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.

Sequence stratigraphy and sea-level history of Kangan formation in south pars field

Dehram group includes Faraghan, Dalan and Kangan formations. Kangan formation ages lower terias. That is one of the important reservoir rocks of southern Iran and Persian Gulf. In this research Kangan formation is studied in two A and B wells. Based on 75 studies on thin section, four carbonate litho acies association A, B, C, D with 12 subfacies are identified. A lithofacies association includes 4 subfacies: A1, A2, A3 and A4. B lithofacies association consists of 3 subfacies: B1, B2 and B3. C lithofacies association consists of 3 subfacies: C1, C2, C3 and D lithofacies association includes 2 subfacies: D1 and D2. On the base of studies lithofacies association of Kangan formations are formed in 3 environments of: Tidal Flat, Lagoon and Barrier Shore Complex in a Carbonated Platform Ramp type. Diagenetic processes have effected this formation. The most important Diagenetic processes are: Cementation, Anhydritization, Micrization, Neomorphism, Bioturbation, Dissolution, Compaction, Dolomitization and Porosity. Sequence staratigraphy studies were performed base on the vertical and horizontal relationship of lithofacies association and well logging in gamma ray and sonic type that causes the identification of two sedimentary sequences: First sedimentary sequence includes: Transgressive System Tract (TST) and High Stand System Tract (HST). The lower boundary of this sequence is in Sequence Boundary 1 (SB1) which shows unconformities of Dalan and Kangan that are Permian-terias unconformities. The upper boundary is in Sequence Boundary 2 (SB2) type that is identified by carbonate facies associated by anhydrite nodular. Second sedimentary sequence includes: TST and HST. Lower and upper boundaries of these sequences are both in SB2 type. The lower and upper boundary is made of carbonate facies with anhydrite nodular.