The Penrose platform is a rational, chemistry-driven model exploiting the bio-redox states of malignant cells.
The model involves multiple scientific divisions: medicinal chemistry, analytical chemistry, protein chemistry, cell & molecular biology, and in vivo studies. Based on this model several new molecular entities have been designed and currently are being tested in high throughput screening on various cancers. It is a platform with potential broad application across multiple tumor types – a universal chemistry driven anticancer platform.
Our Scientific Divisions
Our Medicinal Chemistry Division designs, chemically synthesizes and develops anti-cancer and other bio-active small molecules and agents, utilizing understanding of structure-activity relationships (SAR) in the early drug discovery phase.
Our Analytical Chemistry Division utilizes classical wet chemical methods and modern instrumental methods to separate, identify, and quantify analytes of interest, from synthetic intermediates to metabolites, in a variety of sample matrices such as cancer cells, urine, tissues, and more.
Our Process Chemistry Division develops and optimizes synthetic routes and pilot plant procedures to manufacture safe, cost/labor efficient, green, and reproducible compounds for the drug development phase.
Our Cell Biology Division focuses on determining how cells function in response to our drugs and the significance of those functions in translation to drug discovery. Our scientists study the mechanisms that control the essential aspects of cell behavior, from the single cell level to the multicellular level. To understand the changes that occur inside of a cell during the course of its normal lifecycle as well as the diseased state, we utilize an interdisciplinary approach. We use bioassays, an important part of the FDA drug approval process, to determine the potency of our drugs on a variety of cell lines. We then compare the biological response related to its mode of action with that of a control preparation. Our scientists specialize in assay development and validation, and have expertise in cell viability and toxicity assays, cell proliferation assays for apoptosis and necrosis, cell signaling assays, and a variety of immunoassays including ELISA and Flow Cytometry.
We use flow cytometry to detect and measure changes in cellular processes including mitochondrial dynamics, cell proliferation, and cell death upon treatment of our anti-cancer agents. Our facility is equipped with a Cytek™ Aurora flow cytometer; with three lasers, two scattering channels, and up to 48 fluorescence channels, the Aurora system provides unprecedented performance for simple and high-complexity applications. Using enzyme activity and kinetics assays, we assess functional inhibition of critical metabolic enzymes by our anti-cancer agents, in both cell-based and purified enzyme systems. In addition, we will use chemical crosslinking followed by pulldown and mass spectrometry to identify protein targets of our anti-cancer agents.
Molecular Biology and Biochemistry
Our Molecular Biology and Biochemistry Division studies the metabolism and mechanism of the diseased state cells for the development of drugs; fundamentals in molecular biology and biochemistry hold key positions acting as interface between, chemistry, structural biology and cell biology. We are an innovative and creative group of people who are interested in complex problem solving to understand life at the organismal, macromolecular, and molecular levels. Our scientists devise strategies to use molecular and biochemical approaches to address questions in cancer biology.
Our interest in elucidating the mode of action of our drugs at every biological level invariably includes gene expression analysis as a fundamental piece in our puzzle. To do so we use polymerase chain reaction (PCR) techniques; PCR is a sensitive method broadly used in molecular biology to exponentially amplify specific DNA sequences. Penrose counts with the thermocycler ProFlex™ 3 x 32-well PCR System from Applied Biosystems™ Thermo Fisher Scientific to run PCR analysis. To further comprehend how our treatment affects gene expression in cancer cells, in real time, we use quantitative polymerase chain reaction (qPCR) analysis to precisely measure changes in mRNA levels. For that application, Penrose is equipped with a high-end qPCR system, the Applied Biosystems™ QuantStudio™ 6 Flex Real-Time PCR System and its intuitive and powerful QuantStudio™ Real-Time PCR Software, from Thermo Fisher Scientific.
To study the global changes in protein expression in cell and tissue extracts we use a variety of methods, from more traditional methods for detection and characterization of protein expression to fully automated systems; we are equipped for accurate and efficient studies. To analyze the effects of our therapies in biological systems we utilize a relatively new capillary electrophoresis based western blot technology, the Simple Western WES with compass software from Protein Simple, which allows for automation to provide rapid quantitative results.
As we are exploring the genes, proteins, and metabolites involved in the drug metabolism process, we are combining them with bioinformatics tools to better address our questions with the –omics approach: genomics, proteomics and metabolomics. This comprehensive perspective addresses target identification and metabolic network understanding, which is essential in generating future discoveries.
Cellular Imaging Division
Our Cellular Imaging Division focuses on providing visual clarity to the bio-chemical and bio-physical mechanisms undergone on the cell and tissue level in response to our drug administration.
The clinical samples generated by the multitude of divisions within Penrose are imaged in-house. We have a Nikon Eclipse Ci-L upright microscope with a Nikon DS-Fi3 Color Digital Camera System, alongside NIS Elements BR Software. This microscope gives superb images over the entire magnification range; we look at Hematoxylin and Eosin stains along with other histological staining. Our scientists for their wide range of studies use the facility for imaging subcellular structures in live and fixed cells. Immunohistochemistry to image discrete components in tissue samples are performed for extensive protein diagnosis studies.
Fluorescence imaging has advanced our knowledge of cellular biology and diseases at the molecular level. Our group is using the powerful tool of fluorescence imaging to explore a broad range of experimental observations; this includes studies involving location and dynamics of molecules, helping to show how our drugs localize and interact with cellular structures. With the help of fluorescent dyes and proteins as labels for molecular structures and processes, we access protein expression and gene expression in cells and tissues. Our imaging experiments involves organelle staining, immunocytochemistry, reporter gene assays etc., to support the basic science research in our organization. There has been rapid advancement in the field of optical imaging providing for higher resolution; this allows us to capture the finer details of biological processes occurring inside a cell or an organelle. Penrose is also building an imaging core with epifluorescence and confocal microscopes, with enhanced optical resolution, to understand the science through visual depiction of the biological processes involved in our mechanism.
“A picture is worth a thousand words” and a picture with a temporal dimension speaks even more. The dynamic changes inside a cell cannot be observed in a snapshot; we use live-cell imaging and analysis for observing cell structures and processes in real-time, and over a period of time. Furthermore, cellular integrity, protein trafficking, signal transduction, and enzyme activity can all be monitored. We are able to study the dynamic changes in the cell environment in response to the administration of drugs, including how molecules translocate and interact within the cell after the external stimuli. We use IncuCyte S3 Live-cell Analysis system for real time analysis of cell health and to acquire kinetic data. The instrument maintains the cells in a healthy state for extended periods of time. Ranging from proliferation, cytotoxicity, and spheroid assays, we conduct a number of live-cell assays for comprehension of the mechanism behind our therapy.
In Vivo Xenografts
Our Pharmacology Division uses cancer cell lines originally derived from patient tumors and grown as xenografts in immunocompromised mice models. Experiments within these models have played a significant role in the late-stage pre-clinical development of targeted anticancer therapies. We evaluate the ability of anticancer agents to reduce tumor growth in preclinical xenograft models to provide the basis for compound optimization. Pre-clinical in vivo xenograft models have been shown to be reasonably well-correlated with human clinical outcomes. At present we are in the process of testing various tumor models with different dose regimens and dosing schedules, helping us provide maximum efficacy and minimum toxicity.
We are also utilizing Patient-Derived Tumor Xenografts (PDX) models because they have the advantage of maintaining the molecular and histological heterogeneity of the original tumor. Moreover, they have been shown to be superior at predicting drug response compared to standard cell culture xenograft models. Using both cell line-based and patient-derived xenografts models for investigating cancer initiation, progression, and treatment response is critical as we optimize our treatment.
Pharmacokinetics and Pharmacodynamics
Pharmacokinetics/Pharmacodynamics plays an important role in drug development, as they can provide a basis for predicting drug effects in humans through pre-clinical in vivo experimental data. PK and PD will help us to identify the key properties of our drugs, and will allows us to characterize and predict the time-course effects of drugs under physiological and pathological conditions. Recent advances in systems biology and systems pharmacology have provided new information on how drugs affect the human body as a single complex biological system. We strongly believe this will give rise to new opportunities in drug combinations, which can only be developed rationally through the appropriate application of dynamic systems-based PK/PD models. The relationships between pharmacokinetics and pharmacodynamics observed in pre-clinical models support efficient dose selection, which is pivotal for successful clinical trials.
High-Throughput Screening Division
High-Throughput Drug Screening
Our High-Throughput Screening Division is designing bio-active molecules and anti-cancer drugs to create a library of active compounds. High-Throughput Screening (HTS) is a popular approach used in drug discovery for target validation. We measure the effects of our compounds through biological assays; the methods require the use of multiple colorimetric assays, along with imaging and computation to increase the scale and speed of performing the screens. Currently we use cell-viability assays to broadly identify the compounds that kill cancer cells, and their effect cell growth. We have additionally developed various end-point biochemical assays and image-based assays to accelerate the search for potential new drugs from our compound library. We combine imaging-based 3D models to our screening studies in order to generate biologically relevant data, and to increase the likelihood of success in human studies. Emerging assay technologies using time-resolved fluorescence, functional whole cell assays, and high-content assays, coupled with analytical techniques such as LC-MS/MS to integrate with automated HTS, we are able to support basic and translational biomedical research of our company. Our HTS Division identifies potential hits within our library of compounds, and pursues active compounds to elucidate their pharmacodynamics and pharmacokinetic properties; these further become leads for medicinal chemistry optimization.