摘要
Connexin43 mimetic peptide （Cx43MP） has been intensively investigated for its therapeutic effect in the management of inflammatory eye conditions, spinal cord injury, wound healing and iscbemia-induced brain damage. Here, we report on a validated stability-indicating reversed-phase high performance liquid chromatography （RP-HPLC） method for the quantification of Cx43MP under stress conditions. These included exposure to acid/base, light, oxidation and high temperature. In addition, the degradation kinetics of the peptide were evaluated in bovine vitreous and drug-free human plasma at 37℃. Detection of Cx43MP was carried out at 214 nm with a retention time of 7.5 min. The method showed excellent linearity over the concentration range of 0.9-250μg/mL （R2a0.998）, and the limits of detection （LOD） and quantification （LOQ） were found to be 0.90 and 2.98 μg/mL, respectively. The accuracy of the method determined by the mean percentage recovery at 7.8, 62.5 and 250μg/mL was 96.79%, 98.25% and 99.06% with a RSD of〈 2.2%. Accelerated stability studies revealed that Cx43MP was more sensitive to basic conditions and completely degraded within 24 h at 37℃（0% recovery） and within 12 h at 80℃ （0.34% recovery）. Cx43MP was found to be more stable in bovine vitreous （t1/2 slow= 171.8 min） compared to human plasma （t1/2slow = 39.3 min） at 37℃ according to the two phase degradation kinetic model. These findings are important for further pre-clinical development of Cx43MP.
Connexin43 mimetic peptide （Cx43MP） has been intensively investigated for its therapeutic effect in the management of inflammatory eye conditions, spinal cord injury, wound healing and iscbemia-induced brain damage. Here, we report on a validated stability-indicating reversed-phase high performance liquid chromatography （RP-HPLC） method for the quantification of Cx43MP under stress conditions. These included exposure to acid/base, light, oxidation and high temperature. In addition, the degradation kinetics of the peptide were evaluated in bovine vitreous and drug-free human plasma at 37℃. Detection of Cx43MP was carried out at 214 nm with a retention time of 7.5 min. The method showed excellent linearity over the concentration range of 0.9-250μg/mL （R2a0.998）, and the limits of detection （LOD） and quantification （LOQ） were found to be 0.90 and 2.98 μg/mL, respectively. The accuracy of the method determined by the mean percentage recovery at 7.8, 62.5 and 250μg/mL was 96.79%, 98.25% and 99.06% with a RSD of〈 2.2%. Accelerated stability studies revealed that Cx43MP was more sensitive to basic conditions and completely degraded within 24 h at 37℃（0% recovery） and within 12 h at 80℃ （0.34% recovery）. Cx43MP was found to be more stable in bovine vitreous （t1/2 slow= 171.8 min） compared to human plasma （t1/2slow = 39.3 min） at 37℃ according to the two phase degradation kinetic model. These findings are important for further pre-clinical development of Cx43MP.
基金
the University of Auckland for providing an international doctoral scholarship to Rohit Bisht.
