Synthesis and Chemical and Biological Comparison of Nitroxyl- and Nitric Oxide-Releasing Diazeniumdiolate-Based Aspirin Derivatives

Abstract

Structural modifications of non-steroidal anti-inflammatory drugs (NSAIDs) have successfully reduced the side effect of gastrointestinal ulceration without affecting anti-inflammatory activity, but may increase risk of myocardial infarction with chronic use. That nitroxyl (HNO) reduces platelet aggregation, preconditions against myocardial infarction and enhances contractility led us to synthesize a diazeniumdiolate-based HNO releasing aspirin and to compare it to an NO-releasing analogue. Here, the decomposition mechanisms are described for these compounds. In addition to protection against stomach ulceration, these prodrugs also exhibited significantly enhanced cytotoxcity compared to either aspirin or the parent diazeniumdiolate toward non-small cell lung carcinoma cells (A549) but were not appreciably toxic toward endothelial cells (HUVECs). The HNO-NSAID prodrug inhibited cylcooxgenase-2 and glyceraldehyde 3-phosphate dehydrogenase activity and triggered significant sarcomere shortening compared to control on murine ventricular myocytes. Together, these anti-inflammatory, anti-neoplasic and contractile properties suggest the potential of HNO-NSAIDs in the treatment of inflammation, cancer or heart failure.

Chemotherapeutic Potential of Diazeniumdiolate-Based Aspirin Prodrugs in Breast Cancer

“…IPA/NO-aspirin and DEA/NO-aspirin (Scheme 1) offer the benefits of being of the same compound class, thus facilitating comparison of the impact of respective HNO or NO donation. Furthermore, the mechanisms of decomposition have been previously elucidated [26], and it is clear that these compounds decompose intracellularly. Here, we examine in detail the impacts of these prodrugs on breast cancer progression.…”

“…IPA/NO, DEA/NO, IPA/NO-aspirin and DEA/NO-aspirin were synthesized and purified according to previously published procedures [26,86]. Stock solutions of NONO-aspirin prodrugs (100-1000×) were prepared in DMSO and stored at −20°C.…”

“…While the cytotoxicity of both aspirin and ionic diazeniumdiolates in various cancer cells lines is low (IC 50 > 1 mM) [72,83,97], NO-NSAIDs and related constructs significantly inhibit growth of cultured cancer cells as well as xenografts in mouse models [13,26,30,64,[97][98][99][100][101][102]. Keefer and colleagues have previously shown that the derivatized diazeniumdiolate JS-K was significantly cytotoxic toward a variety of breast cancer cell lines (>50% reduction in viability at 1 μM JS-K) [102].…”

“…Interestingly, the covalent linker itself has been suggested to reduce NSAID toxicity by increasing stability and absorption rate of the prodrug in the gastrointestinal tract [25]. The linker can also augment localized delivery of NO [26,27], which is a critical feature in NO donor-based treatment of conditions other than hypertension. Cleavage of the linker may also produce bioactive agents.…”

“…In addition, diazeniumdiolates can be tuned to function as HNO donors, by utilizing a primary rather than secondary amine as the scaffold [82][83][84][85]. Recently, we prepared two new diazeniumdiolate-NSAID adducts (NONO-NSAIDs) by derivatizing both a primary and secondary amine-based diazeniumdiolate with aspirin to produce [26]. A comparison of their chemical and biological properties demonstrated the expected retention of the analgesic and anti-inflammatory properties of aspirin as well as known effects of HNO or NO donors.…”