A PKPD Case Study: Achieving Clinically Relevant Exposures of AZD5991 in Oncology Mouse Models.

Capturing human equivalent drug exposures preclinically is a key challenge in the translational process. Motivated by the need to recapitulate the pharmacokinetic (PK) profile of the clinical stage Mcl-1 inhibitor AZD5991 in mice, we describe the methodology used to develop a refined mathematical model relating clinically relevant concentration profiles to efficacy. Administration routes were explored to achieve target exposures matching the clinical exposure of AZD5991. Intravenous infusion using vascular access button (VAB) technology was found to best reproduce clinical target exposures of AZD5991 in mice. Exposure-efficacy relationships were evaluated, demonstrating that dissimilar PK profiles result in differences in target engagement and efficacy outcomes. Thus, these data underscore the importance of accurately ascribing key PK metrics in the translational process to enable clinically meaningful predictions of efficacy.

A chronic bile duct and intravenous cannulation model in conscious rabbits for pharmacokinetic studies.

In the development of novel pharmaceutical compounds, pharmacokinetic parameters, such as the extent of biliary excretion, must be characterized. Pharmacokinetic studies in nonrodent species, typically dogs, are generally required for new drug approvals. However, in some cases, rabbits may be a more desirable model. We developed a surgical procedure for the intermittent or continuous collection of bile for long-term use. This surgery involves the removal of the gallbladder and cannulation of the proximal and distal aspects of the common bile duct using a 40 cm flexible surgical-grade cannula. The cannula loop is passed subcutaneously and exteriorized between the scapulae to divert bile flow. During use, the proximal cannula segment is attached to a collection container placed in a nylon torso jacket worn by the animal, and the distal segment of catheter is sealed with a stainless-steel adapter. An auricular catheter, secured by ligatures, is placed aseptically into the lateral ear vein and the tip is advanced to the cranial vena cava for serial blood collection. Daily infusions of a heparinized saline "lock" ensure patency. These procedures have been used in 18 rabbits for up to 1 month without clinical complications. Complete blood counts, biochemical profiles, body weights, and bile flow were monitored weekly and reflected normal enterohepatic circulation.

Early prediction of polymyxin-induced nephrotoxicity with next-generation urinary kidney injury biomarkers.

Despite six decades of clinical experience with the polymyxin class of antibiotics, their dose-limiting nephrotoxicity remains difficult to predict due to a paucity of sensitive biomarkers. Here, we evaluate the performance of standard of care and next-generation biomarkers of renal injury in the detection and monitoring of polymyxin-induced acute kidney injury in male Han Wistar rats using colistin (polymyxin E) and a polymyxin B (PMB) derivative with reduced nephrotoxicity, PMB nonapeptide (PMBN). This study provides the first histopathological and biomarker analysis of PMBN, an important test of the hypothesis that fatty acid modifications and charge reductions in polymyxins can reduce their nephrotoxicity. The results indicate that alterations in a panel of urinary kidney injury biomarkers can be used to monitor histopathological injury, with Kim-1 and α-GST emerging as the most sensitive biomarkers outperforming clinical standards of care, serum or plasma creatinine and blood urea nitrogen. To enable the prediction of polymyxin-induced nephrotoxicity, an in vitro cytotoxicity assay was employed using human proximal tubule epithelial cells (HK-2). Cytotoxicity data in these HK-2 cells correlated with the renal toxicity detected via safety biomarker data and histopathological evaluation, suggesting that in vitro and in vivo methods can be incorporated within a screening cascade to prioritize polymyxin class analogs with more favorable renal toxicity profiles.

Renal biomarker changes associated with hyaline droplet nephropathy in rats are time and potentially compound dependent.

Alpha 2u-globulin mediated hyaline droplet nephropathy (HDN) is a male rat specific lesion induced when a compound or metabolite binds to alpha 2u-globulin. The objective of this study was to investigate if the newer and more sensitive renal biomarkers would be altered with HDN as well as be able to distinguish between HDN and oxidative stress-induced kidney injury. Rats were dosed orally for 7 days to determine (1) if HDN (induced by 2-propanol or D-limonene) altered the newer renal biomarkers and not BUN or creatinine, (2) if renal biomarkers could distinguish between HDN and oxidative stress-induced kidney injury (induced by potassium bromate), (3) sensitivity of HDN-induced renal biomarker changes relative to D-limonene dose, and (4) reversibility of HDN and renal biomarkers, using vehicle or 300 mg/kg/day D-limonene with 7 days of dosing and necropsies scheduled over the period of Days 8-85. HDN-induced renal biomarker changes in male rats were potentially compound specific: (1) 2-propanol induced mild HDN without increased renal biomarkers, (2) potassium bromate induced moderate HDN with increased clusterin, and (3) D-limonene induced marked HDN with increased αGST, µGST and albumin. Administration of potassium bromate did not result in oxidative stress-induced kidney injury, based on histopathology and renal biomarkers creatinine and BUN. The compound D-limonene induced a dose dependent increase in HDN severity and renal biomarker changes without altering BUN, creatinine or NAG: (1) minimal induction of HDN and no altered biomarkers at 10 mg/kg/day, (2) mild induction of HDN with increased αGST and µGST at 50 mg/kg/day and (3) marked induction of HDN with increased αGST, µGST and albumin at 300 mg/kg/day. HDN induced by D-limonene was reversible, but with a variable renal biomarker pattern over time: Day 8 there was increased αGST, µGST and albumin; on Day 15 increased clusterin, albumin and Kim-1. In summary, HDN altered the newer and more sensitive renal biomarkers in a time and possibly compound dependent manner.

An acute rat in vivo screening model to predict compounds that alter blood glucose and/or insulin regulation.

INTRODUCTION: Drug-induced glucose dysregulation and insulin resistance have been associated with weight gain and potential induction and/or exacerbation of diabetes mellitus in the clinic suggesting they may be safety biomarkers when developing antipsychotics. Glucose and insulin have also been suggested as potential efficacy biomarkers for some oncology compounds. The objective of this study was to qualify a medium throughput rat in vivo acute Intravenous Glucose Tolerance Test (IVGTT) for predicting compounds that will induce altered blood glucose and/or insulin levels. METHODS: Acute and sub-chronic studies were performed to qualify an acute IVGTT model. Double cannulated male rats (Han-Wistar and Sprague-Dawley) were administered vehicle, olanzapine, aripiprazole or other compounds at t=-44min for acute studies and at time=-44min on the last day of dosing for sub-chronic studies, treated with dextrose (time=0min; i.v.) and blood collected using an automated Culex® system for glucose and insulin analysis (time=-45, -1, 2, 10, 15, 30, 45, 60, 75, 90, 120, 150 and 180min). RESULTS: Olanzapine significantly increased glucose and insulin area under the curve (AUC) values while aripiprazole AUC values were similar to control, in both acute and sub-chronic studies. All atypical antipsychotics evaluated were consistent with literature references of clinical weight gain. As efficacy biomarkers, insulin AUC but not glucose AUC values were increased with a compound known to have insulin growth factor-1 (IGF-1) activity, compared to control treatment. DISCUSSION: These studies qualified the medium throughput acute IVGTT model to more quickly screen compounds for 1) safety - the potential to elicit glucose dysregulation and/or insulin resistance and 2) efficacy - as a surrogate for compounds affecting the glucose and/or insulin regulatory pathways. These data demonstrate that the same in vivo rat model and assays can be used to predict both clinical safety and efficacy of compounds.

Optional inferior vena cava filter retrieval with retained thrombus: an in vitro model.

PURPOSE: Retrieval of an optional inferior vena cava (IVC) filter with retained thrombus may result in pulmonary emboli if the trapped thrombus is not removed along with the filter. An in vitro model was developed to determine the fate of trapped thrombus during filter removal. MATERIALS AND METHODS: An in vitro IVC flow model was created with 25-mm inner diameter tubing and a 50% glycerol/water solution. Three different optional filters-Recovery (Bard, Tempe, AZ), Günther-Tulip (Cook Inc., Bloomington IN), and OptEase (Cordis Endovascular/Johnson & Johnson, Warren, NJ)-were evaluated in the study. A known mass of mature thrombus (porcine, aged 1 wk) was trapped within the optional filters. The filters were then retrieved according to the manufacturers' protocol, and the mass of thrombus recovered with the filter was determined. For each filter, five iterations were performed with initial thrombus sizes less than 1 g (group A) and an additional five iterations with initial thrombus sizes greater than 1 g (group B). RESULTS: Thrombi from group A were statistically significantly smaller than those from group B (P < .0001). Retrieval of the Recovery filter resulted in an average of 25% (range, 0%-53%) and 4% (range, 0%-7%) of the clot being removed in group A and group B, respectively. Retrieval of the Günther-Tulip filter resulted in an average of 22% (group A) and 13% (group B) of the clot being removed. Retrieval of the OptEase filter resulted in an average of 43% (group A) and 0% (group B) of the clot being removed. CONCLUSIONS: In our in vitro model, we have established that the mass of thrombus retrieved with optional filters is only a fraction of the initial clot burden. Because of the risk of pulmonary emboli, care should be taken when IVC filters with large amounts of trapped thrombus are removed from patients.