Therapeutic preparation for inhalation, the method of its production (options)

 

(57) Abstract:

Use: medicine for the treatment of diabetes. Therapeutic drug contains insulin and a substance which enhances the absorption in the lower respiratory tract. It is made in the form of a dry powder suitable for inhalation. The powder contains at least 50 wt.% insulin and substances that increase its absorption, in the form of particles with a diameter up to 10 microns or agglomerates of such particles. The powder may further comprise a pharmaceutically acceptable carrier. Insulin may be insulin bull, pig, biosynthetic or synthetic human insulin or a biologically active derivative of human insulin. A substance that enhances the absorption of insulin in the lower respiratory tract - surfactant, bile salt-derived bile salts, sodium salt ursodeoxycholate, taurocholate, glycocholate, eurodiversity, phospholipid, Alkylglucoside and other Drug can be in the device for inhalation. The method of obtaining the drug: get solution of insulin and a substance enhancing its absorption in the lower respiratory tract, the solvent is removed to produce a powder. The powder is at least 50% of celofane specified powder. the 2nd variant of the method. Mix the dry insulin with a dry substance that enhances its absorption. Get the powder, which is at least 50% consists of particles with a diameter of 30 μm. Can be obtained first mixture, which is then pulverized and/or mix to obtain the powder. Mixing can be carried out in the presence of the media. The invention allows to reduce the inconvenience and discomfort in the treatment of patients with insulin therapy. 3 C. and 38 C.p. f-crystals, 9 Il., 3 table.

The invention relates to a therapeutic preparation of insulin, which is suitable for inhalation.

Insulin plays a Central role in the regulation of carbohydrate, fat and protein metabolism in the body. Diabetes mellitus (commonly referred to as just diabetes) is a disease that is characterized by impaired regulation of metabolism, particularly glucose metabolism. Animals in the normal state, the increase of glucose in the blood, which occurs immediately after eating, stimulates the beta cells of the pancreas to excrete insulin, a peptide hormone, into the bloodstream. Insulin binds to insulin receptors on several cell types, particularly in mysen is keeping blood glucose returns to normal, the morning level, the amount of insulin in the blood also decreases. In the absence of insulin, the glucose in the blood is rising to dangerously high levels (a condition called hyperglycemia), leading to the result may be fatal. Too high amounts of insulin causes an abnormally low glucose (hypoglycemia), which is also dangerous and can be fatal. The specimens in good condition inherent feedback makes the cycle, regulating insulin secretion and removing it from the big circle of blood circulation, and prevents both hyperglycemic and hypoglycemic events.

Diabetes is a disease that affects about 3% of the Swedish population. Of these 3% to about 20% suffer from diabetes type I, and the rest from type II diabetes.

Diabetes type I, or insulin-dependent diabetes mellitus (IDDM) usually begins in childhood. It is characterized by atrophy of the beta cells of the pancreas that results in reduction or termination of production of insulin and makes the patient dependent on insulin from outside to survive.

More common type II diabetes, or insulinsee, first, to have normal or even high levels of insulin in the blood, but they found abnormally low rate of cellular uptake of glucose in response to insulin. Although type II diabetes can often be treated by controlling the diet of the patient, the introduction of exogenous insulin in addition to insulin secreted by the beta cells of the patient, it may also be necessary.

Insulin cannot be administered orally in effective doses, as it is quickly destroyed by enzymes in the gastrointestinal tract and low pH in the stomach before it reaches the blood stream. The standard method of administration is subcutaneous injection of isotonic insulin, usually by the patient himself. The need for injections creates a lot of inconvenience and discomfort for many patients, and the injection may be a local reaction. In addition, there is an abnormal nefiziologichnoe profile in plasma concentrations for injectable insulin. Such abnormal profile in plasma concentrations is undesirable and increases the risk of side effects associated with prolonged time of diabetes.

Due to these disadvantages, there is a need for insulin in formano many proposals. For example, the proposed products for nasal, rectal and buccal injection, and the main efforts are concentrated on products for nasal administration. Nasal introduction, however, problematic and allows only low biological availability. Recently, a growing interest in the delivery of systemically acting drugs via the lungs, and some studies include pulmonary delivery of insulin. Most of these studies refers to solutions or suspensions for pulmonary delivery, for example, with the help of nozzles under pressure dosing inhalers, and in all cases with limited success.

The invention

Presently discovered that insulin may be included in a dry powder preparation for inhalation containing the substance which enhances the absorption of insulin in the lungs, of such drug, insulin may be absorbed with therapeutically acceptable speed and therapeutically acceptable amount. Increases absorption" means that the amount of insulin absorbed into the pulmonary circulation, in the presence of the amplifier is higher than the number of insulin, absorber what is therapeutic drug contains active compounds (A) insulin and (B) a substance which enhances the absorption of insulin in the lower respiratory tract, and this drug has the form of a dry powder suitable for inhalation in which at least 50% of the total mass of active compounds consists of (a) primary particles having a diameter less than 10 microns, for example, from 0.01 to 10 μm, preferably from 1 to 6 microns, or (b) agglomerates of the above-mentioned particles.

Therapeutic drug of the present invention may contain only mentioned active compounds, or it may contain other substances such as pharmaceutically acceptable carrier. Such media may consist largely of particles having diameter less than 10 μm, so that at least 50% of the resulting powder consists of optional agglomerated primary particles having a diameter less than 10 μm, on the other hand, the carrier may consist largely of larger particles ("large particles"), so "ready-to-use mixture can be composed of active compounds and the above-mentioned media. In ready-to-use mixture, known, on the other hand, as an interactive or adhesive mixture, small chastise on the surface of large particles excipient (in the present invention pharmaceutically acceptable carrier). In this case, it is preferable that the active compounds were not in the form of agglomerates before the formation of an ordered mixture. Large particles may have a diameter greater than 20 μm, such as more than 60 μm. Near the lower limits of the diameter of the large particles is not critical values, so can be used particles of different sizes, if desired, in accordance with the specific requirements of the specific wording. In ready-to-use mixture to large particles are not required to have the same size, but may be beneficial to within an ordered mixture of large particles were of similar size. Preferably, the larger particles have a diameter of 60-800 mm.

Therefore, in a particular variant of its implementation of the present invention provides a therapeutic preparation of insulin and a substance which enhances the absorption of insulin in the lower respiratory tract, and this drug is in the form of a dry powdered preparation suitable for inhalation in which at least 50 wt.% be (a) particles having a diameter less than 10 microns, or (b) agglomerates of the above-mentioned particles; in another specific embodiment, this invention provides therapeu ways, and pharmaceutically acceptable carrier, and this drug is in the form of a dry powder suitable for inhalation in which at least 50 wt.% be (a) particles having a diameter less than 10 microns, or (b) agglomerates of the above-mentioned particles; and in yet another specific embodiment, the present invention features a therapeutic drug containing the active compounds (A) insulin and (B) a substance which enhances the absorption of insulin in the lower respiratory tract, with at least 50% of the total mass of active compounds (A) and (B) consists of particles having diameter less than 10 microns, and a pharmaceutically acceptable carrier, and this drug is in the form of a dry powder drug suitable for inhalation in which an ordered mixture can be made between active compounds and a pharmaceutically acceptable carrier.

Preferably, at least 50 wt.%, even at least 70 wt.% or at least 80 wt.%, and preferably at least 90 wt.% from the total mass of active compounds (A) and (B) are particles with diameter less than 10 microns, or agglomerates of such particles, and, when dry powdered drug contains media other than in the case when chelation at least 80 wt.%, preferably 90 wt.% from the total mass of the dry powder comprises particles having a diameter less than 10 microns, or agglomerates of such particles.

Although dry powder for inhalation, with or without a pharmaceutically acceptable carrier may contain agglomerates of particles, as indicated above, during inhalation any agglomerates must be desagglomeration with the formation of a powder in which at least 50% of particles having a diameter up to 10 microns. The agglomerates may be the result of a process variable agglomeration, or they may simply be the result of close contact of the powder particles. In any case it is essential that the agglomerates can be desagglomeration, for example, by mechanical means in the inhaler, or otherwise, in the above-mentioned particles. In General, it is preferable that the agglomerates were not formed in ready-to-use mixture. In the case of ready-to-use mixture of active compounds should come from large particles during inhalation, or by mechanical means in the inhaler, or simply under the action of inhalation, or other means, and an active connection then deposited in the lower respiratory tract, and the particles of the carrier - mouth.

A substance which enhances the absorption of insulin in the lungs, called here by the amplifier may be any of those compounds which are increasing absorption through the layer of epithelial cells in the lower respiratory tract in PR is s.

(1) Strengthening paracellular penetrating ability of insulin by inducing structural changes in tight joints between the epithelial cells.

(2) Strengthening transcellularly penetrating ability of insulin by interacting with extracting protein or lipid components of the membrane, and destruction through this membrane integrity.

(3) the interaction between the amplifier and insulin, which increases the solubility of insulin in aqueous solution. This can occur by preventing the formation of aggregates of insulin (dimers, trimers, hexamers) or by solubilization of molecules of insulin in the micelles of the amplifier.

(4) reduction of the viscosity or dissolution of the mucous barrier lining the alveoli and passages of the lung, due to which the epithelial surface is exposed to direct absorption of the insulin.

The amplifiers can operate only one of the above mechanisms, or two or more of them. The amplifier, which operates through several mechanisms, more eligible for promotion effective absorption of insulin than the amplifier, which uses only one mechanism or two. Example: the eat mechanisms, listed above. Surfactants are amphiphilic molecules containing both lipophilic and hydrophilic part, with the changing balance between these two characteristics. If the molecule is highly lipophilic, low solubility of substances in water may limit its suitability. However, if the overwhelmingly dominated by the hydrophilic portion of the surface-active properties of the molecule can be minimal. Therefore, in order to be effective, the surfactant should bring an appropriate balance between sufficient solubility and sufficient surface activity.

Another property of surface-active substances, which may be important is the total charge of the surface-active substances with pH in the lung (approximately 7,4). Isoelectric pH of the insulin is 5.5. At pH 7.4 insulin has a negative total charge. This results in electrostatic repulsion between the molecules of insulin, which, in turn, prevents aggregation, and thereby increases the solubility. If a surfactant is also negatively charged, and, in addition, can vzaimoot additional repulsion. Therefore, the anionic surfactant will have an additional advantage in comparison with surface-active substances having a neutral or total positive charge at physiological pH) to enhance the absorption, contributing to stabilization of insulin in a Monomeric state.

A number of different compounds potentially useful as amplifiers for the methods of the present invention, is tested on rats, as described below in example 5. Other substances with known properties enhance absorption or physical characteristics that make them suitable for use in the method of the present invention, can be easily tested by professionals in this area of engineering in vivo or, on the other hand, the in vitro assays described in example 6.

It is possible that a combination of two or more substances-amplifiers also gives satisfactory results. The use of this combination in the method of the invention is considered as being within the invention.

The amplifier used in the methods of the present invention, will combine effectively increased absorption of insulin from (1) lack of toxicity when used venom condition. The toxicity of a given substance can be tested by standard methods, such as multi-table analysis, for example, as described in Int. J. Pharm. 65 (1990), 249-259. Properties of powder of a given substance may be inferred from information published about this substance, or empirically.

One very promising type of amplifier are salts of fatty acids. It is established that the sodium salts of saturated fatty acids with a chain length of 10 carbon atoms (i.e., caprint sodium), 12 carbon atoms (sodium laurate) and 14 carbon atoms (monistat sodium) fit well in the method of the present invention. Discovered that the potassium salt and lysine salt of capric acid are also effective in the method of the invention. If the length of the carbon chain of less than 10 atoms, the surface activity of the supercial-active substances may be too low, and if the length of the carbon chain than 14 atoms, the low solubility salts of fatty acids in water limits its application.

For the present invention the most preferred substance which enhances the absorption of insulin in the lower respiratory tract, is caprint sodium.

Therefore, in a particularly preferred embodiment, on the - aprint sodium, in the form of a dry powder suitable for inhalation in which at least 50% of the total mass of active compounds (A) and (B) are (a) primary particles having a diameter less than 10 microns, for example, from 0.01 to 10 μm, preferably from 1 to 6 microns, or (b) agglomerates of such particles; specifically, in a particularly preferred variant of its implementation of the present invention offers:

- therapeutic preparation of insulin and caprinate sodium, and mentioned the drug is in the form of a dry powder suitable for inhalation in which at least 50 wt.% be (a) particles having a diameter less than 10 microns, or (b) agglomerates of the above-mentioned particle;

therapeutic preparation containing insulin, caprint sodium and a pharmaceutically acceptable carrier, and the said preparation is in the form of a dry powder suitable for inhalation in which at least 50 wt.% be (a) particles having a diameter less than 10 microns, or (b) agglomerates of the above-mentioned particles; and

therapeutic drug containing the active compounds (A) insulin and (B)-caprinate sodium, with at least 50% of the total mass of active compounds (A) and (B) are particles with diameter less than 10 μm, and formats is galazii, in which of the active compounds and a pharmaceutically acceptable carrier can be formed ready-to-use mixture.

Various contrini can change the solubility of salts of a saturated acid in water, so that the amplifier containing a carbon chain other than 10-14 atoms, is more preferable than the amplifiers, which are defined here above. Salts of unsaturated fatty acids may also be suitable for the present invention because they are more water soluble than salts of saturated fatty acids, and, therefore, may have a longer carbon chain than the last, and still save solubility needed to successfully enhance the absorption of insulin.

The suitability of the amplifiers in the method of the present invention was tested bile salts and derivatives of bile salts. All tested salts (sodium salt ursodeoxycholate, taurocholate, glycocholate and eurodiversity) effectively increase the absorption of insulin in the lungs.

As amplifiers was also tested phospholipids. Found that single-chain phospholipid (lysophosphatidylcholine) is an effective amplifier, while two ducanh f which can be explained by the fact, what ducane phospholipids significantly less soluble in water than their single-copy; however, it is reasonable to expect that ducane phospholipids with a shorter chain having greater solubility than their long-chain counterparts, will be useful as enhancers in the present invention, so can be used as single-and ducane phospholipids.

As the amplifier of the present invention were tested one glycoside-octylglucoside, and discovered that it has some properties enhance absorption. You can also expect other Alkylglucoside, such as thioglucopyranoside and multipurposed will also find properties increasing uptake in the methods of the present invention.

Cyclodextrins and their derivatives effectively enhance the nasal absorption of insulin and may in a similar way to function in the lungs. In the method of the present invention tested dimethyl - cyclodextrin, and discovered that it has the effect of increasing uptake.

Other potentially suitable surfactants are sodium salicylate, 5-methoxysilyl sodium and surface-active substances occurring in nature, such for example, anionic surfactants described above) nature contrina may be important. Specific selected Kontron can affect the properties of the powder, solubility, stability, hygroscopicity, and local and/or systemic toxicity amplifier or any wording that contains the amplifier. It can also act on the stability and/or solubility of insulin, with which it connects. In General, it is expected that the monovalent cations of metals such as sodium, potassium, lithium, rubidium and cesium, are suitable as contrino for anionic amps. Ammonia and organic amines to form another class of cations, which, as expected, are suitable for use with anionic amplifiers containing integral part of the carboxylic acid. Examples of such organic amines include ethanolamine, diethanolamine, triethanolamine, 2-amino-2-methylethylamine, betaine, Ethylenediamine, N,N-dibenzylethylenediamine, arginine, hexamethylentetramine, histidine, N-methylpiperidin, lysine, piperazine, spermidine, spermine and Tris(hydroxymethyl)aminomethan.

As effective increased absorption of insulin in the lungs was observed for many of the tested amplifiers, it is expected that proves effective increase the bioavailability of insulin, delivered through the nasal membranes, and they are checked as the amplifier in the methods of the present invention. Although it is proven that they are unsuitable for delivery to the lung by as much as used here animal models, it seems that this was mainly due to technical difficulties, and if these difficulties are overcome, it is possible to achieve the successful delivery of the lung by. The chelators are a class of amplifiers, which are believed to act by binding calcium ions. Because calcium ions contribute to the preservation of the space between the cells and, in addition, reduce the solubility of insulin binding of these ions theoretically, as would be the solubility of insulin and would be paracellular penetrating ability of insulin. Although discovered that one tested hematopathy agent - sodium salt of ethylenediaminetetraacetic acid (adtc) is ineffective to enhance the absorption of insulin in experimental rat model, it may be that other binding ions of calcium chelating agents will be more suitable.

In General, it is desirable to keep the ratio of insulin to the amplifier as possible in the, the button to minimize the risk of side effects, both local and systemic, which can be inherent in the amplifier. The optimal ratio of insulin and amplifier can be set for the amplifier by checking different proportions on models in vitro, as described here. For example, insulin can be connected with carinatum sodium in the following mass proportions: 50/50, 75/25, of 82.5/17.5 and 90/10. A significant improvement in the absorption of insulin get with 50% and 25% caprinate sodium: 10% give a slight improvement of absorption, and the results from 17.5% are intermediate. This indicates that the lowest effective concentration caprinate sodium for use in the methods of the present invention is approximately 15-25% and perhaps 20-25%. Other amplifiers may be higher or lower optimal concentration relative to insulin, and therefore, each amplifier must be tested separately. However, based on the above results it is expected that the optimal number of amplifier such as surfactant will generally be in the range of 10-50% of the mixture of insulin with the amplifier, for example, 15-50%, as well as 25-50%. It should be noted that the above-mentioned zootomical or other additive, which can be incorporated, for example, to improve the powder properties of the formulation.

The amount of absorbed insulin according to the present invention can be substantially higher than the amount absorbed in the absence of the amplifier. In the example shown here 4 shows that therapeutic drug of the present invention inhalation detects bioavailability, three times the bioavailability inhalation of the drug from one of the insulin.

Preferably, the amount of insulin absorbed according to the present invention, significantly (p < 0,05) higher than the amount absorbed in the absence of the amplifier.

As indicated here above, additional substances that are normally included in therapeutic drugs, such as pharmaceutically acceptable carriers may be included in therapeutic drug of the present invention. Additional substances may be included, for example, to dilute the powder to a quantity that is suitable for delivery from the particular powder inhaler; to facilitate processing of the formulation, to improve the powder properties of the drug; to improve the stability of the drug, for example, with pomo to have an adverse impact on the stability of insulin or amplifier absorption or adversely affect the absorption of insulin. It must be stable, non-hygroscopic, of having good powder properties and do not have harmful effects on the respiratory tract. As examples of possible additives can be mentioned mono-, di - and polysaccharides, alcohols, sugar and other polyols, such as, for example, lactose, glucose, raffinose, melezitose, lactic, ▫ maltitol, trehalose, sucrose, mannitol and starch. As reducing sugars, such as lactose and glucose, have a tendency to form complexes with proteins, the preferred additives for use in the present invention can be nereguliruemyi sugar, such as raffinose, melezitose, lactic, ▫ maltitol, trehalose, sucrose, mannitol and starch. Depending on the inhaler, which is used, the total amount of such additives may vary in very wide limits. In some cases, requires a small amount of additives or does not need, while, for example, in the case of asthma, requiring large amounts of powder, a very high percentage of a therapeutic compound may consist of additives. The right amount of additives will be easily determined by the expert in this field of technology in accordance with the particular case.

A suitable mechanism is powder. There are many such devices, as a rule, for the delivery of anti-asthma and anti-inflammatory drugs in the respiratory system. The preferred device is an inhaler for dry powder, the construction of which provides for the protection of the powder from moisture and in which there is no risk of filing a random large doses; in addition, if possible, the desired protection of the powder from light; high spray fraction and high deposition in the lungs in a wide range of flow rates; low tolerance dose and sprayed fraction; low latency powder at the tip; low adsorption on the surfaces of the inhaler; the flexibility of dose and low inhalation resistance. The inhaler preferably is dose inhaler, although can also be used mnogorazovye inhalers, preferably such as mnogorazovyj, driven by the breath inhaler reusable dry powder. Preferably used an inhaler is a roller, which is driven by the breath inhaler single dose dry powder.

Described powdered drug can be manufactured in several ways with the connections and when appropriate case (i.e. when no means an ordered mixture), any carrier in a suitable mill, for example, in a slim mill, to obtain a primary particle size in the range suitable for the maximum deposition in the lower respiratory tract (i.e., up to 10 µm). For example, can be mixed in the dry state insulin and powders amplifiers and the media, if appropriate, and then finely grind these substances together; on the other hand, substances can be ground separately and then mixed. When compounds that are mixed have different physical properties such as hardness and brittleness, resistance thin grinding varies, and they may require different pressures for breaking up particles of suitable size. Therefore, when substances are crushed together, the resulting particle size of one of the components may be unsatisfactory. In this case it will be advantageous to grind the different components separately and then to mix them.

It is also possible to first dissolve the components, including when no means ready to use mix media, in a suitable solvent, e.g. in water, to obtain SmartVista, what on nasal absorption of insulin is affected by the pH value of the drug and the absorption increases when moving either up or down from the isoelectric point of insulin, which is about 5.5. However, at a pH substantially above or below 5.5 insulin may be less stable and, in addition, must be taken into account pharmaceutical adopted limits for inhalation products from pH 3.0 to 8.5, so a product with a pH outside of these limits may cause irritation and reduction of the respiratory tract. To get the powder, the solvent must be removed in a way that retains the biological activity of insulin. Suitable drying methods include the concentration in vacuum, open air drying, spray drying and drying by freezing. Avoid temperatures above 40oC with exposure time longer than a few minutes, as there can be some destruction of the insulin. After the stage of drying of the solid material, if necessary, can be crushed to produce large powder, and then, if necessary, finely chopped.

If it is desired fine powder before putting it in a suitable inhalation device can be redesigned to improve flow characteristics, teristically. In this case, the device should be of such a form that ensures that the agglomerates essentially disagglomerated before leaving the device, so that the particles included in the respiratory tract of a patient, for the most part, have a size within the desired interval.

When it is desirable to obtain ready-to-use mixture, the active compound may be recycled, for example, in micronizer to obtain, if desired, particles with a certain interval sizes. The media can also be recycled to obtain, for example, the desired size and the desired surface properties, such as the ratio of surface area to mass, or a defined surface roughness, and to ensure optimal adhesion strength in ready-to-use mixture. Such physical requirements for ready to use mixture of well known and there are various ways to obtain the ready to use mixture which satisfies the above requirements and can be easily installed by a specialist in this field of technology in accordance with the specific conditions.

The invention is described by way of examples, which are intended to illustrate, but not limit the scope of the invention.

Add lactose (available commercially, 9.2 grams) and again bring the pH to 7.4. The solution is stirred until a transparent state or to a state of light opalescence and concentrated by evaporation at a temperature of 37oC for two days.

The obtained solid sintered material is crushed and sieved through a sieve of 0.5 mm, the resulting powder is pulverized in a jet mill to a particle size of about 2 microns.

Example 1

therapeutic preparation of insulin and caprinate sodium, the ratio of 75:25

In chemical beaker add a semisynthetic human insulin (9.75 g) and water (250 ml). To dissolve the insulin lowers the pH to 3.4 1 M HCl, and then raise the pH to 7.4 1 M NaOH.

Add caprint sodium (Sigma, 3.25 g) and again bring the pH to 7.4. The solution is stirred until it becomes transparent or slightly opalescent, and concentrated by evaporation at 37oC for two days.

The obtained solid sintered material is crushed and sieved through a sieve of 0.5 mm, the resulting powder is pulverized in a jet mill to particles with a diameter of about 2 microns.

Example 3

therapeutic preparation of insulin and caprinate sodium lactose, a ratio of 4:4:92

Follow the procedure of example 2, using 0.5 g of semisynthetic human insulin, 150 ml of water, 0.5 g caprinate sodium and 11.5 g of lactose.

Studies inhalation

Study 1

Using the product of example 1 for inhalation studies on two dogs. The drug is loaded into the device for inhalation Wright Dust Feed and administered to dogs. The dose level is IU/kg (IU = one unit of human insulin = 35 µg of human insulin, 100%). Measure the performance of blood glucose and insulin in plasma over different periods of time and summarize the results in tables 1 and 2.

The tables show that the formulation of insulin with carinatum sodium significantly increases the level of insulin in the plasma and reduces the concentration of glucose in the blood. The maximum rate for insulin in the plasma and the minimum rate for glucose in the blood are reached in approximately 15 and 60 minutes, respectively.

Investigated the Arata inhalation Wright Dust Feed at a constant level of dosage IU/kg Determine the effect of each formulation on the content of insulin in plasma and blood glucose through different time periods and the results show in Fig. 1 and 2. Found that while the control formulation containing no amplifier, essentially, does not change the content of insulin in the plasma, the formulation containing both insulin and the amplifier increases the amount of insulin in plasma from 20 MK U/ml at zero time up to 80 MK U/ml after 15 min after inhalation of the powder. Similarly, control animals registered maximum drop of glucose in the blood of about 0.5 mmol/l after inhalation of insulin without the amplifier, while in animals, which are inhaled insulin with the addition of the amplifier, register a temporary drop of about 1.7 mmol/l 4.0 mmol/l to 2.3 mmol/L. Thus, insulin in combination with amplifier - carinatum sodium is rapidly absorbed and eliminated from the systemic circulation with the corresponding decrease in the content of glucose in the blood. Conversely, insulin with the carrier (lactose), but without the amplifier is absorbed, how it is possible to detect only a very small extent (p = 0.0002 for insulin with carinatum against insulin with lactose).< two dogs. The introduction of drugs is carried out using the apparatus for inhalation Wright Dust Feed. Through different periods of time after inhalation measure the content of insulin in plasma and blood glucose. The results are shown in Fig. 3-6 show that insulin in combination with carinatum of sodium at different ratios is absorbed quickly and that the maximum value get in 20-30 minutes, accompanied by a corresponding fall of glucose in the blood. The results also show that inhalation powder insulin can be obtained plasma profile that is more similar to natural physiological profile than the profile obtained after subcutaneous injection of insulin.

Example 4

Insulin and caprint sodium 75:25 mixture of finely ground powders

Biosynthetic human insulin (53 g) are ground in a jet mill Airfilco Jet Mill (trade mark Airfilco Process Plant Ltd.) under nitrogen pressure (supply pressure 7 bar, the pressure in the chamber 5 bar) to the primary average particle diameter of 2.4 μm.

Caprint sodium (170 g) are ground in a jet mill Airfilco Jet Mill (TM) under nitrogen pressure (supply pressure 5 bar, the pressure in the chamber 3 bars) to a primary average particle diameter of 1.6 μm.

the om in accordance with the following procedure. Half of the insulin loaded into the mixing installation comprising a mixing chamber with a volume of 4.4 l, separated by a sieve with mesh size of 1 mm for two compartment with a metal ring in each Department to facilitate the mixing and stirring. Add caprint sodium and the rest of insulin. The mixing chamber is closed, turn 180 degrees and fixed in motorized device for shaking. Turn on the engine and continue shaking for approximately two minutes before until insulin and caprint sodium will not pass through a sieve. Switch off the engine and the mixing chamber is turned 180 degrees, again fixed on the device for shaking and again carry out to shake until all the powder does not pass through the sieve. This procedure is repeated eight times to the total time the shaking was approximately 20 minutes

Thus obtained the drug is administered by inhalation 5 dogs, using the device for inhalation Wright Dust Feed at the dose level IU/kg, and over different periods of time after inhalation determine the content of insulin in plasma.

The obtained results are compared with the contents of the Insa is insulin, crushed as described above, to a primary average particle size of 2.4 μm, and the content of insulin in the plasma, obtained by the introduction of five dogs in the same way and with the same dosage levels described above, therapeutic preparation of insulin and caprinate sodium at a ratio of 90:10. In this case, therapeutic drug is prepared as follows. Semi-synthetic human insulin is subjected to gel filtration to reduce the content of zinc from 0.52% to 0.01% with respect to the content of insulin. Insulin (4.5 g) and caprint sodium (0.5 g) dissolved in water (232 ml). The solution is stirred until it becomes transparent, and adjusted pH to 7.0. The solution is concentrated by evaporation at 37oC for two days. The obtained solid sintered material is crushed and sieved through a sieve of 0.5 mm, the resulting powder is pulverized in a jet mill to particles with an average primary diameter of 3.1 μm.

The results of this comparison are given in Fig. 9. The results show some improvement in the bioavailability of insulin for the formulation of 90:10 and the effective improvement of bioavailability of insulin for drug 75:25 of the present invention when compared with a single insulin (p = 0,0147 to distinguish between 75:ut on rat model for its ability to enhance the absorption of insulin and thus affect the level of glucose in the blood, use various forms of insulin: recombinant or synthetic human insulin or of an ox. Each composition prepared as described above in examples 1-3, drying and treating the solution with insulin amplifier, to obtain a powder which can be inhaled.

The powder is administered to rats by inhalation and then control the level of blood glucose in rats. These levels are compared with the corresponding values obtained for rats, which is administered by inhalation formulations of insulin without the amplifier.

Example 6

The choice of amplifiers

To assess the ability of various compounds amplifiers promote the transfer of insulin and other markers through the monolayer of epithelial cells, conduct standard tests in vitro, using the line of epithelial CaCO-2 cells (available from American Type Culture Collection (ATSS), Rockville, MD, USA) as a model of a layer of epithelial cells that functions in a lightweight, separating the alveoli from the pulmonary circulation. During these tests the amplifier and insulin or other marker is dissolved in aqueous solution at different ratios and concentrations and applied to the apical side of the cell monolayer. After incubation for 60 min at 37oC and 95%ENEA marker, tagged with radioactive isotopes. For a given amplifier (caprinate sodium) tested in the experiments shown in Fig. 5 and 6, the number of the marker (mannitol, 360 mm), which is basolateral side, depends on the concentration of the used amplifier until at least 16 mm caprinate sodium (Fig. 7). This is true even when the mixture of the amplifier with mannitol added insulin (caprint sodium : insulin = 1:3 by weight) (Fig. 8). Also found that this concentration caprinate sodium (16 mm) promotiom absorption of insulin through the cell monolayer. Insulin, which passes through the monolayer in the presence of 16 mm caprinate sodium, doubled compared to the amount of insulin in the absence of any amplifier. It is expected that at higher concentrations caprinate sodium permeability of the cells would grow even further; however, the possible cytotoxicity caprinate sodium may hinder the application of significantly higher concentrations of this particular amplifier.

This model of the in vitro permeability of epithelial cells can be used as a verification tool for rapid testing of the desired amplifier prigodnaya fact, that it additionally contains a substance which enhances the absorption of insulin in the lower respiratory tract, and is made in the form of a dry powder suitable for inhalation in which at least 50 wt.% total insulin and substances that increase its absorption, consists of particles having a diameter up to 10 microns or agglomerates of such particles.

2. Therapeutic drug under item 1, characterized in that it additionally contains a pharmaceutically acceptable carrier.

3. Therapeutic drug under item 2, characterized in that the carrier consists of particles having a diameter up to 10 microns, and at least 50% of the specified dry powder consists of particles having a diameter up to 10 microns or agglomerates of such particles.

4. Therapeutic drug under item 2, characterized in that the carrier consists of coarse particles with a diameter of more than 20 μm, and it is this that allows to form an ordered mixture of insulin, substances that increase its absorption, and media.

5. Therapeutic drug under item 3, characterized in that at least 50% of the particles of the dry powder has a diameter of 1 to 6 μm or represents the agglomerates of such particles.

6. therapeutic composition under item 1 or 4, characterized in that p is of the EPR between 1 - 6 ám.

7. Therapeutic drug according to any one of paragraphs.1 - 6, characterized in that the insulin is an insulin bull, pig, biosynthetic or synthetic human insulin or a biologically active derivative of human insulin.

8. Therapeutic drug under item 7, wherein the insulin is a semi-synthetic human insulin.

9. Therapeutic drug under item 7, wherein the insulin is a biosynthetic human insulin.

10. Therapeutic drug according to any one of paragraphs.1 to 9, characterized in that the substance which enhances the absorption of insulin in the lower respiratory tract, is a substance that enhances the absorption of insulin through the layer of epithelial cells of the lower respiratory tract in the adjacent pulmonary vascular network.

11. Therapeutic drug under item 10, characterized in that the substance which enhances the absorption of insulin in the lower respiratory tract, is a surface-active substance (surfactant).

12. Therapeutic drug under item 11, wherein the surfactant is an anionic surfactant.

13. therapist is the ways, is a bile salt or derivative bile salts.

14. Therapeutic drug under item 10, characterized in that the substance which enhances the absorption of insulin in the lower respiratory tract, is a sodium salt ursodeoxycholate, taurocholate, glycocholate or eurodiversity.

15. Therapeutic drug under item 10, characterized in that the substance which enhances the absorption of insulin in the lower respiratory tract, is a sodium salt taurocholate.

16. Therapeutic drug under item 10, characterized in that the substance which enhances the absorption of insulin in the lower respiratory tract, is a phospholipid.

17. Therapeutic drug under item 10, characterized in that the substance which enhances the absorption of insulin in the lower respiratory tract, is Alkylglucoside.

18. Therapeutic drug under item 10, characterized in that the substance which enhances the absorption of insulin in the lower respiratory tract, is a cyclodextrin or its derivative.

19. Therapeutic drug under item 10, characterized in that the substance which enhances the absorption of insulin in the lower respiratory tract, is a salt ishana of insulin in the lower respiratory tract, is acylcarnitine.

21. Therapeutic drug under item 1, characterized in that it contains sodium salt taurocholate as substances that enhance the absorption of insulin in the lower respiratory tract.

22. Therapeutic drug according to p. 21, characterized in that it additionally contains a pharmaceutically acceptable carrier.

23. Therapeutic drug under item 22, wherein the carrier comprises particles having a diameter up to 10 microns, such that at least 50% of the specified dry powder consists of particles with a diameter up to 10 microns or agglomerates of such particles.

24. Therapeutic drug according to p. 22, characterized in that the carrier consists of coarse particles with a diameter of more than 20 μm, and it is this that allows to form an ordered mixture of insulin, substances that increase its absorption and media.

25. Therapeutic drug according to any one of paragraphs.1 to 24, characterized in that the ratio of insulin and substances that increase its absorption in the lower respiratory tract, is in the range 9 : 1 to 1 : 1.

26. Therapeutic drug for p. 25, characterized in that the ratio is in the range 4 : 1 to 2 : 1.

27. Therapeutic drug under item 2 is the Bohm PP.2 - 5, 7 - 20, 22 - 27, characterized in that the pharmaceutically acceptable carrier is selected from the group comprising mono-, di - and polysaccharides, sugar alcohols and other polyols.

29. Therapeutic drug for p. 28, characterized in that the carrier is an unrestored sugar.

30. Therapeutic drug under item 29, wherein the carrier is selected from the group including raffinose, melezitose, lactic, ▫ maltitol, trehalose, sucrose, mannitol or starch.

31. Therapeutic drug according to any one of paragraphs.1 to 30, characterized in that it is in the device for inhalation, providing protection from moisture and dosing.

32. Therapeutic drug under item 31, wherein the inhalation device is a disposable inhaler single dose dry powder, which is driven by the wind.

33. Therapeutic drug under item 31, wherein the inhalation device is a reusable nebulizer multiple applications for dry powder, which is driven by the wind.

34. Therapeutic drug according to any one of paragraphs.1 - 33 for the treatment of diabetes.

35. The way to obtain therapeutic Ave is ivyshim its absorption in the lower respiratory tract, removing the solvent to obtain a powder, which is at least 50% consists of particles having a diameter up to 10 microns, or solid product, which is then pulverized and/or mix to obtain the powder.

36. The method according to p. 35, characterized in that the solvent is removed by evaporation.

37. The method according to p. 35, characterized in that the solution further add pharmaceutically acceptable carrier.

38. A method of obtaining a therapeutic preparation containing insulin, characterized in that it comprises dry mixing insulin with a substance that strengthens his absorption in the lower respiratory tract, from gaining powder, which is at least 50% consists of particles having a diameter up to 10 microns or mixture, which is then pulverized and/or mix to obtain the powder.

39. The method according to PP.35 to 38, characterized in that it includes an additional stage fine grinding of the specified powder.

40. The method according to p. 38, wherein the mixing is carried out in the presence of a pharmaceutically acceptable carrier.

41. The method according to p. 37 or 40, characterized in that the obtained powder is mixed with pharmaceutically acceptable wear is R>
24.06.93 on PP.1 - 34;

04.02.94 on PP.35 - 41.

 

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