JP2754312B2 - Indene derivative and antibacterial agent containing the same as an active ingredient - Google Patents
Indene derivative and antibacterial agent containing the same as an active ingredientInfo
- Publication number
- JP2754312B2 JP2754312B2 JP9866993A JP9866993A JP2754312B2 JP 2754312 B2 JP2754312 B2 JP 2754312B2 JP 9866993 A JP9866993 A JP 9866993A JP 9866993 A JP9866993 A JP 9866993A JP 2754312 B2 JP2754312 B2 JP 2754312B2
- Authority
- JP
- Japan
- Prior art keywords
- indene derivative
- same
- antibacterial agent
- active ingredient
- agent containing
- 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
Description
【0001】[0001]
【産業上の利用分野】本発明はインデン誘導体及びこれ
を有効成分とする抗菌剤に関する。The present invention relates to an indene derivative and an antibacterial agent containing the same as an active ingredient.
【0002】[0002]
【従来の技術】ハトムギもやしの乾燥物、搾汁液又は抽
出物に抗菌活性のあることが知られており、かかる抗菌
活性を示す化合物としてステアリン酸又はパルミチン酸
のモノグリセライド、又はこれらの誘導体が示唆されて
いる(特開平2−270825、特開平3−24047
3)。2. Description of the Related Art It is known that a dried product, a squeezed liquid or an extract of barley sprouts has an antibacterial activity, and monoglycerides of stearic acid or palmitic acid or derivatives thereof are suggested as compounds exhibiting such antibacterial activity. (JP-A-2-270825, JP-A-3-24047)
3).
【0003】[0003]
【発明が解決しようとする課題】本発明は、ハトムギも
やしから単離され、優れた抗菌活性を示す、新規のイン
デン誘導体を提供するものである。SUMMARY OF THE INVENTION The present invention provides a novel indene derivative which is isolated from pearl sprouts and has excellent antibacterial activity.
【0004】[0004]
【課題を解決するための手段】しかして本発明者らは、
ハトムギもやしの乾燥物、搾汁液又は抽出物が示す抗菌
活性について鋭意研究した結果、ハトムギもやしに所定
の処理を施すと、新規のインデン誘導体が単離され、該
インデン誘導体が優れた抗菌活性を示すことを見出し
た。Means for Solving the Problems Thus, the present inventors have
As a result of diligent research on the antibacterial activity of the dried barley sprouts, the squeezed liquid or the extract, when a predetermined treatment is applied to the barley sprouts, a new indene derivative is isolated, and the indene derivative shows excellent antibacterial activity I found that.
【0005】すなわち本発明は、下記の式1で示される
インデン誘導体及びこれを有効成分とする抗菌剤に係
る。That is, the present invention relates to an indene derivative represented by the following formula 1 and an antibacterial agent containing the same as an active ingredient.
【0006】[0006]
【式1】 (Equation 1)
【0007】式1で示されるインデン誘導体はハトムギ
もやしを次のように処理することによって単離される。
先ず、詳しくは実施例で後述するように、ハトムギ種子
を暗培養してハトムギもやしを得る。対象となるハトム
ギ種子は、徳田在来種、中里在来種、岡山在来種、黒石
在来種等、その品種に特に制限はない。また暗培養は一
般のもやし暗培養条件にしたがうことができる。例え
ば、ハトムギ種子を25℃で4〜8日間程度暗培養すれ
ばよい。かくして得られるハトムギもやしはその全部位
を利用することができる。The indene derivative of the formula 1 is isolated by treating barley sprouts as follows.
First, as described in detail later in the Examples, the seeds of the barley are dark-cultured to obtain barley sprouts. There are no particular restrictions on the varieties of the target barley seeds, such as Tokuda native, Nakazato native, Okayama native, and Kuroishi native. In addition, dark culture can be performed according to general sprouts dark culture conditions. For example, the pearl barley seeds may be dark-cultured at 25 ° C. for about 4 to 8 days. The thus obtained barley sprouts can use all the parts.
【0008】次に、これも詳しくは実施例で後述するよ
うに、上記のハトムギもやし、その磨砕物、凍結乾燥
物、更には粉砕物等をメチルアルコールやエチルアルコ
ール等の低級アルコールで抽出処理し、その抽出物をク
ロロホルムやアセトン等で液液分配抽出処理して、アセ
トン易溶性の単純脂質を得る。[0008] Next, as will be described later in detail in the examples, the above-mentioned barley sprouts, their crushed, freeze-dried, and further crushed products are extracted with a lower alcohol such as methyl alcohol or ethyl alcohol. The extract is subjected to liquid-liquid partition extraction with chloroform, acetone or the like to obtain acetone-soluble simple lipids.
【0009】最後に、これもまた詳しくは実施例で後述
するように、上記の単純脂質を繰り返してクロマト分画
処理し、所望の化合物を単離する。クロマト分画処理
は、極性の異なる移動相を用い、ゲル浸透クロマトグラ
フィーと高速液体クロマトグラフィーとを組み合わせて
行うのが好ましい。Finally, as described later in detail in Examples, the above simple lipid is repeatedly subjected to chromatographic fractionation treatment to isolate a desired compound. The chromatographic fractionation treatment is preferably performed using a mobile phase having a different polarity, and a combination of gel permeation chromatography and high performance liquid chromatography.
【0010】かくして単離される化合物の構造解析結果
は下記の通りである。 (1)分子量:190(C11H10O3) (2)赤外線吸収スペクトル:1585,1703,3
050cm-1 (3)核磁気共鳴スペクトル(1H−NMR,δ):
3.99(3H,s),4.09(3H,s),5.9
8(1H,s),7.19(1H,dd,J=8.7
9,2.56),7.56(1H,d,J=2.5
6),8.00(1H,d,J=8.79) (4)核磁気共鳴スペクトル(13C−NMR,δ):5
7.7,58.7,105.6,112.3,118.
1,126.1,133.2,136.6,167.
2,169.3,181.5The results of the structural analysis of the compound thus isolated are as follows. (1) Molecular weight: 190 (C 11 H 10 O 3 ) (2) Infrared absorption spectrum: 1585, 1703, 3
050 cm -1 (3) Nuclear magnetic resonance spectrum ( 1 H-NMR, δ):
3.99 (3H, s), 4.09 (3H, s), 5.9
8 (1H, s), 7.19 (1H, dd, J = 8.7)
9, 2.56), 7.56 (1H, d, J = 2.5)
6), 8.00 (1H, d, J = 8.79) (4) Nuclear magnetic resonance spectrum ( 13 C-NMR, δ): 5
7.7, 58.7, 105.6, 112.3, 118.
1, 126.1, 133.2, 136.6, 167.
2,169.3,181.5
【0011】上記の構造解析結果から、単離される化合
物は式1で示されるインデン誘導体であり、3,5−ジ
メトキシ−1H−インデン−1−オンであることが決定
された。From the results of the above structural analysis, it was determined that the isolated compound was an indene derivative represented by the formula 1, and was 3,5-dimethoxy-1H-inden-1-one.
【0012】詳しくは実施例で後述するように、式1で
示されるインデン誘導体は、細菌、酵母、カビに対して
優れた抗菌活性を示し、天然抗菌剤として食品や化粧品
等への利用が注目される。As will be described later in detail in Examples, the indene derivative represented by Formula 1 exhibits excellent antibacterial activity against bacteria, yeasts and molds, and its use as a natural antibacterial agent in foods and cosmetics is attracting attention. Is done.
【0013】[0013]
試験区分1(インデン誘導体の単離) 収穫した徳田在来種のハトムギ種子を25℃で4日間暗
培養してハトムギもやしを得、該ハトムギもやしを磨砕
した後に棚温度20℃で凍結乾燥して、凍結乾燥物を得
た。Test Category 1 (Isolation of Indene Derivatives) The harvested barley seeds of the native Tokuda are cultivated in the dark at 25 ° C for 4 days to obtain barley sprouts. Thus, a freeze-dried product was obtained.
【0014】上記の凍結乾燥物150gにメチルアルコ
ール6リットルを加え、ホモジナイズ処理し、これを室
温で48時間放置した後、濾過して抽出液を得た。残渣
にメチルアルコール6リットルを加え、同様に抽出処理
を行なって抽出液を得、これを1回目の抽出液と合わせ
た。そして合わせた抽出液を減圧下に40〜45℃で加
熱してメチルアルコールを蒸発することにより抽出物3
0gを得た。6 liters of methyl alcohol was added to 150 g of the lyophilized product, homogenized, left at room temperature for 48 hours, and filtered to obtain an extract. 6 L of methyl alcohol was added to the residue, and an extraction treatment was performed in the same manner to obtain an extract, which was combined with the first extract. Then, the combined extract is heated at 40 to 45 ° C. under reduced pressure to evaporate methyl alcohol, thereby obtaining extract 3
0 g was obtained.
【0015】上記の抽出物30gにメチルアルコール1
00mlとクロロホルム200mlとを加え、混合撹拌し、
更に0.8%塩化カリウム水溶液60mlを加えて静置し
た。クロロホルム層を分取し、減圧下に40〜45℃で
加熱して1mlに濃縮した後、アセトン10mlと10%メ
チルアルコール性塩化マグネシウム溶液0.2mlとを加
え、混合撹拌し、1時間氷冷後、遠心分離して、上清の
アセトン溶液を分取した。残渣にアセトン10mlと10
%メチルアルコール性塩化マグネシウム溶液0.2mlと
を加え、同様に処理を行なって、アセトン溶液を得、こ
れを1回目のアセトン溶液と合わせた。そして合わせた
アセトン溶液を減圧下に40〜45℃で加熱してアセト
ンを蒸発することによりアセトン易溶性の単純脂質7.
1gを分離した。In 30 g of the above extract, methyl alcohol 1 was added.
00 ml and 200 ml of chloroform were added and mixed and stirred.
Further, 60 ml of a 0.8% aqueous potassium chloride solution was added, and the mixture was allowed to stand. The chloroform layer was separated, heated under reduced pressure at 40 to 45 ° C., concentrated to 1 ml, added with 10 ml of acetone and 0.2 ml of a 10% methyl alcoholic magnesium chloride solution, mixed and stirred, and cooled with ice for 1 hour. Thereafter, the mixture was centrifuged to separate an acetone solution of the supernatant. 10 ml of acetone and 10 ml
The mixture was treated in the same manner as above with 0.2 ml of a methyl alcoholic magnesium chloride solution to obtain an acetone solution, which was combined with the first acetone solution. 6. The combined acetone solution is heated at 40 to 45 ° C. under reduced pressure to evaporate the acetone, whereby the acetone is easily soluble simple lipid.
1 g was separated.
【0016】上記の単純脂質7.1gをクロロホルムで
溶解し、ジャイゲル 1H(JAIGEL 1H、商品
名、日本分析工業社製、8.0mmφ×500mm)を2本
用いてゲル浸透クロマトグラフィーを行なった。この
際、移動相としてクロロホルムを用い、該クロロホルム
を流速3.5ml/分で流下させた。RI(屈折率)で検
出し、主要なピークを示す4画分を得た。バチルス サ
ブチリス、サッカロミセス セレビシェ、アスペルギル
ス ニガーを検定菌として、各画分の抗菌試験をペーパ
ーディスク法により行ない、最も強い抗菌活性を示すR
t(保持時間、以下同じ)=55〜64分の画分1.8
gを得た。[0016] 7.1 g of the above simple lipid was dissolved in chloroform, and gel permeation chromatography was carried out using two pieces of JAIGEL 1H (trade name, manufactured by Nippon Kagaku Kogyo KK, 8.0 mmφ × 500 mm). At this time, chloroform was used as a mobile phase, and the chloroform was allowed to flow at a flow rate of 3.5 ml / min. Detected by RI (refractive index), four fractions showing major peaks were obtained. Using Bacillus subtilis, Saccharomyces cerevisiae and Aspergillus niger as test bacteria, an antibacterial test of each fraction was performed by a paper disc method, and R showing the strongest antibacterial activity was obtained.
t (retention time, the same applies hereinafter) = fraction of 55 to 64 minutes 1.8
g was obtained.
【0017】上記の画分1.8gをメチルアルコールで
溶解し、キャップセル パックC18(CAPCELL
PAK C18、商品名、資生堂社製、20mmφ×250
mm、以下同じ)を用いて高速液体クロマトグラフィーを
行なった。この際、移動相としてメチルアルコール/水
=20/80(重量比)で出発し、30分後に100/
0(重量比)となる直線グラジエントを行ない、該移動
相を流速3.0ml/分で流下させた。UV(紫外線吸光
度、254nm、以下同じ)で検出し、主要なピークを
示す6画分を得た。上記と同様の抗菌試験により、最も
強い抗菌活性を示すRt=34〜38分の画分146.
4mgを得た。1.8 g of the above fraction was dissolved in methyl alcohol, and the mixture was dissolved in Capcell Pack C 18 (CAPCELL).
PAK C18 , trade name, manufactured by Shiseido, 20mmφ × 250
mm, the same applies hereinafter) using high performance liquid chromatography. At this time, the reaction was started with methyl alcohol / water = 20/80 (weight ratio) as a mobile phase, and 100/30 minutes later.
A linear gradient of 0 (weight ratio) was performed, and the mobile phase was allowed to flow at a flow rate of 3.0 ml / min. Detected by UV (ultraviolet absorbance, 254 nm, hereinafter the same), 6 fractions showing major peaks were obtained. According to the same antibacterial test as described above, Rt = 34-38 min fraction showing the strongest antibacterial activity 146.
4 mg were obtained.
【0018】上記の画分146.4mgをメチルアルコー
ルで溶解し、キャップセル パックC18を用いて高速液
体クロマトグラフィーを行なった。この際、移動相とし
てメチルアルコール/水=50/50(重量比)を用
い、該移動相を流速3.0ml/分で流下させた。UVで
検出し、主要なピークを示す8画分を得た。上記と同様
の抗菌試験により、検定菌全てに強い抗菌活性を示すR
t=45分の化合物12.8mgを単離した。The above fraction (146.4 mg) was dissolved in methyl alcohol, and subjected to high performance liquid chromatography using Capcell Pack C18 . At this time, methyl alcohol / water = 50/50 (weight ratio) was used as the mobile phase, and the mobile phase was allowed to flow at a flow rate of 3.0 ml / min. Eight fractions detected by UV and showing major peaks were obtained. According to the same antibacterial test as above, R showing strong antibacterial activity against all test bacteria
12.8 mg of the compound at t = 45 min was isolated.
【0019】かくして単離した化合物の構造解析結果は
前述した通りであり、該化合物は式1で示される3,5
−ジメトキシ−1H−インデン−1−オンであった。The results of the structural analysis of the compound thus isolated are as described above.
-Dimethoxy-1H-inden-1-one.
【0020】試験区分2(インデン誘導体の抗菌活性) 抗菌活性は、バチルス サブチリス、サッカロミセス
セレビシェ、アスペルギルス ニガーを検定菌として、
ペーパーディスク法で試験した。バチルス サブチリス
は下記のトリプトソイ寒天培地(寒天1.5%)を、ま
たサッカロミセス セレビシェ及びアスペルギルス ニ
ガーは下記のポテトデキストロース寒天培地(寒天1.
5%)をそれぞれ使用し、これらの寒天培地15mlを径
90mmのシャーレに注入固化して、平板培地を作製し
た。 トリプトソイ寒天培地(日水製薬社製) ペプトン1.5%、ダイズペプトン0.5%、NaCl
0.5%及び寒天1.5% ポテトデキストロース寒天培地(日水製薬社製) ポテト浸出液末0.4%、ブドウ糖2.0%及び寒天
1.5%Test Category 2 (Antibacterial Activity of Indene Derivatives) The antibacterial activity was determined by Bacillus subtilis, Saccharomyces
Cerebiche, Aspergillus niger as test bacteria,
Tested by the paper disk method. Bacillus subtilis used the following tryptic soy agar medium (1.5% agar), and Saccharomyces cerevisiae and Aspergillus niger used the following potato dextrose agar medium (agar 1.
5%), and 15 ml of these agar media were injected into a 90 mm diameter petri dish and solidified to prepare a plate medium. Tryptic soy agar medium (Nissui Pharmaceutical Co., Ltd.) 1.5% peptone, 0.5% soy peptone, NaCl
0.5% and agar 1.5% Potato dextrose agar medium (manufactured by Nissui Pharmaceutical Co., Ltd.) 0.4% potato leachate powder, 2.0% glucose and 1.5% agar
【0021】検定菌を上記と同様の寒天培地15mlに接
種し、これを上記平板培地上に重層して固化した。重層
した平板培地上にペーパーディスク(東洋濾紙、径8m
m、薄手)を置き、該ペーパーディスクに単離した化合
物(10mg/ml)を20μl滴下し(200μg/ペーパ
ーディスク)、バチルス サブチリスは24時間培養
後、またサッカロミセス セレビシェ及びアスペルギル
ス ニガーは48時間培養後、生育阻止円の直径をそれ
ぞれ測定した。ペーパーディスク5枚の平均値をとり、
下記の式2により抗菌活性を求めた。The test bacteria were inoculated into 15 ml of the same agar medium as described above, and this was overlaid on the plate medium and solidified. Paper disk (Toyo filter paper, diameter 8m)
20 μl of the isolated compound (10 mg / ml) was dropped (200 μg / paper disc) on the paper disc, and Bacillus subtilis was cultured for 24 hours, and Saccharomyces cerevisiae and Aspergillus niger were cultured for 48 hours. And the diameter of the growth inhibition circle was measured. Take the average value of 5 paper discs,
The antibacterial activity was determined by the following equation 2.
【式2】抗菌活性(mm)=阻止円の直径(mm)−ペーパ
ーディスクの直径(8mm)[Formula 2] Antibacterial activity (mm) = Diameter of blocking circle (mm)-Diameter of paper disc (8 mm)
【0022】抗菌活性は、バチルス サブチリスに対し
て20.0mm、サッカロミセス セレビシェに対して1
1.5mm、アスペルギルス ニガーに対して11.5mm
であり、単離した化合物はこれらの細菌、酵母、カビに
対して優れた抗菌活性を示すことが検定された。The antibacterial activity is 20.0 mm against Bacillus subtilis and 1 against Saccharomyces cerevisiae.
1.5mm, 11.5mm for Aspergillus niger
The isolated compounds were tested to show excellent antibacterial activity against these bacteria, yeasts and molds.
【0023】[0023]
【発明の効果】既に明らかなように、以上説明した本発
明のインデン誘導体には抗菌剤として有効という効果が
ある。As is clear, the indene derivative of the present invention described above has an effect of being effective as an antibacterial agent.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 岡本 誉充 栃木県那須郡西那須野町西富山47番地 カゴメ那須寮315号 (72)発明者 坂本 秀樹 栃木県那須郡西那須野町大字井口字腰巻 47番地12 (58)調査した分野(Int.Cl.6,DB名) C07C 49/755 A01N 35/06 A23L 3/3544 A61K 31/12 ADZ CA(STN) REGISTRY(STN)──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor, Yoshimitsu Okamoto 47, Nishitoyama, Nishinasuno-cho, Nasu-gun, Tochigi Kagome-Nasu-Ryo 315 (72) Hideki Sakamoto 47, Kishimaki, Iguchi, Oguchi, Nisasu-cho, Nasu-gun, Tochigi 12 (58) Fields surveyed (Int.Cl. 6 , DB name) C07C 49/755 A01N 35/06 A23L 3/3544 A61K 31/12 ADZ CA (STN) REGISTRY (STN)
Claims (2)
分とする抗菌剤。2. An antibacterial agent comprising the indene derivative according to claim 1 as an active ingredient.
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP9866993A JP2754312B2 (en) | 1993-03-31 | 1993-03-31 | Indene derivative and antibacterial agent containing the same as an active ingredient |
DE69401151T DE69401151D1 (en) | 1993-03-31 | 1994-03-25 | Indene derivatives and antimicrobial agents containing these indene derivatives as active components |
EP94302155A EP0618182B1 (en) | 1993-03-31 | 1994-03-25 | Indene derivatives and antimicrobial agents containing them |
US08/372,729 US5534546A (en) | 1993-03-31 | 1995-01-17 | Indene derivatives and antimicrobial agents containing same |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP9866993A JP2754312B2 (en) | 1993-03-31 | 1993-03-31 | Indene derivative and antibacterial agent containing the same as an active ingredient |
Publications (2)
Publication Number | Publication Date |
---|---|
JPH06287158A JPH06287158A (en) | 1994-10-11 |
JP2754312B2 true JP2754312B2 (en) | 1998-05-20 |
Family
ID=14225927
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP9866993A Expired - Lifetime JP2754312B2 (en) | 1993-03-31 | 1993-03-31 | Indene derivative and antibacterial agent containing the same as an active ingredient |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP2754312B2 (en) |
-
1993
- 1993-03-31 JP JP9866993A patent/JP2754312B2/en not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
JPH06287158A (en) | 1994-10-11 |
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