JP2823717B2 - Method for producing stable antibacterial agent and antibacterial substance used therefor - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a stable antibacterial substance, 2-alkoxy (or aryloxy) -3,11.
The present invention relates to an antibacterial agent containing tridecadiene-5,7,9-triyn-1-ol and a method for producing the compound. The antibacterial agent of the present invention can be widely used as pharmaceuticals, agricultural chemicals, adhesives and the like.

[0002]

2. Description of the Related Art It is known that safflower contains a pigment having a high industrial value. However,
Since the safflower contains only a small amount of these pigments, a technique has been developed for producing a large amount of the pigment by tissue-culturing safflower calli.

It has recently been found that safflower contains not only pigments but also substances having other uses. For example, the following formula (II) present in safflower

[0004]

And formula (III)

[0006]

It has been found that the compound of the formula (1) has an antibacterial property (Agric. Biol. Chem., 41 (9), 1761 (1977)).
And Phytochemistry, 10 , 1579 (1971)). It is considered that these compounds can be mass-produced by callus culture of safflower as in the case of the pigment. However, the compounds of formulas (II) and (III) are not sufficiently stable, and their usefulness as antibacterial substances has been questioned. Moreover, studies on antibacterial substances derived from safflower are still shallow, and studies on antibacterial properties of compounds other than formulas (II) and (III) have not been made. Further, just because the compounds of formulas (II) and (III) have antibacterial properties, it is difficult to predict whether or not the compounds having similar structures have similar antibacterial properties. is there. For this reason, antibacterial substances that can be mass-produced by callus culture of safflower and have excellent stability have not yet been developed.

On the other hand, two of the compounds represented by the formula (I)
Compounds whose methoxy is methoxy are Phytochemistr
y, 18 , 829 (1979). However, this document has no description suggesting the antibacterial property of the compound.

[0009]

SUMMARY OF THE INVENTION An object of the present invention is to provide an antibacterial agent containing a stable antibacterial substance which can be mass-produced by liquid culture of safflower calli.

[0010]

Means for Solving the Problems] Such object is mainly anti
As bacterial active ingredient, the formula (I)

[0011]

Wherein R is an alkyl group, an alkenyl group,
This is achieved by the present invention which provides an antibacterial agent containing a compound represented by alkynyl group, cycloalkyl group, cycloalkenyl group or aryl group) and a method for producing the compound.

The substituent enumerated as R at the 2-position in the formula (I) is typically a commonly used substituent having 1 to 10 carbon atoms, but a substituent having 11 or more carbon atoms is used. You can also. Specifically, various substituents such as a methyl group, an ethyl group, a propyl group, and a butyl group can be used as the alkyl group. Further, the substituents listed as R may be further substituted with halogen or the like.

As is apparent from Example 1 described below, the compound of formula (I) was first shown to exhibit antibacterial activity against bacteria such as B. subtilis and E. coli . This compound is characterized in that it has higher stability than the compounds represented by the above formulas (II) and (III). Since the compound of the formula (III) has an epoxide structure, it easily undergoes a ring-opening reaction with an acid or an alkali to change to a diol, and the compound of the formula (II) reacts with an alkali to form another compound. It has the disadvantage of being easily changed. However, since the compound of the formula (I) used in the present invention does not have such a disadvantage and has a high affinity for a hydrophobic substance, the usefulness of the antibacterial agent containing the compound of the formula (I) is represented by the formula (II) And extremely high as compared with the compound of the formula (III).

The compound of the formula (I) can be contained in the antibacterial agent of the present invention at various concentrations depending on the purpose of use. Specifically, it can be used at 0.1-10%. Was
However, the compound of formula (I) is a major anti-microbial agent of the present invention.
It must be a bacterial active ingredient. This is a safflower child
Tissue culture of leaf-derived calli, simply treated with alcohol
The purpose is to exclude compositions that are only simple. That is,
Proportion of compound of formula (I) contained in such a composition
Only those that are positively enhanced are included in the antibacterial agent of the present invention.
included. Diluents include ethanol, methanol,
A volatile solvent such as acetone can be used. Antibacterial agents of the present invention, for example, pharmaceuticals such as colorants,
It is expected to be applied as an antifungal agent or adhesive applied to walls and the like. In addition, a wide variety of uses such as spraying as a disinfectant can be considered. For example, several g / 100 g can be mixed with lanolin or petrolatum.

Further, since the compound of the formula (I) can be synthesized in a large amount by the production method of the present invention,
The antibacterial agent of the present invention has extremely high industrial utility. The production method of the present invention comprises tissue culture of calli derived from safflower cotyledons and treatment with a substituted or unsubstituted alcohol or phenol. As the medium, for example, an MS medium or the like can be used. Treatment with a substituted or unsubstituted alcohol or phenol encompasses all operations which produce compounds of formula (I) by reaction with these substances. Usually, it is performed by adding these substances to the crude product of the culture and stirring. After treatment, the compound of formula (I) can be purified and isolated by a conventional method. For example, it can be extracted with ethyl acetate and further isolated by column chromatography.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the scope of the present invention should not be construed as being limited by such Examples.

[0018]

【Example】

(Example 1) According to the production method of the present invention, the compound represented by the formula (I)
The compound represented by was produced, and the property was examined.

A safflower callus was used as a cell group, and an MS medium (pH 5.7) having the composition shown in the following table, to which 2,4-D (10 ^-6 mol) was further added as a plant hormone, was used as a medium. Was.

[0020]MS medium Component amount (mg / l) NH_FourNO_Three 1650 KNO_Three 1900 KH_TwoPO_Four 170 CaCl_Two・ 2H_TwoO 440 MgSO_Four・ 7H_TwoO 370 MnSO_Four・ 4H_TwoO 22.3 H_ThreeBO_Four 6.2 ZnSO_Four・ 7H_TwoO 8.6 KI 0.83 NaMO_Four・ 2H_TwoO 0.25 CuSO_Four・ 5H_TwoO 0.025 CaCl_Two・ 6H_TwoO 0.025 FeSO_Four・ 7H_TwoO 27.8 Na_TwoEDTA 37.3 myo-inositol 100 4-amine HCl 0.1 pyridoxine HCl 0.5 nicotinic acid 0.5 glycine 2.0Saccharose 30.0  200 g (f, w) of safflower callus immersed and extracted with methanol
And fractionated with ethyl acetate. This ethyl acetate
The layer (190 mg) was treated as a methanol solution for 7 days
Was.

HPLC analysis (Inertsil ODS; acetonitrile-water as a developing solution) in the fractionated ethyl acetate layer was used to increase the concentration from 40% to 90% over 25 minutes; the flow rate was 0.8 ml / min. Temperature is room temperature; detection wavelength is 254
nm) The fraction with a tR value of 21.1 was isolated (yield 1.46 m).
g) The results of instrumental analysis were as follows.

¹ H NMR [in CDCl ₃ ]: δ 1.82
(3H), 3.33 (3H) 3.48 (1H), 3.59 (1H) 3.78 (1H), 5.54 (1H) 5.81 (1H), 6.19 (1H) 6 .40 (1H) UV [in MeOH] (nm): 260, 330, 350 MS: EI-MS mw: 214 From these analytical results, the compound of this fraction was found to be 2-methoxy-3,11-tridecadien-5. , 7,9-Triin-
It was found to be 1-ol. Even when the compound was stored in a methanol solution at room temperature for 7 days, almost no change was observed in the antibacterial activity.

(Embodiment 2) In this embodiment, the formula (I)
The antibacterial properties of the compound represented by are examined. Cotyledons 7 days after sowing were soaked and extracted with methanol for 7 days, and fractionated with ethyl acetate. The fractionated ethyl acetate layer was analyzed by HPLC under the same conditions as in Example 1. As a result, tR33.7 (fraction A), 27.7 (fraction B), 21.1 (fraction C: Example 1)
Compound 1) and peaks at 15.0 (fraction D) (Inertsil ODS; developing solution: 40-90% acetonitrile-water; flow rate: 0.8 ml / min; temperature: room temperature). The substances A, B, C and D were isolated from the fractionated ethyl acetate layer by a silica gel column (100-60% benzene-ethyl acetate). These substances were subjected to an antibacterial test by a paper disk method. When the test bacterium was B. subtilis and the diameter of the antibacterial zone by tetracycline as a control compound was +3, the activity of each component was +2 for A and D, and +1 for B and C. Also, E. col
When the same procedure was applied to i. , the same antibacterial activity was observed.

The UV spectra of A, B and D were measured to be 260 nm, 330 nm and 3 nm, respectively.
Absorption was observed at 50 nm.

──────────────────────────────────────────────────の Continuation of the front page (51) Int.Cl. ⁶ Identification code FI C12R 1:91) (C12P 7/22 C12R 1:91) (72) Inventor Akio Kobayashi 1-2-1, Tsushimanaka, Okayama City, Okayama Prefecture 2-103 (58) Field surveyed (Int. Cl. ⁶ , DB name) A01N 31/02 A01N 31/14 CA (STN) REGISTRY (STN)

Claims (2)

(57) [Claims] (1) As a main antibacterial active ingredient, a compound represented by the formula (I): Wherein R is an alkyl group, an alkenyl group, an alkynyl group, a cycloalkyl group, a cycloalkenyl group or an aryl group. , 9
-An antibacterial agent containing triin-1-ol. 2. A method comprising the steps of tissue-culturing callus derived from safflower cotyledons and treating with a substituted or unsubstituted alcohol or phenol. 2-alkoxy (or aryloxy) -3,11-tridecadien-5,7, A method for producing an antibacterial agent containing 9-triin-1-ol.