Dried spray particles as therapeutic carriers


(57) Abstract:

The invention relates to medicine. Microparticles of water-soluble material, which are smooth and spherical, and at least 90% of which have a mass-average particle size of 1 µm and 10 µm, which are therapeutic or diagnostic agent, or use of this agent as a water-soluble material can be successfully used in inhalers, spray dry powder for delivery of the specified agent. The proposed particles provide more effective delivery of drugs and diagnostics in the alveoli. 5 C. and 14 C.p. f-crystals, 11 PL.

The invention relates to a dried dispersion of the microparticles and their use as therapeutic carriers. More specifically, the present invention relates to the delivery of diagnostic and therapeutic agents and products of biotechnology, including medicines obtained using rDNA technology.

The most common route of administration of therapeutic agents, oral or gastrointestinal largely not applicable to peptides and proteins derived using rDNA technology. The sensitivity of normal peptide is edenia. Logical means of injection is intravenous infusion, but it presents certain difficulties associated with poor patient compliance with the regimen and the regimen with a long introduction and, very often, with rapid clearance after the first pass through the liver, resulting in a short period of/in life.

Recently studied the possibility of delivery by migrating through the mucous membranes. At that time, as a nasal delivery has been extensively studied, it is possible to deliver peptides via the pulmonary route remains largely unexplored.

Alveolar cells are inherently effective barrier. However, even conducting material in the area of the alveoli is a significant obstacle to this method of introduction. There is an optimal particle size that allows them to achieve the lower divisions pulmonary tract, i.e., aerodynamic diameter < 5 μm. Particles, larger particles remain in the upper part of the respiratory tract so that the standard commercial suspension drugs only 10 - 30% of the particles, normally polydispersion suspension, reach the lower respiratory tract.

Modern methods Aero is asplenia aqueous solutions requires large amounts of drugs and bulky and non-handheld devices.

The most common route of administration of drugs to the lungs is the use of devices with volatile propellant, commonly referred to as a metering device. Their main structural characteristic is the solution of the propellant, usually CFC 11, 12 or 114 containing dissolved drug or suspension of the drug in the container under pressure. The dosing is done by pressing on the actuating mechanism, which releases propellents aerosol drug suspension or solution, which is delivered to the respiratory tract. During passage through the lungs, the propellant evaporates, forming microscopic precipitation from solution or loose particles from the suspension. This dosage is easily reproducible and cheap, but the growing environmental pollution forced to reduce the use of CFC. Moreover, the use of CFC solvents remains largely incompatible with many modern drugs are derived by means of biotechnology, because of their susceptibility to denaturation and low stability.

At the same time, there is a tendency to use the device with a dry powder, which use dry powders of drugs, obscestva. Energy disaggregation often provides breathing or inhalation of air through the device.

Currently, medications are milled to reduce particle size. This approach is not applicable for products derived by biotechnology. In General, the products obtained by means of biotechnology, available in small quantities and, moreover, they are sensitive to the methods currently used for drying and grinding, prior to mixing with the filler. Further, it is especially difficult to produce a mixture of drug and excipient is fluid enough so that they flowed and was desirables in a reproducible manner in the modern mnogochasovykh inhalers, such as Turbohaler (Astra) and Diskhaler (Glaxo). Studies have shown that contrary to expectations, spray dried (spherical) particles of salbutamol, possessed by the forces of cohesion and adhesion greater than the particles of powdered drugs of the same size. Pictures spray dried material obtained using an electron microscope showed that the particles have jagged, rough surface.

Haghpanah et al. in 1994 the British pharmaceutical conference reported receiving dried races is piratory drug delivery, i.e., 1 to 5 μm. The goal was encapsulating salbutamol for slow its release. This becomes evident that the product consists of a substantially homogeneous spherical and smooth particles having the characteristics of fluidity, satisfactory for use in mnogochasovykh inhalers dry powder.

Diagnostic agents, including hollow microcapsules, are used to enhance the image with an ultrasound. For example, EP-A-458745 (Sintetica) discloses a method of manufacturing a filled with air or gas microballoon by polymerization on the surface of the partition synthetic polymers, such as polylactide and polyglycolides. WO-A-9112823 (Delta) discloses a similar method using albumin. Wheatley et al. (1990) Biomaterials 11: 713-717 reveal ionotropic the gelatinization of alginate to obtain a microbubble diameter is more than 30 μm. WO-A-9109629 discloses liposomes for use as contrast agents for ultrasound.

Przyborwski et al., Eur. J. Nucl. Med. 7:71-72 (1982) disclose the preparation of microspheres of albumin human (CSA) using spray drying for the purposes of the radioactive treatment and their subsequent Primaire this work was mostly poorly designed solid microspheres. If the particles are not hollow, they are not suitable for echocardiography.

In addition, these microspheres were made using a one-step process, which, as we have established, is not suitable for the manufacture of microcapsules suitable for echocardiography; in this process it was necessary to remove from the microspheres Undenatured albumin, and was clearly out of the microspheres, varied greatly in size, requiring the additional step of screening.

Przyborowski et al. refer to two earlier descriptions of methods for producing particles of albumin for lung scintigraphy. Aldrich and Johnston (1974), Int. J. Appl. Rad. Isot., 25:15-18 disclose the use of a rapidly rotating disk to obtain particles with a diameter of 3 to 70 μm, which are then denatured in hot oil. The oil is removed, and the particles have been labelled with radioactive isotopes. Raju et al., (1978), Isotopenpraxis 14(2):57-61 used the same technology with the rapidly rotating disk, but albumin was denaturiruet simple heating of the particles. In none of these cases was not mentioned hollow microcapsules, and produced particles were not convenient for echocardiography.

EP-A-0606486 (Teijin) describes the production of powders, in which the active agent is incorporated into small particles, with socialearth to gelatin capsules, used in the dosing inhaler dry powder. Page 12 of this publication mentions the spray drying "drug basis" to obtain particles of which 80% or more have a size of 0.5 - 10 μm. There is no guidance as to what conditions should be used to obtain such a product.

EP-A-0611567 (Teijin), more specifically, relates to the production of powders for inhalation by spray drying. The carrier is a cellulose selected because of its resistance to humidity environment. The terms and conditions mentioned in example 1 (ethanol as solvent, 2 - 5% (weight/volume) of a dissolved substance) indicate that the surface structure is not regulated, and example 4 reports the fraction of poorly penetrating into the lower respiratory tract by inhalation (12%), which indicates poor dispersion properties. Spherical particles receive clearly the high content of drugs, which indicates the dependence structure of the particles from the respective content medications and media.

Conte et al. (1994), Eur. J. Pharm. Biopharm. 40(4): 203 - 208 describe the spray drying of the aqueous solution with a minimum content of dissolved substances of 1.5%. To get the majority of the particles are almost spherical shape, requires a high content of the e suspension in butanol, to facilitate analysis Coulter is required sonication, implying that the particles are not completely dry.

The aim of the present invention is to provide a carrier for drug delivery and compositions, which are better suited than previously known products for delivery, in particular in the alveoli.

The invention

According to the present invention it has been unexpectedly discovered that the microparticles and microcapsules and microspheres), which is also convenient as an intermediate product, i.e., before fixation, in case of production containing air microcapsules for diagnostic purposes, for example, described in WO-A-9218164 as "intermediate microcapsules", forming the wall material spray drying has virtually no effect. Thus, it is possible to produce and present in the form of dry powders for therapeutic and diagnostic use in a high degree homogeneous microparticles, microspheres or microcapsules of heat-sensitive materials such as enzymes, peptides and proteins, for example, CSA, and other polymers.

In contrast to the previously known state of this region, currently framewith by spray drying, which are smooth spherical microparticles with good fluidity of the water-soluble material, such as serum albumin human (CSA) having a mass-average particle size of from 1 to 10 μm. In a more General sense, the process of production of microcapsules of the present invention involves the atomization of a solution (or dispersion) of the material forming the wall. Drug or diagnostic agent can be atomserver immediately or attached to the microcapsules obtained in this way. Alternatively, the material itself may represent an active agent. In particular, it was found that under the conditions given in this document and in more General form described by Sutton et al. (1992), for example using an appropriate combination of higher solute concentrations and higher ratios of flow of the air/liquid than Haghpanah et al., and amplifiers education sheath, it is possible to produce extremely smooth spherical microparticles of different materials. The spherical nature of the particles can be installed not only simple definition of the maximum size, i.e., using the method of diffraction of the laser beam described Haghpanah et al. Moreover, the size of Catinaccio. For example, analysis Coulter, 98% of the particles can be less than 6 μm on a numerical basis, within the limits of the interquartile range 2 μm, and with an average size of variations between the parties is less than 0.5 μm. Further, when testing in the inhaler dry powder on the stage of development was reached reproducible dosing and subsequent aerosolization under normal conditions of flow (30 l/min) showed excellent separation of particles from the filler.

Flow of capsules of the present invention obtained from Undenatured CSA or other material able to be dried by spraying, have a very smooth surface and can be produced with a relatively low filler content to obtain wysokosci powders, ideal for use in inhalers dry powder. Using this approach, you can produce heterogeneous microcapsules consisting of suspended fillers and active ingredient. This method has the advantage that you can get wysokosci powder active ingredients that can be subjected to further processing to produce powders that are proportioned to form an aerosol with excellent reproducibility and accuracy.

the rotation of the polymers in the production of vysokoletuchih powder. In all cases, the size of the suspension of microcapsules may be such that 90% of the mass was within the desired size, for example size suitable for inhalation, 1-5 microns.

So, essentially we have described how to produce microparticles that have a size predominantly 1-5 microns, are smooth and spherical, contain gas and consist of intact protein molecules and which can be stored and transported to other stages of production. For the manufacture of intermediate microcapsules for ultrasound we brought those characteristics of the process and the resulting powder, which are essential for the production of excellent powders for inhalers, spray dry powder (COI). We found that many of the analyses that are designed to echocontrast agents useful for determining the parameters of the particles, which are useful for use in COI powders, namely echo and pressure resistance of cross-linked particles that define a well-made microparticles; microscopic evaluation in DPX or solvents determining the sphericity and the contents of the eyes in a soluble intermediate capsules, an level of fixation of the product.

Considerable attention should be paid to control of particle size and size distribution, especially in products that are used in therapy. We chose a biologically compatible polymer which when cross-linking remains harmless, and learned how to reproducibly perform the cross-linking of this molecule. In order to achieve the controlled binding, we have divided the formation of microparticles and cross-linking that was not done in other processes such as evaporation of the emulsion and solvent. This means that the initial stage of the process does not damage the material forming the wall. We have identified the specific parameters relevant to the complete formation of the particles, and also identified more favorable conditions under which the output of intact particles is much better. Selects CSA as a particularly suitable polymer, we also chose a potential molecule-carrier, which can protect labile molecules, to enhance the absorption of peptides in the lungs, to bind low molecular weight drugs due to the natural affinity and covalently modified to carry drugs across cell barriers in the system, the circus, sledovatel were prone to use of volatile solvents, which cause rapid shrinkage of the droplets. Alternatively, the researchers used a raw material solution having a low content of dissolved substances to maintain its low viscosity to facilitate the acquisition of droplets of a smaller size. In both cases, the production of microparticles method has little effect on the final structure; rather, it is dictated by the components that are used for the formation of particles. We undertook extensive research regarding how it is possible to make particles from CSA with controlled size, and have applied it to many other materials, including active drugs. We can use a relatively high content of dissolved substances, for example, 10-30% (weight/volume), as opposed to 0.5-2%, for the manufacture of microparticles comprising a low molecular weight active agent and lactose; only low-molecular active agent; a peptide with CSA and modified polymeric carriers with the active agent. We found that the method determines the final structure of the particles to a greater extent than the composition of the dissolved substances. Next, we can use a combination of water and miscible with water Rast is manual, which gives the opportunity to produce a smooth spherical particles with controlled size, convenient for delivery into the lungs.

We found that the method of the present invention can be modified in order to obtain microspheres with the desired characteristics. Thus, the pressure at which the protein solution is fed to the nozzle of the spray can be varied, for example, in the range 1.0-10.0 105PA, preferably, 2.8105PA and, most preferably, about 7.5 105PA. Other parameters can be changed as described below. This way you can get new microspheres.

In accordance with another aspect of the present invention provided with hollow microspheres, of which more than 30%, preferably more than 40%, 50% or 60% of the microspheres have a diameter within the span of 2 microns and at least 90%, preferably at least 95%, or 99% have a diameter within the scope 1.0-8.0 μm.

Interquartile spread may be 2 μm with an average diameter 3.5, 4.0, 4.5, 5.0, 5.5, 6.0 or 6.5 μm.

Thus, at least 30%, 40%, 50% or 60% of the microspheres may have a diameter in the range of 1.5-3.5 μm, 2.0-4.0 μm, 3.0-5.0 μm, 4.0-6.0 μm, 5.0-7.0 μm or 6.0-8.0 μm. Preferably, the specified percentage km, 5.0-6.0 μm, 6.0-7.0 μm or 7.0-8.0 μm.

Another aspect of the present invention creates a hollow microspheres with protein walls, of which more than 90%, preferably at least 95%, or 99% of the microspheres have a diameter in the range 1.0-8.0 μm; at least 90%, preferably at least 95%, or 99% of the microspheres have a wall thickness of 40-500 nm, preferably, 100 to 500 nm.

Constituting the wall material and process conditions should be chosen so that the product was sufficiently non-toxic and non-immunogenic when conditions in which it will be used, which obviously depend on the input dose and duration of treatment. Forming the wall material may be derived starch, a synthetic polymer, such as tert-butyloxycarbonyl (US-A-4888398) or polysaccharide, such as Polydextrose.

In General, constituting the wall material can be selected from the most hydrophilic, biodegradable physiologically compatible polymers, as described in more detail in WO-A-9218164.

Preferably, forming the wall material has a protein nature. For example, it may be a collagen, gelatin or serum) albumin, cardable preferably, it is human serum albumin (CSA) obtained from donor blood, or, ideally, from microorganisms (including cell lines) that have been transformed or transfectional for expression CSA. Additional details are listed in WO-A-9218164.

The protein solution or dispersion has a concentration of protein, preferably from 0.1 to 50% (weight/volume), more preferably about 5.0 to 25%, especially when the protein is albumin. The value of about 20% is optimal. You can use the mixture forming the wall materials; in this case, specified in the last two sentences percentages refer to the total content constituting the wall material.

The drug is designed for spraying, can contain, in addition to constituting the wall material and liquid solvent or carrier, and other substances. Here again, you can refer to WO-A-9218164.

The protein solution or dispersion (preferably a solution), hereinafter referred to as "protein preparation", atomizer and spray dried in any convenient way, resulting in a gain of discrete microspheres or microcapsules with a diameter of from 1 to 10 μm. These figures are at least 90% of the microcapsules, dia is in, filled with gas or steam, but not any solid materials. Particle type honeycomb, reminiscent of confectionery products sold in the UK under the trade name Malteserswhile not formed.

Automating involves the formation of an aerosol of the protein preparation by, for example, the bandwidth of the drug under pressure through at least one hole, or through the use of centrifugal atomizer in the chamber with hot air or other inert gas. Luggage must be large enough for the largest of ejected droplets do not hit the wall before drying. The gas or vapor in the chamber is clean (i.e., preferably sterile and free of pyrogens) and non-toxic when introduced into the bloodstream in amounts related to the introduction into the body of the microcapsules when you use them. The rate of evaporation of liquid from the protein preparation should be high enough so that the microcapsules was hollow, but not so high that they exploded. The evaporation rate can be adjusted by adjusting the flow rate of gas, the concentration of protein in the protein preparation, the nature of the liquid carrier, the feed rate of the solution and that more VA is over when the intake, at least about 100oC, preferably at least 110oC is usually sufficient to ensure that small capsules, and may be higher, up to 250oC, without the risk of rupture. The temperature was about 180 - 240oC, preferably about 210 - 230oC and most preferably about 220oC is optimal, at least for albumin. As the temperature of the strip facing the spray, also depends on the speed of delivery of aerosol and liquid content in the protein preparation, the temperature at release can be monitored in order to ensure adequate temperature in the chamber. It was found that a suitable temperature when the release is 40 - 150oC. it Was also found that the regulation of flow rate suitable to control other parameters such as the number of intact hollow particles.

Microcapsules typically include 96 - 98% Monomeric, CSA.

More specifically, the microparticles of the present invention preferably have a maximum interquartile span of 3 μm, more preferably, 2 μm, and most preferably, 1.5 μm, according to their mass-average particle size. The mass-average diameter, the I by spray drying, where there is a combination of low flow rate of the raw solution with high levels of automation and drying air. The effect is to develop microcapsules very specific size and narrow size distribution.

Several employees brought equation to determine the average size of the cells when using pneumatic nozzles; a simple version of the various parameters that affect the average size of the droplets, as follows:

D = A/(V2d)a+ B (Mair/Mliquid)-b,

where D is the average size of the droplets;

A - constant associated with the design of the nozzle;

B is a constant related to the viscosity of the liquid;

V is the relative velocity between the fluid and the nozzle;

d is the density of air;

Mairand Mliquid= mass flow of air and liquids;

a and b are constants associated with the design of the nozzle.

Obviously, when any design nozzles on the size of the droplets are most affected by the relative velocity at the nozzle and at the same time the mass ratio of air and fluid. For the most common applications with the purpose of drying, the ratio of air to fluid nah the effect of microparticles in the size range, specified in this document, we used the ratio of air and fluid in the range of 20 to 1000 : 1. The effect is the production of particles at high ratios, which are very small in comparative standards, with very narrow size distributions. For microparticles produced at lower proportions of air and fluid, their size is slightly bigger, but they still, however, have a narrow size distribution that exceed those of the microparticles produced by emulsion technologies.

The amount added of the active component is not important; microparticles can include at least 50, more preferably 70 or 80, and most preferably, 90 wt.% CSA or media of a different material. For use in the inhaler microparticles can be placed in a composition with a conventional filler, such as lactose or glucose.

Microparticles can include a therapeutic agent and a carrier or connection, which in itself is therapeutically active. The number of the active component should be selected, taking into account its nature and activity, the route of administration and other factors known Spa is 100 mg/day l or 0.1 g/day of active material, such as beclomethasone.

The active ingredient may comprise, for example, diagnostic classic pharmaceutical substance or object that can be linked or not linked, covalently or otherwise, with the material carrier. therapeutic agent can comprise a protein material such as insulin, parathyroid hormone, calcitonin or similar biologically active peptide, albuterol, salicylate, naproxen, Augmentin or cytotoxic agent. For experimental purposes may be included marker, such as lysine-fluorescein.

Microparticles of the present invention may include, in addition to therapeutic or diagnostic agent, antagonist or binding the receptor component. For example, in molecular media, you can include sugar or another molecule, referring to the directed introduction associated with a carrier of the drug to the receptor on the alveoli or later.

CSA in this document is used as an illustrating example of water-soluble materials of nositelei for use in the present invention. Other materials that can be used include simple and complex carbohydrates, simple and enantia human proteins or their fragments or short form.

The present invention allows to manipulate the nature of the dry microcapsules to optimize the flow properties or media by changing and reducing the forces of cohesion and adhesion within the drug microparticles. For example, if you need, it is possible to produce microcapsules, which are in advance will bear a positive or negative charge, by using vysokonapryazhennyh Monomeric or polymeric materials, such as lysine, or polylysine and glutamine, or polyglutamate systems without CSA, or heterogeneous systems, including CSA and active components.

Another embodiment of the present invention is the drying of the joint dispersion of the active component and CSA aimed at facilitating the stabilization of the active component during manufacture of the composition, packaging and, more importantly, during your stay on the alveolar epithelium. In this environment may be high proteolytic activity. While for the protection of medicines peptide can be used protease inhibitors, may exist and contraindications to this approach. Using CSA, both as a filler and as a carrier, you can try a large number is is then, because CSA, as shown, passes through the alveolar barrier, through translatory mechanisms, mediated or not mediated by receptors, it can be used as a carrier to facilitate the passage of the active ingredient through the epithelial lining.

In yet another embodiment, the active component before spray drying can be covalently linked to CSA through split links. This alternative implementation is the method of transport of active ingredients all the way from the device to the blood flow and possibly to the targets within the body. The formation of particles with optimal aerodynamic size, means that the "physical" media delivers the active ingredient to the site, which is its absorption. Otlozhilis on the alveoli, the "molecular" media then protects the active component and facilitates its origin in the bloodstream and, once in the bloodstream, could further increase the half-life in circulation, and even to direct the active ingredient to specific areas inside the body using mediated by receptors phenomena.

Convenient technology linking agents described what I technology, applied for the derivation of CSA before spray drying, makes possible the production of covalent system carrier for drug delivery into the systemic vasculature. In this case, you use the ability CSA to pass through the alveoli to transfer drugs for a long period of time, at the same time protecting potentially unstable objects.

Despite the fact that the active component used in the present invention, can be impregnated microparticles or any other way to attach it to them after their manufacture, it is preferable to connect it in combination with CSA. Microparticles can be coated, at least partially hydrophobic or water insoluble material such as a fatty acid, to lower the rate of dissolution and to protect against swelling in the aquatic environment.

The following examples illustrate the present invention. Drying spray, which is used in the examples and which can be purchased from the company A/S Niro Atomizer, Soeborg, Denmark, under the trade name "Mobile Minor" detail in WO-A-9218164.

Example 1

20% solution of sterile pyrogen-free, CSA in pyrogen-free water (for injection) pump, described above. Speed peristaltics pump was maintained at a level of approximately 10 ml/min, so that when the temperature of air at inlet 220oC, the temperature at release was kept at 95oC.

Compressed air is supplied to a two-stream spray nozzle under a pressure of 2.0 - 6.0 bar (2,0 - 6,0 105PA). When this mode is obtained microcapsules with an average size of 4.25 to 6.2 microns.

In a typical case, the increase in the average particle size (by lowering the pressure atomization) leads to an increase in the number of microcapsules with a size exceeding 10 μm (see table 1).

Under the above conditions, i.e. at the first stage of example 1 of WO-A-9218164, when the pressure in the nozzle 7.5 bar, we have obtained microparticles with a size of 4.7 μm. These soluble microparticles were smooth and spherical when the content of particles larger than 6 microns, less than 1%. These microparticles were dissolved in water and determined the molecular weight of CSA by gel chromatography. The obtained chromatogram for CSA before and after drying, CSA spray was almost the same. Additional analysis of CSA before and after drying by atomization through tripticase peptide of Cartaromana conditions of spray drying, described to obtain microparticles with a size of 4.7 μm, the structure of the protein with little or no applied damage.

Example 2

Alpha-1 antitripsin, isolated from human serum, spray dried under the condition similar to the conditions of example 1, with the intake temperature 150oC and temperature release 80oC. Other drying conditions were the same as in example 1. Made soluble microparticles had an average size of 4.5 μm. These microparticles were dissolved in water and analyzed for retention of protein structure and normal trypsin inhibitory activity, and then compared with the original liofilizirovannam source material. Analysis by gel chromatography, chromatography with reversed phase and capillary electrophoresis showed that after drying the dispersion was not observed significant structural changes. Analysis of inhibitory activity (table 2) showed that within the experimental error was achieved by fully preserving inhibitory activity.

Example 3

Using the General method of example 1 were fabricated microcapsules consisting of alcohol dehydrogenase (ADH) and lactose (ADH 0.1 wt.%, lactose to 99.9 weight. %). We have found that d is subject to the General conditions of example 1, but I changed the temperature at the inlet and outlet to obtain the conditions that have allowed the production of microparticles of the desired size (4 to 5 μm), fully maintain activity after drying and re-dissolving in the water environment. The percentage of retained activity in comparison with the original material for each of the conditions of spray drying are shown in table 3. Microcapsules were smooth and spherical and contained air, as evidenced by their appearance in diphenylsilane (DFC) under light microscopy.

Example 4

To study the effect of feed rate of the liquid raw material at the exit of intact spherical particles was performed a series of experiments under the conditions described in example 1. We found that using the ability of gas-bearing microparticles reflect ultrasound, it is possible to determine the optimal conditions to maximize the yield of intact smooth spherical microcapsules. Microcapsules obtained after spray drying, fixed by heating, to make them insoluble, and then suspended in water to measure the echo signal. We found that the increase in the rate of flow of the liquid raw material reduces the number of intact microparticles, original: open the walls, not changed, but changed the overall echogenicity increases the rate of fluid flow from 4 to 16 ml/min, We found that a slower evaporation (at higher velocities of fluid flow) leads to a decrease in the amount of intact spherical particles.

This study was conducted by resuspendable fixed by heating the particles at a concentration of 1 to 106ml 350 ml of water. This solution was slowly stirred in a 500 ml beaker, which was installed ultrasonic probe 3.5 MHz, connected to a medical device for producing ultrasonic images Sonus 1000. Received ultrasonic image capture analyzer and compared with water, which served as a control to obtain units of videolatest echo signal. This study can also be adapted to study resistance to pressure, by estimating the echo signal before and after exposure to the samples cyclical upswings pressure applied to the stock solution of particles. This analysis distinguishes incomplete particles, which after re-dissolution release the air from the fully spherical particles, which "encapsulate" the air inside the shell. Incomplete particles nya fixed albumen particles of example 1 is approximately, 5, 9, 13, 20, 22 and 24 UNITS of videolatest (intensity backscatter) at concentrations of microcapsules, respectively, of 0.25, 0.5 to 1, 2, 3, and 4 of 106on Jr.

Example 5

To reduce the particle size and narrow distribution in different sizes was conducted an important experiment. This experiment served to effectively increase the gas content in exocontact agent and reduce the number of particles greater than the desired size. This experience is also useful for creating compositions intended for administration to the respiratory tract because it maximizes the potential number of suitable for inhalation of particles in the size range of 1 - 5 µg and independently produces more smooth particles that are less cohesive in comparison with non-spherical particles of the same size.

We found that the possible reduction of particle size by reducing the content of dissolved substances in the raw material solution. This effect was partially mediated the effect of viscosity on the formation of droplets. However, we also found that the reduction of dissolved substances under the same conditions that we used, leads to a significant decrease in the number entactiniidae volatile solvent increases the rate of formation of membranes during drying with a concomitant increase in the number of intact particles or hollow particles (table 5). Evaluation of hollow microcapsules produced using the definition under the microscope the number of particles that pop up to the surface of the cover glass in hemocytometer, and compare it with the number of particles, determined by Coulter counting.

Example 6

For the manufacture of smooth spherical soluble microparticles were used a number of materials. This range includes inert materials such as CSA, lactose, mannitol, sodium alginate; active materials, such as-l-antitripsin, and a mixture of the active material and the inert carrier such as lactose/alcoholdehydrogenase, lactose/budesonide, CSA/salbutamol. In all cases were obtained smooth, spherical, containing gas particles.

We evaluated the success of the process in maintaining control over the structure of the particles. These particles are suspended in propanol, and then studied under a microscope. Particles that contain gas, have intensely white core surrounded by intact black bezel, while destroyed or badly formed particles appear as "ghosts". Microscopic

assessment the following microparticles illustrates the range of materials and active ingredients, which can be dried for the manufacture of smooth Sveta



Example 7

Lactose and budesonide spray dried under the conditions described in the table below (table 6).

The obtained dry powder was mixed with filler lactose in the mixer of type V in the proportions listed in table 7. The mixture was then placed in gelatin capsules and were unloaded from RotahalerTMin a two-stage impinger working at 60 l/min. Suitable for inhalation fraction was calculated as the percentage otlogetswe in the lower chamber.

To obtain a suitable for inhalation fractions significantly exceed the proportion of such fractions in the powdered product that is currently used in this device, which is usually located within a maximum of 10 - 20%.

Composition budesonide/lactose detailed in example 7, was tested in experimental fed by gravity mnogomodovoi COI. The studied parameters were the change in emitted dose after 30 clicks and suitable for inhalation fraction in four impingere device. The results are presented in table 8.

For modern devices COI preliminary recommendation farmacopea USA costal is avanian, in cases of compositions 1 and 2 these parameters significantly less modern restrictions.

Example 8

To reduce the dissolution rate of the soluble microcapsules, as described in the previous examples, the microcapsules may be coated with fatty acids, such as palmitic or Baganova acid. Soluble microcapsules of example 1 was coated by a suspension of a mixture of soluble CSA microcapsules and glucose (50 wt.%) in an ethanol solution containing 10% palmitic or beganovi acid. The solution is evaporated, and the resulting dense residue was washed passing through the mill Fritsch.

The effectiveness of the coating was evaluated by an indirect method, derived from our previous ultrasound studies. The ultrasound image was acquired with chemical glass of water containing 1 to 106microcapsules/ml, using an ultrasonic machine HP Sonus 1000, which is connected with the image analyzer. During the whole time measured videointernet compared with control measurements (ED videolatest) (table 9).

Microcapsules uncoated quickly lost all the air and, consequently, the ability to reflect ultrasound. However, microcapsules coated remained several minutes.

Example 9

Soluble microcapsules of mannitol were produced as described in example 1 (raw material for drying spray - 15% aqueous solution of mannitol), and covered or palmitic or beganovi acid as described in example 8. A sample of each species suspended in water and measured its echo. Ten minutes after the first analysis were measured again echo suspended samples (table 10).

Example 10

Soluble microcapsules with a model of the active compound (lysine-fluorescein) contained in the matrix, were made to make it possible to obtain active compounds in the form vysokoletuchih dry powder. Upon the dissolution of the microcapsules, the active compound was released in its native form.

When using lysine as a model compound molecule was labeled fluorescein-isothiocyanato (FITZ), so the connection could be observed during the preparation of soluble microcapsules and during its subsequent release during dissolution.

3 g of lysine was added to FITZ (total 0.5 g) in carbonate buffer. After incubation at 30oC for one hour the resulting solution was analyzed for NALIChII/lysine.

Adduct FITZ/lysine mixed with 143 ml of 25% ethanol containing 100 mg/ml CSA to produce raw materials for drying spray. The conditions of spray drying, which was used to obtain microcapsules are described in detail in table 11. We found that in the absence of ethanol is only a small fraction of particles has a smooth spherical structure.

In the process of drying by atomization was obtained 17,21 g of microcapsules, which did not dissolve when resuspending sample in ethanol. Moreover, it was not observed release adduct FITZ/lysine. However, when added to the microcapsules suspended in ethanol, 10 ml of water microcapsules were dissolved, and FITZ/lysine freed. Analysis of the adduct using thin-layer chromatography before incorporation in microcapsules and after release from the microcapsules by dissolving showed that model the connection is not changed.

Dimensions soluble microcapsules was determined in non-aqueous system of ammonium thiocyanate and propan-2-ol using a Multisizer II (Coulter Electronics). The microcapsules had an average size of 3.28 0.6 microns, and 90% of the mass was in the range of 2 to 5 microns.

Microcapsules were mixed with glucose (50% (wt./about.) microcapsules: 50% (weight. /about. ) glucose) and with predelli by TLC analysis, comparing with its original form, was recovered intact. This example demonstrates the feasibility of producing an amino acid or peptide composition that includes CSA, which can be used in compositions intended for inhalation.

Example 11

500 mg of beclomethasone was dissolved in ethanol, was added to 50 ml of raw slurry CSA (10% wt./about.) and was spray dried under the conditions described in example 10. The size of the microcapsules obtained by this method was determined in non-aqueous system, as described in detail in example 10. The microcapsules had an average size of 3.13 of 0.71 μm, and 90% of the mass was in the range of 2 to 5 microns.

Beclomethasone was extracted from the particles by deposition of CSA in 10% trichloroacetic acid, and the supernatant was extracted with ethanol. The extract in ethanol were analyzed by GHWR at a wavelength of 242 nm. Found in this extract beclomethasone was present in the free state, but when was extracted with albumen residue, they found the presence of beclomethasone connected with native CSA. It was found that, despite the fact that the largest part of the active compounds were free, some were present in SV is twice control over the release of the active compound over an extended period of time compared to the unbound drug.

Example 12

At that time, as at least in examples 10 and 11, any binding of the active compounds was due to the natural properties of albumin, in this example, receive the product after the initial cross-linking active compounds, prior to spray drying.

To a solution of methotrexate 10 mg/ml) was added 25 mg of carbodiimide (EDCI). This solution was stirred for 4 hours to initiate and ensure complete activation of methotrexate. To an activated drug was added 50 mg CSA and was stirred for 3 hours at room temperature. Methotrexate chemical were associated with CSA through the amino group of the albumin. This conjugate is then used as raw material for spray drying as described in example 10.

From the thus obtained microcapsules were sampled, to determine their characteristics and analyzed for the content of drugs. The microcapsules had an average size of 3.2 0.6 microns, and 90% of the mass was in the range of 2 to 5 μm. The analysis of the content of drug in the microcapsules showed that the microcapsules are not freed medicine; even after the dissolution of the drug was still associated with CSA. Cleavage of albumin by proteinase K was released associated in addition to the species. Previously it was shown that the activity of doxorubicin attached to polymeric carriers, has a beneficial effect on the tumor, demonstrating a phenotype resistant to many drugs.

Example 13

Microcapsules of naproxen were made as described in examples 10 and 12, using the ratio of drug to CSA from 1 to 5.

Soluble microcapsules remained active connection nonaqueous solvent. Moreover, the dissolution of the microcapsules in an aqueous solution of the active compound remained bound to albumin, which is confirmed by the analysis by GHWR at a wavelength of 262 nm, as before.

Naproxen was free from albumin by splitting the proteinase K and esterase.

Example 14

Using samples of microcapsules produced in examples 8 to 13 were evaluated by their behavior in the inhaler, spray, dry powder. Reproducible dosing of each composition was evaluated in conjunction with the behavior of the sample by sputtering using a microscopic technique.

A sample of each composition was added to the tank funnel experimental inhaler, spray dry powder (COI). E is O. This measuring device was calibrated using spray dried lactose.

Despite the fact that the quantity measured in the dosing device, different samples was changed, as a function of their composition, the reproducibility of the dosing for each sample was constant; the average for the three doses of 0.25 to 5.0 mg. sample Behavior when spraying studied by connecting the nebulizer to the vacuum chamber; imitation inhalation was achieved by release of the vacuum through the COI, and collecting the released dose was carried out on a glass slide, covered with resin. These slides were evaluated on the dispersion of particles. They showed that the COI was deagglomerated samples, forming a uniform dispersion of microparticles on slides.

Example 15

Characteristics of dry powder compositions of examples 10 to 13 were analyzed using the dual impinger (Apparatus A for inhalation under pressure, the British Pharmacopoeia 1988) after discharge from a Rotahaler (Glaxo UK), 7 ml in stage 1 and 30 ml in stage 2 of distilled water. The composition was delivered from gelatin capsules N 3 using the Rotahaler connected to the dual impinger through the rubber adapter. Vacuum pump servant who was stiglo of the level 1 and 2 impinger. All samples showed the largest share of deposits in stage 2 impinger, indicating that it is optimal for delivery to the alveoli particle size.

Example 16

A comparison was made between dosing and fixed deposits of insoluble and soluble microcapsules microcapsules obtained as described in example 10, the lungs of rabbits.

Anesthetized new Zealand white rabbits were injected soluble microcapsules or fixed microcapsules. Dosing was carried out using a computer controlled aerosol inhaler (Mumed Ltd., UK). Soluble microcapsules suspended in CFC 11 and the fixed particles suspended in water. After a dose of the lungs of rabbits were removed and evaluated the deposition of capsules.

It was found that the fixed capsule remained intact in the alveolar tissue of the lung. This suggests that the microcapsules are suitable for dispersion in lung size. For comparison, there was no presence of intact soluble microcapsules; these capsules are dissolved in the fluids of the lung. However, when using fluorescent microscopy was observed the presence of adduct FITZ/lysine in some part of the alveolar dannym capsules, which was not there.

1. Microparticles of water-soluble material, which are smooth and spherical, and at least 90% of which have a mass-average particle size of 1 to 10 μm, for use in therapy or diagnosis.

2. Microparticles of a mixture of water-soluble therapeutic or diagnostic material and other water-soluble material, and the particles are smooth and spherical, and at least 90% of which have a particle size of 1 µm and 10 µm.

3. Microparticles under item 2, characterized in that they are obtained by spray drying an aqueous solution of the specified water-soluble material and a therapeutic or diagnostic agent.

4. Microparticles according to any one of the preceding paragraphs, characterized in that the particles have dimensions of 1 to 5 microns.

5. Microparticles according to any one of the preceding paragraphs, characterized in that they have a maximum interquartile scope 3 microns.

6. Microparticles under item 5, characterized in that they have a maximum interquartile scope 2 microns.

7. Microparticles according to any one of the preceding paragraphs, characterized in that they are sterile.

8. Microcosim in water material.

9. Microparticles according to any one of the preceding paragraphs, characterized in that they additionally have to bind with receptors in the alveoli component.

10. Microparticles according to any one of the preceding paragraphs, characterized in that a water-soluble material is a carbohydrate.

11. Microparticles according to any one of paragraphs.1 to 9, characterized in that a water-soluble material is amino or polyaminoamide.

12. Microparticles according to any one of paragraphs.1 to 9, characterized in that a water-soluble material is a fatty acid or its ester.

13. Microparticles according to any one of paragraphs.1 to 9, characterized in that a water-soluble material is a protein, peptide or enzyme.

14. Microparticles under item 13, wherein the water-soluble material is a human protein or its fragment, natural or recombinant form.

15. Microparticles under item 14, characterized in that a water-soluble material is human serum albumin.

16. Microparticles according to any one of the preceding paragraphs, characterized in that the water-soluble material for the formation of microparticles has been chemically or enzymatically modified.

17. Strlocal therapeutic agent in the form of microparticles according to any one of the preceding paragraphs.

18. Medicinal product for administration via the respiratory tract, containing a therapeutic agent, wherein therapeutic agent is an microparticles according to any one of paragraphs.1 - 16.

19. A method of treating disease by injecting the patient an effective amount of a therapeutic agent, which has an impact when introduced through the respiratory tract for the treatment of this disease, including the introduction of therapeutic agent in the form of microparticles according to any one of paragraphs.1 - 16.


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