Novel glycosyl C-glycoside thiadiazoline derivative as well as preparation method and application thereof

Abstract

The invention provides a novel glycosyl C-glycoside thiadiazoline derivative as well as a preparation method and application thereof, and belongs to the field of pesticides. The structural formula is shown as a formula (I) or a formula (II). According to the invention, the glycosyl carbon glycoside compound with a novel structure is synthesized by taking the insect chitin degrading enzyme as a biological target. The glycosyl C-glycoside compound or the pharmaceutically acceptable salt thereof has good target enzyme inhibition activity and insecticidal activity, and the insecticidal activity of part of the compound to plutella xylostella and ostrinia nubilalis is equivalent to that of a control agent diflubenzuron. The compound disclosed by the invention has a very high pesticide research value. # imgabs0 #

Description

Technical Field

[0001] The invention belongs to the field of pesticides, and specifically relates to a novel glycosyl carbonyl thiadiazoline derivative and a preparation method and application thereof. Background Art

[0002] Lepidoptera agricultural pests cause serious damage to crops such as corn, sorghum, and millet, seriously affecting the quality and yield of crops and causing huge economic losses every year. Chitin is a linear polysaccharide connected by β-1,4-glycosidic bonds. It is the main structural component of the cuticle of lepidopteran insects and plays an important role in protecting insects from various stimuli and damage from the external environment. Lepidopteran pests will periodically molt through the degradation of chitin throughout their life cycle, which is necessary for the normal growth and development of lepidopteran pests. Inhibition of enzymes that degrade and metabolize the cuticle can cause abnormal molting of lepidopteran insects, thereby inhibiting the growth and development of insects and even leading to death. In the process of chitin degradation, chitinase (EC 3.2.1.14) of glycohydrolase family 18 (GH18) and β-N-acetylglucosaminidase (Hex; EC3.2.1.52) of glycohydrolase family 20 (GH20) play an important role in chitin degradation. Specifically, chitinase can effectively convert long-chain chitin into short-chain chito-oligosaccharides. β-N-acetylhexosaminidase further hydrolyzes chito-oligosaccharides into monosaccharides, thus achieving the chitin degradation process. Therefore, designing new pesticides targeting GH18 chitinase and GH20 β-N-acetylhexosaminidase is a potential pest control strategy. Summary of the invention

[0003] The present invention provides a novel glycoside compound containing N-acetylglucosamine, thiosemicarbazide or 1,3,4-thiadiazole structure or a pharmaceutically acceptable salt, solvate or hydrate thereof and a preparation method thereof.

[0004] The glycoside compounds provided by the present invention have a structural formula as shown in formula (I) or formula (II), which are respectively denoted as CAUZA-A and CAUZA-B:

[0005]

[0006] In formula (I) and formula (II), R is substituted or unsubstituted phenyl, benzyl or naphthyl;

[0007] The substitution may be mono- or poly-substituted.

[0008] The position of the mono-substitution may be the 2-position, the 3-position or the 4-position; the poly-substitution may be di-substitution, and the position of the di-substitution may be the 3-position and the 4-position;

[0009] The substituents are each independently selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, C1-C6 alkyl-substituted amino;

[0010] Further, each of the substituents is independently selected from halogen, C1-C3 alkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, C1-C3 alkyl-substituted amino;

[0011] Furthermore, each of the substituents is independently selected from any one of chlorine, bromine, methyl, trifluoromethyl, methoxy, N,N-dimethylamino and N,N-diethylamino.

[0012] Specifically, the glycoside compound represented by formula (I) is any one of the following compounds:

[0013]

[0014] The glycoside compound represented by formula (II) is any one of the following compounds:

[0015]

[0016] The compound represented by the above formula (I) is prepared by a method comprising the following steps:

[0017] (1) reacting the compound represented by formula (III) with the compound represented by formula (IV) in an ethanol solvent to obtain a compound represented by formula (V);

[0018]

[0019] The definition of R in formula (III) is the same as that of R in formula (I);

[0020] The definition of R in formula (V) is the same as that of R in formula (I);

[0021] (2) Deprotecting the compound represented by formula (V) under methanol-ammonia conditions to obtain the target compound represented by formula (I).

[0022] The compound represented by the above formula (II) is prepared by a method comprising the following steps:

[0023] 1) reacting the compound represented by formula (V) with active manganese dioxide in chloroform to obtain the compound represented by formula (VI);

[0024]

[0025] The definition of R in formula (VI) is the same as that of R in formula (II);

[0026] 2) Deprotecting the compound represented by formula (VI) under methanol-ammonia conditions to obtain the target compound represented by formula (II).

[0027] The compound represented by formula (V) and the compound represented by formula (VI) in the above method also fall within the protection scope of the present invention.

[0028] The present invention also provides the use of the glycosyl carbonyl glycoside thiosemicarbazide represented by formula (I) and the glycosyl carbonyl glycoside thiadiazoline derivative represented by formula (II) or their pharmaceutically acceptable salts, solvates, hydrates or compositions containing the same as insect chitin degrading enzyme inhibitors in the preparation of insecticides.

[0029] In the application, the insects are agricultural pests of the order Lepidoptera, including Plutella xylostella and Ostrinia nubilalis.

[0030] The present invention also provides an insecticide, which contains the glycosyl carbonyl glycoside semicarbazide represented by formula (I) or the glycosyl carbonyl glycoside thiadiazoline derivative represented by formula (II) or a pharmaceutically acceptable salt, solvate, hydrate or a composition containing the same.

[0031] The insecticide is in a pharmaceutically acceptable dosage form, such as emulsifiable concentrate, wettable powder, suspension, dust, soluble powder, aqueous solution, water-dispersible granules, smoke agent, granules, seed coating agent, etc.

[0032] For example, an insecticide emulsifiable concentrate is composed of the following substances in percentage by mass: 1-10% of the above-mentioned glycoside compound or a pharmaceutically acceptable salt thereof, 5-15% of an emulsifier, 0.1-1% of a penetrant, and the remainder being a solvent.

[0033] The emulsifier is a surfactant, such as Nongru 0203B, 0208, GFC, OP-10, Tween-60, etc.

[0034] The solvent may be toluene, xylene, or the like.

[0035] For example, a pesticide wettable powder is composed of the following substances in percentage by mass: 15-50% of the above-mentioned glycoside compound or a pharmaceutically acceptable salt thereof, 10-20% of a surfactant (such as surfactant NNO) and 30-75% of white carbon black.

[0036] The inventors of the present invention used GH18 chitinase and GH20 β-N-acetylhexosaminase of Asian corn borer (Ostriniafurnacalis) as targets, designed glycosylnaphthylimide glucosinolate as lead compounds, replaced the glucosidic bond with a more stable carbon glycosidic bond, connected acetylglucosamine with different aromatic groups through thiosemicarbazide or 1,3,4-thiadiazole active fragments, and synthesized a series of compounds with good activity and selectivity to achieve good enzyme inhibition activity. Finally, through insecticidal activity tests, it was found that this series of compounds had good insecticidal activity against Asian corn borer and diamondback moth.

[0037] The beneficial effects of the present invention are as follows: the present invention uses insect chitin degrading enzyme as a biological target to synthesize a novel structural glycoside compound. The glycoside compound or a pharmaceutically acceptable salt thereof of the present invention has good target enzyme inhibitory activity and insecticidal activity, and the insecticidal activity of some compounds against diamondback moth and corn borer is equivalent to that of the control agent diflubenzuron. The compound of the present invention has a high pesticide research value. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 This is the hydrogen nuclear magnetic resonance spectrum of the compound CAUZA-A-01 prepared in the present invention.

[0039] Figure 2 This is the carbon nuclear magnetic resonance spectrum of the compound CAUZA-A-01 prepared in the present invention.

[0040] Figure 3 This is the hydrogen nuclear magnetic resonance spectrum of the compound CAUZA-B-01 prepared in the present invention.

[0041] Figure 4 This is the carbon nuclear magnetic resonance spectrum of the compound CAUZA-B-01 prepared in the present invention. DETAILED DESCRIPTION

[0042] The present invention is further described in detail below in conjunction with specific embodiments, and the examples provided are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can be used as a guide for further improvements by those of ordinary skill in the art, and do not constitute a limitation of the present invention in any way.

[0043] The experimental methods in the following examples, unless otherwise specified, are all conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples, unless otherwise specified, can all be obtained from commercial channels.

[0044] Example 1. Preparation and structural identification of compound CAUZL-A-01, R=Ph.

[0045]

[0046] In a 50 mL round-bottom flask, add 1-C-(2-acetylamino-2-deoxy-3,4,6-tri-O-acetyl-β-D-pyranose)-propan-2-one (1.0 g, 2.58 mmol); 4-phenyl-3-thiosemicarbazide (0.43 g, 2.58 mmol, 1.0 equivalent) and 20 ml of anhydrous ethanol. Stir at 80°C for 8 hours, concentrate after the reaction is completed, and the concentrate is purified by column chromatography to obtain 0.94 g of a white solid with a yield of 67.9%.

[0047] Structure confirmation data: 1 H NMR(500MHz,Chloroform-d)δ9.47(s,1H,-NH-),8.60(s,1H,-NH-),7.63(d,J= 7.6Hz,2H,-PhH),7.33–7.26(m,2H,-PhH),7.18–7.11(m,1H,-PhH),6.00(d,J= 9.3Hz,1H,-NH-),5.07–4.92(m,2H,-CH2-),4.18–3.89(m,3H,H-1,H-3,H-4),3 .68–3.49(m,2H,H-2,H-5),2.64–2.42(m,2H,-CH2-),2.02–1.81(m,15H,-CH3). 13 C NMR (125MHz, CDCl3) δ205.37,175.37,175.31,170.61,170.56,170.48,169.87,169.84,169.73,169.65,169.48 ,169.22,168.36,168.31,168.20,149.18,148.09,137.22,137.00,127.69,127.61,124.90,124.83,123.36,122 .94,76.55,74.88,74.81,74.10,73.26,73.22,72.74,67.38,67.33,67.07,61.35,61.22,61.10,59.41,53.12,52.62,52.11,44.95,39.17,32.93,30.06,28.67,23.71,22.31,22.15,19.88,19.71,19.61,19.59,16.09,13.18.

[0048] The product obtained in (1) (0.94 g, 1.7 mmol), 10 mL of methanol, and 10 mL of saturated methanolic ammonia solution were added to a 50 mL round-bottom flask. The mixture was reacted for 4 h, concentrated, and purified by column chromatography to obtain 0.69 g of a white solid with a yield of 95.6%.

[0049] Structure confirmation data: 1 H NMR(500MHz,Chloroform-d)δ10.00(d,J=442.1Hz,1H,-NH),9.21(s,1H,-NH),7.76–7.41(m,3H,-P hH),7.32(q,J=9.4,8.6Hz,2H,-PhH),7.18(q,J=11.4,9.5Hz,1H,-NH),5.62–5.06(m,2H,-OH),4.5 0(d,J=199.0Hz,1H,-OH),3.87(d,J=52.5Hz,2H,-CH2-),3.68(s,2H,H-1,H-3),3.56(d,J=35.5Hz, 2H,H-2,H-4),3.32(d,J=26.4Hz,1H,H-5),2.80–2.27(m,2H,-CH2-),2.01(d,J=10.9Hz,6H,-CH3). 13 C NMR (125MHz, CDCl3) δ175.76,172.49,172.21,153.02,138.42,137.67,128.75,128.61,126.13,124.36,80.05,7 7.33,75.37,70.46,69.65,60.30,55.39,35.24,30.85,29.71,24.92,23.52,23.43,17.17.HR-MS(ESI)m / z:calcd for C 18 H 26 N4O5S[M+H] + ,411.1702;found,411.1698.

[0050] Other series of compounds with the general formula CAUZL-A were prepared according to the above method. Their compound numbers, corresponding substituent groups, and physical and chemical data are shown in Table 1, and the nuclear magnetic resonance hydrogen spectrum and mass spectrum data for structural identification are shown in Table 2.

[0051] Table 1 Numbers, substitution groups, and physicochemical data of some compounds in the CAUZL-A series

[0052]

[0053] Table 2 H NMR and mass spectrometry data of some compounds in the CAUZA-A series

[0054]

[0055]

[0056]

[0057]

[0058] Example 2. Preparation and structural identification of compound CAUZA-B-01, R=Ph

[0059]

[0060] (1) Add (2R,3S,4R,5S,6S)-5-acetylamino-2-(acetoxymethyl)-6-((E)-2-(2(phenylaminomethoxy)hydrazino)propyl)tetrahydro-2H-pyran-3,4-diacetic acid diester (1.0 g, 1.86 mmol); active manganese dioxide (1.6 g, 18.6 mmol, 10.0 equivalents) and 20 ml of chloroform into a 50 mL round-bottom flask. Stir at room temperature for 5 hours. After the reaction is completed, filter to remove manganese dioxide, desolventize the filtrate under reduced pressure, and obtain 0.69 g of a yellow solid by column chromatography of the crude product, with a yield of 69.6%.

[0061] Structure confirmation data: 1 H NMR(500MHz,Chloroform-d)δ7.46(dd,J=8.8,7.0Hz,2H,-PhH),7.31–7.27(m,2H,-PhH),7.26(d,J=1.9Hz,1H,-PhH),5 .72(dd,J=9.3,3.0Hz,1H,-NH-),5.05–4.91(m,2H,-CH2-),4.15(dd,J=12.3,6.2Hz,1H,H-3),4.08–3.93(m,2H,H-1,H- 5),3.56(td,J=10.1,2.0Hz,1H,H-4),3.47(ddd,J=9.9,6.2,2.2Hz,1H,H-2),2.63(dd,J=15.5,10.1Hz,1H,-CH2-),2.3 6(dd,J=15.5,2.0Hz,1H,-CH2-),2.10(s,3H,-CH3),2.02(d,J=5.2Hz,6H,-CH3),1.94(s,3H,-CH3),1.83(s,3H,-CH3). 13C NMR(126MHz, CDCl3)δ174.52(s),171.60(s),170.88(s),170.44(s),169.29(s),148.09(s),129.48(d),126.97(s),120.95(d),108.3 8(s),76.16(s),75.95(s),73.96(s),68.37(s),62.35(s),53.68(s),41.56(s),27.28(s),23.25(s),20.81(s),20.65(s),20.59(s).

[0062] (2) The product obtained in (1) (0.69 g, 1.30 mmol), 10 mL of methanol, and 10 mL of saturated methanolic ammonia solution were added to a 50 mL round-bottom flask. The mixture was reacted for 4 h, concentrated, and purified by column chromatography to obtain 0.48 g of a yellow solid with a yield of 90.7%.

[0063] Structure confirmation data: 1 H NMR(500MHz,Chloroform-d)δ10.00(d,J=442.1Hz,1H,-NH),9.21(s,1H,-NH),7.76–7.41(m,3H,-P hH),7.32(q,J=9.4,8.6Hz,2H,-PhH),7.18(q,J=11.4,9.5Hz,1H,-NH),5.62–5.06(m,2H,-OH),4.5 0(d,J=199.0Hz,1H,-OH),3.87(d,J=52.5Hz,2H,-CH2-),3.68(s,2H,H-1,H-3),3.56(d,J=35.5Hz, 2H,H-2,H-4),3.32(d,J=26.4Hz,1H,H-5),2.80–2.27(m,2H,-CH2-),2.01(d,J=10.9Hz,6H,-CH3). 13 C NMR (125MHz, CDCl3) δ175.76,172.49,172.21,153.02,138.42,137.67,128.75,128.61,126.13,124.3 6,80.05,77.33,75.37,70.46,69.65,60.30,55.39,35.24,30.85,29.71,24.92,23.52,23.43,17.17.

[0064] Other series of compounds with the general formula CAUZA-B were prepared according to the above method. Their compound numbers, corresponding substituent groups, and physical and chemical data are shown in Table 3, and the nuclear magnetic resonance hydrogen spectrum and mass spectrum data for structural identification are shown in Table 4.

[0065] Table 3 Numbers, substitution groups, and physicochemical data of some compounds in the CAUZA-B series

[0066]

[0067] Table 4 H NMR and mass spectrometry data of some compounds in the CAUZL-B series

[0068]

[0069]

[0070]

[0071]

[0072] Example 3: Preparation of 1-10% CAUZA-A-01 emulsifiable concentrate

[0073] In a 100 mL volumetric flask, 1-10 g of compound CAUZA-A-01, 5-15 g of Tween 80, and 0.1-1 g of fatty alcohol polyoxyethylene ether were added, and then the volume was fixed with toluene to obtain an emulsifiable concentrate with a content of 1-10%.

[0074] Other emulsifiable concentrates of compounds with the general formula CAUZA-A and CAUZA-B can be prepared according to the above method.

[0075] Example 4: Preparation of wettable powder containing 15-50% of the compound CAUZA-A-01

[0076] Take 15-50 g of the compound CAUZA-A-01, 10-20 g of sodium dodecylbenzene sulfonate, and 30-75 g of white carbon black, mix and grind them to obtain a wettable powder with a content of 15-50%.

[0077] Other wettable powders of compounds of the general formula CAUZA-A and CAUZA-B can be prepared according to the above method.

[0078] Example 5. Determination of enzyme inhibition activity of compounds of the general formula CAUZA-A and CAUZA-B

[0079] Enzyme activity determination method: When measuring the inhibitory activity of OfChtI, 4-methylumbelliferyl N,N'-diacetyl-β-D-chitobioside (4-MethyluMbelliferyl N,N-diacetyl-β-D-chitobioside) was used as the test substrate of OfChtI, OfChtⅡ, and OfChi-h; p-nitrophenyl-2-acetylamino-2-deoxy-β-D-pyranoglucoside (pNP-β-GlcNAc) was used as the test substrate. The enzyme and enzyme activity buffer (20mM NaH2PO4, pH=6.5) were mixed in a 96-well plate to a final volume of 54μL, 6μL of 5mM pNP-β-GlcNAc was added to initiate the reaction, incubated at 30℃ for 20min, 60μL of 0.5M sodium carbonate was added to terminate the reaction, and the absorbance value was measured at 450nm.

[0080] Compound inhibitory activity assay method: The enzyme was incubated with different concentrations of inhibitors at room temperature for 10 min, and the enzyme activity was assayed at concentrations of 0.1 mM and 0.2 mM, respectively, using the above method. The inhibitory activity of some compounds on insect chitin degrading enzymes is shown in Tables 5 and 6 below.

[0081] Example 6: Determination of insecticidal activity of compounds of the general formula CAUZL-A and CAUZL-B

[0082] Determination method: The diamondback moth (Plutella xylostellaLinnaeus) and the Asian corn borer (Ostrinia furnacalis) were treated by the drip method and the leaf dipping method, and the results were checked after 3 days and 2 days respectively. The individuals that could not crawl normally after touching the insect body were considered dead, and their corrected mortality rate (%) was calculated. The toxicity of the agent was determined by comparing with the control agent. The insecticidal activity data of some compounds are shown in Tables 7 and 8.

[0083] The following targets were tested:

[0084] Plutella xylostella Linnaeus and Ostrinia furnacalis were purchased from commercial sources and raised indoors with corresponding feeds under the following conditions: room temperature (27±1)°C, humidity 80%, light intensity 2000 lux, and light duration 12 hours per day. Under indoor raising conditions, 3rd instar larvae with the same age, weight, and physiological condition were used for the drug activity screening test.

[0085] Table 5. Inhibition rate of some compounds of CAUZA-A series at 100 μM concentration (%)

[0086]

[0087] Table 6. Inhibition rate of some compounds of CAUZA-B series at 100 μM concentration (%)

[0088]

[0089] Table 7 Insecticidal activity of some compounds of CAUZL-A and CAUZL-B series against Plutella xylostella at a concentration of 500 mg / L (%)

[0090] Compound Lethality Rate (%) Compound Lethality Rate (%) CAUZA-A-01 46.52±3.5 CAUZA-B-01 57.46±5.01 CAUZA-A-02 48.15±6.44 CAUZA-B-02 53.60±6.03 CAUZA-A-03 74.07±6.44 CAUZA-B-03 7.20±2.00 CAUZA-A-04 59.26±6.45 CAUZA-B-04 76.80±6.40 CAUZA-A-05 69.14±7.73 CAUZA-B-05 65.20±8.20 CAUZA-A-06 83.93±7.72 CAUZA-B-06 82.60±8.15 CAUZA-A-07 70.37±6.43 CAUZA-B-07 61.33±7.00 CAUZA-A-08 38.89±7.84 CAUZA-B-08 38.13±6.41 CAUZA-A-09 63.13±4.94 CAUZA-B-09 76.80±7.59 CAUZA-A-10 72.33±7.82 CAUZA-B-10 30.40±5.39 CAUZA-A-11 77.78±6.51 CAUZA-B-11 26.54±8.20 CAUZA-A-12 74.09±6.415 CAUZA-B-12 71.00±5.60 CAUZA-A-13 72.22±7.8 CAUZA-B-13 65.20±7.00 CAUZA-A-14 60±7.8 CAUZA-B-14 76.80±8.02 CAUZA-A-15 74.41±6.35 - - Diflubenzuron 74.07±6.45 Diflubenzuron 76.96±3.35

[0091] Table 8 Insecticidal activity of some compounds of CAUZL-A and CAUZL-B series against Ostrinia nubilalis at a concentration of 500 mg / L (%)

[0092]

[0093] The present invention has been described in detail above. For those skilled in the art, without departing from the purpose and scope of the present invention, and without the need to carry out unnecessary experimental conditions, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. Although the present invention provides specific embodiments, it should be understood that the present invention can be further improved. In a word, according to the principles of the present invention, the application is intended to include any changes, uses or improvements to the present invention, including departure from the disclosed scope in the application, and changes made with conventional techniques known in the art.

Claims

1. A glycoside compound represented by formula (I) or formula (II) or a pharmaceutically acceptable salt, solvate or hydrate thereof: In formula (I) and formula (II), R is substituted or unsubstituted phenyl, benzyl or naphthyl; The substitution is mono- or poly-substitution; The substituents are each independently selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, and C1-C6 alkyl-substituted amino.

2. The compound according to claim 1, characterized in that The substituents are each independently selected from halogen, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, and C1-C6 alkyl-substituted amino.

3. A method for preparing the glycoside compound of formula (I) as claimed in claim 1, comprising the following steps: (1) reacting the compound represented by formula (III) with the compound represented by formula (IV) in an ethanol solvent to obtain a compound represented by formula (V); (2) deprotecting the compound represented by formula (V) under methanol-ammonia conditions to obtain the target compound represented by formula (I), The definition of R in formula (III) is the same as that of R in formula (I); The definition of R in formula (V) is the same as that of R in formula (I).

4. A method for preparing the glycoside compound represented by formula (II) according to claim 1, comprising the following steps: 1) reacting the compound represented by formula (V) in claim 3 with active manganese dioxide in chloroform to obtain the compound represented by formula (VI); The definition of R in formula (VI) is the same as that of R in formula (II); 2) Deprotecting the compound represented by formula (VI) under methanol-ammonia conditions to obtain the target compound represented by formula (II).

5. The compound represented by formula (VI) in claim 3 or the compound represented by formula (VI) in claim 4.

6. Use of the glycoside compound represented by formula (I) or (II) in claim 1 or its pharmaceutically acceptable salt, solvate, hydrate or composition comprising the same as an insect chitin degrading enzyme inhibitor in the preparation of insecticides.

7. The use according to claim 6, characterized in that: In the application, the insects are agricultural pests of the order Lepidoptera, including Plutella xylostella and Ostrinia nubilalis.

8. An insecticide comprising the glycoside compound represented by formula (I) or (II) in claim 1 or a pharmaceutically acceptable salt, solvate, hydrate thereof or a composition comprising the same.

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