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.

몇가지 협력제의 7-ethoxy precocene II와 협력작용을 통한 활성증가와 작용기작을 구명하기 위하여 mikweed bug, Oncopeltus fasciatus 2령충에 항유약호르몬유사물과 협력제가 처리된 petri dish에 접종하여 생리활성 정도를 측정하였다. 얻어진 결과는 다음과 같다. 7-Ehtoxy precocene II의 반수활성농도는 g/이었다. 7-Ethoxy precocene II와 다른 산화저해제인 RO20-9747의 협력비가 14.05로 가장 높았고 그들의 반수활성농도는 이었다. 7-Ethoxy precocene II와 다른 협력제 1종, 2종, 4종을 조합을 달리하여 처리하였을 때 뚜렷한 협력작용 증가효과를 가져오지 못했다. 이상의 결과에서 다른 협력제들은 곤충의 방어기작을 알라타체를 포함하여 비선택적으로 저해하는 것으로 생각되었으나 RO20-9747은 7-ethoxy precocene II가 알라타체로 이행하는데 까지 곤충의 방어적 산화기작인 monooxygenase 활성을 선택적으로 저해함으로써 7-ethoxy precocene II가 알라타체로 이행량이 증가하여 활성증가를 가져온 것으로 생각되었다.

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious clinical and an urgent problem worldwide. Few new drugs are available against MRSA, because MRSA has the ability to acquire resistance to most antibiotics, which consequently increases the cost of medication. In the present study, the antibacterial activity of Hydnocarpi Semen was investigated. The most effective method is to develop antibiotics from the natural products without having any toxic or side effects. Therefore, there is a need to develop alternative antibacterial drugs for the treatment of infectious diseases. Five Clinical isolates (MRSA) were obtained from five different patients at Wonkwang University Hospital (Iksan, South Korea). The Other 2 strains were ATCC 33591 (Methicillin-resistant strain) and ATCC 25923 (Methicillin-susceptible strain). Antibacterial activity (Minimal Inhibitory Concentrations, MICs) was determined by broth dilution method, disk diffusion method, MTT test, and checkerboard dilution test. Antibacterial activity of n-hexane fraction was remarkable, and had a MICs ranging from 31.25-125 ㎍/㎖. FICI values for HFH+AM and HFH+OX were 0.13-0.19 and 0.04-0.29, showing the increase of synergistic effect. When combined together, these antibacterial effects were dramatically increased.