JPH0348173B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a powder preparation of myeloperoxidase, which is prepared by adding albumin in an amount sufficient to stabilize myeloperoxidase. Myeloperoxidase (hereinafter abbreviated as MPO) was developed by Agner [Acta Physiol.Scand.] in 1941.
2, Suppl, 8 (1941)], it is an enzyme that was first isolated from human membranes, and is contained in large amounts together with lysozyme in cells of bone marrow origin, especially neutral polynucleated leukocytes and monocytes, and its content per neutrophil weight. is 5
reach %. This enzyme has a molecular weight of 120,000 to 150,000
It is a basic heme protein containing two atoms of iron per protein molecule, about the size of a Dalton, and belongs to oxidoreductases. The physiological function of MPO is thought to be that this enzyme kills or inactivates pathogenic microorganisms harmful to animals, such as bacteria, fungi, and viruses, in the presence of hydrogen peroxide and halogen ions. The usefulness of MPO as a medicine is known because a pharmaceutical composition containing MPO as a main ingredient shows dramatic therapeutic effects as a therapeutic agent for tuberculosis, such as isonicotinic acid hydrazide (INH)-resistant Mycobacterium tuberculosis infection. . This drug is usually packaged in ampoules or dispensed containers per unit dose as an injectable or topical composition, and is provided as a liquid, preferably a lyophilized powder preparation. It tends to be unstable when freeze-dried, and MPO activity decreases when freeze-dried or decreases over time. The present inventors conducted various studies to ensure the long-term storage stability of pharmaceutical compositions containing MPO as a main ingredient, and found that by adding albumin to MPO preparations, MPO activity during lyophilization was reduced. They also found that the decline in MPO activity over time during storage was suppressed, and moreover, the solubility of MPO preparations was increased, and the present invention was completed. The present invention is a powder formulation of MPO characterized in that albumin is added in an amount sufficient to stabilize MPO. Due to issues such as antigenicity, human-derived albumin is suitable for use in the present invention, and the degree of purification is sufficient as long as it has been purified for medical use, but albumin with a purity of 80% or more when analyzed by electrophoresis is suitable. preferable. The MPO used in the present invention is recovered from human leukocytes, cells of myeloid origin, particularly neutral polynucleated leukocytes, monocytes, etc. according to known production methods, and it is sufficient that the MPO is purified to a degree suitable for medical use. The MPO powder formulation according to the present invention is preferably a lyophilized product, and albumin may be added to the MPO solution before lyophilization and left in the powder without being removed, or immediately after lyophilization. Albumin may also be added. Additionally, monosaccharides, disaccharides, polysaccharides, sugar alcohols, amino acids, etc. may be added as stabilizers during freeze-drying, or they may be added after freeze-drying without any additives.
The amount of albumin added as a stabilizer is
Although it varies depending on the amount of MPO, the greater the amount added, the greater the stabilizing effect, and the amount added in powder form is generally 3 to 70 W/W%. Experiments 1 and 2 will explain the effects of albumin. The activity of MPO was measured by a modified method of B. Chance et al. using quaiacol. Experiment 1 Albumin was added as a stabilizer to a 100 unit/ml MPO solution before lyophilization, and the residual titer was examined immediately after lyophilization (A) and after storage at room temperature for 6 months (B). The results are shown in Table 1.

[Table] Experiment 2 A 100 unit/ml MPO solution was freeze-dried, immediately after which albumin was added as a stabilizer, and the remaining titer (%) was examined after storage at room temperature for 6 months.
The results are shown in Table 2.

[Table] These experiments showed that the addition of albumin was
It was found that it has a remarkable effect on the stability during freeze-drying and the stability over time of the freeze-dried product. Example 1 Albumin (1.0 W/V%) was added to a solution of myeloperoxidase at 100 units/ml and freeze-dried to obtain a dry powder myeloperoxidase preparation. Example 2 After 2.0 W/V% albumin was added to a solution of 150 units/ml of myeloperoxidase and lyophilized, 5% W/W albumin was further added to obtain a dry powder myeloperoxidase preparation. Example 3 20 W/W% albumin was added to a dry myeloperoxidase preparation to obtain a dry powder myeloperoxidase preparation.

Claims (1)

[Claims] 1. A powder preparation of myeloperoxidase, characterized in that it contains albumin in an amount sufficient to stabilize myeloperoxidase.