Anti-Cancer Agents in Medicinal Chemistry (v.13, #4)
Editorial (Hot Topic: Survival Signaling Through Focal Adhesion Kinase in Tumors) by Vita M. Golubovskaya (531-531).
A Small-molecule Inhibitor, 5'-O-Tritylthymidine, Targets FAK and Mdm-2 Interaction, and Blocks Breast and Colon Tumorigenesis in vivo by Vita M. Golubovskaya, Nadia L. Palma, Min Zheng, Baotran Ho, Andrew Magis, David Ostrov, William G. Cance (532-545).
Focal Adhesion Kinase (FAK) is overexpressed in many types of tumors and plays an important role in survival. Wedeveloped a novel approach, targeting FAK-protein interactions by computer modeling and screening of NCI small molecule drug database.In this report we targeted FAK and Mdm-2 protein interaction to decrease tumor growth. By macromolecular modeling we found a modelof FAK and Mdm-2 interaction and performed screening of >200,000 small molecule compounds from NCI database with drug-likecharacteristics, targeting the FAK-Mdm-2 interaction. We identified 5'-O-Tritylthymidine, called M13 compound that significantlydecreased viability in different cancer cells. M13 was docked into the pocket of FAK and Mdm-2 interaction and was directly bound tothe FAK-N terminal domain by ForteBio Octet assay. In addition, M13 compound affected FAK and Mdm-2 levels and decreasedcomplex of FAK and Mdm-2 proteins in breast and colon cancer cells. M13 re-activated p53 activity inhibited by FAK with Mdm-2promoter. M13 decreased viability, clonogenicity, increased detachment and apoptosis in a dose-dependent manner in BT474 breast andin HCT116 colon cancer cells in vitro. M13 decreased FAK, activated p53 and caspase-8 in both cell lines. In addition, M13 decreasedbreast and colon tumor growth in vivo. M13 activated p53 and decreased FAK in tumor samples consistent with decreased tumor growth.The data demonstrate a novel approach for targeting FAK and Mdm-2 protein interaction, provide a model of FAK and Mdm-2interaction, identify M13 compound targeting this interaction and decreasing tumor growth that is critical for future targeted therapeutics.
Mitoxantrone Targets the ATP-binding Site of FAK, Binds the FAK Kinase Domain and Decreases FAK, Pyk-2, c-Src, and IGF-1R In Vitro Kinase Activities by Vita M. Golubovskaya, Baotran Ho, Min Zheng, Andrew Magis, David Ostrov, William G. Cance (546-554).
Focal Adhesion Kinase (FAK) is a non-receptor kinase that is overexpressed in many types of tumors and plays a key role incell adhesion, spreading, motility, proliferation, invasion, angiogenesis, and survival. Recently, FAK has been proposed as a target forcancer therapy, and we performed computer modeling and screening of the National Cancer Institute (NCI) small molecule compoundsdatabase to target the ATP-binding site of FAK, K454. More than 140,000 small molecule compounds were docked into the crystalstructure of the kinase domain of FAK in 100 different orientations using DOCK5.1 that identified small molecule compounds, targetingthe K454 site, called A-compounds. To find the therapeutic efficacy of these compounds, we examined the effect of twenty smallmolecule compounds on cell viability by MTT assays in different cancer cell lines. One compound, A18 (1,4-bis(diethylamino)-5,8-dihydroxy anthraquinon) was a mitoxantrone derivative and significantly decreased viability in most of the cells comparable to the to thelevel of FAK kinase inhibitors TAE-226 (Novartis, Inc) and PF-573,228 (Pfizer). The A18 compound specifically blockedautophosphorylation of FAK like TAE-226 and PF-228. ForteBio Octet Binding assay demonstrated that mitoxantrone (1,4-dihydroxy-5,8-bis[2-(2-hydroxyethylamino) ethylamino] anthracene-9,10-dione directly binds the FAK-kinase domain. In addition, mitoxantronesignificantly decreased the viability of breast cancer cells in a dose-dependent manner and inhibited the kinase activity of FAK andY56/577 FAK phosphorylation at 10-20 ?M. Mitoxantrone did not affect phosphorylation of EGFR, but decreased Pyk-2, c-Src, andIGF-1R kinase activities. The data demonstrate that mitoxantrone decreases cancer viability, binds FAK-Kinase domain, inhibits itskinase activity, and also inhibits in vitro kinase activities of Pyk-2 and IGF-1R. Thus, this novel function of the mitoxantrone drug can becritical for future development of anti-cancer agents and FAK-targeted therapy research.
Upregulation of Focal Adhesion Kinase by 14-3-3? via NF?B Activation in Hepatocellular Carcinoma by Bor-Sheng Ko, Yee-Jee Jan, Tzu-Ching Chang, Shu-Man Liang, Shyh-Chang Chen, Tzu-An Liu, Yao-Ming Wu, John Wang, Jun-Yang Liou (555-562).
Focal adhesion kinase (FAK) is implicated in cancer cell survival, proliferation and migration. Expression of FAK expressionis elevated and associated with tumor progression and metastasis in various tumors, including hepatocellular carcinoma (HCC). Increased14-3-3? expression is shown to be a potential prognostic factor to predict higher risk of distant metastasis and worse overall survival inHCC. The aim of this study is to investigate whether FAK is associated or regulated by 14-3-3? to modulate tumor progression in HCC.In this study, 114 primary HCC tumors including 34 matched metastatic tumors were subjected to immunohistochemistry analysis ofFAK and 14-3-3? expression. Overexpression of FAK was significantly associated with increased risk of extrahepatic metastasis(p=0.027) and reduced 5-year overall survival rate (p=0.017). A significant correlation of FAK and 14-3-3? expression was observed inprimary tumor (p<0.001) and also metastatic tumors. Furthermore, overexpression of 14-3-3? induced FAK expression and promoteractivity which were determined by Western blotting analysis and luciferase-reporter assay. Moreover, 14-3-3? enhanced NF?B activationand increased nuclear translocation of NF?B. Results from chromatin immunoprecipitation assay revealed that 14-3-3? induced NF?Bbinding on FAK promoter region. These findings suggest that FAK expression is correlated with and upregulated by 14-3-3? viaactivation of NF?B. Target to suppress or inactivate FAK alone, or combine with 14-3-3? is thus considered as the potential therapeuticstrategy for preventing HCC tumor progression.
Cell Survival Signaling in Neuroblastoma by Michael L. Megison, Lauren A. Gillory, Elizabeth A. Beierle (563-575).
Neuroblastoma is the most common extracranial solid tumor of childhood and is responsible for over 15% of pediatric cancerdeaths. Neuroblastoma tumorigenesis and malignant transformation is driven by overexpression and dominance of cell survival pathwaysand a lack of normal cellular senescence or apoptosis. Therefore, manipulation of cell survival pathways may decrease the malignantpotential of these tumors and provide avenues for the development of novel therapeutics. This review focuses on several facets of cellsurvival pathways including protein kinases (PI3K, AKT, ALK, and FAK), transcription factors (NF-?B, MYCN and p53), and growthfactors (IGF, EGF, PDGF, and VEGF). Modulation of each of these factors decreases the growth or otherwise hinders the malignantpotential of neuroblastoma, and many therapeutics targeting these pathways are already in the clinical trial phase of development.Continued research and discovery of effective modulators of these pathways will revolutionize the treatment of neuroblastoma.
FAK and Nanog Cross Talk with p53 in Cancer Stem Cells by Vita M. Golubovskaya (576-580).
This review is focused on the role of Focal Adhesion Kinase (FAK) signaling in cancer stem cells. The recent datademonstrate the important role of FAK in cancer stem cell proliferation, differentiation, motility, and invasion. We showed recently thatthe transcription factor Nanog binds the FAK promoter and up-regulates FAK expression, and that FAK binds Nanog and phosphorylatesit. This review discusses the interaction of FAK, Nanog, Oct-3/4, and Sox-2 signaling pathways that are critical for the regulation ofcancer stem cells. The cross-linked signaling of FAK with p53 and Nanog signaling in cancer stem cell and function and targetedtherapeutics approaches are discussed.
The Role of Focal Adhesion Kinase in Lung Cancer by Grace K. Dy (581-583).
Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase implicated in carcinogenesis through its pleitropic effects on cellproliferation, survival and metastasis. This article provides a summary of the existing data implicating FAK in lung cancer.
FAK and HAS Inhibition Synergistically Decrease Colon Cancer Cell Viability and Affect Expression of Critical Genes by Melissa Heffler, Vita M. Golubovskaya, Jeffrey Conroy, Song Liu, Dan Wang, William G. Cance, Kelli B. Dunn (584-594).
Focal adhesion kinase (FAK), hyaluronan (HA), and hyaluronan synthase-3 (HAS3) have been implicated in cancer growthand progression. FAK inhibition with the small molecule inhibitor Y15 decreases colon cancer cell growth in vitro and in vivo. HAS3inhibition in colon cancer cells decreases FAK expression and activation, and exogenous HA increases FAK activation. We sought todetermine the genes affected by HAS and FAK inhibition and hypothesized that dual inhibition would synergistically inhibit viability.Y15 (FAK inhibitor) and the HAS inhibitor 4-methylumbelliferone (4-MU) decreased viability in a dose dependent manner; viability wasfurther inhibited by treatment with Y15 and 4-MU in colon cancer cells. HAS inhibited cells treated with 2?M of Y15 showedsignificantly decreased viability compared to HAS scrambled cells treated with the same dose (p<0.05) demonstrating synergisticinhibition of viability with dual FAK/HAS inhibition. Microarray analysis showed more than 2-fold up- or down-regulation of 121 genesby HAS inhibition, and 696 genes by FAK inhibition (p<0.05) and revealed 29 common genes affected by both signaling. Among thegenes affected by FAK or HAS3 inhibition were genes, playing role in apoptosis, cell cycle regulation, adhesion, transcription, heatshockand WNT pathways. Thus, FAK or HAS inhibition decreases SW620 viability and affects several similar genes, which areinvolved in the regulation of tumor survival. Dual inhibition of FAK and HAS3 decreases viability to a greater degree than with eitheragent alone, and suggests that synergistic inhibition of colon cancer cell growth can result from affecting similar genetic pathways.
Inhibiting the Interaction of cMET and IGF-1R with FAK Effectively Reduces Growth of Pancreatic Cancer Cells in vitro and in vivo by Deniz A. Ucar, Andrew T. Magis, Di-Hua He, Nicholas J. Lawrence, Said M. Sebti, Elena Kurenova, Maria Zajac-Kaye, Jianliang Zhang, Steven N. Hochwald (595-602).
Pancreatic cancer is one of the most lethal diseases with no effective treatment. Previously, we have shown that FAK isoverexpressed in pancreatic cancer and plays a key role in cancer cell survival and proliferation. FAK has been shown to interact withgrowth factor receptors including cMET and IGF-1R. As a novel therapeutic approach, we targeted the protein interaction of FAK withgrowth factor receptors to block tumor growth, alter signaling pathways and sensitize cells to chemotherapy. We have selected a smallmolecule compound (INT2-31) that decreases phosphorylation of AKT via disrupting interaction of FAK with cMET and IGF-1R. Ourresults demonstrate that interaction of a small molecule compound with FAK decreases phosphorylation of FAK Y397 while increasingFAK Y407 phosphorylation, without inhibiting the kinase activity of FAK and dramatically reduces downstream signaling to AKT. Ourlead compound, INT2-31, demonstrates significant inhibition of tumor cell growth in two orthotopic models of pancreatic cancer. Inaddition, INT2-31increases sensitivity to gemcitabine chemotherapy in a direct fresh biopsy xenograft model of pancreatic cancergrowth.
Marine Natural Products and Related Compounds as Anticancer Agents: an Overview of their Clinical Status by Karina Petit, Jean-Francois Biard (603-631).
Marine ecosystems constitute a huge reservoir of biologically active secondary metabolites. Consequently during the last pastfew decades, several marine-derived molecules have been approved for anticancer treatment or are under clinical trials. This reviewreports the present state of the art of the sixteen molecules approved or currently on the clinical pipeline for anticancer chemotherapy.The molecules are classified according to their current status in the phase (approved / phase IV / phase III / phase II / phase I) and dataare updated to April 2012.
Synthesis and Anticancer Properties of a Novel Bis-intercalator by Wei Shen, Huimin Deng, Zhiqiang Gao (632-638).
A series of naphthalene diimide (ND)-based mono-, bis-, and tris-intercalators are synthesized and evaluated for theiranticancer activities. All compounds show anticancer activities in the micromolar range. Among them the bis-intercalator is the mostpromising. Experimental results indicate that (i) target compounds intercalate DNA and (ii) the bis-intercalator with the optimal linkershows considerably more affinity to DNA than corresponding mono-and tris-intercalators. Spectroscopic measurements indicate that theND groups bind to the double-stranded DNA (ds-DNA) in a classical threading intercalation mode, while the cationic linker reinforcesthe intercalation via electrostatic interaction with ds-DNA. In vitro cytotoxicity of the bis-intercalator towards a number of cancer cells,such as C6, HeLa, and MDA-435S, is tested and compared to that of normal cells. Attractive anticancer activity is observed with the bisintercalator,which provides a new lead in the anticancer drug design strategy.
Differential Binding Preference of Methylpheophorbide a and Its Diboronated Derivatives to Albumin and Low Density Lipoproteins by Galina V. Golovina, Georgy N. Rychkov, Valentina A. Ol'shevskaya, Andrei V. Zaitsev, Valery N. Kalinin, Vladimir A. Kuzmin, Alexander A. Shtil (639-646).
The tetrapyrrolic macrocycle and the functional groups at its periphery allow for a variety of modifications aimedat multifunctional therapeutic compounds. In particular, conjugation of boron polyhedra yields dual efficacy antitumor photo/radiosensitizers. Structural optimization of these agents presumes the identification of macromolecules that bind and transport boronatedtetrapyrroles. Using spectroscopic methods we demonstrated that methylpheophorbide a forms complexes with serum albumin and lowdensity lipoproteins (LDL) whereas two diboronated derivatives, 13(2),17(3)-[di(o-carboran-1-yl)methoxycarbonyl]pheophorbide a and13(2),17(3)-[di(1-carba-closo-dodecaboran-1-yl)methoxycarbonyl]pheophorbide a, were capable of binding to LDL but not to albumin.Molecular modeling showed a mode of interaction of methylpheophorbide a with the amino acid residues in the albumin
SJSZ Glycoprotein (38 kDa) Inhibits Cell Cycle and Oxidative Stress in N-Methyl-N`- nitro-N-nitrosoguanidine-induced ICR Mice by Jin Lee, Kye-Taek Lim (647-653).
The initiation stage of liver cancer is closely related to abnormal cell proliferation as observed for other types ofcarcinogenesis. Recently, we isolated a glycoprotein from Styrax japonica Siebold et al Zuccarini (SJSZ glycoprotein), which consists ofa carbohydrate moiety (52.64%) and a protein moiety (47.36%). In this study, the antitumoric mechanism of SJSZ glycoprotein duringthe initiation stage in N-Methyl-N`-nitro-N-nitrosoguanidine (MNNG; 40 mg/kg, BW)-induced ICR was investigated. First, we evaluatedthe activities of lactate dehydrogenase (LDH), alanine aminotransferase (ALT), thiobarbituric acid-reactive substances (TBARS), andactivities of antioxidative enzymes [superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT)] in mouse liver tissueand serum. The alpha-fetoprotein (AFP), cell cycle-related factors [cyclin D1/ cyclin dependent kinase (CDK) 4], cell cycle inhibitors(CKIs; p53, p21, and p27), and proliferating cell nuclear antigen (PCNA) were then assessed using Western Blot analysis. The results ofthis analysis showed that the SJSZ glycoprotein (10 mg/kg, BW) decreased the levels of LDH, ALT, TBARS, and the expression of AFPbut it increased the activity of hepatic anti-oxidant enzymes (SOD, GPx and CAT). In addition, the SJSZ glycoprotein (10 mg/kg,BW)was shown to decrease the expression of cyclin D1/CDK4 and PCNA and increase the expression of CKIs (p53, p21, and p27). Theresults in this study indicate that the SJSZ glycoprotein displays anti-oxidative stress and anti-cell proliferation activity in MNNGinducedICR.
Antitumoral Activity of Indole-3-carbinol Cyclic tri- and Tetrameric Derivatives Mixture in Human Breast Cancer Cells: In Vitro and In Vivo Studies by Giorgio Brandi, Alessandra Fraternale, Simone Lucarini, Mirko Paiardini, Mauro De Santi, Barbara Cervasi, Maria F. Paoletti, Luca Galluzzi, Andrea Duranti, Mauro Magnani (654-662).
Indole-3-carbinol (I3C) and its oligomeric derivatives have been widely studied for their chemopreventive andchemotherapeutic properties. We have previously shown that the I3C cyclic tetrameric derivative CTet inhibits breast cancer cellproliferation in vitro and in xenotrasplanted tumor. Here we report the antitumoral activity of a mixture of tri- and tetrameric cyclic I3Cderivatives (CTr/CTet) both in vitro (MCF-7 and MDA-MB-231 breast cancer cell lines) and in vivo in a tumor xenograft model.CTr/CTet mixture avoids the low solubility drawbacks of CTet, thus favouring its solubilization, and reducing purification process, timeand costs. CTr/CTet mixture has been shown to inhibit breast cancer cell proliferation (IC50 = 1.3 and 1.6 ?g/ml in MCF-7 and MDAMB-231, respectively) inducing the G0/1 cell cycle phase accumulation. The main molecular events related to CTr/CTet activity are theoverexpression of p21, p27 and GADD45A, nuclear translocation of FOXO3a, inhibition of Akt activity and downregulation of estrogenreceptor. In vivo, the growth of xenotransplanted tumor has been inhibited and the pro-tumoral low molecular weight cyclin Edownregulation has been detected. Our data indicate that CTr/CTet is a potential anticancer combination agent for both hormoneresponsiveand triple-negative breast tumors.
Recombinant Snake Venom Cystatin Inhibits Tumor Angiogenesis in vitro and in vivo Associated with Downregulation of VEGF-A165, Flt-1 and bFGF by Qun Xie, Nanhong Tang, Rong Wan, Yuanlin Qi, Xu Lin, Jianyin Lin (663-671).
Previous studies have shown that recombinant snake venom cystatin (sv-cystatin) inhibits the invasion and metastasis of tumorcells in vitro and in vivo. The purpose of this study was to investigate the ability of recombinant sv-cystatin to inhibit tumor angiogenesisin vitro and in vivo, and the mechanisms underlying this effect. Recombinant sv-cystatin inhibited proliferation of human umbilical veinendothelial cells (HUVECs) at 100 and 200 ?g/mL after 72, 96 and 120 h. Recombinant sv-cystatin also inhibited tumor
N6-Isopentenyladenosine and its Analogue N6-Benzyladenosine Induce Cell Cycle Arrest and Apoptosis in Bladder Carcinoma T24 Cells by Sara Castiglioni, Silvana Casati, Roberta Ottria, Pierangela Ciuffreda, Jeanette A.M. Maier (672-678).
Cytokinins are phytohormones critically involved in the regulation of plant growth and development. They also affect theproliferation and differentiation of animal cells, thus representing new tools to treat diseases that involve dysfunctional cell growth and/ordifferentiation. Recently, by performing structure-function studies on human cells, we found that only N6-isopentenyladenosine and itsbenzyl analogue N6-benzyladenosine suppress the clonogenic activity and the growth of different neoplastic cells. We here broaden ourstudies on bladder carcinoma T24 cells, because, due to the high recurrence rate of bladder cancer, new active molecules are sought tocontrast the growth of this tumor. Early events induced by N6-isopentenyladenosine and N6-benzyladenosine are the alteration of T24cell morphology and the disorganization of the actin cytoskeleton. After 24 h N6-isopentenyladenosine and N6-benzyladenosine inhibitgrowth by arresting the cells in the G0/G1 phase of the cell cycle. We also show that the two compounds induce apoptosis, an eventlinked to the activation of caspase 3. Since DNA damage is a prime factor resulting in cell cycle arrest and apoptosis, it is noteworthy thatwe do not detect any genotoxic effect upon treatment of T24 cells with N6-isopentenyladenosine and N6- benzyladenosine.Because the disruption of actin filaments leads to G1 arrest and is also implicated in apoptosis, we hypothesize that cytoskeletalrearrangement might be responsible for triggering the antiproliferative and proapotpotic effects of N6-isopentenyladenosine and N6-benzyladenosine in T24 cells.