6 research outputs found
Plasmin-Binding Tripeptide-Decorated Liposomes Loading Pyrazolo[3,4‑<i>d</i>]pyrimidines for Targeting Hepatocellular Carcinoma
Hepatocellular
carcinoma (HCC) is one of the most fatal cancer
types worldwide. HCC cells were proved to overexpress c-Src and Sgk1,
a tyrosine and a serine-threonine kinase, respectively, whose role
is crucial for the development and progression of the tumor. Pyrazolo[3,4-<i>d</i>]pyrimidine derivatives are a class of tyrosine kinase
inhibitors that have shown good activity against HepG2. HCC cells
were also proved to overexpress plasmin, which is localized on the
cell surface bound to its receptors. In this study, a tripeptide with
sequence d-Ala-Phe-Lys, which binds a specific reactive site
of plasmin, was synthesized and characterized. This tripeptide was
used to decorate liposomes encapsulating three selected pyrazolo[3,4-<i>d</i>]pyrimidines. Liposomes bearing tripeptide have been characterized,
not showing remarkable differences with respect to the corresponding
tripeptide-free liposomes. <i>In vitro</i> HepG2 cell uptake
profiles and cytotoxicities showed that the presence of the tripeptide
on the liposomal membrane surface improves the cell-penetrating ability
of liposomes and increases the activity of two of the three tested
compounds
Identification of New Fyn Kinase Inhibitors Using a FLAP-Based Approach
The
abnormal activity of Fyn tyrosine kinase has been shown to
be related to various human cancers. Furthermore, its involvement
in signaling pathways that lead to severe pathologies, such as Alzheimer’s
and Parkinson’s diseases, has also been demonstrated, thus
making Fyn an attractive target for the discovery of potential novel
therapeutics for brain pathologies and tumors. In this study we evaluated
the reliability of various screening approaches based on the FLAP
software. By the application of the best procedure, the virtual screening
workflow was used to filter the Gold and Platinum database from Asinex
to identify new Fyn inhibitors. Enzymatic assays revealed that among
the eight top-scoring compounds five proved to efficiently inhibit
Fyn activity with IC<sub>50</sub> values in the micromolar range.
These results demonstrate the validity of the methodologies we followed.
Furthermore, the five active compounds herein described may be considered
as interesting leads for the development of new and more efficient
Fyn inhibitors
In Silico Identification and Biological Evaluation of Novel Selective Serum/Glucocorticoid-Inducible Kinase 1 Inhibitors Based on the Pyrazolo-Pyrimidine Scaffold
The
serum/glucocorticoid-inducible kinase 1 (Sgk1) has demonstrated
antiapoptotic function and the capability to regulate cell survival,
proliferation, and differentiation. A pivotal role of Sgk1 in carcinogenesis
and in resistance to anticancer therapy has been suggested. With the
aim of identifying new Sgk1 modulators, 322 pyrazolo-pyrimidine derivatives
have been virtually screened with respect to a crystallographic model
of Sgk1. The top five ranked compounds have been evaluated demonstrating
Sgk1 inhibition in vitro and selectivity compared to RAC-alpha serine/threonine-protein
kinase (Akt1)
Design, Synthesis, and Biological Evaluation of Pyrazolo[3,4‑<i>d</i>]pyrimidines Active in Vivo on the Bcr-Abl T315I Mutant
Starting from our in-house library
of pyrazolo[3,4-<i>d</i>]pyrimidines, a cross-docking simulation
was conducted on Bcr-Abl
T315I mutant. Among the selected compounds (<b>2a</b>–<b>e</b>), the 4-bromo derivative <b>2b</b> showed the best
activity against the Bcr-Abl T315I mutant. Deeper computational studies
highlighted the importance of the bromine atom in the para position
of the N1 side chain phenyl ring for the interaction with the T315I
mutant. A series of 4-bromo derivatives was thus synthesized and biologically
evaluated. Compound <b>2j</b> showed a good balance of different
ADME properties, high activity in cell-free assays, and a submicromolar
potency against T315I Bcr-Abl expressing cells. In addition, it was
converted into a water-soluble formulation by liposome encapsulation,
preserving a good activity on leukemic T315I cells and avoiding the
use of DMSO as solubilizing agent. In vivo studies on mice inoculated
with 32D-T315I cells and treated with <b>2j</b> showed a more
than 50% reduction
in tumor volumes
Combining X‑ray Crystallography and Molecular Modeling toward the Optimization of Pyrazolo[3,4‑<i>d</i>]pyrimidines as Potent c‑Src Inhibitors Active in Vivo against Neuroblastoma
c-Src
is a tyrosine kinase belonging to the Src-family kinases.
It is overexpressed and/or hyperactivated in a variety of cancer cells,
thus its inhibition has been predicted to have therapeutic effects
in solid tumors. Recently, the pyrazolo[3,4-<i>d</i>]pyrimidine <b>3</b> was reported as a dual c-Src/Abl inhibitor. Herein we describe
a multidisciplinary drug discovery approach for the optimization of
the lead <b>3</b> against c-Src. Starting from the X-ray crystal
structure of c-Src in complex with <b>3</b>, Monte Carlo free
energy perturbation calculations were applied to guide the design
of c-Src inhibitors with improved activities. As a result, the introduction
of a <i>meta</i> hydroxyl group on the C4 anilino ring was
computed to be particularly favorable. The potency of the synthesized
inhibitors was increased with respect to the starting lead <b>3</b>. The best identified compounds were also found active in the inhibition
of neuroblastoma cell proliferation. Furthermore, compound <b>29</b> also showed in vivo activity in xenograft model using SH-SY5Y cells
Prodrugs of Pyrazolo[3,4‑<i>d</i>]pyrimidines: From Library Synthesis to Evaluation as Potential Anticancer Agents in an Orthotopic Glioblastoma Model
Pyrazolo[3,4-<i>d</i>]pyrimidines
are potent protein
kinase inhibitors with promising antitumor activity but suboptimal
aqueous solubility, consequently worth being further optimized. Herein,
we present the one-pot two-step procedure for the synthesis of a set
of pyrazolo[3,4-<i>d</i>]pyrimidine prodrugs (<b>1a</b>−<b>8a</b> and <b>9a</b>−<b>e</b>) with higher aqueous solubility and enhanced pharmacokinetic and
therapeutic properties. ADME studies demonstrated for the most promising
prodrugs a better aqueous solubility, a favorable hydrolysis in human
and murine serum, and an increased ability to cross cell membranes
with respect to the parental drugs, explaining their better 24 h <i>in vitro</i> cytotoxicity against human glioblastoma U87 cell
line. Finally, the <b>4–4a</b> couple of drug/prodrug
was also evaluated <i>in vivo</i>, revealing a profitable
pharmacokinetic profile of the prodrug associated with a good efficacy.
The application of the prodrug approach demonstrated to be a successful
strategy for improving aqueous solubility of the parental drugs, determining
a positive impact also in their biological efficacy