F. Takusagawa et al., PHYSICAL AND BIOLOGICAL CHARACTERISTICS OF THE ANTITUMOR DRUG ACTINOMYCIN-D ANALOGS DERIVATIZED AT N-METHYL-L-VALINE RESIDUES, Biochemistry, 35(40), 1996, pp. 13240-13249
The crystal structure of the DNA-actinomycin D (AMD) complex and a sim
ple molecular modeling study indicated that AMD analogues derivatized
at N-methyl-L-valine residues (fifth amino acid residue in the cyclic
depsipeptide of AMD) could bind to DNA as strongly as the parent AMD.
The analogues in which N-methyl-L-valine residues were replaced with L
- and D-forms of N-methylvalines, N-methylthreonines, N-methylphenylal
anines, N-methyltyrosines, and N-methyl-O-methyltyrosines have been to
tally synthesized. The characteristics of binding of the analogues to
various DNAs including DNA-1 [d(TATATATGCATATATA)], DNA-2 [d(TATATACGC
GTATATA)], DNA-3 [d(ATATATAGCTATATAT)], and DNA-4 [d(ATATATGGCCATATAT)
] have been examined by using visible absorption spectrum methods. The
association constants calculated from the absorption spectra indicate
that the modifications of the N-methyl-L-valine residues in the AMD m
olecule do affect the DNA binding characteristics of the analogues. Th
e L-aromatic analogues bind slightly better than the L-aliphatic analo
gues;except for binding to DNA-1 (-TGCA-), whereas the D-aliphatic ana
logues bind consistently better than the D-aromatic analogues. In the
L-form analogues, the L-Tyr analogue has the highest overall associati
on constant, whereas the D-Val analogue has the highest association co
nstant among the D-form analogues. In spite of substitution of bulky a
romatic groups, the D-aromatic analogues bind to the DNA-I quite well.
However, D-aromatic analogues have significantly reduced their bindin
g capacities to the other DNAs, indicating that the substitution of th
e D-aromatic residues creates a unique four-base sequence preference (
-TGCA-). The RNA polymerase inhibitory activities of the AMD analogues
in vivo have been examined using human cells (HeLa). All AMD analogue
s except for the L-Thr analogues severely inhibit RNA synthesis at rel
atively low drug concentrations. The D-Val, L-OMT, L-Phe, and D-Phe an
alogues inhibit RNA synthesis more strongly than the natural antibioti
c (AMD itself).