JPH0348173B2 - - Google Patents
Info
- Publication number
- JPH0348173B2 JPH0348173B2 JP56111017A JP11101781A JPH0348173B2 JP H0348173 B2 JPH0348173 B2 JP H0348173B2 JP 56111017 A JP56111017 A JP 56111017A JP 11101781 A JP11101781 A JP 11101781A JP H0348173 B2 JPH0348173 B2 JP H0348173B2
- Authority
- JP
- Japan
- Prior art keywords
- mpo
- albumin
- added
- myeloperoxidase
- freeze
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 102000009027 Albumins Human genes 0.000 claims description 17
- 108010088751 Albumins Proteins 0.000 claims description 17
- 102000003896 Myeloperoxidases Human genes 0.000 claims description 11
- 108090000235 Myeloperoxidases Proteins 0.000 claims description 11
- 239000000843 powder Substances 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 9
- 238000004108 freeze drying Methods 0.000 description 8
- 230000000694 effects Effects 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 4
- 239000003381 stabilizer Substances 0.000 description 4
- 238000003860 storage Methods 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 210000000265 leukocyte Anatomy 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- QRXWMOHMRWLFEY-UHFFFAOYSA-N isoniazide Chemical compound NNC(=O)C1=CC=NC=C1 QRXWMOHMRWLFEY-UHFFFAOYSA-N 0.000 description 2
- 239000002075 main ingredient Substances 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 201000008827 tuberculosis Diseases 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 102000008015 Hemeproteins Human genes 0.000 description 1
- 108010089792 Hemeproteins Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- -1 halogen ions Chemical class 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Description
本発明はミエロペルオキシダーゼにその安定化
に十分な量のアルブミンを添加してなるミエロペ
ルオキシダーゼの粉末製剤に関する。
ミエロペルオキシダーゼ(以下MPOと略記す
る)は1941年にAgner〔アクタ・フイジオロジ
カ・スカンジナビカ(Acta Physiol.Scand.),
2,Suppl,8(1941)〕により、ヒト膜よりはじ
めて分離された酵素で、骨髄起源細胞とくに中性
多核白血球および単球中にリゾチームとともに多
量に含まれており、好中球重量あたりの含量は5
%に達する。この酵素は分子量が120000〜150000
ダルトン程度、タンパク1分子あたり2原子の鉄
を含有する塩基性のヘムタンパク質であり、酸化
還元酵素に属する。MPOの生理機能は本酵素が
過酸化水素およびハロゲンイオンの共存下で細
菌、真菌、ウイルスなど動物に有害な病原微生物
を殺菌あるいは不活化するものと考えられてい
る。MPOの医薬としての有用性はMPOを主成分
とする医薬組成物が結核、たとえばイソニコチン
酸ヒドラジド(INH)耐性結核菌感染症の治療
剤として劇的な治療効果を示すことにより知られ
ている。
本剤は通常注射用または局所用組成物等として
単位投与量あたりのアンプルあるいは分注容器に
封入し、液状好ましくは凍結乾燥粉末製剤として
提供されるが、MPOを主成分とする医薬組成物
が凍結乾燥に対して不安定な傾向にあり、凍結乾
燥を行うとMPO活性が低下しあるいは経時的に
もMPO活性の減少が認められる。
本発明者らはMPOを主成分とする医薬組成物
の長期保存の安定性を確保するため種々の研究を
行い、その結果MPO製剤にアルブミンを添加す
ることにより、凍結乾燥時のMPO活性の低下な
らびに保存期間中の経時的なMPO活性の低下が
抑制され、しかもMPO製剤の溶解性を高めるこ
とを見いだし、本発明を完成した。
本発明はMPOを安定化するのに十分な量のア
ルブミンを添加したことを特徴とするMPOの粉
末製剤である。
本発明に用いるアルブミンは抗原性等の問題か
らヒト由来のものが適し、精製の度合は医療用と
して精製されたものであればよいが、電気泳動法
で分析して純度80%以上のものが好ましい。本発
明に用いるMPOはヒト白血球、骨髄性起源細胞
とくに中性多核白血球、単球等から公知の製法に
従つて回収され、純度は医療用に供しうる程度に
精製されておればよい。
本発明に係るMPO粉末製剤は好ましく凍結乾
燥品であり、アルブミンをMPO溶液の凍結乾燥
前に添加し、これを除去することなくそのまゝ粉
末中に存在させてもよく、凍結乾燥の直後にアル
ブミンを添加してもよい。また凍結乾燥時の安定
化剤として単糖類、二糖類、多糖類、糖アルコー
ル類、アミノ酸類等を添加してもよく、無添加で
凍結乾燥を行つた後にそれらを添加してもよい。
アルブミンの安定化剤としての添加量は製剤中の
MPO量によつて異なるが、その添加量が多いほ
ど安定化効果が大きく、一般に粉末状態での添加
量は3〜70W/W%である。
実験1、2によりアルブミンの効果を説明す
る。なおMPOの活性はクアヤコールを用いるビ
ー・チヤンス(B・Chance)らの改良法により
測定した。
実験1
100単位/mlのMPO溶液に凍結乾燥前に安定化
剤としてアルブミンを添加し、凍結乾燥直後(A)及
び室温にて6か月保存した際(B)の残存力価を調べ
た。その結果を第1表に示す。
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.
【表】
実験2
100単位/mlのMPO溶液を凍結乾燥し、その直
後に安定化剤としてアルブミンを添加し、室温に
て6か月保存した際の残存力価(%)を調べた。
その結果を第2表に示す。[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.
【表】
これらの実験によりアルブミンの添加がMPO
の凍結乾燥時の安定性及び凍結乾燥品の経時安定
性に顕著な効果を有することが判明した。
実施例 1
ミエロペルオキシダーゼの100単位/mlの溶液
にアルブミン1.0W/V%を添加し、凍結乾燥を
行なつて乾燥粉末のミエロペルオキシダーゼ製剤
を得た。
実施例 2
ミエロペルオキシダーゼの150単位/mlの溶液
にアルブミン2.0W/V%を添加して凍結乾燥を
行つた後、さらにアルブミン5W/W%を添加し
て乾燥粉末のミエロペルオキシダーゼ製剤を得
た。
実施例 3
ミエロペルオキシダーゼの乾燥製剤にアルブミ
ン20W/W%を添加し、乾燥粉末のミエロペルオ
キシダーゼ製剤を得た。[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)
分な量のアルブミンを添加したことを特徴とする
ミエロペルオキシダーゼの粉末製剤。1. A powder preparation of myeloperoxidase, characterized in that it contains albumin in an amount sufficient to stabilize myeloperoxidase.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP56111017A JPS5813521A (en) | 1981-07-16 | 1981-07-16 | Powdery pharmaceutical of myeloperoxidase |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP56111017A JPS5813521A (en) | 1981-07-16 | 1981-07-16 | Powdery pharmaceutical of myeloperoxidase |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5813521A JPS5813521A (en) | 1983-01-26 |
JPH0348173B2 true JPH0348173B2 (en) | 1991-07-23 |
Family
ID=14550279
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP56111017A Granted JPS5813521A (en) | 1981-07-16 | 1981-07-16 | Powdery pharmaceutical of myeloperoxidase |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5813521A (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108107210B (en) * | 2017-12-18 | 2019-01-04 | 广州市进德生物科技有限公司 | A kind of preparation method and frozen-dried protective liquid of myeloperoxidase freeze-drying calibration object |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS54147916A (en) * | 1978-05-12 | 1979-11-19 | Sumitomo Chem Co Ltd | Preparation of urokinase injection |
-
1981
- 1981-07-16 JP JP56111017A patent/JPS5813521A/en active Granted
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS54147916A (en) * | 1978-05-12 | 1979-11-19 | Sumitomo Chem Co Ltd | Preparation of urokinase injection |
Also Published As
Publication number | Publication date |
---|---|
JPS5813521A (en) | 1983-01-26 |
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