Author

Ritu Gupta

Document Type

Dissertation

Date of Award

8-2022

School/College

College of Pharmacy and Health Sciences (COPHS)

Degree Name

Ph.D. in Pharmaceutical Science

Committee Chairperson

Huan Xie

Committee Member 1

Dong Liang

Committee Member 2

Yuanjian Deng

Committee Member 3

Song Gao

Keywords

AC1LPSZG, Design of Experiments (DoE), In Vitro Drug Release, In Vivo Pharmacokinetics, PLGA, Preclinical Development

Abstract

Preclinical development of novel chemotherapeutic agent AC1LPSZG, a mammalian target of rapamycin (mTOR) inhibitor, involved development of sensitive reverse-phase ultra-performance liquid chromatography (UPLC) and LC-MS/MS methods for quantification of AC1LPSZG in in vitro study samples and rat plasma, respectively. Pharmacokinetic studies were done in SD rats after intravenous injection of cosolvent formulations. The resulting pharmacokinetic parameters were analyzed using non-compartmental analysis (NCA) and two-compartmental modeling. Poly (D, L-lactic-co-glycolic acid) (PLGA) is most used biodegradable synthetic polymer for nano drug delivery due to its non-toxic and biodegradable nature, and tunable release properties. PLGA nanoparticles (NPs) were prepared by ‘nanoprecipitation’ technique using a nonionic surfactant poloxamer P188. The particle size, size distribution, and zeta potential of prepared nanoparticles were analyzed using dynamic light scattering (DLS). The drug entrapment efficiency (%EE) was accessed by ultra-sonication of lyophilized NPs with acetonitrile and analyzing the drug content using UPLC. Design of Experiments (DoE) strategy using Design Expert® software (version 13) was successfully used to optimize PLGA (50:50) based NPs of AC1LPSZG. Optimized batch was prepared using 5 mg drug and 4 mL aqueous phase volume with EE of 41.2%, NP size of 124 nm, drug load of 2.6% and zeta potential of – 15 mV. We conclude similar DOE approaches can help to understand and optimize innovative manufacturing processes, needed for the quality by design (QbD) preparation of other nano-formulations. The in vitro drug release was tested in phosphate buffer pH 6.8 for 72 hours, employing USP-4 apparatus CE7-smart (SOTAX®) incorporated with Float-A-Lyzer dialysis cells at 300 kDa molecular weight cut–off (MWCO), flow rate 16 mL/min and temperature 37°C. Different surfactants were explored to enhance the drug solubility and accelerate the in vitro drug release. The influence of three different surfactants: SLS (Sodium Lauryl Sulfate-anionic), Tween 80 (non-ionic) and CTAB (Cetyltrimethylammonium bromide- cationic) on drug solubility, sink conditions and dissolution behavior was demonstrated. The solubility improvement was in the order of SLS > Tween80 > CTAB and dissolution efficiency was improved with the increase of surfactant concentration. The developed in vitro drug release method was able to discriminate among different release profiles. In brief, similar discriminatory test method can be used as a quality control tool to identify critical formulation and process parameters and can also be used as a surrogate for bioequivalence studies if a predictive IVIVC (In vitro In vivo correlation) is obtained.

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