Rationally Designed α-Conotoxin Analogues Maintained Analgesia Activity and Weakened Side Effects.

A lack of specificity is restricting the further application of conotoxin from Conus bullatus (BuIA). In this study, an analogue library of BuIA was established and virtual screening was used, which identified high α7 nicotinic acetylcholine receptor (nAChR)-selectivity analogues. The analogues were synthesized and tested for their affinity to functional human α7 nAChR and for the regulation of intracellular calcium ion capacity in neurons. Immunofluorescence, flow cytometry, and patch clamp results showed that the analogues maintained their capacity for calcium regulation. The results of the hot-plate model and paclitaxel-induced peripheral neuropathy model indicated that, when compared with natural BuIA, the analgesia activities of the analogues in different models were maintained. To analyze the adverse effects and toxicity of BuIA and its analogues, the tail suspension test, forced swimming test, and open field test were used. The results showed that the safety and toxicity of the analogues were significantly better than BuIA. The analogues of BuIA with an appropriate and rational mutation showed high selectivity and maintained the regulation of Ca2+ capacity in neurons and activities of analgesia, whereas the analogues demonstrated that the adverse effects of natural α-conotoxins could be reduced.

Conotoxin MVIIA improves cell viability and antioxidant system after spinal cord injury in rats.

This study evaluates whether intrathecal MVIIA injection after spinal cord injury (SCI) elicits neuroprotective effects. The test rats were randomly distributed into six groups- sham, placebo, MVIIA 2.5 µM, MVIIA 5 µM, MVIIA 10 µM, and MVIIA 20 µM-and were administered the treatment four hours after SCI. After the optimal MVIIA dose (MVIIA 10 µM) was defined, the best time for application, one or four hours, was analyzed. Locomotor hind limb function and side effects were assessed. Forty-eight hours after the injury and immediately after euthanasia, spinal cord segments were removed from the test rats. Cell viability, reactive oxygen species, lipid peroxidation, and glutamate release were investigated. To examine the MVIIA mechanism of action, the gene expressions of pro-apoptotic (Bax, nNOS, and caspase-3, -8, -9, -12) and anti-apoptotic (Bcl-xl) factors in the spinal cord tissue samples were determined by real-time PCR, and the activities of antioxidant enzymes were also investigated. Application of intrathecal MVIIA 10 µM four hours after SCI prompted a neuroprotective effect: neuronal death decreased (22.46%), oxidative stress diminished, pro-apoptotic factors (Bax, nNOS, and caspase-3, -8) were expressed to a lesser extent, and mitochondrial viability as well as anti-apoptotic factor (Bcl-xl) expression increased. These results suggested that MVIIA provided neuroprotection through antioxidant effects. Indeed, superoxide dismutase (188.41%), and glutathione peroxidase (199.96%), reductase (193.86%), and transferase (175.93%) expressions increased. Therefore, intrathecal MVIIA (MVIIA 10 µM, 4 h) application has neuroprotective potential, and the possible mechanisms are related to antioxidant agent modulation and to intrinsic and extrinsic apoptotic pathways.

Human injuries and fatalities due to venomous marine snails of the family Conidae.

OBJECTIVE: This paper provides the first compilation in more than 30 years of human injuries and fatalities from envenomation by marine gastropod molluscs of the predominantly tropical family Conidae. It seeks to apply recent advances in knowledge of the physiological effects of conopeptides and molecular genetics to improve our understanding of the human responses to stings by species that normally use their venom peptides to paralyze and overcome prey such as polychaete worms, other gastropod molluscs, and fishes. RESULTS: A database has been constructed for the 139 cases accepted as reliably reporting each human injury. It includes data on the species responsible, the time and place where the stinging occurred and the sting site on the victim's body, the time-course of clinical effects, treatment carried out, if any, and outcome. Members of the hyperdiverse genus Conus caused all the injuries, except for 2 cases involving species from the recently separated genus Conasprella. Death occurred in 36 cases, 57 cases presented with serious symptoms but recovered completely, and in 44 cases victims were only minimally affected. A few cases are listed as tentative because the information in the reports was limited or unverifiable. Many cases have undoubtedly gone unreported and been forgotten. No cases are known for the period between the date of the first reliable report in the 17th century, and the mid-19th century. Knowledge of conopeptide molecular structure and function has recently burgeoned, permitting initial exploration of relationships between the symptoms and outcomes of human injuries and modes of action of these mainly small, very toxic neuroactive peptides. These relationships are reviewed here, especially in regard to the severe and fatal cases, with the aim of making recent knowledge accessible to clinicians and others involved in treating the effects of human stings, which continue to be reported. CONCLUSIONS: Conus geographus, a specialized predator of fishes, which it paralyzes with its venom and swallows whole, is the most dangerous species to humans. It accounts for about half of the known human envenomations and almost all the fatalities. Children succumb more often to C. geographus stings than adults and stings by larger snails are lethal more often than stings from smaller snails, regardless of the victim's age. Other piscivorous Conus species have stung humans, but with nonlethal results. A few species that normally prey on other gastropods have also seriously injured humans, but most of the fatalities reported have not been confirmed. Most species of Conidae prey only on marine worms; 18 of these species are known to have stung humans, with generally mild effects. Research on the treatment of Conus stings has lagged behind that on the application of conopeptides in pharmacological research and in the development of new pharmaceuticals. However, improved communication and availability of medical aid in remote tropical areas has likely contributed to reducing the mortality rate during the last half century.

Conotoxin Interactions with α9α10-nAChRs: Is the α9α10-Nicotinic Acetylcholine Receptor an Important Therapeutic Target for Pain Management?

The α9α10-nicotinic acetylcholine receptor (nAChR) has been implicated in pain and has been proposed to be a novel target for analgesics. However, the evidence to support the involvement of the α9α10-nAChR in pain is conflicted. This receptor was first implicated in pain with the characterisation of conotoxin Vc1.1, which is highly selective for α9α10-nAChRs and is an efficacious analgesic in chronic pain models with restorative capacities and no reported side effects. Numerous other analgesic conotoxin and non-conotoxin molecules have been subsequently characterised that also inhibit α9α10-nAChRs. However, there is evidence that α9α10-nAChR inhibition is neither necessary nor sufficient for analgesia. α9α10-nAChR-inhibiting analogues of Vc1.1 have no analgesic effects. Genetically-modified α9-nAChR knockout mice have a phenotype that is markedly different from the analgesic profile of Vc1.1 and similar conotoxins, suggesting that the conotoxin effects are largely independent of α9α10-nAChRs. Furthermore, an alternative mechanism of analgesia by Vc1.1 and other similar conotoxins involving non-canonical coupling of GABAB receptors to voltage-gated calcium channels is known. Additional incongruities regarding α9α10-nAChRs in analgesia are discussed. A more comprehensive characterisation of the role of α9α10-nAChRs in pain is crucial for understanding the analgesic action of conotoxins and for improved drug design.

Cutaneous abscess after Conus textile sting.

We present a 31-year-old man who, after a Conus textile sting acquired in New Caledonia, developed a cutaneous abscess on a buttock. The abscess was accompanied by pain, paraesthesia, general malaise, and fever. Complete remission was achieved by sodium hypochlorite packs and oral amoxicillin/clavulanic acid, metronidazole, and tramadol.