The interaction of a new anti‐tumour drug, KAR‐2 with calmodulin

• Published in: British journal of pharmacology Vol. 121; no. 5; pp. 955 - 962
• Main Authors: Orosz, F, Vértessy, B G, Salerno, C, Crifo, C, Capuozzo, E, Ovádi, J
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.07.1997; Nature Publishing
• Subjects: Animals; Antineoplastic agents; Antineoplastic Agents, Phytogenic - metabolism; Antineoplastic Agents, Phytogenic - pharmacology; Biological and medical sciences; bis‐indols; Calcium - metabolism; Calmodulin - antagonists & inhibitors; Calmodulin - metabolism; calmodulin antagonism; Cattle; Chemotherapy; Circular Dichroism; Enzyme-Linked Immunosorbent Assay; Humans; Immunoglobulin G; Isoenzymes - antagonists & inhibitors; Isoenzymes - metabolism; Medical sciences; Neutrophils - drug effects; Neutrophils - metabolism; Pharmacology. Drug treatments; Phosphofructokinase-1 - antagonists & inhibitors; Protein Binding; Spectrometry, Fluorescence; Spectrophotometry, Ultraviolet; Trifluoperazine - pharmacology; vinblastine; Vinblastine - analogs & derivatives; Vinblastine - metabolism; Vinblastine - pharmacology; Vinca alkaloids; vincristine; Antineoplastic agent; Human; Vinblastine; Calcium; Electrophysiology; Ionic current; In vitro; Alkaloid; Blood cell; Biological fixation; Structure activity relation; Neutrophil; Indole derivatives; Mechanism of action; Antimitotic; Calmodulin; Comparative study
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1038/sj.bjp.0701190

KAR‐2 (3′′‐(β‐chloroethyl)‐2′′,4′′‐dioxo‐3,5′′‐spiro‐oxazolidino‐4‐deacetoxy‐vinblastine) is a semisynthetic bis‐indol derivative, with high anti‐microtubular and anti‐tumour activities but with low toxicity. KAR‐2, in contrast to other biologically active bis‐indols (e.g. vinblastine), did not show anti‐calmodulin activity in vitro (enzyme kinetic, fluorescence anisotropy and immunological tests). Direct binding studies (fluorescence resonance energy transfer, circular dichroism) provided evidence for the binding of KAR‐2 to calmodulin. The binding affinity of KAR‐2 to calmodulin (dissociation constant was about 5 μM) in the presence of Ca2+ was comparable to that of vinblastine. KAR‐2 was able to interact with apo‐calmodulin as well; in the absence of Ca2+ the binding was of cooperative nature. The effect of drugs on Ca2+ homeostasis in human neutrophil cells was investigated by means of a specific fluorescent probe. Trifluoperazine extensively inhibited the elevation of intracellular Ca2+ level, vinblastine did not appreciably affect it, KAR‐2 stimulated the Ca2+ influx and after a transient enhancement the Ca2+ concentration reached a new steady‐state level. Comparison of the data obtained with KAR‐2 and bis‐indols used in chemotherapy suggests that the lack of anti‐calmodulin potency resides on the spiro‐oxazolidino portion of KAR‐2. This character of KAR‐2 manifested itself in various systems and might result in its low in vivo toxicity, established in an anti‐tumour test.