Organosilane insecticides. Part I: Biological and physical effects of isosteric replacement of silicon for carbon in etofenprox and MTI-800

Substitution of silicon for carbon was explored in etofenprox and MTI-800, primarily to establish the suitability of silicon for pesticide construction and to probe steric effects. It was shown that silicon is an effective building block for insecticides with subtle qualitative and quantitative differences relative to the carbon analogs. The silanes were slightly less toxic to insects than the carbon analogs (relative potency of 0.2-0.6) but the silane analog of MTI-800 was considerably less toxic to fish (0% mortality at ≥50 mg liter^?1) than MTI-800 itself (LC₅₀ = 3 mg liter^?1). The mode of action of both carbon and silicon compounds was similar in the intact, electrode-implanted cockroach and involved repetitive firing of a sensory nerve; potency measurements were also made using an in-vitro nerve assay. The possible metabolic and physicochemical contributions to the observed insecticide potencies were explored using synergism studies, physical chemistry measurements and quantum mechanical calculations.     

Inhibitory effects of BHC isomers on Na+-K+-ATPase,yeast growth,and nerve conduction

The biological effects of benzene hexachloride (BHC, 1,2,3,4,5,6-hexachlorocyclohexane) isomers, such as toxicity against yeasts, inhibition of beef brain Na⁺-K⁺-ATPase, and blocking action on conduction in cockroach nerve, were determined and compared with those shown by homologous alcohols and other molecules. Each type of biological activity correlated well with the physicochemical properties of the test compounds such as hydrophobicity (as defined by their partition coefficients in 1-octanol/H₂O). The insecticidal action of γ-BHC showed little correspondence to Na⁺-K⁺-ATPase inhibition or to nerve blocking in insects.     

Interaction of tralomethrin,tralocythrin, and related pyrethroids in Na+ channels of insect and mammalian neuronal cells

The neurotoxic activity of the tetrahalogenated pyrethroids, tralomethrin and tralocythrin, which are dibromo adducts of deltamethrin and RU 24501, respectively, and RU 27218, which is a dichloro adduct of RU 24501, have been studied on two different excitable membranes using in vitro methods: insect axonal membranes and mammalian neuroblastoma cells in culture. The results show that molecules whose chrysanthemic acid chain at C₃ is saturated by halogen addition have intrinsic toxicity on the sodium channels of nerve membranes. They increase the amplitude of negative after-potential, depolarize the cockroach axonal membrane, and stabilize an open conformation of voltage-dependent Na⁺ channels in mouse neuroblastoma cells, thereby producing a massive uptakeof Na⁺ by the cells. All effects were observed in the absence of metabolic transformation of the compounds tested. In light of these experiments, it is concluded that tralomethrin and tralocythrin cannot be considered as propyrethroids.     

Neurophysiological effects of insecticides on the labellar taste receptors of Lucilia cuprina Wied

The effect of insecticide concentration on the nerve impulses at the labellar receptors of the blowfly Lucilia cuprina Wied was tested and found to be significant for the DDT type of insecticide, but not for the DDT-pyrethroid type. The experiments were performed at three different times of the day, but time of day had no effect on the multiplicity indices calculated from nerve spikes induced by the compounds. No effects on the nerve response were elicited by the synergistic action of sesamex.     

Effects of deltamethrin on ventral nerve cord activity in the cockroach

The effects of varying concentrations of deltamethrin on a number of the parameters of electrophysiological activity in the cockroach ventral nerve cord have been studied. Deltamethrin, at concentrations greater than 100 nM, caused repetitive firing in the central nervous system (CNS), prior to conduction block, the effect getting faster as the concentration was increased. Whilst 10-nM deltamethrin eventually caused conduction block with no apparent increase in the level of CNS activity, it induced a gradual decrease in the amplitudes of all spike activity. Deltamethrin at 10μM induced a significant increase in the latency of electrically evoked responses, and this was attributed to a synaptic mechanism. It was shown for the first time that 10-μM deltamethrin causes a significant elevation of the extra-axonal K⁺ activity; the possible consequences of this are discussed.     

Two classes of pyrethroid action in the cockroach

Pyrethroids are divided into two classes (Types I and II) based on their effects on the cercal sensory nerves recorded in vivo and in vitro and on the symptomology they produce in dosed cockroaches, Periplaneta americana. Type I compounds include pyrethrins, S-bioallethrin, [1R,cis]resmethrin, kadethrin, the 1R,trans and 1R,cis isomers of tetramethrin, phenothrin, and permethrin, and an oxime O-phenoxybenzyl ether. Electrophysiological recordings from dosed individuals reveal trains of cercal sensory spikes and sometimes also spike trains from the cercal motor nerves and in the CNS. Low concentrations of these pyrethroids act in vitro to induce repetitive firing in a cercal sensory nerve following a single electrical stimulus. This in vitro measurement, standardized for evaluating structure-activity relationships, shows that only 1R, insecticidal isomers are highly effective neurotoxins. The most potent compounds on the isolated nerve are [1R,trans]- and [1R,cis]tetramethrin, each active at 3 × 10^?13M. The poisoning symptoms of Type I compounds are restlessness, incoordination, hyperactivity, prostration, and paralysis. Type II compounds include [1R,cis,αS]- and [1R,trans,αS]cypermethrin, deltamethrin, and [S,S]fenvalerate. These α-cyanophenoxybenzyl pyrethroids do not induce repetitive firing in the cercal sensory nerves either in vivo or in vitro; moreover, they cause different symptoms, including a pronounced convulsive phase. Two other pyrethroids with an α-cyano substituent, i.e., fenpropathrin and an oxime O-α-cyanophenoxybenzyl ether, are classified as Type I based on their action on a cercal sensory nerve but the symptoms with these compounds resemble Type II. The two classes of pyrethroid action evident with the cockroach are discussed relative to their neurophysiological effects and symptomology in other organisms.     

Picrotoxinin receptor in the central nervous system of the American cockroach: Its role in the action of cyclodiene-type insecticides

The nature of the picrotoxinin receptor was studied using the central nervous system (CNS) of the American cockroach. It first was confirmed by using an electrophysiological technique that the abdominal nerve cord of the American cockroach was sensitive to picrotoxinin. By using a $\text{[}{}^3\text{H]α-dihydropicrotoxinin}$ binding test it was determined that the picrotoxinin receptor in CNS of this insect had a higher affinity toward picrotoxinin and heptachlor epoxide than the corresponding receptor in the rat brain. Also, the cockroach brain preparation had a higher percentage of specific binding in the total binding, making this material suitable for receptor studies. By using a sucrose density centrifugation technique, it was determined that the fraction sedimented at the interphase of 1.0 to 1.2 M sucrose at 100,000g contained the highest level of specific binding site. The receptor showed a sensitivity to all insecticidal cyclodienes tested, namely photodieldrin, oxychlordane, endrin, heptachlor epoxide, γ-chlordane, dieldrin, aldrin, heptachlor, and isodrin (expressed in the order of potency). Among four BHC isomers, the γ-isomer showed the highest potency to bind with this receptor.     

Structure-related effects of pyrethroid insecticides on the lateral-line sense organ and on peripheral nerves of the clawed frog, Xenopus laevis

The effects of seven different pyrethroid insecticides on the lateral-line sense organ and on peripheral nerves of the clawed frog, Xenopus laevis, were investigated by means of electrophysiological methods. The results show that two classes of pyrethroid can be clearly distinguished. (i) Pyrethroids without an α-cyano group (permethrin, cismethrin, and bioresmethrin). These noncyano pyrethroids induce short trains of nerve impulses in the lateral-line sense organ. In peripheral nerve branches they induce a depolarizing afterpotential and repetitive firing. These effects are very similar to those previously reported for allethrin. (ii) Pyrethroids with an α-cyano-3-phenoxybenzyl alcohol (cypermethrin, fenpropathrin, deltamethrin, and fenvalerate). In the lateral-line sense organ these α-cyano pyrethroids induce very long trains of nerve impulses which may last for seconds and may contain hundreds or even thousands of impulses. The α-cyano compounds do not cause repetitive activity in peripheral nerves. Instead they induce a quickly reversible, stimulus frequency-dependent suppression of the action potential. Since the chemical structure of cypermethrin differs from that of permethrin only in the α-cyano group and because all four α-cyano compounds act in a very similar way, it is concluded that the α-cyano substituent is responsible for the large differences in neurotoxic effects. In the lateral-line sense organ the duration of nerve impulse trains induced by the noncyano as well as the α-cyano pyrethroids increases dramatically when the temperature is lowered. Further, in sensory fibers the effects of both classes of pyrethroid on the nerve action potential are more pronounced compared to their effects on motor fibers. It is argued that the different neurotoxic effects reported here originate from a common mechanism of action of pyrethroids, which is a prolongation of the transient increase of sodium permeability of the nerve membrane associated with excitation.It is concluded that the sodium channel in the nerve membrane is the major target site of noncyano and α-cyano pyrethroids.     

The importance of nerve terminal depolarization in pyrethroid poisoning of insects

The effects of several pyrethroids and DDT were examined on neuromuscular preparations from larvae of Musca domestica, Culex quinquefasciatus, Anopheles stephensi, and Trichoplusia ni, in order to determine the importance of the type I effect (repetitive firing) and the type II effect (increased miniature excitatory postsynaptic potential (mepsp) rate, indicating nerve terminal depolarization) in the poisoning process. α-Cyano pyrethroids were very potent in increasing mepsp rate, and although DDT and non-α-cyano compounds were more potent at inducing repetitive firing, all these compounds increased mepsp rate at higher concentrations. A variety of evidence showed that the mepsp rate-increasing activity could be an important factor in toxicity: among all of the compounds, there was a positive correlation between toxicity and mepsp rate-increasing activity; mepsp rate-increasing activity had a negative temperature dependence; nerves of kdr larvae in vitro were resistant to the mepsp rate-increasing activity, commensurate with resistance level; and finally, increased mepsp rate and neuromuscular block were observed in poisoned insects, associated with paralysis. Among type I compounds, repetitive firing activity was correlated negatively with toxicity, but positively with knockdown activity.     

Quantitative structure-activity studies of substituted benzyl chrysanthemates: 1. Correlations between symptomatic and neurophysiological activities against American cockroaches

A number of substituted benzyl (1R)-trans-chrysanthemates and related compounds were synthesized. Their symptomatic activities in terms of levels which induce convulsions as well as cause death in American cockroaches were determined by injection with and without application of synergists as inhibitors of metabolism. The neuroexcitatory and neuroblocking activities were also determined in terms of minimum effective concentrations to induce repetitive train of impulses and conduction blockage, respectively, to central nerve cords excised from the cockroaches and immersed in Ringer's solution. Correlations between symptomatic and neurophysiological activities were analyzed quantitatively with the aid of molecular hydrophobicity parameter and regression analysis. Each symptomatic activity from which the effect of metabolism is eliminated was found to be analyzable by means of a linear combination of indices for two types of neurophysiological activity when the transport factor is separated by using the hydrophobicity parameter. A closer correlation was found between neuroblocking activity and the “convulsive” effect than between neuroexcitatory activity and the “convulsive” effect, whereas both neurophysiological effects operate together on the cockroaches resulting in paralysis and death.     

A comparison of DDT and its biodegradable analogues tested on ATPase enzymes in cockroach

Analogues of DDT (ethoxymethyl and methoxymethio derivatives) compared with DDT for their inhibitory action on the ATPase system from tissues of the cockroach, Periplaneta americana show similar, but less inhibitory effects. The mitochondrial (oligomycin-sensitive) Mg²⁺ ATPase activity from coxal muscle preparations was more sensitive to DDT than the two analogues; whereas, the muscle and nerve cord homogenates showed about equal sensitivity to the biodegradable analogues. The mitochondrial Mg²⁺ ATPase from nerve cord preparation was more sensitive to the three compounds than the Na⁺K⁺ ATPase activity. The significance of these results in relation to recent reports on the effect of DDT on Na⁺K⁺ ATPase is discussed.     

Temperature dependence of allethrin-induced repetitive discharges in nerves

The action of allethrin in inducing repetitive discharges by a single stimulus has been studied with squid giant axons at various temperatures. Repetitive discharges are evoked in the allethrin-poisoned axon at optimal temperatures ranging from 20 to 28°C. A single action potential is produced below and above this temperature range. The negative (depolarizing) after-potential is increased markedly by allethrin, and it exceeds the threshold membrane potential for excitation only at the optimal temperatures, thereby accounting for the temperature dependence of repetitive responsiveness.     

The effects of dieldrin and isomeric aldrin diols on synaptic transmission in the American cockroach and their relevance to the dieldrin poisoning syndrome

Dieldrin and two of its metabolites, 6,7-trans-dihydroaldrindiol, and 6,7-cis-dihydroaldrindiol, were studied with regard to their toxicity to the American cockroach, effects on ganglia of the ventral nerve cord, and penetration into the ventral nerve cord of poisoned cockroaches. An approximate LD₅₀ for injected doses of dieldrin was 0.45 mg/kg. After injection at 115 mg/kg, the trans isomer of aldrin diol caused about 70%, and the cis isomer about 50% mortality. Injected doses of 40 mg/kg of the three compounds appeared in the ventral nerve cord to the extent of 0.13–0.26% of the doses. Dieldrin was more potent, but slower acting than the diols in causing synaptic after-discharge and elevated spontaneous activity in isolated nerve cords. The results are discussed in relation to other studies on these compounds. It was concluded that, in the American cockroach, dieldrin, rather than either of the diols, is the insecticidal agent in dieldrin poisoning, and that metabolic conversion of dieldrin to the cis and/or trans aldrindiol constitutes a detoxification.     

Effects of organosilicon pyrethroid‐like insecticides on nerve preparations of American cockroaches and crayfish

Quaternary organosilicon pyrethroid‐like ethers (five compounds) and alkanes (three compounds) were used for neurophysiological tests. Their activities in inducing repetitive firing in the central nervous cord of the American cockroach (Periplaneta americana) were evaluated by an extra‐cellular recording method. The ethers were more active than the corresponding alkanes. The ability of the compounds to cause conduction blockage was also measured using the same nerve preparations, but the effects were too weak to allow definitive activity values to be determined. The compounds prolonged the sodium tail‐current in the crayfish giant axon under voltage clamp conditions. The rate of decay of the tail‐current changed in parallel with the reported insecticidal activity against P americana. These findings indicated that tail‐current activity was the most useful nerve parameter in predicting insecticidal activity. Regression analysis of the numerical data together with those reported for other alkanes revealed that the higher the tail‐current activity, the higher the insecticidal activity when a structural feature and the hydrophobicity were considered separately. The insecticidal activity of the ethers was about seven‐fold higher than that of the alkanes with equivalent tail‐current activity and hydrophobicity. Variations in insecticidal activity were parabolically correlated with the hydrophobicity [(log P)_opt = 9.1] when other factors were similar. © 2001 Society of Chemical Industry     

Dibromo addition products of (1R)-cis-permethrin and deltamethrin; evidence that debromination is an obligatory requirement for biological activity

3-Phenoxybenzyl (1R-)-cis-3-(1,2-dibromo-2, 2-dichloroethyl)-2, 2-dimethylcyclopropanecarboxylate [dibromo-(1R)-cis-permethrin] was >100 times less active than (1R)-cis-permethrin in causing repetitive firing in a cockroach cereal sensory nerve in vitro, although its toxicity, symptomology and nerve effects in vivo were comparable to the parent compound. Treatment of the in-vitro preparation with glutathione significantly increased the effectiveness of dibromo-(1R)-cis-permethrin in a concentration-dependent manner, while having no effect on the action of (1R)-cis-permethrin. Topical treatment of the cockroach with dibromo-(1R)-cispermethrin gave a similar time of onset of symptoms to that caused by (1R)-cis-permethrin (∼2 min). Pretreatment of the cockroach with N-ethylmaleimide (NEM) significantly (P<0.05) increased dose-dependently the latency of signs of poisoning caused by dibromo-(1R)-cis-permethrin, but had no significant effect on the latency following (1R)-cis-permethrin treatment. Deltamethrin and tralomethrin, applied to the cockroach at LD₉₅ doses of 0.03 and 0.07 m̈g g⁻¹ respectively, also had similar latencies for the development of symptoms. Moreover, pretreatment with NEM delayed the time of onset of symptoms for tralomethrin (P<0.02) but not for deltamethrin. The results strongly suggest that debromination is an obligatory requirement for the biological activity of dibromo-(1R)-cis-permethrin and tralomethrin.     

Pyrethroid resistance in a strain of Spodoptera littoralis is correlated with decreased sensitivity of the CNS in vitro

The effects of permethrin and cypermethrin on the isolated abdominal nerve cord of insecticide-resistant [R] and -susceptible [S] strains of Spodoptera littoralis larvae have been studied. Above ca. 19°C, permethrin at 10^?7M caused a prolonged spike train to follow electrical stimulation of the nerve cord. The time of onset of this repetitive firing was significantly greater for the [R] strain. Moreover, cypermethrin, to which this strain shows negligible resistance, did not cause such repetitive discharges. Thus, resistance to permethrin but not to cypermethrin appears to be based on a qualitative difference between the pyrethroids. Nerve blockage by the two pyrethroids was also investigated, with particular reference to temperature. Once again, differences were apparent: when considered relative to untreated controls, permethrin caused quicker nerve blockage as temperature was reduced whereas the blocking action of cypermethrin was not affected by temperature. However, the times taken to cause nerve blockage by permethrin in [R] and [S] larvae were not significantly different, making it unlikely that nerve blockage plays a major role in this resistance. Two methods were employed to reduce the resistance factor in vitro. The synergist dodecyl imidazole failed to significantly reduce the time taken for permethrin to cause either repetitive firing or nerve blockage. However, reducing the calcium concentration in the saline did significantly reduce the latency of repetitive firing caused by permethrin in [R] larvae, thus increasing the nerve sensitivity to approximately the same level as normal calcium, [S] insects.     

The effects of DDT analogs upon potassium conductance in synthetic membranes

The effects of p,p′-DDT and 13 analogs were studied upon the K⁺ conductance which valinomycin induces in a lecithin-octane bilayer. Eight compounds decreased conductance, 4 relatively polar analogs increased conductance, and one had no effect. A partial correlation of these variations with physiological effects upon cockroach nerve and crayfish giant axon was found. Evidence was presented that the effect on the bilayer was due to an effect of the compounds on the fluidity of the membrane's interior rather than a direct interaction with the valinomycin.     

Pyrethroid action on the nicotinic acetylcholine receptor/channel

The interactions of natural pyrethrins and nine pyrethroids with the nicotinic acetylcholine (ACh) receptor/channel complex of Torpedo electric organ membranes were studied. None caused significant reduction in [³H]ACh binding to the receptor sites, but all inhibited [³H]perhydrohistrionicotoxin ([³H]H₁₂-HTX) binding to the channel sites in presence of carbamylcholine. Allethrin inhibited [³H]H₁₂-HTX binding noncompetitively, but [³H]imipramine binding competitively, suggesting that allethrin binds to the receptor's channel sites that bind imipramine. The pyrethroids were divided into two types according to their actions: type I, which included pyrethrins, allethrin, bioallethrin, resmethrin, and tetramethrin, was more potent in inhibiting [³H]H₁₂-HTX binding and acted more rapidly (i.e., in <30 sec). Type II, which included permethrin, fluvalinate, cypermethrin and fenvalerate, was less potent and their potency increased slowly with time. Also, inhibition of the initial rate of [³H]H₁₂-HTX binding by type I compounds increased greatly by the presence of the agonist carbamylcholine, but this was not so with type II compounds. The receptor-regulated ⁴⁵Ca²⁺ flux into Torpedo microsacs was inhibited by pyrethrins and pyrethroids, suggesting that their action on this receptor function is inhibitory. There was very poor correlation between the potencies of pyrethrins and pyrethroids in inhibiting [³H]H₁₂-HTX binding and their toxicities to house flies, mosquitoes, and the American cockroach. However, the high affinities that several pyrethroids have for this nicotinic ACh receptor suggest that pyrethroids may have a synaptic site of action in addition to their well known effects on the axonal channels.     

Myotoxic and neurotoxic activity of Bacillus thuringiensis var. israelensis crystal toxin

The mechanism of the entomocidal action of Bacillus thuringiensis var. israelensis (BTI) in Periplaneta americana has been studied. Cockroaches treated with the alkali-solubilized BTI crystal gradually became sluggish and immobile. A physiological examination of poisoned cockroaches indicated that BTI possessed both myotoxic and neurotoxic activity. Following hemocoel or foregut administration of BTI, myotoxic effects were observed within 10–20 min whereas the onset of neurotoxic effects was considerably delayed. The results of this study suggest that the myotoxic action of BTI is probably important for the initial manifestation of its toxicity. The neurotoxic effects of BTI were ascribed to its ability to interfere with transmitter release. BTI exerted a dual action on transmitter release in the cockroach sixth abdominal ganglion. At lower doses (2–4 μg/ml) BTI was found to suppress transmitter release by interfering with calcium uptake. At higher concentrations (12 μg/ml or higher), however, BTI caused blockage of synaptic transmission which was preceded by massive transmitter release. In either case, the synapse blocking action of BTI was probably due to its effects upon presynaptic terminals. Postsynaptic membranes and axons in the ventral nerve cord apparently remained unaffected.     

Structure-activity relationships of DDT-type analogs based on in vivo toxicity to the sensory nerves of the cockroach, Periplaneta americana L

Forty-three DDT-type compounds were applied in saline suspension to the crural nerve of Periplaneta americana L. and the threshold concentration (ED₅₀) to produce trains of impulses was determined together with the frequency of appearance of repetitive afterdischarge. These quantitative neurological measures were evaluated in multiple regression analyses of structural parameters including van der Waal's volume, the F and R components of Hammet's σ, and the hydrophobic constant Π. This structure-activity analysis provides an accurate estimation of the intrinsic toxicity of the DDT analogs. The results affirm previous working theories that the bulk of the functional groups within the DDT framework is the primary factor relating to activity. However, conformation is also an important parameter.