The systemic effects of juvenoids on the red firebug Pyrrhocoris apterus and on the pea aphid Acyrthosiphon pisum with data on life table response

BACKGROUND: A series of juvenoid alcohols and their glycosidic derivatives (juvenogens), synthesized at the Institute of Organic Chemistry and Biochemistry in Prague, commercially used juvenoids and the natural derivatives of juvabione were evaluated for their systemic juvenilizing effect on the red firebug, Pyrrhocoris apterus L., and on the pea aphid, Acyrthosiphon pisum Harris. A life table response experiment was designed, and demographic characteristics were computed for a cohort of A. pisum following chronic exposure to a range of concentrations of the selected trans-isomer of carbamate juvenoid 8 applied via the broad bean (Vicia faba L.) root system. RESULTS: Using the ratio of topical and per os activities, promising effects were found in two alkyl beta-D-glucopyranosides (3 and 9) and the trans-isomer of carbamate juvenoid 8 in tests on P. apterus. In A. pisum, the highest systemic activity was found in tests with the trans-isomer of carbamate juvenoid 8. The longevity, the number of offspring per female and the number of offspring per reproducing female significantly decreased with concentrations of 0.05 mg mL(-1) and higher. While the net reproductive rate R(0) and the intrinsic rate of natural increase r(m) displayed similar trends, the generation time G varied slightly between 11.6 and 12.8 days. CONCLUSION: Expected systemic activity of glycosidic juvenogens in P. apterus was not confirmed by exposure of the pea aphid on broad bean treated with aqueous solutions of the compounds. Nevertheless, the carbamate juvenoid alcohol 8 displayed a considerable juvenilizing effect on A. pisum in screening tests. Furthermore, this compound indicated a sublethal effect as the realized fecundity U(x) decreased disproportionately to the age-specific survival L(x) with increased concentration. The population parameters are compared with the data in similar demographic studies and are discussed together with the possibilities of using the compounds of this structural type in practice.     

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’).     

Insecticidal activity of N,N-diethyl-, N,N-dipropyl-, N,n-di-isopropyl-, and n,n-di-isobutylalkanamides against mosquito larvae

N,N-Diethyl-, N,N-dipropyl-, N,N-di-isopropyl, and N,N-di-isobutylalkanamides in which the acyl moiety ranged from C₈ to C₂₁ were synthesised, and their larvicidal activity was determined against the first-instar larvae of the southern house mosquito Culex quinquefasciatus Say. The four homologous series of amides generally showed an increase in their larvicidal activity as the carbon number in the acyl moiety of the amides increased, until the activity reached a maximum. Subsequently, an increment of carbon number resulted in declining activity in the higher homologues, until the activity disappeared. N,N-Diethyltetradecanamide, N,N-dipropylundecanamide, N,N-di-isopropylundecanamide, and N,N-di-isobutlynonanamide or -dodecanamide were the most active compounds in their respective homologous series of amides; however, they were less active than their analogous N,N-dimethylalkanamides previously studied.     

Juvenogens as potential agents in termite control: laboratory screening

Preliminary screening tests of eight juvenogens, fatty acid esters of two parent juvenoid alcohols, showed high juvenilizing effect in the termite Prorhinotermes simplex (Hagen). This effect is manifested by differentiation of termite workers into pre-soldiers, soldiers and/or soldier-intercastes ('soldiers'). Juvenogen 9 was tested with five other termite species: Zootermopsis angusticollis Hagen, Kalotermes flavicollis F, Cryptotermes declivis Tsai & Chen, Reticulitermes santonensis de Feytaud, and R flaviceps Oshima, and showed promising efficacy in force-feeding (FF) as well as in choice bioassays. In the economically important subterranean termites R santonensis and R flaviceps, after application of 0.5 mg ml(-1) in FF experiments ca 57 and 73%, respectively, of workers changed into soldiers. In Zootermopsis angusticollis application of 0.05 mg ml(-1) caused differentiation in 62% soldiers, and 57% soldiers differentiated after topical application of 1 microg of the tested compound per termite. Differentiation of excessive soldiers induced by a juvenile hormone-mimicking compound may cause disruption of the social structure and ultimately the death of the colony.     

Comparative inhibition of the juvenile hormone esterases from Trichoplusia ni,Tenebrio molitor, and Musca domestica

Twenty-seven compounds were screened as potential inhibitors of juvenile hormone esterases. Of these compounds O-ethyl-S-phenyl phosphoramidothiolate provided the best inhibition for the cabbage looper, Trichoplusia ni (Hubner), and the yellow mealworm, Tenebrio molitor L., while the juvenile hormone esterases of the house fly, Musca domestica L., were best inhibited by a juvenoid carbamate (1-(m-phenoxy-N-ethyl carbamate)-3,7-dimethyl-7-methoxy-2E-octene). The inhibition patterns of T. ni and T. molitor are similar, while those of M. domestica are relatively different. Further studies on the juvenile hormone and α-napthyl acetate esterases of T. ni showed that they could be differentially inhibited. Diisopropyl phosphorofluoridate and an alkyl trifluoromethyl ketone selectively inhibit the hydrolysis of α-naphthyl acetate and juvenile hormone, respectively, while O-ethyl-S-phenyl phosporamidothiolate inhibits both enzymes. The juvenile hormone esterases of T. ni also appear to be unique enzymes that are selective for juvenile-hormone-like molecules. The in vivo inhibition of T. ni juvenile hormone esterases by O-ethyl-S-phenyl phosphoramidothiolate slows the in vivo hydrolysis of juvenile hormone and results in delayed pupation and malformed larvae that resemble larval-pupal intermediates. Thus, the esterases involved in juvenile hormone metabolism appear to be important in juvenile hormone regulation.     

Synthesis and biological activity of furanyl anti-juvenile hormonal compounds

Twenty-one synthetic compounds, containing one or more furan rings, were demonstrated to possess anti-juvenile hormone (AJH) activity as evidenced by their induction of premature metamorphosis in the milkweed bug, Oncopeltus fasciatus (Dallas) by contact, topical application or fumigation. The ED₅₀ of the four most active analogs required to induce precocious metamorphosis from 3rd-instar nymphs by residue contact in a Petri dish compared favorably with that of precocene II (6,7-dimethoxy-2,2-dimethyl 2H-chromene) a naturally occurring phytochemical AJH. Precocious metamorphosis was fully reversible by co-treatment with juvenile hormone (JH III) or JH analogs, demonstrating that the observed AJH activity resulted from an induced deficiency of juvenile hormone.     

Effect of adjuvants on the performance of the new cereal fungicide,metconazole. I glasshouse trials

Metconazole, (1RS, 5RS; 1RS, 5SR)-5-(4-chlorobenzyl)-2,2-dimethyl-1-(1H-1,2,4-triazol-1-ylmethyl)cyclopentanol, is a highly active fungicide, in particular controlling seed-borne and foliar diseases of cereals, such as wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.). In order to maximize its foliar activity, an experimental survey of some types of surfactants and an emulsifiable oil was undertaken. Two types of metconazole formulation were investigated using a biological assay involving the therapeutic control of two diseases (Erysiphe graminis DC f. sp. tritici Marchal and Leptosphaeria nodorum Muell.) of wheat. Enhancements of activity by ?Genapol’? C12/C14 alcohol ethoxylates of an emulsifiable concentrate (ECM) formulation of metconazole were approximately three- to four-fold. For its, initially, less active suspension concentrate (SCM) formulation, enhancements were around 35-fold, so that with these adjuvants the two formulations were of similar activity. The enhancement ability of these ?Genapol’? surfactants was optimal at lower (5–10 moles) ethylene oxide content. These analogues induced marginally better enhancements of activity than members of a range of nonylphenol ethoxylates (?Arkopal’?), and emulsifiable paraffinic/naphthenic oil (HVI 60E) and a castor oil ethoxylate (?Atlas’? G1281) but were equivalent to a similar series of alcohol ethoxylates (?Dobanol’?) from another source. Varying the alkyl chain length between C₉-C₁₁ and C₁₄-C₁₅ in the ?Dobanol’? series had little effect on their high enhancements of metconazole activity. It was determined from trials varying the application rate of the best alcohol ethoxylates that application rates of 1–1.5 kg ha^?1 were required for maximum activity. This implied the use of high adjuvant/metconazole ratios in one-pack adjuvant-containing formulations. A range of soluble liquid (SL) formulations were prepared with either ?Dobanol’? 23–6.5 or ?Dobanol’? 91-6/metconazole ratios varying from 5:1 to 15:1. There were found to be highly active and were recommended for field testing.     

Effect of Adjuvants on the Therapeutic Activity of Dimethomorph in Controlling Vine Downy Mildew. I. Survey of Adjuvant Types

The effect of adjuvants on the performance of emulsifiable concentrate (EC) and wettable powder (WP) formulations of dimethomorph, a new systemic Oomycete fungicide, has been investigated using a two-day therapeutic (curative) assay with downy mildew (Plasmopara viticola, Berl. & de T.) on vines (Vitisvinifera L., cv. Cabernet Sauvignon) in glasshouse trials. The EC formulation had some therapeutic activity in this type of test. This activity was increased by the spray tank incorporation of 6 g litre⁻¹ of either emulsifiable rape seed oil (‘Atplus’ 412) or emulsifiable paraffinic oil (‘Atplus’ 411F). However, these improvements in performance were overshadowed by those brought about by incorporation of 3 g litre⁻¹ of a series of C₁₃/C₁₄ alcohol ethoxylates varying in ethylene oxide content from 5 to 20 moles:(‘Marlipal’ 34/6EO, 34/11EO, 34/20EO). Nearly complete fungal control was obtained in the presence of these adjuvants with a dimethomorph application rate of 25 g ha⁻¹ compared with only around 90% control at 400 g ha⁻¹ without adjuvants. The WP formulation was inactive in this therapeutic test but the presence of the adjuvants improved the performance of this formulation towards the high levels observed with the EC plus adjuvants, demonstrating that adjuvants could markedly influence the performance of solid, otherwise therapeutically inactive, dimethomorph formulations. Further trials examined other types of adjuvants (nonylphenol, alkylamine and silicone ethoxylates) but either they were no better than the alcohol ethoxylates or they induced unacceptable phytotoxicity. Trials with alcohol ethoxylates (‘Genapols’) from another source demonstrated activity equivalent to the ‘Marlipal’ surfactants. A two-factorial matrix experiment with ‘Genapol’ C050 showed that, under glasshouse conditions, >90% control could be obtained with the dimethomorph EC at 25 g AI ha⁻¹ with 375 g ha⁻¹ ‘Genapol’ C050. Applications of the WP formulation required slightly higher rates of either 50 g AI ha⁻¹ plus 375 g ha⁻¹ ‘Genapol’ C050 or 25 g AI ha⁻¹ plus 750–1500 g ha⁻¹ ‘Genapol’ C050. The overall conclusion was that alcohol ethoxylates varying in alkyl chain length from C₁₂ to C₁₈ and ethylene oxide content between 5 and 20 moles for the C₁₂ surfactants and ∽15 moles for the C₁₈ surfactants were effective adjuvants in promoting the therapeutic activity of dimethomorph formulationsagainst P. viticola on glasshouse-propagated vines.     

Biological and biochemical aspects of the disruption of adult formation in Tribolium by a novel bis(thiocarbamate) R-31026 and the juvenoid R-20458

S,S-Di-isobutyl N ethylethylenebis(thiocarbamate) (R-31026) had a strong morphogenetic effect, acting by contact on the newly formed pupae of Tribolium confusum and Tribolium castaneum and thereby disrupting adult development. At a dietary concentration of 20 mg kg^?1, 95% of pupae of T. confusum produced pupa-adult intermediates; T. castaneum pupae were affected to a smaller extent. Topical application with 0.002 μg per 0–24-h-old pupa of T. confusum resulted in the formation of 88% malformed intermediates. The larvae brought into contact with R-31026 were not affected and pupated normally, hence the compound differs in its activity from that of a typical juvenile hormone compound. On the other hand, the pupal morphogenetic activity of R-31026 resembles that of the typical juvenoid compound 3-[5-(4-ethylphenoxy)-3-methylpent-3-enyl]-2,2-dimethyloxirane (R-20458). The progeny of the emerging adults from pupae treated with either R-20458 or R-31026 were strongly affected. The effective dosages were far below those required for pupal morphogenetic activity. Biochemical studies showed an increase in the soluble protein fraction during the pupal stage after treatment with either R-31026 or R-20458 indicating disturbances in protein build-up. The bis(thiocarbamate) R-31026 has more favourable practical properties than R-20458 for controlling agricultural insects, because it does not prolong the larval feeding stage.     

The action of pyrethroids in the insect central nervous system. I. Features of molecular structure associated with toxicity to cockroaches and to their giant fibre axons

To investigate relationships between the molecular structure of pyrethroids and their mode of action, toxicities to adult male Periplaneta americana by topical application and injection were compared with toxicities to their giant fibre nerve axons. From the tests against intact insects it was concluded that: (i) although ED₅₀S ranged from 0.04 to 65 μg/insect, each compound was equally toxic, with one exception, when administered by either route; (ii) esters of (1 R)-cis- were more toxic than esters of the corresponding (1 R)-trans-3-substituted-2, 2-dimethylcyclopropanecarboxylic acids; (iii) α-cyano-3-phenoxybenzyl esters were more toxic than the corresponding 3-phenoxybenzyl esters; (iv) changes in the alcoholic component of some compounds (particularly trans-isomers of esters of 5-benzyl-3-furylmethanol and esters of α-cyano-3-phenoxybenzyl alcohol) affected a recovery phase in their ED₅₀/time curves more than changes in the acid component; (v) the amount of recovery was positively correlated with molecular polarity. The concentration required to decrease the amplitude of the action potential of giant fibres by 30% in 1 h ranged from 0.26 μM for the most active compound to 100 μM for the least active. There was no clear relationship between neurotoxicity and toxicity to whole insects and little association between neurotoxicity and features of molecular structure. Neurotoxicity was, however, positively correlated with polarity. Giant fibre axons seem unlikely to be critical sites of action of pyrethroids.     

Selective inhibition of JH esterases from cockroach hemolymph

Juvenile hormone III was tritium labeled on the methyl ester and utilized with other substrates in an investigation of inhibition and substrate specificity of hemolymph esterases from the cockroach, Blaberus giganteus. The structure of labeled juvenile hormone III was supported both chemically and biochemically. Forty-two potential inhibitors were examined, and the best inhibitors included phosphoramidothiolates and S-phenylphosphates. One of these inhibitors was found useful in hormone biosynthesis studies dealing with the enzymatic conversion of methyl farnesoate to juvenile hormone in corpora allata homogenates. Several commonly used inhibitors of carboxyesterases caused only weak inhibition of JH esterases. Gel filtration elution patterns, inhibitor relationships, and specific activities of the hemolymph esterases indicate that juvenile hormones I and III are degraded by similar if not identical enzymes. In some cases, α-naphthyl acetate and juvenile hormone esterase activity could be differentially inhibited. Hemolymph esterases were not capable of degrading ethyl or isopropyl conjugated esters of two juvenoids or three model substrates.     

Juvenile hormone III, hydroprene and a juvenogen as soldier caste differentiation regulators in three Reticulitermes species: potential of juvenile hormone analogues in termite control

The efficacy of juvenile hormone III (JH III) and two JH-mimicking compounds was compared in laboratory experiments with Reticulitermes lucifugus (Rossi), R. santonensis Feytaud and R. virginicus (Banks). Induction of presoldier, soldier and/or soldier-worker intercaste differentiation was taken as positive response to the treatment. The novel juvenogen, a fatty acid ester of a juvenoid alcohol, induced greatest soldier differentiation in all species tested, followed by hydroprene. JH III was less effective. Representatives of three Reticulitermes species showed similar trends in soldier induction rates. Differences in mortality in treatments of termites of the same species from different colonies with the same compound were observed and evidently were caused by differences in the conditions of respective colonies.     

Substituted thiotrifluoropropanones as potent selective inhibitors of juvenile hormone esterase

A series of 27 substituted thio-1,1,1-trifluoropropanones was synthesized by reacting the corresponding thiol with 1,1,1-trifluoro-3-bromopropanone. The resulting sulfides were screened as inhibitors of hemolymph juvenile hormone esterase and α-naphthyl acetate esterase activity of the cabbage looper, Trichoplusia ni, electric eel acetylcholinesterase, bovine trypsin, and bovine α-chymotrypsin. The presence of the sulfide bond increased the inhibitory potency on all of the enzymes tested when compared with compounds lacking the sulfide. In general, the compounds proved to be poor inhibitors of chymotrypsin and moderate inhibitors of trypsin. By varying the substituent on the sulfide, good inhibitory activity was obtained on α-naphthyl acetate esterase, acetylcholinesterase, while some of the compounds proved to be extremely powerful inhibitors of juvenile hormone esterase. The most powerful inhibitor tested was 3-octylthio-1,1,1-trifluoro-2-propanone, with an I₅₀ of 2.3 × 10^?9M on JH esterase. This compound showed a molar refractivity similar to that of the JH II backbone, was not toxic to T. ni, and was moderately toxic to mice, with a 48-hr LD₅₀ of >750 mg/kg. It effectively delayed pupation when applied to prewandering larvae of T. ni, as expected for a JH esterase inhibitor. Thus, some members of this series are promising for evaluating the role of JH esterase in insect development. The series also indicates that, by varying the substituent on the sulfide moiety, potent “transition-state” inhibitors can be developed for a wide variety of esterases and proteases.     

Inhibition of epoxidation of methyl farnesoate to juvenile hormone III by cockroach corpus allatum homogenates

Radiolabeled methyl farnesoate is epoxidized to juvenile hormone III by an NADPH-dependent reaction occurring in corpus allatum homogenates from the cockroach Blaberus giganteus L. Most of the enzymatically produced juvenile hormone has the 10R configuration described for previously isolated natural juvenile hormones. The unnatural 2Z geometrical isomer of methyl farnesoate is epoxidized by the above system faster than the natural 2E isomer. Several series of chemicals known to be inhibitors of mixed-function oxidases were surveyed as inhibitors of methyl farnesoate epoxidation. The anti-juvenile hormone precocene II caused negligible inhibition at 1 · 10^?4M, whereas the best inhibitor was o-bromophenoxymethyl-imidazole with an apparent I₅₀ of 4 · 10^?7M. None of the inhibitors tested were potent morphogenetic agents on Tenebrio molitor pupae, and they failed to cause precocious development of Oncopeltus fasciatus nymphs. The inhibition of in vitro juvenile hormone biosynthesis suggests the possibility of finding an anti-hormone which acts by blocking juvenile hormone biosynthesis.     

Structure—activity relationships in pyrethroid esters derived from substituted 2-phenoxy-3-methylbutanoic acids

Non-cyclopropane pyrethroid esters of different substituted 2-phenoxy-3-methylbutanoic acids have been synthesised using the three alcohols—3-phenoxybenzyl alcohol, α-cyano-3-phenoxybenzyl alcohol and 3, 4-methylene-dioxybenzyl alcohol. Among the 35 esters synthesised and tested against Culex quinquefasciatus Say, the Bancroftian filariasis vector, for both larvicidal and adulticidal activities, α-cyano-3-phenoxybenzyl 2-(4-fluorophenoxy)-3-methylbu-tanoate, with an LC₅₀ value of 2.5 × 10^?3 mg litre^?1 for larvicidal activity, and α-cyano-3-phenoxybenzyl-2-(4-chlorophenoxy)-3-methylbutanoate, with an LD₅₀ value of 30 times; 10^?4 ug insect^?1 for adulticidal activity, were found to be as effective as fenvalerate, a well-known non-cyclopropane pyrethroid ester. Structure-activity studies showed that the insecticidal activity is dependent on the nature and position of the substituent in the phenyl ring of the acid moiety and also on the type of alcohol moiety.     

The inhibition of insect juvenile hormone esterase by trifluoromethylketones: Steric parameters at the active site

A rationally designed structure-activity relationship study has been accomplished using trifluoromethylketone inhibitors of insect juvenile hormone esterase from the cabbage looper, Trichoplusia ni (Hubner) (Lepidoptera: Noctuidae). Several α- and α′-substituted derivatives of 3-octylthio-1,1,1-trifluoropropan-2-one have been prepared and assayed for inhibitory potency against juvenile hormone esterase. The results indicate that the sulfur/protein interaction does not occur in a sterically constrained environment. Substitution adjacent to sulfur did not dramatically effect activity. However, substitution adjacent to the carbonyl of the trifluoromethylketone moiety reduced inhibitory potency substantially, indicating that the active site region of juvenile hormone esterase which interacts with the carbonyl is restricted to rather small substrates. A small hydrophobic pocket near the active site has been identified and can serve to increase inhibitory potency by secondary binding of appropriate substituents. The present study has resulted in the preparation of two more effective in vitro inhibitors of juvenile hormone esterase than those previously reported. Evidence that there are two naturally occurring forms of juvenile hormone esterase has also been provided.     

Inhibition of juvenile hormone biosynthesis in the tobacco hornworm, Manduca sexta, by a bisthiolcarbamate juvenoid and related compounds

Biosynthesis of juvenile hormone in the tobacco hornworm, Manduca sexta, is inhibited by the bisthiolcarbamate juvenoid N-ethyl-1,2-bis(isobutylthiolcarbamoyl)ethane both in vitro and in vivo. In vitro an extremely steep dose-response curve was obtained with an ID₅₀ value of 6 × 10^?6M. However, in vivo topical treatment with the compound resulted in mild JH antagonistic symptoms, suggesting rapid metabolism of the compound. In agreement with results from metabolic studies performed on plants and in mammals, sulfoxidation of the thiocarbamate S-(4-chlorobenzyl)N,N-diethylthiocarbamate resulted in an enhanced inhibitory effect on JH biosynthesis in vitro. This suggests that the corresponding thiocarbamate sulfoxides may act as intermediates in carbomylating critical thiol sites important in the terpenoid biosynthesis pathway. Furthermore, this study shows that these prototype compounds are interesting tools for further investigation of chemical inhibition of JH biosynthesis in insects.     

Molecular modifications of benzylphenol and benzyl-1,3-benzodioxole types of insect chemosterilants

Rationally planned structural modifications were carried out on benzylphenols and benzyl-1,3-benzodioxoles described as fly chemosterilants and as anti-juvenile hormones. The introduction of a prop-2-ynyloxy group at various sites of the molecule resulted in compounds with a moderate inhibitory action on cytochrome P-450 mono-oxygenases, as measured by aldrin epoxidation. One compound, 5-(4-methoxybenzyl)-6-prop-2-ynyloxy-1,3-benzodioxole, revealed chemosterilant activity on Phormia regina, but its activity was less than that of the parent compounds. 2,4-Di-tert-butyl-6-[4-(3-methoxy-3-methylbutoxy)benzyl]phenol, which possessed a juvenoid structure, revealed no juvenile hormone (JH) activity but showed a high sterilant effect against Dysdercus cingulatus. In contrast to the parent substances, none of the tested compounds showed a detectable anti-JH effect in the Galleria assay. 8-Methoxy-2,3-methylenedioxydibenz[b,e]oxepine, a hitherto undescribed fused heterocyclic ring system, was devoid of activity, indicating the importance of free rotation and/or molecular flexibility. In spite of the moderate activities of these compounds, the manifold biological potential of the quinone-methide mechanism justifies further research on these lines.     

Albumin and cytochrome P450 binding characteristics of juvenile hormone and its analogs

Binding data were gathered for the cecropia juvenile hormone (methyl(E, E cis)-10,11-epoxy-7-ethyl-3,11-dimethyl-2,6-tridecadienoate) and two of its analogs {isopropyl(2E, 4E)-11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate; (E)-4-[(6,7-epoxy-3,7-dimethyl-2-nonenyl)-oxyl]-1,2-(methylenedioxy)benzene} with bovine serum albumin and rat hepatic microsomal cytochrome P₄₅₀. The proteins were found to bind the juvenile hormone and juvenile hormone analogs with affinity constants ranging from 10⁵ to 10⁶M^?1. Thermodynamic calculations suggest that the binding of all three compounds is electrostatic in nature and that the size of the ether and ester substituents can greatly influence the binding to proteins. The juvenile hormone and its analogs all formed spectrally apparent Type I complexes with oxidized cytochrome P₄₅₀; one of the juvenile hormone analogs formed a spectrally observable product adduct with reduced cytochrome P₄₅₀. The product complex may contribute many of the hormonal effects observed for this compound.