CN102795969A - Natural sesquiterpene guaiol compound and medicinal application thereof - Google Patents

Abstract

The present invention belongs to the field of pharmacy and relates to natural sesquiterpene compound guaiol, in particular to the natural sesquiterpene compound guaiol (guaiol) of formula (I), its preparation method and its use in the preparation of plant pesticides . The results of the insecticidal experiment of the compound of the present invention show that the compound guaiacol of the present invention exhibits antifeedant, contact killing, fumigation and juvenile hormone analogue activities to the test insects, and its mode of action is closely related to the dosage; The contact toxicity LD ₅₀ of Plutella xylostella and 3rd instar Plutella xylostella were 0.072mg/head, 24h and 8.920mg/head, 24h respectively; the fumigation toxicity LC ₅₀ to Musca domestica and 4th instar armyworm adults were 3.52μL/L, respectively, 12h and 16.95μL/L, 12h, the results show that the compound has better insecticidal activity and can be used as an insecticidal active component to further prepare plant insecticides such as sprays or fumigants. (I)

Description

Natural sesquiterpene compound guaiacol and its medicinal use

technical field

The invention belongs to the field of pharmacy and relates to a natural sesquiterpene compound guaiol, in particular to a natural sesquiterpene compound guaiol (guaiol), a preparation method thereof and an application in preparing plant pesticides.

Background technique

With the emergence of problems such as environmental pollution, pesticide residues and pest resistance, as well as the rapid development of organic agriculture in the world, higher requirements have been placed on the pesticide industry. It is known that the direction of pesticide development is to develop environment-friendly pesticides with high efficiency, low toxicity, low residue and low resistance risk, and botanical pesticides meet this development requirement. Generally speaking, botanical pesticides come from nature and have a smooth degradation path in the environment, so they do not pollute the environment and have little adverse impact on the environment; practice shows that their biological activities are diverse, not only have insecticidal activity, but also often It has the functions of sterilizing and regulating plant growth, and has various forms of action and unique mechanism of action. It can have multiple targets in the pest body, so pests are not easy to develop drug resistance, and there is no cross-resistance with existing conventional chemical insecticides. : Strong selectivity to pests, safe to natural enemies and wild life. In the past 30 years, relevant researchers have discovered many substances with specific biological activities from plants, and have achieved great success in insecticidal spectrum and toxicity mechanism. Foreign studies have evaluated the insecticidal effects of Zingiberaceae plants, Araceae plants, Lauraceae plants and other plants, and achieved good results. At present, it is recognized that the pyrethrins and carbamates in the three mature insecticides and the neonicotinoid insecticides developed rapidly in recent years all have their synthetic lead compounds derived from plants. Therefore, research on botanical insecticides has become a focus.

The mechanism of action of plant pesticides includes affecting the growth and development of insects and the digestive system; affecting the respiratory system of insects; affecting the nervous system of insects and affecting the neuromuscular system of insects, etc. Insecticidal active ingredients in plants are secondary metabolites formed during the long-term co-evolution of plants and insects. They have unique symptoms, diverse modes of action, and complex mechanisms of action on pests. Generally speaking, such substances will not pollute the environment, are relatively safe for humans, livestock, crops, and the ecological environment, and insects are not easy to develop resistance to them. These characteristics meet people's requirements for ideal insecticides, and also meet the needs of sustainable agriculture. development strategy requirements. Researchers believe that finding new insecticidal active substances from plant secondary compounds is one of the effective ways to develop new insecticides and overcome or delay the emergence of drug resistance.

The prior art relevant to the present invention has:

[1] Cong Dapeng. Research and problem analysis of botanical pesticides in my country [J]. Pesticide Research and Application, 2009, 13 (1): 7-9.

[2] Pandji C. Insecticidal constituents from species of the Zingiberacene[J]. Phytochemistry, 1993, 34 (2): 415-419.

[3] Suzuki K. Eight picrotoxane terpenoids, picrodendrius K-R, from pcrodendron banatcrm[J]. Phytochemistry, 1992, 31(6): 2059-2064.

[4] Gonzalez-coloma A, Cabera R, Monzon A R S. Persea indica as a natural source of the insecticide ryanodol[J]. Phytochemistry, 1993, 34 (2): 397-400.

[5] Fu Xiuqin. Survey of Plant Pesticides Research [J]. Journal of Wuhan Institute of Bioengineering, 2006, 2(1): 55-59. .

Contents of the invention

The purpose of the present invention is to provide a kind of new botanical insecticide, be specifically related to natural sesquiterpene compound guaiol (guaiol), and medicinal use thereof. The compound of the present invention is ferulic natural sesquiterpene compound guaiacol, which has insecticidal activity and can be further made into plant insecticide.

The compound of the present invention is a guaiac-type sesquiterpene, which has a structure shown in formula (I), and its structural feature is that the compound has a seven-membered ring and a five-membered ring skeleton, the 1-position of the ring is a double bond, 3a , 8a, 5-position is substituted by hydrocarbon group, other positions are unsubstituted, its molecular weight is 222, its molecular formula is C ₁₅ H ₂₆ O; its chemical structure is:

2-[(3S*,5R*,8S*)-3,8-dimethyl-1,2,3,4,5,6,7,8-octahydroazulen-5-yl]propan-2-ol.

the

                                                         

(I)

Guaiacol, the compound of this invention, is obtained from the medicinal plant Ferula spp. in Xinjiang through sub-extraction and separation: take the root part of the dried plant Ferula spp., crush it, and use 20 times the amount of 95% ethanol at 45 ^o C Soaking and extracting, the extracts were combined and evaporated to dryness under reduced pressure to obtain the extract, the obtained extract was extracted with dichloromethane, the organic phase was evaporated to dryness under reduced pressure to obtain the extract, and the obtained extract was recrystallized after several times of normal phase silica gel column chromatography The compound guaiol is obtained.

The structure of the obtained compound is determined by the determination of physical constants and spectral data analysis, and is proved by X-ray single crystal diffraction. The determination method is: the melting point is measured by WRX-4 melting point apparatus; the specific rotation value [α] is measured by Polarimeter P-1020, JASCO Corporation type polarimeter; the mass spectrum is measured by Agilent 5973N MSD type mass spectrometer; the carbon and hydrogen spectrum data are determined by arian Measured by Mercury Plus 400MHz superconducting nuclear magnetic resonance instrument, the solvent is deuterated chloroform; the X-ray single crystal diffraction conditions are as follows: CAD4 DIFFACTIS 586 single crystal diffractometer, the selected crystal size is 0.12 x 0.10 x 0.08 mm, and the ray is Mo/Kα (λ=0.71 073 A), scanned in ω-θ mode, the scanning range is 1.80°<θ<27.84°, and the diffraction points were collected at 293(2)K; the intensity data were corrected by LP and empirical absorption, and the direct method and The Fourier synthesis method was used to solve the crystal structure, and the coordinates and anisotropy parameters of all non-hydrogen atoms were corrected by the full matrix least squares method; the crystal structure analysis was completed on a PC computer with the program Shelxl-97.

Physical property and spectral data of compound guaiac alcohol (guaiol) of the present invention are as follows:

guaiol: white blocky crystals, easily soluble in acetone, chloroform, etc.; molecular formula C ₁₅ H ₂₆ O(222); EIMS m/z 222[M] ⁺ ; [α] ¹⁶ _D : -52.0 (c 0.500, CHCl ₃ ) ;mp 90-91 ^o C. ¹ H NMR (CDCl ₃ ) δ: 2.11, 2.43 (2H, m, 1-H), 1.28, 1.97 (2H, m, 2-H), 2.54 (1H, m, 3-H), 1.83, 2.13 ( 2H, m, 4-H), 1.56 (1H, m, 5-H), 1.58, 1.72 (2H, m, 6-H), 1.46, 1.76 (2H, m, 7-H), 2.29 (1H, m, 8-H), 1.17 (3H, s, 1'-H), 1.19 (3H, s, 3'-H), 0.95 (3H, s, 3-CH ₃ ), 0.98 (3H, s, 8 -CH ₃ ); ¹³ C NMR (CDCl ₃ ) δ: 35.4 (C-1), 30.9 (C-2), 46.3 (C-3), 138.9 (C-3a), 27.8 (C-4), 49.6 (C-)5, 33.7 (C-6), 27.3 (C-7), 33.7 (C-8), 140.0 (C-8a), 26.0 (C-1'), 73.5 (C-2'), 27.4 (C-3'), 19.9 (3-CH ₃ ), 19.7 (8-CH ₃ ).

The structure of the compound obtained by analyzing the spectral data is: 2-[(3S*,5R*,8S*)-3,8-dimethyl-1,2,3,4,5,6,7,8-octahydroazulen-5-yl] propan-2-ol.

In the present invention, the medicinal plant can be Ferula ferulaeoides (Steud.) Korov., Ferula sinkiangensis or Ferula fukanensis, which are plants of the genus Ferula in the family Umbelliferae.

The invention provides the use of the natural sesquiterpene compound guaiol in the preparation of plant pesticides.

The present invention has been proved by experiments that the ferulic extract guaiacol has antifeedant, contact killing, fumigation and juvenile hormone analogue activities on the test insects, and its mode of action is closely related to the dosage; Armyworms hardly feed and show symptoms of poisoning such as excitement, convulsions, paralysis, and death successively. After gradually reducing the concentration, the food intake of armyworms increases gradually; in addition, the results also show that ferulic extract has a positive effect on Tenebrio molitor pupae. Similar to the juvenile hormone, and the effect on the growth and development of insects.

The present invention measures the insecticidal activity of guaiacol by contact killing and fumigation insecticidal experimental method:

(1) Drop method for contact activity: dilute the test agent with acetone to the required concentration, and quantitatively drop the drug on the pronotum of the test larvae with a microcapillary tube, and drop 10 larvae for each treatment. The amount of acetone was used as a control. After spot treatment, the test insects were reared in the insect culture room [T=(25±1)°C, RH=75%±5%, D/L=12h/12h], and the number of dead insects was investigated after 24 hours. Calculate the adjusted mortality rate.

The selected experimental pests are: armyworm (Mythimna separata Walker) and diamondback moth (Plutella xylostella Linnaeus); (2) fumigation activity with triangular flask fumigation method: fumigation container with 250ml triangular flask (volume is 330m1), fixed with a pin

Seal the rubber stopper of 9 cm × l cm filter paper, place 10 test insects in the triangular flask and give a small amount of food, dilute the test agent with acetone to the required concentration, and measure 1 μL-5 μL with a micro-sampler Add the liquid medicine dropwise on the filter paper, quickly cover the triangular flask tightly, investigate the number of dead insects 12 hours after the treatment, 10 test insects are treated as one, and each treatment is repeated 3 times, and the mortality rate and corrected mortality rate are calculated;

The selected experimental pests are: armyworm (Mythimna separata Walker) and housefly (Eusca domestica Linnaeus);

Calculate the toxicity curve and lethal median dose of the compound to the test insects by the probability value analysis method;

Insecticidal experiment result shows: compound guaiac of the present invention (guaiol) is respectively 0.072 mg/ head (24 h) and 8.920 mg/ head (24 h) to the contact toxicity LD ₅₀ of 4 instar armyworm and 3 instar diamondback moth. h); the fumigation toxicity LC ₅₀ to housefly and 4th instar armyworm adults is respectively 3.52 μL/L (12 h) and 16.95 μL/L (12 h), the results show that the insecticidal activity of the compound is higher than Well, it can be used as an insecticidal active component to further prepare plant pesticides. The plant insecticide is made into types such as spray or fumigant.

Description of drawings

Figure 1 is a single crystal X-ray diffraction pattern of the compound of formula (I).

Detailed ways

Embodiment 1 prepares guaiol (guaiol) by ferulicum ferulae

Dried plant Ferula ferulaeoides (Steud.) Korov. Root parts 2.0 kg, crushed, soaked and extracted with 20 times the amount of 95% ethanol at 45 ^o C for 3 times, combined the extracts and evaporated to dryness under reduced pressure cream, the obtained extract was extracted three times with dichloromethane, and evaporated to dryness under reduced pressure to obtain 400 g of extract. The sample was subjected to silica gel column chromatography, using 200-300 mesh silica gel (Qingdao Ocean Chemical Factory, China), petroleum ether-ethyl acetate (100:0 ~ 0:100) gradient elution, and silica gel thin-layer chromatography detection. Combine the fractions eluted with petroleum ether-ethyl acetate 97:3, evaporate to dryness to obtain a white solid, and repeatedly use 300-400 mesh silica gel column chromatography, the eluent is petroleum ether-acetone (99:1 ~ 97:3), and finally Recrystallization gave about 1.6 g of guaiol in the form of white blocky crystals. It was detected as a pure compound by forward and reverse phase silica gel thin-layer chromatography and HPLC, and was identified as the compound guaiol (guaiol) of formula (I) by spectral data.

Example 2 Contact test of compound (I) on armyworm and diamondback moth

Dilute the reagent to be tested with acetone to the required concentration, and use a micro capillary to quantitatively drip the drug on the pronotum of the larvae of the test insects, and drip 10 larvae for each treatment. Insects are reared in the insect culture room [T=(25±1)°C, RH=75%±5%, D/L=12h/12h], and the number of dead insects is investigated after 24 hours, and the corrected mortality rate is calculated (as shown in Table 1. Show).

Table 1 is the contact toxicity of guaiol (guaiol) to armyworm and diamondback moth.

the

Table 1

Example 3 Compound (I) fumigation insecticidal experiment on armyworm and housefly

The fumigation container is with a 250ml triangular flask (volume is 330m), and a rubber stopper of 9 cm × 1 cm filter paper is fixed with a pin and sealed; 10 test insects are placed in the triangular flask and a small amount of food is given, and the test agent is diluted to the desired level with acetone. After the concentration is required, use a micro-sampler to measure 1 μL-5 μL of the drug solution and drop it on the filter paper, quickly cover the triangular flask tightly, and investigate the number of dead insects 12 hours after the treatment; 10 test insects are treated as one, and each treatment is repeated Three times to calculate the mortality rate and corrected mortality rate; use the probability value analysis method to obtain the toxicity curve and the lethal median dose of the essential oil to the test insects.

Activity test results showed that the contact toxicity LD ₅₀ of the compound guaiol to 4th instar armyworm and 3rd instar diamondback moth was 0.072 mg/head (24 h) and 8.920 mg/head (24 h); The fumigation virulence LC ₅₀ of instar armyworm adults were 3.52 μL/L (12 h) and 16.95 μL/L (12 h) respectively (as shown in Table 2).

Table 2 is the fumigation toxicity of guaiol to armyworm and housefly.

Table 2

Claims (5)

1. The compound guaiacol of formula (I) structure is characterized in that, this compound has seven-membered ring and five-membered ring skeleton, ring 1-position is a double bond, 3a, 8a, 5-position are substituted by hydrocarbon groups, No substitutions in other positions,                                                         

(I). 2. by the described compound guaiacol of claim 1, it is characterized in that, its molecular weight of described compound is 222, and molecular formula is C ₁₅ H ₂₆ O; Its chemical structure is: 2-[(3S*,5R*,8S*)-3,8-dimethyl-1,2,3,4,5,6,7,8-octahydroazulen-5-yl]propan-2-ol. 3. The preparation method of the compound guaiacol described in claim 1 is characterized in that, it is obtained by sub-extraction and separation of the medicinal plant Ferula spp. in Xinjiang: take the root part of the dried plant Ferula spp., pulverize it, and use 20 Double the amount of 95% ethanol was soaked and extracted at 45 ^o C, the extracts were combined and evaporated to dryness under reduced pressure to obtain the extract, the obtained extract was extracted with dichloromethane, and the organic phase was evaporated to dryness under reduced pressure to obtain the extract. The compound guaiol was obtained by recrystallization after normal phase silica gel column chromatography. 4. A natural plant insecticide, characterized in that the compound guaiacol according to claim 1 is used as the insecticidal active component. 5. according to the natural plant insecticide of claim 4, it is characterized in that, described natural plant insecticide is used for killing insects: armyworm Mythimna separata Walke, diamondback moth Plutella xylostella Linnaeu) and housefly Eusca domestica Linnaeus.

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