Patent Application
20080050438
The present invention is further described by reference to the following examples.
A suitable amount of distilled water for injection was poured into respective vials to give 1 ml of an injection comprising 0.1 ml of a drug substance in solution containing interferon-α (hereinafter referred to as “IFN-α bulk solution”, titer: 2×107 IU/ml), 5 mg of various amino acids and 1 mg of human serum albumin (HSA) per vial and subjected to lyophilization. Those samples were left to stand for three days under the conditions where the temperature was 40° C., relative humidity (RH) was 75% and the vials were left open (uncapped). Three days after, the titer of IFN-α was determined and the residual activity of INF-α was calculated by setting the IFN-α activity measured after drying to equal 100%.
Further, the same samples were evaluated for change in appearance after three days of standing under the conditions where the temperature was 40° C., RH was 75% and the vials were open. The results are shown in Table 1 below.
As is evident from the results summarized in Table 1, the products obtained by the present invention employing the hydrophobic amino acids having a Hydropathy Index of 3 or greater are remarkably superior in stability of IFN-α and/or change in appearance to the products in which other amino acids were employed, even when left in an excessively humid environment.
Deionized water was added to a mixture of 50 ml of an IFN-α bulk solution (titer: 2×107 IU/ml), 3500 mg of isoleucine and 700 mg of HSA, and then stirred thoroughly, to prepare 700 g of an IFN-α solution. To 700 g of this IFN-α solution was added 300 g of ethanol to give a weight ratio of water to ethanol of 7:3, and the solution to be spray-dried was produced.
Using a spray drier (Yamato Pulvis Basic Unit Model GB-21, manufactured by Yamato Science Co., Ltd.) under the conditions of air-supplying temperature of 130° C., spraying pressure of 2 kg/cm2 and spraying rate of 10 g/min, the above solution was spray-dried to produce dry particles.
Dry particles were produced in the similar manner as in (1) above with the exception that IFN-α was not employed.
The dry particles produced by the processes (1) and (2) above were each evaluated for aerodynamic average particle size (volume basis distribution), and the results are shown in Table 2 below. Aerodynamic average particle size was determined by dispersing the particles using an aerodisperser (Amherst Process Instruments, Inc.) and the measurement was conducted by using an aerosizer (Amherst Process Instruments, Inc.). Measuring conditions are as follows: air-stream shearing force: medium; sample particles supplying rate: medium; deagglomeration: normal; and vibration of dispersing pin: on.
Table 2 demonstrates that IFN-α does not affect the aerodynamic average particle size of the spray-dried products and the particle size distribution of the particles is dependent on the nature of amino acids employed.
To make a solution containing 0.5 wt % of each amino acid indicated in Table 3 and 0.1 wt % of HSA, suitable amount of deionized water was added to the solution and thoroughly stirred to prepare 700 g of an amino acid solution. To 700 g of this solution was added ethanol to give a weight ratio of water to ethanol of 7:3, and the solution to be spray-dried was produced.
Using a spray drier (Yamato Pulvis Basic Unit Model GB-21, manufactured by Yamato Science Co., Ltd.) under the conditions of air-supplying temperature of 130° C., spraying pressure of 2 kg/cm2 and spraying rate of 10 g/min, the above solution was spray-dried to produce the dry particles.
The dry particles produced by the above processes were each evaluated for moisture content (moisture content immediately after production and moisture content 24 hours after standing under the condition of RH 96%) and the average particle size distribution (volume basis distribution), and the results are summarized in Table 3 below.
Measurement of moisture content: the water contained in the dry particles were vaporized using Hiranuma auto moisture vaporizing instrument (LE-24S) and the moisture content was measured by using Hiranuma moisture microanalyzer (AQ-6).
Measurement of particle size: by using a laser diffraction scattering particle size distribution measuring equipment (LEM-24S, manufactured by Seishin Co., Ltd.), the particle size distribution of the dry particles (volume basis distribution) was determined. Measuring conditions were as follows: dispersing nozzle pressure: 5.0 kg/cm2; refractive index: 1.33.
The values shown in Table 3 are cumulative % under sieving. For example, “×50” indicates a particle size in which the particles of smaller sizes are accumulated to occupy 50% of the volume.
The dry particles produced using isoleucine, alanine or proline as the amino acid were evaluated for the particle size distribution by employing the above procedure and the graphs showing individual particle size distribution are represented in
As is evident from the results shown in Table 1 and
Dry particles were produced in the similar manner as in Example 2 with the exception that 300 g of ethanol was not added.
Dry particles were produced in the similar manner as in Example 2 with the exception that leucine, valine, leucyl-valine or isoleucyl-valyl-leucine was used in lieu of isoleucine.
Dry particles were produced in the similar manner as in Example 2 with the exception that an IFN-α bulk solution, isoleucine and HSA were employed in the amounts indicated in Table 4.
Dry particles were produced in the similar manner as in Example 4 with the exception that an IFN-α bulk solution, leucine and HSA were employed in the amounts indicated in Table 5.
A suitable amount of deionized water was added to a mixture of 50 ml of an IFN-α bulk solution (titer: 2×107 IU/ml), 3500 mg of isoleucine and 700 mg of HSA, and stirred thoroughly, to prepare 700 ml of an IFN-α solution. This solution was lyophilized, and the resultant lyophilized product was collected and milled using a jet-milling equipment to obtain dry particles.
Dry particles were produced in the similar manner as in Example 38 with the exception that an IFN-α bulk solution, isoleucine and HSA were employed in the amounts indicated in Table 6.
Dry particles were produced in the similar manner as in Example 38 by performing lyophilization with the exception that in lieu of isoleucine, 3500 mg of leucine was used.
Dry particles were produced in the similar manner as in Example 54 with the exception that an IFN-α bulk solution, leucine and HSA were employed in the amounts indicated in Table 7.
Dry particles were produced in the similar manner as in Example 2 with the exception that in lieu of the IFN-α bulk solution, 50 ml of an IFN-γ bulk solution (titer: 2×107 IU/ml) was used.
Dry particles were produced in the similar manner as in Example 2 with the exception that in lieu of the IFN-α bulk solution, 50 ml of a bulk solution containing interleukin-1β in which cysteine at position 71 was substituted with serine (described in European Patent Publication No. 237073A; titer: 1.2×108 IU/ml) was used.
Dry particles were produced in the similar manner as in Example 2 with the exception that in lieu of the IFN-α bulk solution, 50 ml of a bulk solution containing interleukin-1β in which asparagine at position 36 was substituted with aspartic acid and cysteine at position 141 was substituted with serine (described in European Patent Publication No. 237073A; titer: 1.3×108 IU/ml) was used.
Dry particles were produced in the similar manner as in Example 38 with the exception that in lieu of the IFN-α bulk solution, 50 ml of an IFN-γ bulk solution (titer: 2×107 IU/ml) was used.
Dry particles were produced in the similar manner as in Example 38 with the exception that in lieu of the IFN-α bulk solution, 50 ml of a bulk solution containing interleukin-1β in which cysteine at position 71 was substituted with serine (described in European Patent Publication No. 237073A; titer: 1.2×108 IU/ml) was used.
Dry particles were produced in the similar manner as in Example 38 with the exception that in lieu of the IFN-α bulk solution, 50 ml of a bulk solution containing interleukin-1β in which asparagine at position 36 was substituted with aspartic acid and cysteine at position 141 was substituted with serine (described in European Patent Publication No. 237073A; titer: 1.2×108 IU/ml) was used.
Dry particles were produced in the similar manner as in Example 2 with the exception that the IFN-α bulk solution, hydrophobic stabilizers (leucine and valine) and other stabilizers (glycine, sucrose or mannitol) were employed in the amounts indicated in Table 8.
Dry particles were produced in the similar manner as in Example 38 with the exception that the IFN-α bulk solution, hydrophobic stabilizers (leucine and valine) and other stabilizers (glycine, sucrose or mannitol) were employed in the amounts indicated in Table 9.
| Number | Date | Country | Kind |
|---|---|---|---|
| 336714/1995 | Dec 1995 | JP | national |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 10143815 | May 2002 | US |
| Child | 11516728 | US | |
| Parent | 09091676 | Jun 1998 | US |
| Child | 10143815 | US |
