amorphous product by spray drying the API from a solution using only a class 3 solvent. Those solvents are considered as low toxic potential solvents and then offer a better safety profile in case of residuals inhalation. This category of solvent includes acetic acid, Heptane, Acetone, Isobutyl acetate, Anisole, Isopropyl acetate,1-Butanol, Methyl acetate, 2-Butanol, 3-Methyl-1-butanol, Butyl acetate, Methylethylketone, tert-Butylmethyl ether, Methylisobutylketone, Cumene, 2-Methyl-1-propanol, Dimethyl sulfoxide, Pentane, Ethanol, 1-Pentanol, Ethyl acetate, 1-Propanol, Ethyl ether, 2-Propanol, Ethyl formate, Propyl acetate, formic acid or the mixture thereof,

(27) By spray drying an organic solution of active ingredient it is possible to obtain it after the drying process in an amorphous state with geometric size appropriated for inhalation therapy (<5 μm). This can be done from a drug saturated organic solution. However solubility of azole derivates such as itraconazole in class 3 solvents is extremely low. These low concentrations could not be optimal for a good recovery of the dry powder after spray drying In order to obtain a good recovery of the dry powder after spray drying azole derivatives with a higher solubility may be selected instead of itraconazole. A matricial agent can be added before spray drying this kind of solutions to enhance total solute concentration. An acid can be added into—a preheated organic class 3 solvent under magnetic stirring in order to enhance the solubility of poorly soluble azole compound such as itraconazole. An organic solution comprising azole compound(s) can also be heated to high temperature under magnetic stirring to obtain enhanced solubility of the azole compound(s). Those options only allow the dissolution of hydrophobic excipients in the solution . . . Λ determinate quantity of water can be added to one of those solutions type in order to allow dissolving both poorly soluble active ingredients, hydrophilic and hydrophobic excipients. This can be particularly interesting in order to modify active ingredient's dissolution rate, particle size, aerodynamic behavior and flow properties. Preferential ratio of water to organic solvent (in volume to volume percentage) are from 0 to 50%, preferably between 0% to 30%, more preferably between 10% and 30% and even more preferably between 20% and 30%.

(28) On a thermodynamic point of view, due to their unorganized structure, amorphous compounds present the advantage to possess higher solubility than the same crystalline compound. In practice, during dissolution, amorphous compounds often recrystallize to lower energy crystalline state presenting lower solubility than the initial product. This invention provides formulations wherein an active compound is in an amorphous state and formulated so that its dissolution occurs before complete drug recrystallization leading to an improved dissolution rate product. Indeed, the improvements and enlargement of surface area of dry powder formulation arrived at local site of a patient can be obtained by spray drying a solution of an active ingredient together with a hydrophilic matricial agent which provides particles comprising the active ingredient in amorphous state dispersing in the matricial agent. Such improvements in surface area can—accelerate the active ingredient dissolution rate preventing from excessive recrystallization prior dissolution.

(29) Recrystallization of amorphous drugs also may happen during storage leading to a decrease of the dissolution performance product. One aspect of the present invention provide a stable amorphous product when formulate as a solid dispersion of the active ingredient in a matricial agent.

(30) In a composition of the invention, the amount of azole derivates that can be incorporated in the matricial agent(s) is from 0.5 to 40%, preferably from 1 to 35%, more preferably from 10 to 35% by weight.

(31) Surprisingly, it is possible by varying the concentration of the spray dried solution or the matricial agent/API ratio to modify aerodynamic behavior of generated particles. Varying the concentration in solution or the matricial agent/API ratio can directly modify the geometric diameter and the density of dried particles thus their aerodynamic diameter which will also directly modify their aerodynamic behavior. Modifying one of those parameters would lead to formation of particles presenting different aerodynamic behavior while presenting similar dissolution rate. This can help to provide a dry powder with an optimized dissolution rate that will penetrate the lung in a sufficient quantity to provide appropriated antifungal dose from a predetermined nominal dose. Variation of those parameters allows then the optimization of the fine particle dose (FPD) of the spray dried powder while keeping improved dissolution rate.

(32) Preferably, the amount of the azole derivative added in the liquid composition is between 0.1% and 5%, preferably between 0.5% and 2% by weight of the azole derivative to the volume of the liquid composition (g/100 mL).

(33) A surfactant can be added in the matrix of particles comprised in a dry powder formulation according to the present invention in order to improve the dissolution rate enhancement of the active ingredient. A surfactant is an amphiphilic compound with both hydrophilic and hydrophobic characteristics. By spray drying a solution containing both the active ingredient the matricial agent and a surfactant it is possible to produce matricial microparticles wherein the active ingredient and the surfactant are dispersed. The surfactant will play a wetting enhancement effect on the active ingredient resulting, in a reduction in particle agglomeration and acceleration/improvement of its dissolution rate when compared to matricial microparticles without surfactant.

(34) The surfactant(s) can be selected from the group consisting of physiological component, GRAS (generally recognized as save) excipients, FDA authorized excipients for inhalation therapy to avoid any pulmonary or systemic toxicity.

(35) The quantity of added surfactant could influence azole compound dissolution rate improvement. The preferred amount of surfactant is comprised between 0.1 and 5% by weight in the dry powder composition.

(36) Preferentially surfactant can be phospholipids, lecithin, lipids or GRAS modified vitamins, or combination of such surfactant. Phospholipids that may use comprise phosphatic acids, phosphatidyl choline (saturated and unsaturated), phosphatidyl ethanol amine, phosphatidyl glycerol, phosphatidyl serine, phosphatidyl inositol. Examples of such phospholipids include, dioleoylphosphatidylcholine, dimyristoyl phosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), diarachidoyl phoshatidylcholine (DAPC), dibenoyl phosphatidylcholine (DBPC), ditricosanoyl phosphatidylcholine (DTPC), dilignoceroylphatidylcholine (DLPC), dimiristoylphosphatidylethanolamine (DMPE), dipalmitoylphosphatidylethanoalamine (DPPE), pipalmitoleoylphasphatidylethanol amine, distearoylphosphatidylethanolamine (DSPE), dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidyl glycerol (DPPG), dipalmitolcoylphosphatidylglycerol and more preferentially hydrogenated derivates. Examples of GRAS modified vitamins comprise α-tochopherols derivates.

(37) A too high quantity of surfactant in the formulation can induce an important particle size increase during spray drying. Due to their low melting point, surfactants could soft or melt during spray drying increasing particle size. Dilution of the surfactant in the matricial agent can mask this effect resulting in production of smaller particles with appropriate characteristics.

(38) One particular embodiment of the invention consists to obtain the active ingredient in the form of crystalline nanoparticles by a method described in the art.

(39) The term “nanoparticles” used to describe the present invention has a meaning of solid discrete particles ranging in size from 1 nm to 1000 nm. The presence of the crystalline nanoparticles of azole derivative in a spray dried particle and the weight ratio of the crystalline nanoparticles comprised in the particle can be determined by using powder X-ray diffraction, and differential scanning calorimetry concomitantly with IIPLC drug quantification.

(40) Those nanoparticles are then dispersed in a matricial agent to confer to the formulation appropriated particle size, flow properties, dissolution rate and aerodynamic behavior. The dissolution rate of those nanoparticles is instantaneous (within 5 minutes) with a very pronounced burst effect that cannot be delayed due to inherent dissolution rate of the nanoparticles.

(41) The production of this formulation types (i.e., particles containing crystalline nanoparticles of the active ingredient and the matricial agent) includes two steps in the manufacturing procedure. The first step being the production of drug nanoparticles and the second step being the drying procedure. The nanoparticles could be produced by a method described in the art. Preferably nanoparticles are produced by high pressure homogenization. The matricial agent can be added prior the size reduction step or before the spray drying procedure.

(42) In one particular embodiment of the invention the active ingredient is dispersed in the matricial agent both in form of crystalline nanoparticles and amorphous compound. This embodiment can be the result of the spray drying of both matricial agent and the active ingredient in solution together with nanoparticles of the active in. Another aspect of this embodiment is that the dry powder formulation according to the present invention is manufactured by a simple blend of the nanoparticles of the active ingredient, which are obtained by spray drying of a suspension comprising its crystalline nanoparticles and a matricial agent or by mechanical milling of the crystalline active ingredient, and an amorphous matricial formulation obtained by spray drying of the active ingredient in solution. This blend powder will be filled in capsule, blister or multidose device.

(43) The desired result is to confer to the formulation a controlled dissolution profile by optimizing the proportion of nanoparticles/amorphous compound in the formulation. This dissolution profile could not be reach with only the nanoparticles in the formulations. The modification of the proportion nanoparticles/amorphous allow varying dissolution profile. Preferably, the ratio (w/w) of amorphous matricial particles/nanocrystalline matricial composition is comprised between 100/0 to 80/20.

(44) In another embodiment the active ingredient is dispersed as nanoparticles or microparticles in a matrix of the same active ingredient. The active ingredient matricial being in amorphous state

(45) Nanosuspension could be concomitantly spray dried with a solution of active ingredient containing a matrix former. The differences that exist between amorphous and nanoparticles dissolution rate could allow modifying dissolution rate of the formulation. The API in solution could either be used as matrix former encapsulating the nanoparticles. This could provide formulation presenting an interesting dissolution rate and optimal aerodynamic characteristics.

EXAMPLES

Example 1

(46) The starting material is constituted of crystalline micronized itraconazole (ITZ) with a volume mean diameter of 3.5 μm and 90% of particles below 6.2 μm. Pure amorphous itraconazole (Example 1A) and a hydrophilic matricial formulation of itraconazole dry powder (Example 1B; invention) were produced at laboratory scale by spray-drying using a Büchi Mini Spray Dryer B-191a (Büchi laboratory-Techniques, Switzerland). Two feed stock solutions were prepared then separately spray-dried in the following conditions: spraying air flow, 800 l/h; drying air flow, 35 m.sup.3/h; solution feed rate, 2.7 g/min; nozzle size, 0.5 mm; Inlet temperature, 90° C.; resulting outlet temperature of 53° C. The composition of the feedstock solutions is summarized in Table 1. Each component were dissolved under magnetic stirring (600 rpm) in a hydro-alcoholic solution (20 water-80 isopropanol) heated at 70° c. During spray drying the solutions were kept at a temperature between 60 and 70° C.

(47) TABLE-US-00001 TABLE 1 Composition of spray dried solutions in Example 1. liquid Itraconazole Mannitol Isopropanol Water composition (g) (g) (ml) (ml) Example 1A 0.56 — 80 20 (Comparative: Cex) Example 1B 0.56 1 80 20 (Invention: INV)

(48) Crystallinity profile of the dried samples was evaluated using MDSC (modulate temperature differential scanning calorimetry) and PXRD (powder x-ray diffraction). Those two techniques are complementary and provide a maximum of information on sample's polymorphism.

(49) MDSC experiments were conducted using a Q 2000 DSC (TA Instruments) equipped with cooling system. MDSC differs from standard DSC in the possibility to apply two simultaneous heating rates to the sample, a sinusoidal modulation is added to the linear heating ramp. The total measured heat flow corresponds to the standard heat flow in classic DSC. MDSC heating conditions offers the possibility to make the deconvolution of reversing and non reversing heat flow in which particular thermal event can be singularly detected. Crystallizations phenomena were then observed in the non-reversing heat flow, glass transitions were observed in the reversing heat flow while melting were observed in total heat flow All samples were analyzed in the same following conditions. A 2-3 mg sample was exactly weighted in a low mass aluminum hermetic pan. A 5° C./min temperature rate with a modulation of +/−0.8° C. every 60 seconds was applied to the sample from 25° C. to 185° C. The instrument was calibrated for temperature using indium as a standard. The heat flow and heat capacity signals were calibrated using a standard sapphire sample. The Universal Analysis 2000 software was used to integrate each thermal event.

(50) PXRD is a powerful tool widely used to evaluate the crystalline form of various compounds. It can help to determine the structural physical state of a product. At a given crystalline lattice, will correspond a given PXRD spectra and inversely a given chaotic system (as amorphous state) would not provide any diffraction peak. This will therefore help to evaluate the polymorphic form obtained after spray drying and in a second time to estimate the proportion of amorphous phase within a sample. The powders were analyzed by the Debye-Scherrer method. The samples were submitted to the Kα line of copper, monochromatic radiation (λ=1.540 Å). The diffractometer (Siemens D5000, Germany) equipped with a mounting said reflection Bragg-Brentano, connected to the monochromator and a channel program Diffracplus. The measures are determined to 40 KV, 40 mA in 2theta an angular range from 2° to 60° in steps of 0.02° through a counting speed of 1.2 s per step and a rotation speed of ‘sample of 15 rpm. Each sample was stored in a hermetic plastic container and placed at 8, 25, 40° C. They were analyzed directly after spray drying, and after 2 months storage at the different temperatures.

(51) It is possible to quantify the percentage of crystalline phase in a given compound. Several techniques of calculation have been developed In this case measuring the areas under the curves was used to determine the percentage of amorphous phase in the sample. Indeed, there is a proportional relationship between the ratio of the area under the curve of the diffraction peaks above the deviation from the baseline (A.sub.c) and the total area of the diffractogram (A.sub.tot) with the amount of crystalline phase in the sample. To calculate the degree of crystallinity within a sample it suffices to measure the area under the curve of the diffraction peaks (A.sub.c) without integrating the deviation from the baseline because it comes from the noise and amorphous areas present in the sample. Then integrate the total area under the curve of the diffractogram (A.sub.T). The percentage of crystalline phase will be expressed as in equation 1. The amorphous content expressed in % was estimated as 100% minus the estimated crystallinity degree.

(52) $\begin{array}{cc}\%\mathrm{Crystailinity}=\left(\frac{A_C}{A_T}\right)\times 100& \mathrm{Equation}1\end{array}$

(53) MDSC analysis (FIG. 1) showed that amorphous itraconazole (Example 1A) exhibited a glass transition at about 49° C.

(54) An exothermic recrystallization peak was observed between 100° C. and 125° C., which was followed by an endothermic peak around 164° C. that corresponded to the melting of early formed crystalline material. This crystalline itraconazole melted at a temperature lower that the bulk material when analyzed in the same conditions (about 168° C.). Those thermal events are characteristics of glassy itraconazole.

(55) PDRX confirmed amorphous state of itraconazole in Examples 1A and 1B. At T 0 month no diffraction's peak appeared on diffractogram of Example 1A. Approximated calculated amorphous phase in this sample was equal to 100%. This traduced the lack of any crystalline structure in the sample.

(56) TABLE-US-00002 TABLE 2 DRX based estimated amorphous sample's content Formulation T 0 months T 2 months 8° C. Example 1A (Cex) 100% 100% Example 1B (INV) 52% 52% 25° C. Example 1A (Cex) 100% 100% Example 1B (INV) 52% 55% 40° C. Example 1A (Cex) 100% 63% Example 1B (INV) 52% 55%

(57) No recrystallization occurred after 2 months of storage at 8, and 25° C. The percentage of amorphous phase stayed at 100% and no diffraction's peak characteristics of crystalline itraconazole were observed in the diffractograms. When stored at 40° C. amorphous itraconazole recrystallized and approximated amorphous phase shifted to 63%. Recrystallization peaks appeared at the originals diffraction's angles of bulk crystalline itraconazole signifying that amorphous itraconazole recrystallized to its original more stable form.

(58) At T 0, Example 1B's diffractogram exhibited some diffractions peaks. However none of those peaks corresponded to crystalline itraconazole. Diffraction profiles of both α, β and δ mannitol were present. Total approximated amount of amorphous phase within the sample was equal to 52%. This value was higher than actual content of itraconazole in the sample. This came probably from the proportion of mannitol that was amorphous after spray drying. When stored at 8° C., 25° C. and 40° C. only small variations in the approximated amorphous phase in the sample was observed (see Table 2). Contrary to Example 1A, no recrystallization evidences of itraconazole were present at its characteristics diffractions angles. Dispersing amorphous itraconazole in mannitol (by spray drying a solution containing both components) yielded to the stabilization of the amorphous API.

(59) Aerodynamic behavior of generated particles after dose actuation from a dry powder inhaler was assed using a multistage liquid impinger (MsLI). The dry powder inhaler used was an Axahaler® (SMB laboratories). A flow rate (adjusted to a pressure drop of 4 kPa) of 100 L/min during 2.4 sec was applied through the device for each actuation. The device was filled with HPMC n°3 capsules loaded with an approximate quantity of dry powder corresponding to 2.5 mg of itraconazole. One test was realized with three discharges. After the three dose actuations the total deposited dry powder was quantified for each part of the impactor with a suitable and validated HPLC method. Each test was replicated three times. For each test the fine particle dose (FPD) has been estimated by the method described in the European Pharmacopeia 7.2 for aerodynamic assessment of fine particle, apparatus C (MsLI). The expressed results have been weighted to a constant itraconazole nominal dose of 2.5 mg. The fine particle fraction (FPF) is the FPD expressed in % of the nominal dose.

(60) A Malvern Spraytec® laser diffraction equipment was used to measure particle size distribution (PSD) during the aerodynamic fine particle assessment test. The laser beam was directly placed between the throat and the impactor to measure the PSD of generated dry powder cloud, which was then split along its aerodynamic diameter in the MsLI during simulated inhalation conditions. The average PSD was measured from three replicates of each sample. Results were expressed in terms of D[4.3], d(0.5) and d(0.9) which are, respectively, the volume mean diameter and the size in microns at which 50% and 90% of the particles are smaller than the rest of the distribution. Results are expressed in Table 3.

(61) TABLE-US-00003 TABLE 3 Size and aerodynamic characteristics of the different formulations: Particle Size Characteristics (Mean ± SD, n = 3) Measured with the the Spraytec ® and fine particles fractions (% of particle with d.sub.ac < 5 μm) expressed in function of nominal dose (FPF; Mean ± SD, n = 3) measured by impaction test (MsLI). Spraytec ® MsLI Formulation d(0.5) (μm) D[4.3] (μm) d(0.9) (μm) FPF (%) Example 1B 2.22 ± 0.11 2.75 ± 0.39 3.38 ± 0.28 46.9 ± 1.9 (INV)

(62) Particle size analysis revealed that the volume mean diameter of the invention was below 5 μm which is the first criteria for deep lung deposition. This was confirmed by the aerodynamic fine particle assessment test. The invention presented a high FPF equal to 46.9±1.9%.

(63) Dissolution tests were performed using USP 33 type 2 paddle apparatus (Distek Dissolution System 2100C, Distek Inc., USA). The dissolution media was constituted of desionized water set at pH 1.2 (HCl 0.063N) containing 0.3% of sodium lauryl sulfate. This dissolution allowed maintaining SINK conditions throughout the test. The medium was heated to 37° C. and kept at this temperature during the test. The paddle speed was set at 50 rpm and the dissolution vessel was filled with 900 ml of dissolution media. An exactly weighted amount of dry powder corresponding to 10 mg of itraconazole was spread on the dissolution media (=T0). Itraconazole was quantified at pre-determined intervals (0, 2, 5, 10, 20, 30, 60, and 120 minutes) using a suitable validated HPLC method. Five milliliters of dissolution media was removed from the dissolution vessel and directly replaced by fresh dissolution medium. These five milliliters were directly filtered through 0.2 μm diameter filters to avoid quantification of undissolved particles at the determinate time interval. The cumulative amount of drug release was calculated and expressed in percentage of initial drug load and plotted versus time. Each test was replicated three times.

(64) Dissolution profiles are shown in FIG. 2. Comparison of the dissolution curves of crystalline micronized (bulk ITZ) and pure amorphous ITZ (Example 1A) suggested no difference in the drug release curves. This observation was interesting, since amorphous ITZ would be expected to have a faster dissolution profile compared to the crystalline ITZ. This may come from the fact that the highly hydrophobic nature of the drug substance could lead to poor wetability by the aqueous dissolution media impeding drug dissolution improvement.

(65) Progressive re-crystallization of amorphous ITZ could also have occurred during dissolution, delaying dissolution of the amorphous form. However, it was surprisingly discovered that the formulation of Example 1B according to the present invention wherein ITZ is dispersed in mannitol microparticles provided a significant improvement of the dissolution rate of ITZ, i.e., 11.4% at 10 min, 15.2% at 20 min and 46.7% at 60 min, compared to bulk micronized crystalline ITZ and pure amorphous ITZ. The increase in surface area available to the dissolution media of amorphous ITZ, when dispersed in mannitol microparticles could explain this significant acceleration (FIG. 2) of dissolution rate. Mannitol being dissolved quasi instantly, it was supposed that remaining ITZ particles exposed a higher surface area to the dissolution media that pure spray dried amorphous particles. Mannitol formed spherical matrix wherein amorphous ITZ is dispersed. Once the mannitol is dissolved, porous amorphous ITZ particles are released in the dissolution vessel whit, due to numerous pores formed by the mannitol dissolution. The increased surface area available to the dissolution media increases dissolution rate and prevents excessive re-crystallization which enhance solubility therefore dissolution rate.

Example 2

(66) The purpose of this example was to demonstrate the ability of the invention to modify aerodynamic behavior of the dry powder without modifying its dissolution rate by modifying excipient/API ratio and the total solute in the liquid composition for spray drying.

(67) Four formulations were prepared at laboratory scale by spray-drying using a Büchi Mini Spray Dryer B-191a (Büchi laboratory-Techniques, Switzerland). Four feed stock solutions were separately prepared and spray dried. A determined quantity of itraconazole and mannitol (see Table 4) were dissolved in 100 ml of a hydro-alcoholic solution (20 water-80 isopropanol) heated at 70° C. under magnetic stiffing (600 rpm). The total dry product amount in solution for Examples 2A and 2B are similar (1.56 g). The only difference between the two formulations is the ratio of itraconazole/mannitol. For formulation 2A, 2C and 2D the ratio of itraconazole/mannitol was constant but the total amount of solute in solution in the liquid composition was different. The spray drying conditions are the same that in Example 1.

(68) TABLE-US-00004 TABLE 4 Amount of itraconazole and mannitol in the liquid compositions for spray drying in Example 2 Liquid composition Composition (for 100 ml) Example 2A (INV) Itraconazole 0.56 g Mannitol 1 g Example 2B (INV) Itraconazole 0.234 g Mannitol 1.326 g Example 2C (INV) Itraconazole 0.28 g Mannitol 0.5 g Example 2D (INV) Itraconazole 0.84 g Mannitol 1.5 g

(69) Crystallinity profile of samples was assed using PXRD (powder x-ray diffraction) at the same condition that those described in Example 1.

(70) The diffractograms of the four formulation presented some diffraction's peaks. However none of those diffraction's peaks corresponded to crystalline itraconaozole. That means that itraconazole, in those formulations, was in an amorphous state. Mannitol was in majority in crystalline state. Its three different polymorphic forms (α, β and δ) were present in all samples but in different proportions, the δ form being in majority.

(71) Powder flowability was evaluated by Carr's compressibility index (CI) as described in Example 1. A Carr's index values of above 40% are generally related to poor powder flowability whereas value under 20% are related to extremely good powder flowability. The four present a CI value ranging from 20.9% to 28.8%. Those values indicate good powder flowability for both formulations.

(72) Particle size distribution of powders was evaluated by laser scattering using a Malvern Mastersizer 2000® (Malvern instrument) via a Sirocco 2000® (Malvern instrument) dry feeder dispersion unit. Particle size measurement was done on a sample of +/−50 mg at a pressure of 4 Bar with a feed rate vibration set at 40%. Those conditions allow to measure particle size distribution of practically, totally desagglomerated powder due to very drastic dispersion conditions. Particle refractive index with a real part equaling 1.48 and imaginary part of 0.1 were chosen. Those values ensure low weighted residual (<2%) which traduces result's integrity.

(73) A Malvern Spraytec® was used as describe in example 1. For both techniques, the average PSDs was measured from three replicates of each sample. Results were expressed in terms of D[4.3], d(0.5) and d(0.9) which are, respectively, the volume mean diameter and the size in microns at which 50% and 90% of the particles are smaller than the rest of the distribution. Results are expressed in Table 5.

(74) Aerodynamic behavior of generated particles was evaluated by impaction test as described in Example 1. The fine particle fraction is the FPD expressed in % of the nominal dose (FPF) having an aerodynamic diameter inferior to 5 μm. The emitted doses have been calculated and correspond to the recovered dose from the induction port and five stages of the MsLI during the tests. The emitted dose is express in percentage of the nominal dose and corresponds to the percent of the nominal dose that effectively leaved the device and capsule. Results are expressed in Table 6 and represented in FIG. 3.

(75) Malvern Sirocco® measurements showed that the four formulations exhibited similar mass median diameter d(0.5), and the volume mean diameter values (D[4.3]) of the formulations 2B and 2C were higher than those of the two other formulations as expressed in Table 5. The formation of slightly larger particles seemed occurred in those two formulations. In addition their deagglomeration seemed to be more difficult regarding higher d(0.5) and D[4.3] values obtained for the 2B and 2C formulations with Spraytec® analysis in simulated breath conditions.

(76) TABLE-US-00005 TABLE 5 Size characteristics of the different formulations of Example 2: Particle Size Characteristics (Mean ± SD, n = 3) were Measured with the Malvern Masterzizer2000 ® and Spraytec ® Malvern Sirocco ® Spraytec ® d(0.5) D[4.3] d(0.9) d(0.5) D[4.3] d(0.9) Formulation (μm) μm (μm) (μm) (μm) (μm) Example 2A (INV) 0.74 ± 0.01 1.00 ± 0.04 1.78 ± 0.09 2.22 ± 0.11 2.75 ± 0.39 3.38 ± 0.28 Example 2B (INV) 0.73 ± 0.03  1.2 ± 0.46 1.89 ± 0.49 2.99 ± 0.11 6.45 ± 1.78 14.91 ± 9.94  Example 2C (INV) 0.76 ± 0.03 1.54 ± 0.18 3.08 ± 0.75 2.70 ± .05  4.60 ± 0.62 7.12 ± 2.20 Example 2D (INV) 0.76 ± 0.01 1.01 ± 0.04 1.86 ± 0.12 2.16 ± 0.04 2.31 ± 0.04 2.90 ± 0.03

(77) Despite their higher particle size and their lower deagglomeration efficiency, the 2B and 2C formulations have higher FPF than formulations 2A and 2D. This is directly related to higher emitted dose for those two formulations (2B and 2C). Because of extremely fine granulometry, despite lower deagglomeration tendency and slightly larger particle size those two formulations penetrated deeper in the impactor than formulation 2A and 2D which result in higher FPF.

(78) TABLE-US-00006 TABLE 6 Particle deposition, FPD and FPF (mean ± SD) and emitted dose (% nominal dose) obtained during impaction test (MSLI, 100 l/min, 2.4 sec, 3 discharges per test, nominal dose weighted at 2.5 mg, n = 3). Example 2A Example 2B Example 2C Example 2D Mean FPD 1.17 ± 0.05 1.40 ± 0.01  1.36 ± 0.09 1.19 ± 0.04 (mg) Mean FPF (%) 49.6 ± 1.9 56 ± 0.4 54.4 ± 1.8 47.6 ± 1.6 Emitted 53.3 ± 1.9 71 ± 0.5 73.5 ± 6.3 53.3 ± 1.5 dose.sub.nom (%)

(79) Dissolution tests were conducted as described in Example 1. Obtained dissolution profiles are shown in FIG. 4. The four formulations exhibited different and faster dissolution's rate than bulk micronized crystalline itraconazole (FIG. 4). The dissolution profiles of Examples 2A, 2B, 2C and 2D were similar.

(80) Regarding those results it is possible to modify aerodynamic behavior of generated particles by modifying active ingredient/matrix former ratio, the total amount of solute or the concentration of the active ingredient in solution of the spray dried solution while keeping similar dissolution profile. The modification of the aerodynamic behavior was done without varying excipient type or spray drying parameters. This shows the possibility of this flexible one step process to vary aerodynamic behavior of particles without modify API dissolution rate. All excipients used were GRAS. The four formulations presented good powder flowability.

Example 3

(81) The purpose of this example was to show the ability of the invention to modify dissolution rate's acceleration of a formulation while keeping good flow properties and aerodynamic characteristics.

(82) Three formulations were produced at laboratory scale by spray drying feed stock solutions using a Büchi Mini Spray Dryer B-191a (Büchi laboratory-Techniques, Switzerland). For the five examples a determined quantity of itraconazole, mannitol and hydrogenated soy-lecithin with more than 90% of hydrogenated phosphatidylcholine (Phospholipon 90H), (see Table 7) were dissolved in 100 ml of an hydro-alcoholic solution (20 water:80 isopropanol) heated at 70° c under magnetic stirring (600 rpm). The spray drying conditions are the same that in Example 1.

(83) TABLE-US-00007 TABLE 7 Theoretical composition of spray dried solutions, dry formulations ns used during the spray drying process in Example 3. Liquid composition Dry powder composition ITZ % Mannitol PL90H % ITZ Mannitol PL90H Formulation (w/v) %(w/v) (m/m.sub.ITZ) (% w/w) (% w/w) (% w/w) Example 3A (INV) 0.56 1 — 35.9 64.1 — Example 3B (INV) 0.1 0.9 — 10 90 — Example 3C (INV) 0.56 1  1 35.77 63.87 0.36 Example 3D (INV) 0.56 1 10 34.65 61.88 3.47 Example 3E (INV) 0.1 0.9 10 9.90 89.11 0.99 Example 3F (Cex) 0.56 — — 100 — —

(84) Determination of drug content was used in order to compare expected and actual drug content. For that a determined quantity of dry powder was dissolved in a dilution phase and sonicated during 20 min Those solutions were analyzed by HPLC-UV from which the drug content (wt %) was determined. Average content (wt %) and standard deviations were calculated from five analysis. Itraconazole content measurements results for the different formulations are summarized in Table 8. The measured values were very close to the expected one with relative errors ranged between −3.9% and 3.0%. Lower itraconazole content as well as introduction of phospholipids in the formulations induced a reduction of this relative error. The active ingredient seemed to be uniformly distributed within particles since samples have been selected randomly and that variation coefficient for all five test samples were not greater than 3.25%. Those exact ITZ contents values were used during aerodynamic particle size analysis to determine exact nominal doses. No ITZ degradation seemed to occur during the spray drying process. The relative error between the measured and expected ITZ content for pure spray dried itraconazole (Example 3F) was equal to 0.7%.

(85) TABLE-US-00008 TABLE 8 ITZ content measured by HPLC determination of spray dried powder of Example 3 (mean +/− SD; n = 5) Coefficent Expected Relative Measured ITZ variation ITZ content error Formulation content (wt %) (%) (wt %) (%) Example 3A (INV) 34.5 ± 0.6 1.64 35.9 −3.9 Example 3B (INV) 9.99 ± 0.3 3.25 10 −0.1 Example 3C (INV) 35.6 ± 0.7 1.82 35.8 −0.6 Example 3D (INV) 33.6 ± 0.7 2.01 34.65 −3.1 Example 3E (INV) 10.2 ± 0.2 1.98 9.9 3.0 Example 3F (Cex) 100.7 ± 1.6  1.61 100 0.7

(86) Qualitative morphological evaluations were conducted by scanning electron microscopy using a Philips XL30 ESEM-FEG (FEI, The Netherlands). The samples were spread on a carbon adhesive band then coated with gold at 40 mA for 90 sec at 6.10-2 mbar under argon. Observations were done at acceleration between 5 and 25KV depending on the sample.

(87) Regarding the quantitative composition of the spray dried formulations, mannitol was the major component and was therefore subject to forming matricial particles within which were dispersed the ITZ and, if applicable, the PL. The morphological evaluation showed that very small spherical particles (˜1-2 μm with presence of submicron size particles) with smooth surfaces were formed from the spray dried solution containing mannitol and itraconazole without PL (Examples 3A and 3B; FIG. 5). No morphological differences were observed between these formulations despite the different proportions of amorphous content and mannitol polymorphs. However Example 3B seems to be constituted of slightly larger spherical particles. The presence of PL induces the formation of larger particles with a granular appearance. For the formulation presenting the highest PL content (Example 3D), this granular appearance was the more pronounced and interparticular links were observed (FIG. 5). Those links were probably formed during the spray drying process because of the softening or melting of PL inducing particle aggregation. The reduction of the PL content (Examples 3C and 3E) considerably reduced this grainy aspect.

(88) The residual moisture and solvent content of the different dry powders was assessed using thermogravimetric analysis (TGA) with a Q500 apparatus (TA instruments, New Castle, USA) and Universal Analysis 2000 version 4.4A software (TA Instruments, Zellik, Belgium). The residual water and solvent content was calculated as the weight loss between 25° C. and 125° C. and expressed as a percentage of the initial sample mass. Run were set from 25° C. to 300° C. at a heating rate of 10° C./min on sample mass of about 10 mg and performed in triplicate. Weight loss measured during heating the samples between 25° C. and 125° C. were very low (<0.5%) for each formulations.

(89) MDSC were realized as described in Example 1 and results are show in FIG. 6. As previously described (Example 1A), spray dried in those conditions MTDSC analysis showed that itraconazole was retrieved in its particular amorphous glassy state after the spray drying process (here Example 3F). This particular profile was also observed again on MDSC thermograms for the formulations containing the highest proportion of itraconazole (Examples 3A, 3C and 3D; FIG. 6). The glass transition at about 49° C. was present in the reversing heat flow as was the cold crystallization exotherm at around 100° C. in the non-reversing heat flow. Those thermal events were not detected in the formulations containing the smallest proportion of itraconazole (˜10%; Examples 3B and 3E), probably due to lack of sensitivity of the thermal detection for diluted compositions. Spray-dried mannitol and itraconazole melted (total heat flow) at around the same temperature. One single melting point at about 164° C. was observed for all formulations. One supplementary endothermic peak followed by an exothermic peak around 150° C. was observed for Example 3E. These transitions correspond to the melting of δ-mannitol followed by crystallization into the β polymorph. The other formulations did not exhibit this thermal event probably because mannitol was only almost totally (98.5%) in the δ form in this formulation (see PXRD results).

(90) PXRD analyses were conducted on all spray dried powders as described in Example 1. Amorphous contents calculated using the area under the diffractograms are summarized in Table 9. Formulation with higher ITZ content exhibited higher amorphous content. A good correlation was obtained between calculated amorphous content and itraconazole measured content by HPLC (R.sup.2>0.9).

(91) the proportion of participation of each mannitol polymorph to the formation of the total crystalline network was evaluated using the reference intensity ratio methodology. Calculations were made on Diffracplus EVA software. This semi-quantitative method of estimation consists of the identification of the different phases in a specimen by comparison with references patterns (from ICDD data base) and the relative estimation of the proportions of the different phases in the multiphase specimens by comparing peak intensities attributed to the identified phases.

(92) TABLE-US-00009 TABLE 9 PXRD based estimated amorphous content and α β and Δ mannitol β Δ Amorphous α mannitol mannitol mannitol Formulation content (%) (%) (%) (%) Example 3A (INV) 55 42 2.5 55.1 Example 3B (INV) 34.5 38.3 1.3 60.3 Example 3C (INV) 53 31.4 2.9 65.7 Example 3D (INV) 57 1.1 0.4 98.5 Example 3E (INV) 37 20.5 0.9 78.8 Example 3F (Cex) 100 — — —

(93) One specific diffraction peak was chosen for each polymorph were no other crystalline structure that could be present in the dry powder diffracted. Specific diffraction peaks at 43.92, 16.81 and 22.09° 2θ were used for α, β and δ-mannitol, respectively and their respective ICDD spectrum were adjusted to those diffractions ray for calculation. The results are expressed as an estimation of the percentage of each polymorph in the formulations and are summarized in Table 9.

(94) Flow properties were evaluated by determining the Carr's index compressibility index (CI) as described in Example 2. Good powder flowability is a necessary characteristic for an eventually easy processing at an industrial scale. Moreover, more specifically to dry powder for inhalation, a good flowability has already been related to generate an adequate metering, dispersion and fluidization of a dry powder from an inhaler device. All formulations exhibited CI values ranged between 15.6% and 26.4% (see Table 10) which indicated good potential in flow properties for this formulations type.

(95) Particle size analyses were conducted using two different methods. The first method (using a Malvern Mastersizer2000®) provided size results corresponding to totally individualized particles. The second method (using a Malvern Spraytec®) allowed evaluating the size of particles in a deagglomeration rate that is produced after dispersion form an inhaler device.

(96) Malvern Mastersizer2000® results showed that all formulations presented a very fine granulometry with a volume mean diameter ranged from 1.00 μm to 2.04 μm and a mass volume median diameter comprised between 0.74 μm and 1.81 μm (Table 10). The PSD of formulations without PL, Examples 3A and 3B, were very close with a d(0.5) value of 0.74 μm and 0.88 μm, respectively. However, as observed by SEM a small proportion of larger particles were formed for Example 3B, which was traduced by an increase in the D[4.3] and d(0.5).

(97) TABLE-US-00010 TABLE 10 Size, aerodynamic and flow characteristics of formulations obtained fromt he different solutions: Particle Size Characteristics (Mean ± SD, n = 3) Measured with the Mastersizer 20000 ® and the Spraytec ®, Emitted dose (expressed in % of nominal dose) and fine particles fractions (% of particle with dae < 5 μm) measured by impaction test (Mean ± SD, n = 3, Carr's index value (CI) (Mean ± SD, n = 3). Laser light scattering Mastersizer 2000 ® Spraytec ® Aerodynamic evaluation Formu- d(0.5) D[4.3] d(0.5) D[4.3] ED FPF Cl lation (μm) (μm) (μm) (μm) (%.sub.nom) (%.sub.nom) (%) Ex. 3A 0.74 ± 0.01 1.00 ± 0.04 2.2 ± 0.1 2.8 ± 0.4 53.3 ± 1.9 46.9 ± 1.9 26.4 ± 0.1 Ex. 3B 0.88 ± 0.07 1.15 ± 0.05 2.71 ± 0.08 3.66 ± 0.07 81.9 ± 0.6 67.0 ± 1.0 20.6 ± 0.8 Ex. 3C 1.35 ± 0.01 1.59 ± 0.01 2.97 ± 0.04 3.14 ± 0.08 68.3 ± 7.8 52.5 ± 4.9 18.1 ± 2.1 Ex. 3D 1.81 ± 0.05 2.04 ± 0.05 4.63 ± 0.01 5.27 ± 0.07 75.2 ± 4.6 43.0 ± 5.2 24.9 ± 0.9 Ex. 3E 0.93 ± 0.01 1.23 ± 0.04 3.14 ± 0.09 3.93 ± 0.40 84.9 ± 5.3 66.4 ± 3.6 15.6 ± 1.9

(98) Aerodynamic fine particle assessment was done as described in Example 2. Results are shown in Table 10. For all formulation the FPF was calculated to be up to 40% and even up to 60% for the Examples 3B and 3E. In other words, more than 40% of loaded formulations into the device would be deposited in the potential deposition site of inhaled fungal spores after emission from the device. Deposition pattern are exposed in FIG. 7.

(99) Dissolution tests were conducted in the conditions described in Example 1. Every formulations presented different and faster dissolution rate than amorphous spray dried itraconazole (Example 3F) and crystalline bulk ITZ (FIG. 8). As shown in FIG. 9, all dissolution rates of ITZ according to the present invention, 3A to 3E, are at least 5% within 10 minute, 10% within 20 minutes and 40% within 60 minutes when tested in the dissolution apparatus type 2 of the United States Pharmacopoeia at 50 rotation per minute, 37° C. in 9000 milliliters of an aqueous dissolution medium adjusted at pH 1.2 and containing 0.3% of sodium laurylsulfate, namely these dissolution rates are found in the upper area of the curve A which defines the dissolution rate of 5% within 10 minute, 10% within 20 minutes and 40% within 60 minutes. As shown in FIG. 10, the dissolution rates of ITZ according to 3A to 3E are also included in an area between curves B and B′, which defines the dissolution rate of 5% within 5 minute, 10% within 10 minute, 15% within 20 minutes and 40% within 60 minutes, and the one of 50% within 5 minute, 60% within 10 minute, 90% within 20 minutes and 100% within 60 minutes, respectively, when tested in the dissolution apparatus type 2 of the United States Pharmacopoeia at 50 rotation per minute, 37° C. in 9000 milliliters of an aqueous dissolution medium adjusted at pH 1.2 and containing 0.3% of sodium laurylsulfate.

(100) The addition of phospholipids induced an acceleration of the dissolution rate of itraconazole, i.e., >20% of the dissolution ratio at 5 min, >35% at 10 min, >60% at 20 min, >90% at 60 min. Result are shown in Table 11.

(101) TABLE-US-00011 TABLE 11 Dissolution rate of ITZ Dissolution rate of ITZ (%) Formulation 5 min 10 min 20 min 60 min Example 3A (INV) 7.9 11.4 15.2 46.7 Example 3B (INV) 8.1 11.7 16.8 47.3 Example 3C (INV) 6.8 12.7 34.1 98 Example 3D (INV) 24.7 37.2 64.6 96.4 Example 3E (INV) 19.8 36.7 68.3 96.9

(102) Increasing quantity of incorporated phospholipids in the formulation induced acceleration of API's dissolution rate. Indeed, as an example, Example 3C contained 1% (w/w) of phospholipids (expressed by weight of itraconazole) whereas Example 3D contained 10% (w/w). Formulation 3E containing also 10% (w/w) of phospholipids expressed by weigh of itraconazole exhibited a similar dissolution profile than Example 3D, which also contained 10% (w/w) of phospholipids. Although the total amount of phospholipids in the final dry form was much lower for Example 3E (0.99% for Example 3E) this formulation did not show a different dissolution profile than Example 3D which contained a higher total quantity of phospholipids in the final dry form (3.47%).

(103) This indicates that, when evaluated in those conditions, the itraconazole/phospholipids ratio seemed to be the key factor for the API dissolution rate enhancement. It is therefore possible to make vary, to modulate dissolution velocity within this range by varying this ratio. This could be an advantage in vivo to offer different possibility of drug intrapulmonary pharmacokinetic.

(104) Regarding this it is possible to produce a formulation, possessing high fine particle fraction, with a faster dissolution rate than bulk material. But it is also possible to control/modulate this acceleration by varying the quantity of incorporated surfactant.

Example 4

(105) The purpose of this example was to show the ability of the invention to produce matricial dry powders with high fine particle fractions, improved wettability, different dissolution profile and good flow properties using high potentially healthy safe hydrophobic matrix farming agents.

(106) The formulation was prepared at laboratory scale by spray-drying using a Büchi Mini Spray Dryer B-191a (Büchi laboratory-Techniques, Switzerland). A determined quantity of itraconazole, cholesterol and hydrogenated soy-lecithin with more than 90% of hydrogenated phosphatidylcholine (Phospholipon 90H) (see Table 12) were dissolved in 100 ml of isopropanol heated at 70° C. under magnetic stirring (600 rpm). The solution was spray-dried in the following conditions: spraying air flow, 800 l/h heated at 50° C.; drying air flow, 35 m.sup.3/h; solution feed rate, 2.7 g/min; nozzle size, 0.5 mm; Inlet temperature, 70° C.; resulting outlet temperature, 45° C.

(107) TABLE-US-00012 TABLE 12 Composition of the spray-dried solutions in Example 4 Liquid composition Composition (g/100 ml) Example 4 Itraconazole 0.525 g (INV) Cholesterol 1.5 g Phospholipon 90H 0.0525 g

(108) CI value was estimated, as described in Example 1, at 18.9% indicating good powder flowability.

(109) Particle size measurement (Table 13) analysis showed that formulation 4 presented a volume median particle diameter of about 1.1 μm with the Mastersizer2000® and 2.9 μm with the Spraytec®. Some agglomerates seemed to be present in the formulation with higher d(0.9) values. They were probably formed by a certain softening of the phospholipid during the spray drying process due to outlet temperature close of its glass transition.

(110) TABLE-US-00013 TABLE 13 Size distribution parameters measured by laser diffraction methods for the formulation of Example 4 Mastersizer Sirocco 2000 ® Malvern Spraytec ® Formulation d(0.5) d(0.9) d(0.5) d(0.9) N = 3 (μm) (μm) (μm) μm Example 4 1.13 ± 0.03 7.20 ± 1.57 2.94 ± 0.07 9.35 ± 0.19

(111) This presence of agglomerates influenced particles deposition evaluated during aerodynamic assessment of fine particles test realized as described in Example 1. However, 44% of the loaded dose for Example 4 reached the three lower stages of the impactor (table 14).

(112) TABLE-US-00014 TABLE 14 Particle deposition in mg (mean ± SD) and FPF obtained during impaction test (MSLI, 100 l/min, 2.4 sec, 3 discharges per test, nominal dose weighted at 2.5 mg, n = 3). Example 4 Device (mg) 0.73 ± 0.05 Throat (mg) 0.15 ± 0.03 Stage 1 (mg) 0.26 ± 0.14 Stage 2 (mg) 0.17 ± .08  Stage 3 (mg) 0.31 ± 0.03 Stage 4 (mg) 0.50 ± 0.05 Stage 5 (mg) 0.28 ± 0.03 Mean FPD (mg) 1.1 ± 0.1 Mean FPF (%) 44 ± 4 

(113) Dissolution test were performed as described in Example 1 but the dissolution media was constituted of desionized water set at pH 1.2 (HCl 0.063N) containing 1% of sodium lauryl sulfate (FIG. 11). Formulation 4 presented a faster dissolution rate than crystalline micronized bulk itraconazole.

(114) The use of a hydrophobic GRAS matrix former directly modified the release profile of the dispersed API while providing good aerodynamic characteristics and flow properties.

Example 5

(115) The purpose of this example is to show the influence of API's physical state (amorphous Vs crystalline nanoparticles) in the formulation. Two formulations presenting the same quantitative composition were produced and characterized. However the API was in a different physical state in each formulation.

(116) The formulations 5A and 5B were obtained by spray drying a solution or a nanosuspension, respectively, using a Büchi Mini Spray Dryer B-191a (Büchi laboratory-Techniques, Switzerland).

(117) For Example 5A the dry powder was produced by spray drying a feed stock solution of both excipient and API. 0.10 g of itraconazole, 0.9 g of mannitol and 0.01 g of TPGS 1000 were dissolved in 100 ml of an hydro-alcoholic solution (20 water:80 isopropanol) heated at 70° C. under magnetic stirring (600 rpm). This solution was spray-dried in the following conditions: spraying air flow, 800 l/h; drying air flow, 35 m.sup.3/h; solution feed rate, 2.7 g/min; nozzle size, 0.5 mm; Inlet temperature, 90° C.; resulting outlet temperature of 53° C.

(118) For Example 5B the dry powder was produced by spray drying a feed stock solution of excipients in which was re-suspended a determined volume of API nanosuspension added prior spray drying. This procedure was composed of two steps. The first one consisted in size reduction of a micronized API suspension to a nanosize range suspension. The second one consisted to re-suspend a determined quantity of the produced nanoparticles in a feed stock solution containing the matricial agent in order to spray-dry it.

(119) The nanosuspension was prepared as following. In 75 ml of a hydro-alcoholic solution (isopropanol 25:water 50) 75 mg of TPGS 1000 were dissolved under magnetic stirring (600 rp). 750 mg of micronized itraconazole were suspended in this solution using a CAT high speed homogenizer X620 (HSH) (CAT M. Zipperer, Staufen, Germany) at 24,000 rpm during 5 min. The suspension was then circulated in a high pressure homogenizer EmulsiFlex C5 (Avestin Inc., Ottawa, Canada) at 24000 PSI until the particles presented a d(0.5) under 300 nm and a d(0.9) under 2.5 μm. Particle size distribution analysis of the homogenized suspension was done by laser diffraction with a wet sampling system (Mastersizer, Hydro 2000, Malvern instruments, UK). For measurements samples were dispersed in deionized water saturated in itraconazole containing 2% of poloxamer 407 to avoid particle dissolution and aggregation. A refractive index of 1.61 and an absorption index of 0.01 were used for measurements. The high pressure homogenization was done using a heat exchanger, placed ahead of the homogenizing valve to maintain sample temperature below 10° C. 270 ml of a hydro-alcoholic solution composed of 200 ml of isopropanol and 70 ml of water, wherein 2.7 g of mannitol was dissolved under magnetic stirring, was prepared. This solution was kept in an ice bath and 30 ml of the produced nanosuspension was added under magnetic stirring (200 rpm). This final suspension was spray-dried. The following conditions were used during spray-drying: spraying air flow, 800 l/h; drying air flow, 35 m.sup.3/h; solution feed rate, 2.7 g/min; nozzle size, 0.5 mm; Inlet temperature, 80° C.; resulting outlet temperature, 45° C.

(120) The composition of final dry products is shown in Table 15.

(121) TABLE-US-00015 TABLE 15 Quantitative composition of final dry products of Example 5 Quantitative composition Formulation of the dry product Example 5A Itraconazole 9.9% (INV) Mannitol 89.1% TPGS 1000 0.9% Example 5B Itraconazole 9.9% (INV) Mannitol 89.1% TPGS 1000 0.9%

(122) Particle size distribution measurement of the prepared nanosuspension was done. The suspension presented a d(0.5) and a d(0.9) of 0.257+/−0.005 μm and 1.784+/−0.010 μm, respectively. The two dry sample presented good powder flowability. Carr's index values were 19.9% and 24.7% for Examples 5A and 5B, respectively.

(123) PDRX analysis showed that for formulation 5A no characteristics diffraction's peak of crystalline itraconazole were present while the diffractogram of Example 5B exhibited it clearly. Itraconazole was then present in formulation 5A in an amorphous state while it was in a nano-crystalline state in formulation 5B.

(124) Malvern Sirocco® particle size analysis revealed very close size distributions values for both formulations. Results are shown in Table 16. In contrast with those results, Spraytec measurement revealed that after discharge from an inhaler device formulation 5B exhibited a totally different size distribution profile (see in Tables 16). Indeed, the presence of severe agglomerates was observed graphically and traduced by a severe increase of the d(0.9) value to 64.50±19.9 μm.

(125) TABLE-US-00016 TABLE 16 Size distributions parameters measured by laser diffraction with a Malvern Sirocco ® and Spraytec ® for the formulation of Example 5 Mastersizer Sirocco 2000 ® Malvern Spraytec ® Formulation d(0.5) d(0.9) d(0.5) d(0.9) N = 3 (μm) (μm) (μm) (μm) Example 1.60 ± 0.14 3.59 ± 0.25 4.33 ± 0.63 9.12 ± 0.74 5A (INV) Example 1.72 ± 0.07 3.61 ± 0.15 6.30 ± 1.1  64.50 ± 19.9  5B (INV)

(126) Formulation 5B seemed to present lower deagglomeration efficiency than formulation 5A in simulated breath condition. However, despite this presence of severe agglomerates formulation 5B presented the higher fine particle fraction determined as described in Example 1 (see Table 17).

(127) TABLE-US-00017 TABLE 17 Particle deposition in mg (mean ± SD) and fine particle fraction expressed in % of nominal dose (FPF) obtained during impaction test (MSLI, 100 l/min, 2.4 sec, 3 discharges per test, nominal dose weighted at 2.5 mg, n = 3) Example 5A Example 5B Device (mg) 0.27 ± 0.01 0.44 ± 0.02 Throat (mg) 0.49 ± 0.02 0.28 ± 0.01 Stage 1 (mg) 0.24 ± 0.01 0.13 ± 0.03 Stage 2 (mg) 0.37 ± 0.01 0.25 ± 0.04 Stage 3 (mg) 0.62 ± 0.01 0.68 ± 0.03 Stage 4 (mg) 0.31 ± 0.0  0.47 ± 0.02 Stage 5 (mg) 0.04 ± 0.0  0.08 ± 0.0  Mean FPD (mg) 0.95 +/− 0.1 1.19 +/− 0.03 Mean FPF (%) 38 +/− 4  48 +/− 1.2

(128) Dissolution tests were conducted using the method described in Example 1. The two formulations presented different dissolution rates. Formulation 5B exhibited a faster dissolution rate than formulation 5A but the two formulations presented faster dissolution rate than bulk itraconazole.

Example 6

(129) The invention can also consist in a blend of crystalline nanoparticles matricial formulation and the amorphous matricial formulations to vary the dissolution profile of the active ingredient in the desire range. The blend can be realized before or during capsule filling. The burst effect that would be provided by the nanoparticles will induce a determined concentration of ITZ that could be enhanced at a desired velocity by dissolution of the amorphous matricial formulation for which the dissolution rate could be optimized. The proportion of matricial formulation nanoparticle formulation in the final blend will determine to which extend the burst effect (rapid initial dissolution of the drug) would be pronounced.