title: Novel double prodrugs of the iron chelator N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED): Synthesis, characterization, and investigation of activation by chemical hydrolysis and oxidation.
abstract: The development of iron chelators suitable for the chronic treatment of diseases where iron accumulation and subsequent oxidative stress are implicated in disease pathogenesis is an active area of research. The clinical use of the strong chelator N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED) and its alkyl ester prodrugs has been hindered by poor oral bioavailability and lack of conversion to the parent chelator, respectively. Here, we present novel double prodrugs of HBED that have the carboxylate and phenolate donors of HBED masked with carboxylate esters and boronic acids/esters, respectively. These double prodrugs were successfully synthesized as free bases (7a-f) or as dimesylate salts (8a-c,e), and were characterized by (1)H, (13)C, and (11)B NMR; MP; MS; and elemental analysis. The crystal structure of 8a was solved. Three of the double prodrugs (8a-c) were selected for further investigation into their abilities to convert to HBED by stepwise hydrolysis and H2O2 oxidation. The serial hydrolysis of the pinacol and methyl esters of N,N'-bis(2-boronic acid pinacol ester benzyl)ethylenediamine-N,N'-diacetic acid methyl ester dimesylate (8a) was verified by LC-MS. The macro half-lives for the hydrolyses of 8a-c, measured by UV, ranged from 3.8 to 26.3 h at 37 °C in pH 7.5 phosphate buffer containing 50% MeOH. 9, the product of hydrolysis of 8a-c and the intermediate in the conversion pathway, showed little-to-no affinity for iron or copper in UV competition experiments. 9 underwent a serial oxidative deboronation by H2O2 in N-methylmorpholine buffer to generate HBED (k = 10.3 M(-1) min(-1)). The requirement of this second step, oxidation, before conversion to the active chelator is complete may confer site specificity when only localized iron chelation is needed. Overall, these results provide proof of principle for the activation of the double prodrugs by chemical hydrolysis and H2O2 oxidation, and merit further investigation into the protective capabilities of the prodrugs against H2O2-induced cell death.
authors.affiliation: Department of Medicinal Chemistry, University of Florida, Gainesville, FL 32610, USA. Electronic address: nikkithiele@yahoo.com.
authors.forename: Nikki A
authors.initials: NA
authors.lastname: Thiele
authors.affiliation: Department of Chemistry, University of Florida, Gainesville, FL 32611, USA. Electronic address: abboud@chem.ufl.edu.
authors.forename: Khalil A
authors.initials: KA
authors.lastname: Abboud
authors.affiliation: Department of Medicinal Chemistry, University of Florida, Gainesville, FL 32610, USA. Electronic address: sloan@cop.ufl.edu.
authors.forename: Kenneth B
authors.initials: KB
authors.lastname: Sloan
mesh_terms: D001897:Boronic Acids / Q000737:chemistry; D002463:Cell Membrane Permeability; D003300:Copper / Q000737:chemistry; D004492:Edetic Acid / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D006868:Hydrolysis; D007501:Iron / Q000737:chemistry; D007502:Iron Chelating Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D011355:Prodrugs / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism*; D012995:Solubility; D014867:Water / Q000737:chemistry
chemical_list: D001897:Boronic Acids; D007502:Iron Chelating Agents; D011355:Prodrugs; D014867:Water; C037740:N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid; D003300:Copper; D004492:Edetic Acid; D007501:Iron
keywords: Boronic acid; Chelation therapy; Double prodrug; Iron; Oxidative stress; Prochelator