JPS6233209B2

JPS6233209B2 -

Info

Publication number: JPS6233209B2
Application number: JP51068676A
Authority: JP
Inventors: Masanori Fukui
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: JPS52151732A

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
ジホスホー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UDP―PGと略記する）を有効
成分とする微生物感染予防・治療剤に関する。従来、微生物の感染に対する予防あるいは治療
のための多くの薬物が知られているが、なお新規
な感染予防・治療剤の開発が望まれている。本発明者らは種々研究の結果、前記したごとき
PGまたはUDP―PGが優れた微生物感染予防・治
療の効果を有する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
抗腫瘍作用を有していることは知られているが、
本発明のPGおよびUDP―PGが微生物の感染に対
し予防あるいは治療の効果を示すことは本発明者
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。本発明の微生物感染予防・治療剤は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，p.1317〜1325，
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。本発明にかかるPGおよびUDP―PGは種々の微
生物の感染に対し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（感染予防効果）試験動物として約20ｇのddYマウス１群５匹を
使用した。PGの生理食塩水溶解液（８，２，0.5
mg／ml）0.5mlをマウス腹腔内に投与した。投与
量は各々200，50，12.5mg／Kgに相当する。投与
１日，４日，７日後に緑膿菌（Pseudomonas
aeruginosa BMH＃１）約10⁷個をマウス腹腔内
に攻撃した。攻撃７日後のマウスの生存数を第１
表に示す。対照群としてPG無投与群を設けた。 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-
This invention relates to a microbial infection prevention and treatment agent containing meso DAP (hereinafter abbreviated as UDP-PG) as an active ingredient. Although many drugs have been known to prevent or treat microbial infections, there is still a desire to develop new agents for preventing and treating infections. As a result of various studies, the present inventors found the above-mentioned
We have completed the present invention by discovering that PG or UDP-PG has excellent effects on preventing and treating microbial infections. It is known that analogues of the substances according to the present invention have adjuvant activity, have effects on T-lymph, B-lymph, complement system, pyrogenic leukocytes, and have antitumor activity. There are, but
This is the first finding by the present inventors that the PG and UDP-PG of the present invention exhibit preventive or therapeutic effects against microbial infection. The present invention will be explained in more detail below. The agent for preventing and treating microbial infection of the present invention is PG or
This is a composition containing UDP-PG as an active ingredient.
PG and UDP - PG is a biochemical and
Biophysical Research Communications (BBRC) vol.59, No.4, p.1317-1325,
It is a known substance reported in 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. It is found in 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. The PG and UDP-PG according to the present invention have preventive and therapeutic effects against infection by various microorganisms, and experimental examples regarding the pharmacological effects of these substances are shown below. Experimental Example 1 (Infection Prevention Effect) Groups of 5 ddY mice weighing approximately 20 g were used as test animals. PG dissolved in physiological saline (8, 2, 0.5
mg/ml) 0.5 ml was intraperitoneally administered to mice. The doses correspond to 200, 50, and 12.5 mg/Kg, respectively. Pseudomonas aeruginosa was detected 1, 4, and 7 days after administration.
aeruginosa BMH#1) were ^{intraperitoneally} challenged in mice. The number of surviving mice 7 days after challenge was determined as
Shown in the table. A group without PG administration was established as a control group.

【表】実験例２（感染予防効果）試験動物として約20ｇのddYマウス１群５匹を
使用した。PGの生理食塩水溶解液（８，２，0.5
mg／ml）0.5mlをマウスの腹腔内に投与した。投
与量は各々200，50，12.5mg／Kgに相当する。投
与１日後に大腸菌（Escherichia coli GN2411―
５）約10⁶個をマウス腹腔内に攻撃した。攻撃７
日後のマウスの生存数を第２表に示す。対照群と
してPG無投与群を設けた。[Table] Experimental Example 2 (Infection Prevention Effect) Groups of 5 ddY mice weighing approximately 20 g were used as test animals. PG dissolved in physiological saline (8, 2, 0.5
mg/ml) 0.5 ml was administered intraperitoneally to mice. The doses correspond to 200, 50, and 12.5 mg/Kg, respectively. One day after administration, Escherichia coli GN2411-
5) Approximately ¹⁰⁶ mice were intraperitoneally challenged. attack 7
The number of surviving mice after 1 day is shown in Table 2. A group without PG administration was established as a control group.

【表】実験例３（感染治療効果）試験動物として約20ｇのddYマウス１群５匹を
使用した。緑膿菌（Pseudomonas aeruginosa BMH
＃１）約10⁷個をマウス腹腔内に攻撃した。攻撃
直後、１時間後、２時間後にPGの生理食塩水溶
解液（0.4，０，２，0.05mg／ml）0.5mlを腹腔内
に投与した。投与量は各々10，５，1.25mg／Kgに
相当する。各々７日後の生存数を第３表に示す。
対照群としてPG無投与群を設けた。[Table] Experimental Example 3 (Infection Treatment Effect) Groups of 5 ddY mice weighing approximately 20 g were used as test animals. Pseudomonas aeruginosa BMH
#1) Approximately 10 ⁷ mice were intraperitoneally challenged. Immediately after the challenge, 1 hour later, and 2 hours later, 0.5 ml of a physiological saline solution of PG (0.4, 0, 2, 0.05 mg/ml) was administered intraperitoneally. The doses correspond to 10, 5, and 1.25 mg/Kg, respectively. Table 3 shows the number of survivors after 7 days.
A group without PG administration was established as a control group.

【表】実験例４試験菌として緑膿菌に替えて大腸菌
（Escherichia coli GN2411―５）を用い、PGの
投与を攻撃１時間後のみに行うほかは実験例３と
同様に行つて第４表に示す結果を得た。[Table] Experimental Example 4 The experiment was carried out in the same manner as in Experimental Example 3, except that Escherichia coli GN2411-5 was used instead of Pseudomonas aeruginosa as the test organism, and PG was administered only 1 hour after challenge. The results shown are obtained.

【表】実験例５（感染予防効果）試験薬物のPGに替えてUDP―PGを用いるほか
は実験例１と同様に行つて第５表に示す結果を得
た。[Table] Experimental Example 5 (Infection Prevention Effect) The same procedure as in Experimental Example 1 was performed except that UDP-PG was used instead of PG as the test drug, and the results shown in Table 5 were obtained.

【表】実験例６（感染予防効果）試験薬物のPGに替えてUDP―PGを用いるほか
は実験例２と同様に行つて第６表に示す結果を得
た。[Table] Experimental Example 6 (Infection Prevention Effect) The same procedure as in Experimental Example 2 was performed except that UDP-PG was used instead of PG as the test drug, and the results shown in Table 6 were obtained.

【表】実験例７（感染治療効果）試験薬物のPGに替えてUDP―PGを用いるほか
は実験例３と同様に行つて第７表に示す結果を得
た。[Table] Experimental Example 7 (Infection Treatment Effect) The same procedure as in Experimental Example 3 was performed except that UDP-PG was used instead of PG as the test drug, and the results shown in Table 7 were obtained.

【表】実験例８（感染治療効果）試験薬物のPGに替えてUDP―PGを用いるほか
は実験例４と同様に行つて第８表に示す結果を得
た。[Table] Experimental Example 8 (Infection Treatment Effect) The same procedure as in Experimental Example 4 was performed except that UDP-PG was used instead of PG as the test drug, and the results shown in Table 8 were obtained.

【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は微生物の感染に対する予防
および治療に優れた効果を示すことがわかる。ま
た本発明のPGおよびUDP―PGの効果は速効性
（１日前の投与で予防効果を示す）であり、この
効果はアジユバンド活性などの発現とは作用を異
にしているものと考えられる。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９に示
す。実験例９ 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。人の微生物感染症の治療または予防のために
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0.145〜23mg／Kg／dayが適
当である。以下に本発明にかかわる剤の具体例を実施例と
して示す。実施例１ PG3ｇ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 PG3ｇをリンゲル液100mlに溶かしてPGのリン
ゲル溶液をつくり、この10mlをアンプルに封入し
てPGの注射液とする。実施例３ PG3ｇを５％キシリツト液100mlに溶かしてPG
のキシリツト溶液をつくり、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 in preventing and treating microbial infections. Furthermore, the effects of PG and UDP-PG of the present invention are fast-acting (showing a preventive effect when administered one day before), and this effect is thought to be different from the expression of adjuvant activity. The results of measuring the acute toxicity (LD ₅₀ ) of PG and UDP-PG of the present invention are shown in Experimental Example 9 below. Experimental Example 9 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 and 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. When using the PG and UDP-PG of the present invention for the treatment or prevention of human microbial infections, PG
Alternatively, UDP-PG is mainly administered intravenously. For intravenous administration, it is used after being dissolved in physiological saline, Ringer's solution, 5% xylate, etc. The appropriate dosage is 0.145 to 23 mg/Kg/day. Specific examples of the agents related to the present invention are shown below as examples. Example 1 A PG saline solution was prepared by dissolving 3 g of PG in 100 ml of physiological saline, and 10 ml of this was sealed in an ampoule to prepare a PG injection solution. Example 2 A Ringer's solution of PG is prepared by dissolving 3 g of PG in 100 ml of Ringer's solution, and 10 ml of this is sealed in an ampoule to prepare a PG injection solution. Example 3 Dissolve 3g of PG in 100ml of 5% xyrite solution to make PG.
Prepare a xylitol 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 Microbial infection prevention using 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・Treatment agent.