The combi-targetinG concept: intracellular fragmentation of the binary epidermal growth factor (EGFR)/DNA targeting "combi-triazene" SMA41.
We have designed a novel tumor targeting strategy that consists of designing molecules termed "combi-molecules" or TZ-I to be masked forms of multiple antitumor agents. One such molecule SMA41, the TZ-I prototype, has been shown to target the epidermal growth factor receptor (EGFR) and to degrade under physiological conditions to give SMA52 (I) (an inhibitor of EGFR) and methyldiazonium (TZ) (a DNA alkylating species). While the antiproliferative advantages of this novel binary targeting strategy have now been demonstrated, the exact subcellular localization of the degradation products released from SMA41 remained elusive. Here we exploited the fluorescence properties of SMA52 to study its release from SMA41 and its subcellular distribution. Further, using 14C-labeled SMA41, we determined the distribution of the methydiazonium within subcellular macromolecules (DNA, RNA, protein). The results showed that SMA41 degraded to SMA52 in the carcinoma of the vulva cell line A431 with a half-life of 11min. The latter compound was primarily distributed in the perinuclear region. At equimolar concentrations, higher levels of SMA52 were observed when released from SM41 than when the cells were directly exposed to SMA52, indicating that the combi-molecular approach may offer a transport advantage to the released bioactive species. Radioactivity associated with SMA41 3-[ 14C]-methyl group was distributed throughout DNA, RNA, and protein, the latter macromolecule being the most alkylated. The results suggest that SMA41 (TZ-I) may diffuse into the cells, break down into two species: SMA52 (I) concentrated in the perinuclear region and methyldiazonium (TZ) that diffuses in all intracellular organelles and unspecifically alkylates RNA, protein, and nuclear DNA.
Cancer Drug Research Laboratory, McGill University Health Center/Royal Victoria Hospital, M7.19, 687 Pine Ave. West, Montreal, Que., Canada H3A 1A1.
Stephanie L
SL
Matheson
Fouad
F
Brahimi
Bertrand J
BJ
Jean-Claude
D002455:Cell Division; D004247:DNA / Q000032:analysis*; D004815:Epidermal Growth Factor / Q000032:analysis*; D005434:Flow Cytometry / Q000379:methods; D005470:Fluorometry / Q000379:methods; D006801:Humans; D006868:Hydrolysis; D008856:Microscopy, Fluorescence / Q000379:methods; D011799:Quinazolines / Q000378:metabolism*; D012313:RNA / Q000032:analysis; D014407:Tumor Cells, Cultured; D014466:Ultraviolet Rays
D011799:Quinazolines; C421705:SMA-41; D004815:Epidermal Growth Factor; D012313:RNA; D004247:DNA