In vitro effect of chlorambucil on human glioma cell lines (SF767 and U87-MG), and human microvascular endothelial cell (HMVEC) and endothelial progenitor cells (ECFCs), in the context of plasma chlorambucil concentrations in tumor-bearing dogs

• Published in: PloS one 2018-09, Vol.13 (9), p.e0203517-e0203517
• Main Authors: Reese, Michael J, Knapp, Deborah W, Anderson, Kimberly M, Mund, Julie A, Case, Jamie, Jones, David R, Packer, Rebecca A
• Format: Article
• Language: English
• Subjects: Angiogenesis; Animals; Antineoplastic Agents, Alkylating - blood; Antineoplastic Agents, Alkylating - pharmacokinetics; Antineoplastic Agents, Alkylating - pharmacology; Apoptosis; Biology and Life Sciences; Biotechnology; Blood-brain barrier; Brain cancer; Cancer therapies; Cell Line, Tumor; Cell Proliferation - drug effects; Cell Survival - drug effects; Cells (biology); Cells, Cultured; Chemotherapy; Chlorambucil; Chlorambucil - blood; Chlorambucil - pharmacokinetics; Chlorambucil - pharmacology; Cytotoxicity; Dogs; Dose-Response Relationship, Drug; Drug dosages; Endothelial cells; Endothelial Cells - cytology; Endothelial Cells - drug effects; Endothelial Progenitor Cells - cytology; Endothelial Progenitor Cells - drug effects; Glioma; Glioma - blood; Glioma - blood supply; Glioma - metabolism; Glioma cells; Humans; Medical prognosis; Medicine and Health Sciences; Metabolic Clearance Rate; Microvasculature; Pharmacokinetics; Pharmacology; Progenitor cells; Radiation therapy; Research and Analysis Methods; Stem cells; Toxicity; Tumors; Veterinary colleges; Veterinary medicine
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0203517

The objective of this study was to investigate a possible mechanism of action of metronomic chlorambucil on glioma by studying the in vitro cytotoxicity and anti-angiogenic effects on glioma and endothelial cells, respectively. The in vitro LD50 and IC50 of chlorambucil were determined using human SF767 and U87-MG glioma cell lines, human microvascular endothelial cells (HMVECs) and human endothelial colony forming cells (ECFCs). Results were analyzed in the context of chlorambucil concentrations measured in the plasma of tumor-bearing dogs receiving 4 mg m-2 metronomic chlorambucil. The LD50 and IC50 of chlorambucil were 270 μM and 114 μM for SF767, and 390 μM and 96 μM for U87-MG, respectively. The IC50 of chlorambucil was 0.53 μM and 145 μM for the HMVECs and ECFCs, respectively. In pharmacokinetic studies, the mean plasma Cmax of chlorambucil was 0.06 μM. Results suggest that metronomic chlorambucil in dogs does not achieve plasma concentrations high enough to cause direct cytotoxic or growth inhibitory effects on either glioma or endothelial cells.