Refolding of omega conotoxin peptide derived from marine snail, Conus magus, to evaluate its analgesic effects

Oxidative refolding is one of the key challenges hampering the development of peptide based compounds as therapeutics. The correct refolding for three disulfide peptide like w-Conotoxi n MVIIA is difficult and crucial for biological activity. This work advanced knowledge of chemical and biological for improve oxidative refolding of synthetic w-Conotoxi n MVIIA in base of Conus magus venom. The present study aimed to set up an appropriate and effective protocols for refolding of disulfide-rich w-Conotoxin MVIIA. In this study, the crude peptide was protected with Acm group, according to the right amino acid sequences (Synthesized by Australian Company). The crude peptide was purified by H PLC. To prepare the peptide to refolding, innovative deprotection applied molar ratio (AMR) method was performed based on mercury. Accuracy of deprotection was approved by reverse phase chromatography. The deprotected target peptide (omega-conotoxin) was determined by SDS-PAGE. Then the Oxidative refolding of target peptide was performed in six protocol based on Guanidinium chloride and oxidized and reduced Glutathione. Analgesic effect of refolded peptide was surveyed with formalin test in mice Balb/c. Non neurotoxic effects of target peptides were survey with ICV injection in mice model (C57/BL6). The innovative deprotection protocol performed based on the best ratio of mercury/2-mercaptoethanol adjusted to 1mg/10p1 in 90 minute. The results showed the yield and purity of omega-conotoxin MVIIA as 93 and 95%, respectively. Refolding of 40 mg omega Conotoxin with GSSG and GSH on ratio of 10:1 and 20 mM ammonium acetate showed the best analgesic effect compared with the other methods. The result showed 95.5% yield and 98% purity of omega-conotoxin MVIIA in this refolding method. Related refolding method reduced 85% pain in experimented mice using 7 ng of the peptide. That was 71.5 fold stronger than morphine and 2 times than standard Prialt®. And it was not neurotoxic in mice. In this study, refolding method for omega-conotoxin MVIIA was optimized in the fourth factor including: reducing the time, amount and number of reagent and increase the efficiency. We introduced new method for deprotection of omega-conotoxin MVIIA. Effective, economic and applied refolding and deprotecti on method was performed in this research may al so be applied to similar omega conotoxin peptides.

Design and construction of a two dimensional duct equipped with a wave generator and a variable coastal slope and an ocean wave energy conversion system (OWC)

The aims of this thesis were evaluation the type of wave channel, wave current, and effect of some parameters on them and identification and comparison between types of wave maker in laboratory situations. In this study, designing and making of two dimension channels (flume) and wave maker for experiment son the marine buoy, marine building and energy conversion systems were also investigated. In current research, the physical relation between pump and pumpage and the designing of current making in flume were evaluated. The related calculation for steel building, channels beside glasses and also equations of wave maker plate movement, power of motor and absorb wave(co astal slope) were calculated. In continue of this study, the servo motor was designed and applied for moving of wave maker’s plate. One Ball Screw Leaner was used for having better movement mechanisms of equipment and convert of the around movement to linear movement. The Programmable Logic Controller (PLC) was also used for control of wave maker system. The studies were explained type of ocean energies and energy conversion systems. In another part of this research, the systems of energy resistance in special way of Oscillating Water Column (OWC) were explained and one sample model was designed and applied in hydrolic channel at the Sheikh Bahaii building in Azad University, Science and Research Branch. The dimensions of designed flume was considered at 16 1.98 0. 57 m which had ability to provide regular waves as well as irregular waves with little changing on the control system. The ability of making waves was evaluated in our designed channel and the results were showed that all of the calculation in designed flume was correct. The mean of error between our results and theory calculation was conducted 7%, which was showed the well result in this situation. With evaluating of designed OWC model and considering of changes in the some part of system, one bigger sample of this model can be used for designing the energy conversion system model. The obtained results showed that the best form for chamber in exit position of system, were zero degree (0) in angle for moving below part, forty and five (45) degree in front wall of system and the moving forward of front wall keep in two times of height of wave.