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A novel orally active water-soluble inhibitor of human glutathione transferase exerts a potent and selective antitumor activity against human melanoma xenografts.

De Luca A, Rotili D, Carpanese D, Lenoci A, Calderan L, Scimeca M, Mai A, Bonanno E, Rosato A, Geroni C, Quintieri L, Caccuri AM - Oncotarget (2015)

Bottom Line: Interestingly, the cellular response to MC3181 was cell-type-specific; the compound triggered a JNK-dependent apoptosis in the BRAF-V600E-mutated A375 cells, while it induced morphological changes together with an increase in melanogenesis in BRAF wild-type SK23-MEL cells.MC3181 exhibited a remarkable therapeutic activity against BRAF-V600E-mutant xenografts, both after intravenous and oral administration.Outstandingly, no treatment-related signs of toxicity were observed both in healthy and tumor-bearing mice after single and repeated administrations.

View Article: PubMed Central - PubMed

Affiliation: The NAST Centre for Nanoscience & Nanotechnology & Innovative Instrumentation, University of Tor Vergata, 00133 Rome, Italy.

ABSTRACT
We designed and synthesized two novel nitrobenzoxadiazole (NBD) analogues of the anticancer agent 6-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thio)hexan-1-ol (NBDHEX). The new compounds, namely MC3165 and MC3181, bear one and two oxygen atoms within the hydroxy-containing alkyl chain at the C4 position of the NBD scaffold, respectively. This insertion did not alter the chemical reactivity with reduced glutathione, while it conferred a remarkable increase in water solubility. MC3181 was more selective than NBDHEX towards the target protein, glutathione transferase P1-1, and highly effective in vitro against a panel of human melanoma cell lines, with IC50 in the submicromolar-low micromolar range. Interestingly, the cellular response to MC3181 was cell-type-specific; the compound triggered a JNK-dependent apoptosis in the BRAF-V600E-mutated A375 cells, while it induced morphological changes together with an increase in melanogenesis in BRAF wild-type SK23-MEL cells. MC3181 exhibited a remarkable therapeutic activity against BRAF-V600E-mutant xenografts, both after intravenous and oral administration. Outstandingly, no treatment-related signs of toxicity were observed both in healthy and tumor-bearing mice after single and repeated administrations. Taken together, these results indicate that MC3181 may represent a potential novel therapeutic opportunity for BRAF-mutated human melanoma, while being safe and water-soluble and thus overcoming all the critical aspects of NBDHEX in vivo.

No MeSH data available.


Related in: MedlinePlus

MC3181 triggers JNK-dependent apoptosis in A375 cells andmorphological changes in SK23-MEL cells(A) The percentage of early apototic, late apoptotic, andnecrotic A375 cells was evaluated by cytofluorimetric analysis ofAnnexin V versus PI staining, after 24 and 48 hrs incubation with 10μM MC3181. (B, C) Pre-incubation of A375 cell linewith the JNK inhibitor SP600125 (20 μM) significantly reduced thepercentage of Sub-G1 (apoptotic) cells, and strongly suppressedcaspase-3 activation. (D) Percentage of early apototic,late apoptotic, and necrotic cells and (E) of sub-G1 phasepopulation in SK23-MEL cells following 48 hrs exposure to 7 μMMC3181. (F) Phase-contrast microscopy images (10Xmagnification, 3X digital magnification) show SK23-MEL cells with atypical triangular dendritic/spindle shape in control cultures, whilstbipolar spindle morphology is induced by 48 hrs incubation with MC3181(7 μM). An increase of melanin content and tyrosinase activitywere also observed in SK23-MEL cells after 48 hrs incubation withMC3181. Data represent means ± SD of three independentexperiments.
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Figure 5: MC3181 triggers JNK-dependent apoptosis in A375 cells andmorphological changes in SK23-MEL cells(A) The percentage of early apototic, late apoptotic, andnecrotic A375 cells was evaluated by cytofluorimetric analysis ofAnnexin V versus PI staining, after 24 and 48 hrs incubation with 10μM MC3181. (B, C) Pre-incubation of A375 cell linewith the JNK inhibitor SP600125 (20 μM) significantly reduced thepercentage of Sub-G1 (apoptotic) cells, and strongly suppressedcaspase-3 activation. (D) Percentage of early apototic,late apoptotic, and necrotic cells and (E) of sub-G1 phasepopulation in SK23-MEL cells following 48 hrs exposure to 7 μMMC3181. (F) Phase-contrast microscopy images (10Xmagnification, 3X digital magnification) show SK23-MEL cells with atypical triangular dendritic/spindle shape in control cultures, whilstbipolar spindle morphology is induced by 48 hrs incubation with MC3181(7 μM). An increase of melanin content and tyrosinase activitywere also observed in SK23-MEL cells after 48 hrs incubation withMC3181. Data represent means ± SD of three independentexperiments.

Mentions: Flow cytometry analysis of cell cycle perturbations induced by MC3181 in A375cells revealed a time-dependent increase in the number of cells blocked in theG2/M phase and a concomitant increase in the amount of cells in the sub-G1 phase(about 27 and 36% increase after 24 and 48 hrs, respectively; Fig. 4B, left panel and Fig. 5, panels B and C). A noticeable cell cycle arrest in theG2/M phase was also observed in MC3181-treated SK23-MEL cells, whereas thedrug-induced increase in the number of cells in sub-G1 phase was less pronouncedthan that recorded in A375 cells (about 10 and 20% increase after 24 and 48 hrsof treatment, respectively; Fig. 4B, rightpanel and Fig. 5, panel E). Thesedifferences translated into a different degree of caspase activation; a strongcaspase-3 activity (Fig. 4C, left panel)was observed in drug-treated A375 cells, while MC3181 induced only a negligibleincrease of proteolitic activity in SK23-MEL cells (Fig. 4C, right panel).


A novel orally active water-soluble inhibitor of human glutathione transferase exerts a potent and selective antitumor activity against human melanoma xenografts.

De Luca A, Rotili D, Carpanese D, Lenoci A, Calderan L, Scimeca M, Mai A, Bonanno E, Rosato A, Geroni C, Quintieri L, Caccuri AM - Oncotarget (2015)

MC3181 triggers JNK-dependent apoptosis in A375 cells andmorphological changes in SK23-MEL cells(A) The percentage of early apototic, late apoptotic, andnecrotic A375 cells was evaluated by cytofluorimetric analysis ofAnnexin V versus PI staining, after 24 and 48 hrs incubation with 10μM MC3181. (B, C) Pre-incubation of A375 cell linewith the JNK inhibitor SP600125 (20 μM) significantly reduced thepercentage of Sub-G1 (apoptotic) cells, and strongly suppressedcaspase-3 activation. (D) Percentage of early apototic,late apoptotic, and necrotic cells and (E) of sub-G1 phasepopulation in SK23-MEL cells following 48 hrs exposure to 7 μMMC3181. (F) Phase-contrast microscopy images (10Xmagnification, 3X digital magnification) show SK23-MEL cells with atypical triangular dendritic/spindle shape in control cultures, whilstbipolar spindle morphology is induced by 48 hrs incubation with MC3181(7 μM). An increase of melanin content and tyrosinase activitywere also observed in SK23-MEL cells after 48 hrs incubation withMC3181. Data represent means ± SD of three independentexperiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4414177&req=5

Figure 5: MC3181 triggers JNK-dependent apoptosis in A375 cells andmorphological changes in SK23-MEL cells(A) The percentage of early apototic, late apoptotic, andnecrotic A375 cells was evaluated by cytofluorimetric analysis ofAnnexin V versus PI staining, after 24 and 48 hrs incubation with 10μM MC3181. (B, C) Pre-incubation of A375 cell linewith the JNK inhibitor SP600125 (20 μM) significantly reduced thepercentage of Sub-G1 (apoptotic) cells, and strongly suppressedcaspase-3 activation. (D) Percentage of early apototic,late apoptotic, and necrotic cells and (E) of sub-G1 phasepopulation in SK23-MEL cells following 48 hrs exposure to 7 μMMC3181. (F) Phase-contrast microscopy images (10Xmagnification, 3X digital magnification) show SK23-MEL cells with atypical triangular dendritic/spindle shape in control cultures, whilstbipolar spindle morphology is induced by 48 hrs incubation with MC3181(7 μM). An increase of melanin content and tyrosinase activitywere also observed in SK23-MEL cells after 48 hrs incubation withMC3181. Data represent means ± SD of three independentexperiments.
Mentions: Flow cytometry analysis of cell cycle perturbations induced by MC3181 in A375cells revealed a time-dependent increase in the number of cells blocked in theG2/M phase and a concomitant increase in the amount of cells in the sub-G1 phase(about 27 and 36% increase after 24 and 48 hrs, respectively; Fig. 4B, left panel and Fig. 5, panels B and C). A noticeable cell cycle arrest in theG2/M phase was also observed in MC3181-treated SK23-MEL cells, whereas thedrug-induced increase in the number of cells in sub-G1 phase was less pronouncedthan that recorded in A375 cells (about 10 and 20% increase after 24 and 48 hrsof treatment, respectively; Fig. 4B, rightpanel and Fig. 5, panel E). Thesedifferences translated into a different degree of caspase activation; a strongcaspase-3 activity (Fig. 4C, left panel)was observed in drug-treated A375 cells, while MC3181 induced only a negligibleincrease of proteolitic activity in SK23-MEL cells (Fig. 4C, right panel).

Bottom Line: Interestingly, the cellular response to MC3181 was cell-type-specific; the compound triggered a JNK-dependent apoptosis in the BRAF-V600E-mutated A375 cells, while it induced morphological changes together with an increase in melanogenesis in BRAF wild-type SK23-MEL cells.MC3181 exhibited a remarkable therapeutic activity against BRAF-V600E-mutant xenografts, both after intravenous and oral administration.Outstandingly, no treatment-related signs of toxicity were observed both in healthy and tumor-bearing mice after single and repeated administrations.

View Article: PubMed Central - PubMed

Affiliation: The NAST Centre for Nanoscience & Nanotechnology & Innovative Instrumentation, University of Tor Vergata, 00133 Rome, Italy.

ABSTRACT
We designed and synthesized two novel nitrobenzoxadiazole (NBD) analogues of the anticancer agent 6-((7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)thio)hexan-1-ol (NBDHEX). The new compounds, namely MC3165 and MC3181, bear one and two oxygen atoms within the hydroxy-containing alkyl chain at the C4 position of the NBD scaffold, respectively. This insertion did not alter the chemical reactivity with reduced glutathione, while it conferred a remarkable increase in water solubility. MC3181 was more selective than NBDHEX towards the target protein, glutathione transferase P1-1, and highly effective in vitro against a panel of human melanoma cell lines, with IC50 in the submicromolar-low micromolar range. Interestingly, the cellular response to MC3181 was cell-type-specific; the compound triggered a JNK-dependent apoptosis in the BRAF-V600E-mutated A375 cells, while it induced morphological changes together with an increase in melanogenesis in BRAF wild-type SK23-MEL cells. MC3181 exhibited a remarkable therapeutic activity against BRAF-V600E-mutant xenografts, both after intravenous and oral administration. Outstandingly, no treatment-related signs of toxicity were observed both in healthy and tumor-bearing mice after single and repeated administrations. Taken together, these results indicate that MC3181 may represent a potential novel therapeutic opportunity for BRAF-mutated human melanoma, while being safe and water-soluble and thus overcoming all the critical aspects of NBDHEX in vivo.

No MeSH data available.


Related in: MedlinePlus