Abstracts for Posters 18-26
Poster 18
Quantifying the Intrinsic Effects of Lubrication to Predict the Tensile Strength of Pharmaceutical Formulations
Allan Hutchins, Matthew P. Mullarney
Pfizer, Inc.
Presenting Author: Allan Hutchins, [email protected]
Purpose
A model was developed to calculate the tensile strength of tablets compressed from a lubricated powder blend.
Methods
Conventional immediate release tablet diluents (microcrystalline cellulose, lactose, dibasic calcium phosphate) were each separately mixed with magnesium stearate at different levels and to different mixing extents to determine their individual lubrication sensitivity. Physical characteristics of each diluent that are formulation-specific were incorporated into a modification of the Kushner and Moore mixing sensitivity model and the Reynolds compressibility model to predict the system tensile strength.
Results
A good correlation was achieved between the measured and modeled tensile strength of over 50 different placebo blends.
Conclusion
The new model can be used to improve the design thinking for new direct compression tablet formulations and other predictive material models requiring the tensile strength of blends as an input.
Poster 19
Predictions of Drug Release in Bilayer Osmotic Tablets
Bhawana Singh Tomar (1), Weili Yu (1), Sweta Manthena (1), Alfred Berchielli (1), Kenneth Ogueri (1), Mahesh S Tirumkudulu (2), Pankaj Doshi (3)
(1) Pfizer Inc. Groton CT, USA (2) Department of Chemical Engineering, IIT Bombay, Mumbai, India, (3) Pfizer Products India Pvt. Ltd., Mumbai, India
Presenting Author: Bhawana Singh Tomar [email protected]
Corresponding Author: Mahesh S Tirumkudulu , [email protected]
Purpose
The Oral Osmotic pump (OROS) tablet, which controls drug release using osmotic pressure, offers several advantages over other drug delivery systems, including lower adverse reactions, improved compliance, reduced dosage, and independence from hydrodynamic conditions, gastric pH, and agitation. The first orally administered osmotic pump, the Extrudable Core System (ECS), provides easy preparation and near zero-order delivery of water-soluble drugs. However, ECS struggles with water-insoluble drugs due to incomplete extrusion once the osmotic agent is depleted. This issue is resolved by the bilayer system or swellable core technology (SCT), where the sweller layer ensures complete drug extrusion. This work aims to develop a mathematical model for SCT to aid formulators in designing better dosage forms.
Methods
We present a mechanistic, mathematical model describing drug release in a bilayer oral osmotic tablet coated with a semi-permeable membrane containing a single orifice. Drug release depends on several processes: solvent imbibition into the tablet, hydration of the solid core containing drug and polymer, hydration and swelling of the sweller layer containing polymer and osmogen, drug dispersibility, extrusion of hydrated components under hydrostatic pressure through the orifice, and dissolution of drug particles in bulk. Equations governing these processes are solved simultaneously to investigate the influence of various model parameters on the final release profile.
Results
We track changes in tablet dimensions (solid core, dispersed core, height of sweller layer, etc.), influx and outflux, hydrostatic pressure, osmotic pressure, tensile stress in the coating, changes in particle size distribution over time, and drug release. The model predicts trends in drug dissolved and other processes during dissolution as functions of coating permeability, drug dispersibility, and polymer properties. Our model is validated with observed release profiles of drugs with varying solubility, coating thickness, and drug loading.
Conclusion
Bilayer tablets (SCT) are preferred over single-layer osmotic tablets (ECS) due to the additional pressure generated by the sweller layer, resulting in higher drug release. The model was validated by comparing predictions with drug release data for three drugs of varying solubility, capturing the main trends observed in experiments, though further work is needed to refine the details.
Poster 20
Roller Compacted Ribbon Solid Fraction Measurement Methods Design Space
Amanda Strenk, Patrick Dente, Brent Harrington, Lina Niu
Pfizer, Inc.
Presenting Author: Amanda Strenk, [email protected]
Purpose
Roller compaction is a unit operation commonly used for solid dosage formulations to improve blend flowability and uniformity. During roller compaction, the powder blend is pressed into flat pieces of compacted blend (commonly called ribbons) between two rotating rollers and then milled against a screen to create granules. The ribbon solid fraction is a critical process parameter that needs to be accurately targeted to ensure favorable granule production for tablet compression.
Methods
Drug Product Manufacturing (DPM), the internal clinical manufacturing site for solid oral dosage forms at Pfizer, has been taking the initiative to study different methods to measure ribbon solid fraction with partner lines. In this study, throughput, punch tool, and Gasporox methods were evaluated to measure ribbon solid fraction. Throughput method measures ribbon solid fraction by weighing all ribbon output for a given time. However, in efforts to align across manufacturing sites, DPM began scoping punch tool method in 2022. Punch tool method varies from throughput method in that it involves direct individual ribbon dimensional analysis. Gasporox utilizes laser-based technologies to measure ribbon solid fraction. To further understand potential differences between these three methods and interactions between roller compaction factors such as roll force, gap width, roll speed, and roll design, DPM worked with partner lines to analyze the roller compaction design space. DPM has performed two designs of experiments using both placebo and active formulations to develop statistical models for each solid fraction measurement method in Design Expert.
Results
ANOVA results were analyzed for each model to determine significant factors and interactions. The results demonstrated that throughput method consistently has higher solid fraction results than the punch tool and Gasporox methods. This is likely due to the impact of fines bypass in throughput calculations and the estimation of ribbon thickness. Punch tool and Gasporox demonstrated the closest results out of the three methods. An additional finding is that fines bypass was positively correlated with gap width and roll speed.
Conclusion
DPM is continuing to leverage these results in defining and implementing a workflow for solid fraction measurements for roller compaction processing.
Poster 21
Improved microemulsion formation of clofazimine lipid digestion product-based SEDDS using linoleic acid
Nower Chowdhury, Hari P Kandagatla, and Abu T.M Serajuddin
St. John's University
Presenting Author: Hari P Kandagatla
Corresponding Author: Abu T.M Serajuddin, [email protected]
Purpose
This study investigates the enhanced emulsifying properties of long-chain polyunsaturated fatty acids containing two double bonds, focusing on how structural differences between linoleic acid (C18:2) and oleic acid (C18:1) influence their microemulsifying properties. We previously developed a lipid-based SEDDS for clofazimine (CFZ), an extremely water-insoluble antibiotic against leprosy and drug-resistant tuberculosis, using a lipid-digestion product (oleic acid plus glyceryl monooleate), which provided very high drug solubility. In this study, we investigated whether a lipid-digestion product containing linoleic plus glyceryl monolinoleate would also perform similarly or be better due to two double bonds.
Methods
Solubility studies of CFZ were conducted in formulation components, lipids, surfactants, and their combinations. SEDDS formulations with 10% drug loading were developed. In-vitro dispersion of the formulations was tested in various aqueous media, including an acidic medium (pH 2.0) and the phosphate buffer (pH 6.8) with and without a bile salt (sodium taurocholate).
Results
The solubility of CFZ in linoleic acid was very high (~480 mg/g) and comparable to that in oleic acid. A SEDDS formulation was developed using linoleic acid plus glyceryl monolinoleate (Maisine) as the lipid digestion product and a 1:1 Kolliphor EL-Tween 80 mixture as the surfactants, where the lipid-to-surfactant ratio was 1:1. The solubility of CFZ in the formulation was ~120 mg/g, and thus a 100 mg/g SEDDS was developed. In-vitro dispersion testing of the formulation containing 250 mL of media demonstrated complete dispersion at pH 6.8 medium with <250 nm particle size, but there was some crystallization of CFZ at pH 2 due to conversion to HCl salt and the common ion effect. Therefore, an enteric-coated capsule was used to overcome crystallization in gastric pH. Overall, the presence of linoleic acid and glyceryl monolinoleate in the formulation as a lipid-digestion product provided better performance than the oleic acid/oleate-based formulation.
Conclusion
A 1:1 mixture of glyceryl monolinoleate and linoleic acid significantly enhanced CFZ dispersibility. In-vitro testing showed pH-dependent solubility with crystallization at gastric pH and dissolution at intestinal pH. The crystallization challenge is addressed through the use of an enteric-coated capsule. This study demonstrates that CFZ exhibits high solubility in long-chain fatty acids, potentially eliminating the need for organic co-solvents in LBFs.
Poster 22
Impact of excipient purity on in vitro efficacy and safety of siRNA-loaded lipid nanoparticles
Sheyda Ranjbar and Xiuling Lu
University of Connecticut
Presenting Author: Sheyda Ranjbar, [email protected]
Corresponding Author: Xiuling Lu, [email protected]
Purpose
The quality of lipid components is crucial for the successful development of siRNA lipid nanoparticle (siRNA-LNP) therapeutics. This study examines how the vendor source and purity of D-Lin-MC3-DMA (MC3), a commonly used ionizable lipid, impact the critical quality attributes (CQAs) of siRNA-LNPs with a composition similar to the model drug Onpattro®. The findings offer a better scientific understanding on the effects of MC3 quality on in vitro efficacy and safety profile of siRNA-LNPs and provide valuable insights to facilitate the development of generic formulations.
Methods
siRNA-LNPs with three different sources of ionizable lipid MC3 were prepared using a microfluidic system and were dialyzed in phosphate-buffered saline. MC3 lipid purity was analyzed by liquid chromatography-mass spectrometry (LC-MS). Particle size distribution and surface charge were measured by zeta sizer. TNS binding assay was used to assess surface pKa. Encapsulation efficiency was measured using the Ribogreen assay. Cellular uptake and endosomal escape in HepG2 cells were evaluated by flow cytometry and confocal microscopy. Transthyretin (TTR) mRNA knockdown efficiency and cell viability in HepG2 cells were investigated by quantitative polymerase chain reaction (qPCR) and CellTiter-Glo® assay, respectively.
Results
Our in-house LC-MS analysis revealed varying purity levels (88–96%) of MC3 lipids procured from different vendors. The MC3 source investigated did not affect particle size, size distribution, surface pKa, zeta potential, or siRNA encapsulation efficiency. In contrast, higher purity level was associated with higher cellular uptake and endosomal escape. Nonetheless, gene silencing remained relatively unchanged at similar encapsulated siRNA concentrations, with EC₅₀ values around 1 nM. Furthermore, none of the formulations exhibited significant toxicity, even at concentrations 100 times higher than their EC₅₀ value.
Conclusion
The purity variations of MC3 lipids from different vendors affected cellular uptake and endosomal escape but showed no significant impact on gene silencing or cell viability in this study. Potential effects of ionizable lipid source on long-term stability and siRNA integrity is under investigation.
Poster 23
Nanoparticle-Mediated Inhalation Delivery of Celastrol for Targeted Therapy in non-small cell lung cancer (NSCLC)
Mural Quadros and Vivek Gupta
St. John's University
Presenting and Corresponding Author: Mural Quadros, [email protected]
Purpose
Non-small cell lung cancer (NSCLC) accounts for 85% of lung cancer cases and remains a leading cause of cancer-related mortality. Despite advancements in targeted therapies, the five-year survival rate for advanced NSCLC remains below 20%, highlighting the urgent need for novel treatments. Celastrol, a natural compound from Tripterygium wilfordii, exhibits anti-cancer, anti-inflammatory, anti-diabetic, and anti-microbial properties. However, its low aqueous solubility and poor bioavailability limit clinical application. Encapsulating celastrol in poly(lactic-co-glycolic acid) (PLGA) nanoparticles for inhalation delivery offers a strategy to improve solubility, bioavailability, and therapeutic efficacy. PLGA, a biocompatible and biodegradable polymer, provides controlled drug release, enhanced stability, and cellular uptake, making it an ideal delivery system.
Methods
Celastrol was encapsulated in PLGA nanoparticles using the solvent evaporation method with ~6 mg OSI, PLGA 50:50 (MW 10-15 kDa), 1% PVA, and 2 min sonication. The nanoparticles were characterized for drug encapsulation, particle size, and PDI. The anticancer activity of PLGA-Cela was evaluated in vitro using the MTT assay in H1975 NSCLC cells and compared to free celastrol after 72 h incubation.
Results
PLGA nanoparticles were successfully formulated with a drug encapsulation efficiency of 54.7 ± 11.8% and drug loading of 5.6 ± 1.2%. Physicochemical analysis showed particle size of 204.2 ± 1.3 nm, PDI of 0.01 ± 0.001, and surface charge of -26.30 ± 1.82 mV, ensuring uniformity and stability. In vitro assays demonstrated that PLGA-Cela exhibited significantly higher anticancer activity than free celastrol, with an IC₅₀ of 0.53 ± 0.07 µM versus 1.7 ± 0.07 µM for free celastrol.
Conclusion
This study successfully formulated celastrol-loaded PLGA nanoparticles for inhalation-based delivery in NSCLC treatment. The nanoparticles demonstrated high drug encapsulation and significantly enhanced anticancer activity compared to free celastrol. The improved cytotoxicity in H1975 cells suggests nanoparticle-mediated delivery could be a promising strategy for treating NSCLC. Further studies in 2D and 3D cell culture models and aerosolization assessments are being conducted to establish the potential of this formulation for effective lung cancer therapy.
Poster 24
A Nano-emulsion Formulation for Improved Oral Bioavailability of Sorafenib, and Potential Repurposing for Lung Cancer Treatment
Sanjana Durve, Dyandev Gadhave, Vivek Gupta
St. John's University
Presenting Author: Sanjana Durve, [email protected]
Corresponding Author: Vivek Gupta, [email protected]
Purpose
Lung cancer remains a leading cause of cancer related deaths in the U.S, with Non-Small Cell Lung Cancer (NSCLC) being the primary driver of cancer-related deaths. Current treatments, include chemotherapy, targeted therapy, and immunotherapy, face limitations such as drug resistance, toxicity, high cost, and systemic drug exposure. Repurposing kinase inhibitors like Sorafenib (SF) offers a promising approach. Our research previously demonstrated Sorafenib’s efficacy against NSCLC via polymeric nanoparticles. Since Sorafenib is a BCS class II drug with limited bioavailability, this study aims to develop an oral nano-emulsion (SF-NE) to enhance Sorafenib’s solubility and bioavailability.
Methods
SF-NE was prepared using low-energy and water titration methods with Capmul MCM (oil), Tween-20 (surfactant), and Transcutol HP (co-surfactant). Pseudo-ternary phase diagrams with different surfactant-to-co-surfactant (Smix) ratios (1:1, 1:2, 2:1, and 3:1) determined the optimal composition. Sorafenib was dissolved in Capmul MCM at 70°C and mixed at 1,000 rpm. Subsequently, Smix was incorporated in 3:1 ratio with gradual addition of water to achieve a stable, homogeneous nano-emulsion. The formulation was characterized for particle size, zeta potential, polydispersity index (PDI), and entrapment efficiency. Particle morphology was observed using Transmission electron microscopy (TEM). SF-NE’s efficacy on NSCLC cell lines and permeability on Caco-2 cells will be evaluated.
Results
Pseudo-ternary diagrams identified the (Smix 3:1) ratio as optimal. Among eight formulations, optimized SF-NE (T-8) exhibited a particle size of 41.6±22.7nm, PDI of 0.4±0.1, zeta potential of –4.7±2.01 mV, and entrapment efficiency of 88.0±5.7%. TEM confirmed the spherical particle morphology. Stability testing, freeze-thaw cycles and centrifugation demonstrated no phase separation, The cytotoxicity study revealed that there was ~10-fold reduction in IC50 value of SF-NE than plane drug on A549 cell line. Furthermore, qualitative images of cellular internalization showed more fluorescence in SF-NE compared to drug.
Conclusion
This study demonstrates the feasibility of SF-NE against NSCLC with particle size <100 nm, high drug encapsulation (>80%), and excellent thermodynamic stability. Further studies will investigate in-vitro drug release and the potential of SF-NE in 3D tumor model of NSCLC. In-vitro permeability studies using Caco-2 cells will be conducted to assess in-vivo oral absorption of the SF-Nano-emulsion.
Poster 25
Inhalable Liposomal Clofazimine for Targeted Pleural Mesothelioma treatment
Suman Choudhary, Mimansa Goyal, Vivek Gupta
St. John's University
Presenting Author: Suman Choudhary, [email protected]
Corresponding Authors: Vivek Gupta, [email protected], and Mimansa Goyal, [email protected]
Purpose
Malignant Pleural Mesothelioma (MPM) is a rare and aggressive type of cancer occurring in the pleural lining around the lungs. MPM has been primarily linked to exposure to aerosolized asbestos, a major issue in war veterans, construction workers and first responders, with a recent mass exposure during 9/11 collapse of World trade center towers. The prolonged latency period complicates early diagnosis and contributes to a dismal prognosis with a meager survival rate (<10%) and average life expectancy of a year. Our research lab previously reported effective repurposing of Clofazimine (CFZ), an anti-tuberculosis drug, for MPM treatment. CFZ was shown to inhibit autophagy and induce apoptosis in MPM cells. In this project, we aim to develop an inhaled liposomal formulation of CFZ to improve deep lung deposition and provide targeted effect with minimum systemic exposure at a reduced dose.
Methods
CFZ liposomes were prepared via conventional thin-film hydration method using DPPC and Cholesterol (Molar ratio 7:3) with varying total lipid content and CFZ amount (F1-F5). Formulations were characterized for entrapment, drug loading, particle size, PDI, and zeta potential. Morphology was studied using TEM. Solid state characterization was carried out using DSC and XRD. In-vitro drug release was performed to study the release pattern and aerodynamic performance were evaluated using NGI. In vitro cell culture studies and cellular internalization were performed on immortalized MPM cell line. Stability testing was performed at different storage conditions.
Results
CFZ was efficiently loaded in liposomes (F5) with higher drug loading of 7.8±0.1%. DSC and XRD confirmed CFZ entrapment. F5 formulation demonstrated stability when stored at 4oC. In-vitro release showed a burst release of ~60% drug release at 6 h in PBS containing 1% Tween 80. F5 showed deep lung deposition with MMAD of 3.4±0.1 µm and FPF of 81.5±1.1%. The cytotoxic studies showed ~5.6-fold reduction in IC50 for CFZ-liposomes versus CFZ in MSTO-211H cell line, while blank liposomes were non-toxic to L2 cell line. Fluorescent images revealed enhanced internalization of the formulation in cancer cells, with quantified results showing approximately two-fold higher uptake.
Conclusion
CFZ liposomal formulation demonstrated efficient drug loading, sustained release, deep lung deposition, enhanced cellular uptake, suggesting its potential for targeted MPM treatment via inhalation. Ongoing studies include 2D/3D in vitro assays on various MPM cell lines.
Poster 26
Impact of Particle Size Reduction Technique on the Performance of Long-Acting Injectable Suspensions
Saurabh Bhorkade and Diane Burgess
University of Connecticut
Presenting Author: Saurabh Bhorkade, [email protected]
Corresponding Author: Diane Burgess, [email protected]
Purpose
Long-acting injectable (LAI) suspensions offer significant therapeutic potential. However, these formulations are not fully understood due to their complex nature. This study examines how various particle size reduction methods affect physicochemical properties and in vitro release of medroxyprogesterone acetate (MPA) LAI suspensions. The goal is to elucidate the relationship between particle characteristics and formulation performance.
Methods
Three techniques (antisolvent recrystallization, bead milling, and jet milling) were used to prepare MPA crystals of similar mean particle size (small and large). Thermal characterization of the prepared API was conducted using DSC and TGA to detect residual solvent and polymorphic changes. PXRD analysis was performed to assess crystallinity, PLM and SEM were used to observe morphology. Q1/Q2 equivalent formulations of DepoProvera 150 were prepared using the various MPA crystals and characterized for particle size and distribution. In vitro release testing was carried out using USP dissolution apparatus IV with in-house adapters and 1% SDS release media at 37°C.
Results
MPA crystals with similar smaller (3 μm) and larger (20 μm) particle size were successfully obtained. Thermal analysis and PXRD confirmed the crystalline nature and absence of polymorphic changes. Particle size analysis of Q1/Q2 equivalent formulations showed consistent results, with D50 of 3 μm for small particle size formulations (except jet-milled API, D50 ≈ 7μm) and D50 of 20 μm for large particle size formulations. In vitro release studies revealed faster release for the jet-milled API formulation compared to other small particle size formulations. SEM imaging indicated that jet-milled API particles were present as aggregates with actual sizes smaller than 3 μm, explaining the accelerated release. Similarly, formulations with larger particle size solvent-recrystallized API exhibited faster than expected release, which was attributed to their distinct morphology resulting in tightly bound aggregates.
Conclusion
This study demonstrates that particle size reduction techniques can significantly impact in vitro release and overall performance of LAI suspensions. The findings highlight the importance of considering not only mean particle size but also the morphology and aggregation state of the API particles when developing and optimizing LAI formulations. These findings will help formulators choose effective particle size reduction methods and help predict release behavior.
Poster 18
Quantifying the Intrinsic Effects of Lubrication to Predict the Tensile Strength of Pharmaceutical Formulations
Allan Hutchins, Matthew P. Mullarney
Pfizer, Inc.
Presenting Author: Allan Hutchins, [email protected]
Purpose
A model was developed to calculate the tensile strength of tablets compressed from a lubricated powder blend.
Methods
Conventional immediate release tablet diluents (microcrystalline cellulose, lactose, dibasic calcium phosphate) were each separately mixed with magnesium stearate at different levels and to different mixing extents to determine their individual lubrication sensitivity. Physical characteristics of each diluent that are formulation-specific were incorporated into a modification of the Kushner and Moore mixing sensitivity model and the Reynolds compressibility model to predict the system tensile strength.
Results
A good correlation was achieved between the measured and modeled tensile strength of over 50 different placebo blends.
Conclusion
The new model can be used to improve the design thinking for new direct compression tablet formulations and other predictive material models requiring the tensile strength of blends as an input.
Poster 19
Predictions of Drug Release in Bilayer Osmotic Tablets
Bhawana Singh Tomar (1), Weili Yu (1), Sweta Manthena (1), Alfred Berchielli (1), Kenneth Ogueri (1), Mahesh S Tirumkudulu (2), Pankaj Doshi (3)
(1) Pfizer Inc. Groton CT, USA (2) Department of Chemical Engineering, IIT Bombay, Mumbai, India, (3) Pfizer Products India Pvt. Ltd., Mumbai, India
Presenting Author: Bhawana Singh Tomar [email protected]
Corresponding Author: Mahesh S Tirumkudulu , [email protected]
Purpose
The Oral Osmotic pump (OROS) tablet, which controls drug release using osmotic pressure, offers several advantages over other drug delivery systems, including lower adverse reactions, improved compliance, reduced dosage, and independence from hydrodynamic conditions, gastric pH, and agitation. The first orally administered osmotic pump, the Extrudable Core System (ECS), provides easy preparation and near zero-order delivery of water-soluble drugs. However, ECS struggles with water-insoluble drugs due to incomplete extrusion once the osmotic agent is depleted. This issue is resolved by the bilayer system or swellable core technology (SCT), where the sweller layer ensures complete drug extrusion. This work aims to develop a mathematical model for SCT to aid formulators in designing better dosage forms.
Methods
We present a mechanistic, mathematical model describing drug release in a bilayer oral osmotic tablet coated with a semi-permeable membrane containing a single orifice. Drug release depends on several processes: solvent imbibition into the tablet, hydration of the solid core containing drug and polymer, hydration and swelling of the sweller layer containing polymer and osmogen, drug dispersibility, extrusion of hydrated components under hydrostatic pressure through the orifice, and dissolution of drug particles in bulk. Equations governing these processes are solved simultaneously to investigate the influence of various model parameters on the final release profile.
Results
We track changes in tablet dimensions (solid core, dispersed core, height of sweller layer, etc.), influx and outflux, hydrostatic pressure, osmotic pressure, tensile stress in the coating, changes in particle size distribution over time, and drug release. The model predicts trends in drug dissolved and other processes during dissolution as functions of coating permeability, drug dispersibility, and polymer properties. Our model is validated with observed release profiles of drugs with varying solubility, coating thickness, and drug loading.
Conclusion
Bilayer tablets (SCT) are preferred over single-layer osmotic tablets (ECS) due to the additional pressure generated by the sweller layer, resulting in higher drug release. The model was validated by comparing predictions with drug release data for three drugs of varying solubility, capturing the main trends observed in experiments, though further work is needed to refine the details.
Poster 20
Roller Compacted Ribbon Solid Fraction Measurement Methods Design Space
Amanda Strenk, Patrick Dente, Brent Harrington, Lina Niu
Pfizer, Inc.
Presenting Author: Amanda Strenk, [email protected]
Purpose
Roller compaction is a unit operation commonly used for solid dosage formulations to improve blend flowability and uniformity. During roller compaction, the powder blend is pressed into flat pieces of compacted blend (commonly called ribbons) between two rotating rollers and then milled against a screen to create granules. The ribbon solid fraction is a critical process parameter that needs to be accurately targeted to ensure favorable granule production for tablet compression.
Methods
Drug Product Manufacturing (DPM), the internal clinical manufacturing site for solid oral dosage forms at Pfizer, has been taking the initiative to study different methods to measure ribbon solid fraction with partner lines. In this study, throughput, punch tool, and Gasporox methods were evaluated to measure ribbon solid fraction. Throughput method measures ribbon solid fraction by weighing all ribbon output for a given time. However, in efforts to align across manufacturing sites, DPM began scoping punch tool method in 2022. Punch tool method varies from throughput method in that it involves direct individual ribbon dimensional analysis. Gasporox utilizes laser-based technologies to measure ribbon solid fraction. To further understand potential differences between these three methods and interactions between roller compaction factors such as roll force, gap width, roll speed, and roll design, DPM worked with partner lines to analyze the roller compaction design space. DPM has performed two designs of experiments using both placebo and active formulations to develop statistical models for each solid fraction measurement method in Design Expert.
Results
ANOVA results were analyzed for each model to determine significant factors and interactions. The results demonstrated that throughput method consistently has higher solid fraction results than the punch tool and Gasporox methods. This is likely due to the impact of fines bypass in throughput calculations and the estimation of ribbon thickness. Punch tool and Gasporox demonstrated the closest results out of the three methods. An additional finding is that fines bypass was positively correlated with gap width and roll speed.
Conclusion
DPM is continuing to leverage these results in defining and implementing a workflow for solid fraction measurements for roller compaction processing.
Poster 21
Improved microemulsion formation of clofazimine lipid digestion product-based SEDDS using linoleic acid
Nower Chowdhury, Hari P Kandagatla, and Abu T.M Serajuddin
St. John's University
Presenting Author: Hari P Kandagatla
Corresponding Author: Abu T.M Serajuddin, [email protected]
Purpose
This study investigates the enhanced emulsifying properties of long-chain polyunsaturated fatty acids containing two double bonds, focusing on how structural differences between linoleic acid (C18:2) and oleic acid (C18:1) influence their microemulsifying properties. We previously developed a lipid-based SEDDS for clofazimine (CFZ), an extremely water-insoluble antibiotic against leprosy and drug-resistant tuberculosis, using a lipid-digestion product (oleic acid plus glyceryl monooleate), which provided very high drug solubility. In this study, we investigated whether a lipid-digestion product containing linoleic plus glyceryl monolinoleate would also perform similarly or be better due to two double bonds.
Methods
Solubility studies of CFZ were conducted in formulation components, lipids, surfactants, and their combinations. SEDDS formulations with 10% drug loading were developed. In-vitro dispersion of the formulations was tested in various aqueous media, including an acidic medium (pH 2.0) and the phosphate buffer (pH 6.8) with and without a bile salt (sodium taurocholate).
Results
The solubility of CFZ in linoleic acid was very high (~480 mg/g) and comparable to that in oleic acid. A SEDDS formulation was developed using linoleic acid plus glyceryl monolinoleate (Maisine) as the lipid digestion product and a 1:1 Kolliphor EL-Tween 80 mixture as the surfactants, where the lipid-to-surfactant ratio was 1:1. The solubility of CFZ in the formulation was ~120 mg/g, and thus a 100 mg/g SEDDS was developed. In-vitro dispersion testing of the formulation containing 250 mL of media demonstrated complete dispersion at pH 6.8 medium with <250 nm particle size, but there was some crystallization of CFZ at pH 2 due to conversion to HCl salt and the common ion effect. Therefore, an enteric-coated capsule was used to overcome crystallization in gastric pH. Overall, the presence of linoleic acid and glyceryl monolinoleate in the formulation as a lipid-digestion product provided better performance than the oleic acid/oleate-based formulation.
Conclusion
A 1:1 mixture of glyceryl monolinoleate and linoleic acid significantly enhanced CFZ dispersibility. In-vitro testing showed pH-dependent solubility with crystallization at gastric pH and dissolution at intestinal pH. The crystallization challenge is addressed through the use of an enteric-coated capsule. This study demonstrates that CFZ exhibits high solubility in long-chain fatty acids, potentially eliminating the need for organic co-solvents in LBFs.
Poster 22
Impact of excipient purity on in vitro efficacy and safety of siRNA-loaded lipid nanoparticles
Sheyda Ranjbar and Xiuling Lu
University of Connecticut
Presenting Author: Sheyda Ranjbar, [email protected]
Corresponding Author: Xiuling Lu, [email protected]
Purpose
The quality of lipid components is crucial for the successful development of siRNA lipid nanoparticle (siRNA-LNP) therapeutics. This study examines how the vendor source and purity of D-Lin-MC3-DMA (MC3), a commonly used ionizable lipid, impact the critical quality attributes (CQAs) of siRNA-LNPs with a composition similar to the model drug Onpattro®. The findings offer a better scientific understanding on the effects of MC3 quality on in vitro efficacy and safety profile of siRNA-LNPs and provide valuable insights to facilitate the development of generic formulations.
Methods
siRNA-LNPs with three different sources of ionizable lipid MC3 were prepared using a microfluidic system and were dialyzed in phosphate-buffered saline. MC3 lipid purity was analyzed by liquid chromatography-mass spectrometry (LC-MS). Particle size distribution and surface charge were measured by zeta sizer. TNS binding assay was used to assess surface pKa. Encapsulation efficiency was measured using the Ribogreen assay. Cellular uptake and endosomal escape in HepG2 cells were evaluated by flow cytometry and confocal microscopy. Transthyretin (TTR) mRNA knockdown efficiency and cell viability in HepG2 cells were investigated by quantitative polymerase chain reaction (qPCR) and CellTiter-Glo® assay, respectively.
Results
Our in-house LC-MS analysis revealed varying purity levels (88–96%) of MC3 lipids procured from different vendors. The MC3 source investigated did not affect particle size, size distribution, surface pKa, zeta potential, or siRNA encapsulation efficiency. In contrast, higher purity level was associated with higher cellular uptake and endosomal escape. Nonetheless, gene silencing remained relatively unchanged at similar encapsulated siRNA concentrations, with EC₅₀ values around 1 nM. Furthermore, none of the formulations exhibited significant toxicity, even at concentrations 100 times higher than their EC₅₀ value.
Conclusion
The purity variations of MC3 lipids from different vendors affected cellular uptake and endosomal escape but showed no significant impact on gene silencing or cell viability in this study. Potential effects of ionizable lipid source on long-term stability and siRNA integrity is under investigation.
Poster 23
Nanoparticle-Mediated Inhalation Delivery of Celastrol for Targeted Therapy in non-small cell lung cancer (NSCLC)
Mural Quadros and Vivek Gupta
St. John's University
Presenting and Corresponding Author: Mural Quadros, [email protected]
Purpose
Non-small cell lung cancer (NSCLC) accounts for 85% of lung cancer cases and remains a leading cause of cancer-related mortality. Despite advancements in targeted therapies, the five-year survival rate for advanced NSCLC remains below 20%, highlighting the urgent need for novel treatments. Celastrol, a natural compound from Tripterygium wilfordii, exhibits anti-cancer, anti-inflammatory, anti-diabetic, and anti-microbial properties. However, its low aqueous solubility and poor bioavailability limit clinical application. Encapsulating celastrol in poly(lactic-co-glycolic acid) (PLGA) nanoparticles for inhalation delivery offers a strategy to improve solubility, bioavailability, and therapeutic efficacy. PLGA, a biocompatible and biodegradable polymer, provides controlled drug release, enhanced stability, and cellular uptake, making it an ideal delivery system.
Methods
Celastrol was encapsulated in PLGA nanoparticles using the solvent evaporation method with ~6 mg OSI, PLGA 50:50 (MW 10-15 kDa), 1% PVA, and 2 min sonication. The nanoparticles were characterized for drug encapsulation, particle size, and PDI. The anticancer activity of PLGA-Cela was evaluated in vitro using the MTT assay in H1975 NSCLC cells and compared to free celastrol after 72 h incubation.
Results
PLGA nanoparticles were successfully formulated with a drug encapsulation efficiency of 54.7 ± 11.8% and drug loading of 5.6 ± 1.2%. Physicochemical analysis showed particle size of 204.2 ± 1.3 nm, PDI of 0.01 ± 0.001, and surface charge of -26.30 ± 1.82 mV, ensuring uniformity and stability. In vitro assays demonstrated that PLGA-Cela exhibited significantly higher anticancer activity than free celastrol, with an IC₅₀ of 0.53 ± 0.07 µM versus 1.7 ± 0.07 µM for free celastrol.
Conclusion
This study successfully formulated celastrol-loaded PLGA nanoparticles for inhalation-based delivery in NSCLC treatment. The nanoparticles demonstrated high drug encapsulation and significantly enhanced anticancer activity compared to free celastrol. The improved cytotoxicity in H1975 cells suggests nanoparticle-mediated delivery could be a promising strategy for treating NSCLC. Further studies in 2D and 3D cell culture models and aerosolization assessments are being conducted to establish the potential of this formulation for effective lung cancer therapy.
Poster 24
A Nano-emulsion Formulation for Improved Oral Bioavailability of Sorafenib, and Potential Repurposing for Lung Cancer Treatment
Sanjana Durve, Dyandev Gadhave, Vivek Gupta
St. John's University
Presenting Author: Sanjana Durve, [email protected]
Corresponding Author: Vivek Gupta, [email protected]
Purpose
Lung cancer remains a leading cause of cancer related deaths in the U.S, with Non-Small Cell Lung Cancer (NSCLC) being the primary driver of cancer-related deaths. Current treatments, include chemotherapy, targeted therapy, and immunotherapy, face limitations such as drug resistance, toxicity, high cost, and systemic drug exposure. Repurposing kinase inhibitors like Sorafenib (SF) offers a promising approach. Our research previously demonstrated Sorafenib’s efficacy against NSCLC via polymeric nanoparticles. Since Sorafenib is a BCS class II drug with limited bioavailability, this study aims to develop an oral nano-emulsion (SF-NE) to enhance Sorafenib’s solubility and bioavailability.
Methods
SF-NE was prepared using low-energy and water titration methods with Capmul MCM (oil), Tween-20 (surfactant), and Transcutol HP (co-surfactant). Pseudo-ternary phase diagrams with different surfactant-to-co-surfactant (Smix) ratios (1:1, 1:2, 2:1, and 3:1) determined the optimal composition. Sorafenib was dissolved in Capmul MCM at 70°C and mixed at 1,000 rpm. Subsequently, Smix was incorporated in 3:1 ratio with gradual addition of water to achieve a stable, homogeneous nano-emulsion. The formulation was characterized for particle size, zeta potential, polydispersity index (PDI), and entrapment efficiency. Particle morphology was observed using Transmission electron microscopy (TEM). SF-NE’s efficacy on NSCLC cell lines and permeability on Caco-2 cells will be evaluated.
Results
Pseudo-ternary diagrams identified the (Smix 3:1) ratio as optimal. Among eight formulations, optimized SF-NE (T-8) exhibited a particle size of 41.6±22.7nm, PDI of 0.4±0.1, zeta potential of –4.7±2.01 mV, and entrapment efficiency of 88.0±5.7%. TEM confirmed the spherical particle morphology. Stability testing, freeze-thaw cycles and centrifugation demonstrated no phase separation, The cytotoxicity study revealed that there was ~10-fold reduction in IC50 value of SF-NE than plane drug on A549 cell line. Furthermore, qualitative images of cellular internalization showed more fluorescence in SF-NE compared to drug.
Conclusion
This study demonstrates the feasibility of SF-NE against NSCLC with particle size <100 nm, high drug encapsulation (>80%), and excellent thermodynamic stability. Further studies will investigate in-vitro drug release and the potential of SF-NE in 3D tumor model of NSCLC. In-vitro permeability studies using Caco-2 cells will be conducted to assess in-vivo oral absorption of the SF-Nano-emulsion.
Poster 25
Inhalable Liposomal Clofazimine for Targeted Pleural Mesothelioma treatment
Suman Choudhary, Mimansa Goyal, Vivek Gupta
St. John's University
Presenting Author: Suman Choudhary, [email protected]
Corresponding Authors: Vivek Gupta, [email protected], and Mimansa Goyal, [email protected]
Purpose
Malignant Pleural Mesothelioma (MPM) is a rare and aggressive type of cancer occurring in the pleural lining around the lungs. MPM has been primarily linked to exposure to aerosolized asbestos, a major issue in war veterans, construction workers and first responders, with a recent mass exposure during 9/11 collapse of World trade center towers. The prolonged latency period complicates early diagnosis and contributes to a dismal prognosis with a meager survival rate (<10%) and average life expectancy of a year. Our research lab previously reported effective repurposing of Clofazimine (CFZ), an anti-tuberculosis drug, for MPM treatment. CFZ was shown to inhibit autophagy and induce apoptosis in MPM cells. In this project, we aim to develop an inhaled liposomal formulation of CFZ to improve deep lung deposition and provide targeted effect with minimum systemic exposure at a reduced dose.
Methods
CFZ liposomes were prepared via conventional thin-film hydration method using DPPC and Cholesterol (Molar ratio 7:3) with varying total lipid content and CFZ amount (F1-F5). Formulations were characterized for entrapment, drug loading, particle size, PDI, and zeta potential. Morphology was studied using TEM. Solid state characterization was carried out using DSC and XRD. In-vitro drug release was performed to study the release pattern and aerodynamic performance were evaluated using NGI. In vitro cell culture studies and cellular internalization were performed on immortalized MPM cell line. Stability testing was performed at different storage conditions.
Results
CFZ was efficiently loaded in liposomes (F5) with higher drug loading of 7.8±0.1%. DSC and XRD confirmed CFZ entrapment. F5 formulation demonstrated stability when stored at 4oC. In-vitro release showed a burst release of ~60% drug release at 6 h in PBS containing 1% Tween 80. F5 showed deep lung deposition with MMAD of 3.4±0.1 µm and FPF of 81.5±1.1%. The cytotoxic studies showed ~5.6-fold reduction in IC50 for CFZ-liposomes versus CFZ in MSTO-211H cell line, while blank liposomes were non-toxic to L2 cell line. Fluorescent images revealed enhanced internalization of the formulation in cancer cells, with quantified results showing approximately two-fold higher uptake.
Conclusion
CFZ liposomal formulation demonstrated efficient drug loading, sustained release, deep lung deposition, enhanced cellular uptake, suggesting its potential for targeted MPM treatment via inhalation. Ongoing studies include 2D/3D in vitro assays on various MPM cell lines.
Poster 26
Impact of Particle Size Reduction Technique on the Performance of Long-Acting Injectable Suspensions
Saurabh Bhorkade and Diane Burgess
University of Connecticut
Presenting Author: Saurabh Bhorkade, [email protected]
Corresponding Author: Diane Burgess, [email protected]
Purpose
Long-acting injectable (LAI) suspensions offer significant therapeutic potential. However, these formulations are not fully understood due to their complex nature. This study examines how various particle size reduction methods affect physicochemical properties and in vitro release of medroxyprogesterone acetate (MPA) LAI suspensions. The goal is to elucidate the relationship between particle characteristics and formulation performance.
Methods
Three techniques (antisolvent recrystallization, bead milling, and jet milling) were used to prepare MPA crystals of similar mean particle size (small and large). Thermal characterization of the prepared API was conducted using DSC and TGA to detect residual solvent and polymorphic changes. PXRD analysis was performed to assess crystallinity, PLM and SEM were used to observe morphology. Q1/Q2 equivalent formulations of DepoProvera 150 were prepared using the various MPA crystals and characterized for particle size and distribution. In vitro release testing was carried out using USP dissolution apparatus IV with in-house adapters and 1% SDS release media at 37°C.
Results
MPA crystals with similar smaller (3 μm) and larger (20 μm) particle size were successfully obtained. Thermal analysis and PXRD confirmed the crystalline nature and absence of polymorphic changes. Particle size analysis of Q1/Q2 equivalent formulations showed consistent results, with D50 of 3 μm for small particle size formulations (except jet-milled API, D50 ≈ 7μm) and D50 of 20 μm for large particle size formulations. In vitro release studies revealed faster release for the jet-milled API formulation compared to other small particle size formulations. SEM imaging indicated that jet-milled API particles were present as aggregates with actual sizes smaller than 3 μm, explaining the accelerated release. Similarly, formulations with larger particle size solvent-recrystallized API exhibited faster than expected release, which was attributed to their distinct morphology resulting in tightly bound aggregates.
Conclusion
This study demonstrates that particle size reduction techniques can significantly impact in vitro release and overall performance of LAI suspensions. The findings highlight the importance of considering not only mean particle size but also the morphology and aggregation state of the API particles when developing and optimizing LAI formulations. These findings will help formulators choose effective particle size reduction methods and help predict release behavior.
