Activators of ATP-gated ion channels (P2X receptors) are being synthesized and investigated for cardioprotection in collaboration with Dr. Bruce Liang (University of Connecticut). MRS2339, synthesized in our lab, is a nucleotide activator of a P2X4R ion channel present in the cardiac muscle cells. We have explored the structure activity relationships of this nucleotide. Certain phosphonate derivatives are more stable to hydrolysis than the phosphate derivative MRS2339 and are being explored in vivo. MRS2339 is currently being licensed by private industry for the treatment of heart failure. The P2X ion channels mediate a number of potent and possibly important biological effects in the cardiovascular, inflammatory, and central nervous systems. Previous studies have shown that extracellular ATP can cause an ionic current in murine, rat and guinea pig cardiac ventricular myocytes. The receptor that mediates this current appears to be a P2X receptor, of which the P2X4 receptor is an important subunit. Activation of P2X receptors leads to the opening of a nonselective cation channel permeable to sodium, potassium, and calcium ions. The current is inward at negative membrane potentials, reverses near 0 mV, and becomes outward at positive potentials. The continuous activation of this receptor channel by endogenous extracellular ATP may assume an important biological function. This constant activation under the resting or negative membrane potentials would produce an inward current, whereas its activation during depolarized portions of the action potential should lead to an outward current. These currents represent a possible ionic mechanism by which the cardiac P2X channel achieves its biological effects. A potential biologically important role of the cardiac P2X receptor was suggested by the finding that cardiac myocyte-specific overexpression of the P2X4 receptor can rescue the hypertrophic and heart failure phenotype of the calsequestrin (CSQ) model of cardiomyopathy. However, little is known regarding regulation of the cardiac P2X receptor in cardiac hypertrophy or failure. Furthermore, it is not clear whether an increased activation of the endogenous P2X receptor channel is beneficial or harmful in the progression of heart failure. The regulation of the P2X receptor-mediated ionic current and its potential role in heart failure was investigated using several novel nucleotide agonists. Chronic administration of a novel nucleotidase-resistant P2 receptor agonist MRS2339, which was capable of inducing this ionic current and was devoid of any vasodilator action, reduced cardiac hypertrophy and increased lifespan. The data suggests that an important biological function of the cardiac P2X current is to favorably modulate the progression of cardiac hypertrophy and failure. Recently we identified uncharged carbocyclic nucleotide analogues (including nonhydrolyzable phosphonates) related structurally to MRS2339, that represent potential candidates for the treatment of heart failure, suggesting this as a viable and structurally broad approach. We also found a beneficial therapeutic effect of 2-cyclohexylthio-adenosine 5-monophosphate in mice with heart failure (HF). We have provided compounds for the study of transient receptor potential cation channel subfamily V member 1 (TRPV1) to our collaborator Dr. Michael Iadarola of NIH. TRPV1 is a high-conductance, nonselective cation channel strongly expressed in nociceptive primary afferent neurons of the peripheral nervous system. In functions as a multimodal nociceptor gated by temperatures greater than 43˚C, protons, and small molecule vanilloid ligands such as capsaicin. The ability to respond to a variety of stimuli (heat, low pH, vanilloids, and endovanilloids) and its altered sensitivity and expression in experimental inflammatory and neuropathic pain models made TRPV1 a major target for the development of novel, nonopioid analgesics. These have been mostly antagonists, which have intolerable side effects in human clinical trials, but recent work shows that potent agonists or enhancers agonists have utility in this context. Here we show that the dihydropyridine derivative 4,5-diethyl-3-(2-methoxyethylthio)-2-methyl-6-phenyl-1,4-dihydropyridine-3,5-dicarboxylate (MRS1477) behaves as a positive allosteric modulator of both proton and vanilloid activation of TRPV1. Under inflammatory mimetic conditions of low pH (6.0) and protein kinase C phosphorylation, addition of MRS1477 further increased sensitivity of already sensitized TPRV1 toward capsaicin. MRS1477 does not affect inhibition by known vanilloid antagonists and remains effective in potentiating activation by pH in the presence of an orthosteric vanilloid antagonist. These results indicate a distinct site on TRPV1 for positive allosteric modulation that may bind endogenous compounds or novel pharmacological agents. Positive modulation of TRPV1 sensitivity suggests that it may be possible to produce a selective analgesia through calcium overload restricted to highly active nociceptive nerve endings at sites of tissue damage and inflammation.

Project Start
Project End
Budget Start
Budget End
Support Year
7
Fiscal Year
2013
Total Cost
$67,544
Indirect Cost
City
State
Country
Zip Code
Janowsky, Aaron; Tosh, Dilip K; Eshleman, Amy J et al. (2016) Rigid Adenine Nucleoside Derivatives as Novel Modulators of the Human Sodium Symporters for Dopamine and Norepinephrine. J Pharmacol Exp Ther 357:24-35
Jacobson, Kenneth A; Müller, Christa E (2016) Medicinal chemistry of adenosine, P2Y and P2X receptors. Neuropharmacology 104:31-49
Yang, Ronghua; Beqiri, Dardan; Shen, Jian-Bing et al. (2015) P2X4 receptor-eNOS signaling pathway in cardiac myocytes as a novel protective mechanism in heart failure. Comput Struct Biotechnol J 13:1-7
Zhou, Siyuan; Yang, Tiehong; Jacobson, Kenneth A et al. (2013) The therapeutic effect of 2-cyclohexylthio-AMP in heart failure. J Cardiovasc Pharmacol 61:553-9
Kumar, T Santhosh; Yang, Tiehong; Mishra, Shilpi et al. (2013) 5'-Phosphate and 5'-phosphonate ester derivatives of (N)-methanocarba adenosine with in vivo cardioprotective activity. J Med Chem 56:902-14
Aydemir, Tolunay Beker; Chang, Shou-Mei; Guthrie, Gregory J et al. (2012) Zinc transporter ZIP14 functions in hepatic zinc, iron and glucose homeostasis during the innate immune response (endotoxemia). PLoS One 7:e48679
Kaszas, Krisztian; Keller, Jason M; Coddou, Claudio et al. (2012) Small molecule positive allosteric modulation of TRPV1 activation by vanilloids and acidic pH. J Pharmacol Exp Ther 340:152-60
Zhou, Si-Yuan; Mamdani, Mohammed; Qanud, Khaled et al. (2010) Treatment of heart failure by a methanocarba derivative of adenosine monophosphate: implication for a role of cardiac purinergic P2X receptors. J Pharmacol Exp Ther 333:920-8
Hu, Huiling; Lu, Wennan; Zhang, Mei et al. (2010) Stimulation of the P2X7 receptor kills rat retinal ganglion cells in vivo. Exp Eye Res 91:425-32
Diaz, Juan Carlos; Simakova, Olga; Jacobson, Kenneth A et al. (2009) Small molecule blockers of the Alzheimer Abeta calcium channel potently protect neurons from Abeta cytotoxicity. Proc Natl Acad Sci U S A 106:3348-53

Showing the most recent 10 out of 13 publications