Therapeutic preparation for inhalation

This invention relates to a pharmaceutical composition containing insulin suitable for inhalation. >

Without insulin, glucose levels in blood would be dangerously high (condition called hyperglycemia) and could cause death. Too much insulin generates an abnormally low level of hypoglycemia, which is also dangerous and can also be fatal. In a healthy person, an integrated response system that controls insulin secretion and its cleansing from the cardiovascular cycle that prevents hyperglycemia or hypoglycemia.

Diabetes mellitus is a disease that affects approximately 3% of Sweden's population. Of this 3% era approximately 20% who suffer from type I diabetes and others of type II diabetes.

Type I diabetes mellitus, or insulin-dependent diabetes mellitus (IDDM), appears above the threshold. Ηύη is characterized by shrinkage in the pancreas, which results in a reduction or suspension of insulin production, and makes the patient highly dependent on external insulin to live.

More common type II diabetes mellitus, or insulin-dependent diabetes mellitus (NIDDM), which usually occurs in patients who are older than 40 years of age. These patients may, at least initially, have normal or even high levels of insulin in the blood, but show abnormally low levels of glucose uptake as insulin responses. Although type Π diabetes mellitus can usually be treated by controlling the patient's diet, additional insulin may be required to add to his / her beta cells.

Insulin can not be administered by mouth at effective doses as it rapidly breaks down into the gastrointestinal tract of enzymes and stomach due to low pH levels before it reaches blood transfusions. The usual method of administration is a skin care isotonic solution containing insulin, usually performed by the patient itself. The need for injections causes a lot of discomfort to many sufferers and discomfort and local effects may occur at the injection site. In addition, the pathway of plasma steroids for injected insulin is abnormal and not physiological. This abnormal plasma plasma pathway is undesirable and increases the risk of side effects associated with long-term treatment of diabetes.

Because of these disadvantages, insulin is required in a form that can be given as a drier but with a syringe. Efforts to produce other forms of insulin have led to various ideas. For example, ideas for products for administration of the nose, rectum and mouth, but a great effort has been made to develop products for nasal administration. Nutritional intake is nevertheless subject to difficulty and permits only very low bioavailability. The distribution of drugs that affect the entire body through the lung has been gaining increasing attention in recent years, and some studies have mediated by the spread of pulmonary insulin.

Most people have solutions or suspensions for spreading the lungs with, for example, aerosol and pressure-inhalation doses with limited results.

invention

We have now found that insulin can be combined with a mixture of inhaled powder and a substance that promotes insulin absorption in the lung, but from the mixture the absorption of insulin will be satisfactory from the therapeutic point of view in terms of both speed and volume. "Higher absorption" means that the amount of insulin absorbed into the systemic cycle of the body in the presence of the substance is greater than the amount absorbed, if not present.

This invention is characterized in that a pharmaceutical composition comprising active compounds (A) instillin and (B) a substance that promotes the lower respiratory absorption of the instilin, the mixture is in the form of an inhalation powder suitable for at least 50% of the total body active the compounds consist of (a) primary particles having an age of less than about 10 micrometers, to a diameter between 0.01 and 10 micrometers and between 1 and 6 micrometers, or (b) clusters of said particles.

The pharmaceutical composition of this invention may contain purely active compounds or it may contain other substances, such as a pharmaceutically acceptable carrier. This carrier may consist mostly of particles having a diameter of less than about 10 microns, such that at least 50% of the resulting powder consists of aggregates of potentially collapsible particles having a smaller diameter but about 10 microns; carriers may also consist of substantially larger particles ("coarse particles"), so that "sequences" can form between the active compounds and the carrier. In a series of compositions, also referred to as an interactive or adhesive mixture, finely divided drug data (in the present invention) are distributed fairly over a wide range of carrier materials (in this invention, pharmaceutically acceptable excipient). Preferably, there are physical cases that the active compounds are not in the form of agoa clusters for the formation of the sequestering. The coarse particles can rotate with a diameter greater than 20 microns, such as over 60 microns. Above this lower limit, the diameter of the coarse particles is not very important, so using various grades of coarse particles may be desired as appropriate for a mixture. No conditions apply to the rough part of the mixer, but it may be useful if you dig particle size of the same size within the mixer. The diameter of the coarse particles should be between 60 and 800 microns. The coarse particles can rotate with a diameter greater than 20 microns, such as over 60 microns. Above this lower limit, the diameter of the coarse particles is not very important, so using various grades of coarse particles may be desired as appropriate for a mixture. No conditions apply to the rough part of the mixer, but it may be useful if you dig particle size of the same size within the mixer. The diameter of the coarse particles should be between 60 and 800 microns. The coarse particles can rotate with a diameter greater than 20 microns, such as over 60 microns. Above this lower limit, the diameter of the coarse particles is not very important, so using various grades of coarse particles may be desired as appropriate for a mixture. No conditions apply to the rough part of the mixer, but it may be useful if you dig particle size of the same size within the mixer. The diameter of the coarse particles should be between 60 and 800 microns. but it may be advantageous if you dig part of your age within the mix. The diameter of the coarse particles should be between 60 and 800 microns. but it may be advantageous if you dig part of your age within the mix. The diameter of the coarse particles should be between 60 and 800 microns.

In a particular embodiment of the invention there is provided a pharmaceutical composition comprising an insulin and a substance that enhances the absorption of insulin into the lower respiratory tract, the pharmaceutical composition being in the form of a mixture of powder suitable for inhalation and comprising at least 50% mass of (a) particles of age with which is less than about 10 micrometers (b) clusters of such particles; In addition, the invention includes a further embodiment of a pharmaceutical composition containing insulin, a substance that enhances insulin absorption into the lower respiratory tract, and a pharmaceutically acceptable carrier, the mixture is in the form of an inhalable powder and at least 50% of the mass consist of ) particles less than about 10 micrometres in diameter, or (b) clusters of such particles;

Heist consist of at least 60%, such as at least 70%, at least 80%, and at least 90% of the parent compound of active compounds (A) and (B) of particles within 10 micrometers in diameter, or from the clusters of such particles, and when the powder mixture contains a carrier, which is different from that of a radish mixture, comprises at least 60%, such as at least 70%, at least 80%, and at least 90% healing mass of powdered eyes that are within 10 microns in diameter, from the clusters of such particles.

Although the powder for injection, whether or not containing a pharmaceutically acceptable carrier, may contain particulate clusters as mentioned above, by inhalation of the host should be completely free at clusters, thus at least 50% of Its particles of dust are up to 10 micrometers 1 diameter. The clusters may have been prescribed by a controlled agitation process, or they may have resulted from close contact with the powder. In both cases, it is important that the clusters are broken apart, for example, by mechanical equipment in the container with excessive behavior, thus depending on the value of the particulate matter. The clergy era is generally not known in the series. If a radish mixture is used, the active compounds should be released from the large particles, inhaled by inhalation,

All of the biologically active forms of insulin derived from this invention can be used. For example, bovine, porcine human insulin, produced by whole or in part in living cells, can be used as a biologically active derivative of human insulin ("modified insulin"), which contains, for example, a few amino acid residues as taught by Brange et al In "Diabetes Care" 13: 923,

1990. A modified insulin has been developed to improve various properties, for example, to improve the stability of the process through the pharmacokinetic process (ie a better absorption pathway through the epithelial membrane). The insulin content should be low as zinc reduces insulin solubility and is likely to decrease the rate of absorption and also because zinc oxide may form undesired insoluble dopants with some of the excipients as used in addition to absorption in this invention. Additionally, insulin should be a form of powder that dissolves rapidly in an aqueous solution.

The substance that enhances insulin absorption in the lung, hereinafter referred to as an enhancer, may be one of many compounds that promote the absorption through the lower respiratory tract in the lower respiratory tract and into the adventitious vascular system of the lung. An attacker can handle this in various ways:

(1) With the enhancement of insulin metabolism with the formation of structural changes in the tight bonds between the skin cells.

(2) By enhancing the transfusion of insulin via interactivity at or by withdrawing the protein or lipid component of the membrane and disturbing the integrity of the membrane.

(3) Interactivity enhancement and insulin, which increases insulin solubility in an aqueous solution. This can be done by preventing the formation of insulin clusters (the third, third, hexamera) or by dissolving insulin molecules in adrenal glands.

(4) By reducing viscosity or dissolving the mucus layer surrounding the pulmonary arteries and pelvic lungs, make the surface of the epithelial tissue open for direct absorption of the insulin.

Eykjamir can work according to one or more of the above or two or more. An agent that works in a few ways is more likely to cause an effective absorption of insulin than the one who uses only one or two. Overactive substances are, for example, a category of ewes that are considered to be used by all four of the above mentioned genes. Principal active substances are bilayered molecules that have both lipophilic and hydrophobic components, with different ratios between these two properties. If the molecule is very lipophilic, the low solubility of the substance in its water limits its usefulness. Conversely, if the hydrophilic part is highly dominant, the molecular weight of the molecule is minimized. In order to influence the impact of the surfactant, the right balance between sufficient solubility and sufficient surface activity.

Another feature of surface active substances that may be important is the net loading of the surfactant at the pH of the lung (approximately 7.4). The insulin level of insulin is 5.5. At pH 7.4, the net loading of insulin is negative. This causes electrostatic displacement forces & between molecules of insulin which prevents their aggregation and increases the solubility. If the surfactant is also a negative charge, but it can still be a useful link with insulin, for example, hydrophobic association, the interferon forces between the insulin molecules will be greater. As a result, negative surfactant has added the additional cost (in comparison to those neutral or net-positive at physiological pH) to increase absorption by participating in enhancing insulin monovalent stability.

Many different compounds potentially useful as enhancers in the methods of the invention were tested in rats, as described in Example 5 below. Other substances known to promote absorption or possess chemical properties which are suitable for use in the method of the invention can be readily tested by the mediator in the "vivo" or "possible" vitro analysis of the diagnosis described in Example 6.

It is possible that the combination of two or more enhancers can give a satisfactory result. The use of such compositions in the method of the invention is considered to be the subject of the invention.

Enhers useful in the methods of the invention combine effective boosting insulin absorption, and (1) toxicity in the concentrations used and (2) good duodenal properties, there is a lack of adhesive or waxy solid properties. The toxicity of a particular substance can be tested by standard methods such as MTT testing, for example, as described in Int.

J. Pharm., 65 (1990), 249-259. Duplicate properties of certain content can be verified in published data or in experiments.

One type of egg that promises very good is a salt of fatty acid. It has been found that sodium salts of saturated fatty acids in a carbon chain of 10 carbon long (that is sodium caprate), 12 (sodium chloride) and 14 (sodium mylrate) prove useful in the process of the invention. Potassium and citric salts of capric acid have also proved to be effective in the method of the invention. If the length of the carbon chain is less than about 10, the surfactant activity of the surfactant may be too small and if the length of the chain is greater than about 14, the decreasing solubility of the fatty acid ions reduces water from its utility.

In this invention, it is preferable to use sodium caprate to increase insulin absorption in the lower respiratory tract.

In a particularly preferred embodiment, the present invention provides a pharmaceutical composition comprising a waxy compound (A) of insulin and (B) sodium caprate, suitable for inhalation powder and comprising at least 50% of the total amount of active compounds (A) and (B) of (a) particles of less than 10 micrometers in diameter, for example between 0.01 and 10 micrometers and inclined between 1 and 6 micrometers, or (b) clusters of such particles; This embodiment of the invention typically comprises a pharmaceutical comprising insulin and sodium caprate, the mixture being in the form of a powder suitable for inhalation and at least 50% of the mass (a) particles of less than 10 micrometres in diameter or (b) clusters said particles;

a medicament containing insulin, sodium caprate and a pharmaceutically acceptable carrier, the mixture being in the form of a powder suitable for inhalation and at least 50% of the mass (a) particles of less than 10 micrometres in diameter, (b) colonies of said particles ; and a pharmaceutical composition containing active compound (A) insulin and (B) sodium caprate, wherein at least 50% of the total amount of active compounds (A) and (B) particles less than 10 micrometres in diameter and pharmaceutically acceptable carrier is in the form of an inhalation powder suitable for mixing between the active compounds and the pharmacologically acceptable carrier.

Different intermediates can change the solubility of the salty saturated fatty acid in water, thus increasing the carbon chain other than 10-14, even better than the ones specifically mentioned above. Salts of unsaturated fatty acids may also be useful in this invention as they are water-soluble but salts of saturated fatty acids and may therefore be longer chained than the latter and still retain the solubility that is evident in the absorption of insulin.

The utility of bile salt and the derivative of those enhancing the behavior of this invention was a test subject. All of the ingredients of our test (sodium salts of urethoxycolate, taurocolate, glycocolate and taurodihydrofusidate) effectively absorption of insulin in the lung.

The utility of phospholipid as an enhancer was also tested. revealed that single-chain phospholipid (lysophosphatidylcholine) was actively enhanced, but two two-chain phospholipids (dfoctanoylphosphatidylcholine and didecanoylphosphatidylcholine) were not available. This s

can be explained by the fact that the two-chain phospholipid era will be more volatile in water than the single chain of their lipids; However, it is natural to assume that short chain chains of phospholipids with short chains and greater water solubility than those of their long chains are useful in this invention so that both single and double chain phospholipids may be useful.

The use of glycoprotein, octylgluopyranoside, as an enhancer tested in this invention, was found to have some properties enhancing absorption.

Other alkyl glycosides such as thugoglupyranoside and maltopyranoside also probably have the properties of an enhanced absorption in the methods of the invention.

Cyclodextrin and its derivatives effectively enhance the absorption of nasal insulin and may have a similar effect to the lung. Dimethyl-β-cyclodextrin has been tested by the process of the invention and was found to have an effect on the cytotoxicity.

Other potentially beneficial surfactants of sodium salicylate, sodium 5-methoxysalicylate and natural surfactants such as glycyrrhizinic acid, soaponylcoside and acylkamithine salts.

If the enhancer is an ion (e.g., the negatively charged surfactant described above), the nature of the interaction may be important. The chosen values ​​may affect the properties of powder, solubility, stiffness, water resistance, and local / selective toxicity of the enhancer, or any mixture containing it.

Ηώη can also affect the stability and / or enhance the solubility of the insulin in which it is mixed with a vial. Generally, the power of monovalent metal cations such as sodium, potassium, lithium, rhbidium, and cesium can be expected to be useful as a counterpart for negative-leaked echoes. Ammonium and organic amines form another category of cation that can be expected from power suitable for use with negatively charged vehicles with a carboxylic acid component. Examples of such organic amines are ethanolamine, diethanolamine, triethanolamine, 2-amino-2-methylethylamine, betaine, ethylenediamine, N, D-benzyiethylenetetraamine, arginine, hexamethylenetetraamine, histidine, N-methylpiperidine, Lichine, Pfperazine, Spermidine, Sperm and Tris hydroxymethyl) aminomethane.

Since many of the tested eukars showed an effective enhancement of insulin absorption in the lung, it is assumed that many more will be finer which also have such an effect. microscopic starch enhances the effective bioavailability of insulin that passes through nasal membranes, and were tested as enhancers in the methods of the invention. Although it was found that they had little utility in the distribution of lung, according to the animal model used for the testimonials, it is believed that it was mainly due to technical difficulties which, if solved, can lead to effective pulmonary absorption. Grips are a category of echoes that are believed to be affected by the binding of calcium ions. Since calcium ions contribute to maintaining cellular distance disturbances and also reduce insulin solubility, the binding of these ions would increase the solubility of insulin and increase the flow of insulin between cells. Although a single agent tested for sodium salt of ethylenediaminetetraacetic acid (EDTA) may not be effective in enhancing insulin absorption in the rat model used during the test, other calcium ions-containing adhesives may prove useful.

In general, it is desirable to maintain insulin as well as as high as possible, within the limits that enable rapid and effective increase in insulin absorption. This is important in order to minimize the risk of adverse effects, both local and cardiovascular, attributable to the substance. The desired percentage of insulin can be found and the enhancement of which enhancement by testing various ratios of Jn vivo models, such as those described here. For example, insulin was mixed with sodium caprate in the following w / w ratios: 50 / 50.75 / 25.82.5 / 17.5 and 90/10. A significant increase in insulin absorption was achieved with 50% and 25% sodium caprate; 10% increased absorption was low and nidurstödum for 17.5% was moderate. This indicates that the lowest concentration of sodium caprate which is effective in the practice of the invention is about 15-25% and probably 20-25%. Other enhancers can give higher or lower youthful levels for insulin, so one must test who specifically enhances. On the basis of the above tests, however, one can expect the desired proportion of the level of surfactant content generally in the range of 10-50% of the insulin / additive mixture, for example between 15% and 50% such as between 25% and 50% . It should be borne in mind that the above ratios only represent the proportions of enhancement and insufficiency, and do not take into account the burden of other additives that may be added in, for example, to enhance the powder properties of the mixture. insulin and it must therefore be tested separately. On the basis of the above tests, however, one can expect the desired proportion of the level of surfactant content generally in the range of 10-50% of the insulin / additive mixture, for example between 15% and 50% such as between 25% and 50% . It should be borne in mind that the above ratios only represent the proportions of enhancement and insufficiency, and do not take into account the burden of other additives that may be added in, for example, to enhance the powder properties of the mixture. insulin and it must therefore be tested separately. On the basis of the above tests, however, one can expect the desired proportion of the level of surfactant content generally in the range of 10-50% of the insulin / additive mixture, for example between 15% and 50% such as between 25% and 50% . It should be borne in mind that the above ratios only represent the proportions of enhancement and insufficiency, and do not take into account the burden of other additives that may be added in, for example, to enhance the powder properties of the mixture.

The amount of insulin absorbed according to this invention can significantly differ from the amount of fiber absorbed when no increase is present. Example 4 below shows that the bioavailability of a pharmaceutical composition according to this invention is by inhalation more than three times more than after inhalation of insulin containing insulin alone.

Heist is the amount of insulin absorbed according to this invention significantly (p ≤ 0.05) meiia enviromentic absorption factor of eclipse.

As noted above, additives commonly used for pharmaceutical compositions such as pharmaceutically acceptable carriers may be a part of the pharmaceutical composition of this invention. Additives may be useful in diluting the powder to be suitable for distribution from the powder medicine used; to facilitate the preparation of the mixture; to optimize the properties of the powder; to the best supportedness of the mixture, such as those that warn of quenching or pH-adjusting compounds; or to give the mixture a taste. Additives should not reduce insulin or ocular stability, or interfere with insulin absorption. The pad should also be stable, not hydrated, have good duodenal properties and no adverse effects on the respiratory tract. As a matter of possible additives, mono-, di- and polysaccharides may be mentioned,

A pair of reducing sugars such as lactose and glucose tend to be complex with proteins, are non-reducing sugars such as raffinose, melesitose, lactitol, maltitol, trehalose, sucrose, mannitol and may potentiate a suitable additive for use in this invention. The total amount of these additives used can vary greatly, depending on the type of inhaler used. in some cases there would be no need for a small amount of food, even no additives, but on the other hand, if you use an infusion that requires a large volume of powder, a very high proportion of the drug mixture may consist of additives. The desired amount of additive could be determined simply by a professional at the appropriate site, after each session.

A good thing to do when spreading the powder into the patient's lung is a handy inhaler suitable for use by inhalation of a powder. Many of them are designed specifically for spreading asthma and inflammatory agents through the respiratory tract. Tskid is a powdered powder coated so that the powder is protected from moisture and thus there is no overexposure;

In addition, it is desirable that most of the following elements are present: powder protected from light; high levels of intrinsic particles and high levels of distribution in the lung at the fluid flow rate; small deviation between dose and intestinal part; the amount of powder after the mouthpiece; the top of the dish gives a little bitterness;

dosage may vary; Lftil resisted at home. The inhaler is indicated for single dose although a multidose dose can also be used, known as multi-dose multi-dose air intake actuator activated by inhalation. Heist is the inhaler used as a single dose inhaler, activated by inhalation, powder for use and for single use.

The powder mixture described herein can be produced in a variety of ways, with conventional behaviors. It may be advisable to microfilm the active compounds and any carrier, if applicable (if not intended to obtain a sequestered mixture) in a suitable quinktime, for example, in a fibrillation at any stage of the production cycle, in order to produce primates that are The size of the bill that suits the maximum distribution for reaching the lower part of the respiratory tract (that is less than 10 / <m). For example, you can mix in a dry form of insulin and powder and powder when appropriate and then micro-aggregate; Another option is to flatten the substances individually and mix them together. Since the compounds that blend together possess different chemical properties such as hardness and their leakage, their resistance is different from the microprocessing and can be used to apply varying pressures to breaking them into suitable particle sizes. When micronized together, there may be a risk that the particle levels observed in any of the constituents may be inadequate. In such cases it would be advantageous to emit the different elements individually and then mix them together.

It is also possible to start solving the components together with, in the absence of an aerosol, any carrier in a suitable solvent, such as water, to obtain a mixture at the molecular stage. This method also makes it possible to adjust the pH to the desired amount. Pad is known that insulin absorption is dependent on the pH of the mixture, with increased absorption when increasing the level of insulin for an equilibrium point of about 5.5. However, the insulin may be unstable at pH, especially above or below 5.5, and should also be taken into consideration by the pharmacologically advanced limits at pH 3 to 8.5 for inhalation products, which produce pH at that point Darkness can control irritation and recession in the duct. In addition, powder is obtained by removing the solvent with the behavior of the biological activity of insulin. Suitable drying behaviors your medal of use of underpressure, open drying, drying out and freeze drying. Temperatures above 40 ° C for more than a few minutes should generally be avoided as any metabolism of the insulin may occur. After drying, it may be advisable to grind solids to obtain a rough powder that may be microscopically affected.

If desired, treat the micronized powder more to enhance its flow properties, for example, with a dermatologically distorted image of pruritus particulate clusters that have superior properties for treatment, rather than inserted into the intestinal device that is intended to be used. In this case, the device would be equipped to ensure that the clusters of the particles were completely discharged, leaving them completely apart, leaving the device so that the particles entering the patient's respiratory tract are largely within the desired size range.

As soon as a sequential composition is obtained, the active compound can be treated, for example by microwell, to obtain, on request, particles of a certain size. The carrier may also be treated, for example, to obtain the desired size and surface characteristics, such as a special ratio between surface and weight or certain capability, and to ensure maximum adhesion forces within a series of mixtures. Such substantive requirements are well known in terms of a series of compositions, and various ways are provided for obtaining a series of compositions that meet the above criteria, and which can be easily selected by a person skilled in the art in accordance with circumstances at any given time.

The invention will now be described with examples intended to illustrate, but not limit the scope of the invention.

examples

Samanbnrðardæmi

Medicinal product containing insulin, without any substance

Semi-synthetic human insulin (Diosynth, 03 g) and water (150 ml) were added to a beaker. In order to dissolve the insulin, the pH value was decreased with 1M HCl to pH 3.4 and then raised with 1M NaOH to pH 7.4.

Lactose (prepared, 93 g) was added and the pH adjusted to pH 7.4. The stirring was in solution until it became clear or faint opalescent, it was evaporated by evaporation at 37 ° C for about two days.

The solid cake was pressed and filtered through a 03 mm filter and the powder released was micronized with particles of approximately 2 micrometers in diameter with a jet pellet.

Case 1

Lvfiablanda containing insulin and sodium caprate: f ratio of 75:25.

Semi-fused human insulin (9.75 g) and water (250 ml) were poured into a beaker. In order to dissolve insulin time, the pH value was decreased with 1M HCl to pH 3.4 and then raised with 1M NaOH to pH 7.4.

Sodium caprate (Sigma 3.25 g) was added and the pH value returned to pH 7.4. The solution was stirred until it became taer or faint opalescent and was evaporated by evaporation at 37 ° C over a period of two days.

The solid-fast cake was pressed and filtered through a 03 mm filter and the powder released was micronized in particles with approximately 2 micrometers in diameter with a jet glue.

Example 2

Medicinal product containing insulin and naphtha caprate. with lactose: in the ratio

50:25:25

Half-humanized human insulin (73g) was dissolved in water (150ml) as in Example 1. Sodium caprate (3.75g) and lactose (3.75g) were extracted and the reaction was carried out in Case 1 to produce a powder that consisted mostly of diameters of diameters of about 2 million square meters.

Example 3

A pharmaceutical composition containing insulin and sodium caprate. with lactose: the ratio

4: 4: 92

The reaction was carried out according to Example 2, using 03 g of semi-synthetic human insufficiency, 150 ml of water, 03 g of sodium caprate and 113 g of lactose.

InndBarannsdlmlr

Investigations 1

The mixture according to case 1 was noted in an inhalation study of two dogs. The mixture was placed in the Wright Dust Feed inhaler and given the dogs. The dose level was 1 UTkg (1U. = 1 unit of human insulin = 35pg of human insufficiency, 100%). Blood glucose and plasma insulin were measured at different times and the results shown in Tables 1 and 2 below.

Tafia I

<td>Blood test.time end of access (minutes)</td><td>Blood glucose (Mmol / L)</td><td>The concentration of insulin Qiu / ml)</td>

<td>before</td><td>33</td><td>6.70</td>

<td>03</td><td>3.6</td><td>12036</td>

<td>5</td><td>23</td><td>194.47</td>

<td>10</td><td>23</td><td>19539</td>

<td>20</td><td>money</td><td>139.74</td>

<td>223</td><td>13</td><td>money</td>

<td>31</td><td>2.0</td><td>73.42</td>

<td>45</td><td>1.7</td><td>47.49</td>

<td>593</td><td>1.7</td><td>3631</td>

<td>893</td><td>23</td><td>1938</td>

<td>120</td><td>3.0</td><td>1438</td>

<td>240</td><td>43</td><td>538</td>

fe = not found

Tafia II

<td>Blood sample, from end of access (minutes)</td><td>Glucose in the blood (Mmol / L)</td><td>Insulin strength O / U / ml)</td>

<td>previously</td><td>33</td><td>4434</td>

<td>3</td><td>43</td><td>165.10</td>

<td>6</td><td>43</td><td>15838</td>

<td>12</td><td>33</td><td>money</td>

<td>14</td><td>money</td><td>180.72</td>

<td>19</td><td>3.0</td><td>133.75</td>

<td>30</td><td>2.7</td><td>143.71</td>

<td>45</td><td>25</td><td>9152</td>

<td>60</td><td>2.4</td><td>66.70</td>

<td>90</td><td>2.7</td><td>3858</td>

<td>122</td><td>3.7</td><td>29.15</td>

<td>241</td><td>4.1</td><td>money</td>

<td>2425</td><td>fs.</td><td>19.76 '</td>

fe = not found

Tablets show that the insulin / sodium caprate mixture increases the apparent amount of insulin in plasma and reduces the amount of glucose in the blood. Maximum insulin plasma levels and minimum glucose levels in blood are approximately 15 and 60 minutes respectively.

Rannsókn2

The mixtures from the comparative example and Example 1 were all given four or five dogs, with a Wright Dust Feed inhaler, with a constant scaling point of lUAg. The effects of each combination on plasma insulin levels and blood glucose levels were determined at different time points, and the results are shown in Figures 1 and 2. It was found that while the control compound containing no enhancement did not really induce any changes in plasma insulin levels, induced the insulin-induced mixture, an increase in plasma insulin levels of about 20 gU / ml over time to about 80 μυ / πύ 15 min. after inhalation of powder. Similarly, the comparative animals show, after inhalation of insulin, no insulin peak blood glucose, approximately 0.5 mmol / l, while the animals that induced insulin-induced insulin showed a transient decrease of about 1.7 mmol / 1, from about 4.0 mmol / l to about 25 nunol / l. Insulin associated with the egg, sodium caprate, was quickly absorbed into and purified from the cardiovascular cycle, with a corresponding transient decrease in the amount of glucose in the blood. Insulin with a burden (Iactose), but no enhancement was obviously only very small (p = 0.0002 for insulin / kaprat compared with insulin / lactose).

Rannsókn3

Blends from Examples 1-3 were all tested in different doses of two dogs. Blends were submitted with "Wright Dust Feed" inhaler. The level of insulin plasma and glucose in the blood was measured internally at different times. The results are shown in Figures 3-6 showing that insulin-related sodium caprate at different doses is rapidly absorbed and the maximum values ​​are reached after 20-30 minutes, followed by a corresponding decrease in the amount of glucose in the blood afterwards. The results also indicate that by inhalation of insulin powder, a plasmaferil that is more likely to be a natural physiological process than the curve obtained after a dermal dose of insulin can be obtained.

Dœmi4

Insulin and sodium caprate, 75:25: Mixture &amp; microwaved powder,

Lyfylated human insulin (53 g) was micronized in Airfílco Jet Mill (trademark Airfilco Process Plant Limited), with pressurized nitrogen (impulse pressure 7 bar, cell pressure 5 bar) to 2.4 micrometer mass extraction.

Sodium caprate (170 g) was micronized in Airfilco Jet Mill (TM), with nitrogen under pressure (impulse pressure 5 bar, cell pressure 3 bar), to 1.6 micrometer massametric diameter.

The micronized biologically-insulated human insulin (45 g) and sodium caprate (1456 g) were dry-blended according to the following procedure: Half of the insulin was placed in a shaker containing a blend solution of 4.4, respectively, divided by 1mm filter into two compartments with a metal ring in each compartment to assist in mixing and stirring. The sodium caprate was added to and eventually the rest of the insulin. The mixer was closed, turned 180 degrees and fitted to a motorized shaker. The machine was switched on and shaken at approximately

For two minutes until all insulin and sodium caprate were passed through the sift. The engine was switched off and the mixer dial turned 180 degrees, put back on the shaker and shake until all the powder passed through the filter. This procedure was repeated eight times or until the total mixing time had been about 20 minutes.

The resulting mixture was administered to five dogs by inhalation, used as a Wrigkt Dust Feed inhaler, at 1 U7kg doses and plasma levels of insulin were determined at different times after administration.

The results obtained were compared to plasma insulin levels obtained when the bioavailable insulin as microwaved as above to 2.4 micrometer MTP was given to five dogs in the same doses as in the plasma insulin level already obtained insulin and sodium caprate formulations in a ratio of 90:10 were administered five hundred in the same manner as in the same doses as above. In this case, the drug mixture was produced as follows: the semi-human human insulin was gel-filtered to reduce the relative content of sinks from 0.52% to 0.01% relative to insulin content. Insulin (45g) and sodium caprate (05g) were dissolved in water (232ml).

The solution was stirred until it became clear and the pH was adjusted to 7.0. The solution was concentrated by evaporation at 37 ° C in about two days. The solids cake obtained was pressed and filtered through a 05 mm filter and the powder obtained was micronized with particulate protection in particles with a 3.1 micrometer massametric diameter.

The results of this comparison are shown in Figure 9. The results show a somewhat improved bioavailability of insulin with the 90:10 blend and a decisive better bioavailability of insulin with the 75:25 blend of this invention, as compared to insulin alone (p = 0.0147 for the difference between between 75:25 and 100: 0).

Example 5

Vale exaggerate

The ability of all of the substances in Table ΙΠ to increase insulin absorption, thus affecting the amount of glucose in the blood, was investigated in rat model. Various forms of insulin were used:

manufactured with fusion technology either semi-synthetic or human. Each mixture was prepared as in Examples 1-3 above, wipe and work insulin / increase the mixture to produce inhaled powder.

Dust time was given rats by inhale and the follow-up was monitored by the amount of glucose in the blood. This amount was compared to the corresponding values ​​obtained in rats who received respiratory insufficiency.

Tafia ΠΙ

<td>Compound</td><td>Eykin insulin: lactose</td><td>effects</td>

<td>octylglucopyranoside</td><td>4: 4: 92</td><td>(+)</td>

<td>Natrfumúrsðdeoxýkólat</td><td>4: 4: 92</td><td>+</td>

<td>glycoside</td><td>4: 4: 92</td><td>+</td>

<td>Sodium</td><td>4: 4: 92</td><td>+</td>

<td>lysophospatidylcholine</td><td>4: 4: 92</td><td>+</td>

<td>dioctanoylphosphatidylcholine</td><td>2: 4: 94</td><td>(+)</td>

<td>Dídekanoýlfosfatidýlkólín</td><td>4: 4: 94</td><td></td>

<td>Sodium taurodihydrofusidate</td><td>2: 4: 94</td><td>+</td>

<td>Sodium caprylate</td><td>25: 75: 0</td><td>-</td>

<td>sodium</td><td>10: 90: 0</td><td>(+)</td>

<td>sodium</td><td>173: 823: 0</td><td>(+)</td>

<td>sodium</td><td>25: 75: 0</td><td>+</td>

<td>sodium</td><td>4: 4: 92</td><td>+</td>

<td>sodium laurate</td><td>25: 75: 0</td><td>L +)</td>

<td>Potassium oleate</td><td>4: 4: 92</td><td>+</td>

<td>Potassium caprate</td><td>27: 73: 0</td><td>+</td>

<td>Lysine caprate</td><td>35: 65: 0</td><td>+</td>

<td>Sodium myristate</td><td>30: 70: 0</td><td>+</td>

<td>Dimethyl-B-cyclodextrin</td><td>75: 25: 0</td><td>+</td>

+ effect, ie eykirran causes a significant reduction in the amount of glucose in the blood

- no very little effect (+) effect, but not as apparent as "+"

Example 6

Option enhancement

For the enhancer (sodium caprate) tested in the experiments, shown in Figures 5 and 6, the amount of labeling (mannitol, MW 360) appearing on the base and lateral side depends on the strength used by the enhancer, at least 16 mM of sodium caprate (Figure 7). This is correct even when adding an insulin / mannitol mixture (1: 3 sodium caprate insulin, by weight) (Figure 8). This concentration of sodium caprate (16 mM) also proved to promote insulin absorption through the simple cell layer. The amount of insulin that passed through the single layer doubled in the presence of 16 mM sodium caprate, compared to amount without any increase. It is expected that, in the case of subsidies of sodium caprate, the permeability of the cells will continue to increase;

This in vitro model of epithelial permeability can be used as a screening device to quickly test the utility of any desired enhancement in the methods of this invention.