Nisha Mathew

Antifilarial Lead Molecules Isolated from Trachyspermum ammi

Lymphatic filariasis is caused by infection with the parasitic filarial nematodes Wuchereria bancrofti, Brugia malayi and B. timori, transmitted by mosquitoes. The lack of an adulticidal drug poses a challenge to filariasis elimination, hence it is essential to develop an effective antifilarial drug which could either kill or permanently sterilize the adult worms. In the reported work the in vitro activity of a methanolic extract of fruits of Trachyspermum ammi (Apiaceae) against adult bovine filarial Setaria digitata worms has been investigated. A bioassay-guided fractionation was carried out by subjecting the crude extract to flash chromatography. HPLC analysis was done for the crude extract and active fraction. The crude extract and the active fraction showed significant activity against the adult S. digitata by both a worm motility and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] reduction assays. The isolated active principle was chemically characterized by IR, 1H-NMR and MS analysis and identified as a phenolic monoterpene. It was screened for in vivo antifilarial activity against the human filarial worm B. malayi in Mastomys coucha, showing macrofilaricidal activity and female worm sterility in vivo against B. malayi. The findings thus provide a new lead for development of a macrofilaricidal drug from natural products.

N,N-Diethyl Phenylacetamide (DEPA): A Safe and Effective Repellent for Personal Protection Against Hematophagous Arthropods

Abstract Repellents play an important role in protecting humans from the bites of insect pests. An effective and safe repellent will be useful in reducing human-vector contact and thereby help in the interruption of vector borne disease transmission. Because of the unavailability of m-toluic acid in India for the manufacture of N,N-diethyl m-toluamide (DEET), there is a need to develop an alternate effective and safe insect repellent. In total, 120 substituted amides were synthesized and tested for repellency at 1.0 mg/cm2 under laboratory conditions. Among these amides, N,N-diethyl phenylacetamide (DEPA), applied at 1.0 mg/cm2 in different oil bases, was found to exhibit promising repellency (6–8 h) in the laboratory when tested against Aedes aegypti (L.) The repellent DEPA was evaluated on army personnel in comparison with dimethylphthalate (DMP) and DEET against mosquitoes, black flies, and land leeches under field condition in the North-East Frontier area of India. Both DEPA and DEET displayed broad-spectrum repellency. DEPA was more effective than DMP against all test organisms. However, no significant difference was noticed between DEPA and DEET for repellency at 0.25 and 0.5 mg/cm2 against black flies and mosquitoes. DMP was the least effective among the three compounds in the field studies. The relative potency of DEPA in comparison with DEET and DMP for repellency against Phlebotomine sand flies also was determined. At 0.1 mg/cm2, both DEPA and DEET were found to be equally effective with a protection time from 4.37 ± 0.08 to 4.45 ± 0.15 h. Both compounds were significantly more effective than DMP. At 0.2 mg/cm2, DEPA and DEET provided protection times of 6.52 ± 0.08 and 7.15 ± 0.15 h, respectively. DEPA was formulated into a vanishing cream, a pharmacologically safe polymer-based liquid, and a liposphere lotion. The vanishing cream and the two-polymer liquid formulations enhanced protection times from 4.4 to 6.5 and 7.13 h, respectively, compared with an alcohol solution applied at 0.5 mg/cm2 on the forearm of human volunteers. Formulations of DEPA and DEET applied at 0.5 mg/cm2 on rabbits exposed to Ae. aegypti enhanced protection times compared with an alcohol solution and the liposphere lotion from 4.0 to 6.0 h and 4.0 to 5.0 h, respectively. Comparison of mean protection times of DEET and DEPA applied at a rate of 0.3 mg/cm2 to human volunteers exposed to Cx. quinquefasciatus under field conditions indicated that the lotion formulations of DEET and DEPA provided significantly (Duncan’s multiple-range test, P < 0.05) higher protection times than did alcohol solutions. Toxicological studies revealed that DEPA is safe.

Glutathione S-transferase (GST) inhibitors

The glutathione S-transferases (GSTs; EC 2.5.1.18) comprise a family of widely distributed Phase II detoxication enzymes that catalyse the conjugation of a broad variety of reactive electrophiles to the nucleophilic sulfur atom of the major intracellular thiol, the tripeptide glutathione. The diverse functions, including catalytic GSH conjugation, passive ligandin-type binding and modulation of signal transduction, may be selectively targeted by different inhibitors. GST inhibitors are emerging as promising therapeautic agents for managing the development of resistance amongst anticancer agents. In diagnostic medicine, as well as in antiparasitic drug development, GST inhibitors are important lead molecules. In this review, the important molecules known for their GST inhibition together with potential therapeutic uses are summarised.

Mosquito attractant blends to trap host seeking Aedes aegypti

Aedes aegypti is the key vector of three important arboviral diseases -dengue, yellow fever and chikungunya. To identify volatile chemicals which could be used in odour based traps for Aedes mosquito surveillance, a few synthetic compounds and compound blends have been evaluated in an indigenously designed olfactometer. A total of 24 compounds and seven compound blends were screened against unfed adult female Ae. aegypti mosquitoes for attraction and compared with control group. The attractancy or repellency index of the test material to mosquitoes was calculated and rated them as class-1, class-2 and class-3 with rating values ranging 1–15, 16–33 and 34–100 respectively. Out of the 24 compounds tested, six were showing significant attractancy (P < 0.05) and among that 1-octene-3-ol showed maximum attractancy with a rating value of 57.81. Sixteen compounds showed significant repellency (P < 0.05) and among that with a rating value of 72.47, 1-hexene-3-ol showed strong repellent action against Ae. aegypti. All the seven blends showed significant mosquito attractancy (P < 0.05) and among that with a rating of 62.08 Myristic acid, Lactic acid and CO2 blend exhibited first-rate mosquito attractancy.

A high performance liquid chromatographic method for the estimation of diethylcarbamazine content in medicated salt samples

A simple and reproducible method for the estimation of diethylcarbamazine citrate (DEC) by high performance liquid chromatography (HPLC) in DEC-medicated salt was developed. HPLC analysis was conducted with a mobile phase containing acetonitrile/phosphate buffer (20 mM KH(2)PO(4,) adjusted to pH 3.2 with 10% ortho-phosphoric acid) in the ratio of 1:9 and at a flow rate of 1.5 ml/min. A Phenomenex C8 column (15 cmx4.6 mm) of 5 microm particle size was used for the analysis. Analysis was done at UV 210 nm, 0.02 a.u.f. and 40 degrees C. The coefficient of variation was <10% in the range of 1-25 microg/ml and the minimum detectable level was 0.5 microg/ml. The quality of DEC-medicated salt prepared by two methods was analyzed by using the HPLC method. In spray drying method, 29 and 71% of the samples and in rotating drum method, 9 and 12% of samples were found to contain DEC at 0.15-0.25% and >0.25%, respectively. Thus, this quick and simple HPLC method for the estimation of DEC could play a vital role in checking the quality of the DEC medicated salt used for the control of filariasis.

Synthesis and screening of 1-methyl-4-substituted benzoyl piperazides against adult Setaria digitata for antifilarial activity

There is no safe and effective drug for killing the adult worms for the elimination/control of lymphatic filariasis and research is very much warranted towards the development of a macrofilaricidal drug. Therefore, the synthesis and evaluation of 1-N-methyl-substituted benzoyl/phenyl acetyl piperazides for macrofilaricidal activity were carried out. The title compounds, 1-N-methyl-substituted benzoyl/phenyl acetyl piperazides were synthesized and purified by slightly modifying the reported procedure and characterized by FT-IR, NMR and mass spectral data. The compounds were screened for macrofilaricidal activity against adult Setaria digitata, the bovine filarial worm for motility and MTT reduction assays using the reported procedures. All the compounds synthesized were characterized by spectral data. Out of 26 compounds synthesized and screened for macrofilaricidal activity, six compounds exhibited moderate antifilarial activity. The structure-activity relationships are discussed. In the case of benzoyl piperazides chloro-substitution in the para position enhanced the activity compared to its substitution in meta and ortho positions. In the case of phenyl acetyl piperazides when methyl group was in the meta position it was more active than when it was in para position. In dimethyl substituted compounds, substitutions at 3, 5-positions were more effective than 2, 3 and 3, 4 substitutions.

Development of a mosquito attractant blend of small molecules against host-seeking Aedes aegypti.

A mosquito’s dependence on olfaction in the hunt for human host could be efficiently exploited to protect humans from mosquito bites. The present study is undertaken to make the most attractant compound blend for Aedes aegypti mosquitoes to lure them to traps. Eleven molecules (M1–M11) at different dilutions were screened for attractancy against non-blood-fed adult female mosquitoes in an olfactometer. The results showed that the attractancy was dependent on both the chemical nature of the molecule and the strength of the odor. Out of 11 molecules screened, 9 showed significant attractancy (P < 0.05) when tested individually. The attractancy was in the order of M11 > M7 > M6 > M10 > M9 > M3 > M2 > M1 > M4 with attractancy indices (AIs) 86.11, 55.93, 55.17, 54, 52.94, 52, 50, 43.64, and 32, respectively, at the optimum dilutions. Seven blends (I–VII) were made and were screened for attractancy against Ae. aegypti. All the blends showed significant attractancy (P < 0.05). The attractancy was in the order of blend VII > III > IV > I > VI > V > II with AIs 96.63, 89.19, 65, 57.89, 56.1, 47.13, and 44.44, respectively. Among the seven blends, blend VII with constituent molecules M6, M9, M10, and M11 is the most promising with an AI value of 96.63. This blend will be useful in luring the host-seeking mosquitoes to traps. The field efficacy of these attractant blends may be explored in the future.

Synthesis and Macrofilaricidal Activity of Substituted 2-Hydroxy/5-Hydroxy/2-Methyl-1,4-Naphthoquinones

Preclinical Research