Physicochemical characterization of substituted chromeno[4,3- b][1,5]benzodiazepine stereoisomers designed as cell membrane active antitumor agents

• Published in: Biophysical chemistry 1990, Vol.35 (2), p.271-285
• Main Authors: Werner, Walter, Baumgart, Joachim, Burckhardt, Günter, Fleck, Werner F., Geller, Klaus, Gutsche, Walter, Hanschmann, Helmut, Messerschmidt, Albrecht, Römer, Wolfgang, Tresselt, Dieter, Löber, Günter
• Format: Article
• Language: English
• Subjects: benzodiazepines; cell membranes; Chromeno[4,3- b][1,5]benzodiazepine stereoisomer, substituted; Fluorescence spectroscopy; NMR; Spectrophotometry; X-ray diffraction
• ISSN: 0301-4622; 1873-4200
• DOI: 10.1016/0301-4622(90)80015-Y

As an alternative to naturally occurring pyrrolo[2,1- c][1,4]benzodiazepines (e.g., antramycin) which possess properties of DNA alkylation, we have designed several antileukemic chromeno[4,3- b][1,5]benzodiazepine derivatives with potential activity toward leukemia cell membranes and the cyclic nucleotide system. The cis and trans diastereoisomers were characterized by NMR. The absolute configurations of the enantiomers were established by X-ray diffraction and circular dichroism (CD) measurements. By means of absorption spectroscopy and determinations of fluorescence and fluorescence decay, it was found that the cancerostatically active compound (+)(6 aR,13 aS)-3,4-dimethoxy-10,11-dimethyl-6, 6 a,7,8,13,13 a-hexahydrochromeno[4,3- b][1,5]benzodiazepine (ZIMET 54/79) and its biologically inactive (−) enantiomer (ZIMET 55/79) interact with liposomal membranes. At pH values of 6.0 and 7.3, the long-wave absorption bands of these agents showed weak bathochromic and hypochromic effects upon addition of neutral, and positively and negatively charged phosphatidylcholine and phosphatidylcholine/cholesterol liposomes. Such spectral changes are interpreted as resulting from the binding of both agents to phospholipid bilayers. Steady-state determinations using the membrane probe 1-anilino-8-naphthalenesulfonic acid (1,8-ANS) led to the observation of a small decrease in fluorescence intensity in the presence of either agent. Time-resolved measurements demonstrate that the mechanism of action of the agents occurs mainly through the partial displacement of probe molecules from regions of hydrophobic binding to areas of greater solvent accessibility. No significant differences in binding between the cancerostatically active and inactive enantiomers with liposomes (achiral systems) were detectable on the basis of spectrophotometric and fluorescence determinations. Cell membrane bound adenylate cyclase is stimulated by ZIMET 54/79, resulting in an increase of 103% in the level of cAMP in mouse L1210 leukemia cells. On examination of structure-activity relationships, it was found that the biological activity (leukemia L1210, P388, Lewis lung carcinoma, melanoma B16, increase in cAMP) is correlated with the particular configuration (6 aR,13 aS) and type of substituent at positions 3 and 4 of the benzo ring in the case of alkoxy groups and positions 10 and 11 for methyl groups. No activity was detected toward DNA/RNA using microbial test systems.