ABSTRACT
BACKGROUND AND OBJECTIVES: A clinical trial was conducted to measure and analyse the pharmacokinetic parameters of a lipid formulation of risperidone, VAL401. The VAL401 formulation is designed to repurpose risperidone from an antipsychotic to an adenocarcinoma treatment, with the lipid formulation altering the cellular uptake of risperidone, thus enabling anticancer biology to be exhibited in preclinical testing. METHODS: This first human trial of VAL401 measured the concentrations of risperidone and its primary metabolite, 9-hydroxyrisperidone, in the blood of patients after treatment with a single 2-mg dose of VAL401. RESULTS: The trial provided information on differences in the pharmacokinetic profile of risperidone in VAL401 that may be caused by the formulation and/or the nature of the cancer patient population. VAL401 provided the following key pharmacokinetic parameters for the risperidone plasma concentration after a single 2-mg dose of VAL401, with results normalised to a dosage of 1 mg for comparison with literature values: Tmax, 2 h; Cmax, 8 ng/ml; half-life, 3.5 h; area under the plasma concentration-time curve from time zero to infinity (AUC0-∞), 58.2 ng h2/mL. CONCLUSIONS: Further comparisons of the pharmacokinetic parameters of risperidone and 9-hydroxyrisperidone in plasma of patients administered VAL401 and the corresponding parameters obtained from published data for conventionally formulated risperidone provide evidence for altered biological processing of VAL401 as compared to risperidone. The absolute values obtained provide support for future studies of VAL401 as a cancer treatment, as the Cmax demonstrates sufficient exposure to reach the concentrations seen during preclinical anticancer testing, yet the overall exposure to the active moiety supports the use of the safety and tolerability data from conventional risperidone during future clinical trials.
Subject(s)
Adenocarcinoma of Lung/metabolism , Antineoplastic Agents/pharmacokinetics , Carcinoma, Non-Small-Cell Lung/metabolism , Lipids/pharmacokinetics , Risperidone/pharmacokinetics , Adenocarcinoma of Lung/drug therapy , Adult , Aged , Antipsychotic Agents/pharmacokinetics , Area Under Curve , Carcinoma, Non-Small-Cell Lung/drug therapy , Chemistry, Pharmaceutical/methods , Female , Half-Life , Humans , Male , Middle Aged , Paliperidone Palmitate/pharmacokinetics , Therapeutic EquivalencyABSTRACT
We show that the use of multiple photochemistries is necessary to ensure diverse immobilisation of small molecules for binding of polypeptides using phage display and antibody libraries.
Subject(s)
Drug Design , Peptide Library , Peptides/metabolism , PhotochemistryABSTRACT
Drug development has moved along way forward from the days of with doctors peddling cauldrons of herbs and spices, however, the process can still miss opportunities for full exploitation of a drug's potential. Drug reprofiling provides a chance for an established or a forgotten drug to move into a new area of therapy, whether related to the known effects or in a completely new area. In an era of environmental awareness and spiraling costs for traditional drug development, a strategy to squeeze every benefit out of drugs with known safety, tolerability and pharmacological parameters must be a strategically sound desire. We explore examples of success in reprofiling, draw comparisons between techniques, and finally provide two examples from the Valirx plc development pipeline currently undergoing the process.
Subject(s)
Drug Repositioning , Intellectual Property , Off-Label UseABSTRACT
Drug reprofiling is emerging as an effective paradigm for discovery of cancer treatments. Herein, an antipsychotic drug is immobilised using the Magic Tag® chemical genomics tool and screened against a T7 bacteriophage displayed library of polypeptides from Drosophila melanogaster, as a whole genome model, to uncover an interaction with a section of 17-ß-HSD10, a proposed prostate cancer target. A computational study and enzyme inhibition assay with full length human 17-ß-HSD10 identifies risperidone as a drug reprofiling candidate. When formulated with rumenic acid, risperidone slows proliferation of PC3 prostate cancer cells in vitro and retards PC3 prostate cancer tumour growth in vivo in xenografts in mice, presenting an opportunity to reprofile risperidone as a cancer treatment.
Subject(s)
17-Hydroxysteroid Dehydrogenases/antagonists & inhibitors , 3-Hydroxyacyl CoA Dehydrogenases/antagonists & inhibitors , Adenocarcinoma/drug therapy , Antineoplastic Agents/pharmacology , Antipsychotic Agents/pharmacology , Drosophila melanogaster/drug effects , Drug Repositioning/methods , Enzyme Inhibitors/pharmacology , Genomics/methods , Prostatic Neoplasms/drug therapy , Risperidone/pharmacology , 17-Hydroxysteroid Dehydrogenases/chemistry , 17-Hydroxysteroid Dehydrogenases/genetics , 17-Hydroxysteroid Dehydrogenases/metabolism , 3-Hydroxyacyl CoA Dehydrogenases/chemistry , 3-Hydroxyacyl CoA Dehydrogenases/genetics , 3-Hydroxyacyl CoA Dehydrogenases/metabolism , Adenocarcinoma/enzymology , Adenocarcinoma/genetics , Animals , Antineoplastic Agents/chemistry , Antipsychotic Agents/chemistry , Bacteriophage T7/genetics , Cell Line, Tumor , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , Drosophila melanogaster/enzymology , Drosophila melanogaster/genetics , Drug Compounding , Enzyme Inhibitors/chemistry , Gene Library , Humans , Linoleic Acids, Conjugated/chemistry , Male , Mice, Nude , Molecular Docking Simulation , Molecular Targeted Therapy , Prostatic Neoplasms/enzymology , Prostatic Neoplasms/genetics , Protein Conformation , Risperidone/chemistry , Structure-Activity Relationship , Time Factors , Tumor Burden/drug effects , Xenograft Model Antitumor AssaysABSTRACT
Finding pleiomorphic targets for drugs allows new indications or warnings for treatment to be identified. As test of concept, we applied a new chemical genomics approach to uncover additional targets for the widely prescribed lipid-lowering pro-drug simvastatin. We used mRNA extracted from internal mammary artery from patients undergoing coronary artery surgery to prepare a viral cardiovascular protein library, using T7 bacteriophage. We then studied interactions of clones of the bacteriophage, each expressing a different cardiovascular polypeptide, with surface-bound simvastatin in 96-well plates. To maximise likelihood of identifying meaningful interactions between simvastatin and vascular peptides, we used a validated photo-immobilisation method to apply a series of different chemical linkers to bind simvastatin so as to present multiple orientations of its constituent components to potential targets. Three rounds of biopanning identified consistent interaction with the clone expressing part of the gene GJC3, which maps to Homo sapiens chromosome 7, and codes for gap junction gamma-3 protein, also known as connexin 30.2/31.3 (mouse connexin Cx29). Further analysis indicated the binding site to be for the N-terminal domain putatively 'regulating' connexin hemichannel and gap junction pores. Using immunohistochemistry we found connexin 30.2/31.3 to be present in samples of artery similar to those used to prepare the bacteriophage library. Surface plasmon resonance revealed that a 25 amino acid synthetic peptide representing the discovered N-terminus did not interact with simvastatin lactone, but did bind to the hydrolysed HMG CoA inhibitor, simvastatin acid. This interaction was also seen for fluvastatin. The gap junction blockers carbenoxolone and flufenamic acid also interacted with the same peptide providing insight into potential site of binding. These findings raise key questions about the functional significance of GJC3 transcripts in the vasculature and other tissues, and this connexin's role in therapeutic and adverse effects of statins in a range of disease states.
Subject(s)
Bacteriophage T7/genetics , Connexins/chemistry , Coronary Vessels/chemistry , Fatty Acids, Monounsaturated/chemistry , Hydroxymethylglutaryl-CoA Reductase Inhibitors/chemistry , Indoles/chemistry , Nerve Tissue Proteins/chemistry , Simvastatin/chemistry , Amino Acid Sequence , Biotransformation , Connexins/genetics , Coronary Vessels/surgery , Fluvastatin , Gene Expression , Humans , Models, Molecular , Molecular Sequence Data , Nerve Tissue Proteins/genetics , Peptide Library , Peptides/chemical synthesis , Peptides/chemistry , Pharmacogenetics , Photochemical Processes , Prodrugs/chemistry , Protein Binding , Protein Interaction Domains and Motifs , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Sequence Alignment , Simvastatin/analogs & derivativesABSTRACT
Magic Tag® immobilisation of bioactive molecules coupled with bacteriophage display, followed by an ELISA assay, provides a protocol that can probe interactions of drugs with putative products of alternative initiation of translation as exemplified by the binding of immobilised flecainide to protein products of genes linked to sudden cardiac death.
Subject(s)
Anti-Arrhythmia Agents/metabolism , Drosophila Proteins/genetics , Drosophila/genetics , Flecainide/metabolism , Genomics/methods , Reading Frames , Amino Acid Sequence , Animals , Bacteriophage T7/genetics , Base Sequence , Drosophila Proteins/chemistry , Drosophila Proteins/metabolism , Enzyme-Linked Immunosorbent Assay , Molecular Sequence Data , Peptide Library , Peptides/chemistry , Peptides/genetics , Peptides/metabolism , Protein BiosynthesisABSTRACT
We demonstrate the expected preference of an immobilised oligosaccharide Man(9)(GlcNAc)(2) upon a 96-well photochemical array, for its known receptor, the cell-surface lectin Dendritic Cell-Specific ICAM3 Grabbing Nonintegrin (DC-SIGN) when compared to immobilised competing monosaccharides.
Subject(s)
Cell Adhesion Molecules/metabolism , Lectins, C-Type/metabolism , Mannose/chemistry , Oligosaccharides/chemistry , Oligosaccharides/metabolism , Photochemistry/methods , Receptors, Cell Surface/metabolism , Carbohydrate Sequence , Humans , Molecular Sequence DataABSTRACT
Novel surfaces derivatized with tertiary amine oxides have been prepared and tested for their ability to resist nonspecific protein adsorption. The oxidation of tertiary amines supported on triazine units was carried out using mCPBA to give a format allowing conjugation of biologically active ligands alongside them. Adsorption to these surfaces was tested and compared to adsorption to a set of commercial and custom oligo-/poly(ethylene glycol) (OEG/PEG) supports by challenging them with a protein display library presented on bacteriophage lambda. The new class of amine oxide surfaces is found to compare favorably with the performance of the OEG/PEG supports in the prevention of nonspecific binding.