Novel norbornene derivative and antifungal agent containing the same as active ingredient

Abstract

To identify an active ingredient in a mushroom extract having antifungal effect, and to provide a novel antifungal agent.SOLUTION: Provided is a norbornene derivative represented by the following formula (1) (where R1 represents H or OH, R2 represents H or a cyano group, and R3 represents CH3 or CHO), and an antifungal agent containing the norbornene derivative as an active ingredient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a novel borvonene derivative and an antifungal agent containing the same as an active ingredient. [Background technology]

[0002] Conventionally, anti-mold agents have been used when preserving items. For example, an anti-mold component has been proposed as an anti-mold agent for clothing, optical products, documents, books, paintings and calligraphy, and antiques, which comprises a container with an opening, a volatile anti-mold agent placed inside the container, and a gas-permeable film that closes the opening (Patent Document 1).

[0003] Antifungal agents are used especially for preserving textile products, fur products, dolls, etc. For example, it is described that antifungal agents such as o-phenylphenol (hereinafter sometimes referred to as "OPP"), p-chloro-m-xylenol, 3-methyl-4-isopropylphenol, allyl isothiocyanate, and octylisothiazoline are blended together with a volatile pyrethroid as an insect repellent component (Patent Document 2). In addition, Nn-butylcabamic acid-3-iodo-2-propynyl ester is described as a volatile antibacterial and antifungal agent (Patent Document 3).

[0004] On the other hand, it has also been disclosed that mushroom extracts are effective against certain bacteria. For example, an antibacterial agent containing, as an active ingredient, a water and / or organic solvent extract of one or more kinds of ascomycetes or basidiomycetes selected from the group consisting of Yamabushitake, Morel, Auricularia japonica, Bunashimeji, Nameko, Poria cordata, Usuhiratake, Sparassis crispa, etc., and an oral composition and food and drink containing the same have been disclosed (Patent Document 4). Also disclosed is a method for controlling plant pathogens, which comprises applying to plants a volatile substance emitted from the mycelium, fruiting body, or waste mushroom bed of one or more mushrooms selected from the group consisting of mushrooms of the genera Piptoporus, Mycoleptodonoides, Climacodon, Gloiothele, Lopharia, Microporus, Scytinostroma, Neolentinus, Hebeloma, Ischnoderma, Laetiporus, Daedaleopsis, Stecherinum, Trametes, Boidinia, Cymatoderma, Scopuloides, Ceriporia, and Pleurotus (Patent Document 5).

[0005] The present applicants have also discovered that solvent extracts of specific mushrooms are effective as volatile antifungal agents and have filed a patent application (Patent Document 6). [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Utility Model Application Publication No. 61-187303 [Patent Document 2] Japanese Patent Application Publication No. 11-139903 [Patent Document 3] Japanese Patent Application Publication No. 5-85909 [Patent Document 4] JP 2012-51897 A [Patent Document 5] JP 2011-167073 A [Patent Document 6] Patent application 2023-14974 Summary of the Invention [Problem to be solved by the invention]

[0007] An object of the present invention is to identify the active ingredient in the solvent extract of the specific mushroom described above, and to provide a novel antifungal agent that uses the active ingredient. [Means for solving the problem]

[0008] As a result of intensive research aimed at solving the above-mentioned problems, the inventors have discovered a group of compounds contained in a specific fraction of the solvent extract of the specific mushroom mentioned above, and further discovered that these compounds are active ingredients of an antifungal agent, thereby completing the present invention.

[0009] That is, the present invention relates to a compound represented by the following chemical formula (1): [ka] (In formula (1), R 1 is H or OH, R 2 is H or a cyano group, R 3 CH 3 or CHO) It is a borbonene derivative represented by the formula:

[0010] The present invention also relates to an antifungal agent comprising the above-mentioned borvonene derivative as an active ingredient. Effect of the Invention

[0011] The borvonene derivative of the present invention represented by chemical formula (1) is a novel compound and can be used for applications such as antifungal properties and fragrances.

[0012] The antifungal agent containing the borvonene derivative represented by the chemical formula (1) of the present invention as an active ingredient has a high antifungal effect against various types of mold.

[0013] Also, [Brief description of the drawings]

[0014] [Figure 1] FIG. 1 is a cross-sectional view of a petri dish used in an antifungal screening test. [Diagram 2] FIG. 2 is a plan view of a petri dish used in an antifungal screening test. [Diagram 3] FIG. 1 shows the results of LC-MS measurement of Compound 1. [Figure 4] FIG. 1 shows the results of measuring the 1H-NMR spectrum and the 13C-NMR spectrum of compound 1 (left, 1H-NMR spectrum; right, 13C-NMR spectrum). [Diagram 5] FIG. 1 shows NMR spectral assignments for Compound 1. [Figure 6] FIG. 1 shows the results of HH COSY (A) and HH NOESY (B) of compound 1, and the predicted three-dimensional structure (C). [Figure 7] FIG. 1 shows the results of LC-MS measurement of compound 2. [Figure 8] FIG. 1 shows the results of measuring 1H-NMR and 13C-NMR spectra of compound 2 (left, 1H-NMR spectrum; right, 13C-NMR spectrum). [Figure 9] FIG. 1 shows NMR spectral assignments for compound 2. [Figure 10] FIG. 1 shows the results of HH COSY (A) and HH NOESY (B) of compound 2, and the predicted three-dimensional structure (C). [Figure 11] FIG. 2 shows the results of CD spectrum measurement, optical rotation measurement, and UV absorption spectrum measurement of Compound 2. [Figure 12] FIG. 2 is a cross-sectional view of a petri dish used in an antifungal test. [Figure 13]FIG. 2 is a plan view of a petri dish used in an antifungal test. [Figure 14] FIG. 13 is a diagram showing a clothing cover of product example 3. [Figure 15] 4 shows the results of a confirmation test on the antifungal effect of the product 2 of the present invention. [Figure 16] 1 shows the results of a confirmation test on the antifungal effect of the product 1 of the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] The borvonene derivatives of the present invention (hereinafter sometimes simply referred to as "borvonene derivatives") are represented by the following chemical formula (1): These borvonene derivatives may be either α- or β-form, but α-form is preferred. [ka] In formula (1), R 1 is H or OH, R 2 is H or a cyano group, R 3 CH 3 or CHO. Among these borbonene derivatives, R 1 OH, R 2 is a cyano group, R 3 is the formula for CHO (2) [ka] Or, R 1 OH, R 2 is H, R 3 is the CHO formula (3) [ka] and more preferably, the compound represented by formula (3).

[0016] The borvonene derivative represented by the above formula (1) can be obtained by total synthesis according to a known method, or by further selecting a specific fraction from a solvent extract of Scutellaria of the genus Scutellaria of the family Agaricaceae as described in the specification of Japanese Patent Application No. 2023-14974 to obtain a compound represented by the following chemical formula (2) or (3), and further hydrogenating the CHO or cyano group while protecting other groups according to a known method as necessary.

[0017] A method for obtaining the borvonene derivative represented by the above chemical formula (2) or (3) by selecting a specific fraction from a solvent extract of Scutellaria Baicalensis will be described.

[0018] First, a strain of Scutellaria baicalensis grown on MMN medium (Marx 1969, Phytopathology 59:153-163.) is inoculated into MMN liquid medium and cultured with shaking at 25°C for 30 days. The culture filtrate is filtered with a suction filter to remove mycelium, and the resulting filtrate is extracted with ethyl acetate by a two-phase partition method. The resulting ethyl acetate partition layer is concentrated to dryness using an evaporator, and resuspended in 100% by mass (hereinafter simply referred to as "%) methanol to a concentration of 10 mg / 0.1 mL to obtain a culture solution.

[0019] The culture solution obtained above is fractionated by silica gel column chromatography. A mixed solvent of acetone and hexane is used as the developing solvent, and the acetone concentration is increased stepwise by 20% from 0% to 100%, and then eluted with methanol. By using 250 mL of solvent per fraction, seven fractions (0% fraction, 20% fraction, 40% fraction, 60% fraction, 80% fraction, 100% fraction, and methanol fraction) are obtained. Of these, the fraction with an acetone concentration of 20% is collected.

[0020] This 20% fraction is further fractionated by high performance liquid chromatography using the following conditions to obtain nine fractions (fraction 0 (retention time 0 - 3.8 minutes), fraction 1 (retention time 3.8 - 4.6 minutes), fraction 2 (retention time 4.6 - 6.8 minutes), fraction 3 (retention time 6.8 - 7.8 minutes), fraction 4 (retention time 7.8 - 9.4 minutes), fraction 5 (retention time 9.4 - 10.0 minutes), fraction 6 (retention time 10.0 - 10.8 minutes), fraction 7 (retention time 10.8 - 14.8 minutes), fraction 8 (retention time 14.8 - 16.0 minutes)). Among these, fraction 5 and fraction 6 are collected. <HPLC Conditions> Column: Cosmosil 5C18-AR-II 4.6ID×150mm T.FLOW: 0.8ml / min WAVE: 220nm / 280nm Solvent: ACN / millQ 5 - 100% (30min) Temperature: 40℃

[0021] From fraction 5 obtained in this way, a borvonene derivative represented by formula (2) is obtained, and from fraction 6, a borvonene derivative represented by formula (3) is obtained. Such fractions may be purified according to known methods.

[0022] Also, by appropriately drying the above fractions, the borvonene derivatives represented by formula (2) and formula (3) can be obtained as white solids.

[0023] Whether the above-mentioned borvonene derivative has been obtained can be determined by combining known measurement methods, such as NMR, 2D NMR, CD spectrum, optical rotation, UV absorption spectrum, etc. (For each measured value, refer to the examples).

[0024] The borvonene derivative thus obtained can be used as a fungicide because it has a fungicidal effect, and can also be used in fragrances, etc., similar to conventional β-borvonene, etc.

[0025] The case where the borvonene derivative of the present invention is used in an antifungal agent will be described. In this case, it is sufficient that the borvonene derivative of the present invention is contained as an active ingredient, so for example, one or more of each borvonene derivative, or one or two of fractions containing each borvonene derivative may be used as is. The content of the borvonene derivative in the antifungal agent of the present invention is not particularly limited, but is, for example, 0.1 to 100%, preferably 0.5 to 10%.

[0026] When a fraction containing each borvonene derivative is used as the active ingredient of the antifungal agent of the present invention, it can be concentrated, or this concentrated fraction can be dispersed in a suitable solvent as necessary to form a liquid agent. It can also be supported on a carrier to form a solid agent. When a solid of each borvonene derivative is used as the active ingredient of the antifungal agent of the present invention, it can be dispersed in a suitable solvent as necessary to form a liquid agent, or it can be directly or supported on a carrier to form a solid agent, just like the case of using the above fraction.

[0027] The solvent for dispersing the borbonene derivative may be water, methanol, ethanol, butanol, hexane, diethyl ether, ethyl acetate, acetone, acetonitrile, dichloromethane or a mixture thereof. Of these, it is preferable to use methanol.

[0028] The carrier for supporting the borbonene derivative is not particularly limited, but examples thereof include wood, cellulose, paper, rayon, polyethylene, polypropylene, polyester, polyurethane, wool, talc, clay, unglazed pottery, cloth, nonwoven fabric, silica, talc, activated carbon, silica gel, zeolite, cellulose beads, activated carbon, ceramics, and the like. Also included are sublimable carriers such as triisopropyltrioxane and cyclododecane.

[0029] In addition, since the borbonene derivatives are volatile, they can be mixed or used in combination with other volatile agents. Here, volatility refers to the property of easily volatilizing into the atmosphere at normal temperature and pressure. Other volatile agents include insect repellents, fragrances, deodorants, harmful substance removers, and freshness-preserving agents for fruits and vegetables. By mixing with an insect repellent, an insect repellent and antifungal agent can be provided. Furthermore, by mixing with a fragrance, an antifungal aromatic can be provided.

[0030] The above-mentioned insect repellents include pyrethroid compounds such as paradichlorobenzene, naphthalene, camphor, 2-phenoxyethanol, empentrin, fenothrin, allethrin, transfluthrin, profluthrin, and metofluthrin.

[0031] Examples of the aromatic components include animal-based fragrances such as musk, spirit cat fragrance, and dragon jasmine fragrance, and vegetable-based fragrances such as abies oil, acorn oil, almond oil, angelica root oil, peper oil, bergamot oil, perch oil, boa bur rose oil, kayabuchi oil, gananga oil, capsicum oil, caraway oil, cardamom oil, cassia oil, celery oil, cinnamon oil, citronella oil, cognac oil, coriander oil, cumin oil, camphor oil, dill oil, estugoran oil, eucalyptus oil, fennel oil, garlic oil, ginger oil, grapefruit oil, hop oil, lemon oil, lemongrass oil, nutmeg oil, mandarin oil, peppermint oil, pine oil, orange oil, sage oil, star anise oil, and turpentine oil. As the fragrance, artificial fragrances such as synthetic fragrances or extracted fragrances can also be used, and examples thereof include hydrocarbon fragrances such as pinene and limonene, alcohol fragrances such as linalool, geraniol, citronellol, menthol, borneol, benzyl alcohol, anise alcohol, and β-phenethyl alcohol, phenol fragrances such as anethole and eugenol, aldehyde fragrances such as n-butyraldehyde, isobutyraldehyde, hexylaldehyde, citral, citronellal, benzaldehyde, cinnamic aldehyde, and cumin aldehyde, ketone fragrances such as carvone, menthone, camphor, acetophenone, and ionone, lactone fragrances such as γ-butyrolactone, coumarin, and cineol, and ester fragrances such as octyl acetate, benzyl acetate, cinnamyl acetate, butyl propionate, and methyl benzoate. Furthermore, a blended flavor may be used which is a mixture of two or more of the above flavors.

[0032] Furthermore, the borbonene derivatives can be used in combination with other conventionally known volatile antifungal agents.

[0033] Examples of such conventionally known volatile antifungal agents include phenolic compounds such as allyl isothiocyanate, thymol, parachlorometaxylenol, and orthophenylphenol, 3-iodo-2-propynyl butylcarbamate, 3-methyl-4-isopropylphenol, N-(fluorodichloromethylthio)-phthalimide, and / or N-dichlorofluoromethylthio-N',N'-dimethyl-N-phenylsulfamide.

[0034] The fungicide containing the borvonene derivative of the present invention as an active ingredient described above can be applied to fungi to inhibit the growth of the fungi, but since the borvonene derivative is preferably volatile, the fungicide is preferably volatilized in air and applied to the fungi to inhibit the growth of the fungi. In this case, the fungicide of the present invention can exert its fungicidal effect by volatilizing about 10 mg as the fungicide. In addition, as the active ingredient of the fungicide, the borvonene derivative represented by formula (2) or formula (3) is preferred from the viewpoint of effectiveness, and the borvonene derivative represented by formula (3) is particularly preferred.

[0035] In volatilizing the antifungal agent of the present invention, for example, the material is mixed with a material constituting a sheet-like material such as a nonwoven fabric, and the material and the material are both formed into a sheet, or a suspension containing the material is applied to a sheet-like material such as a nonwoven fabric, and the liquid is evaporated as necessary, thereby supporting the material on an absorbent core, etc. In addition, in volatilizing the volatile antifungal agent of the present invention, for example, a method of volatilizing using a suitable volatilizing device, or a method of volatilizing in an atomized state using an atomizing device such as a pump spray, an aerosol, an ultrasonic vibrator, a pressurized liquid spray spray, or a pressurized air atomizing spray device, etc., can be mentioned, and these methods allow the agent to volatilize in an ordinary living space.

[0036] When the material is formed into a sheet-like product, a particular example is a wet-laid nonwoven fabric formed by weaving the material together with fibers, which allows the material to be supported relatively easily on the nonwoven fabric.

[0037] Methods for applying the antifungal agent of the present invention to a sheet-like material include known coating methods such as a dip coater method, a spray coater method, a flow coater method, a die coater method, a roll coater method, a blade coater method, a rod coater method, a bar coater method, and a spin coater method.

[0038] The antifungal agent of the present invention can be applied to nonwoven fabric by printing. Printing does not require any special printing equipment or processing equipment, and existing printing machines such as screen printing, gravure printing, lithographic printing, and inkjet printing can be used as they are.

[0039] The antifungal agent of the present invention can be volatilized in space to prevent mold from growing on stored objects, etc. For example, it can be volatilized in a chest of drawers, closet, clothing case, accessory case, camera case, etc. to prevent mold from growing on stored items such as clothes, leather products, and precision instruments such as cameras.

[0040] Furthermore, by volatilizing the antifungal agent of the present invention in a toilet or bathroom, it is possible to prevent mold on the hand basin, toilet bowl, floor, bathroom floor, walls, and bathtub.

[0041] In addition, by volatilizing the anti-fungal agent of the present invention inside a food storage facility, it is possible to prevent mold growth in foods such as citrus fruits, apples, leafy vegetables such as cabbage and Chinese cabbage, dairy products, grains such as wheat flour, bread, and rice cakes.

[0042] Furthermore, the antifungal agent of the present invention can be evaporated into a living space to prevent mold on bedding, sofas, walls, floors, tatami mats, etc.

[0043] Furthermore, by producing a clothing cover using a nonwoven fabric or the like treated with the antifungal agent of the present invention, a clothing cover having antifungal properties can be manufactured.

[0044] The types of mold whose growth can be inhibited by the antifungal agent of the present invention are not particularly limited, but examples include one or more types of mold belonging to the genera Cladosporium, Penicillium, Aspergillus, and Alternaria, preferably one or more types of mold belonging to the genera Cladosporium, Penicillium, and Aspergillus, and more preferably all types. Among these molds, the growth of one or more of Cladosporium sphaerospermum, Penicillium citrinum, Aspergillus niger, and Alternaria brassicicola can be preferably inhibited, and preferably one or more of Cladosporium sphaerospermum, Penicillium citrinum, and Aspergillus niger, and more preferably all of them can be inhibited.

[0045] The antifungal agent of the present invention can also be used for various antifungal applications, such as antifungal applications on walls and hard surfaces of bathrooms, washrooms and toilets, and on clothes stored in humid closets and the like; antifungal applications on citrus fruits such as mandarin oranges, apples, leafy vegetables such as cabbage and Chinese cabbage, grains such as wheat flour, dairy products, and in living spaces; and antifungal applications on bread, rice cakes, etc. EXAMPLES

[0046] The present invention will be described in detail below by way of examples of the present invention, but the present invention is not limited to these examples in any way.

[0047] Manufacturing Example 1 Fractionation of solvent extract of Scutellaria Baicalensis: Using Scutellaria baicalensis, a mushroom belonging to the genus Scutellaria in the family Agaricaceae, a solvent extract of Scutellaria baicalensis was obtained by the following method.

[0048] (Preparation of culture medium for Scutellaria Baicalensis) The strain of Scutellaria baicalensis grown on MMN medium was inoculated into MMN liquid medium and cultured with shaking at 25°C for 30 days. The culture filtrate was filtered with a suction filter to remove mycelium, and the obtained filtrate was partitioned and extracted with ethyl acetate. The obtained ethyl acetate partitioned layer was concentrated to dryness using an evaporator and resuspended in 100% methanol to a concentration of 10 mg / 0.1 mL to obtain a culture solution.

[0049] (Preparation of acetone fraction) The culture solution obtained above was fractionated by silica gel column chromatography. A mixed solvent of acetone and hexane was used as the developing solvent, and the acetone concentration was increased stepwise by 20% from 0% to 100%, and then eluted with methanol. 250 mL of solvent was used per fraction. As a result, seven fractions were obtained, namely, a 0% fraction (0.4 mg), a 20% fraction (26.0 mg), a 40% fraction (15.0 mg), a 60% fraction (7.6 mg), an 80% fraction (3.9 mg), a 100% fraction (15.9 mg), and a methanol fraction (3.3 mg) in terms of acetone concentration.

[0050] Example 1 Antifungal screening test: The seven fractions were subjected to antifungal tests using Penicillium citrinum NBRC6352 (hereafter referred to as PC).

[0051] (1) Preparation of PC spore suspension PC was inoculated onto potato dextrose agar medium (Difco Potato Dextrose Agar 39 g, distilled water 1 L) in a petri dish and cultured at 25°C for 2 to 4 weeks. The spores formed on the medium were then suspended in sterilized water with a paintbrush, and the suspension was centrifuged to precipitate the spores. The supernatant was discarded, and the precipitated spores were suspended in 10% potato dextrose liquid medium. The spore concentration of the suspension was then measured using a hemocytometer to determine a concentration of 5.0 × 10 5 The concentration was adjusted to be 1 / ml.

[0052]

number

[0053] Figures 1 and 2 show a cross-sectional view and a plan view of a petri dish used in the anti-mold screening test. The petri dish 1 was placed with the lid side down, a Kimwipe 2 containing water was placed on the lid side, and a slide glass 4 with a PC spore suspension 3 dropped thereon was placed. On top of that, each of the above fractions adjusted to be dissolved in hexane to a concentration of 5 mg / mL and impregnated into filter paper 5 with 1 mL was pressed with tweezers so as not to fall to the bottom side and set on top of the lid side, covered with a paraffin film around the petri dish and sealed, and cultured at 25 °C for 24 hours. After culturing, 0.002 ml of lactophenol blue solution was dropped to stain the hyphae and spores. The slide glass was covered, and the number of spores and the number of germinated spores were counted at a magnification of 100 times (eyepiece lens 10 times, objective lens 10 times) using a microscope, and the germination inhibition rate was determined from the following calculation formula. Also, the hyphal elongation inhibition rate was determined from the following calculation formula. The results are shown in Table 1.

[0054]

Equation

[0055]

Equation

[0056]

Table 1

[0057] As a result of the above activity test, the 20% fraction had a large amount (26.0 mg) and showed high activity.

[0058] The highly active 20% fraction (26.0 mg) was fractionated by high performance liquid chromatography (LC20A: manufactured by Shimadzu Corporation) at the retention times shown in Table 2 below. Table 2 also shows the fraction masses. <HPLC Conditions> Column:Cosmosil 5C18-AR-II4.6ID×150mm T.FLOW:0.8ml / min WAVE:220nm / 280nm Solvent:ACN / millQ 5-100%(30min) Temperature: 40℃

[0059] [Table 2]

[0060] The same antifungal screening test was carried out for each fraction. The results are shown in Table 3. As a result, fractions 5 and 6 showed high activity. The structures of compound 1 contained in fraction 5 and compound 2 contained in fraction 6 were determined.

[0061] [Table 3]

[0062] Example 2 Structure determination of compound 1: (Structure determination) The results of LC-MS for compound 1 contained in fraction 5 are shown in Figure 3. From these results, compound 1 was estimated to be a compound with a molecular weight of 259. In addition, high-resolution MS revealed that the molecular formula was C 16 H 21 NO 2 It was found that the compound had an unsaturation degree of 7 (results not shown). Compound 1 was further dissolved in deuterated chloroform, and the 1H-NMR spectrum (600 MHz) 13 C-NMR spectrum (150 MHz) was measured using a JEOL JNM-ECZ600 (JEOL). 1 H-NMR spectrum, 13 The C-NMR spectrum is shown in Figure 4. The NMR spectrum assignments are shown in Figure 5. From this, it was estimated that compound 1 has an α-borbonene skeleton. 1 H- 1H. COSY, 1 H- 1 H NOESY, HMBC, and HMQC spectra were also measured. 1 H- 1 H. COSY, 1 H- 1 H NOESY is shown in Figure 6. The three-dimensional structure predicted from this is also shown in Figure 6. Taking all of this into consideration, compound 1 obtained in fraction 5 was identified as being represented by the following chemical formula (2). (Hereinafter, this will be referred to as product 1 of the present invention.) When fraction 5 was dried, a white solid was obtained.

[0063] [ka]

[0064] Example 3 Structure determination of compound 2: The results of LC-MS for compound 2 contained in fraction 6 are shown in Figure 7. From these results, compound 2 was estimated to be a compound with a molecular weight of 234. In addition, high-resolution MS revealed that the molecular formula was C 15 H 22 O 2 Compound 1 was found to be a compound with an unsaturation degree of 5 (results not shown). Compound 1 was further dissolved in deuterated chloroform, 1 H-NMR spectrum (600MHz), 13 C-NMR spectrum (150 MHz) was measured using a JEOL JNM-ECZ600 (JEOL). 1 H-NMR spectrum and 13 The C-NMR spectrum is shown in Figure 8. The NMR spectrum assignments are shown in Figure 9. From this, it was estimated that compound 1 has an α-borbonene skeleton. 1 H- 1 H. COSY, 1 H- 1 H NOESY, HMQC, and HMBC spectra were also measured. 1 H- 1 H. COSY, 1 H- 1H NOESY is shown in Figure 10. The three-dimensional structure predicted from this is also shown in Figure 10. Furthermore, CD spectrum measurement, optical rotation measurement, and UV absorption spectrum measurement were performed. These results are shown in Figure 11. Taking these into consideration, compound 2 obtained in fraction 6 was identified as being represented by the following chemical formula (3). (Hereinafter, this will be referred to as product 2 of the present invention.) When fraction 6 was dried, a white solid was obtained.

[0065] [ka]

[0066] Example 4 Anti-mold test: A test was carried out to confirm the antifungal effect of invention product 2.

[0067] (1) Antifungal test Antifungal tests were carried out on the following mold spores. (1) Cladosporium sphaerospermum NBRC6348 (hereafter abbreviated as CS) (2) Penicillium citrinum NBRC6352 (hereinafter abbreviated as PC) (3) Aspergillus niger NBRC9455 (hereinafter abbreviated as AN)

[0068] (Preparation of collodion glass) A solution of collodion (5%):diethyl ether = 1:1 was prepared, and a cleaned slide glass (25 mm x 75 mm) was immersed in the above solution for 1 minute. The slide glass was removed from the solution and dried for 1 minute to prepare a collodion glass. (Preparation of Potato Dextrose (PDB) Liquid Medium for Suspending Mold Spores) Potato dextrose (PDB) liquid medium for suspending each mold spore was adjusted to the following concentrations using sterilized water. For CS: PDB concentration 1% For PC: PDB concentration 40% For AN: PDB concentration 1.3%

[0069] (Preparation of mold spore suspension) 0.1 ml of a glycerin solution (20% glycerin, 80% water) of each mold was smeared on potato dextrose agar medium and cultured at 25°C for 14 days. After that, about 7 cm of the mold was scraped off with a platinum loop and suspended in each of the potato dextrose liquid media prepared above. Next, the suspension was passed through a filter tip to remove the mycelium, and each mold suspension was obtained. The number of spores was measured at five points using a hemocytometer, the average was calculated, and the spore number was calculated using the same formula as in Example 1. The final number was 5.0 x 10 5 The cells were diluted to about 1 / ml.

[0070] 12 and 13 show the cross-sectional view and the plan view of the petri dish used in the antifungal test. A filter paper 7 moistened with 1 ml of sterile water was placed on the lid side of a high-waisted petri dish 6 with a diameter of 90 mm and a height of 200 mm, and a collodion glass 8 was placed on top of the filter paper 7, and 0.01 ml of each spore suspension was dripped onto the collodion glass in three dots (9). Meanwhile, a filter paper 10 was pressed with tweezers to the bottom side of the high-waisted petri dish so as not to fall, and 0.1 ml of the product 2 of the present invention (fraction 6) was dripped evenly. Immediately after that, 0.9 ml of hexane was dripped and dried at room temperature for 2 minutes. After drying, the bottom side was placed over the lid side, the petri dish was wrapped around the periphery of the petri dish with parafilm to seal it, and the dish was cultured at 25°C for 24 hours. After the culture, 0.002 ml of lactophenol blue solution was dripped to stain the mycelium and spores. The number of spores and germinated spores was counted at 100x magnification (10x eyepiece, 10x objective) using a microscope, and the average value of the three locations was calculated. The germination inhibition rate was calculated using the same formula as in Example 1. As a blank, 0.1 ml of methanol was dropped instead of the product 2 of the present invention. The results are shown in Table 4.

[0071] [Table 4]

[0072] As is clear from Table 4, the product 2 of the present invention exhibited excellent antifungal effects against all of CS, PC, and AN.

[0073] Manufacturing Example 2 <Preparation of anti-mold sheet> Weight 15g / m 2 The above-obtained invention product 1 (fraction 5) was applied by solid printing using gravure printing onto a SMMS nonwoven fabric (made of PP) (manufactured by Asahi Kasei Corporation).The nonwoven fabric was then dried to obtain an antifungal sheet.

[0074] Manufacturing Example 3 <Making a clothing cover for a hanger> A polypropylene film (thickness 0.025 mm) and the antifungal sheet obtained in Production Example 2 were overlapped and cut using ultrasound into the shape of the front sheet 11 and back sheet 12 shown in Figure 14, and at the same time, both sheets were bonded at the upper edge 14 and the side edge 15, excluding the neck part 13 and the lower part 16, to produce a garment cover 20 with antifungal properties. This garment cover is hung on a hanger 17 for use.

[0075] Example 5 Anti-mold test: A test was carried out to confirm the antifungal effect of invention product 2.

[0076] The mold spores were prepared by using Alternaria brassicicola O-264 (stock stock of Tottori University) (hereinafter referred to as AB) and the final concentration was adjusted to 10 ppm. The same procedure as in the mold prevention test of Example 4 was repeated to determine the germination inhibition rate. The results are shown in Table 5.

[0077] [Table 5]

[0078] As is clear from Table 5, the product 2 of the present invention also exhibited excellent antifungal effect against AB.

[0079] Example 6 Anti-mold test: A test was carried out to confirm the antifungal effect of invention product 2.

[0080] Except for using AN and CS together with PC as the mold spores, the hyphal growth inhibition rate was determined in the same manner as in the antifungal screening test of Example 1. The results are shown in Table 6.

[0081] [Table 6]

[0082] As is clear from Table 6, invention product 2 exhibited excellent antifungal effects against all of CS, PC, and AN.

[0083] Example 7 Anti-mold test: A test was carried out to confirm the antifungal effect of invention product 2.

[0084] 1 and 2 show a cross-sectional view and a plan view of the petri dish used in the antifungal screening test. The petri dish 1 was placed with the lid side facing down, a water-soaked Kimwipe 2 was laid on the lid side, and a slide glass 4 on which a PC spore suspension 3 was dropped was placed. On top of the petri dish, 10 μl of each fraction diluted with methanol was dissolved in 1 ml of hexane, and the filter paper was soaked in such a way that the air concentration would be 10 ppm if all the compounds in the solution were evaporated. The filter paper was pressed with tweezers to prevent it from falling to the bottom side, and the petri dish was covered with parafilm to seal it, and cultured at 25 ° C. for 24 hours. The product 2 of the present invention was adjusted so that the air concentration would be 0, 1.0 ppm, 2.5 ppm, 5.0 ppm, and 10.0 ppm if all the compounds in the solution were evaporated. After the culture, 0.002 ml of lactophenol blue solution was dropped to stain the mycelium and spores. A glass slide was placed over the specimen, and the length of the hyphae was measured using a microscope at 100x magnification (10x eyepiece, 10x objective). The results are shown in Figure 15.

[0085] As is clear from FIG. 15, invention product 2 exhibited a concentration-dependent antifungal effect.

[0086] Example 8 Anti-mold test: A test was carried out to confirm the antifungal effect of the product 1 of the present invention.

[0087] 1 and 2 show a cross-sectional view and a plan view of the petri dish used in the antifungal screening test. The petri dish 1 was placed with the lid side facing down, a water-soaked Kimwipe 2 was laid on the lid side, and a slide glass 4 on which a PC spore suspension 3 was dropped was placed. On top of the petri dish, 10 μl of each fraction diluted with methanol was dissolved in 1 ml of hexane, and the filter paper was soaked in such a way that the air concentration would be 10 ppm if all the compounds in the solution were evaporated. The filter paper was pressed with tweezers to prevent it from falling to the bottom side, and the petri dish was covered with parafilm to seal it, and cultured at 25 ° C. for 24 hours. The product 1 of the present invention was adjusted so that the air concentration would be 0, 0.25 ppm, 0.5 ppm, 1 ppm, and 2.5 ppm if all the compounds in the solution were evaporated. After the culture, 0.002 ml of lactophenol blue solution was dropped to stain the mycelium and spores. The number of spores and germinated spores were counted under a microscope at 100x magnification (10x eyepiece, 10x objective lens) to determine the germination rate. The results are shown in Figure 16.

[0088] As is clear from FIG. 16, the product 1 of the present invention exhibited a concentration-dependent antifungal effect. [Industrial Applicability]

[0089] The borvonene derivative of the present invention represented by chemical formula (1) is a novel compound and can be used for applications such as antifungal properties and fragrances. [Explanation of symbols]

[0090] 1 Petri dish 11 Front sheet 2 Kimwipes 12 Back sheet 3 PC spore suspension 13 Neck 4 Slide glass 14 Upper edge 5 Filter paper (soaked in extract) 15 Side edge 6 Waist-high Petri dish 16 Lower part 7 Moist filter paper 17 Hanger 8 Collodion glass 20 Clothing cover 9. Spore suspension 10 Filter paper (soaked in extract)

Claims

1. The following chemical formula (1) 【Chemistry 1】 (In formula (1), R １ is H or OH, R ２ is H or a cyano group, R ３ is CH ３ or CHO) A borbonene derivative represented by the formula:

2. 2. A fungicide comprising the borvonene derivative according to claim 1 as an active ingredient.

3. 3. The antifungal agent according to claim 2, wherein the fungus is one or more kinds of fungi belonging to the genera Cladosporium, Penicillium, Aspergillus and Alternaria.

4. 3. The antifungal agent according to claim 2, wherein the fungus is one or more species selected from the group consisting of Cladosporium sphaerospermum, Penicillium citrinum, Aspergillus niger and Alternaria brassicicola.

5. The antifungal agent according to claim 2, which is volatile.

6. A method for preventing mold, comprising applying the antifungal agent according to any one of claims 2 to 5 to mold to inhibit the growth of the mold.

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