CN1206331A - Fungicidal compounds - Google Patents

Abstract

Use of dunnione of formula (I) as a fungicide is provided. Fungicidal compositions containing dunnione are also provided.

Description

Fungicidal compounds

The present invention relates to dunnione as the application of bactericide and the preparation of compositions method that contains the dunnione that is fit to this application.

The ability that fungi produces resistance to bactericide means that the new bactericide of continuous needs prevents and treats them.Known some naphthoquinones and derivative thereof have bactericidal activity, as US4970328 and US4929642.Jacobsen and Wengel (Pestic.Sci., 1986,17,686-690) prepared on the carbon of furans oxygen ortho position and had 1 of one or two alkyl substituent, the furan derivatives of 4-naphthoquinones (A) and (B), and makes further research them.Find that dialkyl derivatives (B) has medium bactericidal activity, monoalkyl derivative (A) is non-activity then; Obviously, the bactericidal activity of these two compounds is not so good as some non-furan compounds that Jacobsen and wengel studied.

The inventor has now found that naturally occurring naphthoquinones sterilization derivative dunnione can be used as bactericide, and they have superior bactericidal activity, and especially the bacterial strain that present used bactericide is had resistance is all the more so.Other advantage of natural bactericidal agent comprises natural products source and potential security products.

A first aspect of the present invention has provided the application of dunnione as bactericide.The structure of dunnione is seen shown in the formula I.It is also understood that because dunnione can exist with different enantiomeric forms, thereby the present invention not only comprises these single enantiomers but also comprises the mixture of these enantiomers.Dunnione can extract from the acrial part of streptocarpus plant (Streptocarpus dunnii) or slipperwort platymiscium (Calceolaria Integrifolia) and obtain, perhaps prepared by II or III by the acid catalysis ring-closure reaction, III wherein is to utilize the aqueous solution of methyl alcohol and potassium hydroxide to be formed by IV.

Second aspect of the present invention provided bactericidal composition, and said composition comprises carrier and as the dunnione of active component.This must also comprise this method for compositions of preparation on the one hand, is about to dunnione and mixes with at least a carrier.Equally, it is contemplated that the different enantiomers or the mixture of enantiomers of dunnione may have different activity levels or action spectrum, thereby the present composition may comprise single enantiomer or mixture of enantiomers.

The present composition can contain 0.0001-0.1% (weight) formula I active component.When composition for preceding mixed matching the time, preferably they contain 0.0002-0.075% (weight) active component.Yet composition also can be sold with high concentration (for example high to 95%) the concentrate form of preceding dilution.

For the preparation present composition, dunnione can be mixed with various suitable inert carriers such as solvent, thinner and/or surfactant, form pulvis, granular solids, but warm nature pulvis or other solid pharmaceutical preparation or emulsion, missible oil, spray, aerosol or other liquid preparation.The suitable solvent and thinner comprise water, and aliphatic series or aromatic hydrocarbon such as dimethylbenzene or other petroleum distillate and alcohol are as ethanol.Surfactant can be anion, cation or nonionic.Wherein can also comprise antioxidant or other stabilizing agent and spices and colouring agent.Used identical in the type of these inert carriers and amount ratio and the conventional sterilization composition.

Except that these inert carriers, can also comprise one or more other active components in the present composition.These other active components both can be for having had other compound of bactericidal activity, also can be for The compounds of this invention being demonstrated other compound of synergistic effect.

The 3rd aspect of the present invention provides the method for kill harmful fungi, and this method comprises the place with dunnione recited above or the described harmful fungoid invasion and attack of compositions-treated.Preferably, above-mentioned place comprises the environment that fungi itself or these fungies will attack or attack.

The present invention further specifies with the following example.Embodiment 1-prepares dunnione

Cooke described the preparation [nature, (1948), 163,178] of dunnione already.The 2-[1 that will make according to WO95/32176,1-dimethyl-2-acrylic]-3-hydroxyl-1, (500mg 2.06mmol) is dissolved in the concentrated sulfuric acid (10ml) to the 4-naphthoquinones, at room temperature stirs 10 minutes, then in the impouring frozen water (100ml).This aqueous mixtures with extracted by ether (3 * 50ml), and with the ether layer water that merges (2 * 50ml) and saturated nacl aqueous solution (50ml) wash, use dried over mgso then.Filter gained solution, steaming desolventizes, and product obtains orange-red needle-like dunnione (476mg), mp98-99 ℃ with benzinum (60-80 ℃) recrystallization.Embodiment 2-extracts dunnione

The process of extracting dunnione from slipperwort platymiscium (Calceolaria Integrifolia) can be carried out according to the method (Helvetica Chimica Acta, 60,945-947, (1977)) of R ü di and Evgster.The process of extracting dunnione from streptocarpus plant (Streptocarpusdunnii) can be carried out according to the method (J.Chem.Soc., (1939), 1522) of Price and Robinson.Embodiment 3 and 4-measure the antifungal activity of dunnione

The research dunnione is to being grown in the antifungal activity of 14 days fungies on the seedling age seedling under the conditions in vitro.The fungi strain of being studied among the embodiment 3 has wheat septoria (Septoriatritici), barley moire spore (Rhynchosporium secalis), Ustilago maydis (Ustilago maydis), coarse chain spore hail (Neurospora crassa), gray botrytis (Botrytis cinerea), F.graminearum schw (Fusarium graminearum), eye spot bacterium Pseudocercosporella herpotrichoides (eyespot), standing grain powdery mildew (Erysiphe graminis f.sp hordei).The dosage level of observation post's research compound also draws the conclusion of relevant their antifungal activities thus to the effect of above-mentioned fungi strain bacterium colony size.Embodiment 3-in vitro test

Fungus colony is gone up cultivation and growth at potato dextrose agar (PDA), and autoclaving is also used the compound treatment of studying, and potato dextrose agar wherein is by stock solution (20mg ml ^-1) at ethanol: preparation in the acetone (1: 1).For each compound, prepare four times of dilution series concentration, dosage range is from 20 μ g ml ^-1To 0.08 μ g ml ^-1With the fungi of yeast shape bacterium colony form growth, its growth is measured with minimum inhibitory concentration (MIC) for only; For other strain, measure extent of growth according to colony diameter, and calculate 50% effective dose (ED according to dose response curve ₅₀) value.In one of campaign, use Czapek-DOX to add fungi peptone (1%w/vCDM) and replace PDA.With each fungi incubation growth under optimum temperature.And relatively dunnione and the antifungal activity of the known antifungal compound of thing in contrast.Result shown in the following table 1 (a)-(g) shows that the antifungal activity of test compounds is relevant with the character of bacterial strain uses therefor.Dunnione is to wheat septoria (Septoria tritici), and it is active that barley moire spore (Rhynchosporium secalis) and Ustilago maydis (Ustilago maydis) have opposing; The activity level of anti-these pathogens of dunnione is similar to the azoles fungicide that uses at present or be better than them.Dunnione also demonstrates some activity to gray botrytis (Botrytis cinerea) and F.graminearum schw (Fusarinm graminearum); This two classes pathogen adopts present azoles fungicide to be difficult to control.Also observe significant anti-coarse chain spore hail (Neurospora crassa) activity in addition.Table 1 (a) wheat septoria (Septoria tritici)

	Sensitivity MIC (μ g/ml)
Separator			Flutriafol	Dunnione
S27	40	5
RL2	1	5

Table 1 (b) barley moire spore (Rhynchosporium secalis)

	Sensitivity MIC (μ g/m1)
					Separator	Carbendazim	Triadimenol	Propiconazole	Dunnione
CDM agar
					????81	????0.1	????＞50	????2.0	????5
????86	????0.1	????3.2	????0.4	????＞5
					????97	????0.1	????＞50	????＞10	????5
????101	????0.1	????＞50	????＞10	????＞5
					????110	????0.1	????＞50	????2.0	????＞5
????115	????0.1	????1.6	????0.08	????20
					????117	????0.1	????12.5	????2.0	????5
????118	????0.1	????＞50	????＞10	????20
					????120	????100	????＞50	????＞10	????20
PDA
					????810	????0.1	????＞50	????＞10	????20

Table 1 (c) Ustilago maydis (Ustilago maydis)

	ED 50(μg／ml)
Separator	Dunnione
The ATCC-14826 wild type	3.0

The coarse chain spore of table 1 (d) hail (Neurospora crassa)

	ED 50(μg／ml)
Separator	Dunnione
Am 132a	2

Table 1 (e) gray botrytis (Botrytis cinerea)

			Sensitivity ED 50(μg／ml)
				Separator	Dicarboximide	Benzimidazole	Dunnione
R3B6	R	R	＞20
RW10	S	S	＞20

S=susceptibility; R=resistance table 1 (f) F.graminearum schw (Fusarium graminearum)

		Sensitivity ED 50(μg／ml)
		Separator	Benzimidazole	Dunnione
JF18	R	20

R=resistance table 1 (g) eye spot bacterium (Pseudocercosporella herpotrichoides)

		Sensitivity ED 50(μg／ml)
		Separator	Benzimidazole	Dunnione
CR03A	R	17

The further in vitro test of R=resistance embodiment 4-

Adopt the test of method described in the embodiment 3 dunnione to black aspergillus (Aspergillusniger), the bactericidal activity of the separator of Rice Blast Fungus (Pyricularia oryzae) and Solanum rhizoctonia (Rhizoctonia solani).Handle for each, all will test on the agar of every kind of fungi in culture dish, quadruplicate, three parts of science bacterium colonies (then a for R.slani) are arranged in each flat board of agar.Black aspergillus (A.niger) and Solanum rhizoctonia (R.solani) were cultivated 4 days at 20-25 ℃; Rice Blast Fungus (P.oryzae) was then cultivated 7 days.Measure the bacterium colony activity according to the value added of colony diameter then.

Also to four kinds of separators of eye spot bacterium (Pseudocercosporella herpotrichoides), promptly two kinds of R-types of slowly growing and two kinds of W-type separators of growing are fast tested with dunnione.A kind of separator in every type is to the bactericide sensitivity such as carbendazim and prochloraz, and another kind then is a complete resistance.At two week of 20 ℃ of growths back measurement colony diameter.

Also four kinds of separators of potato tubers pathogen Helminthosporium solani (Helminthosporium solani) and sulphur look sickle spore (Fusarium sulphureum) are tested with dunnione.Two kinds of separators in every kind of pathogen are to bactericide thiabendazole (TBZ) sensitivity, other two kinds of resistances that then have to a certain degree.Then carry out and the used same steps as of eye spot bacterium (P.Herpotrichoides), but just replica is grown on malt extract agar, and measure growing state twice.

Table 2 (a) shows that to the result shown in 2 (f) dunnione is to black aspergillus (Aspergillus niger), and Rice Blast Fungus (Pyricularia oryzae) and Solanum rhizoctonia (Rhizoctonia solani) have activity.Its anti-eye spot bacterium (Pseudocercosporella herpotrichoides) activity is volatile.Do not have evidence to show that dunnione and carbendazim (table 2 (d)) have cross resistance, or Helminthosporium solani (H.solani) or sulphur look sickle spore (F.sulphureum) are had cross resistance (table 2 (e) and (f)) with thiabendazole.The anti-black aspergillus of table 2 (a)-dunnione (Aspergillus niger) activity

Compound	The inhibition % of colony diameter growth
					100 mg／l	20 mg／l	4 mg／l	1 mg／l
Dunnione	100	99	30
Prochloraz				97.1

Table 2 (b)-dunnione antipiriculin (Pyricularia oryzae) activity

Compound	The inhibition % of colony diameter growth
					100 mg／l	20 mg／l	4 mg／l	1 mg／l
Dunnione	100	99	75
Carbendazim			99.4	14.7

The anti-Solanum rhizoctonia of table 2 (c)-dunnione (Rhizoctonia solani) activity

Compound	The inhibition % of colony diameter growth
					100 mg／l	20 mg／l	4 mg／l	1 mg／l
Dunnione	100	93	37
Carbendazim			81.2	3.3

The activity of W-type of the anti-eye spot bacterium of table 2 (d)-dunnione (Pseudocercosporellaherpotrichides) and R-type separator

The fungi type	Susceptibility to carbendazim	Activity under the 5mg/ml consumption
				Carbendazim	Dunnione
The W-type	Responsive	100	50
The W-type	Resistance	5	30
The R-type	Responsive	100	-15
The R-type	Resistance	-14	4

Table 2 (e)-dunnione is to having the activity of the Helminthosporium solani of resistance (Helminthosporium solani) separator in various degree to thiabendazole (TBZ)

Separator	Activity under the 5mg/ml consumption
						8 days		14 days
	Thiabendazole	Dunnione	Thiabendazole	Dunnione
					????S1	????100	????65	????100	????53
????S2	????100	????63	????100	????86
					????R1	????43	????57	????20	????87
????R2	????100	????70	????100	????56

S1, S2=is to the separator of thiabendazole sensitivity; R1, R2=have the separator of resistance to thiabendazole.

Table 2 (f)-dunnione is to having the activity of sulphur look sickle spore (Fusarium sulphureum) separator of resistance in various degree to thiabendazole

Separator	Activity under the 5mg/ml consumption
						8 days		14 days
	Thiabendazole	Dunnione	Thiabendazole	Dunnione
					????S1	????100	????22	????100	????nd
????S2	????100	????11	????100	????5
					????R1	????60	????4	????64	????6
????R2	????70	????16	????70	????12

S1, S2=is to the separator of thiabendazole sensitivity; R1, R2=have the separator of resistance to thiabendazole.The nd=free of data; Fungus colony in the control group medium reaches the edge of culture plate.Embodiment 5-14 days seedling age seedling

Before conidium inoculation, 14 days seedling age seedling were sprayed 24 hours with standing grain powdery mildew (Erysiphe graminis f.sp hordei).The dunnione solution spray that will make in rare polysorbas20 solution (1 polysorbas20/100ml water) is to seedling; The ultimate density of dunnione is 0.75mg ml in the used solution ^-1With 0.075mg ml ^-1, preceding a kind of concentration is equivalent to the land for growing field crops amount of application of triazole bactericide.The control group seedling is only sprayed polysorbas20 solution.Inoculate the growing state of the percentage assessment standing grain powdery mildew that infects according to leaf area after 7 days.Given presentation of results in the table 3, dunnione still has effective bactericidal activity under the dosage level that is lower than at present used triazole bactericide, and does not produce tangible poisoning.When handling some kind plant, because higher concentration may produce some poisoning, thereby preferred the use is lower than 0.75mg ml ^-1Concentration.Table 3-dunnione is to the influence of standing grain powdery mildew (Erysiphe graminis f.sp hordei) growth

		Disease control %
				Separator	Triadimenol	Dunnione (mg/ml)
		0.75	0.075
23D5	S	100	45
212	R	100	50

The S=sensitivity; The R=resistance.

Claims (16)

1. formula (I) dunnione:

Application as bactericide.

2. the enantiomer of dunnione or mixture of enantiomers are as the application of bactericide.

3. bactericidal composition, said composition comprises inert carrier and dunnione.

4. according to the bactericidal composition of claim 3, inert carrier wherein is selected from water, aliphatic series or aromatic hydrocarbon and alcohol.

5. according to the bactericidal composition of claim 4, aliphatic series wherein or aromatic hydrocarbon are dimethylbenzene and other petroleum distillate.

6. according to the bactericidal composition of claim 4, wherein said alcohol is ethanol.

7. according to each bactericidal composition among the claim 3-6, said composition also further comprises one or more surfactants.

8. according to each bactericidal composition among the claim 3-7, said composition further comprises stabilizing agent.

9. according to each bactericidal composition among the claim 3-8, said composition further comprises one or more other fungicide active ingredients.

10. the method for preparing bactericidal composition, this method comprise the step of mixing dunnione and appropriate carrier.

11. the method that control or prevention fungi are invaded and harassed, this method comprise the step that the bactericidal composition that makes claim 9 contacts with fungi.

12. the material of handling with each described dunnione among the claim 1-9.

13. according to the material of claim 12, the material of wherein said processing is a vegetable material.

14. plant extract as the application of bactericide, is characterized in that comprising in the plant extract formula (I) dunnione:

15. according to the application of claim 14, wherein plant extract derives from streptocarpus plant Streptocarpus dunnii.

16. according to the application of claim 14, wherein plant extract derives from slipperwort platymiscium Calceolaria Integrifolia.