Analytical Quality by Design Assisted Optimization of RP-HPLC Method for the Estimation of Palovarotene Drug Substance and Drug Product by Box–Behnken Design

Introduction

Palovarotene (Sohonos, Ipsen, Paris, France) was approved by the USFDA on August 16, 2023, as a therapeutic option for fibrodysplasia ossificans progressive (FOP), after being approved in Canada in 2022. ¹ By binding to the retinoic acid receptor γ (RARγ), palovarotene suppresses BMP signaling; hence, chondrogenesis and, ultimately, HO is hindered. ² A bone morphogenetic protein (BMP) point mutation in the type I receptor is the cause of FOP, an incredibly rare autosomal dominant disorder. ³ Palovarotene is a selective agonist of the transcriptional repressor retinoic acid receptor gamma (RARγ), which is expressed in chondrocytes and chondrogenic cells. ¹ Palovarotene suppresses the SMAD1/5/8 signaling pathway by reducing BMP signaling by its binding to RARγ. ²

Palovarotene reduces muscle tissue damage by interfering with these pathways, which decreases chondrogenesis and permits normal muscle tissue repair. ³

No analytical techniques, including HPLC and UV, have been documented for the assessment of palovarotene in capsule formulation and API, and no studies on quality by design approach has been reported, according to literature reviews. Finding failure models and developing a reliable approach with adjustable design space for the duration of product life cycle management has been the main goal of AQbD⁴.

The present research aims to use analytical QbD principles to establish and optimize the RP-HPLC approach for estimating the capsule formulation of palovarotene. To achieve high resilience and improve method performance, the QbD concept has been widely used in the assessment of analytical parameters. ^5,6 QbD was discovered on the knowledge and application of (ICH Q8), (ICH Q9) and guidelines (ICH Q10). ^7,8

Materials And Methods

Chemicals

The Palovarotene API was gifted by MSN Labs Pvt., Ltd, Hyderabad, India. The marketed capsule formulation of Sohonos (Palovarotene 5 mg) was used for assay. Potassium dihydrogen phosphate & acetonitrile HPLC grade were purchased from Sigma Aldrich, Mumbai, India.

Instruments and Equipment

The technique was developed using a HPLC (Shimadzu) equipped system with a quaternary pump, a sonicator, sample, detector (PDA) (software) LC solution. Weighing machine (Shimadzu).

Stock solution (Standard)

Palovarotene 25 mg was measured in 25 ml VF and mixed with CH₃OH to prepare stock solution (1000μg/ml). By serially diluting this stock solution with methanol, several std solutions (10-60μg/ml) were enumerated. A nylon filter (0.45...