MXPA00008133A - Liquid crystal forms of cyclosporin - Google Patents

Abstract

This invention relates to novel, liquid crystal forms of the cyclic peptide cyclosporin and to novel powder formulations of cyclosporin prepared using this novel liquid crystal form of the drug. Methods for preparing and using these formulations are also provided. In particular, the present invention relates to dispersible spray dried particles of cyclosporin suitable for pulmonary delivery.

Description

LIQUID CYCLOSPORINE CRYSTALLINE FORMS FIELD OF THE INVENTION This invention relates to a crystalline form of the cyclic peptide cyclosporin and to cyclosporin powder formulations prepared using this novel liquid crystalline form of the drug. Methods for preparing and using these formulations are also provided. In particular, the present invention relates to dry particles of cyclosporin dispersible by spray suitable for pulmonary delivery or administration.

BACKGROUND OF THE INVENTION Cyclosporins are a group of non-polar oligopeptides with immunosuppressive activity. Cyclosporin A, also known as cyclosporine, is the main known cyclosporin, with the structure of cyclosporins B through I also being known (The Merck Index, Twelfth Edition, 464-465 (1996)). Many synthetic analogs of cyclosporin have been prepared. (Id) Cyclosporin A is an orally active immunosuppressant drug that has been used for immune suppression since the mid-1980s (Guzman et al., Pharm Sci. 82: 5) 486-506 (1993)). This has become the support of organ transplant therapy as prophylaxis against organ rejection. The original cyclosporin product for this use, Sandimmune de Sandoz, was formulated in corn oil and designed for oral administration, however, the bioavailability of the gastrointestinal tract tends to be low and also erratic. (Id) Recently, Sandoz has begun marketing an improved patented oral formulation (Neoral) that is claimed to be more reliable than the original (Med. Ad. News, Feb 1996, 7-10). Cyclosporin A produces renal and hepatic toxicity at high doses when administered orally and tolerance should always be verified along with clinical rejection evaluations (Physician's Desk Reference, 52nd Edition, 1891-1901 (1988)). To avoid the complications associated with oral administration or distribution and, in particular, to prevent rejection of lung transplants, it may be desirable to release cyclosporin directly to the lungs. Indeed, nebulized cyclosporin A appears to be effective in the prevention of lung transplant rejection using formulations of liquid ethanol and polyethylene glycol cyclosporin aerosol (Buckart, et al., Inhalation of Delivery of Therapeutic Peptides and Proteins.) Marcel Dekker, NY , pp 281-299 (1997)). Nebulized cyclosporin A also appears to lower oral corticosteroid dependence in asthma (Morley, et al., Cyclosporin Form for Pulmonary Administration, European Patent Application No. 92104426.9 (1992)). The liposomal cyclosporine has also been administered as an aerosol using a nebulizer (Aldrep, et al., Cyclosporin A Liposome Aerosol: Particulate Si ze and Calcula ted Respira tory Deposition, Intl. J. Pharm. 97: 205-212 (1993 )). The main purpose of such formulations has been to decrease the toxicity compared to conventional oral formulations and to provide an alternative to nebulized solutions containing cosolvents. The release or administration of nebulized cyclosporin solution suffers from limited solubility of the drug in vehicles based on aqueous solvents. In addition, there are safety concerns about the nebulization of organic vehicles. The release or administration of nebulized solutions and suspensions suffers from a low drug release efficiency of commercial nebulizers. The aerosolization of cyclosporin A (CsA) with MDI would involve a CsA solution in propellants (chlorofluorocarbon propellants or without chlorofluorocarbon) or the use of finely divided CsA suspended in propellants. The poor drug release efficiency and low drug loading capacity makes MDIs inconvenient means of aerosol release for human dosage regimens that may require 1 mg to 20 mg of CsA released per day to the lungs. In view of the difficulty of distributing a cyclosporine solution by inhalation, it may be desirable to distribute the cyclosporin as a dry powder. The ability to distribute pharmaceutical compositions as dry powders, however, is problematic in certain aspects. The dosage of many pharmaceutical compositions is often critical, so that it is desirable that dry powder distribution systems be able to accurately, accurately and reliably deliver the intended amount of drug. It is also essential that dry powders for lung distribution be readily dispersible to ensure adequate systemic distribution and absorption. Because CsA can cause gingivitis, it is important that oropharyngeal deposition be minimized.

BRIEF DESCRIPTION OF THE INVENTION The present invention provides a novel liquid crystalline form of the cyclosporin previously not known. This novel form is thermotrophic liquid crystalline cyclosporine. It has been unexpectedly found that spray-dried organic solutions containing cyclosporin, in particular cyclosporin A (CsA), under specific conditions, result in this novel form of cyclosporin. Spray drying of organic solutions containing cyclosporin produces powders where the particular cyclosporin exhibits an absence of three-dimensional order (3-d) as determined by powder X-ray defamation (PXRD) and also exhibits an order 2- d when analyzed by small-angle X-ray scanning (SAXS). In addition, it exhibits a phase change from solid to liquid over a narrow temperature range with a gradual change in heat capacity, i.e., a fusion similar to the glass transition. This form of cyclosporine is liquid cyclosporine. The conditions of the spray drying process can be varied within certain limits to vary very narrow particle size distributions which make the powders especially suitable for efficient distribution by oral inhalation. These powders have a high distribution efficiency when aerosolized with a dry powder inhaler and have demonstrated physical, chemical and aerosol stability during prolonged periods of high temperature and high humidity. In one aspect, the invention provides liquid crystalline cyclosporine. In particular, the liquid crystalline form of cyclosporin A is provided. In another aspect, the invention provides dispersible powder formulations of liquid crystalline cyclosporine for pulmonary distribution. In particular, dispersible powder formulations based on cyclosporin are provided, which are spray-dried from cyclosporin and, optionally, excipient, in a solvent, as well as the methods for producing those formulations. Specifically sprayed cyclosporin A powders are specifically provided. In a further aspect, the invention provides methods for treating a subject suffering from or being the subject of a condition which can be alleviated or prevented by administration of cyclosporin, which comprises administering the dispersible powder cyclosporin formulations described above. In particular, methods are provided to alleviate or prevent lung diseases or conditions that affect the lungs. Cyclosporine can be used as an anti-inflammatory, immunosuppressant or anti-asthmatic agent.

BRIEF DESCRIPTION OF THE DRAWINGS FIGS .A through IC illustrate the difference in powder X-ray diffraction patterns between two of the known crystalline forms of cyclosporin (non-solvated orthorhombic (Figure IA) and tetragonal dihydrate (Figure IB)) and the novel thermotropic liquid crystalline form of the cyclosporin provided by the present invention (Figure IC). Figure 2A illustrates a representative open tray differential scanning calorimetry (DSC) trace and Figure 2B illustrates a closed tray DSC trace representative for the thermotrophic liquid crystalline form of the cyclosporin of the present invention. Figure 3 presents the small-angle X-ray diffraction data for orthorhombic and tetragonal crystalline cyclosporine and for liquid crystalline cyclosporine spray dried at 10, 80 and 150 ° C. Figure 4 shows the dielectric analysis (DEA) of the thermotrophic liquid crystalline CsA formulation according to the present invention.

Figures 5A, 5B and 5C illustrate analysis by Cyclosporin A CLAP spray-dried under accelerated storage conditions of 110 ° C for 196 hours, 140 ° C for 50 hours and 210 ° C for 10 minutes, respectively. Figures 6A, 6B and 6C show that cyclosporin A powder is spray-dried for 10 months at 40 ° C and a relative humidity of 75% showed no appreciable degradation based on CLAP analysis. Figure 7 demonstrates that spray dried cyclosporin A powders stored at room temperature for 15 months showed no appreciable degradation based on CLAP analysis.

DETAILED DESCRIPTION OF THE INVENTION The present invention is based at least in part on the discovery of a novel thermotropic liquid crystalline form of cyclosporin, particularly cyclosporin A. This liquid crystalline form of cyclosporin can be formulated as a dispersible powder. spray drying of organic solvents. Cyclosporin-based compositions are suitable for pulmonary distribution due to their dispersibility characteristics. The compositions of the invention are easily aerosolized and presented to the deep lung of a host when they are distributed by means of a dry powder inhaler. The powder formulations of the present invention retain their stability, are easily dispersible for pulmonary distribution and allow the packing of unit doses. The invention consists, in part, of compositions comprising cyclosporin in dispersible powder formulations. The use of particles of a certain size range allows the distribution of cyclosporine to the alveolar area of the lungs (ie, to the deep lung). Optionally, the powder formulations of the present formulation may contain stabilizers and excipients such as buffer salts, sugars, toning agents, preservatives and antioxidants. The compositions of the present invention are useful in dry, pulmonary, powder drug delivery systems, including but not limited to those described in US Patent No. 5,458,135 and International Patent Publication O96 / 09085. The solid state forms of CsA that have been previously reported are described in Table 1.

Table 1. Solid State Forms of Cyclosporin A Solvation System Group Interval Unit Cell Dimensions Reference Crystalline Fusion (° C) Spatial (Á) tetragonal dihydrate 140-150 P4j 13,837 12,387 41,242 1.2 tetragonal unsolvated P4j nr nr ortho-bico di- (di- -150 P2l2121 12.5 22.9 23.4 isopropyl ether) orthorhombic no 180-1195 P2J.2J.2I 12.7 15.7 36.3 1.3 solvated orthorhombic monohydrate nr P2J.212J 12.5 22.9 28.4 4.3 amorphous n / a n / a n / a n / a n / a n / a nr = not reported n / a = not applicable Reference rón, et al., Orthorhombic Ci closporin citud of Patent of the United Kingdom No.

Osli, et al., The Conformation of the Crystals and in Soluction, Helvetica 682-704 (1985). tha, et al., The Story of the Czech, 28 pp. 10 Knott, et al., Neutron Structure of the San t Cyclosporin A, Acta Crys., C46: 1528- crystalline and CsA crystalline forms exhibit high melting points and characteristic sharp diffraction when by PXRD and are birefringent to polarized light. Amorphous materials, unlike liquid crystals, have no peaks by SAXS and are not We have discovered that CsA powders prepared by spray drying organic solvents can be designed so that they are not of the orthorhombic or tetragonal crystalline forms. In fact, sharp diffraction peaks indicative of a three-dimensional order by PXRD of the spray-dried powders were not observed (Figure IC). However, spray-dried powders do not exhibit a two-dimensional order when analyzed by SAXS, indicative of a liquid crystalline form (Figure 3). In addition, CsA powders prepared by spray drying of organic solvents also showed a different endotherm between 0-75 ° C when analyzed by differential scanning calorimetry (DSC) at 10 ° / min in an open tray (Figure 2A). Since the physical nature of the powders does not change (not a melt) through the temperature range and because the endotherm is absent in the closed tray DSC trace (Figure 2B), it is believed that the transition should be to the evaporation of the solvent. A fusion occurs starting at approximately 120 ° C when analyzed by hot stage microscopy at 2"/ minute.The DSC thermogram at the same heating rate requires a gradual change in heat capacity (Cp) (a transition similar to Tg) at that temperature., crystals ordered in 3-d do not exhibit a gradual change of Cp going from solid to liquid. In addition, the dielectric analysis is consistent with the results of the DSC and confirms that the observed transition at ~ 120 ° C is a second order transition, as demonstrated by the dependence of the observed frequency.

A. Definitions As used herein the following terms have the following meanings: The terms "dispersibility" or "dispersible" mean a dry powder having a moisture content and / or residual solvent of less than about 10% by weight (% p ), usually less than about 5% p and preferably less than about 3% p and often less than about 1% p; a particle size between 0. 1 and 15 μm, often between 0.2 μm and 10 μm, usually an average basic diameter (MMD), of about 0.4 to 5 μm, preferably a MMD of about 1 to 4 μm and most preferably a MMD of 1 at 2 μm; a released dose of more than about 30%, usually greater than about 40%, preferably greater than about 50% and more preferably greater than about 60%; and a particle size distribution with an average mass aerodynamic diameter (MMAD) of about 1-5 μm, usually an MMAD of about 1.5-4.5 μm and preferably an MMAD of about 1.5-4.0 μm, or with at least about 40% (preferably at least about 50%) of the particles with a diameter of less than about 3.3 μm. The term "cyclosporin" means any of the group of non-polar cyclic oligopeptides with immunosuppressive activity and includes known cyclosporins A through I. In particular, this term includes cyclosporin A, also known as cyclosporin. Synthetically produced, derived or naturally purified and recombinantly produced entities are included, as are the analogs, derivatives, agonists, antagonists and pharmaceutically salts of any of these. The term also includes cyclosporins which have D-amino acids, modified amino acids, derivatives or unnatural in the D or L configuration and / or peptomimetic units or prodrugs as part of their structure. A thermotrophic liquid crystal is a state of matter different from the amorphous and three-dimensional crystalline states and is characterized by the existence of a long range order in one (nematic) or two (smectic) dimensions in the absence of solvent. An amorphous phase lacks a long range order and a three-dimensional crystal phase contains a large three-dimensional order. The term "powder" means a composition consisting of finely dispersed solid particles that are freely flowing and are capable of being easily dispersed in an inhalation device and subsequently inhaled by a subject, so that the particles reach the lung spaces deep to allow deposition in the alveoli. In this way, it is said that the dust is "breathable". The terms "pharmaceutical excipient" or "additive" means compounds which solubilize cyclosporin and / or improve the performance and stability of the powder aerosol. The types of excipients useful in the present invention include buffer salts, sugars, toning agents, preservatives and antioxidants and the like. The term "physically stable" or "physical stability" is intended to refer to a composition that does not show a phase change with time. The term "chemically stable" or "chemical stability" means that the composition shows less than 10% and, preferably, less than 5% total degradation in 2 years at room temperature at the specified storage conditions. "Aerosol stability" means that the aerosol composition shows no statistical change in the efficiency of the dose distributed over time. The term "subject" includes any human or animal species that needs cyclosporine for the treatment or prophylaxis of conditions for which the pulmonary distribution of cyclosporin would be effective.

B. Compositions: The present invention is directed to liquid crystalline forms in cyclosporin and to powder compositions containing cyclosporin, dispersible, suitable for pulmonary distribution formed by spray drying of organic solvents. The dispersible powder compositions comprise a therapeutically effective amount of cyclosporin, optionally in combination with a pharmaceutically acceptable excipient carrier.

Spray drying is a process that uses high temperatures and high rates of carrier gas flow to rapidly evaporate solvents from a solution containing dissolved solutes. The solvent evaporates to leave a solid particle. The size of the particle depends on the conditions of the spray drying processes (eg solids content of the solution, atomization gas pressure, atomizer nozzle design and cyclone collector design). The control of particle size and particle size distribution is important for efficient inhalation distribution to the airways of the deep lung. The average mass diameter (MMD) of the particles should preferably be between 1 and 2 μm, with 100% of the particles being less than 15 μm.

It can also be difficult to control the particle size and the particle size distribution in the compositions produced by spray drying. For the lung distribution it is critical that the average particle size is maintained in a respirable range so that the amount of particles comprising the composition outside the target size range is minimized. In addition, it may sometimes be difficult to achieve a desirable low residual solvent and / or moisture content for physical and chemical stability in the final particulate product, particularly economically. Useful methods are described, for example, International Patent Application No. l PCT / US97 / 07779, the description of which is hereby incorporated by reference in its entirety. Cyclosporin, including cyclosporin a (CsA) is highly hydrophobic and is practically insoluble (< 6 μm / mL) in aqueous vehicles. Organic solvents with boiling points of less than 200 ° C, preferably less than 150 ° C, more preferably less than 100 ° C, should be used to obtain powders with low residual solvent. Preferred solvents for producing cyclosporin (including CsA) spray-dried powders include, but are not limited to, ethanol, acetone, acetonitrile, methanol, isopropanol, and methylene chloride, alone or in combination or in cosolvent systems. Pharmaceutically acceptable protic solvents with low dielectric constants are most preferred (eg, ethanol is preferred over methanol). Mixtures of solvents with less than 50%, more preferably less than 25%, more preferably with 10% less water by volume can also be used to spray off cyclosporin. The use of water in a mixture of solvents allows the incorporation of water-soluble excipients in the CsA particles, however non-aqueous systems are preferred. Water-soluble excipients useful in the present invention include, but are not limited to, buffer salts (e.g., citric acid-sodium citrate), natural and synthetic sugars as diluting agents (e.g., lactose, mannitol), toning agents ( for example, sodium chloride) and preservatives and antioxidants (for example, ascorbic acid / sodium ascorbate). The excipients and / or pharmaceutical additives generally used in the present invention include suitable pH adjusters or buffers, such as organic salts prepared from organic acids and bases, such as sodium citrate, glycine, sodium tartrate, sodium lactate. , tromethamine and the like. Proteins (eg, HSA, recombinant human albumin (rHA), gelatin and casein), peptides (eg, aspartame) and amino acids (eg, alanine, glycine, arginine, glutamic acid and aspartic acid), which improve the dispersibility of the powder may be useful. Also useful are carbohydrates / sugars and alditols. Suitable carbohydrate / sugar compounds include sucrose, trehalose, lactose, raffinose, and the like. Suitable alditols include mannitol and pyranosyl sorbitol and the like. Polymeric excipients / additives include polyvinylpyrrolidones (PVP), Ficolles, soluble hydroxy ethyl starch, dextrata and the like of high molecular weight. Also useful are small amounts of pharmaceutically acceptable surfactant, such as Tweens, chelants such as EDTA, and inorganic acids and bases such as sodium phosphate and the like. Other excipients and / or pharmaceutically suitable additives include those described in Remington, Pharmaceutical Sciences 18th edition, (1990), the disclosure of which is incorporated herein by reference. The temperatures used to dry the drops of spray solution can range from 20 to 300 ° C, preferably from 30 to 150 °, and more preferably from 40 to 120 ° C. These temperatures are expressed as at the outlet temperature of the carrier gas. Specifically, the outlet temperature is the temperature of the gas at the outlet of the drying chamber before entering the cyclone and the collector. Correspondingly, higher temperatures are required at the spray point to reach the recommended temperatures. The spray-drying process may include keeping the powder for an additional period of time at a given temperature after completing the feeding of the cyclosporin solution through the system (i.e., secondary drying). This secondary drying can be used to reduce any residual solvent left in the powder. A drop size with a diameter of about 4 to about 8 μm is preferred to achieve optimum powder characteristics. Such droplet size can be achieved, for example, by using the atomization method described in International Patent Application No. PCT / US97 / 07779. Unless otherwise specified, atomization methods that result in droplet sizes of 4-8 μm were used in the following examples. The average mass diameter (MMD) of the powders prepared by organic spray drying was measured by centrifugal sedimentation with a Horiba CAPA-700 Particle Size Analyzer. A sample of dust was dispersed in a Sedisperse -ll vehicle (Micromeritics, Norcross, GA), which was presaturated with cyclosporin A and filtered before the addition of the powder sample. The particle size ranged from a MMD of about 0.7 to about 2.4. The size of the CsA particles was confirmed by scanning electron microscopy; it was also found that the particles were generally spherical in shape, that is, from smooth to dimpled, raised or wrinkled spheres.

The aerosol performance characteristics of the powders were evaluated using the aerosol device of the therapeutic inhalation system. The device includes an aerosol chamber and employs a volume of compressed air to disperse the powder from a pack of aluminum foil ampoules. The efficiency of the distributed dose (DDE) for each powder was defined as the percentage of the nominal dose contained within the packet of ampoules that exited the nozzle of the aerosol device and was captured on a filter through which vacuum was extracted ( 30 L / min) for 2.5 seconds after operating the device. The filter was weighed before and after the drive of the device to determine the mass of powder distributed through the nozzle. The particle size distribution of the aerosolized powders was determined using an Andersen cascade impact device through which a vacuum was extracted (28.3 L / min) for 2.5 seconds.

The solvent and / or residual content of the powder particles of the present invention is usually less than about 10% by weight, preferably less than about 5% p and more preferably less than about 3% p. Powders with such a low solvent and / or moisture content are generally physically and chemically stable during storage at room temperature and are easily dispersible in an inhalation device to form an aerosol. Stability studies were performed on the spray-dried cyclosporin formulations of the present invention, and showed that these compositions retained aerosol stability and physical stability. In particular, the DDE of a cyclosporin powder dried by ethanol spray at 70 ° C without secondary drying was measured immediately after the preparation and found to be 48.4%. The powder was then stored at room temperature for 10 months. Again the DDE was measured and found to be 49.5%, indicating that the powder retained the stability of the aerosol. In another stability test, the DDE of the cyclosporin powders dried by ethanol spray at 70 ° C without secondary drying was measured immediately after the preparation and found to be 72%. The powder was then stored at the accelerated conditions of 40 ° C and 75% relative humidity (RH). The DDE was measured 8 weeks later and again 15 weeks after storage under these conditions. The results showed that the DDE remained approximately the same, that is, approximately 75% at 8 weeks and approximately 74% at 15 weeks. A DSC scan of this powdered formulation made immediately after the preparation showed a melt at 118.61 ° C, while a DSC scan of the formulation made 15 weeks later at 40 ° C and 75% relative humidity showed a 119 ° C. These results indicate that no physical changes of the powder occurred during this period and that the powder retained the stability of the aerosol. The amount of cyclosporin that constitutes a therapeutically effective amount will vary in composition depending on the biological activity of the cyclosporin employed and the amount needed in the unit dosage form. The condition to be treated or prevented will also determine the amount of cyclosporine required, as well as the subject to which the composition is administered. The compositions comprise at least about 40% by weight of cyclosporin in the formulation, preferably between about 70% to about 100% and more preferably from about 90% to about 100%. The amount of pharmaceutically acceptable excipients and additives may be from about 0-60%, preferably from about 0-30% and most preferably from 0-10% by weight.

The compositions of the present invention will often be packaged as unit doses where a therapeutically effective amount of the cyclosporin composition is present in a unit dose receptacle, such as a pack of ampoules, gelatin capsule, or the like, as long as the Provide a barrier against moisture. The powder, dry compositions of the cyclosporin base of the present invention can be produced by spray-drying solutions or suspensions of the cyclosporin, and, optionally, excipients, in a non-aqueous solvent under conditions to provide a respirable dry powder. Solvents may include ethanol, acetone, acetonitrile, methanol and isopropanol, which can be easily dried. In addition, dry, cyclosporin-based powder compositions can also be produced by evaporative drying, lyophilization, cooling of a melt, precipitation, including precipitation of supercritical fluid.

C. Characterization: It was found that, by spraying cyclosporin from organic solvents, a thermotropic liquid crystal of cyclosporin was formed. In particular, the characterization of this form of cyclosporine using polarized light microscopy showed that it was refringent, indicating that it was a non-amorphous form of cyclosporine. Similarly, the SAXS analysis showed the presence of sharp peaks (Figure 3), a characteristic of non-amorphous materials.

Further characterization of this novel form of cyclosporin by powder X-ray diffraction revealed non-acute diffraction peaks, which would have indicated a three-dimensional order, such as that found in crystalline structures, indicating that this was not a crystalline form three-dimensional cyclosporin. Figures A through I C show X-ray diffraction patterns of the powder of the crystalline forms of cyclosporin (tetragonal and orthorhombic) and our novel spray-dried form. Hot stage microscopy, DSC, DEA and SAXS were used to characterize the novel form of cyclosporin. The hot stage microscopy of this material showed a different melting point, which is characteristic of both crystalline and liquid crystalline materials. The DSC showed a gradual change in the heat capacity at the melting temperature, characteristic of liquid crystalline materials. Figure 2A shows a DSC (heating at 10 ° C / min) of the liquid cyclosporin of the present invention, which indicates a melting point that starts at 122 ° C and may be between 115 ° C and 125 ° C. When analyzed by DEA, it was found that such a transition depended on frequency, suggesting that this was actually a second-order transition and not a true merger. The crystalline liquid state was confirmed by SASX, which showed sharp diffraction peaks at low diffraction angles, as is characteristic of the 2 dimensional order in liquid crystals. Figure 2 shows the SAXS for spray-dried cyclosporin. The material remains a liquid crystal below the melting (10 ° C and 80 ° C) and above the melting (at 150 ° C).

D. Pulmonary Ciclosporin Pulmonary ciclosporin is useful for the treatment of asthma and lung transplants but has the potential to be used in many other indications as well. Pulmonary cyclosporine may be useful to treat sarcoidosis. Obliterative bronchiolitis (OB), the lung pathology that occurs in the rejection of lung transplantation, also occurs in the rejection of heart and bone marrow, but there is the potential that inhaled cyclosporine is of use in other transplant therapies in conjunction with oral immunosuppressants. Chronic inflammatory disease of the lung, chronic obstructive pulmonary disease, emphysema, primary and secondary pulmonary hypertension, cystic fibrosis, pulmonary infections or idiopathic pulmonary fibrosis (IPF) are after lung diseases that can respond to inhaled cyclosporine, since they seem to be caused or exacerbated by an overactive immune system. In addition, cyclosporine may be useful for treating pulmonary complications associated with autoimmune diseases such as rheumatoid arthritis. The advantage of circulating pulmonary cyclosporin for diseases or conditions that affect the lungs is that the total body amount of drug can be reduced, which reduces or eliminates systemic side effects. According to the present invention, cyclosporin can be distributed directly to the deep lung in the form of a dry pr using a dry pr delivery device. A significant requirement for such dry pr distribution devices is efficiency. The dose distributed should be relatively high to reduce the number of inhalations required to achieve a total dose. The ability to achieve adequate dispersion is a significant technical challenge that requires in part that each unit dose of pr consumption be dispersed easily and reliably. Certain pulmonary distribution devices, such as those described in U.S. Patent No. 5,458,135 and International Patent Publication O96 / 09085 (the descriptions of which are incorporated herein by reference), are useful for the pulmonary distribution of dry pr drugs. . The description of each of the publications, patents or patent applications mentioned herein is therefore incorporated by reference in its entirety to the same extent as if the language of each publication, patent and individual patent application was specifically and individually incorporated. here as a reference DESCRIPTION OF EXAMPLES OF THE INVENTION The following examples are not intended to limit the scope of the invention in any way.

Materials and Methods: In general, the following materials and methods were used in the following examples, unless otherwise indicated.

Materials: Cyclosporin A, grade GMP, was obtained as a crystallized acetone powder (melting point 148-150 ° C) from Poli Industria Chimica, S.p.A.

Storage of the Sample: The spray dried powders were stored under a dry atmosphere (RH <5%).

Physical Methods: Powder particle size distribution The particle size distribution (PSD) of the spray-dried powder samples was measured with a centrifugal sedimentation particle analyzer CAPA-700 from Horiba. A powder sample was dispersed in a Sedisperse W-ll vehicle (Micromeritics, Norcross, GA), which was presaturated with cyclosporin A and filtered prior to the addition of the powder sample. Approximately 5 mg of powder was suspended in approximately 5 ml of the Sedisperse and sonicated for 5-10 minutes in a Lab Supplies ultrasonicator before analysis. The instrument was configured to measure a particle size range of 0.4 to 10 μm in diameter using a particle density of approximately 1.2 g / cm3.

Powder X-ray diffraction (PXRD) The PXRD was performed on a D-500 X-ray Diffractor from Siemens. The sample was measured at 3 ° / min (0.8 segmentation / 0 .04 ° step) and 0.5 ° / step, 1 sec / step, respectively. The scan was carried out continuously from 2 ° to 2 °.

Small-angle X-ray diffraction (SAXS) The SAXS analysis was performed on a 12KW diffractometer from Rigaku equipped with a Kratky camera and a 20 cm position-sensitive detector from Braun, using a copper X-ray source of 1542 Á at a scanning speed of 0.12 ° / min in the range of 0 to 2.2 ° in 2T.

Method for testing birefringence Birefringence was tested using a polarized light microscope, Nikon Optihot 2-Pol, equipped with a Hamamatsu camera and a C2400 controller. The images were printed on a UP890MD graphic printer. The powders, either dried or immersed in Sedisperse W-ll or water, were examined under the 20A, 40X and 60X targets, and photomicrographs were taken through a normal and cross-polarized light.

Method for hot stage microscopy Hot stage microscopy was performed on a Nikon Optiphot 2-Pol equipped with a Metler Toledo FP82HT Hot Plate and a Humamatsu camera and a C2400 controller. Photomicrographs were printed on a UP890MD graphic printer. The slide was placed on the hot stage and a representative field was found using the 40X objective. The samples were heated at a rate of 2 ° C / min from room temperature to above the melting point. The images were taken when the changes were observed visually.

TGA Residual Solvent Method The samples were analyzed by Oneida Research Services, Inc. The TGA was run on an Omnitherm 1500. The samples were heated at 30-200 ° C / min under a nitrogen atmosphere with a flow rate of 30. ml / min. The weight loss due to drying was measured and presented as the loss of% by weight.

TGA Decomposition Method Samples were analyzed by Oneida Research Services, Inc. The TGA was performed on an Omnitherm 1500 TGA. The samples were heated from 40-600 ° C / min at 10 ° / min under a nitrogen atmosphere with a Flow rate of 30 ml / min.

Differential Scanning Calorimetry (DSC) The scans were performed on a Modulated TA Instrument model 2920 DSC with a refrigerated cooling system unit (RCS) TA, and pure helium gas with a flow rate of ~ 120 cm3 / min. The cell flow rate was set at approximately 40 cm 3 / min. The scans were performed at 10 ° C / min unmodulated, with an equilibrium temperature of -30 ° C for 15-30 min, followed by heating at 200-225 ° C. Open and closed aluminum trays were filled with approximately 2 mg-6 mg of powder (Figures 2A and 2B, respectively).

Dielectric analysis (DEA) The DEA scan was performed on a TA Instrument 2970 Dielectric Analyzer using hydrogen to cool the sample to the initial temperature. The powder (40-45 mg) was pressed into a tablet of 1/2 inch (1.27 cm) in diameter and a thickness of approximately 0.3 mm, using a Carver press for 40 seconds at 1 ton. Two thin layers of Teflon of 25 μm and 7/16 inch (1.11 cm) in diameter were inserted into the matrix of the tablet to alleviate adhesion to the faces of the matrix. During the measurement, the pellet was surrounded by a silicone gasket, ID 9/16 inches (1.42 cm), OD 1 1/16 (2.69 cm) thick, 0.35 mm, to help maintain the thickness during the measurement. The tablet was also sandwiched between two layers of Teflon, 25 μm thick, to remove possible contributions of ionic conductivity. The electrons were fabricated from TA Instrument sensors for samples covered by 25 μm thick sputter and gold sheet particle sputtering, a 7/16 inch (1.11 cm) diameter disc and the other 6/8 inches (1.90 cm) in diameter, so that the sample only came into contact with the leaf-shaped electrodes. The experiments were carried out using a heating rate of 2 ° C / min from -40 to 200 ° C and using the following frequencies: 1 Hz, 10 Hz, 100 Hz, 1,000 Hz, 10,000 Hz, 100,000 Hz. The permisivities measured with this method was in arbitrary units since the data was not adjusted for the gold plate sensors; however, this has no impact on the interpretation of the results (Figure 4).

Aerosol Methods: Distributed dose test The distributed dose test was performed to determine the efficiency and reproducibility of the pulmonary distribution of the dispersible powder cyclosporin compositions. The performance characteristics of the aerosol were evaluated using the aerosol device of the inhalation therapeutic system, similar to the devices described in WO96 / 09085. The device includes an aerosol chamber and employs a volume of compressed air to disperse the powder from a pack of aluminum foil blisters. The efficiency of the distributed dose (DDE) for each powder was defined as the percentage of nominal dose contained within a packet of ampoules that exhibited the nozzle of the aerosol device and was captured on a glass fiber filter (Gelman, 47). mm diameter) through which a vacuum was drawn (30 L / min) for 2.5 seconds after the device was operated. The efficiency of the distributed dose was calculated by dividing the mass of the dust collected on the filter by the mass of the powder in a packet of powder.

Each result was the average of 5-10 repeated measurements.

Aerosol particle size distribution The particle size distribution of the aerosol obtained using an eight-stage cascade impact device (Graseby Andersen, Smyrna, Georgia). The airflow of the impact device was set to extract a vacuum of 28.3 L / min, the flow rate calibrated for the instrument, for 2.5 seconds. For each test, packages of ampoules filled with approximately 5 mg of powder from the inhaler were dispersed. The particle size was determined by weighing the powder on the plates of the impact device and evaluating the results on a logarithmic probability plot. Both the average basic aerodynamic diameter (MMAD) and the basic fraction of less than 5 μm were determined from the logarithmic probability plot.

Chemical Stability Method The CLAP method that indicates the stability that was used is described in Oliyai, et al., Kinetics of Acid-Ca talized Degradation of Cyclosporin A and its Analogs in Aqueous Solutions, Peptide and Protein Res. 43: 239 -247 (1994). The method was carried out as described, however, the ratio of the mobile phase was adjusted slightly to obtain the retention times for cyclosporin A and iso-cyclosporin A.

Example 1: Cyclosporin A Spray-dried Ethanol at 70 ° C Without Secondary Drying Solution Preparation 1.5 g of cyclosporin A were dissolved in 50 mL of ethanol (200 test, USP, NF grade Spectrum).

Spray drying A dry powder comprised of cyclosporin A or spray-dried organic solution was produced using a Buchi B-190 laboratory spray dryer using a nitrogen atmosphere containing less than 5% oxygen (with N2 atm < 5% 02) with the following parameters: Exit Temperature 70 ° C Inlet Temperature 100 ° C Feed Speed 5 mL / min Atomizer Flow Rate 13 lit / min Secondary Drying None Powder Characterization The PXRD of the powder showed that it exhibits a double halo with two maximums at 2? equal to 8.5 ° and 17 ° (Figure IC). The absence of sharp peaks in the diffractogram shows that the material is not a three-dimensional crystal. Polarized light microscopy showed that the particles are birefringent. The SEM images look like rounded and highly rough particles. There was no change of solid state to the formulation due to the increase in the relative unit according to that shown by the DSC of the powders exposed to a relative humidity of 0% and a relative humidity of 75%. The DSC scan showed a large endotherm ranging from about 20 ° C to about 70 ° C with a maximum peak at 69 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 122 ° C. The hot stage microscopy showed that the fusion is in the range of 138-140 ° C. It was determined that the MMD of the powder sample was 1.6 μm, with 96.5% less than 5.2 μm. The AED showed a slope change of the frequency in the permissiveness indicative of a second-order transition, in the same temperature range as determined by the DSC, where the change in heat capacity began around 125 ° C. At the temperature interval at which the largest endotherm was observed in the DSC exploration (~ 20-70 ° C), there was no change in the permissiveness, suggesting that the endotherm is not due to a phase change but to the evaporation of solvent.

Aerosol Characterization It was determined that the efficiency of the distributed dose (DDE) of cyclosporin A powder dried by previous spray was 79% ± 4.2% (n = 10). It was determined that the average basic aerodynamic diameter (MMAD) was 2.81 μm with 85% less than 5 μm.

Chemical Stability The analysis by CLAP showed degradation products of cyclosporin A spray-dried under stress conditions. Figure 5A is a sample stored at 110 ° C for 196 hours. Figure 5B shows a sample stored at 140 ° C for 50 hours and Figure 5C shows a sample stored at 210 ° C for 10 minutes. The degradation products of liquid crystalline cyclosporine are different from those of other reported forms of cyclosporin (Oliyai, et al., Kinetics of Acid-Ca talized Degradation of Cyclosporin A and its Analogs in Aqueous Solutions, Peptide and Protein Res. 43: 239-247 (1994) and Oliyai, et al., Kineti cs and Mechanism of Isometry of Cyclosporin A. Pharm, Res. 9 (5): 617-622 (1992)).

Example 2: Cyclosporin A Acetone Dewdrop At 88 ° C With Secondary Drying Preparation of the Solution 1.5 g of cyclosporin A were dissolved in 50 mL of acetone (CLAP grade of J.T. Baker).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Output Temperature 88 ° C Input Temperature 118 ° C Feed Speed 5 mL / min Atomizer Flow Rate 13 lit / min Secondary Drying 85 ° C / 5 min.

Once the solution was consumed, the outlet temperature was maintained at 85 ° C for 5 minutes by slowly lowering the inlet temperature to provide secondary drying.

Powder Characterization The PXRD of the powder showed that it exhibited a double halo with two maxims 2? equal to 8.5 ° and 17 °. Polarized light microscopy showed that the particles were birefringent. The TGA analysis of the powder showed that the powder had 0.1% by weight of residual solvent and a decomposition temperature range of 347-421 ° C. The SEM image of the powder showed that the particles were rounded, with slight dimples. The DSC scan showed a large isotherm ranging from about 20 ° C to about 70 ° C with a maximum peak at 58 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at the initial temperature of 121 ° C. It was determined that the MMD of the powder sample was 1.19 μm, with 95.8% less than 5.2 μm.

Characterization of the Aerosol The DDE of the cyclosporin A powder dried by previous spray was determined to be 59% ± 9% (n = 10). It was determined that the MMAD was 2.0 with 84% less than 5 μm.

Example 3: Cyclosporin A Dried by Ethanol At 85 ° C With Secondary Drying Preparation of the Solution 1.5 g of cyclosporin A were dissolved in 50 mL of ethanol (200 test, USP, Spectrum NF grade).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Exit Temperature 85 ° C Inlet Temperature 120 ° C Feed Speed 5 mL / min Atomizer Flow Rate 13 lit / min Secondary Drying 85 ° C / 5 min.

Once the solution was consumed, the outlet temperature was maintained at 85 ° C for 5 minutes by slowly lowering the inlet temperature to provide secondary drying.

Powder Characterization The PXRD of the powder showed that it exhibited a double halo with two maxims 2? equal to 8.5 ° and 17 °. Polarized light microscopy showed that the particles were birefringent. TGA analysis of the powder showed that the powder had 0.3% by weight of residual solvent and a decomposition temperature range of 348-425 ° C. The SEM image of the powder showed that the particles were raisin-like. The DSC scan showed a large isotherm that fluctuated from about 20 ° C to about 70 ° C with a maximum peak at 62 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 122 ° C. It was determined that the MMD of the powder sample was 1.27 μm, with 100% less than 5.2 μm.

Characterization of the Aerosol It was determined that the DDE of the cyclosporin A powder dried by previous spray was 71.4% ± 6% (n = 10). It was determined that the MMAD was 2.8 with 86% less than 5 μm.

Chemical Stability The CLAP analysis showed no appreciable degradation of samples of spray dried cyclosporin A powder stored at 40 ° C and a relative humidity of 75% for 10 months (see Figures 6A, 6B and 6C). The powder was considered to be chemically stable for the duration of the study. Figure 6A is a chromatogram of the mobile phase. Figure 6B is a chromatogram of cyclosporin A in the mobile phase with a loading of 25 μg. Figure 6C is a chromatogram of the spray-dried cyclosporin A aged at 40 ° C and a relative humidity of 75% for 10 months reconstituted in the mobile phase.

Example 4: Cyclosporin A Acetonitrile Dewdrop At 101 ° C With Secondary Drying Preparation of the Solution 1.5 g of cyclosporin A were dissolved in 50 mL of acetonitrile (CLAP grade of Burdick and Jackson).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Output Temperature 101 ° C Input Temperature 141 ° C Feed Speed 5 mL / min Atomizer Flow Rate 13 lit / min Secondary Drying 100 ° C / 5 min.

Once the solution was consumed, the outlet temperature was maintained at 100 ° C for 5 minutes by slowly lowering the inlet temperature to provide secondary drying.

Powder Characterization The PXRD of the powder showed that it exhibited a double halo with two maxims 2? equal to 8.5 ° and 17 °. Polarized light microscopy showed that the particles were birefringent. The SEM image of the powder showed that the particles had slight dimples. The DSC scan showed a large isotherm that fluctuated from about 20 ° C to about 70 ° C with a maximum peak at 69 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 121 ° C. It was determined that the MMD of the powder sample was 1.99 μm, with 99% less than 5.2 μm.

Aerosol Characterization It was determined that the DDE of the cyclosporin A powder dried by previous spray was 69.9% ± 7% (n = 10). It was determined that the MMAD was 1.9 with 83% less than 5 μm.

Example 5: Ciclosporin A Acetone Dewdrop At 102-103 ° C With Secondary Drying Preparation of the Solution 1.5 g of cyclosporin A were dissolved in 50 mL of acetone (CLAP grade of J.T. Baker).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Output Temperature 102-103 ° C Input Temperature 140 ° C Feed Speed 5 mL / min Atomizer Flow Rate 15 lit / min Secondary Drying 101 ° C / 5 min.

Once the solution was consumed, the outlet temperature was maintained at 101 ° C for 5 minutes by slowly lowering the inlet temperature to provide secondary drying.

Powder characterization Polarized light microscopy showed that the particles were birefringent. The SEM image of the powder showed that the particles looked round. The DSC scan showed a large isotherm that fluctuated from about 20 ° C to about 70 ° C with a maximum peak at 69 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 123 ° C. The MMD of the powder sample was determined at 1. 20 μm with 87% < 5.2 μm.

Aerosol Characterization It was determined that the DDE of the cyclosporin A powder dried by previous spray was 63.3% ± 7% (n = 10). It was determined that the MMAD was 1.8 μm with 80% less than 5 μm.

Example 6: 90% Cyclosporin A: 10% Citrate Dried by Dew Ethanol At 85 ° C With Secondary Drying Preparation of the Solution 1.35 g of cyclosporin A were dissolved in 50 mL of ethanol (test 200, USP, Spectrum NF grade). 150 mg of Sodium Citrate from Sigma Chemicals were dissolved in 2.5 mL of deionized water. The ethanol / cyclosporin A solution was added to the Sodium Citrate / water solution and stirred rapidly. The resulting suspension is processed in the Dew Dryer.

Spray drying A dry powder comprised of cyclosporin A and sodium citrate (90:10) was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Exit Temperature 85 ° C Inlet Temperature 120 ° C Feed Speed 5 mL / min Atomizer Flow Rate 13 lit / min Secondary Drying 85 ° C / 5 min.

Once the solution was consumed, the outlet temperature was maintained at 85 ° C for 5 minutes by slowly lowering the inlet temperature to provide secondary drying.

Powder Characterization The PXRD of the powder showed that it exhibited a double halo with two maxims 2? equal to 8.5 ° and 17 °. The sharp diffraction peaks that rose above the double halo corresponded to the exploration of sodium citrate dihydrate. Polarized light microscopy showed that the particles were birefringent. The TGA analysis of the powder showed that the powder had 1.3% by weight of residual solvent and a decomposition temperature range of 288-395 ° C. The SEM image of the powder showed that the particles appeared similar to raisins, and some individual facets of citrate crystals were observed. The DSC scan showed a large isotherm that fluctuated from about 20 ° C to about 70 ° C with a maximum peak at 59 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 117 ° C. It was determined that the MMD of the powder sample was 1.11 μm, with 96.7% less than 5.2 μm.

Aerosol Characterization The DDE of the cyclosporin A powder: citrate (90:10) dried by previous spray was determined to be 65.9% ± 5% (n = 10). It was determined that the MMAD was 3.2 with 78% less than 5 μm.

Example 7: Ciclosporin A Ethanol Dewdrops At 85 ° C Without Secondary Drying Preparation of the Solution 1.5 g of cyclosporin A were dissolved in 50 mL of ethanol (test 200, USP, Spectrum NF grade).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Exit Temperature 85 ° C Inlet Temperature 120 ° C Feed Speed 5 mL / min Atomizer Flow Rate 13 lit / min Secondary Drying None Powder Characterization The PXRD of the powder showed that it exhibited a double halo with two maxims 2? equal to 8.5 ° and 17 °. Polarized light microscopy showed that the particles were birefringent. TGA analysis of the powder showed that the powder had 0.7% by weight of residual solvent and a decomposition temperature range of 347-428 ° C. The SEM image of the powder showed that the particles were raisin-like. The DSC scan showed a large isotherm ranging from about 20 ° C to about 70 ° C with a maximum peak at 65 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 122 ° C. It was determined that the MMD of the powder sample was 0.9 μm, with 97.6% less than 5.2 μm.

Characterization of the Aerosol The DDE of the cyclosporin A powder dried by previous spray was determined to be 70.8% ± 3% (n = 10). It was determined that the MMAD was 2.7 with 85% less than 5 μm.

Example 8: Cyclosporin A Spray-dried Ethanol At 101 ° C Without Secondary Drying Preparation of the Solution 1.0 g of cyclosporin A was dissolved in 33 mL of ethanol (CLAP grade).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Output Temperature 101 ° C Input Temperature 138 ° C Feed Speed 5 mL / min Atomizer Flow Rate 14.5 lit / min Secondary Drying None Powder Characterization Polarized light microscopy showed that the particles were birefringent. The SEM image of the powder showed that the particles had slight dimples. The DSC scan showed a large isotherm ranging from about 20 ° C to about 70 ° C with a maximum peak at 68 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 122 ° C. It was determined that the MMD of the powder sample was 2.3 μm, with 86.6% less than 5.2 μm. The powder X-ray diffraction pattern showed a halo with two broad maximums 2? equal to ~ 8.5 ° and ~ 18.8 °. The X-ray pattern of the small angle of the powder showed a peak at 2T equal to 0.2 °, indicative of the two-dimensional order.

Aerosol Characterization It was determined that the DDE of the cyclosporin A powder dried by previous spray was 64% ± 6.9% (n = 7). It was determined that the MMAD was 2.59 with 75% less than 5 μm.

Example 9: Cyclosporin A Acetone Dewdrop At 49 ° C Without Secondary Drying Preparation of the Solution 1.0 g of cyclosporin A was dissolved in 33 mL of acetone (CLAP grade).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Output Temperature 49 ° C Inlet Temperature 60 ° C Feed Speed 5 mL / min Atomizer Flow Rate 14.5 lit / min Secondary Drying None Powder characterization Polarized light microscopy showed that the particles were birefringent. The SEM image of the powder showed that the particles were round and dimpled.

It was determined that the MMD of the powder sample was 3.5 μm, with 70.3% less than 5.2 μm.

Characterization of the Aerosol It was determined that the DDE of the previous spray-dried cyclosporin A powder was from 52. 4% ± 2.1% (n = 7). It was determined that the MMAD was 2.30 with 63.6% less than 5 μm.

Example 10: Ciclosporin A Acetone Dewdrop At 100 ° C Without Secondary Drying Preparation of the Solution 1.0 g of cyclosporin A was dissolved in 33 mL of ethanol (CLAP grade).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Exit Temperature 100 ° C Inlet Temperature 135 ° C Feed Speed 5 mL / min Atomizer Flow Rate 14.5 lit / min Secondary Drying None Powder characterization Polarized light microscopy showed that the particles were birefringent. The SEM image of the powder showed that the particles were round and without dimples. The DSC scan showed a large isotherm that fluctuated from about 20 ° C to about 70 ° C with a maximum peak at 66 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 120 ° C. It was determined that the MMD of the powder sample was 2.75 μm, with 76.3% less than 5.2 μm. The powder X-ray diffraction pattern showed a halo with two broad maximums 2? equal to ~ 7.8.

The X-ray pattern of the small angle of the powder showed a peak at 2? equal to 0.2 °, indicative of the bidimensional order.

Characterization of the Aerosol The efficiency of the released dose (DDE) of the cyclosporin A powder dried by previous spray was determined to be 57.3% ± 3.42% (n = 7). It was determined that the mean aerodynamic diameter of the mass (MMAD) was 2.1 with 70% less than 5 μm.

Example 11: Ciclosporin A Dewdrop Isopropanol At 77 ° C Without Secondary Drying Preparation of the Solution 1.0 g of cyclosporin A was dissolved in 33 L of ethanol (CLAP grade).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Output Temperature 77 ° C Input Temperature 105 ° C Feed Speed 5 mL / min Atomizer Flow Rate 14.5 lit / min Secondary Drying None Powder Characterization Polarized light microscopy showed that the particles were birefringent. The SEM image of the powder showed that the particles were round and dimpled. The DSC scan showed a large isotherm that fluctuated from about 20 ° C to about 70 ° C with a maximum peak at 66 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 122 ° C. It was determined that the MMD of the powder sample was 2.22 μm, with 85.7% less than 5.2 μm. The powder X-ray diffraction pattern showed a halo with two maximum edges 2? equal to ~ 7.5 °. The X-ray pattern of the small angle of the powder showed a peak at 2? equal to 0.1 °, indicative of the bidimensional order.

Characterization of the Aerosol It was determined that the DDE of the cyclosporin A powder dried by previous spray was 69.2% ± 2.42% (n = 7). It was determined that the MMAD was 3.8 with 97.8% less than 5 μm.

Example 12: Ciclosporin A Spray-dried Isopropanol At 104 ° C Without Secondary Drying Preparation of the Solution 1.0 g of cyclosporin A was dissolved in 33 mL of ethanol (CLAP grade).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Exit Temperature 104 ° C Inlet Temperature 145 ° C Feed Speed 5 mL / min Atomizer Flow Rate 14.5 lit / min Secondary Drying None Powder characterization Polarized light microscopy showed that the particles were birefringent. The SEM image of the powder showed that the particles were round and without dimples. The DSC scan showed a large isotherm that fluctuated from about 20 ° C to about 70 ° C with a maximum peak at 66 ° C. The endotherm similar to Tg, which is a fusion, appeared in the scan at an initial temperature of 121 ° C. It was determined that the MMD of the powder sample was 2.36 μm, with 89.9% less than 5.2 μm. The powder X-ray diffraction pattern showed a halo with two maximum edges 2? equal to ~ 8.9 ° and -19 °. The X-ray pattern of the small angle of the powder showed a peak at 2? equal to 0.1 °, indicative of the bidimensional order.

Characterization of the Aerosol It was determined that the DDE of the cyclosporin A powder dried by previous spray was 67.1% ± 2.85% (n = 7). It was determined that the MMAD was 2.7 with 76.8% less than 5 μm.

Example 13: Cyclosporin A Methanol Dewdrop At 63 ° C Without Secondary Drying Preparation of the Solution 1.0 g of cyclosporin A was dissolved in 33 mL of ethanol (CLAP grade).

Spray drying A dry powder comprised of cyclosporin A was produced by spray drying the organic solution using a Buchi Laboratory Spray Dryer B-190 with N2 ATM < 5% of 02 with the following parameters: Exit Temperature 63 ° C Inlet Temperature 88 ° C Feed Speed 5 mL / min Atomizer Flow Rate 14.5 lit / min Secondary Drying None Powder Characterization Polarized light microscopy showed that the particles were birefringent. The SEM image of the powder showed that the particles had many dimples. It was determined that the MMD of the powder sample was 2.37 with 90.1% less than 5.2 μm. The DSC scan showed a large isotherm that fluctuated from approximately 20-70 ° C with a maximum peak at 52 ° C. The endotherm similar to Tg, which is a fusion, appeared in exploration at an initial temperature of 119 ° C.

Aerosol Characterization It was determined that the DDE of the cyclosporin A powder dried by previous spray was 67.2% ± 3.43% (n = 6). It was determined that the MMAD was 2.5 with 80.7% less than 5 μm.

Example 14: The method of Example 1 was followed except that the solution was atomized using a commercially available, standard Buchi nozzle. The average mass diameter (MMD) of the drops using this nozzle was between 7 and 15 μm.

Chemical Stability The analysis by CLAP showed no appreciable degradation of samples of spray-dried cyclosporin powder. Figure 7 is a chromatogram of the spray dried sample reconstituted in a mobile phase after being stored for 15 months at room temperature. It was determined that the powder was chemically stable for the duration of the study.

Example 15: Processing of Cyclosporin A Powder Formulations A Spray-dried Spray-dried cyclosporin A powders were produced from various solvents using a variety of spray-drying temperatures, or without secondary drying. Secondary drying does not seem to have an effect on the properties of the particle. The measurement of the residual solvent showed that the solvent was left in the particles. Lower levels of residual solvent were preferred to minimize any possible lung irritation caused by the solvent. The percent yield, defined as the weight of powder containing CsA recovered in the spray-drying collector divided by the weight of CsA (and any excipient) in the solution that was spray-dried (100% times), fluctuates from the 22% to 78%. Yields of at least about 20% are preferred, with higher yields generally being more preferred, as long as other powder characteristics such as MMD and DDE are acceptable. The fraction of fine particles (%), defined as DDE times the percent < 5μm, fluctuates from 33.3 to 67.7%. Powders with a particle fraction of at least about 25% are preferred. Several lots of powders were spray-dried Ethanol CsA at 70 ° C without secondary drying. The results of those batches are presented in Table 2.

Table 2. CsA Powders Dried by Ethanol Spray at 70 ° C without Secondary Drying Modification of the modes described above to carry out the various embodiments of this invention will be apparent to those skilled in the art following the teachings of this invention as set forth herein. The examples described above are not limiting, but only exemplary of this invention, the scope of which is defined by the following claims. The description of each publication, patent or patent application mentioned in this specification is hereby incorporated by reference in the same degree as if each publication, patent or individual patent application was specifically and individually indicated as incorporated herein by reference. It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention.

Claims (21)

1. CLAIMS Having described the invention as above, the content of the following claims is claimed as property. 1. A cyclosporine, characterized because it is in liquid crystalline form.
2. 2. The cyclosporin according to claim 1, characterized in that the cyclosporin is cyclosporin A.
3. 3. The cyclosporin according to claim 1, characterized in that it shows sharp peaks by X-ray diffraction at a small angle.
4. 4. The cyclosporin according to claim 1, characterized in that it is in the form of a dispersible powder.
5. 5. Cyclosporin according to claim 1, for use as an immunosuppressant, anti-inflammatory or anti-asthmatic agent.
6. 6. Cyclosporin according to claim 1, characterized in that it is prepared by spray-drying a solvent.
7. 7. A composition for pulmonary delivery or administration, characterized in that it comprises liquid crystalline cyclosporine in respirable dust particles.
8. 8. The composition according to claim 7, characterized in that the cyclosporin is cyclosporin A.
9. 9. The composition according to claim 7, characterized in that it is dispersible.
10. 10. The composition according to claim 7, characterized in that it further comprises a pharmaceutically acceptable carrier excipient.
11. 11. The composition according to claim 7, characterized in that it is prepared by spray drying. The composition according to claim 7, characterized in that the cyclosporin comprises at least about 40% by weight of the composition. The composition according to claim 7, characterized in that the particles in the powder have a particle size range with MMD of 0.1 and 15 μm. The composition according to claim 7, characterized in that the particles have a MMAD of less than about 5 μm. 15. The composition according to claim 7, characterized in that it has a distributed dose efficiency of at least about 30%. 16. The method for preparing the composition according to claim 7, characterized in that it comprises: a) mixing cyclosporin with a solvent to form a solution or suspension; and b) spray drying the mixture formed in step a) under conditions which provide a respirable dust. 17. The method according to claim 16, characterized in that it further comprises the step of adding a pharmaceutically acceptable carrier excipient before spray drying. 18. The method according to claim 17, characterized in that the solvent comprises a solution of less than about 50% water. The method according to claim 16, characterized in that the solvent is selected from the group consisting of ethanol, acetone, acetonitrile, isopropanol and methanol. 20. A method for treating or preventing a condition in a subject, which can be prevented or alleviated by cyclosporin, the method is characterized in that it comprises pulmonary administering a therapeutically effective amount of the composition according to claim 7 to a susceptible subject ao who suffers from such condition. The method according to claim 20, characterized in that the condition is selected from the group consisting of asthma, rejection of transplants, sarcoidosis, chronic pulmonary inflammatory disease, chronic obstructive pulmonary disease, emphysema, primary and secondary pulmonary hypertension, cystic fibrosis , pulmonary infections, rheumatoid arthritis and idiopathic pulmonary fibrosis.