Utilization of In Vitro, In Vivo and In Silico Tools to Evaluate the pH-Dependent Absorption of a BCS Class II Compound and Identify a pH-Effect Mitigating Strategy.

PURPOSE: To describe a stepwise approach to evaluate the pH effect for a weakly basic drug by in vitro, in vivo and in silico techniques and identify a viable mitigation strategy that addresses the risk. METHODS: Clinical studies included assessment of the pH effect with famotidine. In vitro dissolution was evaluated in various biorelevant media and in a pH-shift test. PK studies in dogs were conducted under pentagastrin or famotidine pre-treatment and GastroPlus was employed to model human and dog PK data and simulate the performance in human. RESULTS: Clinical data indicated considerable pH dependent absorption of the drug when dosed in the presence of H2-antagonists. In vitro dissolution and in vivo dog data confirmed that the observed pH effect was due to reduced dissolution rate and lower solubility at increased gastric and intestinal pH. A salt form was identified to overcome the effect by providing fast dissolution and prolonged supersaturation. GastroPlus simulations predicted a mitigation of the pH effect by the salt. CONCLUSIONS: The drug exhibited a strong pH-effect in humans. The in vitro, in vivo and modeling approach provides a systematic workflow to evaluate the risk of a new drug and identify a strategy able to mitigate the risk.

A New Device for Securing Sternal Wires After Median Sternotomy: Biomechanical Study and Retrospective Clinical Assessment.

OBJECTIVE: Morbidity due to sternotomy continues to be a significant clinical problem. Poor approximation of the sternum may lead to complications such as sternal dehiscence, infection, and pain. A device to assist in tensioning and twisting standard steel wires during sternal closure has been developed (TORQ sternal closure device). Manually tightened interrupted wire closures were compared with those tightened and secured with the aid of the device. Performance of the device was assessed clinically. METHODS: Four cardiovascular surgeons performed manual and device-assisted closures on a biofidelic model. Closure force was measured to determine the residual force and its intraoperator variation. A retrospective review of patients treated before and after the introduction of the device was conducted. Predicted and actual outcomes were compared for the two groups (manual closure and device-assisted closure). RESULTS: Biomechanical testing measured a 75% increase in residual closure force (P < 0.001) and a significant reduction in the variability of the closure force (P = 0.045) for device-assisted closures compared with manual closures. In the retrospective study, 3 of 173 manually closed patients had sterile sternal dehiscence and 1 of 173 had a deep sternal wound infection. In the device closure group, 2 of 127 had a sterile sternal dehiscence and no deep sternal wound infections were reported. No other device-related serious adverse events were reported. CONCLUSIONS: Biomechanical data showed stronger, more consistent closure forces with the device. The retrospective data attest to the performance of the device.

Food Effect in Humans: Predicting the Risk Through In Vitro Dissolution and In Vivo Pharmacokinetic Models.

In vitro and in vivo experimental models are frequently used to assess a new chemical entity's (NCE) biopharmaceutical performance risk for food effect (FE) in humans. Their ability to predict human FE hinges on replicating key features of clinical FE studies and building an in vitro-in vivo relationship (IVIVR). In this study, 22 compounds that span a wide range of physicochemical properties, Biopharmaceutics Classification System (BCS) classes, and food sensitivity were evaluated for biorelevant dissolution in fasted- and fed-state intestinal media and the dog fed/fasted-state pharmacokinetic model. Using the area under the curve (AUC) as a performance measure, the ratio of the fed-to-fasted AUC (FE ratio) was used to correlate each experimental model to FE ratio in humans. A linear correlation was observed for the in vitro dissolution-human IVIVR (R (2) = 0.66, % mean square error 20.7%). Similarly, the dog FE ratio correlated linearly with the FE ratio in humans (R (2) = 0.74, % mean square error 16.25%) for 15 compounds. Data points near the correlation line indicate dissolution-driven mechanism for food effect, while deviations from the correlation line shed light on unique mechanisms that can come into play such as GI physiology or unusual physicochemical properties. In summary, fed/fasted dissolution studies and dog PK studies show a reasonable correlation to human FE, hence are useful tools to flag high-risk NCEs entering clinical development. Combining kinetic dissolution, dog FE model and in silico modeling one can study FE mechanism and formulation strategies to mitigate the FE risk.

Assessing the risk of pH-dependent absorption for new molecular entities: a novel in vitro dissolution test, physicochemical analysis, and risk assessment strategy.

Weak base therapeutic agents can show reduced absorption or large pharmacokinetic variability when coadministered with pH-modifying agents, or in achlorhydria disease states, due to reduced dissolution rate and/or solubility at high gastric pH. This is often referred to as pH-effect. The goal of this study was to understand why some drugs exhibit a stronger pH-effect than others. To study this, an API-sparing, two-stage, in vitro microdissolution test was developed to generate drug dissolution, supersaturation, and precipitation kinetic data under conditions that mimic the dynamic pH changes in the gastrointestinal tract. In vitro dissolution was assessed for a chemically diverse set of compounds under high pH and low pH, analogous to elevated and normal gastric pH conditions observed in pH-modifier cotreated and untreated subjects, respectively. Represented as a ratio between the conditions, the in vitro pH-effect correlated linearly with clinical pH-effect based on the Cmax ratio and in a non-linear relationship based on AUC ratio. Additionally, several in silico approaches that use the in vitro dissolution data were found to be reasonably predictive of the clinical pH-effect. To explore the hypothesis that physicochemical properties are predictors of clinical pH-effect, statistical correlation analyses were conducted using linear sequential feature selection and partial least-squares regression. Physicochemical parameters did not show statistically significant linear correlations to clinical pH-effect for this data set, which highlights the complexity and poorly understood nature of the interplay between parameters. Finally, a strategy is proposed for implementation early in clinical development, to systematically assess the risk of clinical pH-effect for new molecular entities that integrates physicochemical analysis and in vitro, in vivo and in silico methods.

Compressive follower load influences cervical spine kinematics and kinetics during simulated head-first impact in an in vitro model.

Current understanding of the biomechanics of cervical spine injuries in head-first impact is based on decades of epidemiology, mathematical models, and in vitro experimental studies. Recent mathematical modeling suggests that muscle activation and muscle forces influence injury risk and mechanics in head-first impact. It is also known that muscle forces are central to the overall physiologic stability of the cervical spine. Despite this knowledge, the vast majority of in vitro head-first impact models do not incorporate musculature. We hypothesize that the simulation of the stabilizing mechanisms of musculature during head-first osteoligamentous cervical spine experiments will influence the resulting kinematics and injury mechanisms. Therefore, the objective of this study was to document differences in the kinematics, kinetics, and injuries of ex vivo osteoligamentous human cervical spine and surrogate head complexes that were instrumented with simulated musculature relative to specimens that were not instrumented with musculature. We simulated a head-first impact (3 m/s impact speed) using cervical spines and surrogate head specimens (n = 12). Six spines were instrumented with a follower load to simulate in vivo compressive muscle forces, while six were not. The principal finding was that the axial coupling of the cervical column between the head and the base of the cervical spine (T1) was increased in specimens with follower load. Increased axial coupling was indicated by a significantly reduced time between head impact and peak neck reaction force (p = 0.004) (and time to injury (p = 0.009)) in complexes with follower load relative to complexes without follower load. Kinematic reconstruction of vertebral motions indicated that all specimens experienced hyperextension and the spectrum of injuries in all specimens were consistent with a primary hyperextension injury mechanism. These preliminary results suggest that simulating follower load that may be similar to in vivo muscle forces results in significantly different impact kinetics than in similar biomechanical tests where musculature is not simulated.

Ex vivo measurement of lumbar intervertebral disc pressure using fibre-Bragg gratings.

Methods were developed to measure intervertebral disc pressure using optical fibre-Bragg gratings (FBGs). The FBG sensor was calibrated for hydrostatic pressure in a purpose-built apparatus and the average sensitivity was determined to be -5.7 +/- 0.085 pm/MPa (mean +/- SD). The average coefficient of determination (r(2)) for the calibration data was 0.99, and the average hysteresis of the sensor was 2.13% of full scale. The FBG was used to measure intradiscal pressure response to compressive load in five lumbar functional spine units. The pressure measured by the FBG sensor varied linearly with applied compressive load with coefficients of determination ranging from 0.84 to 0.97. The FBG sensor's sensitivity to compressive load ranged from 0.702 +/- 0.043 kPa/N (mean +/- SD) in a L1-L2 specimen, to 1.07 +/- 0.069 kPa/N in a L4-L5 specimen. These measurements agree with those of previous studies in lumbar spines. Two strain gauge pressure sensors were also used to measure intradiscal pressure response to compressive load. The measured pressure sensitivity to load ranged from 0.251 kPa/N (L4-L5) to 0.850 kPa/N (L2-L3). The average difference in pressure sensitivity to load between Sensors 1 and 2 was 12.9% of the value for Sensor 1, with a range from 1.1% to 20.4%, which suggests that disc pressure was not purely hydrostatic. This may have contributed to the difference between the responses of the FBG and strain gauge sensors.