Identification of pyrrolopyrimidine derivative PP-13 as a novel microtubule-destabilizing agent with promising anticancer properties

Abstract

Despite the emergence of targeted therapies and immunotherapy, chemotherapy remains the gold-standard for the treatment of most patients with solid malignancies. Spindle poisons that interfere with microtubule dynamics are commonly used in chemotherapy drug combinations. However, their troublesome side effects and the emergence of chemoresistance highlight the need for identifying alternative agents. We performed a high throughput cell-based screening and selected a pyrrolopyrimidine molecule (named PP-13). In the present study, we evaluated its anticancer properties in vitro and in vivo. We showed that PP-13 exerted cytotoxic effects on various cancer cells, including those resistant to current targeted therapies and chemotherapies. PP-13 induced a transient mitotic blockade by interfering with both mitotic spindle organization and microtubule dynamics and finally led to mitotic slippage, aneuploidy and direct apoptotic death. PP-13 was identified as a microtubule-targeting agent that binds directly to the colchicine site in β-tubulin. Interestingly, PP-13 overcame the multidrug-resistant cancer cell phenotype and significantly reduced tumour growth and metastatic invasiveness without any noticeable toxicity for the chicken embryo in vivo. Overall, PP-13 appears to be a novel synthetic microtubule inhibitor with interesting anticancer properties and could be further investigated as a potent alternative for the management of malignancies including chemoresistant ones.

Synthesis and Anticancer Activity of Mitotic-Specific 3,4-Dihydropyridine-2(1H)-thiones

“…Polyploidy is also typical for cells blocked in mitosis by colchicine (described as ‘‘c-mitosis’’ or ‘‘colchicine-mitosis’’), as a results of partial or complete absence of spindle apparatus following the breakdown of nuclear envelope, condensed chromosomes, and undivided centromeres [ 30 ]. Our data are consistent with Gilson et al study, where after initial accumulation of H358 and HeLa cells in mitosis as a result of pyrrolopyrimidine (PP-13) treatment, the percentage of G2/M arrested cells was gradually decreasing, concomitantly with the accumulation in the sub-G1 and polyploid fraction [ 31 ]. As mentioned above, inhibition of tubulin polymerization and mitotic spindle formation may be one of the reasons for polyploid nuclei formation.…”

“…We could describe the three most common abnormal phenotypes: (1) bipolar spindles with chromosomes arrangement in the central metaphase and additional “cloud” of chromosomes around of centrioles (misalignment chromosomes), (2) monoastral spindles, (3) multipolar asymmetric asters ( Figure 5 ). Observed aberrations were previously associated with strong defects in chromosome congression and alignment, and may be connected with prometaphase blockade [ 31 ]. Multipolar and “bipolar misaligned” spindles were described by Jaunky et al in cells treated with colchicine and C75 (new CBSIs), depending on concentration and (in the case of C75) cell line used.…”

Unprecedented inhibition of P-gp activity by a novel ruthenium-cyclopentadienyl compound bearing a bipyridine-biotin ligand

“…Cell lines and culture conditions-To evaluate the selectivity of the compounds on other ABC transporters, NIH3T3 parental cell line and NIH3T3/ABCB1 drug resistant cell line transfected with human MDR1/A-G185, purchased from American Type Culture Collection (Manassas, VA) were used, as previously described. [31] HEK293 (Human embryonic kidney cell) were used to express the pcDNA3.1-hABCG2 plasmid [32] and Flp-In™−293 cells to express ABCC1 and ABCC2 genes transfected by electroporation using Neon® Transfection System (ThermoFisher scientific) with pcDNA5-FRT-ABCC1 or pcDNA5-FRT-ABCC2 respectively as previously described [33]. MCF7 and MDA-MB-231 breast cancer human tumor cell lines were obtained from ATCC.…”

Novel Bis- and Mono-Pyrrolo[2,3-d]pyrimidine and Purine Derivatives: Synthesis, Computational Analysis and Antiproliferative Evaluation

“…Pyrrolo [2,3-d]pyrimidine sulfonamides were recently found to act as cytotoxic agents in hypoxia via inhibition of transmembrane carbonic anhydrases [37]. A 4-(benzylamino)-pyrrolo [2,3-d]pyrimidine derivative exerted potent antitumour effects in vivo and induced mitotic cell blockade by impairing both mitotic microtubule organization and dynamics [38]. To overcome multidrug resistance in cancer patients, pyrrolo [2,3-d]pyrimidines and purine derivatives with high lipophilicity and molecular weight were developed as potent and selective inhibitors of multidrug resistance-associated protein 1 (MRP1, ABCC1) associated with non-response to chemotherapy in different cancers [39].…”