JPH0214331B2 - - Google Patents
Info
- Publication number
- JPH0214331B2 JPH0214331B2 JP57150733A JP15073382A JPH0214331B2 JP H0214331 B2 JPH0214331 B2 JP H0214331B2 JP 57150733 A JP57150733 A JP 57150733A JP 15073382 A JP15073382 A JP 15073382A JP H0214331 B2 JPH0214331 B2 JP H0214331B2
- Authority
- JP
- Japan
- Prior art keywords
- nisin
- mice
- group
- administered
- present
- 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
- NVNLLIYOARQCIX-MSHCCFNRSA-N Nisin Chemical compound N1C(=O)[C@@H](CC(C)C)NC(=O)C(=C)NC(=O)[C@@H]([C@H](C)CC)NC(=O)[C@@H](NC(=O)C(=C/C)/NC(=O)[C@H](N)[C@H](C)CC)CSC[C@@H]1C(=O)N[C@@H]1C(=O)N2CCC[C@@H]2C(=O)NCC(=O)N[C@@H](C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(NCC(=O)N[C@H](C)C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCSC)C(=O)NCC(=O)N[C@H](CS[C@@H]2C)C(=O)N[C@H](CC(N)=O)C(=O)N[C@H](CCSC)C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(N[C@H](C)C(=O)N[C@@H]3C(=O)N[C@@H](C(N[C@H](CC=4NC=NC=4)C(=O)N[C@H](CS[C@@H]3C)C(=O)N[C@H](CO)C(=O)N[C@H]([C@H](C)CC)C(=O)N[C@H](CC=3NC=NC=3)C(=O)N[C@H](C(C)C)C(=O)NC(=C)C(=O)N[C@H](CCCCN)C(O)=O)=O)CS[C@@H]2C)=O)=O)CS[C@@H]1C NVNLLIYOARQCIX-MSHCCFNRSA-N 0.000 claims description 28
- 108010053775 Nisin Proteins 0.000 claims description 28
- 239000004309 nisin Substances 0.000 claims description 28
- 235000010297 nisin Nutrition 0.000 claims description 28
- 239000002246 antineoplastic agent Substances 0.000 claims description 6
- 239000004480 active ingredient Substances 0.000 claims description 3
- 229940041181 antineoplastic drug Drugs 0.000 claims 1
- 241000699670 Mus sp. Species 0.000 description 11
- 206010028980 Neoplasm Diseases 0.000 description 9
- 201000011510 cancer Diseases 0.000 description 7
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- 206010003445 Ascites Diseases 0.000 description 5
- 239000002244 precipitate Substances 0.000 description 5
- 230000001093 anti-cancer Effects 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 241000194035 Lactococcus lactis Species 0.000 description 3
- 235000014897 Streptococcus lactis Nutrition 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- PAWSVPVNIXFKOS-UHFFFAOYSA-N 2-aminobut-2-enoic acid zwitterion Chemical compound CC=C([NH3+])C([O-])=O PAWSVPVNIXFKOS-UHFFFAOYSA-N 0.000 description 1
- UQBOJOOOTLPNST-UHFFFAOYSA-N Dehydroalanine Chemical compound NC(=C)C(O)=O UQBOJOOOTLPNST-UHFFFAOYSA-N 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000003560 cancer drug Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000019249 food preservative Nutrition 0.000 description 1
- 239000005452 food preservative Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical class CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 1
- 231100001231 less toxic Toxicity 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- SJWFXCIHNDVPSH-UHFFFAOYSA-N octan-2-ol Chemical compound CCCCCCC(C)O SJWFXCIHNDVPSH-UHFFFAOYSA-N 0.000 description 1
- 210000003200 peritoneal cavity Anatomy 0.000 description 1
- 239000003910 polypeptide antibiotic agent Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Description
【発明の詳細な説明】
本発明は新規な制癌剤に関するものである。
更に詳細には、本発明は、ストレプトコツカ
ス・ラクチス（Streptococcus lactis）の特定の
菌株によつて生産される抗菌性ペプチド物質であ
るナイシン（Nisin）を有効成分とする新規な制
癌剤に関するものである。
一般に、臨床上の癌は、その組織型、原発部
位、薬剤感受性、悪性度などで生物学的多様性を
有しており、それ故に一種類の薬剤でこれを治療
することは極めて困難であると言わねばならな
い。したがつて、新規な制癌剤の開発は医療上の
急務であり、産業上においてもまた甚大なる意義
を有するものである。
本発明者らは、新規制癌剤の開発を目的として
乳酸菌の代謝産物の制癌活性を鋭意検討した結
果、ストレプトコツカス・ラクチスの特定の菌株
によつて生産されるポリペプチドであるナイシン
に顕著なる制癌活性を見出し、本発明に至つたも
のである。
ナイシンは、下記に示すように34個のアミノ酸
から構成されるポリペプチドで（J.Amer.Chem.
Soc.93，4634〜4635，1971）、グラム陽性細菌に
対して抗菌力を示すことが報告されてはいるが、
抗菌スペクトルがせまく生理PHでの溶解性がよく
ないために抗菌剤としては臨床上の使用に至つて
いない。
ABA：アミノ酪酸（aminobutylic acid）
DHA：デヒドロアラニン
DHB：デヒドロブチリン（β―メチルデヒドロ
アラニン）
しかしながら、ナイシンは毒性が低くしかもク
ロストリデイウム・ブチリカム（Clostridium
butyricum）やクロストリデイウムチロブチリカ
ム（Cl.tyrobutyricum）のような有害菌の生育を
効果的に抑えるため、食品の保存剤としては有効
であり、ヨーロツパ諸国の中には食品添加物とし
ての使用を法的に認めている国もある。実際に、
東欧諸国では野菜類のかん詰にこのナイシンを利
用することが行われており、ポーランドやソ連邦
ではナイシンの工業生産が行われている
（Antibiotiki18，162〜165，1973）。
本発明はナイシンを有効成分とする制癌剤に関
するが、本発明に使用するナイシンとしては上述
のごとき市販品を使用することも可能であるが、
ストレプトコツカス・ラクチス ATCC11454株
のごときナイシン生産性の菌株からナイシンを単
離、精製して使用することも可能である。
ナイシンの単離、精製法の一例を示せば次のと
おりである。すなわち、ナイシン生産菌の倍養液
をPH２とした後、遠心分離して菌体を除去し、上
精をPH4.2〜4.5として５％のクロロホルムと0.1％
の２級オクチルアルコールで乳濁させる。この混
合物をしばらく静置すると比較的少量の安定なク
ロロホルムゲルを生じ、活性はこのゲルに存在す
る。このゲルにアルコールを加えてクロロホルム
を溶解すると、活性物質の沈殿が得られる。この
沈殿を希酸中に溶解してPH2.5〜３に調整し、等
量のアルコールを加えることによつて不純物を沈
殿として除去した後、アルコール溶液を減圧下で
濃縮し、PH４に調整して冷却すると活性物質が沈
殿してくる。この沈殿を1/100規定の塩酸に溶解
して凍結乾燥すると、１ｇ当たり106〜２×106リ
ーデイングユニツト（Reading Unit）の力価を
有するナイシンが得られる。なおリーデイングユ
ニツトとは、ナイシンの標準力価のことで、トレ
イマー（Tramer）らによつて定められ、WHO
にも承認されている（1969年）。
本発明にかかるナイシンの制癌活性は、エール
リツヒ腹水腫瘍を用いて判定された。すなわち、
１群10匹のddYマウスにエールリツヒ腹水腫瘍細
胞を腹腔内接種し、翌日から連続９日間毎日ナイ
シンの投与を続け、対照群には生理食塩水の投与
を続けた。実験の詳細は実施例中に述べるが、ナ
イシン投与群と対照群との間には、体重の経時的
変化（第１図）および延命（実施例の表―１）に
顕著な差異が認められた。なお第１図において、
ナイシン投与群Ａに比して対照群Ｂの体重の著し
い増加は、エールリツヒ腫瘍細胞の増殖に伴う腹
水の貯留を反映したものである。
本実験では、通常のddYマウスを発癌させるの
に十分なエールリツヒ腫瘍細胞数を接種したにも
拘らず、ナイシン投与時では癌細胞の腹腔内での
増殖がまつたく認められないか、発癌しないマウ
スが大半を占めた。また発癌したマウスでも、ナ
イシン投与群のマウスは対照群に比べて顕著に延
命した。
以上の結果より、本発明者らは抗菌性ポリペプ
チド物質ナイシンに従来思いもよらなかつた新規
な制癌活性を確認し、ナイシンのかかる性質を利
用することを特徴とする有用な制癌剤を発明する
に到つた。本発明に用いるナイシンは毒性が著し
く低いため（Journal of the Science of Food
and Agriculture 13，32〜42，1962）、大量生体
投与が可能であり、長期間投与が可能である。投
与量は、１〜104mg／Kg／day程度であり、その
剤状は散剤、カプセル剤、水剤、乳剤などで、注
射薬、内服薬、坐薬などによつて投与される。
次に本発明の実施例を示す。
実施例
ddY系６週令の雌性マウスを用い、１群10匹と
し予め１週間ddYマウスに継代増殖したエールリ
ツヒ腹水腫瘍細胞を３×105個腹腔内に接種し、
翌日より連続９日間、対照群には生理食塩水を、
試験群には生理食塩水に溶解したナイシンをそれ
ぞれ腹腔内に投与した。ナイシンの投与量は40
mg／Kg／dayとしたが、本実験に用いたナイシン
は106ユニツト／ｇの力価のものであり、マウス
１匹１日当たりに投与されたナイシンの力価は約
1000ユニツトである。
対照群および試験群中で、第１匹目のマウスが
死亡するまで経時的に体重の測定を続けた。対照
群のマウスは10匹中のすべてが発癌し、腫瘍細胞
接種後35日目までにすべて死亡したのに対し、ナ
イシンを投与した試験群では、発癌は10匹中でわ
ずか２匹に認められたのみであつた。
結果は表―１に示される。
【表】DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel anticancer agent. More specifically, the present invention relates to a novel anticancer agent containing nisin, an antibacterial peptide substance produced by a specific strain of Streptococcus lactis, as an active ingredient. . In general, clinical cancers have biological diversity in terms of tissue type, primary site, drug sensitivity, malignancy, etc., and it is therefore extremely difficult to treat them with a single type of drug. I have to say that. Therefore, the development of new anticancer agents is an urgent medical necessity, and is also of great industrial significance. As a result of intensive investigation into the anticancer activity of metabolites of lactic acid bacteria for the purpose of developing new regulated cancer drugs, the present inventors found that nisin, a polypeptide produced by a specific strain of Streptococcus lactis It was discovered that it has anticancer activity, leading to the present invention. Nisin is a polypeptide composed of 34 amino acids as shown below (J.Amer.Chem.
Soc. 93, 4634-4635, 1971), although it has been reported that it exhibits antibacterial activity against Gram-positive bacteria.
It has not been used clinically as an antibacterial agent because its antibacterial spectrum is narrow and its solubility at physiological pH is poor. ABA: aminobutylic acid DHA: dehydroalanine DHB: dehydrobutyrin (β-methyldehydroalanine) However, nisin is less toxic and
It is effective as a food preservative because it effectively suppresses the growth of harmful bacteria such as Cl. Some countries legally allow its use. actually,
Nisin is used in canned vegetables in Eastern European countries, and industrial production of nisin is carried out in Poland and the Soviet Union (Antibiotiki 18 , 162-165, 1973). The present invention relates to an anticancer agent containing nisin as an active ingredient, but as the nisin used in the present invention, commercially available products such as those mentioned above can be used, but
It is also possible to isolate and purify nisin from a nisin-producing strain such as Streptococcus lactis ATCC11454 strain for use. An example of a method for isolating and purifying nisin is as follows. That is, after adjusting the culture solution of nisin-producing bacteria to pH 2, centrifuging to remove the bacterial cells, and adjusting the supernatant to pH 4.2 to 4.5, 5% chloroform and 0.1%
Emulsify with secondary octyl alcohol. When this mixture is allowed to stand for a while, it forms a relatively small amount of stable chloroform gel, in which the activity resides. When alcohol is added to this gel to dissolve the chloroform, a precipitate of the active substance is obtained. This precipitate was dissolved in dilute acid to adjust the pH to 2.5 to 3, and an equal amount of alcohol was added to remove impurities as a precipitate, and the alcohol solution was concentrated under reduced pressure and adjusted to pH 4. When cooled, the active substance will precipitate. When this precipitate is dissolved in 1/100 normal hydrochloric acid and lyophilized, nisin having a titer of 10 6 to 2×10 6 Reading Units per gram is obtained. The leading unit is the standard titer of nisin, determined by Tramer et al.
(1969). The anticancer activity of nisin according to the present invention was determined using Ehrrich ascites tumor. That is,
Ehrlitsu ascites tumor cells were intraperitoneally inoculated into 10 ddY mice per group, and administration of nisin was continued every day for 9 consecutive days starting from the next day, and administration of physiological saline was continued to the control group. The details of the experiment will be described in the Examples, but significant differences were observed between the nisin-administered group and the control group in changes in body weight over time (Figure 1) and survival prolongation (Table 1 in Examples). Ta. In addition, in Figure 1,
The significant increase in body weight in control group B compared to nisin-administered group A reflects the accumulation of ascites accompanying the proliferation of Ehrlichi tumor cells. In this experiment, despite inoculation with a sufficient number of Ehrlichi tumor cells to cause cancer in normal ddY mice, when nisin was administered, cancer cells did not rapidly proliferate in the peritoneal cavity, or mice did not develop cancer. accounted for the majority. Even among mice that developed cancer, the survival of mice in the nisin-treated group was significantly longer than in the control group. Based on the above results, the present inventors have confirmed that the antibacterial polypeptide substance nisin has a novel anticancer activity that was previously unsuspected, and has invented a useful anticancer agent that utilizes this property of nisin. I reached it. Nisin used in the present invention has extremely low toxicity (Journal of the Science of Food
and Agriculture 13, 32-42, 1962), large amounts can be administered to living organisms, and long-term administration is possible. The dosage is about 1 to 10 4 mg/Kg/day, and the dosage form is powder, capsule, solution, emulsion, etc., and it is administered by injection, oral medicine, suppository, etc. Next, examples of the present invention will be shown. Example Using 6-week-old female ddY mice, 10 mice per group, 3 x 10 5 Ehrlichi ascites tumor cells that had been subcultured into ddY mice for 1 week were inoculated intraperitoneally.
Starting from the next day, the control group received physiological saline for 9 consecutive days.
Nisin dissolved in physiological saline was intraperitoneally administered to each of the test groups. Nisin dosage is 40
mg/Kg/day, but the nisin used in this experiment had a titer of 10 6 units/g, and the titer of nisin administered per mouse per day was approximately
It is 1000 units. Body weight measurements continued over time in the control and test groups until the first mouse died. All 10 mice in the control group developed cancer and all died by day 35 after tumor cell inoculation, whereas in the test group that received nisin, only 2 out of 10 mice developed cancer. It was just a matter of time. The results are shown in Table-1. 【table】
第１図はddYマウスにエールリツヒ腹水腫瘍細
胞を腹腔内接種した後の体重変化を示す図であ
る。
Ａ…ナイシン投与群、Ｂ…対照群。
FIG. 1 shows changes in body weight after intraperitoneal inoculation of Ehrlichi ascites tumor cells into ddY mice. A: Nisin administration group, B: control group.
Claims (1)
特徴とする制癌剤。1. An anticancer drug characterized by containing nisin as an active ingredient.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP57150733A JPS5942322A (en) | 1982-09-01 | 1982-09-01 | Carcinostatic agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP57150733A JPS5942322A (en) | 1982-09-01 | 1982-09-01 | Carcinostatic agent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5942322A JPS5942322A (en) | 1984-03-08 |
| JPH0214331B2 true JPH0214331B2 (en) | 1990-04-06 |
Family
ID=15503220
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP57150733A Granted JPS5942322A (en) | 1982-09-01 | 1982-09-01 | Carcinostatic agent |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5942322A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0442630U (en) * | 1990-08-10 | 1992-04-10 |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ATE491725T1 (en) * | 2002-07-15 | 2011-01-15 | Univ Texas | ANTIBODIES BOUND TO ANIONIC PHOSPHOLIPIDES AND AMINOPHOSPHOLIPIDES AND THEIR USE IN THE TREATMENT OF VIRUS INFECTIONS |
| US7378386B2 (en) * | 2002-07-15 | 2008-05-27 | Board Of Regents, The University Of Texas System | Anti-viral treatment methods using phosphatidylethanolamine-binding peptide derivatives |
| CA2485178A1 (en) * | 2004-11-22 | 2006-05-22 | Universite Du Quebec A Chicoutimi | Lactic acid bacteria-derived bacteriocin and uses thereof for prevention or treatment of cancer |
-
1982
- 1982-09-01 JP JP57150733A patent/JPS5942322A/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0442630U (en) * | 1990-08-10 | 1992-04-10 |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5942322A (en) | 1984-03-08 |
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