Abstract
The invention relates to the use of an aerosol-generating device for generating an aerosol of a composition, for administering the composition by inhalation, wherein the composition comprises at least one antisense-molecule. The aerosol-generating device has a membrane having a perforation, wherein the composition for generating the aerosol is brought into contact with the membrane, and the membrane of the aerosol-generating device are caused to vibrate. The perforation is formed of at least one group of passage openings, wherein each passage opening of a group has a diameter of at least 3 μm and at most 5 μm at the narrowest point of the passage opening, and wherein the at least one group of passage openings comprises at maximum 500 passage openings per mm 2 of the membrane.
Claims
1. A method for generating an aerosol of a composition, for administering the composition by inhalation, where the composition comprises at least one antisense molecule, where the aerosolizer has a membrane with a perforation, where the composition, for generating the aerosol, is contacted with the membrane, and the membrane of the aerosolizer is set in vibration, where the perforation is formed by at least one group of passages, and where each passage of a group has a diameter at its narrowest point of at least 3 μm and at most 5 μm and where the at least one group of passages comprises at maximum 500 passages per mm.sup.2 of the membrane.
2. The method of claim 1, characterized in that the membrane has a piezoelectric element which sets the membrane in vibration.
3. The method of claim 1, characterized in that the passages of the membrane are cylindrical.
4. The method of claim 1, characterized in that the passages of the perforation are conical, with the diameter of each conical passage decreasing in the direction of the region of the perforation at which the aerosol emerges from the perforation.
5. The method of claim 1, characterized in that the perforation of the membrane has at least two groups of passages.
6. The method of claim 1, characterized in that the membrane has a diameter of 6 mm to 8 mm.
7. The method of claim 1, characterized in that the membrane is produced from a material which comprises at least one metal, the metal being more particularly an element from the group of stainless steel, nickel, palladium, cobalt, rust-free steel and/or being an alloy of at least two of the elements.
8. The method of claim 1, characterized in that the composition has an antisense molecule concentration of below 75 mg/ml.
9. The method of claim 8, characterized in that the composition has an antisense molecule concentration of 20 mg/ml to 50 mg/ml.
10. The method of claim 1, characterized in that the composition has a viscosity which is lower than 3.5 mPa.Math.s.
11. The method of claim 1, characterized in that the group of passages comprises at maximum 350 passages per mm.sup.2 of the membrane.
12. The method of claim 1, characterized in that the group of passages comprises at maximum 200 passages per mm.sup.2 of the membrane.
13. The method of claim 1, characterized in that the group of passages comprises at maximum 50 passages per mm.sup.2 of the membrane.
14. The method of claim 1, characterized in that the composition comprises at least one antisense molecule which is selected from the group encompassing DNAzymes, siRNAs, asDNAs or Ribozymes.
15. The method of claim 14, characterized in that the composition comprises at least one antisense molecule which specifically downregulates the expression of GATA-3, or comprises at least one antisense molecule which specifically downregulates the expression of Tbet, where the antisense molecule is selected from a group encompassing the DNAzymes hdg1 to hdg70 or from a group encompassing the DNAzymes td1 to td78.
16. The method of claim 15, characterized in that the at least one antisense molecule is a DNAzyme which has the hdg40 sequence (GTGGATGGAggctagctacaacgaGTCTTGGAG).
Description
FIGURES
[0048] FIG. 1 shows a graph of the viscosity of the antisense molecule-containing composition as a function of its concentration.
[0049] FIG. 2 shows results of viscosity measurements on the antisense molecule-containing composition as a function of its concentration.
[0050] FIG. 3 shows measurement results of an FAT (factory acceptance test) of compositions with different antisense molecule concentrations of the antisense molecule-containing composition.
[0051] FIG. 4 shows measurement results of an FAT of the antisense molecule-containing composition with antisense molecule concentrations of 20 mg/ml and 50 mg/ml.
[0052] FIG. 5 shows measurement results of the antisense molecule-containing composition which were by various aerosolizers with membranes having different perforations.
[0053] FIG. 1 shows a graph of the viscosity of an antisense molecule-containing composition as a function of its concentration. In the example shown, the antisense molecule is a DNAzyme. This DNAzyme can specifically downregulate the expression of GATA-3 and/or the expression of Tbet. In a preferred version, antisense molecules in particular from a group encompassing the DNAzymes hdg1 to hdg70 or from a group encompassing the DNAzymes td1 to td78 are provided. Preference is given to using the DNAzyme hgd40 with the sequence GTGGATGGAggctagctacaacgaGTCTTGGAG. Hdg40 specifically inhibits the expression of GATA-3. FIG. 1 shows a graph, illustratively, of the viscosity of an hgd40-containing composition. Along the Y axis of the graph from FIG. 1, the viscosity of the composition is plotted in mPa.Math.s; the X axis indicates the DNAzyme concentration of the composition in mg/ml. From the plot it is apparent that the viscosity of the DNAzyme-containing composition undergoes a substantially exponential increase. A sharp increase in the viscosity can be observed in particular at a concentration of 50 mg/ml and higher. A viscosity of 3.5 mPa.Math.s is observable even at a concentration of about 75 mg/ml.
[0054] FIG. 2 shows the results of a viscosity measurement on an antisense molecule-containing composition as a function of its concentration, with the antisense molecules studied being DNAzymes. The results set out as a table demonstrate the exponential increase in the viscosity of an antisense molecule-containing composition having an antisense molecule concentration of between 50 mg/ml and 100 mg/ml. The correlation shown here between nucleic acid concentration and viscosity is not so pronounced for different, double-stranded DNA molecules; the viscosity of a double-stranded DNA-containing solution typically sees a substantially linear increase with increasing DNA concentration. At a viscosity of above 3.5 mPa.Math.s, the pressure which an aerosolizer would have to achieve is too high. This may result in the passages or holes in the membrane becoming clogged or fouled. This risk is significantly minimized by a composition whose antisense molecule concentration is lower than 75 mg/ml, because the viscosity of such an antisense molecule-containing composition is below 3.5 mPa.Math.s.
[0055] FIG. 3 shows in table form measurement results of an FAT (factory acceptance test) of compositions having different antisense molecule concentrations, with the antisense molecules being DNAzymes, specifically the DNAzyme hgd40. The purpose of an FAT is to test the usability of a product. The table shows that a composition having an antisense molecule concentration of above 75 mg/ml can no longer be misted by the aerosolizer from the test (cf. line 3 of the table from FIG. 3). If a composition having an antisense molecule concentration of 75 mg/ml is subjected to sterile filtration in advance of the test, the antisense molecule concentration is lowered to below 75 mg/ml. A composition having a DNAzyme concentration of this kind can be misted using the aerosolizer employed in the test (cf. lines 5 and 6 of the table from FIG. 3). The FAT reports the viscosity (dP/ml) in percent. In order to pass the FAT carried out here, the viscosity had to be below 5%. The abbreviation “WFI” in the second and fourth lines of the table from FIG. 3 stands for “Water for Injection” and represents the test control—accordingly, the “WFI” controls correspond to a viscosity of 1 dP/ml.
[0056] FIG. 4 shows measurement results of an FAT on a composition having antisense molecule concentrations of 20 mg/ml and 50 mg/ml, respectively. The antisense molecules studied here comprise, in particular, DNAzymes. The results show that such compositions pass the FAT and can be misted by means of the use according to the invention (cf. “passed” in lines 3, 4, 6 and 7 of the table from FIG. 4).
[0057] The measurement results from FIGS. 3 and 4 therefore show that an antisense molecule-containing composition, and more particularly a DNAzyme-containing composition having a DNAzyme concentration of below 75 mg/ml, can be used as an aerosol by means of an aerosolizer in accordance with the features presently described. Hence the use according to the invention is ideally suited to the treatment of a patient suffering from a respiratory tract disease associated with chronic inflammation. Preference here is given to using antisense molecule or DNAzyme concentrations of 20 mg/ml to 50 mg/ml.
[0058] FIG. 5 shows measurement results on some of the aerosolizers tested. The measurement results show that aerosolizers having the features of the invention are ideally suited to generating an aerosol of an antisense molecule-containing composition (FIG. 5, columns B-D). In the example shown in FIG. 5, the antisense molecules are DNAzymes. In the case of the devices from columns B-D, the aerosol is formed by bringing the composition into contact with the aerosolizer membrane which has been set in vibration. In the invention this membrane has a perforation which is formed by at least one group of passages, the passages at their narrowest point having a diameter which is between 3 μm and 5 μm (FIG. 5, cells B5, C5 and D5), with the group of passages comprising at maximum 500 passages per mm.sup.2 of the membrane (FIG. 5, cells B6, C6 and D6). In this case, droplets or particles having a size of around 4 μm are produced (FIG. 5, cells B11-B13, C14, D12 and D14), this being regarded as ideal for the generation of an aerosol of an antisense molecule-containing composition for the treatment of a patient suffering from a respiratory tract disease associated with chronic inflammation. The vibration of the membrane boosts the formation of droplets having the preferred size.
[0059] A device in which the passages at their narrowest point have a diameter of less than 3 μm (FIG. 5, cell 5A) or whose membrane has more than 500 passages per mm.sup.2 (FIG. 5, cell 6A) results in an aerosol with droplets having a size of more than 5 μm VMD MV (FIGS. 12A, 13A and 14A). Droplets of this size have the drawback that because of impactation forces, they do not reach their corresponding site of action, or at least do so only to a greatly reduced extent. Accordingly, the antisense molecules and in particular the DNAzymes would not be able to develop their therapeutic effect at the site at which the chronic inflammation is present. One of the reasons for this is the hole density: with a hole density of above 500 passages per mm.sup.2 of the membrane, it is possible for the resultant droplets or particles to undergo fusion, aggregation or combination to form larger particles directly after emergence from the passages.
