We delve into the challenges and solutions encountered during the method development, validation, and bioanalysis of complex molecules, specifically focusing on Liposomal-based formulations of Doxorubicin and Amphotericin B, as well as the Nanoparticle Albumin-bound Paclitaxel (nab-paclitaxel). Our experience and expertise in these areas have paved the way for successful projects and regulatory approvals.

Novel Drug Delivery Systems (NDDS) have revolutionized the pharmaceutical industry by offering advanced approaches to drug delivery. These systems aim to enhance bioavailability, minimize toxicity, and achieve controlled drug delivery, ensuring accurate dosing and improved patient safety and efficacy. NDDS enables site-specific drug delivery with optimized doses. Examples of NDDS include Microparticles, Nanoparticles, Liposomes, and more.

Liposomal based formulation of Doxorubicin and Amphotericin B

Quantification of encapsulated and un-encapsulated (Free) analyte in plasma
Challenges	Areas Identified to Control	Actions Taken
Protection of liposomal formulation from leakage On-site sample preparation logistic constraints Assay reproducibility and internal standard selection	Ensure formulation stability during sample collection and analysis Prevent mechanical damage to liposomes Implement sample collection with cryoprotective agents Develop extraction methods for free and encapsulated components Develop extraction methods for free and encapsulated components	Control sample temperature below 4 degrees Avoid rigorous shaking during sample handling Implement cryoprotective agents for sample collection Develop extraction methods for both components Utilize automated liquid handling for precision

Nanoparticle albumin-bound paclitaxel (nab-paclitaxel)

Unbound (Free) and Total Paclitaxel in plasma
Challenges	Areas Identified to Control	Actions Taken
Isolation of less than 10% of unbound Paclitaxel Inconsistent isolation process for free component Lot-to-lot assay reproducibility due to varying plasma binding Bench-top stability issues	Optimize isolation techniques without disturbing protein-bound analyte Reevaluate parameters involved in isolation for consistency Optimize extraction methods to compensate for different matrix protein binding Evaluate temperature conditions and stabilizers to address stability issues	Implement ultracentrifugation with kDa filters for isolation Optimize parameters such as incubation time, temperature, and centrifugation conditions for consistent results Adjust internal standard concentration to compensate for matrix protein binding Maintain temperature below 8 degrees.

Our Experience:

Doxorubicin – Liposomal

• Successfully completed 9 clinical trial projects
• Achieved regulatory approvals for 4 projects.
• Achieved over 90% acceptance for incurred samples reproducibility for both encapsulated and free analytes.
• Currently, one project is in its final stages, nearing completion for ANVISA submission

Amphotericin B – Liposomal

• Completed 2 projects for EU and US regulatory submissions.
• Achieved over 96% acceptance for incurred samples reproducibility for both encapsulated and free analytes.

Paclitaxel – Nanoparticle Albumin-bound

• Successfully completed 2 clinical trial projects for US regulatory submission.

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Lambda Therapeutic Research offers a wide range of bioanalytical capabilities specializing in cell-based assays, biomarkers, immunogenicity, and pharmacokinetics (PK). With state-of-the-art facilities and a team of experienced scientists, Lambda provides comprehensive solutions for the analysis of small molecules, biomarkers, biologics, and therapeutic trace elements. To know more, please visit https://www.lambda-cro.com/bio-analyticals.php