JPS6233208B2

JPS6233208B2 -

Info

Publication number: JPS6233208B2
Application number: JP51068675A
Authority: JP
Inventors: Shuji Ookubo
Original assignee: Kyowa Hakko Kogyo Co Ltd
Current assignee: KH Neochem Co Ltd
Priority date: 1976-06-14
Filing date: 1976-06-14
Publication date: 1987-07-20
Also published as: JPS52151731A

Description

[Detailed description of the invention]

本発明はＮアセチルムラミル―Ｌ―アラニル―
Ｄ―グルタミル―γ―メソ―ジアミノピメリン酸
（Mur NAc―Ｌ―Ala―Ｄ―Glu―γ―meso
DAP，以下単にPGと略記する）またはウリジン
ジホスホ―Ｎアセチルムラミル―Ｌ―アラニル―
Ｄ―グルタミル―γ―メソ―ジアミノピメリン酸
（UDP―Mur NAc―Ｌ―Ala―Ｄ―Glu―γ―
meso DAP，以下単にUDP―PGと略記する）を
有効成分とする抗腫瘍性組成物に関する。従来、腫瘍の治療あるいは予防のための多くの
薬物が知られているが、なお新規な抗腫瘍性物質
の開発が望まれている。本発明者は種々研究の結果前記したごときPG
またはUDP―PGが優れた抗腫瘍活性を示すこと
を見出し本発明を完成するに到つた。本発明にか
かる物質の類似体がアジユバンド活性を有し、Ｔ
―リンパ、Ｂ―リンパ、補体系、発熱原性白血球
に対する作用を有し、また抗腫瘍作用を有するこ
とは知られているが、本発明のPGおよびUDP―
PGが抗腫瘍性を示すことは本発明者らがはじめ
て見出した知見である。以下、本発明を更に詳細に説明する。本発明の組成物はPGまたはUDP―PGを有効成
分として含む組成物である。PGおよびUDP―PG
はバイオケミカル・アンド・バイオフイジカル・
リサーチ・コミユニケーシヨンズ（BBRC）
vol，59，No.４，p1317〜1325，1974に報告された
公知物質である。本発明に用いるPGおよびUDP―PGは公知の手
法に従つて種々の微生物の菌体から抽出して得ら
れる。本発明に用いるPGおよびUDP―PGは広範な微
生物群、たとえばエツシエリヒア、ビブリオ、サ
ルモネラ、シユードモナス、などのグラム陰性
菌、バチルス、クロストリデイウムなどのグラム
陽性菌、ミクソバクテリアレス、コリネバクテリ
ア、ノカルデイア、ミコバクテリウム、ミクロモ
ノスポラ、ミクロバイスポラ、ミクロポリスポラ
などに見出される。本発明実施例に具体的に用いたPGおよびUDP
―PGはバチルス・メガテリウムKY3361
（ATCC13402）の菌体から公知の手法（上記文献
に記載の手法）に従つて抽出されたもので、弱酸
性で、白色のものである。本発明にかかわるPGおよびUDP―PGは種々の
腫瘍に対する予防および治療の効果を有するが、
これら物質の薬理効果に関する実験例を以下に示
す。実験例１試験動物として約20ｇのCDFマウス１群４匹
を使用した。メチルコラントレン誘発肉腫細胞５
×10⁵個をマウスの腹腔内に移植し、移植する７
日前および移植から１日後に、PGの生理食塩水
溶解液（８mg／ml）0.5mlを腹腔内に投与した。
１回の投与量は200mg／Kgに相当する。PG投与群
（試験群）とPG無投与群（対照群）の生存日数は
第１表の通りであつた。 The present invention relates to N-acetylmuramyl-L-alanyl-
D-glutamyl-γ-meso-diaminopimelic acid (Mur NAc-L-Ala-D-Glu-γ-meso
DAP, hereinafter simply abbreviated as PG) or uridine diphospho-N-acetylmuramyl-L-alanyl-
D-glutamyl-γ-meso-diaminopimelic acid (UDP-Mur NAc-L-Ala-D-Glu-γ-
The present invention relates to an antitumor composition containing meso DAP (hereinafter simply abbreviated as UDP-PG) as an active ingredient. Although many drugs for treating or preventing tumors have been known, there is still a desire to develop new antitumor substances. As a result of various studies, the inventor has discovered the above-mentioned PG.
Furthermore, the present inventors discovered that UDP-PG exhibits excellent antitumor activity and completed the present invention. Analogues of the substances according to the present invention have adjuvant activity and T
-It is known that the PG and UDP of the present invention have effects on lymph, B-lymph, the complement system, and pyrogenic leukocytes, and also have antitumor effects.
The present inventors discovered for the first time that PG exhibits antitumor properties. The present invention will be explained in more detail below. The composition of the present invention is a composition containing PG or UDP-PG as an active ingredient. PG and UDP—PG
is biochemical and biophysical.
Research Communications (BBRC)
It is a known substance reported in Vol. 59, No. 4, p1317-1325, 1974. PG and UDP-PG used in the present invention are obtained by extraction from the cells of various microorganisms according to known techniques. PG and UDP-PG used in the present invention can be used in a wide range of microbial groups, including Gram-negative bacteria such as E. , Mycobacterium, Micromonospora, Microbispora, Micropolyspora, etc. PG and UDP specifically used in the examples of the present invention
-PG is Bacillus megaterium KY3361
(ATCC13402) according to a known method (method described in the above-mentioned literature), and is weakly acidic and white in color. Although the PG and UDP-PG related to the present invention have preventive and therapeutic effects on various tumors,
Experimental examples regarding the pharmacological effects of these substances are shown below. Experimental Example 1 A group of 4 CDF mice weighing approximately 20 g were used as test animals. Methylcholanthrene-induced sarcoma cells 5
Transplant ⁵ x 10 cells into the abdominal cavity of the mouse.7
One day before and one day after transplantation, 0.5 ml of a physiological saline solution (8 mg/ml) of PG was administered intraperitoneally.
One dose corresponds to 200mg/Kg. The survival days of the PG administration group (test group) and the PG non-administration group (control group) were as shown in Table 1.

【表】実施例２移植肉腫細胞数を2.5×10⁵個用いる以外は実験
例１と同様に行つて第２表に示す結果を得た。[Table] Example 2 The same procedure as in Experimental Example 1 was performed except that the number of transplanted sarcoma cells was 2.5×10 ⁵ cells, and the results shown in Table 2 were obtained.

【表】実験例３試験動物として約19ｇのddマウス１群６匹を
使用した。マウスにザルコーマ180肉腫細胞５×
10⁶個を腋窩部皮下に移植し、移植24時間後、３
日後および６日後はPGの生理食塩水溶液（８
mg／mlおよび２mg／ml）0.5mlを尾静脈内に投与
した。投与量は各々200mg／Kg／day、50mg／
Kg／dayに相当する。移植７日後の腫瘍の体積を
第３表に示す。対照群としてPG無投与群を設けた。[Table] Experimental Example 3 One group of 6 dd mice weighing approximately 19 g were used as test animals. Sarcoma 180 sarcoma cells 5x in mice
10 ⁶ pieces were transplanted subcutaneously in the axillary area, and 24 hours after transplantation, 3
After 1 day and 6 days, PG in physiological saline solution (8
mg/ml and 2 mg/ml) was administered into the tail vein. Dosage is 200mg/Kg/day and 50mg/day, respectively.
Equivalent to kg/day. Table 3 shows the tumor volume 7 days after transplantation. A group without PG administration was established as a control group.

【表】実験例４試験動物として約20ｇのCDFマウス１群５匹
を使用した。UDP―PGの生理食塩水溶解液（８
および２mg／ml）0.5mlをマウス腹腔内に投与す
る。投与量は各々200mg／Kg、50mg／Kgに相当す
る。投与７日後にメチルコラントレン誘発肉腫細
胞４×10⁵個をマウス腹腔内に移植した。移植後
の生存日数を第４表に示す。対照群としてUDP
―PG無投与群を設けた。[Table] Experimental Example 4 One group of 5 CDF mice weighing approximately 20 g was used as test animals. Physiological saline solution of UDP-PG (8
and 2 mg/ml) is administered intraperitoneally to mice. The doses correspond to 200 mg/Kg and 50 mg/Kg, respectively. Seven days after administration, 4 x 10 ⁵ methylcholanthrene-induced sarcoma cells were intraperitoneally transplanted into the mice. The survival days after transplantation are shown in Table 4. UDP as control group
-A PG non-administration group was established.

【表】実験例５試験動物として約20ｇのCDFマウス１群５匹
を使用した。UDP―PGの生理食塩水溶解液（32
および８mg／ml）0.5mlを腹腔内に投与する。投
与量は各々800mg／Kgおよび200mg／Kgに相当す
る。投与７日後にメチルコラントレン誘発肉腫細
胞５×10⁵個をマウス腹腔内に移植した。移植後
の生存日数を第５表に示す。対照群としてUDP
―PG無投与群を設けた。[Table] Experimental Example 5 One group of 5 CDF mice weighing approximately 20 g was used as test animals. Physiological saline solution of UDP-PG (32
and 8 mg/ml) 0.5 ml is administered intraperitoneally. The doses correspond to 800mg/Kg and 200mg/Kg respectively. Seven days after administration, 5 x 10 ⁵ methylcholanthrene-induced sarcoma cells were intraperitoneally transplanted into the mice. The survival days after transplantation are shown in Table 5. UDP as control group
-A PG non-administration group was established.

【表】実験例６ PGに替えてUDP―PGを用いるほかは実験例１
と同様行つて第６表に示す結果を得た。[Table] Experimental example 6 Experimental example 1 except that UDP-PG is used instead of PG
The same procedure as above was carried out, and the results shown in Table 6 were obtained.

【表】実験例７試験動物として約19ｇのddマウス１群６匹を
使用した。試験群(1)にはザルコーマ180肉腫細胞５×10⁶個
を腋窩部皮下に移植し、移植24時間後および３日
後にUDP―PGの８mg／ml生理食塩水溶解液0.5ml
を尾静脈内に投与した。また別の試験群(2)にはザ
ルコーマ肉腫細胞５×10⁶個を腋窩部皮下に移植
し、移植24時間後、３日後および６日後にUDP
―PGの２mg／ml生理食塩水溶解液0.5mlを尾静脈
内に投与した。移植７日後の腫瘍の体積を第７表
に示す。対照群としてUDP―PG無投与群を設け
た。[Table] Experimental Example 7 One group of 6 dd mice weighing approximately 19 g were used as test animals. In the test group (1), 5 x 10 ⁶ Sarcoma 180 sarcoma cells were subcutaneously transplanted into the axillary region, and 0.5 ml of an 8 mg/ml solution of UDP-PG in physiological saline was administered 24 hours and 3 days after transplantation.
was administered into the tail vein. In another test group (2), 5 × 10 ⁶ Sarcoma sarcoma cells were subcutaneously transplanted into the axillary region, and UDP was applied 24 hours, 3 days, and 6 days after transplantation.
-0.5 ml of a 2 mg/ml solution of PG in physiological saline was administered into the tail vein. Table 7 shows the tumor volume 7 days after transplantation. A UDP-PG non-administration group was established as a control group.

【表】以上の実験例に示したごとく本発明にかかる
PGおよびUDP―PGは腫瘍の予防および治療に優
れた効果を示すことがわかる。本発明のPGおよびUDP―PGの急性毒性
（LD₅₀）を測定した結果を以下の実験例８に示
す。実験例８ ddYマウス（体重約20ｇ）１群３匹に、PGお
よびUDP―PGの生理食塩水溶解液（128，64，32
mg／ml）0.5mlを尾静脈内に投与した。投与量は
各々3200，1600，800mg／Kgに相当する。投与後
14日間マウスの生死を観察した結果、PGおよび
UDP―PGの両者とも3200mg／Kg投与群では全例
死亡したが、1600mg／Kg、800mg／Kg投与群では
全例生存していた。この結果から本発明のPGお
よびUDP―PGのLD₅₀は約2400mg／Kgと推定され
る。人の腫瘍治療のために本発明のPGおよびUDP
―PGを用いるときは、PGまたはUDP―PGの投
与を主に静脈内投与によつて行う。静脈への投与
に際しては生理食塩水、リンゲル液、５％キシリ
ツトなどに溶かして使用する。投与量としては
5.8〜23mg／Kg／dayが適当である。以下に本発明組成物の具体例を実施例として示
す。実施例１ PG6ｇを生理食塩水100mlに溶かして、PGの生
理食塩水溶液をつくり、この10mlをアンプルに封
入してPGの注射液とする。実施例２ PG6ｇをリンゲル液100mlに溶かして、PGのリ
ンゲル液をつくり、この10mlをアンプルに封入し
PGの注射液とする。実施例３ PG6ｇを５％キシリツト液100mlに溶かして、
PGのキシリツト液をつくり、この10mlをアンプ
ルに封入してPGの注射液とする。実施例４ PGに替えてUDP―PGを用いるほかは実施例１
と同様にしてUDP―PGの注射液とする。実施例５ PGに替えてUDP―PGを用いるほかは実施例２
と同様にしてUDP―PGの注射液とする。実施例６ PGに替えてUDP―PGを用いるほかは実施例３
と同様にしてUDP―PGの注射液とする。[Table] As shown in the above experimental examples, according to the present invention
It can be seen that PG and UDP-PG exhibit excellent effects on tumor prevention and treatment. The results of measuring the acute toxicity (LD ₅₀ ) of PG and UDP-PG of the present invention are shown in Experimental Example 8 below. Experimental Example 8 A group of three ddY mice (weighing approximately 20 g) was given a physiological saline solution of PG and UDP-PG (128, 64, 32).
mg/ml) 0.5 ml was administered into the tail vein. The doses correspond to 3200, 1600, and 800 mg/Kg, respectively. After administration
As a result of observing the survival of mice for 14 days, PG and
For both UDP-PG, all cases died in the 3200mg/Kg administration group, but all cases survived in the 1600mg/Kg and 800mg/Kg administration groups. From this result, the LD ₅₀ of the PG and UDP-PG of the present invention is estimated to be approximately 2400 mg/Kg. PG and UDP of the present invention for human tumor treatment
- When using PG, PG or UDP-PG is primarily administered intravenously. For intravenous administration, it is used after being dissolved in physiological saline, Ringer's solution, 5% xylate, etc. As for the dosage
5.8 to 23 mg/Kg/day is appropriate. Specific examples of the composition of the present invention are shown below as examples. Example 1 A physiological saline solution of PG is prepared by dissolving 6 g of PG in 100 ml of physiological saline, and this 10 ml is sealed in an ampoule to prepare a PG injection solution. Example 2 PG Ringer's solution was prepared by dissolving 6 g of PG in 100 ml of Ringer's solution, and this 10 ml was sealed in an ampoule.
Use as PG injection solution. Example 3 Dissolve 6g of PG in 100ml of 5% xyrite solution,
Prepare a PG xyrite solution and seal 10 ml of this in an ampoule to make a PG injection solution. Example 4 Example 1 except that UDP-PG is used instead of PG
In the same manner as above, prepare a UDP-PG injection solution. Example 5 Example 2 except that UDP-PG is used instead of PG
In the same manner as above, prepare a UDP-PG injection solution. Example 6 Example 3 except that UDP-PG is used instead of PG
In the same manner as above, prepare a UDP-PG injection solution.

Claims

[Claims]

1 Antitumor properties containing N-acetylmuramyl-L-alanyl-D-glutamyl-γ-meso-diaminopimelic acid or uridine diphospho-N-acetylmuramyl-L-alanyl-D-glutamyl-γ-meso-diaminopimelic acid as an active ingredient Composition.