Browsing by Author "Chtcherbinine, Mikhail"

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    Development of 2,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one inhibitors of Aldehyde Dehydrogenase 1A (ALDH1A) as potential adjuncts to ovarian cancer chemotherapy
    (Elsevier, 2021-02) Huddle, Brandt C.; Grimley, Edward; Chtcherbinine, Mikhail; Buchman, Cameron D.; Takahashi, Cyrus; Debnath, Bikash; McGonigal, Stacy C.; Mao, Shuai; Li, Siwei; Felton, Jeremy; Pan, Shu; Wen, Bo; Sun, Duxin; Neamati, Nouri; Buckanovich, Ronald J.; Hurley, Thomas D.; Larsen, Scott D.; Biochemistry and Molecular Biology, School of Medicine
    There is strong evidence that inhibition of one or more Aldehyde Dehydrogenase 1A (ALDH1A) isoforms may be beneficial in chemotherapy-resistant ovarian cancer and other tumor types. While many previous efforts have focused on development of ALDH1A1 selective inhibitors, the most deadly ovarian cancer subtype, high-grade serous (HGSOC), exhibits elevated expression of ALDH1A3. Herein, we report continued development of pan-ALDH1A inhibitors to assess whether broad spectrum ALDH1A inhibition is an effective adjunct to chemotherapy in this critical tumor subtype. Optimization of the CM39 scaffold, aided by metabolite ID and several new ALDH1A1 crystal structures, led to improved biochemical potencies, improved cellular ALDH inhibition in HGSOC cell lines, and substantial improvements in microsomal stability culminating in orally bioavailable compounds. We demonstrate that two compounds 68 and 69 are able to synergize with chemotherapy in a resistant cell line and patient-derived HGSOC tumor spheroids, indicating their suitability for future in vivo proof of concept experiments.
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    Development of substituted benzimidazoles as inhibitors of Human Aldehyde Dehydrogenase 1A Isoenzymes
    (Elsevier, 2024) Takahashi, Cyrus; Chtcherbinine, Mikhail; Huddle, Brandt C.; Wilson, Michael W.; Emmel, Timothy; Hohlman, Robert M.; McGonigal, Stacy; Buckanovich, Ronald J.; Larsen, Scott D.; Hurley, Thomas D.; Biochemistry and Molecular Biology, School of Medicine
    Aldehyde dehydrogenase 1A (ALDH1A) isoforms may be a useful target for overcoming chemotherapy resistance in high-grade serous ovarian cancer (HGSOC) and other solid tumor cancers. However, as different cancers express different ALDH1A isoforms, isoform selective inhibitors may have a limited therapeutic scope. Furthermore, resistance to an ALDH1A isoform selective inhibitor could arise via induction of expression of other ALDH1A isoforms. As such, we have focused on the development of pan-ALDH1A inhibitors, rather than on ALDH1A isoform selective compounds. Herein, we report the development of a new group of pan-ALDH1A inhibitors to assess whether broad spectrum ALDH1A inhibition is an effective adjunct to chemotherapy in HGSOC. Optimization of the CM10 scaffold, aided by ALDH1A1 crystal structures, led to improved biochemical potencies, improved cellular efficacy as demonstrated by reduction in ALDEFLUOR signal in HGSOC cells, and substantial improvements in liver microsomal stability. Based on this work we identified two compounds 17 and 25 suitable for future in vivo proof of concept experiments.
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    Kinetic and Structural Characterization of Isoenzyme-Selective Aldehyde Dehydrogenase 1A Inhibitors
    (2016) Chtcherbinine, Mikhail; Hurley, Thomas D.; Georgiadis, Millie M.; Wells, Clark D.
    The human aldehyde dehydrogenase superfamily consists of 19 distinct genetic loci that play key roles in both health and disease. Aldehyde dehydrogenases are primarily involved in the metabolism of reactive aldehyde substrates; the ALDH1A subfamily, in particular, metabolizes retinaldehyde and is involved in a pathway regulating tissue differentiation, cell proliferation, and apoptosis. Recently, ALDH1 isoenzymes have been implicated as significant elements in cancer progression. ALDH1 activity has been used as a marker of cancer stem cells, a subpopulation of cancer stem cells with high drug resistance, proliferative potential, and ability to differentiate into multiple cell types. In accordance with this, ALDH1 activity and expression has been shown to correlate with lower survival, increased chemoresistance, and increased chance of relapse in multiple solid cancer types, including breast, ovarian, lung, and colorectal. Despite the clear relevance of ALDH1 enzymes in cancer, the specific roles of individual isoenzymes are unclear. Isoenzyme-selective small molecule modulators of the ALDH1A subfamily would allow the probing of the function of individual isoenzymes in healthy and disease states. Two ALDH1A1 inhibitors, CM38 and C10, were previously identified in a high-throughput screen. In this study, CM38, an ALDH1A1-selective inhibitor, and CM10, an ALDH1A inhibitor, were characterized using kinetic assays, structural biology, and cell culture experiments. A structure-activity relationship was built for each series, and an X-ray crystallography structure was used to determine the binding mode. These approaches allowed the investigation of the ALDH1A active site and identification of structural features that can be used to design and improve selective modulators of this subfamily. CM38 and CM10 were also tested in a breast cancer cell line to determine their efficacy in a cellular environment. While the CM38 series showed warning signs of potential off-target toxicity, members of the CM10 compound series showed excellent initial characteristics as potential chemical tools. The results of this study may be useful in the design of new chemical tools to delineate the functions of individual ALDH1 isoenzymes in cancer biology, as well as in the development of drugs to selectively target cancer stem cells.
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    A Novel ALDH1A1 Inhibitor Blocks Platinum-Induced Senescence and Stemness in Ovarian Cancer
    (MDPI, 2022-07-15) Muralikrishnan, Vaishnavi; Fang, Fang; Given, Tyler C.; Podicheti, Ram; Chtcherbinine, Mikhail; Metcalfe, Tara X.; Sriramkumar, Shruthi; O’Hagan, Heather M.; Hurley, Thomas D.; Nephew, Kenneth P.; Medical and Molecular Genetics, School of Medicine
    Ovarian cancer is a deadly disease attributed to late-stage detection as well as recurrence and the development of chemoresistance. Ovarian cancer stem cells (OCSCs) are hypothesized to be largely responsible for the emergence of chemoresistant tumors. Although chemotherapy may initially succeed at decreasing the size and number of tumors, it leaves behind residual malignant OCSCs. In this study, we demonstrate that aldehyde dehydrogenase 1A1 (ALDH1A1) is essential for the survival of OCSCs. We identified a first-in-class ALDH1A1 inhibitor, compound 974, and used 974 as a tool to decipher the mechanism of stemness regulation by ALDH1A1. The treatment of OCSCs with 974 significantly inhibited ALDH activity, the expression of stemness genes, and spheroid and colony formation. An in vivo limiting dilution assay demonstrated that 974 significantly inhibited CSC frequency. A transcriptomic sequencing of cells treated with 974 revealed a significant downregulation of genes related to stemness and chemoresistance as well as senescence and the senescence-associated secretory phenotype (SASP). We confirmed that 974 inhibited the senescence and stemness induced by platinum-based chemotherapy in functional assays. Overall, these data establish that ALDH1A1 is essential for OCSC survival and that ALDH1A1 inhibition suppresses chemotherapy-induced senescence and stemness. Targeting ALDH1A1 using small-molecule inhibitors in combination with chemotherapy therefore presents a promising strategy to prevent ovarian cancer recurrence and has the potential for clinical translation.
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    A Pan-ALDH1A Inhibitor Induces Necroptosis in Ovarian Cancer Stem-like Cells
    (Elsevier, 2019-03-12) Chefetz, Ilana; Grimley, Edward; Yang, Kun; Hong, Linda; Vinogradova, Ekaterina V.; Suciu, Radu; Kovalenko, Ilya; Karnak, David; Morgan, Cynthia A.; Chtcherbinine, Mikhail; Buchman, Cameron; Huddle, Brandt; Barraza, Scott; Morgan, Meredith; Bernstein, Kara A.; Yoon, Euisik; Lombard, David B.; Bild, Andrea; Mehta, Geeta; Romero, Iris; Chiang, Chun-Yi; Landen, Charles; Cravatt, Benjamin; Hurley, Thomas D.; Larsen, Scott D.; Buckanovich, Ronald J.; Department of Biochemistry and Molecular Biology, School of Medicine
    Summary Ovarian cancer is typified by the development of chemotherapy resistance. Chemotherapy resistance is associated with high aldehyde dehydrogenase (ALDH) enzymatic activity, increased cancer “stemness,” and expression of the stem cell marker CD133. As such, ALDH activity has been proposed as a therapeutic target. Although it remains controversial which of the 19 ALDH family members drive chemotherapy resistance, ALDH1A family members have been primarily linked with chemotherapy resistant and stemness. We identified two ALDH1A family selective inhibitors (ALDH1Ai). ALDH1Ai preferentially kills CD133+ ovarian cancer stem-like cells (CSCs). ALDH1Ai induce necroptotic CSC death, mediated, in part, by the induction of mitochondrial uncoupling proteins and reduction in oxidative phosphorylation. ALDH1Ai is highly synergistic with chemotherapy, reducing tumor initiation capacity and increasing tumor eradication in vivo. These studies link ALDH1A with necroptosis and confirm the family as a critical therapeutic target to overcome chemotherapy resistance and improve patient outcomes.
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    Structure-Based Optimization of a Novel Class of Aldehyde Dehydrogenase 1A (ALDH1A) Subfamily-Selective Inhibitors as Potential Adjuncts to Ovarian Cancer Chemotherapy
    (American Chemical Society, 2018-10-11) Huddle, Brandt C.; Grimley, Edward; Buchman, Cameron D.; Chtcherbinine, Mikhail; Debnath, Bikash; Mehta, Pooja; Yang, Kun; Morgan, Cynthia A.; Li, Siwei; Felton, Jeremy; Sun, Duxin; Mehta, Geeta; Neamati, Nouri; Buckanovich, Ronald J.; Hurley, Thomas D.; Larsen, Scott D.; Biochemistry and Molecular Biology, School of Medicine
    Aldehyde dehydrogenase (ALDH) activity is commonly used as a marker to identify cancer stem-like cells. The three ALDH1A isoforms have all been individually implicated in cancer stem-like cells and in chemoresistance; however, which isoform is preferentially expressed varies between cell lines. We sought to explore the structural determinants of ALDH1A isoform selectivity in a series of small-molecule inhibitors in support of research into the role of ALDH1A in cancer stem cells. An SAR campaign guided by a cocrystal structure of the HTS hit CM39 (7) with ALDH1A1 afforded first-in-class inhibitors of the ALDH1A subfamily with excellent selectivity over the homologous ALDH2 isoform. We also discovered the first reported modestly selective single isoform 1A2 and 1A3 inhibitors. Two compounds, 13g and 13h, depleted the CD133+ putative cancer stem cell pool, synergized with cisplatin, and achieved efficacious concentrations in vivo following IP administration. Compound 13h additionally synergized with cisplatin in a patient-derived ovarian cancer spheroid model.
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    Structure-Based Optimization of a Novel Class of Aldehyde Dehydrogenase 1A (ALDH1A) Subfamily-Selective Inhibitors as Potential Adjuncts to Ovarian Cancer Chemotherapy
    (ACS, 2018-09) Huddle, Brandt C.; Grimley, Edward; Buchman, Cameron D.; Chtcherbinine, Mikhail; Debnath, Bikash; Mehta, Pooja; Yang, Kun; Morgan, Cynthia A.; Li, Siwei; Felton, Jeremy; Sun, Duxin; Mehta, Geeta; Neamati, Nouri; Buckanovich, Ronald J.; Hurley, Thomas D.; Larsen, Scott D.; Biochemistry and Molecular Biology, School of Medicine
    Aldehyde dehydrogenase (ALDH) activity is commonly used as a marker to identify cancer stem-like cells. The three ALDH1A isoforms have all been individually implicated in cancer stem-like cells and in chemoresistance; however, which isoform is preferentially expressed varies between cell lines. We sought to explore the structural determinants of ALDH1A isoform selectivity in a series of small-molecule inhibitors in support of research into the role of ALDH1A in cancer stem cells. An SAR campaign guided by a cocrystal structure of the HTS hit CM39 (7) with ALDH1A1 afforded first-in-class inhibitors of the ALDH1A subfamily with excellent selectivity over the homologous ALDH2 isoform. We also discovered the first reported modestly selective single isoform 1A2 and 1A3 inhibitors. Two compounds, 13g and 13h, depleted the CD133+ putative cancer stem cell pool, synergized with cisplatin, and achieved efficacious concentrations in vivo following IP administration. Compound 13h additionally synergized with cisplatin in a patient-derived ovarian cancer spheroid model.