JPS6326120B2 - - Google Patents
Info
- Publication number
- JPS6326120B2 JPS6326120B2 JP54009939A JP993979A JPS6326120B2 JP S6326120 B2 JPS6326120 B2 JP S6326120B2 JP 54009939 A JP54009939 A JP 54009939A JP 993979 A JP993979 A JP 993979A JP S6326120 B2 JPS6326120 B2 JP S6326120B2
- Authority
- JP
- Japan
- Prior art keywords
- interferon
- concentration
- added
- freeze
- activity
- 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
Links
- 102000014150 Interferons Human genes 0.000 claims description 33
- 108010050904 Interferons Proteins 0.000 claims description 33
- 229940079322 interferon Drugs 0.000 claims description 32
- -1 polyethylene Polymers 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 11
- 229920003171 Poly (ethylene oxide) Polymers 0.000 claims description 4
- 239000004698 Polyethylene Substances 0.000 claims description 4
- 239000002736 nonionic surfactant Substances 0.000 claims description 4
- 229920000573 polyethylene Polymers 0.000 claims description 4
- 229920001451 polypropylene glycol Polymers 0.000 claims description 4
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 claims description 4
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims description 3
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 3
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 3
- 239000002202 Polyethylene glycol Substances 0.000 claims description 3
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims description 3
- 239000003242 anti bacterial agent Substances 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 3
- 229920001223 polyethylene glycol Polymers 0.000 claims description 3
- UEUXEKPTXMALOB-UHFFFAOYSA-J tetrasodium;2-[2-[bis(carboxylatomethyl)amino]ethyl-(carboxylatomethyl)amino]acetate Chemical compound [Na+].[Na+].[Na+].[Na+].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O UEUXEKPTXMALOB-UHFFFAOYSA-J 0.000 claims description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 3
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 3
- 229930182555 Penicillin Natural products 0.000 claims description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 2
- 229960000318 kanamycin Drugs 0.000 claims description 2
- 229930027917 kanamycin Natural products 0.000 claims description 2
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 claims description 2
- 229930182823 kanamycin A Natural products 0.000 claims description 2
- 229940049954 penicillin Drugs 0.000 claims description 2
- 229960005322 streptomycin Drugs 0.000 claims description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims 3
- 230000003115 biocidal effect Effects 0.000 claims 2
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 claims 2
- BSYNRYMUTXBXSQ-FOQJRBATSA-N 59096-14-9 Chemical compound CC(=O)OC1=CC=CC=C1[14C](O)=O BSYNRYMUTXBXSQ-FOQJRBATSA-N 0.000 claims 1
- 239000005635 Caprylic acid (CAS 124-07-2) Substances 0.000 claims 1
- 125000004432 carbon atom Chemical group C* 0.000 claims 1
- 229960004106 citric acid Drugs 0.000 claims 1
- 229960002446 octanoic acid Drugs 0.000 claims 1
- 150000007524 organic acids Chemical class 0.000 claims 1
- 150000003839 salts Chemical class 0.000 claims 1
- 230000006641 stabilisation Effects 0.000 claims 1
- 238000011105 stabilization Methods 0.000 claims 1
- 230000000694 effects Effects 0.000 description 20
- 238000004108 freeze drying Methods 0.000 description 12
- 239000000243 solution Substances 0.000 description 8
- 230000000087 stabilizing effect Effects 0.000 description 7
- 239000003814 drug Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 210000004698 lymphocyte Anatomy 0.000 description 3
- 239000003381 stabilizer Substances 0.000 description 3
- 241000282412 Homo Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 230000005856 abnormality Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- HFWWEMPLBCKNNM-UHFFFAOYSA-N n-[bis(hydroxyamino)methyl]hydroxylamine Chemical compound ONC(NO)NO HFWWEMPLBCKNNM-UHFFFAOYSA-N 0.000 description 2
- 238000005185 salting out Methods 0.000 description 2
- 229960005480 sodium caprylate Drugs 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
- BYKRNSHANADUFY-UHFFFAOYSA-M sodium octanoate Chemical compound [Na+].CCCCCCCC([O-])=O BYKRNSHANADUFY-UHFFFAOYSA-M 0.000 description 2
- JZLOKWGVGHYBKD-UHFFFAOYSA-M sodium;2-acetyloxybenzoate Chemical compound [Na+].CC(=O)OC1=CC=CC=C1C([O-])=O JZLOKWGVGHYBKD-UHFFFAOYSA-M 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000004094 surface-active agent Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 241000711975 Vesicular stomatitis virus Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 210000001691 amnion Anatomy 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- HQPMKSGTIOYHJT-UHFFFAOYSA-N ethane-1,2-diol;propane-1,2-diol Chemical compound OCCO.CC(O)CO HQPMKSGTIOYHJT-UHFFFAOYSA-N 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 229940047124 interferons Drugs 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920001993 poloxamer 188 Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Description
インターフエロンは、ウイルスあるいはその他
の物質の刺激によりヒトを含む動物細胞から産生
されるある種の糖蛋白質である。
このものは、ウイルスや細菌又は原虫の増殖を
阻止するが、その作用は動物種に特異的であるこ
とから医薬としてヒトに用いる場合は、ヒトの細
胞から産生されたインターフエロンを得る必要が
ある。
ヒトのインターフエロンを大量に得るために
は、ヒトリンパ球、ヒト繊維芽細胞又はヒト株化
リンパ芽球等を大量に集め、これらの細胞に適当
な刺激を与えてインターフエロンを産生せしめ、
更にこの産生されたインターフエロンをアフイニ
テイークロマトグラフイーやイオン交換クロマト
グラフイー又は塩析法等で精製することが必要で
ある。
しかし、ヒトインターフエロンは活性の安定性
が乏しく精製するに従い活性の損失が著しく特に
凍結乾燥時には激減することが多い。それがため
にヒトインターフエロンを大量に精製し医薬とし
て用いるまでには多大の労力と時間と多量の原料
を要していた。
本発明者らは、ヒトインターフエロンの精製研
究とともにこのものの活性を安定化させる方法に
ついても長年検討を続け、トリスヒドロキシメチ
ルアミノメタン、ポリオキシエチレン・ポリオキ
シプロピレン共重合体、ポリエチレングリコール
の様なポリエチレン系非イオン界面活性剤、抗生
物質、有機酸塩、エチレンジアミン４酢酸ナトリ
ウムの様なキレート剤および芳香族アミノ酸等を
単独か又はこれらの複数をインターフエロンを含
む水溶液に添加することにより製造工程中特に凍
結乾燥時のインターフエロンの活性が著しく安定
化されることを見出し、本発明を完成せしめた。
トリスヒドロキシメチルアミノメタンは、溶液
とするとアルカリ性となるが、このものを終濃度
が0.001M以上好ましくは0.001M〜0.10Mより好
ましくは0.01M〜0.05Mになるように精製インタ
ーフエロン溶液に加え、酢酸又は塩酸でPHを5.0
〜8.0に修正し凍結乾燥することにより、インタ
ーフエロン活性は著しく安定化される。この時の
PHは特に限定する必要はないが生理的なPHである
方が、乾燥品を溶解し、生体内に投与する場合に
都合が良い。
ポリエチレン系の非イオン界面活性剤の分子量
が、2000から20000のものは界面活性剤として広
く利用されているが、本発明者らはこのものがイ
ンターフエロン活性の安定化作用のあることを偶
然見出した。またこのものの分子量とインターフ
エロン活性の安定化効果との関係は明瞭ではなく
いずれの分子量のものも安定化作用を示すが好ま
しくは凍結乾燥が容易なことから分子量が5000〜
10000のものを用いる。用いる濃度は、終濃度が
0.001w／ｖ％以上好ましくは0.001w／ｖ％〜
0.5w／ｖ％になるようにインターフエロン溶液
に加える。
ポリエチレン系の非イオン界面活性剤の例示と
しては、
ポリエチレングリコールの場合もポリオキシエ
チレン・ポリオキシプロピレン共重合体の場合も
同様の方法で加えることにより著しい安定化効果
が示される。
その他インターフエロン活性の凍結乾燥時の安
定化剤としては、
カナマイシン、ペニシリン、ストレプトマイシ
ンなどの抗生物質、カプリル酸ナトリウム、クエ
ン酸ナトリウム、アセチルサリチル酸ナトリウム
等の有機酸塩、フエニルアラニン、チロシンおよ
びトリプトフアンなどの芳香族アミノ酸等々は対
照群と比べインターフエロン活性を著しく安定化
させることが判明した。
これらの用いる濃度は、0.0001w／ｖ％以上好
ましくは0.0001w／ｖ％〜0.2w／ｖ％で十分な安
定化効果がみられるがより好ましくはカプリル酸
ナトリウム、クエン酸ナトリウムおよびアセチル
サリチル酸ナトリウムでは0.001w／ｖ％以上好
ましくは0.001％〜0.1％、フエニルアラニン、チ
ロシンおよびトリプトフアンでは0.01w／ｖ％以
上好ましくは0.01w／ｖ％〜0.5％が経済的な有効
濃度である。
以上の安定化剤の添加量の上限は、医薬として
の含有するに際し、許容される添加量によつて特
定されるものであつて、添加による副作用が認め
られないかぎり特に限定されない。
かくして凍結乾燥されたインターフエロン製剤
は、そのまま医薬に利用されうる。
本発明におけるインターフエロン活性の測定
は、ヴエシクラー ストマテイテイス ウイルス
（Vesicular stomatitis virus）とヒト羊膜由来の
FL細胞を用い、50％プラツク減少法により測定
した。インターフエロン力価は、被検試料と同時
に測定したスタンダード・インターフエロンの力
価より国際単位（IU）に換算した。（最近医薬29
(4)660、1974）
実施例 １
ヒト静脈血由来のリンパ球より硫安塩析および
イオン交換吸着法で部分精製した比活性５×105
国際単位／O.D.280n.m.のインターフエロン溶液に
0.01Mのトリスヒドロキシアミノメタンを添加し
Ｎ−HClでPHを7.2±0.1に修正し除菌過を行つ
た後、凍結乾燥を行つた。凍結乾燥品を0.10Mリ
ン酸緩衝液PH7.2に溶解しインターフエロンの残
存活性を測定した。
対照として0.5％塩化ナトリウム溶液PH7.2±0.1
で同時に凍結乾燥を行いその残存活性を測定し
た。
その結果、凍結乾燥前のインターフエロン活性
を100％とすると対照品では、35±20％残存活性
を示したのに比べ本発明の方法で凍結乾燥したも
のでは100±10％の残存活性を示した。
また、本品の溶解液を用いラツトに30×104単
位を腹腔内投与し、７日間の観察を続けたが何ら
異常は認められず安全性の高いことが判つた。
実施例 ２
実施例１と同様の方法で得られたインターフエ
ロン溶液にプルロニツクF68（ポリオキシエチレ
ン・ポリオキシプロピレン共重合体）（平均分子
量8350）を0.01％の濃度に加え、除菌過を行つ
た後凍結乾燥を行つた。
インターフエロン活性の残存率は、凍結乾燥前
を100％として120％であり凍結乾燥時の失活は完
全に抑えられた。また、この凍結乾燥後のインタ
ーフエロンを生理食塩水に溶解し、20±２ｇのマ
ウス３匹に５×104国際単位／匹を投与して７日
間の観察を続けたが観察期間中何ら異常は認めら
れなかつた。
実施例 ３
ナマルバ株のリンパ球によつて誘発産生された
インターフエロン（特願昭53−16341）をSP−セ
フアデツクス（フアーマシヤ製）およびセフアデ
ツクスＧ−100を用いて精製（特願昭52−156019）
されたヒトインターフエロン溶液を用いる以外は
実施例１を繰り返えした。
その結果、活性残存率及び安全性において実施
例１と同様の結果を得た。
実験例
実施例１において凍結乾燥時の安定化剤とし
て、トリスヒドロキシアミノメタン0.01M添加の
代りに他の化合物を種々変えてその安定化効果を
比較検討した。試料は凍結乾燥前のインターフエ
ロン活性を100％とし、凍結乾燥後の残存活性率
を測定した。なお添加量は凍結乾燥前の水溶液中
での濃度として表わした。
結果を第１表、第２表及び第３表に示す。
Interferons are a type of glycoprotein produced by animal cells, including humans, upon stimulation by viruses or other substances. This substance inhibits the growth of viruses, bacteria, and protozoa, but its action is specific to animal species, so if it is to be used as a medicine in humans, it is necessary to obtain interferon produced from human cells. . In order to obtain a large amount of human interferon, a large number of human lymphocytes, human fibroblasts, human lymphoblast cell lines, etc. are collected, and appropriate stimulation is given to these cells to produce interferon.
Furthermore, it is necessary to purify the produced interferon by affinity chromatography, ion exchange chromatography, salting out method, or the like. However, human interferon has poor activity stability, and as it is purified, its activity is often significantly reduced, especially during freeze-drying. Therefore, it took a great deal of effort, time, and large amounts of raw materials to purify human interferon in large quantities and use it as a medicine. The present inventors have continued to research purification of human interferon and methods for stabilizing its activity for many years. During the manufacturing process, polyethylene nonionic surfactants, antibiotics, organic acid salts, chelating agents such as sodium ethylenediaminetetraacetate, aromatic amino acids, etc., singly or in combination, are added to the aqueous solution containing interferon. In particular, we have found that the activity of interferon during freeze-drying is significantly stabilized, and have completed the present invention. Trishydroxymethylaminomethane becomes alkaline when made into a solution, but it is added to the purified interferon solution so that the final concentration is 0.001M or more, preferably 0.001M to 0.10M, more preferably 0.01M to 0.05M, Adjust pH to 5.0 with acetic acid or hydrochloric acid
By fixing to ~8.0 and lyophilizing, interferon activity is significantly stabilized. at this time
Although the pH does not need to be particularly limited, a physiological pH is more convenient when dissolving a dried product and administering it to a living body. Polyethylene-based nonionic surfactants with a molecular weight of 2,000 to 20,000 are widely used as surfactants, but the present inventors accidentally discovered that this surfactant has a stabilizing effect on interferon activity. Ta. Furthermore, the relationship between the molecular weight and the stabilizing effect on interferon activity is not clear, and although any molecular weight shows a stabilizing effect, it is preferable to use a molecular weight of 5,000 to 5,000 because it is easy to freeze-dry.
Use 10000. The final concentration used is
0.001w/v% or more preferably 0.001w/v%~
Add to interferon solution at 0.5w/v%. As examples of polyethylene-based nonionic surfactants, both polyethylene glycol and polyoxyethylene/polyoxypropylene copolymer exhibit remarkable stabilizing effects when added in the same manner. Other stabilizers for interferon activity during freeze-drying include antibiotics such as kanamycin, penicillin, and streptomycin, organic acid salts such as sodium caprylate, sodium citrate, and sodium acetylsalicylate, phenylalanine, tyrosine, and tryptophan. aromatic amino acids, etc. were found to significantly stabilize interferon activity compared to the control group. A sufficient stabilizing effect can be seen at a concentration of 0.0001w/v% or more, preferably from 0.0001w/v% to 0.2w/v%, and more preferably sodium caprylate, sodium citrate, and sodium acetylsalicylate. The economically effective concentration is 0.001 w/v % or more, preferably 0.001% to 0.1%, and for phenylalanine, tyrosine and tryptophan, 0.01 w/v % or more, preferably 0.01 w/v % to 0.5%. The upper limit of the amount of the above-mentioned stabilizer added is determined by the allowable amount added when included as a medicine, and is not particularly limited as long as no side effects are observed due to the addition. The thus freeze-dried interferon preparation can be used as is in medicine. The measurement of interferon activity in the present invention is performed using Vesicular stomatitis virus and human amnion-derived interferon.
It was measured by the 50% plaque reduction method using FL cells. The interferon titer was converted into international units (IU) from the standard interferon titer measured at the same time as the test sample. (recently pharmaceutical 29
(4)660, 1974) Example 1 Specific activity 5×10 5 partially purified from lymphocytes derived from human venous blood by ammonium sulfate salting out and ion exchange adsorption method.
Interferon solution with international units/OD280 nm
After adding 0.01M trishydroxyaminomethane and correcting the pH to 7.2±0.1 with N-HCl for sterilization, freeze-drying was performed. The lyophilized product was dissolved in 0.10M phosphate buffer PH7.2, and the residual activity of interferon was measured. 0.5% sodium chloride solution PH7.2±0.1 as control
At the same time, it was freeze-dried and its residual activity was measured. As a result, when the interferon activity before freeze-drying is taken as 100%, the control product showed a residual activity of 35±20%, while the product freeze-dried by the method of the present invention showed a residual activity of 100±10%. Ta. In addition, 30 x 10 4 units of the solution of this product was intraperitoneally administered to rats, and observation was continued for 7 days, but no abnormalities were observed and it was found to be highly safe. Example 2 Pluronic F68 (polyoxyethylene/polyoxypropylene copolymer) (average molecular weight 8350) was added to an interferon solution obtained in the same manner as in Example 1 at a concentration of 0.01%, and sterilization was carried out. After drying, freeze-drying was performed. The residual rate of interferon activity was 120%, taking the value before freeze-drying as 100%, and deactivation during freeze-drying was completely suppressed. In addition, this freeze-dried interferon was dissolved in physiological saline, and 5 x 10 4 international units/mouse was administered to three mice weighing 20 ± 2 g, and observation was continued for 7 days, but no abnormalities were observed during the observation period. was not recognized. Example 3 Interferon (patent application 16341/1989) induced and produced by lymphocytes of the Namalva strain was purified using SP-Sephadex (manufactured by Pharmacia) and Cephadex G-100 (Japanese patent application 156019/1982)
Example 1 was repeated except using the human interferon solution obtained. As a result, results similar to those of Example 1 were obtained in terms of residual activity and safety. Experimental Example In Example 1, instead of adding 0.01 M of trishydroxyaminomethane as a stabilizer during freeze-drying, various other compounds were used and their stabilizing effects were compared and studied. The interferon activity of the sample before freeze-drying was set to 100%, and the residual activity rate after freeze-drying was measured. The amount added was expressed as the concentration in the aqueous solution before freeze-drying. The results are shown in Tables 1, 2 and 3.
【表】【table】
【表】【table】
【表】【table】
【表】【table】
Claims (1)
オキシエチレン・ポリオキシプロピレン共重合
体、ポリエチレングリコールから選ばれたポリエ
チレン系非イオン界面活性剤、カナマイシン、ス
トレプトマイシン、ペニシリンから選ばれた抗生
物質、カプリル酸、クエン酸、アセチルサリチル
酸から選ばれた炭素数４〜10の有機酸の塩、エチ
レンジアミン４酢酸ナトリウム、および芳香族ア
ミノ酸のいずれかをインターフエロンを含有する
水溶液に添加し凍結乾燥することを特徴とするイ
ンターフエロンの安定化法。 ２ トリスヒドロキシメチルアミノメタンを
0.001M濃度以上に加えPHを5.0〜8.0にすることを
特徴とする特許請求の範囲第１項に記載の方法。 ３ ポリオキシエチレン・ポリオキシプロピレン
共重合体の平均分子量が2000から20000のものを
0.001w／ｖ％〜0.5w／ｖ％濃度以上に加え、PH
を3.0〜8.0にすることを特徴とする特許請求の範
囲第１項に記載の方法。 ４ ポリエチレングリコールの分子量が、2000〜
20000のものを0.001w／ｖ％濃度以上に加えるこ
とを特徴とする特許請求の範囲第１項に記載の方
法。 ５ 抗生物質を0.0001w／ｖ％濃度以上に加える
ことを特徴とする特許請求の範囲第１項に記載の
方法。 ６ 有機酸塩を0.0001w／ｖ％濃度以上に加え、
PH7.0〜8.0とすることを特徴とする特許請求の範
囲第１項に記載の方法。 ７ エチレンジアミン４酢酸ナトリウムを
0.001M濃度以上に加えることを特徴とする特許
請求の範囲第１項に記載の方法。 ８ 芳香族アミノ酸が、フエニルアラニン、チロ
シンおよびトリプトフアンであり、これを濃度が
0.01w／ｖ％以上になるように加えることを特徴
とする特許請求の範囲第１項に記載の方法。[Claims] 1. A polyethylene nonionic surfactant selected from trishydroxymethylaminomethane, polyoxyethylene/polyoxypropylene copolymer, and polyethylene glycol, an antibiotic selected from kanamycin, streptomycin, and penicillin, A salt of an organic acid having 4 to 10 carbon atoms selected from caprylic acid, citric acid, and acetylsalicylic acid, sodium ethylenediaminetetraacetate, and an aromatic amino acid are added to an aqueous solution containing interferon and freeze-dried. Characteristic stabilization method for interferon. 2 Trishydroxymethylaminomethane
The method according to claim 1, characterized in that the concentration is 0.001M or more and the pH is adjusted to 5.0 to 8.0. 3 Polyoxyethylene/polyoxypropylene copolymer with an average molecular weight of 2,000 to 20,000.
In addition to the concentration of 0.001w/v% to 0.5w/v% or more, PH
3.0 to 8.0. 4 The molecular weight of polyethylene glycol is 2000~
20,000 at a concentration of 0.001 w/v% or more. 5. The method according to claim 1, characterized in that the antibiotic is added at a concentration of 0.0001 w/v% or more. 6 Add organic acid salt to a concentration of 0.0001w/v% or more,
The method according to claim 1, characterized in that the pH is 7.0 to 8.0. 7 Sodium ethylenediaminetetraacetate
2. The method according to claim 1, characterized in that it is added at a concentration of 0.001M or more. 8 Aromatic amino acids are phenylalanine, tyrosine, and tryptophan, which are
The method according to claim 1, characterized in that the amount is added to be 0.01 w/v% or more.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP993979A JPS55102519A (en) | 1979-01-31 | 1979-01-31 | Stabilization of interferon |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP993979A JPS55102519A (en) | 1979-01-31 | 1979-01-31 | Stabilization of interferon |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS55102519A JPS55102519A (en) | 1980-08-05 |
| JPS6326120B2 true JPS6326120B2 (en) | 1988-05-27 |
Family
ID=11733977
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP993979A Granted JPS55102519A (en) | 1979-01-31 | 1979-01-31 | Stabilization of interferon |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS55102519A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04200105A (en) * | 1990-11-29 | 1992-07-21 | Nec Corp | Travelling wave tube power amplifier |
Families Citing this family (22)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA1190148A (en) * | 1981-10-13 | 1985-07-09 | Samuel S. Asculai | Interferon-containing compositions |
| US4507281A (en) * | 1981-10-13 | 1985-03-26 | Exovir, Inc. | Interferon-containing compositions |
| JPS5892621A (en) * | 1981-11-28 | 1983-06-02 | Sunstar Inc | Stable pharmaceutical preparation containing interferon |
| EP0080879B1 (en) * | 1981-11-28 | 1986-10-01 | Sunstar Kabushiki Kaisha | Pharmaceutical composition containing interferon in stable state |
| DE3262575D1 (en) * | 1981-12-23 | 1985-04-18 | Schering Corp | Stabilised interferon formulations and their preparation |
| ZA828580B (en) * | 1981-12-23 | 1983-10-26 | Schering Corp | Interferon formulations |
| US4957734A (en) * | 1982-06-14 | 1990-09-18 | Exovir, Inc. | Treatment of certain skin malignancies and pre-malignant skin lesions, herpes zoster and psoriasis |
| US4486407A (en) * | 1983-02-28 | 1984-12-04 | Fumiaki Taguchi | Method for enhancing production of interferon |
| US5385738A (en) * | 1983-10-14 | 1995-01-31 | Sumitomo Pharmaceuticals Company, Ltd. | Sustained-release injection |
| US4855134A (en) * | 1983-10-14 | 1989-08-08 | Sumitomo Pharmaceuticals Company, Limited | Sustained-release preparation |
| US4680175A (en) * | 1984-02-07 | 1987-07-14 | Interferon Sciences, Inc. | Interferon administration vehicles |
| WO1986000531A1 (en) * | 1984-07-10 | 1986-01-30 | Takeda Chemical Industries, Ltd. | Gamma-interferon composition |
| JPS61277633A (en) * | 1985-05-31 | 1986-12-08 | Toray Ind Inc | Interferon composition |
| JP2577742B2 (en) * | 1986-07-18 | 1997-02-05 | 中外製薬株式会社 | Stable granulocyte colony-stimulating factor containing preparation |
| JP2577744B2 (en) * | 1986-07-18 | 1997-02-05 | 中外製薬株式会社 | Stable granulocyte colony-stimulating factor containing preparation |
| CA1294215C (en) * | 1986-10-27 | 1992-01-14 | Ze'ev Shaked | Pharmaceutical compositions of recombinant beta-interferon and formulation processes |
| US4888416A (en) * | 1987-03-30 | 1989-12-19 | International Minerals & Chemical Corp. | Method for stabilizing somatotropins |
| DE10149030A1 (en) * | 2001-10-05 | 2003-04-10 | Viscum Ag | Preparation of storage-stable medicaments containing recombinant carbohydrate-binding polypeptides, especially r-viscumin, useful e.g. as cytotoxic agent, comprises cooling, freezing, spray-drying or lyophilizing solution of pH more than 6 |
| JP4610154B2 (en) * | 2002-05-30 | 2011-01-12 | 大塚製薬株式会社 | Injectable preparation |
| DE102006030164A1 (en) * | 2006-06-29 | 2008-01-03 | Boehringer Ingelheim Pharma Gmbh & Co. Kg | Inhalative powders |
| AR080428A1 (en) | 2010-01-20 | 2012-04-11 | Chugai Pharmaceutical Co Ltd | FORMULATIONS STABILIZED LIQUID CONTAINERS OF ANTIBODIES |
| EP4159236A4 (en) | 2020-05-29 | 2024-08-21 | Chugai Seiyaku Kabushiki Kaisha | FORMULATION CONTAINING AN ANTIBODY |
-
1979
- 1979-01-31 JP JP993979A patent/JPS55102519A/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04200105A (en) * | 1990-11-29 | 1992-07-21 | Nec Corp | Travelling wave tube power amplifier |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS55102519A (en) | 1980-08-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JPS6326120B2 (en) | ||
| US4847079A (en) | Biologically stable interferon compositions comprising thimerosal | |
| DE69432179T2 (en) | A FORMULATION OF THE COagulation Factor VIII | |
| JP3530300B2 (en) | Stable transglutaminase formulations and methods for producing them | |
| JP3143470B2 (en) | Pharmaceutical composition containing polypeptide for subcutaneous or intramuscular administration | |
| NL7905220A (en) | ANTI-THROMBINE PREPARATION AND METHOD FOR PREPARING IT. | |
| CA2288143A1 (en) | Activated protein c formulations | |
| JPS61103836A (en) | Fibronectin pharmaceutical preparation | |
| JPH0378373B2 (en) | ||
| JPH09510736A (en) | Heparin-based antithrombotic non-hemorrhagic composition, method for its preparation and therapeutic use | |
| JPS592686A (en) | Powder pharmaceutical preparation of myeloperoxidase | |
| JP2007528842A (en) | Stable human erythropoietin aqueous solution without serum albumin | |
| JP2003514027A (en) | α-interferon aqueous formulation | |
| JPH0380772B2 (en) | ||
| JPS63145279A (en) | Drug product | |
| HU205555B (en) | Process for producing stabilized leukocyte interferons | |
| RU2077336C1 (en) | Genetic-engineering gamma-interferon preparation | |
| EP0702960A1 (en) | Liquid globulin composition for intravenous injection, process for producing the same, and method of inhibiting globulin dimer increase in said composition | |
| JP2001000179A (en) | How to inactivate viruses | |
| DE69233315T2 (en) | PHARMACEUTICAL LYSINE-CONTAINING POLYPEPTIDE COMPOSITIONS AND METHOD FOR THEIR USE | |
| JP3820607B2 (en) | Antithrombin-III liquid preparation and storage stabilization method thereof | |
| JP2770911B2 (en) | Antiviral pharmaceutical composition | |
| JPS62153224A (en) | Plasminogen preparation | |
| JP3494667B2 (en) | Antithrombin III preparation | |
| WO1998040096A1 (en) | Preventives and/or remedies for multiple organ failure |