Experiments and Techniques

Natural Products Isolation and Characterization

  • Identity: Establish an HPLC fingerprint of dietary supplement grade material
  • Quantification: Standardize a plant/extract to 1 or more phytochemicals
  • Isolation: Separate individual phytochemicals from plants at >95% purity

Cell Free Assays

  • Surface Plasmon Resonance – Identify direct binders to specific drug targets
  • Nuclear receptor ligand binding – Identify small molecules that directly interact with the androgen receptor, estrogen receptor and additional nuclear receptors.
  • HMG-CoA reductase – Screen for director inhibitors that can disrupt cholesterol synthesis
  • P450 microsomes – Identify inhibitors of individual P450 enzymes (1A2, 2C9, 2C19, 2D6 and 3A4.

Cell Culture

  • Cell lines – Prostate (LNCaP, RWPE-1, 22Rv1, PC3, VCaP), Breast (MCF-7, MDA-MB-231), Colon (Caco-2, HT-29, HCT-116, SW480)
  • Assays – MTT, BrdU cell proliferation, Western Blot, qPCR, immunoflourescence microscopy, ELISA, siRNA

Animal Models

  • Pharmacokinetics –Single dose and multiple dose pharmacokinetics in mice. Study agent is administered orally and by intraperitoneal injection with blood and tissue collection.
  • Xenograft – Perform cell transplantation into athymic nude mice and administer study agents via diet, oral gavage, and intraperitoneal administration.
  • DSS Colitis – Administer dextran sodium sulfate to induce colitis and administer study agents. At completion can determine the disease activity index and evaluate for local effects in the gastrointestinal tract.

Pharmacokinetic Assays

  • LC-MS/MS assay development – Capable of detecting study agents in neat samples, blood, and tissue. This includes ion fragmentation, plasma extraction efficiency, standard curves, intra- and inter-day variability, and stability studies.
  • Developed LC-MS/MS Assays – Methods developed for resveratrol, green tea catechins (EGCG, EGC, EC, ECG), rosemary diterpenes (carnosic acid, carnosol), mangosteen xanthones (alpha-mangostin, gartanin), melphalan, clomiphene, letrozole. In addition, we have also developed assays for ~20 novel compounds each year for the UIC Drug Discovery Center.
  • Targeted Mass Spectometry (MRM) – Preliminary identification of metabolites derived from the study agent.
  • Microsome stability – Evaluation of study agent in liver or gastrointestinal microsomes to determine extent of study agent metabolism
  • Human clinical trials – Experience quantifying and determining the pharmacokinetic parameters of study agents in human volunteers.