3 research outputs found
Computational Studies Support the Role of the C7-Sibirosamine Sugar of the Pyrrolobenzodiazepine (PBD) Sibiromycin in Transcription Factor Inhibition
The
pyrroloÂ[2,1-<i>c</i>]Â[1,4]Âbenzodiazepines (PBDs)
are a group of sequence-selective, DNA minor-groove binding agents
that covalently attach to guanine residues. Originally derived from <i>Streptomyces</i> species, a number of naturally occurring PBD
monomers exist with varying A-Ring and C2-substituents. One such agent,
sibiromycin, is unusual in having a glycosyl residue (sibirosamine)
at its A-Ring C7-position. It is the most cytotoxic member of the
naturally occurring PBD family and has the highest DNA-binding affinity.
Recently, the analogue 9-deoxysibiromyin was produced biosynthetically
by Yonemoto and co-workers. Differing
only in the loss of the A-Ring C9-hydroxyl group, it was reported
to have a significantly higher DNA-binding affinity than sibiromycin
based on DNA thermal denaturation studies, although these data have
since been retracted. As deletion of the
C9-OH moiety, which points toward the DNA minor groove floor, might
intuitively be expected to reduce DNA-binding affinity through the
loss of hydrogen bonding, we carried out molecular dynamics simulations
on the interaction of both molecules with DNA over a 10 ns time-course
in explicit solvent. Our results suggest that the two molecules may
differ in their sequence-selectivity and that 9-deoxysibiromycin should
have a lower binding affinity for certain sequences of DNA compared
to sibiromycin. Our molecular dynamics results indicate that the C7-sibirosamine
sugar does not form hydrogen bonding interactions with groups in the
DNA minor-groove wall as previously reported, but instead points orthogonally
out from the minor groove where it may inhibit the approach of DNA
control proteins such as transcription factors. This was confirmed
through a docking study involving sibiromycin and the GAL4 transcription
factor, and these results could explain the significantly enhanced
cytotoxicity of sibiromycin compared to other PBD family members without
bulky C7-substituents
An Extended Pyrrolobenzodiazepine–Polyamide Conjugate with Selectivity for a DNA Sequence Containing the ICB2 Transcription Factor Binding Site
The
binding of nuclear factor Y (NF-Y) to inverted CCAAT boxes
(ICBs) within the promoter region of DNA topoisomerase IIα results
in control of cell differentiation and cell cycle progression. Thus,
NF-Y inhibitory small molecules could be employed to inhibit the replication
of cancer cells. A library of pyrrolobenzodiazepine (PBD) C8-conjugates
consisting of one PBD unit attached to tri-heterocyclic polyamide
fragments was designed and synthesized. The DNA-binding affinity and
sequence selectivity of each compound were evaluated in DNA thermal
denaturation and DNase I footprinting assays, and the ability to inhibit
binding of NF-Y to ICB1 and ICB2 was studied using an electrophoretic
mobility shift assay (EMSA). <b>3a</b> was found to be a potent
inhibitor of NF-Y binding, exhibiting a 10-fold selectivity for an
ICB2 site compared to an ICB1-containing sequence, and showing low
nanomolar cytotoxicity toward human tumor cell lines. Molecular modeling
and computational studies have provided details of the covalent attachment
process that leads to formation of the PBD–DNA adduct, and
have allowed the preference of <b>3a</b> for ICB2 to be rationalized
GC-Targeted C8-Linked Pyrrolobenzodiazepine–Biaryl Conjugates with Femtomolar in Vitro Cytotoxicity and in Vivo Antitumor Activity in Mouse Models
DNA binding 4-(1-methyl-1<i>H</i>-pyrrol-3-yl)Âbenzenamine
(MPB) building blocks have been developed that span two DNA base pairs
with a strong preference for GC-rich DNA. They have been conjugated
to a pyrroloÂ[2,1-<i>c</i>]Â[1,4]Âbenzodiazepine (PBD) molecule
to produce C8-linked PBD–MPB hybrids that can stabilize GC-rich
DNA by up to 13-fold compared to AT-rich DNA. Some have subpicomolar
IC<sub>50</sub> values in human tumor cell lines and in primary chronic
lymphocytic leukemia cells, while being up to 6 orders less cytotoxic
in the non-tumor cell line WI38, suggesting that key DNA sequences
may be relevant targets in these ultrasensitive cancer cell lines.
One conjugate, <b>7h</b> (KMR-28-39), which has femtomolar activity
in the breast cancer cell line MDA-MB-231, has significant dose-dependent
antitumor activity in MDA-MB-231 (breast) and MIA PaCa-2 (pancreatic)
human tumor xenograft mouse models with insignificant toxicity at
therapeutic doses. Preliminary studies suggest that <b>7h</b> may sterically inhibit interaction of the transcription factor NF-κB
with its cognate DNA binding sequence