Fluorescent anticancer quinazolines as molecular probes for β-tubulin colchicine site competition assay and visualization of microtubules as intracellular targeting sites

Suzuki, Yumiko; Sawada, Jun‐ichi; Hibner, Paulina; Ishii, Hirosuke; Matsuno, Kenji; Sato, Masayuki; Witulski, B.; Asai, Akira

doi:10.1016/j.dyepig.2017.05.050

Cited by 12 publications

(15 citation statements)

References 34 publications

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“…In contrast, colchicine EC 70 values for microtubule disappearance and mitotic arrest were similar, 74 and 42 nM, respectively. Although PVHD121 and colchicine bound to the same site of tubulin (Suzuki et al, 2017), PVHD121 was different from colchicine in that the elimination of microtubules was much weaker than mitotic arrest activity. The microtubule-destabilizing activity of PVHD121 in cells was equivalent to the in vitro inhibitory activity against tubulin polymerization in terms of concentration (Kuroiwa et al, 2015).…”

Section: Resultsmentioning

confidence: 83%

“…PVHD277 could act as a fluorescent probe to trace its distribution in cultured cells (Supplemental Fig. 7; Suzuki et al, 2017). PVHD277 was similar to PVHD121 with respect to biological activities, such as tubulin-binding, mitotic entry retardation, and mitotic arrest activities, including the mitotic phenotype (Supplemental Fig.…”

Section: Resultsmentioning

confidence: 91%

“…1A) and its fluorescent derivative 1-(4-methoxyphenyl)-1-(2morpholinoquinazolin-4-yl)ethan-1-ol (PVHD277) ( Fig. 6A) had submicromolar anti-cell proliferative EC 50 values in various tumor-derived cell lines (Suzuki et al, 1998(Suzuki et al, , 2017Kuroiwa et al, 2015). We also showed that they directly bound to the colchicine site of tubulin and inhibited tubulin polymerization in vitro (Kuroiwa et al, 2015).…”

Section: Introductionmentioning

confidence: 80%

“…Chemicals and Antibodies. PVHD121 and PVHD277 were prepared as previously described (Suzuki et al, 1998(Suzuki et al, , 2017Kuroiwa et al, 2015). The purity of the compounds was confirmed on the basis of NMR spectra and melting points (.95%).…”

Section: Methodsmentioning

confidence: 99%

“…The medium was supplemented with 50 mM HEPES (pH 7.4) and BackDrop Background Suppressor (Molecular Probes). Fluorescence images were obtained using a DP30BW CCD camera coupled to an IX71 microscope (Olympus) as previously described (Sawada et al, 2016;Suzuki et al, 2017). The images were colored and superimposed using Adobe Photoshop.…”

Section: Methodsmentioning

confidence: 99%

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Selective Inhibition of Spindle Microtubules by a Tubulin-Binding Quinazoline Derivative

et al. 2019

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In the research field of tubulin-binding agents for the development of anticancer agents, hidden targets are emerging as a problem in understanding the exact mechanisms of actions. The quinazoline derivative 1-(4-methoxyphenyl)-1-(quinazolin-4yl)ethan-1-ol (PVHD121) has anti-cell proliferative activity and inhibits tubulin polymerization by binding to the colchicine site of tubulin. However, the molecular mechanism of action of PVHD121 in cells remains unclear. Here, we demonstrate that PVHD121 delays mitotic entry and efficiently causes mitotic arrest with spindle checkpoint activation, leading to subsequent cell death. The dominant phenotype induced by PVHD121 was aberrant spindles with robust microtubules and unseparated centrosomes. The microtubules were radially distributed, and their ends appeared to adhere to kinetochores, and not to centrosomes. Extensive inhibition by high concentrations of PVHD121 eliminated all microtubules from cells. PVHD277 [1-(4-methoxyphenyl)-1-(2-morpholinoquinazolin-4-yl)ethan-1-ol], a PVHD121 derivative with fluorescence, tended to localize close to the centrosomes when cells prepared to enter mitosis. Our results show that PVHD121 is an antimitotic agent that selectively disturbs microtubule formation at centrosomes during mitosis. This antimitotic activity can be attributed to the targeting of centrosome maturation in addition to the interference with microtubule dynamics. Due to its unique bioactivity, PVHD121 is a potential tool for studying the molecular biology of mitosis and a potential lead compound for the development of anticancer agents. SIGNIFICANCE STATEMENT Many tubulin-binding agents have been developed as potential anticancer agents. The aim of this study was to understand the subcellular molecular actions of a quinazoline derivative tubulinbinding agent, 1-(4-methoxyphenyl)-1-(quinazolin-4-yl)ethan-1ol (PVHD121). As expected from its binding activity to tubulin, PVHD121 caused aberrant spindles and inhibited mitotic progression. However, in addition to tubulin, PVHD121 also targeted an unexpected biomolecule involved in centrosome maturation. Due to targeting the biomolecule just before entering mitosis, PVHD121 preferentially inhibited centrosome-derived microtubules rather than chromosome-derived microtubules during spindle formation. This study not only revealed the molecular action of PVHD121 in cells but also emphasized the importance of considering possible tubulin-independent effects of tubulinbinding agents via hidden targeted biomolecules for future use.

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Section: Resultsmentioning

confidence: 83%

Section: Resultsmentioning

confidence: 91%

Section: Introductionmentioning

confidence: 80%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Selective Inhibition of Spindle Microtubules by a Tubulin-Binding Quinazoline Derivative

et al. 2019

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Synthesis and Structure‐Photophysics Evaluation of 2‐N‐Amino‐quinazolines: Small Molecule Fluorophores for Solution and Solid State

Motoyama

Doan

Hibner-Kulicka

et al. 2021

Chemistry An Asian Journal

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2‐N‐aminoquinazolines were prepared by consecutive SNAr functionalization. X‐ray structures display the nitrogen lone pair of the 2‐N‐morpholino group in conjugation with the electron deficient quinazoline core and thus representing electronic push‐pull systems. 2‐N‐aminoquinazolines show a positive solvatochromism and are fluorescent in solution and in solid state with quantum yields up to 0.73. Increase in electron donor strength of the 2‐amino substituent causes a red‐shift of the intramolecular charge transfer (ICT) band (300–400 nm); whereas the photoluminescence emission maxima (350–450 nm) is also red‐shifted significantly along with an enhancement in photoluminescence efficiency. HOMO‐LUMO energies were estimated by a combination of electrochemical and photophysical methods and correlate well to those obtained by computational methods. ICT properties are theoretically attributed to an excitation to Rydberg‐MO in SAC‐CI method, which can be interpreted as n‐π* excitation. 7‐Amino‐2‐N‐morpholino‐4‐methoxyquinazoline responds to acidic conditions with significant increases in photoluminescence intensity revealing a new turn‐on/off fluorescence probe.

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One‐Pot Synthesis and Photophysical Studies of Styryl‐Based Benzo[f]pyrazolo[3,4‐b]quinoline and Indeno[2,1‐b]pyrazolo[4,3‐e]pyridines

et al. 2018

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A new family of extended π‐conjugated dihydrobenzopyrazoloquinoline (DBPQ) and dihydroindenopyrazolopyridine (DIPP) molecules were synthesized, and their photophysical and electrochemical properties were studied. The DBPQ derivatives showed significantly higher molar absorption coefficients, and their absorption and emission wavelengths were red‐shifted with respect to those of the corresponding DIPP derivatives in chloroform solution. Dyes with no substitution and methoxy, nitro, and fluoro‐substituted derivatives showed poor dependence on solvent polarity. At the same time, DBPQ and DIPP derivatives with dimethylamino groups exhibited strong solvatochromism in the emission spectra. Due to the presence of a vinyl link with a cylindrical π‐cloud on the donor (dimethylamino group) molecule, they showed efficient positive solvatochromism with high quantum yields on increasing solvent polarity from non‐polar to polar solvents. Aggregation‐induced emission (AIE) properties were also investigated. AIE studies show that the emission of DBPQ and DIPP derivatives in the presence of THF/H2O mixture increased due to aggregation. However, DBPQ and DIPP derivatives were still strongly emissive in the solid state. Dimethylamino substituted derivatives exhibited acidochromism, which was caused by the addition of TFA and pTsA leading to prominent red‐shifts in the emission spectra owing to the protonation at the pyridine segment.

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Fluorescent anticancer quinazolines as molecular probes for β-tubulin colchicine site competition assay and visualization of microtubules as intracellular targeting sites

Cited by 12 publications

References 34 publications

Selective Inhibition of Spindle Microtubules by a Tubulin-Binding Quinazoline Derivative

Selective Inhibition of Spindle Microtubules by a Tubulin-Binding Quinazoline Derivative

Synthesis and Structure‐Photophysics Evaluation of 2‐N‐Amino‐quinazolines: Small Molecule Fluorophores for Solution and Solid State

One‐Pot Synthesis and Photophysical Studies of Styryl‐Based Benzo[f]pyrazolo[3,4‐b]quinoline and Indeno[2,1‐b]pyrazolo[4,3‐e]pyridines

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