Heterologous Expression of an Insecticidal Peptide Obtained from the Transcriptome of the Colombian Spider Phoneutria depilate.

Spider venoms are composed, among other substances, of peptide toxins whose selectivity for certain physiological targets has made them powerful tools for applications such as bioinsecticides, analgesics, antiarrhythmics, antibacterials, antifungals and antimalarials, among others. Bioinsecticides are an environmentally friendly alternative to conventional agrochemicals. In this paper, the primary structure of an insecticidal peptide was obtained from the venom gland transcriptome of the ctenid spider Phoneutria depilata (Transcript ID PhdNtxNav24). The peptide contains 53 amino acids, including 10 Cys residues that form 5 disulfide bonds. Using the amino acid sequence of such peptide, a synthetic gene was constructed de novo by overlapping PCRs and cloned into an expression vector. A recombinant peptide, named delta-ctenitoxin (rCtx-4), was obtained. It was expressed, folded, purified and validated using mass spectrometry (7994.61 Da). The insecticidal activity of rCtx-4 was demonstrated through intrathoracic injection in crickets (LD50 1.2 µg/g insect) and it was not toxic to mice. rCtx-4 is a potential bioinsecticide that could have a broad spectrum of applications in agriculture.

Study of the dermal anti-inflammatory, antioxidant, and analgesic activity of pinostrobin.

Pinostrobin is a flavanone isolated from Renealmia alpinia (Rottb.) Maas, which is used to treat painful diseases and ailments; indigenous peoples use it as plasters. Different plant species have been reported as a source of this flavonoid, among which are: Boesenbergia rotunda, Cajanus cajan, Piper ecuadorense, Piper hispidum, Teloxys graveolens, Kaempferia pandurata, among others. Pinostrobin expresses potentially useful biological activities such as antioxidant, analgesic, and dermal anti-inflammatory, at low levels nonetheless due to its low solubility. The formation of inclusion complexes deems a good strategy to improve the pharmacologic effects of many substances. In the present work, we evaluated the dermal toxicity, analgesic and dermal anti-inflammatory activity of pinostrobin included in cyclodextrins, to improve those effects on experimental animals. To include pinostrobin, we used two of beta cyclodextrin (ßCD) and hydroxypropil beta cyclodextrin (HPßCD) complexes using two methods developed by Benesi-Hildebrand and Higuchi-Connors. Dermal anti-inflammatory activity was evaluated in experimental mice by inhibiting the edema generated by 12-O-tetradecanoylforbol-13-acetate (TPA). Analgesic activity was evaluated by inducing chemical pain by means of a Siegmund test. Antioxidant activities were measured with two in vitro tests. Analgesic and dermal anti-inflammatory activities of pinostrobin, as included in control and experimental complexes, showed comparatively better effects than pinostrobin without inclusion complexes. Our results indicate that both beta cyclodextrin (ßCD) and hydroxypropyl beta cyclodextrin (HPßCD) enhance the different effects of pinostrobin, which may indicate greater bioavailability.