EA003476B1 - Dry powder active agent pulmonary delivery - Google Patents

Abstract

1. Particles for delivery of an active agent to the alveoli of a human patient, said particles comprising the active agent and a hygroscopic growth inhibitor, wherein the hygroscopic growth inhibitor is incorporated within the particles in an amount not less than 40% and selected from the group consisting of beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, sulfobutylether-beta-cyclodextrin, dextranomer, hydroxyethylstarch, and maltodextrin, and wherein the particles exhibit a drop in emitted dose under simulated lung conditions of no more than about 25%. 2. The particles of claim l, wherein the hygroscopic growth inhibitor is selected from the group consisting of hydroxyethylstarch, dextranomer and maltodextrins. 3. The particles of claim l, wherein the cyclodextrin is used as the hygroscopic growth inhibitor selected from the group consisting of beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, sulfobutylether beta-cyclodextrin. 4. The particles of claim l, wherein the hydroxyethylstarch is used as the hygroscopic growth inhibitor. 5. The particles of any of claims 1 to 4, wherein the hygroscopic growth inhibitor is present in the powder at an amount sufficient for the powder to exhibit a rate of moisture uptake of no greater than about 0.50% as a function of relative humidity 6. The particles of any of claims 1 to 5, wherein the hygroscopic growth inhibitor is present in the powder in an amount sufficient for the powder to exhibit an overall extent of water uptake of no greater than about 30 weight percent at a relative humidity of 80%. 7. The particles of any of claims 1 to 6, having an emitted dose under ambient conditions of at least 60%. 8. The particles of any of claims 1 to 7, containing from about 40 percent to about 99 percent by weight hygroscopic growth inhibiting agent. 9. The particles of any of claims 1 to 8, which when delivered pulmonarily, are deposited in the deep lung to an extent greater than 20% of the nominal dose. 10. The particles of any of claims 1 to 9, wherein said hygroscopic growth inhibitor is effective to increase the bioavailability of said active agent when delivered to the lung by at least 5%, when compared to the bioavailability observed for the active agent contained in the same particles absent said hygroscopic growth inhibitor and delivered to the lung. 11. Particles for delivery of an active agent to the alveoli of a human patient, said particles comprising the active agent and a hygroscopic growth inhibitor incorporated within the particles in an amount not less than 40%, wherein the particles have a sorption index of less than about 6.5 and the hygroscopic growth inhibitor is selected from the group consisting of beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, sulfobutylether beta-cyclodextrin, dextranomer, hydroxyethylstarch, and maltodextrin. 12. The particles of claim l1, wherein the hygroscopic growth inhibitor is selected from the group consisting of hydroxyethylstarch, dextranomer and maltodextrins. 13. The particles of claim l1, wherein the hydroxyethylstarch is used as the hygroscopic growth inhibitor. 14. The particles of any of claims 11 to 13, having an emitted dose under ambient conditions of at least 60%. 15. The particles of any of claims 11 to 14, containing from about 40 percent to about 99 percent by weight hygroscopic growth inhibiting agent. 16. Particles for delivery of an active agent to the alveoli of a human patient, said particles comprising the active agent and a hygroscopic growth inhibitor, wherein the hygroscopic growth inhibitor is incorporated within the particles in an amount not less than 40% and selected from the group consisting of beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, sulfobutylether beta-cyclodextrin, dextranomer, hydroxyethylstarch, and maltodextrin, wherein the particles maintain an aerosol particle size distribution below 3 microns MMAD when delivered to the alveoli. 17. A method for preparing particles for delivery of an active agent to the alveoli of a human patient, comprising: preparing a mixture of a hygroscopic growth inhibitor selected from the group consisting of beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, sulfobutylether beta-cyclodextrin, dextranomer, hydroxyethylstarch, and maltodextrin, an active agent, and a solvent, wherein the amount of the inhibitor is at least 40% of the total powder weight; and spray drying the mixture to obtain homogenous particles of the hygroscopic agent and the active agent; wherein the particles exhibit a drop in emitted dose under simulated lung conditions of no more than about 25%. 18. The method of claim 17, wherein the hygroscopic growth inhibitor is selected from the group consisting of beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, sulfobutylether beta-cyclodextrin. 19. The method of claim l7, wherein the hydroxyethylstarch is used as the hygroscopic growth inhibitor. 20. The method of any of claims 17 to 19, where the solvent is water. 21. The method of any of claims 17 to 20, wherein the homogeneous particles contain from about 40 percent to about 99 percent by weight hygroscopic growth inhibiting agent. 22. Powder prepared by the method of claim 17. 23. The powder of any of claims 1 to 16 in an aerosol form for delivering the active agent to the lungs of a human patient. 24. The powder of any of claims 1 to 16 for delivering the active agent to the lungs of a human patient by means of a dry powder inhaler.

Description

The present invention relates to improving the delivery of the composition of the active substance in the form of a dry powder to the lungs, and more specifically, the present invention relates to the conversion of dry powder particles into an aerosol, which allows these particles to have hygroscopic growth after inhalation. This characteristic of the powder (i.e., resistance to hygroscopic growth) allows a greater proportion of particles to reach the deep lungs after inhalation, as a result of which the bioavailability of the active substance that enters the lungs increases.

It has already been shown that the delivery of active substances to the lungs is an effective way of prescribing medications for both local and systemic use. The pulmonary formulations of the active ingredient are intended to be delivered by the patient inhaling the dispersion of the drug, so that the active substance contained in the dispersion can reach the lungs. It has already been discovered that some medicinal substances delivered to the lungs are easily absorbed (pass) through the alveolar region directly into the bloodstream. However, the percentage of drug substance after inhalation, which actually reaches the deep lungs, is quite small. With pulmonary delivery, the loss of the drug in the device (in the device for inhalation) is about 30% and about 35% in the oral part of the pharynx (in the upper respiratory tract). Of the remaining 35%, about 20% of the drug substance is lost in the respiratory tract, while about 15% is absorbed in the alveolar region. As stated in the publication of Oopba et al. StSsa1 Reuai \\ '5 ί Tketareiis Aegis Satlet 8uk! substances in the distal airways and alveoli are faster than in the upper airways, since diffusion barriers in these areas are thinner and the surface area is larger. However, since in reality only a small part of the medicinal substance after inhalation reaches the alveolar surface, new approaches are required to increase the amount of the medicinal substance, which ultimately reaches the large circulation.

In accordance with one of the already well-known approaches to resolving this problem, US Patent No. 5,506,203 describes the use of permeability enhancement agents that increase the absorption through the epithelial cell layer in the lower respiratory tract, thereby ultimately increasing the amount of drug substance reaching a large circle of blood circulation. In accordance with this patent, compounds in the form of particles with a diameter of less than 10 microns are used for inhalation. Surfactants, salts of fatty acids, salts of bile, as well as other substances and their derivatives are used as means of increasing permeability. US Patent No. 5,451,569 also describes the use of a pulmonary surfactant designed to enhance the absorption of proteins and peptides in the lungs.

As an attempt to eliminate the need for permeability enhancement, XV O 96/32149 describes pulmonary delivery of aerosolized medicinal substances in the form of a dispersible dry powder. Such drugs are easily absorbed in the lungs, without the need for permeability enhancement agents. Similar efforts to increase the bioaccumulation of medicinal substances after inhalation are described in publication XV O 97/44013. This publication uses aerodynamically light particles (having a density of less than 0.4 g / cm ³ , with an average diameter greater than 5 microns) to improve the delivery of therapeutic or diagnostic agents (substances) to the alveolar region of the lungs. In the publication XVО 98/31346 for an additional increase in the bioaccumulation of medicinal substances it is proposed to introduce a surface-active substance into aerodynamically light particles, which, besides the increase in bioaccumulation, enhances the absorption of medicinal substances.

In addition to the problem of low absorption of drugs with pulmonary delivery through the epithelial cell layer in the lower respiratory tract, there is another factor that reduces the amount of drug after inhalation that reaches the deep lungs, namely hygroscopic growth. Due to its natural solubility in water, the use of most aerosols results in increased sedimentation of particles in the upper respiratory tract as a result of hygroscopic growth (see Nukeu et al. Bulletin 1 Rcagtaséybess1 8c1epsek Volume 79, Number 11, pp. 1009-1014). To study the rate of increase in size (growth) of powders in a humid environment, powders of disodium fluorescein coated with fatty acid were prepared using absorption coacervation technology. The coated powders had ΜΜΑΌ (average aerodynamic particle diameters by mass) in the range of approximately 4 to 7 μm and had a lower growth rate compared to uncoated powders.

Despite the achievements mentioned above, the percentage of drug that actually reaches the alveolar surface of the lungs after inhalation is still quite small. Thus, new approaches are needed to increase the amount of drug substance after inhalation (inhalation) deposited in the deep lungs, so as to increase the bioavailability of drugs after inhalation.

Not only the size and density of particles are important parameters for increasing the bioaccumulation of medicinal substances entering the alveolar region of the lungs, but also their ability to absorb water as they travel through the lungs to the alveoli. The authors of the present invention found that simply coating the particles is not sufficient to minimize water absorption in the lungs, but is required to impart hygroscopic growth properties to all particles in order to maintain an appropriate particle size distribution in aerosols when passing through the lungs to the alveolar surface , without premature deposition in the upper areas of the lungs.

In accordance with the first aspect of the present invention, it is directed to the creation of particles for the delivery of the active substance to the alveoli of patients (people). Such particles contain active ingredient and inhibitor of hygroscopic growth. A hygroscopic growth inhibitor is introduced into the material of particles, and the particle size distribution in aerosols is maintained at less than 3 μm ΜΜΑΌ when it enters the alveoli.

In accordance with another aspect of the present invention, it is directed to the creation of particles that contain the active ingredient and hygroscopic growth inhibitor, and these particles are highly dispersible and provide a drop in the transmitted dose when simulating conditions in the lungs, not exceeding approximately 25%.

In accordance with another aspect of the present invention, it is directed to the creation of particles that have low moisture absorption coefficients. The particles contain the active substance and the inhibitor of hygroscopic growth, and they are additionally characterized by the fact that they have a sorption index of approximately less than 6.5.

In accordance with another aspect of the present invention, it is aimed at creating a method for preparing particles for delivering the active substance to the patient's alveoli. This method involves the preparation of a mixture of hygroscopic growth inhibitor, active ingredient and solvent. The mixture is then spray dried to obtain homogeneous particles containing hygroscopic growth inhibitor and active ingredient. The resulting particles retain a particle size distribution of less than 3 microns in the deep lungs, if delivery is by inhalation. Alternatively, the obtained particles provide a drop in the transmitted dose in terms of imitation of the lungs, not exceeding approximately 25%. In accordance with another alternative, the particles obtained after spray drying have a sorption index of approximately less than 6.5.

In accordance with another aspect of the present invention, it is aimed at creating a method of delivering the active substance to the patient’s lungs, and the aerosol particles, which have the above characteristics here, are introduced due to their inhalation by the patient (during inhalation).

In accordance with another aspect of the present invention, it is aimed at creating a method for increasing the amount of active substance after inhalation deposited in the deep lungs. This method involves the introduction in intended for inhalation of dry powder particles containing the active substance, means of inhibiting (inhibitor) hygroscopic growth, due to which, after becoming an aerosol and inhalation of particles, at least 20% of the nominal dose is deposited in the deep lungs.

The above and other features of the invention will be more apparent from the subsequent detailed description, given as an example, given with reference to the accompanying drawings and examples.

FIG. 1 shows the sorption profiles (absorption) of moisture by various compositions of powders obtained after spray drying, with moisture absorption (wt.%) Being plotted along the vertical axis, and% relative humidity (HH) being plotted along the horizontal axis. (Circles: 20% insulin, 59% sodium citrate, 18% mannitol, 2.6% glycerol; squares: 100% dextran (10K); diamonds: 100% hydroxypropylmethylcellulose; crosses: 100% hydroxypropyl-in-cyclodextrin, and also : 100% low molecular weight hydroxyethyl starch);

FIG. 2 shows the moisture sorption profiles for three different powder formulations obtained after spray drying. (Circles: 20% insulin, 59% sodium citrate, 18% mannitol, 2.6% glycerol; squares: 20% insulin, 2.6% glycerol, 40% hydroxyethyl starch, 18% mannitol, 19% sodium citrate; and diamonds: 100% hydroxyethyl starch). Adding one or more hygroscopic growth inhibitors (HC1) to a particular formulation reduces its moisture absorption.

FIG. Figure 3 shows the results from the TAM recorder (the recorder of the level of activity induced by thermal neutrons) for various insulin compositions in the form of dry powders, which testify to the effectiveness of two typical means of inhibiting hygroscopic growth, leading to a significant decrease in the hydration properties of these powders.

FIG. 4 shows a graph of moisture sorption for three different powder formulations obtained after spray drying, indicating the effectiveness of using formulations containing hygroscopic growth inhibitors to reduce both the water absorption rate (water absorption) and its total absorbed amount. (Circles: 20% insulin, 59% sodium citrate, 18% mannitol, 2.6% glycerol; squares: 100% obtained by spray drying hydroxypropyl-in-cyclodextrin; and diamonds: 20% insulin, 20% leucine, 50% β-cyclodextrin sulfonylbutyl ether, 10% sodium citrate.

FIG. 5 shows a graph of moisture sorption, allowing comparison of 5 different compositions of powders obtained after spray drying. This graph further illustrates the ability of formulations containing hygroscopic growth inhibitors (HC1) to provide a significant reduction in both the rate of water absorption and the degree of its absorption, compared to formulations that do not contain HC1. (Circles: 20% insulin, 20% leucine, 50% hydroxyethyl starch, 10% sodium citrate; squares: 20% insulin, 5% leucine, 50% hydroxyethyl starch, 25% sodium citrate; diamonds: 100% hydroxyethyl starch; crosses: 20% insulin , 59% sodium citrate, 18% mannitol, 2.6% glycerol; ±: 20% leucine, 50% hydroxyethyl starch, 30% sodium citrate).

In accordance with the present invention, a powder composition and a method of pulmonary delivery of particles containing the active substance and means of inhibiting hygroscopic growth are proposed, and the size distribution (particle size distribution) of the particles is maintained at less than 3 μm ΜΜΆΌ when delivered to the alveoli. The present invention surprisingly allows you to provide increased bioavailability of the active substance compared with particles with the active substance, in which there is no inhibitor of hygroscopic growth, or compared with particles that have an inhibitor of hygroscopic growth, adsorbed only on their surface. Due to the presence of a hygroscopic growth inhibitor distributed over the entire volume of the particles, and not over their surface as a coating, as is currently practiced, when the surface of the particles is eroded (eroded) or dissolves when passing through the respiratory tract, new inner layers of the means of inhibition of hygroscopic growth, due to which the particles retain the property of inhibition of hygroscopic growth.

I. Definitions

Used in the text of the description of the invention, the following terms have the following values:

The term active ingredient includes a substance, drug, compound, substance composition, or mixture that provides some pharmacological and often beneficial effects that can be demonstrated on living objects (ίη νίνο) or in vitro (ίη νίΐτο). This includes food, food supplements, nutrients, drugs, vaccines, vitamins and other substances that have beneficial properties. Used in the text of the description, the term additionally includes any physiologically or pharmacologically active substance that provides a local or overall positive effect on the patient.

The term dry powder refers to a composition of such a powder that contains finely divided particles that have free flow and that can be (0 easily dispersed in the inhalation device and (ίί) can be inhaled by the patient, so a portion of the particles reach the lungs and can penetrate into the alveoli. Such a powder is considered suitable for inhalation or suitable for pulmonary delivery. A dry powder typically contains less than about 10% moisture, preferably less than 5% moisture, and even better, less than about 3% moisture.

A hygroscopic growth inhibitor, (HC1) means any material that, when introduced into particles in accordance with the present invention, reduces the rate and / or extent of water absorption. Materials that are used as hygroscopic growth inhibitors are effective if, after they are introduced into the particles at the right concentration, they inhibit hygroscopic growth, under conditions that correspond to typical conditions in the lungs, at least 5%, mainly at least 10%, and even better, at least 15% better compared to particles with the same relative proportions of the components of the particles, but not having HC1.

Hygroscopic growth of particles is usually expressed in terms of the ratio of hygroscopic growth, namely, the ratio of ΜΜΆΌ particles under typical conditions in the lungs to ΜΜΆΌ dry particles before they are inhaled (inhaled). As an example, we point out that particles with a hygroscopic growth ratio of 1 do not change size after inhalation and exposure to conditions present in the lungs. Hygroscopic growth of particles is determined experimentally when powders are placed in

Ί an artificial climate chamber in which conditions existing in the lungs are simulated, namely, a temperature of 32-37 ° C and a relative humidity of 95-99.5%. More specifically, the dose of powders is converted into an aerosol in a growth chamber, after which the aerosol is passed into a cascade hammer crusher to determine the average aerodynamic diameter of particles by mass (ΜΜΑΌ).

Alternatively, it is possible to calculate ΜΜΑΌ a specific powder composition by simulating lung conditions to determine an equilibrium growth ratio. The value ΜΜΑΌ for particles of the powder aerosols in the lungs is determined by calculating the concentration of solids (powder to water ratio), at which the aqueous solution of the powder becomes isotonic, that is, it has a concentration at which a drop of liquid reaches equilibrium in the lungs, which then makes it possible to calculate ΜΜΑΌ for isotonic drop. After that, ΜΜΑΌ for an isotonic drop should be divided into experimentally determined of the powder under ambient conditions, as a result of which a hygroscopic growth ratio is obtained.

In accordance with the present invention, particles containing HC1 and having ΜΜΑΌ less than 3 microns under simulated conditions in the lungs are used.

The simulated conditions in the lungs correspond to a temperature of 32-37 ° C and a relative humidity of 95-99.5%.

The sorption index (31) is the sum of the percent increase in the weight of the dry powder in accordance with the present invention, determined at relative humidities of 10, 20, 30 and 40% (25 ° C) divided by four. The sorption index is determined using a gravimetric sorption analyzer, such as an analyzer of the Όνδ-1000 type, manufactured by ΜοΙδΙιιΐΌ Μеа8 и ^ εηΐ8 8у§1ш (London, United Kingdom), or using a lever scale with a scale calibrated in percent of humidity, produced by our unit. (H 1аеаН (Hialea), Finland).

By active substance particles is meant an active substance, as defined here above, in the form of particles suitable for delivery to the lungs. The particles form a dry powder. It should be borne in mind that the composition of the active substance aerosols can be entered several active substances, so that the use of the word substance (in the singular) does not in any way preclude the use of two or more such substances.

Particles that have a hygroscopic growth inhibitor (HC1) introduced into them are those particles that have HC1 distributed throughout the entire volume of the particle, and not just present as a coating on their surface.

The term pulmonary bioaccumulation refers to the amount of active substance that after its deposition in the lungs has been absorbed and has entered the mammalian circulation, relative to the amount that is absorbed into the blood from the place of subcutaneous injection (% absorbed /% deposited). Representative (representative) model systems for determining pulmonary bioavailability are rats, dogs and primates. Relative pulmonary bioaccumulation can be based on direct injection (drug substance) into the trachea or inhalation.

The term Released dose (ΕΌ) gives an indication of the distribution of the dry powder in an appropriate device for inhalation after it has been activated or when dispersed. ΕΌ is defined as the ratio of the released dose to the nominal dose (that is, the mass of powder needed to form a single dose placed in an appropriate device for inhalation before it triggers). ΕΌ is a parameter that is found experimentally, usually in a test tube (ίη-νίΐτο) with a simulated patient dose. To determine the value of ΕΌ, the nominal dose of dry powder is placed in an appropriate device for inhalation, which is then triggered and sprays the powder. The resulting cloud of aerosols is drawn in using a vacuum through a calibrated filter connected to the mouthpiece of the inhalation device. The amount of powder that reaches the filter forms the released dose. For example, if from a 5 mg dose of a dry powder placed in an inhalation device, 4 mg of powder is precipitated on a specified calibrated filter, then the released dose for a given dry powder formulation is: 4 mg (released dose) / 5 mg (nominal dose) = 80%. For inhomogeneous powders, the ΕΌ values give an indication rather of the distribution of the medicinal substance in the device for inhalation after it is triggered, rather than the distribution of the dry powder, and these values are based on the weight of the medicinal substance rather than on the total weight of the dry powder.

The term “loss of the released dose” when simulating conditions in the lungs means the difference between the value of ΕΌ in ambient conditions (%) and the value of ΕΌ at a temperature of 3237 ° C and at a relative humidity of 95-99.5%.

Dispersible powder refers to such a powder, which has a value of рав equal to at least about 30%, preferably 40-50%, and even better, at least about 50-60%.

The average particle diameter by mass or ΜΜΌ is used to measure the average particle size, since the powders in accordance with the present invention are typically polydisperse (i.e., contain a range of particle sizes). The values of ΜΜΌ are usually found using centrifugal sedimentation (sedimentation), but other known techniques can be used to determine the average particle size (for example, electron microscopy, light scattering, laser diffraction).

The average aerodynamic particle diameter by mass or ΜΜΑΌ is used to measure the aerodynamic size of the dispersed particle. The aerodynamic diameter, which is used in the description of the powder in aerosols to determine its behavior during the deposition, is the diameter of the sphere of unit density, which has the same deposition rate in air as the particle. The aerodynamic diameter takes into account the shape of the particle, as well as its density and physical size. As used herein, the term refers to the midpoint or median of the size distribution of aerodynamic powder particles in aerosols, with the particles obtained using a cascade hammer mill, unless specifically indicated otherwise.

A pharmaceutically acceptable excipient or carrier is an excipient that can be incorporated into the particles in accordance with the present invention and can enter the lungs along with the particles without creating significant harmful toxicological effects for the patient, especially for his lungs.

A pharmaceutically effective amount or a physiologically effective amount of the active substance is the amount of the active substance present in the dry powder composition described above, which is necessary to achieve the desired level of the active substance in the patient’s bloodstream, allowing the expected physiological response to be obtained after pulmonary administration of such a composition. The exact amount depends on many factors, for example, the type of active ingredient, the specific activity of the composition, the specific delivery device used, the physical characteristics of the powder, the type of its use and the patient's data, and this amount can be easily determined by a specialist based on the information available here .

Ii. Inhaled Powder Components

Particles according to the present invention are formulated so as to be resistant to hygroscopic growth, which normally occurs upon pulmonary administration of powder formulations, and to allow a greater proportion of particles to reach deep lungs after inhalation. This particle characteristic, i.e., resistance to hygroscopic growth, is achieved by introducing a hygroscopic growth inhibitor, i.e., such a means, the presence of which in the particles effectively reduces the rate and / or degree of water absorption by the particles, especially when exposed to environmental conditions lungs.

A. Active ingredient

The active ingredient, which is intended for introduction into the powder compositions described herein, includes antibiotics, antiviral agents, anepileptics, analgesics, anti-inflammatory drugs and bronchodilators. The active substance can be used inorganic or organic compound, including, but without limitation, medicinal substances that affect the peripheral nerves, adrenoreceptors, cholinergic receptors, skeletal muscles, the cardiovascular system, smooth muscles, the circulatory system, synoptic centers, neuroeffector synapse (connective) centers, endocrine and hormonal systems, the immune system, the reproduction system, the skeletal system, the autocoid system, the system nutrition and the excretory system, as well as the histamine system of the central nervous system. Suitable active substances may be selected, e.g., among polysaccharides, steroid agents, hypnotics and sedatives, antidepressants, tranquilizers, anticonvulsants, muscle relaxants and miokontraktantov, antiparkinsonizma agents, analgesics, anti-inflammatory agents, antimicrobial and anti-malarial agents; hormonal drugs, including contraceptives; sympathomimetic agents, polypeptides and proteins capable of causing physiological effects, diuretics, means for regulating lipids, anti-androgenic agents, anti-parasitic agents, tumor and antitumor agents, agents against hypoglycemia, nutritional and nutritional supplements, growth supplements, fats, anti-enteriters, electrolytes, vaccines and diagnostic tools.

Examples of active ingredients suitable for use in accordance with the present invention include, but are not limited to, calcitonin, erythropoietin (EPO), Factor VIII, Factor IX, ceredase, cerezim, cyclosporine, granulocyte colony stimulating factor (CC8E), alpha inhibitor -1 proteinase, elkatonin, granulocyte macrophage colony stimulating factor (СΜС8Ε), growth hormone, human growth hormone (HCH); growth hormone releasing hormone (SNYAN);

heparin, low molecular weight heparin (LMH), interferon-alpha, interferon-beta, interferon-gamma, interleukin-2; hormone releasing luteinizing hormone (LNH); insulin, somatostatin; somatostatin analogs, including octreotide; vasopressin analogue, follicle stimulation hormone (Ε8), insulin-like growth factor, insulintropin, interleukin-1 receptor antagonist, interleukin-3, interleukin-4, interleukin-6, macrophage colony stimulating factor (MC8E), nerve growth factor, parathyroid hormone ( PTH), thymosin alpha 1, Pb / Sha inhibitor, alpha-1 antitrypsin, UYA-4, respiratory syncytial viral antibody, gene of cystic fibrous transmembrane regulator (NET), deoxyribonuclease (Opake), bactericidal permeability enhancement protein (ΒΡΙ), SMU antibody, re apepslc, ancillary interferon-a-1, 13-C1K retinoic acid; In addition, the active substances may additionally contain nucleic acids present in the form of simple nucleic acid molecules, viral vectors, interacting viral particles, plasmid DNA or RNA, or other nucleic acid constructs of the type suitable for transfection or transformation of cells, in particular cells of the alveolar region of the lungs. The active substances can have various forms, such as soluble and insoluble in water, saturated and unsaturated molecules, as well as components of complexes of molecules or pharmacologically acceptable salts. Active substances can be in the form of naturally occurring molecules, or they can be obtained by recombination, and they can be analogs of naturally occurring or derived by recombination of proteins with one or more amino acids added or removed. Moreover, the active substances may contain killed viruses or viruses with impaired activity, suitable for use as vaccines.

The amount of active ingredient in the aerosol particles should be sufficient to deliver a therapeutically effective amount of the active substance to a single dose, allowing to obtain the desired result. In practice, this amount varies widely depending on the type of substance, its bioactivity, the severity of the disease, the patient population, the requirements for prescribing the dose, as well as the desired therapeutic effect. The particles typically contain from about 1 to about 99% by weight of active ingredient, typically from about 2 to 95% by weight of active ingredient, and preferably from about 5 to 85% by weight of active ingredient. Particularly useful particles that allow you to supply the active ingredients in doses from 0.001 mg per day to 100 mg per day, and mostly from 0.01 mg per day to 50 mg per day.

B. Hygroscopic growth inhibitor

An essential characteristic of particles is the presence of a hygroscopic growth inhibitor. The hygroscopic growth inhibitor (ΗΟΙ) effectively reduces the rate and / or extent of moisture absorption by particles during inhalation, so that the particles retain MMAE less than 3 microns when entering the alveoli.

Material that is suitable for use as ΗΟΙ is primarily selected when preliminarily determining its moisture absorption profile after spray drying; low absorption materials, such as those shown in FIG. 1 are preferred for use in accordance with the present invention. These ΗΟΙ materials are subjected to additional testing for suitability for the preparation of particles that contain appropriate amounts of ΗΟΙ (typically more than about 5 to 10% by weight of the composition). In some cases, ΗΟΙ may be present not only in the bulk of the powder, but also form an additional coating on the surface of the particles. Then, moisture isotherms are found for active substance particles containing ΗΟΙ, as well as for control particles having the same relative amounts of the components, but without ΗΟΙ, to determine if the presence of ΗΟΙ effectively affects the reduction in the rate or rate of water absorption by the powder. Usually, both low and high concentrations ΗΟΙ are checked to determine the range of introduction введения in the powders in accordance with the present invention.

Among the materials that can be used as inhibitors of hygroscopic growth, you can specify (but without limitation) the following: double-chain phospholipids, cyclodextrins and their derivatives, hydroxyethyl starch (ΗΕ8), dextran, dextranomer, maltodextrins, starches, hydroxypropylmethylcellulose (HPMC), ethylhydroxyethylether of cellulose, as well as other derivatives of cellulose, such as those described in the publication Ce11 and Csk: Sjesh1sa1, Vyuyet1sa1 apb Ma1epa1 Akresy (Cellulose: chemical, biochemical and material aspects) (ESH Iopg General 8Spec ίη Ro1teg 8s1epse apb Tesypokoda), 1.E., V.8s. Keppebu, OO, V.8c. Ryshr, R.A. ^ 1 Shatk (editor), as well as in the publication Srggeyeptsue Se11i1oke Siestschyu (Complete chemistry of cellulose), Ό. K1ett (Editor), Weigat RyrShr, T. Yesh / e (1998). In some cases, the active substance itself can act as a hygroscopic growth inhibitor. Among the active substances that tend to perform the functions of HC1, you can specify insulin, salmon calcitonin and PTH.

Among the double-chain phospholipids that can be used in accordance with the present invention, phosphatidylcholines can be mentioned, such as 1,2-dimyristoyls-glycero-3-phosphocholine (IMR), 1,2-dipalmitoyl-ch-glycero-3-phosphocholine (IRRS), 1,2dystearoyl-ns-glycero-3-phosphocholine (IZRS), 1,2-dioleoyl-ns-glycero-3-phosphocholine (IORS), 1-palmitoyl-2-oleoyl-ns-glycero-3-phosphocholine ( RALO), as well as other similar compounds. Also suitable for use as a hygroscopic growth inhibitor are phosphatidylethanolamines, such as 1,2-dimyristoyl-cn-glycero-3-phosphoethanolamine (EMPE). 1,2-dialmitoyl-ns-glycero-3-phosphoethanolamine (ERRE), 1,2-distearoyl-ns-glycero-3-phosphoethanolamine (EZRE), 1,2-dioleoyl-ns-glycero-3phosphoethanolamine (POR), and also derivatives of phosphatidylglycerols and phosphatidic acids.

Cyclodextrins are another class of compounds that can be used as hygroscopic growth inhibitors. Cyclodextrins, which are cyclic oligosaccharides and have the shape of a truncated cone with a hydrophobic cavity in the center, contain more than six glucose residues. Among the cyclodextrins that can be used in accordance with the present invention, you can specify, in accordance with the number of glucose residues, alpha-cyclodextrin (six glucose residues), beta-cyclodextrin (seven glucose residues) and gamma-cyclodextrin (eight glucose residues), as well as derivatives thereof, such as 2-hydroxypropyl-in-cyclodextrin (abbreviated 2-HPC) and β-cyclodextrin sulfonyl butyl ether. 2-HPAc is a particularly preferred filler, as can be seen from its moisture sorption profile (Fig. 1). At a given relative humidity of 80%, 2HPB has a mass change due to moisture absorption, which is only about 16% for about 8 hours. Cyclodextrin has a similar profile. The favorable properties of moisture sorption for an approximate composition, which contains sulfobutyl ether-β cyclodextrin (abbreviated 2-BEAS), are shown in FIG. 4. Thus, these materials are (ί) sufficiently resistant to water absorption and (ίί) have a low water absorption rate, which allows them to be successfully used for incorporation into powders in accordance with the present invention.

Also useful for use as hygroscopic growth inhibitors are dextrans, which are polysaccharides containing glucose monomers. The dextrans used in accordance with the present invention have a molecular weight in the range of from about 10,000 to 100,000. Preferred are dextran 10, dextran 40, dextran 70 and dextran 75. Dextran derivatives, such as dextranomer (dextran 2,3 -dihydroxypropyl 2-hydroxy1,3-propanediyl ether), maltodextran and dextran sulfate sodium. An experiment was conducted to find a balance of moisture sorption to assess the resistance of dextran to moisture absorption, according to which, at a relative humidity of 70% dextran after spray drying, it absorbs only 19% of water, and at a relative humidity of 80% dextran has a water sorption of 24% by weight, as shown in example 3 and in fig. one.

Also useful for use as hygroscopic growth inhibitors are cellulose derivatives, such as hydroxypropylmethylcellulose (HPMC), ethylhydroxyethyl ether cellulose, and hydroxypropylcellulose with a molecular weight in the range from 10,000 to 400,000.

Starch derivatives can also be used as inhibitors of hygroscopic growth. One particularly preferred inhibitor of hygroscopic growth is hydroxyethyl starch (NEZ), which has a molecular weight in the range of from about 70,000 to about 400,000 (see, for example, Fig. 2). The review of NEZ is given in Publication Siena Meb (1999) 25: 258-268.

Also useful for use as a hygroscopic growth inhibitor is maltodextrin, which is a hydrolyzed starch, as well as its commercially available derivatives. Preferred is maltodextrin 40, which has an average molecular weight of about 3,600.

In HC1 inhibitors, which can be successfully used in the particles and methods according to the present invention, effective minimization of hygroscopic growth should be combined (1) with a lack of toxicity at the concentrations used and (2) with good powder properties, i.e. no adhesive or wax (solidifying) consistency in the solid state. The toxicity of this material can be tested using standard methods, such as the MTT assay described, for example, in the publication ΐηΙ. 1. Ryatsh. 65 (1990), p. 249-259.

The hygroscopic growth inhibitor is present in the particles in an amount sufficient to minimize or prevent the hygroscopic growth of the particles, so that the particles retain their size less than 3 μm when aerosolized in the alveoli. The optimal ratio of the active substance to ΗΟΙ can be established for any given ΗΟΙ by checking various properties in the ίη νίίτο model (in vitro). For example, the active ingredient is usually combined with ΗΟΙ, such as hydroxyethyl starch, in the following weight proportions: 10/90, 25/75, 50/50, 75/25 and 90/10, to determine which of them gives a significant reduction in the degree or speed water absorption powders. These data allow us to determine the optimal concentration of ΗΟΙ. Different ΗΟΙ in combination with different active ingredients and with possible additional excipients will have different optimal concentrations, so each ΗΟΙ should be checked separately (in each situation).

Typically, the particles contain at least about 5 to about 20 wt.%, Preferably at least about 20 to about 40 wt.% ΗΟΙ, and even better, at least about 40 to about 60 weight. .% (or more) ΗΟΙ. The amount ΗΟΙ needed to reduce the moisture absorption properties of the powder will be less when the active substance is protein or polypeptide, since the proteins or polypeptides themselves also function as an inhibitor of hygroscopic growth. If the active substance is not a protein or a polypeptide, then the particles should preferably contain at least about 40%, and the amount of ΗΟΙ in the particles should be in the range of approximately 40 to 99% by weight. The presence of ΗΟΙ in particles maximizes the deposition (deposition) of aerosols in the lower respiratory tract, in particular, on the alveolar surface, and not in the mouth, throat or upper respiratory tract, resulting in increased bioavailability of the active substance entering the lungs.

B. Other fillers

In addition to the hygroscopic growth inhibitor, powders with the active ingredient in accordance with the present invention may contain pharmaceutical carriers or excipients suitable for respiratory and pulmonary administration. Such carriers can simply act as ballast when it is desirable to reduce the concentration of the active ingredient in the powder that is assigned to the patient. However, carriers can also serve to further improve the dispersion of the powder in the device for its spraying, which allows for more efficient and reproducible delivery of the active substance and improves the characteristics of the active substance (for example, flowability and consistency), which facilitates the preparation of the powder and filling the device for spraying.

In particular, filler materials can often perform the function of improving the physical and chemical stability of particles, in order to further reduce the residual moisture content and interfere with its absorption, improve particle size distribution, degree of aggregation, surface condition (eg, roughness), ease of inhalation and the direction of the particles in the deep lungs.

These fillers, if any, are usually present in the composition in amounts from about 1 to 50%, and contain (but without limitation) the following substances: proteins, peptides, amino acids and hydrocarbons (for example, sugars, including monosaccharides, di-, tri-, tetra- and oligosaccharides, sugar derivatives such as alditols, aldonic acids, esterified sugars and the like, as well as polysaccharides or sugar polymers), which may be present in the singular or in combinations.

Examples of protein fillers include human serum albumin, such as human serum albumin (Η8Ά), human recombinant albumin (GNA), gelatin, casein, and the like. Examples of representative amino acids and polypeptide components that can also function as a buffer capacity include alanine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame , di-and tripeptides, such as trileucine, and the like. Among the hydrocarbon fillers suitable for use in accordance with the present invention, you can specify, for example, monosaccharides, such as fructose, maltose, galactose, glucose,-mannose, sorbose; disaccharides such as lactose, sucrose, trehalose, cellobiose, and the like; polysaccharides, such as raffinose, melezitoza and the like; and alditols, such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol), and the like.

The composition may also contain a buffer or pH adjusting agent. Representative representative buffers include organic acid salts such as salts of citric acid, ascorbic acid, gluconic acid, carbonic acid, tartaric acid, succinic acid, acetic acid, and phthalic acid; and also tgb. tromethamine hydrochloride or phosphate buffers. In addition, compositions in accordance with the present invention may also contain additional fillers and additives, such as X118 (polymer sugar), flavoring agents, antimicrobial agents, sweeteners, antioxidants, antistatics, surfactants (for example, polyoxyethylene fatty acid esters, such as ΤνΕΕΝ 20 and 'ΤνΕΕΝ 80), as well as chelating agents (such as, for example,). Other pharmaceutical excipients and / or additives suitable for use in the matrix compositions described herein are listed in the Ketshd1op publication: THe 8Aeps & Rtasse οί Ryattas, 19 ^4B ., ^ 1SHT8 & ^ 1SHT8, (1995), and also in Ryushap to Ee ^ to Regegeps, 52 ' ^{1 to} her., Mei1s1 Nsonopks ^, Mopulua, N1 (1998), which are included in this description by reference. Among the preferred excipients suitable for use in the compositions according to the present invention are mannitol, raffinose and sodium citrate, leucine, isoleucine, valine, sucrose, raffinose, tri-leucine and mannitol.

Iii. Preparation of Powder Formulations

The formulations of the active substance in the form of a dry powder are preferably prepared by spray drying, under conditions in which mainly amorphous powder is obtained. Spray drying of the composition is carried out, for example, in accordance with the publication in the publication of the Manual of Spray Drying, 5 ^{4b to} it., K. Mayega, 1о1ш \ Uyeu & 815, 1ps.,, ΝΥ (1991), and in the publication Р1а1х, I. and others., 1paypayopa1 Pa1ep1 Rysayop No. XVO 97/41833 (1997), which are included in this description by reference.

First prepare solutions, emulsions or suspensions that contain the active ingredient, hygroscopic growth inhibitor and other possible fillers. Solutions or suspensions for spray drying typically contain from about 0.1 to about 10 weight percent per volume of solids. The pH range in solutions is usually maintained from about 3 to 9, and these values depend on the degree of influence of pH on the stability of the active substance. More neutral pH values are preferred, since such values can help maintain the physiological compatibility of the powder after it is dissolved in the lungs. The compositions after their preliminary spray drying may also contain additional water-soluble solvents, such as alcohols or acetone. Among representative alcohols, lower alcohols such as methanol, ethanol, propanol, isopropanol, and the like can be mentioned. The resulting solutions, as a rule, contain the active ingredient in a concentration of from 0.01 wt.% (Active ingredient per solution volume) to about 2% (weight per volume), and usually from 0.1 to 1% (weight per volume).

After that, spray-drying the solutions in a conventional spray dryer, for example, in one that can be purchased from Νπ наL / 8 (Netherlands), Visa (Switzerland), or another similar one, resulting in a stable, dry powder. The optimal conditions for spray drying depend on the composition of the components and are usually determined experimentally. Air is usually used as a spray-drying gas, although inert gases such as nitrogen or argon can also be used. The temperature at the inlet and outlet of the gas used to dry the sprayed material is chosen in such a way as to avoid loss of activity and decomposition of the active substance in the specified material. This temperature is usually determined experimentally, but usually the inlet temperature is in the range of approximately 50 to 200 ° C, while the outlet temperature lies in the range of approximately 30 to 150 ° C.

Alternatively, dry powders can be prepared using freeze drying, vacuum drying, spray freeze drying, treatment with supercritical fluid, as well as other types of evaporation drying. In some cases, it is desirable to have a dry powder composition in a form that has improved processing characteristics, for example, reduced electrostatic characteristics, better flowability, low sintering, and the like, which can be achieved by preparing compositions formed from aggregates of small particles, i.e. from the aggregates or agglomerates of the above-mentioned dry powder matrix particles, and these aggregates easily decompose back into the fine powder components necessary for pulmonary dosage avki, as described, e.g., in U.S. Patent № 5,654,007, 1997, the disclosure of which is incorporated herein by reference. Alternatively, powders can be prepared by agglomerating the powder components, screening the materials to obtain agglomerates, spheronizing to obtain more round agglomerates, and sizing by size to obtain a product with uniform sizes, as described, for example, in Aypesk, C., etc., PCT RIUIyop № νθ 95/09616, 1995, which is included in this description by reference. Dry powders can also be prepared by mixing, grinding, sieving, or jet grinding the components of the composition in the form of a dry powder. The resulting powders are usually mainly amorphous.

Dry powders are preferably maintained in a dry state (i.e., at relatively low humidity) during their manufacture, processing and storage.

Iv. Powder characteristics

The powder particles in accordance with the present invention have the ability to maintain an aerodynamic diameter of less than 3 microns when entering the alveoli. As shown in example 1, powders with an initial of 3.5 μm, which do not contain a hygroscopic growth inhibitor, behave similarly to powders having of 5-6 μm. The calculation also shows that in the state of equilibrium in the lungs, ΜΜΆΌ of the powder grows to 9 microns. From the data we can conclude that the introduction of the hygroscopic growth inhibitor powders into the compositions described here effectively reduces the rate and / or the degree of hygroscopic growth of dry powder particles, as a result of which not only increases the bioavailability of the active substance in the powder particles, but also increases the dispersion of these compositions

In powders in accordance with the present invention, ΜΜΆΌ particles in most cases are less than 3 microns before pulmonary delivery. Typically, the particles to some extent increase the size (grow) with pulmonary delivery, but to a lesser extent than in the absence of a hygroscopic growth inhibitor, and usually have a hygroscopic growth ratio of less than 2.5, preferably less than 2.0, preferably from 1.5 to 2.0, and even better, less than 1.5. The relationship of hygroscopic growth can be determined experimentally by comparing ^ 4 ^ _{4AO nt3} , | _eM | for powders in ambient conditions with ΜΜΑΌ _1υη6 for powders when simulating conditions in the lungs in an artificial climate chamber (ΜΜΑϋ ₁ „ _{η £} / ΜΜΑϋ ₃ ^ Η; _{βπ |} ). Alternatively, ΜΜΑΌ for a particle by simulating conditions in the lungs can be calculated as follows. First of all, when using molecular weights of all forming (components) of the particles, isotonicity is determined for each component. Then the resulting isotonicity is added to find the particle isotonic. From this value, determine the volume of fluid required to achieve the specified isotonicity; this volume is then taken as the volume of the sphere. From the indicated volume of the sphere, find the diameter of the sphere, which is the calculated ΜΜΑΌ of particles when simulating conditions in the lungs. This calculated ΜΜΑΌ is then used to determine the ratio of hygroscopic growth as described here earlier.

The characteristics of the absorption of moisture by particles are usually determined in experiments on moisture sorption. Data on moisture sorption for powders can be determined using various techniques, such as moisture sorption balance or a thermal activity monitor (TAM). Moisture sorption balances are determined by measuring the loss or weight gain as a function of the increase in relative humidity at a constant temperature. Transporting gas introduced at a known relative humidity (KH) is created by mixing wet and dry gas streams. This gas then acts on a sample placed in a non-hygroscopic sample cup mounted on a microbalance. Depending on the morphology of the sample, it can absorb, adsorb or desorb moisture. This sorption is detected using a microbalance as an increase or decrease in the weight of the sample. A computer program is used to collect data (such as time, temperature, relative humidity and weight) throughout the experiment, at user-defined equilibrium points.

The powders in accordance with the present invention can also be characterized using the sorption index 81, that is, using the sum of the percent weight gain of the powder, determined at 10, 20, 30 and 40% relative humidity divided by four. The rate of sorption is determined using a gravimetric sorption analyzer, such as an OU8-1000 type analyzer. manufactured by 8GGase 8ea8i ^ eteηΐ8 8ЧЧHst (London, United Kingdom), or with the help of lever scales with a scale graduated in percentage of humidity, produced by UT1 Sogrogabop (H1aayai (Hialea), Finland). Powders in accordance with the present invention typically have sorption rates of approximately less than 7.5, preferably less than 7.0, preferably less than 6.5, and even better, less than 6.0. Powders with such values of sorption 81 are given in example 2.

Preferred for use in accordance with the present invention are powders that absorb water slowly, that is, at a rate of less than about 0.75% moisture as a function of relative humidity, preferably less than about 0.50% moisture as a function of relative humidity, preferably less than approximately 0.35% moisture as a function of relative humidity, and even better, less than approximately 0.25% moisture as a function of relative humidity (see, for example, FIG. 1). In accordance with another method, powders absorb less than approximately 60% moisture (wt.%), Preferably less than 30% moisture, preferably less than 25% moisture, and even better, less than 20% moisture, and at best, approximately from 10 to 20 wt.% water in humid conditions, at a relative humidity of 80%. FIG. 1 and 2 it is shown that powders that contain a hygroscopic growth inhibitor, as compared with compositions of powders that do not contain HOT, have both a reduced rate of water absorption (as evidenced by smaller slopes of curves compared to control compositions), and a lower overall degree water absorption.

When considering FIG. 1, it can be seen that at a relative humidity of 80%, a control powder obtained by spray drying, which contains 20% insulin, 59% sodium citrate, 18% mannitol and 2.6% glycine, absorbs 60% moisture by weight, while as under the same conditions, dextran, hydroxypropyl methylcellulose, hydroxypropyl-in-cyclodextrin and hydroxyethyl starch, obtained by spray drying, absorb 24, 16, 16 and 24% moisture, respectively, which indicates the best hygroscopic growth inhibition properties for these materials. Similarly, as shown in FIG. 2, at a relative humidity of 80%, while the control powder absorbs 60% of moisture, in the same conditions powders obtained by spray drying, which contain 20% insulin, 40% hydroxyethyl starch, 2.6% glycine, 18% mannitol, 19% sodium citrate, and 100% hydroxyethyl starch absorb 50 and 24% moisture, respectively. Moreover, in FIG. 1 and 2, it can be seen that the rate of water absorption is significantly reduced in materials with the presence of HC1 compared with control materials.

The powders in accordance with the present invention can also be characterized as having good dispersion properties in hot and humid environments, such as those found in the lungs. Powders in accordance with the present invention usually have a loss of the released dose (ΕΌ) at a temperature of 32 ° C and at a relative humidity of 95% (compared to ΕΌ in ambient conditions), not exceeding approximately 30% (i.e., EP _at L1f1 minus EO | _{||| t |} , | equals 30 or less), preferably from about 20 to 25%, preferably not more than 15%, and even better, not more than about 10%.

As an illustration, example 2 shows the compositions of powders, the values of which during the assessment in an artificial climate chamber are reduced only by approximately 10-15% (for compositions with 60% maltodextrin). Also, good compositions of им when imitating conditions in the lungs have powder compositions containing 60% hydroxyethyl starch. As can be seen from the table. 1, an increase in the amount of hydroxyethyl starch from 40 to 60% (samples 2/3 in comparison with 4/5) effectively reduces the падение drop in an artificial climate chamber. On average, these compounds have a fall of about 20% when simulating conditions in the lungs, while maintaining the values of ΕΌ at about 55%.

The released dose (ΕΌ) of powders that contain HOT, in environmental conditions, exceeds 30%, and usually exceeds 40%. Advantageously, the dosage of powders released in accordance with the present invention exceeds 50%, and often exceeds 60%. The powders in accordance with the present invention contain a large proportion of aerosol particles with small sizes, and therefore they are extremely effective in delivering them as aerosols, as they (ί) reach the alveolar region of the lungs, () diffuse into the interstitium, and (sh) then penetrate the bloodstream through the endothelium.

Dry powders in accordance with the present invention typically have an average moisture content under ambient conditions of approximately less than 10% by weight, usually approximately less than 5% by weight, and preferably approximately less than 3% by weight. Such low moisture solids are more stable than high moisture solids.

V. delivery of powder to the lungs

The dry powder formulations described herein, containing an HO1 inhibitor, are advantageously delivered using any suitable dry powder inhaler (ΌΡΙ). that is, a device for inhalation, by means of which the patient, while inhaling, receives the medicinal substance in the form of a dry powder, transported to the lungs. Preferred are dry powder inhalation devices that are included in Therapeutic Inhalation Systems, such as those described in US Pat. No. 5,458,135 (1995), US Pat. No. 5,740,794 (1998), and US Pat. No. 5,785,049 (1998).

Upon delivery of the powder using a device of the specified type, the dry powder is stored in a tank having a lid with a punched (punctured) hole or other access surface, and is mainly stored in a blister pack or cartridge, and the tank may contain a single dose or a plurality of unit doses. Known methods of filling a large number of cavities with metered doses of the drug substance in the form of a dry powder are described, for example, in the publication Ratkk, .Τ.

Also suitable for delivering the dry powders described herein are dry powder inhalers of this type, which are described, for example, in US Pat. No. 3,906,950 (1974), as well as in US Pat. No. 4,013,075 (1977), and in these inhalers a pre-measured dose of dry powder is contained in a hard gelatin capsule.

As examples of other devices for spraying powder, intended for delivery of dry powder to the lungs, you can specify the device described in European patent number EP 129985 (1988); in European Patent No. EP 472598 (1996); in the European patent 23 no. EP 467172, (1994), and in US patent No. 5,522,385 (1996). Also suitable for delivering matrix dry powders in accordance with the present invention are inhalation devices such as Akkga-Egaso TiKBiNAEK. A device of this type is described in detail in US Pat. No. 4,668,218 (1987), in US Pat. No. 4,667,668 (1987), and also in US Pat. No. 4.805,811 (1989). In addition, devices are also suitable that use a piston (plunger) to inject air intended either to entrain powdered medicinal substance and remove it from the carrier mesh when air is passed through the mesh, or to mix (air) with the medicinal substance in the mixing chamber. , followed by the introduction of powder to the patient through the mouthpiece of the device, as described, for example, in US patent No. 5,388,572 (1997).

Dry powders containing the HC1 inhibitor can also be delivered using a metered-dose inhaler (ΜΌΙ) with supercharging, which contains a solution or suspension of the drug substance in a pharmaceutically inert liquid propellant, for example, in a chlorofluorocarbon or in a fluorocarbon, as described in US Pat. No. 5,320,094 ( 1994) and in US Pat. No. 5,672,581 (1994).

Before using, the HC1 inhibitor containing dry powders is usually stored at ambient conditions, and preferably at temperatures below about 25 ° C and at relative humidities (EN) in the range from 30 to 60%. Better relative humidity conditions, for example, less than about 30%, can be achieved by using a desiccator in a secondary dose package. Inhalable dry powders in accordance with the present invention are characterized not only by good aerosol properties, but also by good stability.

When converted into an aerosol for direct delivery to the lungs, the active ingredient contained in the dry powder formulations described here provides an increase in the bioaccumulation in the lungs due to the presence of HC1 in the powder particles, which allows a greater proportion of particles to reach deep lungs after inhalation, without premature deposition in the upper respiratory tract due to hygroscopic growth. Thus, these dry powder formulations containing HC1 make it possible to prescribe smaller amounts of the drug per unit dose and may even eliminate the need for several inhalations per day. Moreover, the presence of HC1 provides increased stability of the powder by reducing or eliminating the absorption of moisture, resulting in increased shelf life during storage and stability during transportation of dry powder formulations.

All publications, patents or patent applications mentioned in the description of the invention are included in this description by reference to the extent that each individual publication, patent or patent application has.

The following are examples that have no character limiting the scope of patent claims.

Examples

Materials and methods

Calcitonin salmon was purchased at the company Vasiyesh (USA, Toggaps, CA).

Human serum albumin (H8A) was purchased from the company Μ Peck 1ps. (USA, Kapkakee, 1b). Sodium citrate sodium dihydrate was obtained from 1.T. Wakeg (United States, Ryshk, N1). B, adipalmitoylphosphatidylcholine (ERRS) was obtained at the company Auapy Rotif b1p1bk (United States, Alabata).

Example 1

To study the absorption of moisture and the properties of hygroscopic growth, the following particles (powder), which contain the active substance, were prepared.

A. Preparation of powder

Calcitonin salmon powders were prepared as follows. The mass of calcitonin salmon (kSa1yoshp) was dissolved in buffered sodium citrate containing mannitol and H8A, and an aqueous solution was obtained that contains a final solids concentration of 7.5 mg / ml and has a pH of 6.7 ± 0.3. After that, the solution was filtered through a 0.22 μm filter, and then spray-dried using a spray dryer like VisA 190 (Visa BajoiesPshk AS, Meterkeddk1gakks, Switzerland). The spray dryer operated at an inlet temperature of from 110 to 120 ° C, at a flow rate of 5 ml / min, and at a temperature at the outlet of 70 to 80 ° C, which allows to obtain a fine white amorphous powder.

The powders contained the following hygroscopic growth inhibitors (HC1): ERRS. cyclodextrin, hydroxyethyl starch, dextran, dextranomer, maltodextran, hydroxypropylcellulose, hydroxypropylmethylcellulose, as well as ethylhydroxyethyl ether ethercellulose.

Powders that do not contain HC1 had the following composition (by weight): 5% salmon calcitonin, 6.25% H8A, 73.75% mannitol, 15% citrate. Powders that contain HC1 had the same relative amounts of salmon calcitonin, H8A, mannitol and citrate, but also from about 10 to 90% by weight of hygroscopic growth inhibitor.

The obtained powders are stored in tightly closed containers in a dry environment (<10% EH) until the analysis of hygroscopic growth of powders.

B. Analysis of powders

The particle size distribution for calcitonin salmon powder (KSacNopsch) was found using liquid centrifugal sedimentation carried out in a Hopa CAPA-700 type particle size distribution analyzer, followed by dispersing the powders using a 8eL18reg8e A-11 device (M1gosch1c1C5C5C5), using a 8eL18reg8e A-11 device (M1Schosch1c1C1C5C5). SA).

The moisture content of calcitonin salmon powder was measured according to Karl Fischer’s method using a hygrometer of the type MIKI CA06 Mo181igé Meter.

The distribution of aerosol particles in size was found using a cascade hammer crusher (Sgauber Aplagkep company, 8 shuta, SA) at a flow rate of 28 l / min, ignoring the loss of powder in the intake manifold and on the zero-stage jet plate.

Evaluation of released doses (ΕΌ) was carried out using an aerosol device of Therapeutic Inhalation Systems, such as described in XVO 96/09085. The released dose was determined as a percentage of the nominal dose contained in the blister pack that comes out of the mouthpiece of the aerosol device and is captured on a glass fiber filter (Ce1tap type, diameter 47 mm) to which a vacuum is applied (30 l / min) for 2.5 s after operation of the device. ΕΌ is calculated by dividing the mass of the powder collected on the filter by the mass of powder in a blister pack.

The ΕΌ value for 5% of salmon calcitonin powders is from 52.6 to 53.9%.

The value of MMAE for 5% salmon calcitonin powders is approximately from

3.5 to 3.6 microns.

The same analyzes were carried out for salmon calcitonin powders containing ORRS.

B. particle growth

The following study was conducted to study the bioaccumulation of salmon calcitonin formulations entering the lungs as dry powder aerosols.

Calcitonin salmon powders containing no hygroscopic growth inhibitor were administered to 16 healthy volunteers, each of whom received a dose of dry powder aerosols. This dose of aerosol (approximately

2.5 mg of powder) contained approximately 750 ΐυ (125 µg) of salmon calcitonin, radioactively labeled with 10 MVS. | ^99t TC pertechnitat. The creation of aerosols of particles was carried out using the above-described aerosol device of Therapeutic Inhalation Systems.

The dose of calcitonin salmon that entered the lungs and the periphery of the lungs (deep lungs) was determined using a modification of the standard gamma camera. Calcitonin of salmon entering the systemic circulation was quantified using isotope immunological analysis of serum samples taken within 6 hours after receiving the dose (using a supersensitive kit for quantitative isotopic immunological analysis and quantitative determination of salmon and calcitonin in serum and plasma (11 and 8h5c511 | ua raLyupptipoakkau ky horn 1Le OnaSubuye 1set of Bauga Cuque 1ps.)

Bioaccumulation was found when comparing estimates of the adjusted dose of A & C (area under the curve) with the peripheral (alveolar) dose. To determine the A & C (area under the curve), simple summation (integration) of the trapezium was used. Relative bioavailability was determined by comparison with subcutaneous injection. Statistical analysis of the data for the deposition in the gamma chamber and data for relative bioaccumulation was performed using the sign ranks method with selected Wilcoxon pairs (Psochopalislaus-P1H8Capeslacccc 1ec1). The Wilcoxon test is a non parametric test that is suitable for a small sample size. A p value <0.05 was considered significant.

The particle size distribution of the powder, which does not contain HC1, mainly remains unchanged before and after the radioactive labeling process. When analyzing the pattern of powder distribution by regions after inhalation, it was found that 21.6% of the dose of powder after inhalation reaches the periphery of the lungs (and the entire lung volume, taking into account the periphery of the lungs, reaches 45.6% of the powder dose), while mouth and mouth of the pharynx make up 54.4%.

The bioaccumulation of calcitonin salmon powder compared to subcutaneous injection is 28.0% for the dose entering the peripheral (deep) lungs. Although a MMAE value of 3.5 was obtained for the size distribution of aerosols, this powder behaves similarly to a powder having a MMAE from 5 to 6 microns. This can be attributed to the hygroscopic growth of particles as they pass through the airways, in accordance with the hygroscopic nature of the composition.

The justification for such a mechanism can be obtained by calculating the ratio of the growth of aerodynamic equilibrium for the composition, which is equal to 2.53. This ratio shows that, while in the respiratory tract, particles grow, at equilibrium, from an initial MMAE of 3.5 μm to a MMAE of 9 for aerosols (i.e., the particle diameter increases 2.53 times initial aerodynamic diameter). The equilibrium growth ratios are determined by calculating the concentration of solids (powder-water ratio) at which the aqueous solution of the powder becomes isotonic, that is, at a concentration at which a drop of liquid reaches equilibrium in the lungs; This then allows the calculation of ΜΜΆΌ for an isotonic drop. The growth ratio is defined as the quotient of ΜΜΆΌ for an isotonic drop by ΜΜΆΌ of the powder particles at ambient conditions.

In accordance with the present invention, to prepare salmon calcitonin powders, which retain a ΜΜΆΌ value of 3.0 μm when these powders enter the alveoli, one or more hygroscopic growth inhibitors are introduced into the particles in concentrations ranging from about 10 to 90%, and in particular, 10, 20, 30, 40, 50, 30, 60, 70, 80 and 90% by weight. The resulting powders are subjected to tests as previously described here to determine their hygroscopic growth, if any, when exposed to the medium present in the lungs. The powders in accordance with the present invention do not have hygroscopic growth or have reduced hygroscopic growth, and more specifically, support the particle size distribution with less than 3.0 μm when entering the peripheral lungs, resulting in increased powder deposition and increased bioavailability in them. lung active ingredient coming through the respiratory tract.

Example 2. Measurement of powder characteristics in an artificial climate chamber.

Obtained after spray drying powders that contain one or more inhibitors of hygroscopic growth, were prepared in accordance with example 1. The relative weight proportions of the components of the powder are given in table. one.

To assess the hygroscopic behavior of aerosol powders, dry powders intended for inhalation are placed in an artificial climate chamber in which they imitate the physiological conditions in the human lung (temperature 32 ° C and relative humidity (ΚΗ) 95%). The current control of the camera is carried out using pre-calibrated probes for measuring humidity and temperature, which allows you to constantly maintain a temperature of 32 ° C and a relative humidity of 95% in the chamber for a long period of time.

The released dose and particle size are measured under standard conditions (24 ° C and 40%), and in humid conditions (32 ° C and> 95%). Measurements under standard conditions are carried out inside the chamber with the control system turned off. The data obtained under standard conditions are used as control baseline data. Membrane filters (47 mm) are used to collect the released doses, and Andersen's cascade hammer crusher is used to measure particle size distribution.

Table 1

Sample No. The consignment No Composition ΕΏ, n. = 28 Oak. Wednesday ΕΏ Camera Sorption rate one K.98403 60% maltodextrin 2.6% glycine 4.3% mannitol 13.09% citrate 0,013 citric acid, pH 76.94 67,12 5.3 2 K98403 same 78.51 65.31 5.3 3 K984 14 40% ΙΙΕ3-Ιιιιιι \ ν 20% insulin 2.6% glycine 10% mannitol 27.4% citrate pH7.3 72.63 47.52 6.4 four K98414 same 77.23 49.39 6.4 five K99041 60% ΙΙΕ3-Ιιιιιι \ ν 20% insulin 2.6% glycine 4.3% mannitol 13.09% citrate 0.013% citric acid pH7.3 75.83 54.55 6 K99041 same 74.23 55.48

* ΗΕ8-1ιππν = gndroxyethyl starch

Given in Table. 1, the results show that содержащие-containing powders are highly dispersed under ambient conditions and retain good dispersion when simulating conditions in the lungs. These results also show how the compositions can be optimized when the amount of изменении changes, which can be seen for samples 3/4 in comparison with samples 5/6, when increasing the percentage of hydroxyethyl starch from 40 to 60% effectively reduces the drop in high humidity conditions (BP _atb1ep1 - ΕΟ | _ιιιιμ ). This indicates the resistance of содержащих-containing powders to water absorption and their ability to maintain flowability, even under conditions of very high humidity.

The value ΜΜΑΌ for powder K98403 (samples 1 and 2 in Table 1) were found under conditions of moderate temperature and humidity (70 ° E (21 ° C) and 40% relative humidity). The results obtained are shown in the table below.

Filling weight, mg (dose) ΜΜΑΏ, mkm % <5 microns % <3.3 microns five 2.9 84 59 five 2.9 84 59 2 2.3 96 77 2 2.4 94 74

From the above results it can be concluded that the specified powder retains a low ΜΜΑΌ value, even under conditions of high temperature and humidity.

Example 3

Dry powders are prepared as described in example 1. Moisture sorption profiles for powders obtained by spray drying and containing HC1. determined using a gravimetric sorption analyzer, such as an analyzer of the type δνδ-1, manufactured by νΤΙ SogrogDoi (H1ailei (Hialea), Finland). The powders obtained by spray drying have the following composition (weight percent):

Α. 20% insulin, 59% sodium citrate, 18% mannitol, 2.6% glycine

B. 100% Dextran

C. 100% hydroxypropyl methylcellulose

Ό. 100% hydroxypropyl-in-cyclodextrin

E. 100% hydroxyethyl starch (low molecular weight, ΜΑ = 200,000)

E. 20% insulin, 40% hydroxyethyl starch, 2.6% glycine, 18% mannitol, 19% sodium citrate

6. 20% insulin, 20% leucine, 50% β cyclodextrin sulfonyl butyl ether, 10% sodium citrate

H. 20% insulin, 20% leucine, 50% hydroxyethyl starch, 10% sodium citrate

Ι. 20% insulin, 5% leucine, 50% hydroxyethyl starch, 25% sodium citrate

I. 20% leucine, 50% hydroxyethyl starch, 30% sodium citrate

Moisture sorption profiles for powders A, B, C, Ό and E are shown as graphs in FIG. 2

Moisture sorption profiles for powders, E, and E are shown as graphs in FIG. 2

Moisture sorption profiles for Α, Ό, and C powders are shown as graphs in FIG. four.

Moisture sorption profiles for powders H, I, E, A and 1 are shown as graphs in FIG. five.

From consideration of each of the graphs, we can conclude that the addition of hygroscopic growth inhibitor to the powder effectively and significantly reduces its moisture sorption properties, resulting in reduced hygroscopic growth of particles as they move along the respiratory tract into the deep lungs.

Example 4

Comparison of the properties of the hydration of various spray-dried insulin powder formulations obtained was performed by monitoring the thermal activity of TAM using the thermal activity monitor TAM, άοе1 2277 (Täögtotör1s, Sweden). Test conditions were as follows: linear change mode using HV perfusion units; a change from 0% HV to 90% HH with a change of 3% HH per hour at 25 ° C, with a flow of nitrogen 1.48 8ΟΟΜ. The following powder formulations were used:

Α. 20% insulin, 59% sodium citrate, 18% mannitol, 2.6% glycine

B. 60% (wt.%) Insulin, 2.6% glycine, 10% mannitol, 27.4% sodium citrate, 2.6% glycine.

C. 40% HE8-ytet, 20% insulin, 2.6% glycine, 10% mannitol, 27.4% sodium citrate

Ό. 60% maltodextrin, 20% insulin, 2.6% glycine, 4.3% mannitol, 13.09% sodium citrate, 0.13% citric acid.

The results of the TAM analysis for each of the compositions are shown in FIG. 3. From consideration of FIG. 3 we can conclude that the introduction of HC1 in formulations with 20% insulin leads to a significant drop in both the rate and degree of moisture absorption compared to the composition with 20% insulin that does not contain HC1, which indicates the effectiveness of hygroscopic growth inhibitors to reduce the degree of hygroscopic growth of such particles.

Claims (24)

CLAIM 1. A powder for delivering the active substance to the patient’s alveoli, containing the active substance and a hygroscopic growth inhibitor, characterized in that the hygroscopic growth inhibitor is introduced into the particle volume in an amount of at least 40 wt.% And is selected from the group consisting of β-cyclodextrin, hydroxypropyl-b-cyclodextrin, sulfobutyl ether-b-cyclodextrin, dextranomer, hydroxyethyl starch and maltodextrin, while the particles have a drop in dose when the lung conditions are simulated, not exceeding approximately 25%. 2. The powder according to claim 1, characterized in that the hygroscopic growth inhibitor is selected from the group consisting of hydroxyethyl starch, dextranomer and maltodextrin. 3. The powder according to claim 1, characterized in that a cyclodextrin selected from the group consisting of β-cyclodextrin, hydroxypropyl-b-cyclodextrin, sulfobutyl ether β-cyclodextrin is used as an inhibitor of hygroscopic growth. 4. The powder according to claim 1, characterized in that hydroxyethyl starch is used as an inhibitor of hygroscopic growth. 5. The powder according to one of claims 1 to 4, characterized in that the hygroscopic growth inhibitor is present in the powder in an amount sufficient for the powder to have a moisture absorption rate not exceeding approximately 0.50% as a function of relative humidity. 6. The powder according to one of claims 1 to 5, characterized in that the hygroscopic growth inhibitor is present in the powder in an amount sufficient for the powder to have a total moisture absorption not exceeding approximately 30% by weight at a relative humidity of 80%. 7. The powder according to one of claims 1 to 6, characterized in that its released dose in environmental conditions is at least 60%. 8. The powder according to one of claims 1 to 7, characterized in that the powder particles contain approximately from 40 to 99 wt.% The hygroscopic growth inhibitor. 9. The powder according to one of claims 1 to 8, characterized in that upon delivery to the lungs, the deposition of the powder in the deep lungs exceeds 20% of the nominal dose. 10. A powder according to one of claims 1 to 9, characterized in that the hygroscopic growth inhibitor effectively increases the bioavailability of the active substance when delivered to the lungs by at least 5% compared with the bioavailability of the active substance when delivered to the lungs for particles, not containing an inhibitor of hygroscopic growth. 11. A powder for delivering the active substance to the patient’s alveoli, containing the active substance and a hygroscopic growth inhibitor, characterized in that the content of the hygroscopic growth inhibitor is at least 40 wt.%, While the particles have a sorption rate of approximately less than 6.5, and the inhibitor is selected from the group consisting of β-cyclodextrin, hydroxypropyl-b-cyclodextrin, sulfobutyl ether-β-cyclodextrin, dextranomer, hydroxyethyl starch and maltodextrin. 12. The powder according to claim 11, characterized in that the hygroscopic growth inhibitor is selected from the group consisting of hydroxyethyl starch, dextranomer and maltodextrin. 13. The powder according to claim 11, characterized in that hydroxyethyl starch is used as an inhibitor of hygroscopic growth. 14. The powder according to one of paragraphs.11-13, characterized in that its released dose in environmental conditions is at least 60%. 15. A powder according to one of claims 11-14, characterized in that it contains from about 40 to 99% by weight of an inhibitor of hygroscopic growth. 16. A powder for delivering the active substance to the patient’s alveoli, containing the active substance and a hygroscopic growth inhibitor, characterized in that the hygroscopic growth inhibitor is introduced into the particle volume in an amount of at least 40 wt.%, Selected from the group consisting of β-cyclodextrin , hydroxypropyl-b-cyclodextrin, sulfobutyl ether-b-cyclodextrin, dextranomer, hydroxyethyl starch and maltodextrin, while the powder maintains an aerosol particle size distribution profile below 3 μm ΜΜΆΌ upon delivery to the alveoli m. 17. A method of preparing a powder for delivery of the active substance to the patient’s alveoli, characterized in that it includes the following operations: preparation of a mixture of a hygroscopic growth inhibitor selected from the group consisting of β-cyclodextrin, hydroxypropyl-β-cyclodextrin, sulfobutylether-b-cyclodextrin, dextranomer, hydroxyethyl starch and maltodextrin, the active substance and the solvent, while the amount of inhibitor is at least 40% of total weight of the powder; and spray drying the mixture to obtain homogeneous particles of an inhibitor of hygroscopic growth and the active substance, and the particles have a drop in the released dose when simulating conditions in the lungs, not exceeding approximately 25%. 18. The method according to 17, characterized in that the hygroscopic growth inhibitor is selected from the group consisting of β-cyclodextrin, hydroxypropyl-b-cyclodextrin, sulfobutyl ether β-cyclodextrin. 19. The method according to 17, characterized in that the hydroxyethyl starch is used as an inhibitor of hygroscopic growth. 20. The method according to one of paragraphs.17-19, characterized in that the solvent is water. 21. The method according to one of paragraphs.17-20, characterized in that the homogeneous particles contain from about 40 to 99 wt.% The hygroscopic growth inhibitor. 22. A powder made by the method according to 17. 23. The powder according to any one of claims 1 to 16 in aerosol form, intended for delivery of the active substance to the lungs of the patient. 24. The powder according to any one of claims 1 to 16, designed to deliver the active substance to the patient’s lungs using a dry powder.