Introduction: In recent years, biuret derivatives have been studied due to their multiple biological effects, including analgesic[1], anti-inflammatory[2], anti-cancer[3] and etc. According to recent observations, the sedative effect[4] It was studied. However, due to the fact that some of the biuret derivatives are hydrophilic, their penetration through the blood-brain barrier and the emergence of their effects have become problematic or some other derivatives that are more lipophilic, the problem of dissolving them in water caused that after subcutaneous injection, their distribution in the body of the mice faced a problem and caused a weaker and delayed effect. In this study, the synthesis of such biuret derivatives and the preparation of amphiphilic nanoparticles from it to improve its pharmacokinetics and permeability and possibly improve its anticonvulsant effect.
Methods: 2.1. Synthesis of Biuret Derivative
By adding potassium cyanate (KOCN) to the hydrochloride solution made from amine ureas are made, which themselves react with phenyl chloroformate in the presence of a pyridine (pyr) catalyst, they create phenyl allophanate, which reacts with another amine under reflux conditions for one night in the presence of potassium carbonate (K2CO3) to produce the desired biuret.
2.2. Preparation of Nanoparticles from Biuret
Binary copolymer mPEG-PCL will be synthesized by ring-opening polymerization of caprolactone in the presence of polyethylene glycol as initiator molecule and tin octanate catalyst. Polymer nanoparticle will be prepared using nanoprecipitation method
2.3. In vivo Study of Anticonvulsant Effect
In this study, four groups of NMRI adult male mice weighing 25-30 grams were used. There are 10 mice in each group. Group I-control PTZ (35mg/kg) to cause chronic seizures, group II- positive control PTZ (35 mg/kg) + phenobarbital (20 mg/kg), group III- treatment group nano biuret derivative + PTZ (35 mg/kg), and group IV- control group biuret derivative (35 mg/kg) + PTZ (35 mg/kg). In all groups, the injection was done intraperitoneally, and PTZ was injected 30 minutes after phenobarbital and biuret.
In all groups, convulsive attacks are recorded 30 minutes after injection, and all abnormal animal behaviors are subclassified and scored, and finally, in order to check kindling, 10 days after the last injection, PTZ was re-injected at a dose of 75 mg/kg. and their behavior is checked for 30 minutes[5, 6].
2.4. Analysis of Nanoparticles
HNMR spectroscopy and FT-IR spectroscopy will be used to confirm and determine the structure of mPEG-PCL binary copolymer. Also, HNMR will be used to determine the average molecular mass and molecular mass distribution of mPEG-PCL binary copolymer, and DSC analysis will be used to study the thermal behavior of mPEG-PCL binary copolymer. A spectrophotometer will be used to study the amount of drug loading and drug release.
Results: The synthesis of biuret derivative was confirmed using UV/Vis, LC-Mass, IR. Also, by using DLS, TEM, IR the synthesis of nanoparticles was confirmed. According to similar articles, it was observed that biuret derivative micelles showed a better effect compared to biuret derivative itself.
Conclusion: According to previous studies and findings, the preparation of nanoparticles with mPEG-PCL copolymer can improve the pharmacokinetic and drug delivery effect of biuret derivatives, as a result, this technique can be used to improve the possible effect of other medicinal compounds with similar problems.
Keywords: biuret, nanoparticles, mPEG-PCL, drug delivery, anticonvulsant effect