JPH01186818A - Drug preparation of complex of polyriboinosinic acid, polyribocytidylic acid and poly-l-lysine - Google Patents
Drug preparation of complex of polyriboinosinic acid, polyribocytidylic acid and poly-l-lysineInfo
- Publication number
- JPH01186818A JPH01186818A JP813188A JP813188A JPH01186818A JP H01186818 A JPH01186818 A JP H01186818A JP 813188 A JP813188 A JP 813188A JP 813188 A JP813188 A JP 813188A JP H01186818 A JPH01186818 A JP H01186818A
- Authority
- JP
- Japan
- Prior art keywords
- poly
- acid
- lysine
- aqueous solution
- polysaccharide
- 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.)
- Pending
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 239000002253 acid Substances 0.000 title claims abstract description 16
- 229920000729 poly(L-lysine) polymer Polymers 0.000 title claims abstract description 15
- 229940079593 drug Drugs 0.000 title abstract 3
- 239000003814 drug Substances 0.000 title abstract 3
- 108010010803 Gelatin Proteins 0.000 claims abstract description 18
- 239000008273 gelatin Substances 0.000 claims abstract description 18
- 229920000159 gelatin Polymers 0.000 claims abstract description 18
- 235000019322 gelatine Nutrition 0.000 claims abstract description 18
- 235000011852 gelatine desserts Nutrition 0.000 claims abstract description 18
- 229920001282 polysaccharide Polymers 0.000 claims abstract description 17
- 239000005017 polysaccharide Substances 0.000 claims abstract description 17
- 239000000203 mixture Substances 0.000 claims abstract description 15
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 15
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 15
- 102000008186 Collagen Human genes 0.000 claims abstract description 5
- 108010035532 Collagen Proteins 0.000 claims abstract description 5
- 229920002307 Dextran Polymers 0.000 claims abstract description 5
- 229920001436 collagen Polymers 0.000 claims abstract description 5
- 108010071390 Serum Albumin Proteins 0.000 claims abstract description 4
- 102000007562 Serum Albumin Human genes 0.000 claims abstract description 4
- 239000000470 constituent Substances 0.000 claims abstract description 4
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 claims abstract description 3
- 229920001287 Chondroitin sulfate Polymers 0.000 claims abstract description 3
- 229920001353 Dextrin Polymers 0.000 claims abstract description 3
- 239000004375 Dextrin Substances 0.000 claims abstract description 3
- 229940059329 chondroitin sulfate Drugs 0.000 claims abstract description 3
- 235000019425 dextrin Nutrition 0.000 claims abstract description 3
- 150000004676 glycans Chemical class 0.000 claims abstract 7
- 239000002246 antineoplastic agent Substances 0.000 abstract description 2
- 239000007864 aqueous solution Substances 0.000 description 31
- 238000010828 elution Methods 0.000 description 10
- 238000009472 formulation Methods 0.000 description 10
- 150000004804 polysaccharides Chemical class 0.000 description 10
- 239000012153 distilled water Substances 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 235000018102 proteins Nutrition 0.000 description 8
- 238000004108 freeze drying Methods 0.000 description 6
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 5
- 229930195725 Mannitol Natural products 0.000 description 5
- 230000001939 inductive effect Effects 0.000 description 5
- 239000000594 mannitol Substances 0.000 description 5
- 235000010355 mannitol Nutrition 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 2
- 241000288906 Primates Species 0.000 description 2
- 102000006382 Ribonucleases Human genes 0.000 description 2
- 108010083644 Ribonucleases Proteins 0.000 description 2
- 241000283984 Rodentia Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229920001284 acidic polysaccharide Polymers 0.000 description 2
- 150000004805 acidic polysaccharides Chemical class 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 159000000000 sodium salts Chemical class 0.000 description 2
- QCQCHGYLTSGIGX-GHXANHINSA-N 4-[[(3ar,5ar,5br,7ar,9s,11ar,11br,13as)-5a,5b,8,8,11a-pentamethyl-3a-[(5-methylpyridine-3-carbonyl)amino]-2-oxo-1-propan-2-yl-4,5,6,7,7a,9,10,11,11b,12,13,13a-dodecahydro-3h-cyclopenta[a]chrysen-9-yl]oxy]-2,2-dimethyl-4-oxobutanoic acid Chemical compound N([C@@]12CC[C@@]3(C)[C@]4(C)CC[C@H]5C(C)(C)[C@@H](OC(=O)CC(C)(C)C(O)=O)CC[C@]5(C)[C@H]4CC[C@@H]3C1=C(C(C2)=O)C(C)C)C(=O)C1=CN=CC(C)=C1 QCQCHGYLTSGIGX-GHXANHINSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical group OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 108010045569 atelocollagen Proteins 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- KXKPYJOVDUMHGS-OSRGNVMNSA-N chondroitin sulfate Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](OS(O)(=O)=O)[C@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](C(O)=O)O1 KXKPYJOVDUMHGS-OSRGNVMNSA-N 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- -1 dextran and dextrin Chemical class 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229960001438 immunostimulant agent Drugs 0.000 description 1
- 239000003022 immunostimulating agent Substances 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 239000012931 lyophilized formulation Substances 0.000 description 1
- 235000018977 lysine Nutrition 0.000 description 1
- 210000003739 neck Anatomy 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 229920000728 polyester Polymers 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
Landscapes
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
【発明の詳細な説明】
本発明はポリリボイノシン酸・ポリリボシチジル酸・ポ
リ−Ｌ−リジン複合体（以下、ポリ（ＩＣＬ）と省略す
る。）製剤に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a polyriboinosinic acid/polyribocytidylic acid/poly-L-lysine complex (hereinafter abbreviated as poly(ICL)) preparation.
インターフェロン（以下、ＩＦＮと省略する。）誘発物
質として知られるポリリボイノシン酸・ポリリボシチジ
ル酸（以下、ポリ（Ｉ） ・ポリ（Ｃ）と省略する。Polyriboinosinic acid/polyribocytidylic acid (hereinafter abbreviated as poly(I)/poly(C)) is known as an interferon (hereinafter abbreviated as IFN) inducing substance.
）は、各種合成ポリヌクレオチドの中では最も高い誘発
活性を有する。しかし、ウサギ及びマウスなどのげっ歯
頚においては高いＩ ＦＮ誘発活性を示すものの、ヒト
をはじめとする霊長類におけるその誘発活性は極めて低
い。この原因は、霊長類ではポリ（Ｉ） ・ポリ（Ｃ
）を分解する酵素である血中リボヌクレアーゼの活性が
、げっ書類に比べて極めて高く、ポリ（Ｉ） ・ポリ
（Ｃ）が体内で容易に分解されてしまうためであると考
えられている。このため、ポリ（Ｉ） ・ポリ（Ｃ）
にリボヌクレアーゼ抵抗性を付与するための試みが活発
に展開され、多糖類やポリペプチドとの複合体が種々開
発されてきた。ポリ（ＩＣも高いＩＦＮ誘発活性を示す
ことがら、有用な抗腫瘍剤があるいは免疫賦活剤として
期待されている。) has the highest inducing activity among various synthetic polynucleotides. However, although it shows high IFN-inducing activity in the necks of rodents such as rabbits and mice, its inducing activity in primates including humans is extremely low. The cause of this is poly(I) and poly(C) in primates.
This is thought to be due to the fact that the activity of blood ribonuclease, an enzyme that decomposes poly(I) and poly(C), is extremely high compared to that of rodents, and poly(I) and poly(C) are easily degraded in the body. For this reason, poly(I) ・poly(C)
Efforts have been made actively to impart ribonuclease resistance to polysaccharides, and various complexes with polysaccharides and polypeptides have been developed. Since poly(IC) also exhibits high IFN-inducing activity, it is expected to be a useful antitumor agent or immunostimulant.
（従来技術〕
ポリ（ＩＣＬ）は、ポリ（Ｉ） ・ポリ（Ｃ）水溶液
とポリーＬ−リジン水溶液を混合することにより調製さ
れ、通常ポリ（ＩＣＬ）水溶液として保存される。しか
し、ポリ（ＴＣＬ）水溶液は不安定であり、例えば、高
速ゲルバーミエイシミンクロマトグラフィー（以下、Ｉ
Ｉ Ｐ −Ｇ Ｐ Ｃと省略する。）において、経時的
な分子量分布の変化が観察されるなど、保存中、徐々に
変質する。(Prior Art) Poly(ICL) is prepared by mixing poly(I)/poly(C) aqueous solution and poly-L-lysine aqueous solution, and is usually stored as a poly(ICL) aqueous solution. However, poly(TCL) ) Aqueous solutions are unstable, for example, high-performance gel vermi-acimine chromatography (hereinafter referred to as I
It is abbreviated as IP-GPC. ) undergoes gradual deterioration during storage, with changes in molecular weight distribution observed over time.
ポリ（ＩＣＴ、）のＩＦＮ誘発活性は、ポリ（ＩＣＬ）
中のポリ（Ｉ）及びポリ（Ｃ）の分子量分布と、ポリ（
ＩＣＬ）の高次構造に依存すると考えられることから、
保存中の分子竜分布雫経時的変化は、ＩＦＮｌｖ発活性
に影響を及ぼす恐れがある。このため、保存安定性の優
れた製剤の開発が望まれている。The IFN-induced activity of poly(ICT,) is similar to that of poly(ICL).
Molecular weight distribution of poly(I) and poly(C) in poly(I) and poly(C)
Since it is thought to depend on the higher-order structure of ICL),
Changes in molecular distribution over time during storage may affect IFNlv activation activity. Therefore, it is desired to develop a formulation with excellent storage stability.
しばしば凍結乾燥の手段が用いられる。ポリ（Ｉ）・ポ
リ（Ｃ）及びポリーＬ−リジンについては、凍結乾燥品
として入手可能であり、これらの水溶液を凍結乾燥する
ことは従来技術に属する。しかしながら、ポリ（ＩＣＬ
）については、特開昭５６一−５３６２１号及び特開昭
８１−１０３８２４号明細書に記載されているように、
ポリ（ＩＣＬ）水溶液の調製過程において、不溶性の沈
澱が生じやすい、さらにポリ（ＩＣＬ）水溶液を凍結乾
燥することによる製剤化は、実用上非常に困薙なことと
考えられていた。実際、ポリ（ＩＣＬ）水溶液を凍結乾
燥した後、蒸留水を加えて再溶解しても澄明な水溶液に
はならないため、実用に供し得るものではなかった。Frequently, lyophilization procedures are used. Poly(I)-poly(C) and poly-L-lysine are available as freeze-dried products, and freeze-drying their aqueous solutions belongs to the prior art. However, poly(ICL)
), as described in JP-A-561-53621 and JP-A-81-103824,
In the process of preparing an aqueous poly(ICL) solution, insoluble precipitates are likely to occur, and furthermore, it has been thought that formulation by freeze-drying an aqueous poly(ICL) solution is extremely difficult in practice. In fact, even if a poly(ICL) aqueous solution is freeze-dried and then redissolved by adding distilled water, a clear aqueous solution cannot be obtained, so it could not be put to practical use.
本発明者らはポリ（ＩＣＬ）の凍結乾燥製剤の開発に取
り組み、マンニトール等の水溶性物質を添加して凍結乾
燥されたポリ（ＩＣＬ）は再溶解により澄明な水溶液に
再生することを見出した。The present inventors worked on developing a freeze-dried preparation of poly(ICL) and found that poly(ICL) that had been freeze-dried by adding a water-soluble substance such as mannitol could be regenerated into a clear aqueous solution by redissolving it. .
しかし、ポリ（ＩＣＬ）を凍結乾燥製剤とする際、通常
用いられる賦形剤、例えばマンニトール等を添加して凍
結乾燥製剤とした場合には長期間の保存により、再溶解
時の澄明性が低下するなど、溶解性が悪くなるという欠
点を有していた。However, when poly(ICL) is made into a lyophilized preparation by adding commonly used excipients such as mannitol, the clarity upon redissolution decreases due to long-term storage. It had the disadvantage of poor solubility.
本発明者らは、これらの欠点を改善する目的で鋭意検討
した結果、意外にも可溶性蛋白及び／又は多糖類等の特
定の物質を加えて凍結乾燥することにより、長期保存後
の溶解性及び安定性が著しく向上することを見出し１本
発明を完成するに至った・
すなわち、本発明は、ポリリボイノシン酸・ポリリボシ
チジル酸・ポリ−Ｌ−リジン複合体の凍結乾燥製剤であ
って、構成成分として可溶性蛋白及び／又は多糖類を含
有することを特徴とする凍結乾燥製剤に関する。As a result of intensive studies aimed at improving these drawbacks, the present inventors unexpectedly found that by adding specific substances such as soluble proteins and/or polysaccharides and freeze-drying, the solubility and solubility after long-term storage could be improved. They found that the stability was significantly improved and completed the present invention. Namely, the present invention is a lyophilized preparation of a polyriboinosinic acid/polyribocytidylic acid/poly-L-lysine complex, which comprises the constituent components: The present invention relates to a freeze-dried preparation characterized by containing a soluble protein and/or a polysaccharide.
本発明で使用するポリ（Ｉ） ・ポリ（Ｃ）は。The poly(I) and poly(C) used in the present invention are.
沈降定数がいずれも４８〜１１ｇであるポリ（Ｉ）とポ
リ（Ｃ）とから構成され、ヌクレオチドとしてのモル比
が１．０：０．５〜２．０、好ましくは１．０：０．８
〜１．２の複合体である。ポリ（Ｉ） ・ポリ（Ｃ）
は通常ナトリウム塩として用いられ、水溶液又は凍結乾
燥品のいずれの形態であっても良い。It is composed of poly(I) and poly(C), both of which have a sedimentation constant of 48 to 11 g, and the molar ratio of nucleotides is 1.0:0.5 to 2.0, preferably 1.0:0. 8
~1.2 complex. Poly(I) ・Poly(C)
is usually used as a sodium salt, and may be in the form of an aqueous solution or a lyophilized product.
ポリーＬ−リジンは１通常その臭化水素酸塩又は塩化水
素酸塩が用いられ、平均分子量はｓ、ｏｏ。Poly-L-lysine is usually used as its hydrobromide or hydrochloride salt, and has an average molecular weight of s, oo.
〜ｓｏ、ｏｏｏのものが好ましい。~so, ooo are preferred.
本発明に適用される可溶性蛋白及び多糖類は、ポリ（Ｉ
ＣＬ）凍結乾燥製剤及びその保存における安定性を向上
しうる性質を有するものである限り、特に限定されない
が、例えば可溶性蛋白としてはゼラチン、コラーゲン及
び血清アルブミン等が、又、多糖類としては中性多糖類
であるデキストラン及びデキストリン等、並びに酸性多
糖類であるコンドロイチン硫酸等が挙げられる。Soluble proteins and polysaccharides applicable to the present invention include poly(I)
CL) There are no particular limitations as long as they have properties that can improve the stability of freeze-dried preparations and their storage, but examples of soluble proteins include gelatin, collagen, and serum albumin, and examples of polysaccharides include neutral ones. Examples include polysaccharides such as dextran and dextrin, and acidic polysaccharides such as chondroitin sulfate.
ゼラチンは、酸処理ゼラチンが好ましいが特に限定され
ない、コラーゲンは、抗原性の点がχヒト由来のものが
好ましいが、入手が容易なアテロコラーゲン（コラーゲ
ンの末端部位に存在するペプチドを化学的に削除した〜
もの、）であっても良い、又、血清アルブミンは牛血清
アルブミン及びヒト血清アルブミンのいずれであっても
良いが。Gelatin is preferably acid-treated gelatin, but is not particularly limited. Collagen is preferably derived from humans in terms of antigenicity, but atelocollagen, which is easily available (in which peptides present in the terminal parts of collagen are chemically deleted), is preferred. ~
), and the serum albumin may be either bovine serum albumin or human serum albumin.
精製が充分でないとりボヌクレアーゼ活性を有すること
があるので充分精製されたものであることが望ましい、
又、フンドロイチン硫酸等の酸性多糖類は通常ナトリウ
ム塩及びカリウム塩が用いられるが、特に限定されない
、 。It is desirable that the product is sufficiently purified as it may have bonuclease activity if it is not purified sufficiently.
Further, acidic polysaccharides such as fundroitin sulfate are usually used as sodium salts and potassium salts, but are not particularly limited.
可溶性蛋白の添加量は、ポリ（ＩＣＬ）１重量部に対し
て０．３〜３０重量部が好ましく、又、多糖類の添加量
は、ポリ（ＩＣＬ）１重量部に対して０．１〜３０ｍ轍
部であることが好ましい。The amount of soluble protein added is preferably 0.3 to 30 parts by weight per 1 part by weight of poly(ICL), and the amount of polysaccharide added is preferably 0.1 to 30 parts by weight per 1 part by weight of poly(ICL). A 30m rut is preferred.
又、可溶性蛋白と多糖類を併用する場會の両者の使用比
率は、その種類に応じて任意に選択決定すれば良い０例
えば、可溶性蛋白：多糖類＝１＝０．１〜２０に調製し
たものをポリ（ＩＣＬ）１重量部に対して０．３〜３０
重量部添加するものが好ましい。In addition, when soluble protein and polysaccharide are used together, the ratio of both may be arbitrarily selected depending on the type. For example, soluble protein: polysaccharide = 1 = 0.1 to 20. 0.3 to 30 parts by weight of poly(ICL)
Preferably, it is added in parts by weight.
本発明の凍結乾燥製剤を製造するには、まず、ポリ（Ｉ
） ・ポリ（Ｃ）に必要ならば注射用蒸留水を加えて
０．１〜５．０■／−の水溶液とする。To produce the lyophilized preparation of the present invention, first, poly(I)
) If necessary, add distilled water for injection to poly(C) to make an aqueous solution of 0.1 to 5.0 μ/-.
別に、ポリーＬ−リジンの臭化水素酸塩を注射用蒸留水
に溶解し、０．２〜２．０ｍｇ／ａＱの水溶液とする０
次に、ポリ（Ｉ） ・ポリ（Ｃ）水溶液に、ポリーＬ
−リジン水溶液を徐々に添加し、撹拌することによりポ
リ（ＩＣＬ）水溶液を調製する。Separately, dissolve poly-L-lysine hydrobromide in distilled water for injection to make an aqueous solution of 0.2 to 2.0 mg/aQ.
Next, poly L was added to the poly(I)/poly(C) aqueous solution.
- Prepare a poly(ICL) aqueous solution by gradually adding the lysine aqueous solution and stirring.
この際、ポリ（Ｉ） ・ポリ（Ｃ）とポリーＬ−リジ
ンの混合比は、ポリ（Ｉ） ・ポリ（Ｃ）のリン酸基
とポリーＬ−リジンのＣ位のアミノ基のモル比が１．０
：０．５〜２．０であることが好ましい、得られたポリ
（ＩＣＬ）水溶液に、あらがじめ注射用蒸留水に溶解し
た可溶性蛋白及び／又は多等類の水溶液を混合し、必要
ならばメンブランフィルタ−で濾過した後、アンプル又
はバイアルビンに充填し、常法に従って凍結乾燥して密
封すれば良い。At this time, the mixing ratio of poly(I)-poly(C) and poly-L-lysine is such that the molar ratio of the phosphoric acid group of poly(I)-poly(C) and the amino group at the C position of poly-L-lysine is 1.0
: The obtained poly(ICL) aqueous solution, which is preferably 0.5 to 2.0, is mixed with an aqueous solution of a soluble protein and/or polyester previously dissolved in distilled water for injection. In that case, after filtration with a membrane filter, it may be filled into ampoules or vials, freeze-dried and sealed according to a conventional method.
なお本発明の製剤には所望により塩酸、水酸化ナトリウ
ム、塩化ナトリウム、リン酸ナトリウム、ブドウ糖、果
糖及びマンニトール等の通常の添加剤を加えることがで
きる。又、ｐＨは３〜９の範囲〔実施例〕
実施例 １
沈降定数６８のポリ（Ｉ）と９８のポリ（Ｃ）で、ヌク
レオチドとして等モルよりなるポリ（Ｉ）・ポリ（Ｃ）
の凍結乾燥品６７■を注射用蒸留水３０ａＧに溶解し、
ポリ（Ｉ） ・ポリ（Ｃ）水溶液とした。別に、平均
分子量３４，０００のポリーＬ−リジンの臭化水素酸塩
２７■を注射用蒸留水３〇−に溶解し、ポリーＬ−リジ
ン水溶液とした０次に。Note that conventional additives such as hydrochloric acid, sodium hydroxide, sodium chloride, sodium phosphate, glucose, fructose, and mannitol can be added to the preparation of the present invention, if desired. In addition, the pH is in the range of 3 to 9 [Example] Example 1 Poly(I) and poly(C) having a sedimentation constant of 68 and poly(C) having an equal mole of nucleotides.
Dissolve 67μ of the freeze-dried product in 30aG of distilled water for injection,
A poly(I)/poly(C) aqueous solution was prepared. Separately, 27 ml of hydrobromide of poly L-lysine having an average molecular weight of 34,000 was dissolved in 30 ml of distilled water for injection to prepare an aqueous solution of poly L-lysine.
ポリ（Ｉ） ・ポリ（Ｃ）水溶液にポリーＬ−リジン
水溶液を徐々に注入した。得られたポリ（ＩＣＬ）水溶
液に、平均分子ｍ ｓ 、 ｏｏｏの酸処理ゼラチンの
５％水溶液６−を混合し、孔径０．２２ミクロンのメン
ブランフィルタ−で無菌濾過した。その後バイアルビン
に充填し、凍結乾燥して密封し。Poly L-lysine aqueous solution was gradually injected into poly(I)/poly(C) aqueous solution. A 5% aqueous solution 6- of acid-treated gelatin having an average molecular weight of m s and ooo was mixed with the obtained poly(ICL) aqueous solution, and the mixture was sterile-filtered using a membrane filter with a pore size of 0.22 microns. It is then filled into vials, freeze-dried, and sealed.
凍結乾燥製剤とした。It was made into a lyophilized formulation.
実施例 ２
実施例１で用いた５％ゼラチン水溶液の代わりに、平均
分子量４０，０００のデキストランの１０％水製剤とし
た。Example 2 Instead of the 5% aqueous gelatin solution used in Example 1, a 10% aqueous preparation of dextran with an average molecular weight of 40,000 was used.
実施例 ３
実施例１で用いた５％ゼラチン水溶液の代わりに、２％
ゼラチンを含む２０％マンニトール水溶液を用いて、実
施例１と同様に操作し凍結乾燥製剤とした。Example 3 Instead of the 5% gelatin aqueous solution used in Example 1, 2%
A lyophilized preparation was prepared in the same manner as in Example 1 using a 20% mannitol aqueous solution containing gelatin.
実施例 ４
実施例１で用いた５％ゼラチン水溶液の代わりに、５％
牛血清アルブミン水溶液を用いて、実施例１と同様に操
作し凍結乾燥製剤とした。Example 4 Instead of the 5% gelatin aqueous solution used in Example 1, 5%
A lyophilized preparation was prepared in the same manner as in Example 1 using an aqueous bovine serum albumin solution.
実施例 ５
実施例１で用いた５％ゼラチン水溶液の代わりに、１０
％コンドロイチン硫酸ナトリウム水溶液を用いて、実施
例１と同様に操作し凍結乾燥製剤とした。Example 5 Instead of the 5% gelatin aqueous solution used in Example 1, 10
A freeze-dried preparation was prepared in the same manner as in Example 1 using a % sodium chondroitin sulfate aqueous solution.
実施例 ６
実施例１で用いた５％ゼラチン水溶液の代わりに、２％
のゼラチンを含む１０％デキストラン水溶液を用いて、
実施例１と同様に操作し凍結乾燥製剤とした。Example 6 Instead of the 5% aqueous gelatin solution used in Example 1, 2%
Using a 10% dextran aqueous solution containing gelatin,
The same procedure as in Example 1 was carried out to prepare a freeze-dried preparation.
実施例 ７
実施例１で用いた５％ゼラチン水溶液の代わりに、２％
のゼラチンを含む１０％フンドロイチン硫酸ナトリウム
水溶液を用いて、実施例１と同様に操作し凍結乾燥製剤
とした。Example 7 Instead of the 5% aqueous gelatin solution used in Example 1, 2%
A lyophilized preparation was prepared in the same manner as in Example 1 using a 10% sodium fundroitin sulfate aqueous solution containing gelatin.
比較例
実施例１で用いた５％ゼラチン水溶液の代わりに、２０
％マンニトール水溶液を用いて、実施例１と同様に操作
し凍結乾燥製剤とした。Comparative Example Instead of the 5% gelatin aqueous solution used in Example 1, 20
% mannitol aqueous solution and operated in the same manner as in Example 1 to obtain a lyophilized preparation.
試験例
実施例１．２及び３で調製した本発明の製剤それぞれＡ
、Ｂ及びＣ並びに比較例で調製した比較製剤りの安定性
の比較を行った。即ち、各々の製剤を２５℃で６０日間
保存し、経時的に、注射用蒸留水で再溶解したときの澄
明度を、６５０ｎ■及び４２０ｎｍにおける吸光度を測
定することにより調べた。又、再溶解後のポリ（ＩＣＬ
）の分子量分布をＩＭＦ−ＧＰＣ用カタカラムいて測定
した。Test Examples Preparations of the invention prepared in Examples 1.2 and 3, respectively A
, B and C, and the comparative formulations prepared in Comparative Examples were compared for stability. That is, each formulation was stored at 25° C. for 60 days, and its clarity when redissolved with distilled water for injection over time was examined by measuring the absorbance at 650 nm and 420 nm. In addition, poly(ICL) after redissolution
) was measured using a catacolumn for IMF-GPC.
その結果を第１表、第１図及び第２図に示す、再低下が
みられ、分子量分布も変化した。それに対し、本発明の
製剤Ａ、Ｂ及びＣでは澄明性及び分子量分布が殆ど変化
せず、保存安定性が著しく改善されていることが判る。The results are shown in Table 1, Figures 1 and 2. A re-decrease was observed, and the molecular weight distribution also changed. In contrast, in formulations A, B, and C of the present invention, the clarity and molecular weight distribution hardly change, and it can be seen that the storage stability is significantly improved.
第１表 ポリ（ＩＣＬ）凍結乾燥品の再溶解後の澄明性Table 1: Clarity of lyophilized poly(ICL) after remelting
第１図は、本発明の製剤Ａ、Ｂを蒸留水で溶解した後の
ＨＰ−ＧＰＣ用カタカラムる溶出曲線である。横軸は溶
出時間、縦軸は２５４ｎｍにおけるＵＶ吸収である。又
、それ−Ｆ、ｏ ＩＩ剤の溶出曲線は。
下から凍結乾燥直後、保存後１４日目及び２力月目のも
のである。溶出曲線上のピークは、溶出時の早い方から
、メインピーク、セカンドピークとし、メインピークは
ポリ（ＩＣＬ）に由来するピークであり、セカンドピー
クは製剤中に含まれるゼラチン等に由来するものである
。
第２図は２本発明の製剤Ｃおよび比較製剤りを蒸留水で
溶解した後のＨＰ−ＧＰＣ用カタカラムる溶出曲線であ
る。横軸は溶出時間、縦軸は２５４ｎ■におけるＵＶ吸
収である。又、それぞれの製瘤の溶出曲線は、下から凍
結乾燥直後、保存後１４日目及び２力月目のものである
。溶出曲線上のピークは、溶出時の早い方から、メイン
ピーク、セカンドピークとし、メインビークはポリ（Ｉ
ＣＬ）に由来するピークであり、セカンドピークは製剤
中に含まれるゼラチン等に由来するものである。FIG. 1 is an elution curve for HP-GPC after dissolving formulations A and B of the present invention in distilled water. The horizontal axis is elution time, and the vertical axis is UV absorption at 254 nm. Also, the elution curve of agent F,o II is as follows. From the bottom, the images are immediately after freeze-drying, 14 days after storage, and 2 months after storage. The peaks on the elution curve are the main peak and the second peak, starting from the earliest one at the time of elution.The main peak is a peak derived from poly(ICL), and the second peak is derived from gelatin, etc. contained in the formulation. be. FIG. 2 is an elution curve for HP-GPC after dissolving Formulation C of the present invention and Comparative Formulation in distilled water. The horizontal axis is the elution time, and the vertical axis is the UV absorption at 254n. In addition, the elution curves of each lump are, from the bottom, immediately after freeze-drying, 14 days after storage, and 2 months after storage. The peaks on the elution curve are the main peak and the second peak, starting from the earliest peak at the time of elution, and the main peak is poly(I).
CL), and the second peak is derived from gelatin, etc. contained in the formulation.
Claims (3)
−Ｌ−リジン複合体の凍結乾燥製剤であって、構成成分
として可溶性蛋白及び／又は多糖類を含有することを特
徴とする凍結乾燥製剤。(1) A lyophilized preparation of a polyriboinosinic acid/polyribocytidylic acid/poly-L-lysine complex, which is characterized by containing soluble protein and/or polysaccharide as a constituent component.
ブミンから選ばれた１種又は２種以上の混合物である特
許請求の範囲第（１）項に記載の凍結乾燥製剤。(2) The lyophilized preparation according to claim (1), wherein the soluble protein is one or a mixture of two or more selected from gelatin, collagen, and serum albumin.
ロイチン硫酸から選ばれた１種又は２種以上の混合物で
ある特許請求の範囲第（１）項に記載の凍結乾燥製剤。(3) The lyophilized preparation according to claim (1), wherein the polysaccharide is one or a mixture of two or more selected from dextran, dextrin, and chondroitin sulfate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP813188A JPH01186818A (en) | 1988-01-18 | 1988-01-18 | Drug preparation of complex of polyriboinosinic acid, polyribocytidylic acid and poly-l-lysine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP813188A JPH01186818A (en) | 1988-01-18 | 1988-01-18 | Drug preparation of complex of polyriboinosinic acid, polyribocytidylic acid and poly-l-lysine |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH01186818A true JPH01186818A (en) | 1989-07-26 |
Family
ID=11684734
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP813188A Pending JPH01186818A (en) | 1988-01-18 | 1988-01-18 | Drug preparation of complex of polyriboinosinic acid, polyribocytidylic acid and poly-l-lysine |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH01186818A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006131023A1 (en) * | 2005-06-08 | 2006-12-14 | Newbiomed Pika Pte Ltd | Polyinosinic acid-polycytidylic acid-based adjuvant |
| US8303966B2 (en) | 2006-01-13 | 2012-11-06 | Yisheng Biopharma (Singapore) Pte. Ltd. | Immunogenic substances comprising a polyinosinic acid—polycytidilic acid based adjuvant |
-
1988
- 1988-01-18 JP JP813188A patent/JPH01186818A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006131023A1 (en) * | 2005-06-08 | 2006-12-14 | Newbiomed Pika Pte Ltd | Polyinosinic acid-polycytidylic acid-based adjuvant |
| US7838017B2 (en) | 2005-06-08 | 2010-11-23 | Newbiomed Pika Pte Ltd | Polyinosinic acid-polycytidylic acid-based adjuvant |
| US8303965B2 (en) | 2005-06-08 | 2012-11-06 | Yisheng Biopharma (Singapore) Pte. Ltd. | Polyinosinic acid-polycytidylic acid-based adjuvant |
| US8303966B2 (en) | 2006-01-13 | 2012-11-06 | Yisheng Biopharma (Singapore) Pte. Ltd. | Immunogenic substances comprising a polyinosinic acid—polycytidilic acid based adjuvant |
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