Enhancing value of clinical pharmacodynamics in oncology drug development: An alliance between quantitative pharmacology and translational science.

Clinical pharmacodynamic evaluation is a key component of the "pharmacologic audit trail" in oncology drug development. We posit that its value can and should be greatly enhanced via application of a robust quantitative pharmacology framework informed by biologically mechanistic considerations. Herein, we illustrate examples of intersectional blindspots across the disciplines of quantitative pharmacology and translational science and offer a roadmap aimed at enhancing the caliber of clinical pharmacodynamic research in the development of oncology therapeutics.

Ultrafast laser functionalized rare phased gold-silicon/silicon oxide nanostructured hybrid biomaterials.

We introduce a hybrid nanostructured biomaterial that is a combination of rare phases of immiscible gold and silicon oxide, functionalized via ultrafast laser synthesis. For the first time, we show cancer controlling properties of rare phases of gold silicides, which include Au7Si, Au5Si, Au0.7Si2.3 and Au8Si2. Conventionally, pure forms of gold and silicon/silicon oxide are extensively employed in targeted therapy and drug delivery systems due to their unique properties. While silicon and silicon oxide nanoparticles have shown biocompatibility, gold nanoparticles show conflicting results based on their size and material properties. Several studies have shown that gold and silicon combinations produce cell controlling properties, however, these studies were not able to produce a homogenous combination of gold and silicon, owing to its immiscibility. A homogenous combination of gold and silicon may potentially enable properties that have not previously been reported. We describe rare phased gold-silicon oxide nanostructured hybrid biomaterials and its unique cancer controlling properties, owing to material properties, concentration, size and density. The gold-silicon oxide nanostructured hybrid is composed of individual gold-silicon oxide nanoparticles in various concentrations of gold and silicon, some nanoparticles possess a gold-core and silicon-shell like structure. The individual nanoparticles are bonded together forming a three dimensional nanostructured hybrid. The interaction of the nanostructured hybrids with cervical cancer cells showed a 96% reduction in 24h. This engineered nanostructured hybrid biomaterial presents significant potential due to the combination of immiscible gold and silicon oxide in varying phases and can potentially satiate the current vacuum in cancer therapy.

Ultrafast Laser Synthesized Nanostructures for Controlling Cell Proliferation.

The state-of-the-art in synthesis of nanostructured cell and contra-cell surfaces relies on techniques that utilize elaborate precursor chemicals, catalysts, or vacuum conditions, and any combination thereof. Two type s of nanostructures, sodium oxide (Na2O) nanotips and silicon oxide (SiO2) nanofibers, have been fabricated on soda-lime glass using ultrafast laser ablation. Control over nanotip width was demonstrated via laser dwell time and a new tip formation mechanism is proposed. The nanofibers generated in this work display a level of nanomorphology unseen in other fiber fabrication methods. The resulting fibers show striking morphological similarity to proteins that comprise the natural extra cellular matrix. The interaction of both nanostructures with NIH 3T3 fibroblasts was explored by incubating nanostructured glass with fibroblasts over periods of 12 hours, 1 day, or 1 week. The Na2O nanotip surfaces dissolved within a day yet appeared to induce apoptosis in cells while the SiO2 nanofibers degraded over time but influenced cells to display unique, healthy characteristics such as preferential adhesion to nanofibers and increased microvilli generation. These growth-positive and growth-negative surfaces for cells could find use in novel biological testing equipment.

Engineering functionalized multi-phased silicon/silicon oxide nano-biomaterials to passivate the aggressive proliferation of cancer.

Currently, the use of nano silicon in cancer therapy is limited as drug delivery vehicles and markers in imaging, not as manipulative/controlling agents. This is due to limited properties that native states of nano silicon and silicon oxides offers. We introduce nano-functionalized multi-phased silicon/silicon oxide biomaterials synthesized via ultrashort pulsed laser synthesis, with tunable properties that possess inherent cancer controlling properties that can passivate the progression of cancer. This nanostructured biomaterial is composed of individual functionalized nanoparticles made of a homogenous hybrid of multiple phases of silicon and silicon oxide in increasing concentration outwards from the core. The chemical properties of the proposed nanostructure such as number of phases, composition of phases and crystal orientation of each functionalized nanoparticle in the three dimensional nanostructure is defined based on precisely tuned ultrashort pulsed laser-material interaction mechanisms. The amorphous rich phased biomaterial shows a 30 fold (95%) reduction in number of cancer cells compared to bulk silicon in 48 hours. Further, the size of the cancer cells reduces by 76% from 24 to 48 hours. This method exposes untapped properties of combination of multiple phases of silicon oxides and its applications in cancer therapy.

Physiologically based and population PK modeling in optimizing drug development: A predict-learn-confirm analysis.

Physiologically based pharmacokinetic (PBPK) modeling and classical population pharmacokinetic (PK) model-based simulations are increasingly used to answer various drug development questions. In this study, we propose a methodology to optimize the development of drugs, primarily cleared by the kidney, using model-based approaches to determine the need for a dedicated renal impairment (RI) study. First, the impact of RI on drug exposure is simulated via PBPK modeling and then confirmed using classical population PK modeling of phase 2/3 data. This methodology was successfully evaluated and applied to an investigational agent, orteronel (nonsteroidal, reversible, selective 17,20-lyase inhibitor). A phase 1 RI study confirmed the accuracy of model-based predictions. Hence, for drugs eliminated primarily via renal clearance, this modeling approach can enable inclusion of patients with RI in phase 3 trials at appropriate doses, which may be an alternative to a dedicated RI study, or suggest that only a reduced-size study in severe RI may be sufficient.

Optimizing oncology therapeutics through quantitative translational and clinical pharmacology: challenges and opportunities.

Despite advances in biomedical research that have deepened our understanding of cancer hallmarks, resulting in the discovery and development of targeted therapies, the success rates of oncology drug development remain low. Opportunities remain for objective dose selection informed by exposure-response understanding to optimize the benefit-risk balance of novel therapies for cancer patients. This review article discusses the principles and applications of modeling and simulation approaches across the lifecycle of development of oncology therapeutics. Illustrative examples are used to convey the value gained from integration of quantitative clinical pharmacology strategies from the preclinical-translational phase through confirmatory clinical evaluation of efficacy and safety.

Evaluation of various static in vitro-in vivo extrapolation models for risk assessment of the CYP3A inhibition potential of an investigational drug.

Nine static models (seven basic and two mechanistic) and their respective cutoff values used for predicting cytochrome P450 3A (CYP3A) inhibition, as recommended by the US Food and Drug Administration and the European Medicines Agency, were evaluated using data from 119 clinical studies with orally administered midazolam as a substrate. Positive predictive error (PPE) and negative predictive error (NPE) rates were used to assess model performance, based on a cutoff of 1.25-fold change in midazolam area under the curve (AUC) by inhibitor. For reversible inhibition, basic models using total or unbound systemic inhibitor concentration [I] had high NPE rates (46-47%), whereas those using intestinal luminal ([I]gut) values had no NPE but a higher PPE. All basic models for time-dependent inhibition had no NPE and reasonable PPE rates (15-18%). Mechanistic static models that incorporate all interaction mechanisms and organ specific [I] values (enterocyte and hepatic inlet) provided a higher predictive precision, a slightly increased NPE, and a reasonable PPE. Various cutoffs for predicting the likelihood of CYP3A inhibition were evaluated for mechanistic models, and a cutoff of 1.25-fold change in midazolam AUC appears appropriate.

Mifamurtide in metastatic and recurrent osteosarcoma: a patient access study with pharmacokinetic, pharmacodynamic, and safety assessments.

PURPOSE: This non-randomized, patient-access protocol, assessed both safety and efficacy outcomes following liposomal muramyl-tripeptide-phosphatidylethanolamine (L-MTP-PE; mifamurtide) in patients with high-risk, recurrent and/or metastatic osteosarcoma. METHODS: Patients received mifamurtide 2 mg/m(2) intravenously twice-weekly ×12 weeks, then weekly ×24 weeks with and without chemotherapy. Serum concentration-time profiles were collected. Adverse events within 24 hours of drug administration were classified as infusion-related adverse events (IRAE); other AEs and overall survival (OS) were assessed. RESULTS: The study began therapy in January 2008; the last patient completed therapy in October 2012. Two hundred five patients were enrolled; median age was 16.0 years and 146/205 (71%) had active disease. Mifamurtide serum concentrations declined rapidly in the first 30 minutes post-infusion, then in a log-linear manner 2-6 hours post-dose; t1/2 was 2 hours. There were no readily apparent relationships between age and BSA-normalized clearance, half-life, or pharmacodynamic effects, supporting the dose of 2 mg/m(2) mifamurtide across the age range. Patients reported 3,679 IRAE after 7,482 mifamurtide infusions. These were very rarely grade 3 or 4 and most commonly included chills + fever or headache + fatigue symptom clusters. One- and 2-year OS was 71.7% and 45.9%. Patients with initial metastatic disease or progression approximated by within 9 months of diagnosis (N = 40) had similar 2-year OS (39.9%) as the entire cohort (45.9%) CONCLUSIONS: Mifamurtide had a manageable safety profile; PK/PD of mifamurtide in this patient access study was consistent with prior studies. Two-year OS was 45.9%. A randomized clinical trial would be required to definitively determine impact on patient outcomes.

The influence of laser-induced 3-D titania nanofibrous platforms on cell behavior.

The current challenge in tissue engineering is to design a platform that can provide appropriate topography and suitable surface chemistry to encourage desired cellular activities and to guide 3-D tissue regeneration. Compared with traditional cell culture materials, 3-D nanofibrous platforms offer a superior environment for promoting cell functions by mimicking the architecture of extracellular matrix (ECM). In this study, we present a technique to engineer freestanding 3-D titania nanofibrous structures on titanium substrates using femtosecond laser processing. The crystallinity, surface adhesion, and surface energy of the synthesized nanostructures are discussed. The effects of synthesized nanoarchitectures on the proliferation, morphology, and viability of MC3T3-E1 mouse osteoblast-like cells and NIH 3T3 mouse embryonic fibroblasts are investigated. The nanofibrous structures show high surface energy and hydrophilicity. The results from in vitro studies reveal that the titania nanofibrous architectures possess excellent biocompatibility and significantly enhances proliferation of both cell lines compared to untreated titanium specimens. Study of the cell morphology shows dynamic cell migration and attachment on the titania nanofibrous architecture. The bioactivity and biocompatibility of the engineered 3-D nanostructures suggest noticeable perspective for developing bio-functionalized scaffolds and implantable materials in regenerative medicine and clinical tissue engineering.

Femtosecond laser nanostructuring for femtosensitive DNA detection.

In this paper, a new concept to achieve improved probe-target recognition has been devised by introducing a novel class of DNA-functionalized three-dimensional (3D), stand-free, and nanostructured electrodes. The gold nanofibrous electrodes were created using MHZ ultrafast laser material processing in air at ambient conditions. The developed nanofibrous DNA biosensor was characterized by cyclic voltammetry with the use of ferrocyanide as an electrochemical redox indicator. Currently, electrochemical signal enhancement which is of great significance for improving the sensitivity in DNA detection remains a great challenge. Through, enhanced surface area-to-volume ratio and more efficient arrangement of probe molecules on nanofibrous electrodes, our newly developed electrode system overcomes some of the sensitivity challenges of the existing systems. This nanofiber-based system realizes femtomolar (fM) sensitivity toward complementary target DNA, and demonstrates a very wide dynamic range (from 1 fM to 1 nM).

Synthesis of bio-functionalized three-dimensional titania nanofibrous structures using femtosecond laser ablation.

The primary objective of current tissue regeneration research is to synthesize nano-based platforms that can induce guided, controlled, and rapid healing. Titanium nanotubes have been extensively considered as a new biomaterial for biosensors, implants, cell growth, tissue engineering, and drug delivery systems. However, due to their one-dimensional structure and chemical inertness, cell adhesion to nanotubes is poor. Therefore, further surface modification is required to enhance nanotube-cell interaction. Although there have been a considerable number of studies on growing titanium nanotubes, synthesizing a three-dimensional (3-D) nano-architecture which can act as a growth support platform for bone and stem cells has not been reported so far. Therefore, we present a novel technique to synthesize and grow 3-D titania interwoven nanofibrous structures on a titanium substrate using femtosecond laser irradiation under ambient conditions. This surface architecture incorporate the functions of 3-D nano-scaled topography and modified chemical properties to improve osseointegration while at the same time leaving space to deliver other functional agents. The results indicate that laser pulse repetition can control the density and pore size of engineered nanofibrous structures. In vitro experiments reveal that the titania nanofibrous architecture possesses excellent bioactivity and can induce rapid, uniform, and controllable bone-like apatite precipitation once immersed in simulated body fluid (SBF). This approach to synthesizing 3-D titania nanofibrous structures suggests considerable promise for the promotion of Ti interfacial properties to develop new functional biomaterials for various biomedical applications.

Single step self-enclosed fluidic channels via Two Photon Absorption (TPA) polymerization.

In this paper, we demonstrate a simple, fast and single-step method for fabricating self-enclosed fluidic channels via TPA. Pairs of parallel, polymerized ribs are linked by the subsequent polymerization with correctly predetermined offset between the ribs. The region, where the radicals are initiated but its concentration is below the threshold, we called it a sub-activated region. The subsequent polymerization is triggered by the overlap of the sub-activated regions of the two adjacent ribs. The dimensions of the self-enclosed channels depends on the offset between ribs, the scan speed as well as the laser parameters such as pulse energy, pulse width and repetition rate.

Maskless fabrication of nano-fluidic channels by two-photon absorption (TPA) polymerization of SU-8 on glass substrate.

We present a simple, fast, and repeatable method for fabricating nano-fluidic channels based on two-photon absorption (TPA) polymerization. Since our method does not require any mask, it is relatively cheaper and faster than other conventional lithography techniques. We illustrate that SU-8 has pronounced photoinitiation threshold behavior, which linearly decreases as the repetition rate increases. If the pulse energy and the repetition rate are controlled, channel width can be easily controlled. We report fluidic channels up to 110 nm in width, between polymerized parallel ribs, utilizing TPA and the photoinitiation threshold properties of SU-8. Finally, we also show that high repetition rate laser presents greater controllability in size of the polymerized region by varying fluence.

Synthesis of Nanoscale Tips Using Femtosecond Laser Radiation under Ambient Condition.

We report a unique growth of platelet-shaped nanoscale tips of transparent dielectric using femtosecond laser radiation at MHz pulse repetition rate with nitrogen background gas flow under ambient condition. The tips grew with sharp nanoscale apex while their base and lengths are of the order of few hundred nanometers. In the absence of nitrogen, the irradiation leads to nanofibrous structure formation. The collision between the nitrogen gas atoms and the vapor species slows down plume expansion and lead to an increase of nanoparticles size. This prevents the fibrous structure formation and provides appropriate condition for nanoscale tips growth.

Synthesis of Glass Nanofibers Using Femtosecond Laser Radiation Under Ambient Condition.

We report the unique growth of nanofibers in silica and borosilicate glass using femtosecond laser radiation at 8 MHz repetition rate and a pulse width of 214 fs in air at atmospheric pressure. The nanofibers are grown perpendicular to the substrate surface from the molten material in laser-drilled microvias where they intertwine and bundle up above the surface. The fibers are few tens of nanometers in thickness and up to several millimeters in length. Further, it is found that at some places nanoparticles are attached to the fiber surface along its length. Nanofiber growth is explained by the process of nanojets formed in the molten liquid due to pressure gradient induced from the laser pulses and subsequently drawn into fibers by the intense plasma pressure. The attachment of nanoparticles is due to the condensation of vapor in the plasma.

Mechanism-based inactivation of human cytochrome P450 enzymes: strategies for diagnosis and drug-drug interaction risk assessment.

Among drugs that cause pharmacokinetic drug-drug interactions, mechanism-based inactivators of cytochrome P450 represent several of those agents that cause interactions of the greatest magnitude. In vitro inactivation kinetic data can be used to predict the potential for new drugs to cause drug interactions in the clinic. However, several factors exist, each with its own uncertainty, that must be taken into account in order to predict the magnitude of interactions reliably. These include aspects of in vitro experimental design, an understanding of relevant in vivo concentrations of the inactivator, and the extent to which the inactivated enzyme is involved in the clearance of the affected drug. Additionally, the rate of enzyme degradation in vivo is also an important factor that needs to be considered in the prediction of the drug interaction magnitudes. To address mechanism-based inactivation for new drugs, various in vitro experimental approaches have been employed. The selection of approaches for in vitro kinetic characterization of inactivation as well as in vitro-in vivo extrapolation should be guided by the purpose of the exercise and the stage of drug discovery and development, with an increase in the level of sophistication throughout the research and development process.

Human drug metabolism and the cytochromes P450: application and relevance of in vitro models.

The cytochromes P450 (CYPs) constitute a superfamily of hemoprotein enzymes that are responsible for the biotransformation of numerous xenobiotics, including therapeutic agents. Studies of the biochemical and enzymatic properties of these enzymes and their molecular genetics and regulation of gene expression and activity have greatly enhanced our understanding of several aspects of clinical pharmacology such as pharmacokinetic variability, drug toxicity, and drug interactions. This review evaluates the major human hepatic drug-metabolizing CYP enzymes and their clinically relevant substrates, inhibitors, and inducers. Also discussed are the molecular bases and clinical implications of genetic polymorphisms that affect the CYPs. Much of the information on the specificity of substrates and inhibitors of the CYP enzymes is derived from in vitro studies using human liver microsomes and heterologously expressed CYP enzymes. These methods are discussed, and guidelines are provided for designing enzyme kinetic and reaction phenotyping studies using multiple approaches. The strengths, weaknesses, and discrepancies among the different approaches are considered using representative examples. The mathematical models used in predicting the pharmacokinetic clearance of a drug from in vitro estimates of intrinsic clearance and the principles of quantitative in vitro-in vivo scaling of metabolic drug interactions are also discussed.

Relative contribution of CYP3A to amitriptyline clearance in humans: in vitro and in vivo studies.

The relative contribution of cytochrome P450 3A (CYP3A) to the oral clearance of amitriptyline in humans has been assessed using a combination of in vitro approaches together with a clinical pharmacokinetic interaction study using the CYP3A-selective inhibitor ketoconazole. Lymphoblast-expressed CYPs were used to study amitriptyline N-demethylation and E-10 hydroxylation in vitro. The relative activity factor (RAF) approach was used to predict the relative contribution of each CYP isoform to the net hepatic intrinsic clearance (sum of N-demethylation and E-10 hydroxylation). Assuming no extrahepatic metabolism, the model-predicted contribution of CYP3A to net intrinsic clearance should equal the fractional decrement in apparent oral clearance of amitriptyline upon complete inhibition of the enzyme. This hypothesis was tested in a clinical study of amitriptyline (50 mg, p.o.) with ketoconazole (three 200 mg doses spaced 12 hours apart) in 8 healthy volunteers. The RAF approach predicted CYP2C19 to be the dominant contributor (34%), with a mean 21% contribution of CYP3A (range: 8%-42% in a panel of 12 human livers). The mean apparent oral clearance of amitriptyline in 8 human volunteers was decreased from 2791 ml/min in the control condition to 2069 ml/min with ketoconazole. The average 21% decrement (range: 2%-40%) was identical to the mean value predicted in vitro using the RAF approach. The central nervous system (CNS) sedative effects of amitriptyline were slightly greater when ketoconazole was coadministered, but the differences were not statistically significant. In conclusion, CYP3A plays a relatively minor role in amitriptyline clearance in vivo, which is consistent with in vitro predictions using the RAF approach.

Pharmacokinetics and pharmacodynamics of vecuronium in children receiving phenytoin or carbamazepine for chronic anticonvulsant therapy.

The pharmacokinetics and time course of action of vecuronium in normal children and children receiving anticonvulsant drugs for prolonged periods were characterized. A bolus dose of vecuronium 0.15 mg kg(-1) was administered i.v. to 10 non-epileptic children and to 10 children on phenytoin and 10 children on carbamazepine, who were matched for age and weight. Plasma concentrations of vecuronium, 3-OH desacetylvecuronium (the primary metabolite of vecuronium) and alpha1-acid glycoprotein (AAG) were determined. Pharmacokinetic variables were derived from plasma samples collected before and after administration of vecuronium. Neuromuscular transmission was monitored by evoked compound electromyography. Recovery of the first twitch of the train-of-four (T1/T0) and the recovery index (RI), the time for 25-75% recovery of T1/T0, were determined. The elimination half-life of vecuronium was significantly reduced in both anticonvulsant groups compared with control [control 48.2 (SD 40.3), phenytoin 23.5 (13.1), carbamazepine 18.4 (16.6) min, P<0.05]. Vecuronium clearance was increased in both anticonvulsant groups [control 9.0 (3.6), phenytoin 15.1 (8.9), carbamazepine 18.8 (13.1) ml kg(-1) min(-1), 0.05