Investigations into carbamate insecticide selectivity: I: Evaluation of potential selectophores

In an effort to improve the generally unfavorable mouse/housefly toxicity ratio of most carbamate insecticides, potential selectophores (nitrile, carbamoyl oxime, carboxylic ester, or amide) were incorporated into a series of phenyl N-methylcarbamates. In addition to the insect and mouse toxicity determinations, the anticholinesterase activity of these compounds was determined for purified housefly head acetylcholinesterase and bovine erythrocyte acetylcholinesterase. The presence of these functional groups, in general, did not give enhanced selectivity ratios and, in one case, (o-N-methylcarbamoyloxyiminomethylphenyl N-methylcarbamate), a very unfavorable selectivity ratio of <0.03 was obtained. A mechanism implicating a Beckmann rearrangement is advanced to rationalize the high rodenticidal activity of this molecule. In general, the carbamates showed poor insecticidal activity when applied alone to the housefly, but, when the flies were pretreated with piperonyl butoxide, the compounds were quite toxic. Finally, an explanation is derived which seeks to justify the inability of these potential selectophores to improve the mouse/housefly toxidity ratio.     

In vivo inhibition of esterase and acetylcholinesterase activities by profenofos treatments in the tobacco whitefly Bemisia tabaci (Genn.): Implications for routine biochemical monitoring of these enzymes

Insecticide-resistant populations of the tobacco whitefly, Bemisia tabaci (Genn.), lost over 90% of esterase activity and approximately 20% of acetylcholinesterase activity after treatment with 500 g/ha profenofos under field-simulated conditions. A more detailed study showed that esterase activity was completely inhibited within 2 days in insects confined on single cotton leaves treated with 50 g/ha profenofos (LD10) thereby preventing their discrimination from susceptibles which normally have low activity. However, esterase activity recovered sufficiently within 3 days when insects were transferred from the treated to untreated cotton leaves, thus enabling their correct classification as resistant. The inhibition/recovery profile for acetylcholinesterase closely followed that of the esterase system.     

Toxicological significance of acetylcholinesterase of the housefly thorax

Polyacrylamide gel electrophoresis of the 100,000g supernatant fraction of thoracic homogenates from the DDT/S housefly strain separated five bands having acetylcholinesterase activity. Ferguson plots indicated that the three bands of slowest mobility constituted members of the heterologous-size isomer family described for the head enzyme. The two faster migrating bands appeared as electrophoretic artifacts of juxtaposition with a 3.6 S nonacetylcholinesterase component; in the absence of juxtaposition, a single form was characterized with identical molecular properties to the 5.3 S fundamental unit of head acetylcholinesterase. The kinetic properties of isolated forms were consonant with the molecular data and no major differences in K_m values for acetylthiocholine were discovered between the thoracic and head enzymes. We suggest the preferential in vivo inhibition of thoracic acetylcholinesterase by organophosphate pesticides is unlikely the consequence of biochemical differences between the enzymes from the two regions.     

Cross‐resistance and possible mechanisms of chlorpyrifos resistance in Laodelphax striatellus (Fallén)

BACKGROUND: Laodelphax striatellus (Fallén) is a major pest of cultivated rice and is commonly controlled in China with the organophosphate insecticides. To develop a better resistance management strategy, a chlorpyrifos‐resistant strain of L. striatellus was selected in the laboratory, and its cross‐resistance to other insecticides and possible mechanisms of the chlorpyrifos resistance were investigated. RESULTS: After 25 generations of selection with chlorpyrifos, the selected strain of L. striatellus developed 188‐fold resistance to chlorpyrifos in comparison with the susceptible strain, and showed 14‐ and 1.6‐fold cross‐resistance to dichlorvos and thiamethoxam respectively. There was no apparent cross‐resistance to abamectin. Chlorpyrifos was synergised by the inhibitor triphenyl phosphate; the carboxylesterase synergistic ratio was 3.8 for the selected strain, but only 0.92 for the susceptible strain. The carboxylesterase activity of the selected strain was approximately 4 times that of the susceptible strain, whereas there was no significant change in the activities of alkaline phosphatase, acid phosphatase, glutathione S‐transferase and cytochrome P450 monooxygenase between the strains. The Michaelis constant of acetylcholinesterase, maximum velocity of acetylcholinesterase and median inhibitory concentration of chlorpyrifos‐oxon on acetylcholinesterase were 1.7, 2.5 and 5 times higher respectively in the selected strain. CONCLUSION: The high cross‐resistance to the organophosphate dichlorvos in the chlorpyrifos‐resistant strain suggests that other non‐organophosphate insecticides would be necessary to counter resistance, should it arise in the field. Enhanced activities of carboxylesterase and the acetylcholinesterase insensitivity appear to be important mechanisms for chlorpyrifos resistance in L. striatellus. Copyright © 2010 Society of Chemical Industry     

球孢白僵菌次生代谢产物对南方根结线虫的作用机制初探

为明确球孢白僵菌Beauveria bassiana对南方根结线虫Meloidogyne incognita的作用机理,从线虫的活动频率、呼吸强度、体液渗透压、总糖含量、可溶性蛋白含量以及乙酰胆碱酯酶(AChE)活性几方面测定了球孢白僵菌Snef23菌株次生代谢产物对南方根结线虫2龄幼虫(J2)的影响。结果表明:Snef23次生代谢产物可抑制南方根结线虫J2的活动频率和呼吸作用,降低其乙酰胆碱酯酶活性,增强虫体内容物的渗漏,干扰J2体内糖和可溶性蛋白的代谢,从而达到毒杀致死作用。研究结果为探明球孢白僵菌杀线虫的作用机制提供了理论依据,可为更好地利用球孢白僵菌及其他天然产物防控植物的线虫病害提供参考。     

Use of novel approaches for designing safer drugs in insecticide research: Soft alkylating agents

Based on the analogy between anticancer drugs and insect chemosterliants, both of them acting by virtue of their alkylating capacity, chloromethyl carboxylates (described by Bodor and co-workers as potential anticancer drugs of the soft alkylating agent type) including the chloromethyl ester analogues of the juvenoids hydroprene and methoprene were prepared and tested for chemosterilant and morphogenetic juvenile hormone activity. While the chloromethyl esters of some simple carboxylic acids were inactive in this respect, the juvenoid chloromethyl esters revealed both chemosterilant and morphogenetic activity against Dysdercus cingulatus comparable with that of the known juvenoids hydroprene and methoprene. At the same time their activity against Pieris brassicae showed a sharp drop compared to the ‘parent’ compounds. The results suggest a species-dependent rate of enzymatic hydrolysis of the chloromethyl ester group as a possible reason for their selective action. Thus, the chloromethyl ester group might function as a ‘selectivity factor’).     

Phosphinate inhibition studies of cholinesterases

The kinetic constants, K_d, k₂, and k_i, were determined for the inhibition by 4-nitro-phenyl methyl(phenyl)phosphinate of three cholinesterases: butyrylcholinesterase, bovine erythrocyte acetylcholinesterase and eel acetylcholinesterase. Stopped-flow kinetic evaluations and automated data acquisition and processing were employed. A broad range in affinity for the phosphinate inhibitor was observed as reflected by the binding constants, K_d. A similar wide range in the k₂ values for the unimolecular inhibition step was obtained. The net bimolecular rate constants, ki, indicate equal overall reactivity for butyrylcholinesterase and eel acetylcholinesterase with a smaller inhibition rate constant for bovine erythrocyte acetylcholinesterase.     

The antifungal activity of alkyl benzimidazol-2-ylcarbamates and related compounds

The fungistatic activity against Penicillium digitatum and Diplodia natalensis decreased slightly in ascending a homologous series of alkyl esters of benzimidazol-2-ylcarbamic acid from the methyl ester (carbendazim) to the pentyl ester; the hexyl and octyl esters were inactive. 2-(Acylamino)benzimidazoles were slightly less active than the analogous alkyl benzimidazol-2-ylcarbamates. Introduction of a methylene bridge between the benzimidazole ring and the 2-methoxycarbonylamino group abolished antifungal activity. Methylation of either the carbamate nitrogen or an imidazole nitrogen of carbendazim produced inactive compounds. Replacement of the benzimidazole ring of carbendazim with various other ring systems was accompanied by marked reduction in antifungal activity.     

Stereochemical basis for the insecticidal activity of carbamoylated and acylated pyrazolines

Methyl 3-(4-chlorophenyl)-1-[N-(4-chlorophenyl)carbamoyl]-4-methyl-2-pyrazoline-4-carboxylate was converted to corresponding (1R)- and (1S)-phenethyl esters via its carboxylic acid and acid chloride at the C-4 atom to separate the diastereomers. Their configurations were confirmed by X-ray analysis. Both isomers of the (1R)methylbenzyl ester were subjected to transesterification with sodium methoxide to obtain enantiomers of the starting methyl ester. Their insecticidal activity was measured against American cockroaches (Periplaneta americana (L.)) by injection and against house flies (Musca domestica L.) by topical application under various synergistic conditions with metabolic inhibitors. The activity values of the four α-methylbenzyl esters and the R-isomer of the starting methyl ester were similar. The S-enantiomer of the methyl ester was about 10 and 100 times more active than the R-isomer against the cockroach and the fly, respectively. Some N-arylacetyl and N-aryloxyacetyl derivatives of the starting N-(4-chlorophenyl)carbamoyl compound gave very low activity. Conformation-energy profiles for some compounds suggested that the conformation of substituents on the N-1 atom in the pyrazoline ring has a specific role for the potential insecticidal effects.     

The role of B‐type esterases in conferring insecticide resistance in the tobacco whitefly,Bemisia tabaci (Genn)

Separation of non‐specific esterases on electrophoretic gels has played a key role in distinguishing between races or biotypes of the tobacco whitefly, Bemisia tabaci. One intensively staining esterase in particular (termed E_0.14) has assumed significance as a diagnostic of B‐type whiteflies (aka Bemisia argentifolii), despite any knowledge of its biological function. In this study, a whitefly strain (B‐Null) homozygous for a null allele at the E_0.14 locus that had been isolated from a B‐type population was used to demonstrate a significant role for E_0.14 in resistance of B‐type populations to pyrethroids but not to organophosphates (OPs). Bioassays with pyrethroids, following pre‐treatment with sub‐lethal doses of the OP profenofos (to inhibit esterase activity), coupled with metabolism studies with radiolabelled permethrin, supported the conclusion that pyrethroid resistance in a range of B‐type strains expressing E_0.14 was primarily due to increased ester hydrolysis. In the same strains, OP resistance appeared to be predominantly conferred by a modification to the target‐site enzyme acetylcholinesterase. © 2000 Society of Chemical Industry     

Acetylcholinesterase inhibition by nootkatone and carvacrol in arthropods

The essential oils from many botanicals have been screened for insecticidal activity. Two constituents of the Alaskan yellow cedar tree, the monoterpenoid carvacrol and the sesquiterpenoid nootkatone, both are toxic against several arthropods. The mode of action through which nootkatone and carvacrol exert their insecticidal activity remains uncertain. It has been hypothesized that they may inhibit acetylcholinesterase enzyme activity. The degree of acetylcholinesterase inhibition of carvacrol and nootkatone was compared to that of carbaryl, a known acetylcholinesterase inhibitor, in the house fly (Musca domestica), yellow fever mosquito (Aedes aegypti), American dog tick (Dermacentor variabilis) and American cockroach (Periplaneta americana). The concentration of carbaryl, at which 50% of the acetylcholinesterase activity was inhibited (IC₅₀), was less than 2 μM in all four arthropod models. Carvacrol was observed to cause slight inhibition of the acetylcholinesterase enzyme in house flies, ticks and cockroaches, but it did not inhibit the mosquito acetylcholinesterase enzyme. Nootkatone did not inhibit the acetylcholinesterase enzyme in any of the four arthropod models tested. From this study, we conclude that the acetylcholinesterase inhibition is not likely the primary mode of action for insecticidal activity by nootkatone or carvacrol.     

Comparative properties of N-acetyl derivatives of oxime N-methylcarbamates and aryl N-methylcarbamates as insecticides and acetylcholinesterase inhibitors

N-acylation of aryl N-methylcarbamates is known to reduce their mammalian toxicity considerably without adversely affecting their insecticidal activity. It has now been found that N-acylation of several insecticidal oxime carbamates results in loss of toxicity both to insects and mammals. Kinetic data for base-catalysed solvolysis and inhibition of acetylcholinesterase, together with published work on metabolism, are combined to provide a rationale for this unexpected observation and to account for the selective toxicity of aryl N-acetyl-N-methylcarbamates.     

阿魏酸衍生物的合成及其除草活性测定

为从天然产物中获取新的除草活性成分,通过化学合成的方法对分离自瓜果腐霉Pythium aphanidermatum代谢产物中的阿魏酸进行了结构改造,并采用小杯法,以马唐、马齿苋和播娘蒿为供试杂草对所得化合物进行了除草活性测定。结果显示:所合成的4-羟基-3-甲氧基肉桂酸乙酯、4-羟基-3-甲氧基肉桂酸邻氯苯胺、4-羟基-3-甲氧基肉桂酸酰肼、4-羟基-3-甲氧基肉桂酸叔丁酯和4-乙酰氧基-3-甲氧基肉桂酸5种化合物的回收率分别为96.81%、97.39%、82.20%、75.67%和98.29%。4-羟基-3-甲氧基肉桂酸酰肼和4-乙酰氧基-3-甲氧基肉桂酸对马齿苋胚根的抑制活性最强,IC50分别为87.50 mg/L和74.86 mg/L,其余3种活性相对较弱;4-羟基-3-甲氧基肉桂酸乙酯和4-乙酰氧基-3-甲氧基肉桂酸分别在500 mg/L和1 000 mg/L浓度下可完全抑制播娘蒿和马齿苋胚根、芽的生长,而4-羟基-3-甲氧基肉桂酸邻氯苯胺则在1 000 mg/L浓度下可完全抑制播娘蒿胚根的生长。表明在阿魏酸分子中的不同位点引入适当的基团结构可以提高或者降低阿魏酸分子的除草活性,并获得新的除草活性物质。     

Is acetyl/butyrylcholine specificity a marker for insecticide‐resistance mutations in insect acetylcholinesterase?

Substrate specificity has been widely studied in vertebrate cholinesterases and it has been shown that two phenylalanines in the acyl pocket of acetylcholinesterase govern the acceptance of the acetyl/butyryl moiety of the choline esters. As an insecticide‐resistance mutation has been evidenced in the acyl pocket of Drosophila melanogaster and Musca domestica acetylcholinesterase we investigated the possibility of linking changes in acetyl/butyrylthiocholine specificity with mutations in insect acetylcholinesterase. We thus analyzed the effect of 28 mutations in Drosophila enzyme on acetyl/butyrylthiocholine, N‐methyl/N‐propyl‐carbamates and ethyl/methyl‐paraoxon preference. It appeared that the highest changes on acetyl/butyrylthiocholine and N‐propyl/N‐methyl‐carbamates preference were due to mutations in the acyl pocket. Nevertheless, other insecticide‐resistance mutations, not located in the acyl pocket, also modified these substrate preferences. Moreover, the effect of mutations in the acyl pocket was hidden when some other insecticide‐resistance mutations were combined in the enzyme. Consequently, acetyl/butyrylthiocholine preference alteration cannot be used as a marker to localize a mutation in the insect AChE. © 2000 Society of Chemical Industry     

寄主植物对桃蚜羧酸酯酶和乙酰胆碱酯酶的诱导作用

在１９９５～１９９６年研究了寄主植物对桃蚜［Ｍｙｚｕｓｐｅｒｓｉｃａｅ（Ｓｕｌｚｅｒ）］羧酸酯酶（ＣａｒＥ）和乙酰胆碱酯酶（ＡＣｈＥ）的诱导作用。在试验的甘蓝、茄子和桃树３种寄主植物中，取食甘蓝的桃蚜种群ＣａｒＥ和ＡＣｈＥ活性最高，取食茄子和桃树的桃蚜种群ＣａｒＥ活性没有明显不同，而ＡＣｈＥ活性取食茄子的桃蚜种群明显高于取食桃树的种群。ＣａｒＥ与底物的亲和力是桃树＞茄子＞甘蓝，而ＡＣｈＥ与底物的亲和力则是甘蓝＞茄子＞桃树。ＡＣｈＥ与毒扁豆碱的双分子速率常数（Ｋｉ）值大小顺序为甘蓝＞桃树＞茄子     

Mechanisms conferring resistance of Western flower thrips to bendiocarb

Mechanisms associated with bendiocarb resistance were examined in two strains of western flower thrips, Frankliniella occidentalis (Pergande), that differed in their susceptibility to this carbamate by 13.6-fold. No appreciable differences between the two strains in [¹⁴C] bendiocarb penetration and excretion were detected; however, bendiocarb was metabolised substantially faster by the resistant KCM thrips than by the more susceptible UMC thrips. No appreciable difference between the two strains was found in the sensitivity of acetylcholinesterase activity to inhibition by bendiocarb. It was concluded that bendiocarb resistance in KCM western flower thrips was due to enhanced metabolism that probably was mainly oxidative in nature.     

白背飞虱对马拉硫磷和异丙威的抗性机制

测定了白背飞虱不同品系体内与抗药性相关酶系如酯酶、羧酸酯酶、谷膦甘肽S-转移酶和乙酰胆碱酯酶等活性。与敏感品系S相比，抗马拉硫磷品系顺具高水平羧酸酯酶活性。酶反应动力学试验表明，羧跋酯酶的质与量均发生改变；等电聚焦电泳显示与pI5．26～5．84酯酶扩增有关。耐弄丙威品系IT乙酰胆碱酯酶活性相对较低，而药剂对其I50则明显高于S。不同品系的谷胱甘肽S—转移酶活性无显著差弄。作者认为，白背飞虱对马拉硫磷和异丙威产生抗性分别与羧酯酯酶活性增高和乙酰胆碱酯酶对药剂敏感性降低有关。     

Dimethoxypyrimidines as Novel Herbicides. Part 2. Synthesis and Herbicidal Activity of O-Pyrimidinylsalicylates and Analogues

A new series of the O-pyrimidinylsalicylates was synthesized and their herbicidal activity was examined. Some of these compounds showed very strong herbicidal activity under pre- and post-emergent treatment conditions against various kinds of grass and broadleaf weeds. Among these compounds, O-(4, 6-dimethoxypyrimidin-2-yl) salicylic acid and its methyl ester were found to exhibit the highest activity. The herbicidal symptoms observed after the treatments included early cessation of plant growth followed by chlorosis, necrosis and plant death. The symptoms were similar to those caused by sulfonylureas and imidazolinones, which inhibit branched-chain amino acid biosynthesis.     

Pyrethroid insecticides derived from substituted biphenyl-3-ylmethanols

The synthesis of a series of mono- and disubstituted biphenyl-3-ylmethyl esters of 3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropanecarboxylic acid is described. The bioactivity of these compounds against Spodoptera eridania, Epilachna varivestis, Oncopeltus fasciatus, Acrythosiphon pisum and Tetranychus urticae is presented and discussed. Substitution of fluorine, chlorine and methyl groups in the 2-position of the biphenyl ring generally led to an increase in activity over the unsubstituted parent biphenyl ester. In addition, pyrethroid esters derived from these 2-substituted biphenyl-3-ylmethanols appeared to have a broader spectrum of activity than ‘classical’ pyrethroids. For example, the (1RS)-cis-3-(2,2-dichlorovinyl)-2, 2-dimethylcyclopro-panecarboxylic acid ester of 2-methylbiphenyl-3-ylmethanol was acaricidal, while maintaining a level of activity against other insects that was equal to or greater than cis-permethiin. Biological data on other esters of this novel alcohol are also presented.     

Species differences in the in vitro inhibition of brain acetylcholinesterase and carboxylesterase by mipafox,paraoxon, and soman

Acute intraperitoneal toxicity of mipafox, paraoxon, and soman was highest in chicken, followed by rat, and lowest in frog. Species differences in organophosphorus toxicity were not related to differences in the specific activities of either acetylcholinesterase or carboxylesterase in brain. The sensitivity to inhibition of brain acetylcholinesterase in vitro by the organophosphorus compounds was closely related to the susceptibility of the species to acute organophosphorus poisoning. Both the acute toxicity and the sensitivity of brain acetylcholinesterase to inhibition in vitro by organophosphorus compounds in all the species were in the following order of increasing activity: mipafox < paraoxon < soman. The sensitivity of brain carboxylesterase to inhibition by the organophosphorus compounds were less than that of acetylcholinesterase and it could not be related to species susceptibility to acute organophosphorus toxicity. Paraoxon-insensitive phenyl valerate hydrolase in chicken brain was more sensitive to inhibition by mipafox and soman compared to rat; in chicken the sensitivity of paraoxon-insensitive phenyl valerate hydrolase to inhibition by soman was 9000 times more than that by mipafox, while in rat it was 100 times more. Frog brain had no paraoxon-insensitive phenyl valerate hydrolase activity. No evidence of age dependence was noticed in the specific activities of brain acetylcholinesterase, carboxylesterase, neurotoxic esterase, and paraoxon-insensitive phenyl valerate hydrolase or in the sensitivity of these enzymes to in vitro inhibition by organophosphorus compounds in both rats and chickens.