Anti-tumor Efficacy Assessment of the Sigma Receptor Pan Modulator RC-106. A Promising Therapeutic Tool for Pancreatic Cancer

Introduction: Pancreatic cancer (PC) is one of the most lethal tumor worldwide, with no prognosis improvement over the past 20-years. The silent progressive nature of this neoplasia hampers the early diagnosis, and the surgical resection of the tumor, thus chemotherapy remains the only available therapeutic option. Sigma receptors (SRs) are a class of receptors proposed as new cancer therapeutic targets due to their over-expression in tumor cells and their involvement in cancer biology. The main localization of these receptors strongly suggests their potential role in ER unfolded protein response (ER-UPR), a condition frequently occurring in several pathological settings, including cancer. Our group has recently identified RC-106, a novel pan-SR modulator with good in vitro antiproliferative activities toward a panel of different cancer cell lines. In the present study, we investigated the in vitro properties and pharmacological profile of RC-106 in PC cell lines with the aim to identify a potential lead candidate for the treatment of this tumor. Methods: Pancreatic cancer cell lines Panc-1, Capan-1, and Capan-2 have been used in all experiments. S1R and TMEM97/S2R expression in PC cell lines was quantified by Real-Time qRT-PCR and Western Blot experiments. MTS assay was used to assess the antiproliferative effect of RC-106. The apoptotic properties of RC-106 was evaluated by TUNEL and caspase activation assays. GRP78/BiP, ATF4, and CHOP was quantified to evaluate ER-UPR. Proteasome activity was investigated by a specific fluorescent-based assay. Scratch wound healing assay was used to asses RC-106 effect on cell migration. In addition, we delineated the in vivo pharmacokinetic profile and pancreas distribution of RC-106 in male CD-1 mice. Results: Panc-1, Capan-1, and Capan-2 express both SRs. RC-106 exerts an antiproliferative and pro-apoptotic effect in all examined cell lines. Cells exposure to RC-106 induces the increase of the expression of ER-UPR related proteins, and the inhibition of proteasome activity. Moreover, RC-106 is able to decrease PC cell lines motility. The in vivo results show that RC-106 is more concentrated in pancreas than plasma. Conclusion: Overall, our data evidenced that the pan-SR modulator RC-106 is an optimal candidate for in vivo studies in animal models of PC.

[1]  Yirui Hu,et al.  Bortezomib-Induced Pulmonary Toxicity: A Case Report and Review of Literature , 2018, Case reports in medicine.

[2]  E. W. Nielsen,et al.  Lactate Gap: A Diagnostic Support in Severe Metabolic Acidosis of Unknown Origin , 2018, Case reports in medicine.

[3]  A. Tesei,et al.  Sigma Receptors as Endoplasmic Reticulum Stress “Gatekeepers” and their Modulators as Emerging New Weapons in the Fight Against Cancer , 2018, Front. Pharmacol..

[4]  O. Joseph Trask,et al.  Concerns, challenges and promises of high-content analysis of 3D cellular models , 2018, Nature Reviews Drug Discovery.

[5]  B. Law,et al.  The unfolded protein response as a target for anticancer therapeutics. , 2018, Critical reviews in oncology/hematology.

[6]  B. Penke,et al.  The Role of Sigma-1 Receptor, an Intracellular Chaperone in Neurodegenerative Diseases , 2017, Current neuropharmacology.

[7]  C. Hetz,et al.  The Unfolded Protein Response and Cell Fate Control. , 2017, Molecular cell.

[8]  Alessandro Bevilacqua,et al.  ReViMS: Software tool for estimating the volumes of 3-D multicellular spheroids imaged using a light sheet fluorescence microscope. , 2017, BioTechniques.

[9]  D. Rossi,et al.  Are sigma receptor modulators a weapon against multiple sclerosis disease? , 2017, Future medicinal chemistry.

[10]  I. Katona,et al.  The ALS-linked E102Q mutation in Sigma receptor-1 leads to ER stress-mediated defects in protein homeostasis and dysregulation of RNA-binding proteins , 2017, Cell Death and Differentiation.

[11]  Michael D. Wood,et al.  Identification of the gene that codes for the σ2 receptor , 2017, Proceedings of the National Academy of Sciences.

[12]  G. Fimia,et al.  Endoplasmic Reticulum Stress, Unfolded Protein Response, and Cancer Cell Fate , 2017, Front. Oncol..

[13]  A. Tesei,et al.  Are sigma modulators an effective opportunity for cancer treatment? A patent overview (1996-2016) , 2017, Expert opinion on therapeutic patents.

[14]  D. Rossi,et al.  Gaining in pan-affinity towards sigma 1 and sigma 2 receptors. SAR studies on arylalkylamines. , 2017, Bioorganic & medicinal chemistry.

[15]  F. Kim.  Introduction to Sigma Proteins: Evolution of the Concept of Sigma Receptors. , 2017, Handbook of experimental pharmacology.

[16]  A. Tesei,et al.  Synthesis and biological evaluation of new aryl-alkyl(alkenyl)-4-benzylpiperidines, novel Sigma Receptor (SR) modulators, as potential anticancer-agents. , 2016, European journal of medicinal chemistry.

[17]  I. Ilic,et al.  Epidemiology of pancreatic cancer , 2016, World journal of gastroenterology.

[18]  D. Grimm,et al.  Reduction of TMEM97 increases NPC1 protein levels and restores cholesterol trafficking in Niemann-pick type C1 disease cells , 2016, Human molecular genetics.

[19]  E. Weledji,et al.  How Grim is Pancreatic Cancer? , 2016, Oncology reviews.

[20]  D. Rossi,et al.  Development of easy-to-use reverse-phase liquid chromatographic methods for determining PRE-084, RC-33 and RC-34 in biological matrices. The first step for in vivo analysis of sigma1 receptor agonists. , 2016, Biomedical chromatography : BMC.

[21]  D. Rossi,et al.  Toward the identification of neuroprotective agents: g-scale synthesis, pharmacokinetic evaluation and CNS distribution of (R)-RC-33, a promising SIGMA1 receptor agonist. , 2016, Future medicinal chemistry.

[22]  A. Tesei,et al.  Preclinical evidence of multiple mechanisms underlying trastuzumab resistance in gastric cancer , 2016, Oncotarget.

[23]  Alessandro Bevilacqua,et al.  3D tumor spheroid models for in vitro therapeutic screening: a systematic approach to enhance the biological relevance of data obtained , 2016, Scientific Reports.

[24]  D. Grimm,et al.  Reduction of TMEM97 increases NPC1 protein levels and restores cholesterol trafficking in Niemann-pick type C1 disease cells , 2016, Human molecular genetics.

[25]  Nita Ahuja,et al.  Early detection of pancreatic cancer. , 2015, Chinese journal of cancer research = Chung-kuo yen cheng yen chiu.

[26]  E. Zorrilla,et al.  Sigma 1 receptor regulates the oxidative stress response in primary retinal Müller glial cells via NRF2 signaling and system xc(-), the Na(+)-independent glutamate-cystine exchanger. , 2015, Free radical biology & medicine.

[27]  K. Wakabayashi,et al.  Sigma-1 receptor is involved in degradation of intranuclear inclusions in a cellular model of Huntington's disease , 2015, Neurobiology of Disease.

[28]  Teruo Hayashi.  Sigma-1 receptor: the novel intracellular target of neuropsychotherapeutic drugs. , 2015, Journal of pharmacological sciences.

[29]  K. Fukunaga,et al.  The role of SIGMAR1 gene mutation and mitochondrial dysfunction in amyotrophic lateral sclerosis. , 2015, Journal of pharmacological sciences.

[30]  Dustin J Maly,et al.  Druggable sensors of the unfolded protein response. , 2014, Nature chemical biology.

[31]  Soo-Wan Chae,et al.  Endoplasmic Reticulum Stress and Cancer , 2014, Journal of cancer prevention.

[32]  D. Rossi,et al.  Neuroprotective effects of the Sigma-1 receptor (S1R) agonist PRE-084, in a mouse model of motor neuron disease not linked to SOD1 mutation , 2014, Neurobiology of Disease.

[33]  S. Collina.  Chemical, Pharmacological, and in vitro Metabolic Stability Studies on Enantiomerically Pure RC-33 Compounds: Promising Neuroprotective Agents Acting as σ1 Receptor Agonists. , 2014 .

[34]  A. Jemal,et al.  Cancer statistics, 2014 , 2014, CA: a cancer journal for clinicians.

[35]  Teruo Hayashi,et al.  Sigma-1 Receptor Chaperone at the ER-Mitochondrion Interface Mediates the Mitochondrion-ER-Nucleus Signaling for Cellular Survival , 2013, PloS one.

[36]  D. S. St. Clair,et al.  Progesterone receptor membrane component 1/Sigma-2 receptor associates with MAP1LC3B and promotes autophagy , 2013, Autophagy.

[37]  D. Rossi,et al.  Chemical, Pharmacological, and in vitro Metabolic Stability Studies on Enantiomerically Pure RC‐33 Compounds: Promising Neuroprotective Agents Acting as σ1 Receptor Agonists , 2013, ChemMedChem.

[38]  C. Hetz,et al.  Targeting the unfolded protein response in disease , 2013, Nature Reviews Drug Discovery.

[39]  J. Hollien,et al.  The unfolded protein response in secretory cell function. , 2012, Annual review of genetics.

[40]  C. Hetz.  The unfolded protein response: controlling cell fate decisions under ER stress and beyond , 2012, Nature Reviews Molecular Cell Biology.

[41]  R. Hotchkiss,et al.  Sigma-2 ligands induce tumour cell death by multiple signalling pathways , 2012, British Journal of Cancer.

[42]  Di Chen,et al.  Bortezomib as the first proteasome inhibitor anticancer drug: current status and future perspectives. , 2011, Current cancer drug targets.

[43]  R. Austin,et al.  Endoplasmic reticulum stress and lipid dysregulation , 2011, Expert Reviews in Molecular Medicine.

[44]  R. Tomasini,et al.  Hypoxia Induced Tumor Metabolic Switch Contributes to Pancreatic Cancer Aggressiveness , 2010, Cancers.

[45]  Sanjeev Gupta,et al.  Methods for Monitoring Endoplasmic Reticulum Stress and the Unfolded Protein Response , 2010, International journal of cell biology.

[46]  H. Erfle,et al.  Identification of cholesterol-regulating genes by targeted RNAi screening. , 2009, Cell metabolism.

[47]  E. Chevet,et al.  Integrated endoplasmic reticulum stress responses in cancer. , 2007, Cancer research.

[48]  Teruo Hayashi,et al.  Sigma-1 Receptor Chaperones at the ER- Mitochondrion Interface Regulate Ca2+ Signaling and Cell Survival , 2007, Cell.

[49]  D. Amadori,et al.  Study of molecular mechanisms of pro-apoptotic activity of NCX 4040, a novel nitric oxide-releasing aspirin, in colon cancer cell lines , 2007, Journal of Translational Medicine.

[50]  G. Morgan,et al.  Bortezomib (Velcade™) in the Treatment of Multiple Myeloma , 2006, Therapeutics and clinical risk management.

[51]  M. Tatsuta,et al.  [Early detection of pancreatic cancer]. , 2006, Nihon rinsho. Japanese journal of clinical medicine.

[52]  Raymond Sawaya,et al.  The role of autophagy in cancer development and response to therapy , 2005, Nature Reviews Cancer.

[53]  Amy S. Lee.  The ER chaperone and signaling regulator GRP78/BiP as a monitor of endoplasmic reticulum stress. , 2005, Methods.

[54]  E. Gold,et al.  Epidemiology of pancreatic cancer , 1984, World Journal of Surgery.

[55]  J. Cedarbaum.  Survival , 2004 .

[56]  S. Oyadomari,et al.  Roles of CHOP/GADD153 in endoplasmic reticulum stress , 2004, Cell Death and Differentiation.

[57]  G. Skuza.  Potential antidepressant activity of sigma ligands. , 2003, Polish journal of pharmacology.

[58]  K. Mori,et al.  Activation of the ATF6, XBP1 and grp78 genes in human hepatocellular carcinoma: a possible involvement of the ER stress pathway in hepatocarcinogenesis. , 2003, Journal of Hepatology.

[59]  M. Löhr,et al.  A comprehensive characterization of pancreatic ductal carcinoma cell lines: towards the establishment of an in vitro research platform , 2003, Virchows Archiv.

[60]  T. Maurice,et al.  Neuroprotective and anti-amnesic potentials of sigma (σ) receptor ligands , 1997, Progress in Neuro-Psychopharmacology and Biological Psychiatry.

[61]  P. Frøyen,et al.  One-pot synthesis of secondary or tertiary amines from alcohols and amines via alkoxyphosphonium salts , 1995 .

[62]  Kenneth M. Johnson,et al.  Classification and nomenclature of phencyclidine and sigma receptor sites , 1987, Trends in Neurosciences.

[63]  Joseph B. Martin.  Huntington's disease , 1984, Neurology.

[64]  D. B. Vaupel,et al.  Naltrexone fails to antagonize the σ effects of PCP and SKF 10,047 in the dog , 1983 .

[65]  D. B. Vaupel,et al.  Naltrexone fails to antagonize the sigma effects of PCP and SKF 10,047 in the dog. , 1983, European journal of pharmacology.

[66]  T. Su.  Evidence for sigma opioid receptor: binding of [3H]SKF-10047 to etorphine-inaccessible sites in guinea-pig brain. , 1982, The Journal of pharmacology and experimental therapeutics.

[67]  J. Thompson,et al.  The effects of morphine- and nalorphine- like drugs in the nondependent and morphine-dependent chronic spinal dog. , 1976, The Journal of pharmacology and experimental therapeutics.