Growing resistance to insecticides, especially pyrethroids, poses an increased risk for insect control world-wide. Thus, alternative compounds are urgently needed. Accordingly, my laboratory has been investigating synthetic materials, as well as natural products, which are an attractive source of new chemistries. We have examined the insecticidal and synergistic activity of new compounds using Drosophila melanogaster and the mosquitoes Aedes aegypti and Anopheles gambiae as model insects. In addition, we have also been investigating the modes of action of experimental molecules in various physiological preparations. Natural product research included thymol, which was toxic to Ae. aegypti adults at doses (3-50 μg/mg) similar to published toxicities for the cabbage looper larvae, Trichoplusia ni. On homomultimeric Anopheles gambiae GABA receptor-chloride channel complex expressed in Xenopus laevis oocytes, thymol acts as a positive allosteric modulator, increasing the potency and maximal effectiveness of applied GABA, consistent with the sluggish paralysis it elicits in exposed insects. Other studies investigated matrine; a bioactive component extracted from Sophora flavescens that is used as the main ingredient in Chinese bio-pesticidal products. Matrine caused flaccid paralysis in headless fourth instar larvae of Ae. aegypti (50% paralysis in 5 hours at 8 ppm) and was toxic to adult females by contact (topical LD50 = 258 ng/mg). Adult toxicity was increased about 2-fold by pretreatment with the mono-oxygenase inhibitor, piperonyl butoxide. Interestingly, this compound was much less effective on D. melanogaster in either glass contact or feeding bioassays. Adult mosquito knockdown was rapid, with little or no expression of hyperactivity or hyperexcitability. Matrine (ca. 1 mM) had effects similar to thymol on expressed An. gambiae GABA receptors, and reduced EPSP amplitude at the Musca domestica neuromuscular junction, without any evidence of neuroexcitation or membrane depolarization. These physiological actions are sufficient to explain the whole animal intoxication by matrine, but require relatively high concentrations to manifest themselves. Other studies tested a series of potassium channel blockers, including 1-((2-chlorophenyl)diphenylmethyl)-1H-pyrazole (TRAM-34), 11-dansylaminoundecanoic acid (DAUDA), and 5-hydroxydecanoic acid (5-HDC) as insecticides and synergists of pyrethroids. We also evaluated the action of these compounds in patch clamp recordings of engineered HEK cells expressing Anopheles gambiae Kv2.1 channels. Patch clamp studies revealed that fatty acid compounds without functional groups in the alkyl chain (e.g., decanoate, DAUDA) yielded a more potent blocking action on Kv2.1 currents than substituted fatty acids (e.g., 5-HDC). Also, in comparison to 2-methoxy-N-((1-phenylcyclopentyl)methyl)benzamide (2S-65465), a known Kv2 channel blocker (IC50 = 100 nM), decanoate and DAUDA were 6-fold and 12-fold less active, respectively, as blockers of potassium current. TRAM-34 was the least potent inhibitor tested in patch clamp studies (IC50 = 30 uM). When tested on D. melanogaster CNS, the compounds typically gave an initial increase in firing rate, followed by a decrease, both effects at micromolar concentrations. Various toxicity assays showed the same potency ranking as that obtained through patch-clamp recordings, indicating a possible connection between channel block and whole organism effects. In order to possibly improve the insecticidal activity of the fatty acids, we synthesized derivatized analogs (e.g., methyl esters) presumed to better penetrate the cuticular barrier, but activity did not improve. Finally, in contrast to a published patent, we found that fatty acids did not show much synergism with pyrethroids. The implications of this work for resistance management will be discussed.