Late-stage assembly of the trimeric linear siderophore desferrioxamine B (DFOB) native to Streptomyces pilosus involves two DesD-catalyzed condensation reactions between one N-acetyl-N-hydroxy-1,5-diaminopentane (AHDP) unit and two N-succinyl-N-hydroxy-1,5-diaminopentane (SHDP) units. AHDP and SHDP are products of DesBC-catalyzed reactions of the native diamine substrate 1,5-diaminopentane (DP). The sequence of DesD-catalyzed DFOB biosynthesis was delineated by analyzing the distribution of DFOB analogues and dimeric precursors assembled by S. pilosus in medium containing 1,4-diamino-2(E)-butene (E-DBE). Seven unsaturated DFOB analogues were produced that were partially resolved by liquid chromatography (LC). Mass spectrometry (MS) measurements reported on the combination of E-DBE- and DP-derived substrates in each trimer (uDFOA1 series, 1:2; uDFOA2 series, 2:1; uDFOA3, 3:0). MS/MS fragmentation patterns reported on the absolute position of the substrate derivative at the N-acetylated terminus, the internal region, or the amine terminus of the trimer. The uDFOA1 and uDFOA2 series each comprised three constitutional isomers (binary notation (DP-derived substrate "0," E-DBE-derived substrate "1"); direction, N-acetylated-internal-amine): uDFOA1[001], uDFOA1[010], uDFOA1[100]; and uDFOA2[011], uDFOA2[110], and uDFOA2[101]. E-DBE completely replaced DP in uDFOA3[111]. Relative concentrations of the uDFOA1 series were uDFOA1[001] ≫ uDFOA1[100] > uDFOA1[010] and of the uDFOA2 series, uDFOA2[101] > uDFOA2[011] ≫ uDFOA2[110]. Dimeric compounds assembled from one N-acetylated and one N-succinylated substrate derivative were detected as trimer precursors: dDFX[00-] ≫ udDFX[10-] > udDFX[01-] (d = dimer, vacant position "-"). Relative concentrations of all species were consistent with the biosynthetic sequence: (i) SHDP activation, (ii) condensation with AHDP to form AHDP-SHDP, (iii) SHDP activation, and (iv) condensation with AHDP-SHDP to form DFOB.

Desferrioxamine B (DFOB) was discovered in the late 1950s as a hydroxamic acid metabolite of the soil bacterium Streptomyces pilosus. The exquisite affinity of DFOB for Fe(III) identified its potential for removing excess iron from patients with transfusion-dependent hemoglobin disorders. Many studies have used semisynthetic chemistry to produce DFOB adducts with new properties and broad-ranging functions. More recent approaches in chemical biology have revealed some nuances of DFOB biosynthesis and discovered new DFOB-derived drugs and radiometal imaging agents. The current and potential applications of DFOB continue to inspire a rich body of chemical biology research focused on this bacterial metabolite.

The water solubility of a natural product-inspired octadentate hydroxamic acid chelator designed to coordinate Zr(iv)-89 has been improved by using a combined microbiological-chemical approach to engineer four ether oxygen atoms into the main-chain region of a methylene-containing analogue. First, an analogue of the trimeric hydroxamic acid desferrioxamine B (DFOB) that contained three main-chain ether oxygen atoms (DFOB-O) was generated from cultures of the native DFOB-producer Streptomyces pilosus supplemented with oxybis(ethanamine) (OBEA), which competed against the native 1,5-diaminopentane (DP) substrate during DFOB assembly. This precursor-directed biosynthesis (PDB) approach generated a suite of DFOB analogues containing one (DFOB-O), two (DFOB-O) or three (DFOB-O) ether oxygen atoms, with the latter produced as the major species. Log P measurements showed DFOB-O was about 45 times more water soluble than DFOB. Second, a peptide coupling chain-extension reaction between DFOB-O and the synthetic ether-containing endo-hydroxamic acid monomer 4-((2-(2-aminoethoxy)ethyl)(hydroxy)amino)-4-oxobutanoic acid (PBH-O) gave the water soluble tetrameric hydroxamic acid DFOB-O-PBH-O as an isostere of sparingly water soluble DFOB-PBH. The complex between DFOB-O-PBH-O and Zr(iv), examined as a surrogate measure of the radiolabelling procedure, analysed by LC-MS as the protonated adduct ([M + H], m/z = 855.2; m/z = 855.3), with supporting HRMS data. The use of a microbiological system to generate a water-soluble analogue of a natural product for downstream semi-synthetic chemistry is an attractive pathway for developing new drugs and imaging agents. The improved water solubility of DFOB-O-PBH-O could facilitate the synthesis and purification of downstream products, as part of the ongoing development of ligands optimised for Zr(iv)-89 immunological PET imaging.

Parkinson's disease (PD) is a neurodegenerative disorder characterised by the death of dopaminergic neurons in the substantia nigra pars compacta (SNpc) region of the brain and formation of α-synuclein-containing intracellular inclusions. Excess intraneuronal iron in the SNpc increases reactive oxygen species (ROS), which identifies removing iron as a possible therapeutic strategy. Desferrioxamine B (DFOB, 1) is an iron chelator produced by bacteria. Its high Fe(iii) affinity, water solubility and low chronic toxicity is useful in removing iron accumulated in plasma from patients with transfusion-dependent blood disorders. Here, lipophilic analogues of DFOB with increased potential to cross the blood-brain barrier (BBB) have been prepared by conjugating ancillary compounds onto the amine terminus. The ancillary compounds included the antioxidants rac-6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (rac-trolox, rac-TLX (a truncated vitamin E variant)), R-TLX, S-TLX, methylated derivatives of 3-(6-hydroxy-2-methylchroman-2-yl)propionic acid (α-CEHC, γ-CEHC, δ-CEHC), or 4-(5-hydroxy-3-methyl-1H-pyrazol-1-yl)benzoic acid (carboxylic acid derivative of edaravone, EDA). Compounds 2-8 could have dual function in attenuating ROS by chelating Fe(iii) and via the antioxidant ancillary group. A conjugate between DFOB and an ancillary unit without antioxidant properties (3,5-dimethyladamantane-1-carboxylic acid (AdA)) was included (9). Compounds 2-9 were more lipophilic (log P -0.05 to 3.39) than DFOB (log P -2.62) and showed an average plasma protein binding 6 times greater than DFOB. The ABTS˙ radical assay indicated 2-8 had antioxidant activity ascribable to the ancillary fragment. Administration of 2 and 9 in the mouse model of PD using the neurotoxin prodrug 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), which recapitulates elevated iron of human PD, resulted in significant neuronal protection (p < 0.05; up to 89% of that in non-lesioned control animals), demonstrating the neuroprotective potential of these compounds for PD.