Abstract
Iron levels in mitochondria are critically important for the normal functioning of the organelle. Abnormal levels of iron and the associated formation of toxic oxygen radicals have been linked to a wide range of diseases and consequently it is important to be able to both monitor and control levels of the mitochondrial labile iron pool. To this end a series of iron chelators which are targeted to mitochondria have been designed. This overview describes the synthesis of some of these molecules and their application in monitoring mitochondrial labile iron pools and in selectively removing excess iron from mitochondria.
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Introduction
Iron represents a vital element required for almost all forms of life. Because of its ability to transfer electrons by shuttling between ferrous and ferric forms, it is a crucial component participating in the electron transport chain within mitochondria. In biological systems, iron can be incorporated into iron–sulfur [Fe–S] clusters and the heme molecule (Paul and Lill 2015; Ponka 1999). Both of these iron-containing structures serve as cofactors for many enzymes and are partially synthesized inside mitochondria, rendering the mitochondrion the central organelle in cellular iron metabolism. This critical dependence of mitochondria on iron renders the specific quantification of the mitochondrial labile iron pool in both normal and disease states highly desirable (Urrutia et al. 2014). Furthermore, as labile iron is redox active, its presence will contribute to the production of toxic reactive oxygen species—(ROS). Thus, in some disease states it will be an advantage to have the capability of controlling levels of mitochondrial labile iron (Reelfs et al. 2016; Sandoval-Acuna et al. 2021).
Neurodegeneration
Neurodegenerative diseases are a heterogeneous group of disorders characterized by gradually progressive, selective loss of anatomically or physiologically related neuronal systems. Prototypical examples include Alzheimer’s disease (AD), Parkinson’s disease (PD). It is clear that mitochondrial involvement is an important common theme in both of these disease states. Mitochondria are key regulators of cell survival and death and have a central role in ageing (Lin and Beal 2006). There is strong evidence that mitochondrial dysfunction occurs early and acts causally in the disease pathogenesis. Furthermore, it is clear that many neurodegenerative diseases are associated with accumulation of iron in the central nervous system (Ke and Qian 2003) and it will be important to establish the changes in mitochondrial labile iron levels between normality and the diseased state.
Pathological levels of iron are found in association with aggregated amyloid-β protein (A β) in the extracellular space of AD patients. There is evidence that iron accumulation in affected regions of the AD brain may outstrip the ability of ferritin to safely sequester it, which would lead to pathological ROS formation through Fenton chemistry (Connor et al. 1992). Indeed, oxidative stress is a well-documented feature of AD and there is evidence to support the concept that iron contributes to oxidative stress in AD (Altamura and Muckenthaler 2009). Clearly there is an urgency to establish the labile iron levels in Alzheimer brain mitochondria.
Iron accumulation in substantia nigra has been a recognised feature of Parkinson’s disease (PD) (Dexter et al. 1987). Excessive cellular iron, through its propensity to generate ROS, has consequences for many of the well-established pathogenic mechanisms of PD. Mitochondrial dysfunction was one of the earliest features identified in the study of subcellular pathology in PD. Reduced activity of mitochondrial complex I has been reported in the brain, platelets, and skeletal muscle of patients with idiopathic PD (Krig et al. 1992). A number of chelators have been directed at the removal of iron from the brains of PD patients, the most extensively studied being deferiprone (Devos et al. 2014). A disadvantage of deferiprone is that it lacks a controlled cellular distribution.
Cardiovascular disease
It is well established that many cardiovascular disease risk factors are associated with mitochondrial dysfunction, for instance atherosclerosis, hypercholesterolemia, diabetes, ischemia–reperfusion injury and exposure to tobacco smoke. Indeed, the iron chelator deferiprone is capable of removing iron from mitochondria and reversing the symptoms of chronic obstructive pulmonary disease in mice (Cloonan et al. 2016). The disadvantage of using deferiprone, as indicated above, is that it is not targeted to the mitochondria.
Skin photodamage
The presence of appreciable levels of labile iron in the mitochondria of fibroblasts and keratinocytes render these cells sensitive towards oxidative damage on exposure to ultraviolet A (UVA, 320–400 nm, the oxidising component of sunlight). The harmful consequences of iron-catalysed damage exerted by UVA have been shown to play a key role in skin photoaging and photocarcinogenesis (Dissemond et al. 2003). In this regard, solar UVA is recognised as a strong membrane damaging agent promoting lipid peroxidation in subcellular organelle membranes of skin fibroblasts and keratinocytes via pathways involving singlet oxygen and labile iron (Vile and Tyrrell 1995). Furthermore, the labile iron-mediated oxidative damage caused by UVA to mitochondrial membranes has been shown to cause necrotic cell death via ATP depletion (Zhong et al. 2004; Aroun et al. 2012).
Mitochondrial-targeted chelators
Because of the crucial role of mitochondrial iron overload in a growing number of oxidative conditions and pathologies, there is a clear need to design mitochondrial iron probes with high specificity and sensitivity to evaluate the labile iron of the organelles (Rouault 2016; Gao et al. 2021; Mena et al. 2015). The criteria for the design of mitochondria-specific iron sensors imply that iron sensors reside exclusively in the organelles and provide a reliable quantification of mitochondrial labile iron. In this regard, we have developed a series of highly specific fluorescent mitochondria-targeted iron sensors that fulfil the above criteria as they can reach mitochondria by means of mitochondria-homing peptide sequences (Abbate et al. 2015, 2016). This successful design was initially reported by Schiller et al. (2000) and Zhao et al. (2003), whereby tetrapeptides with alternating aromatic and basic amino acids were demonstrated to penetrate cell membranes and to concentrate in mitochondria. By incorporating D-amino acids in the C-amidated peptides, resistance to peptidase attack is achieved. Although a wide range of arginine containing peptides have been reported to be able to permeate membranes and to concentrate in the mitochondria (Futaki 2006; Horton et al. 2008; Nakose et al. 2012), the SS-like peptides (Schiller and Szeto peptides) possess the advantage of being tetrapeptides (Zhao et al. 2003) and therefore they lack a pronounced secondary structure; a clear advantage for drug design. By adopting this alternating aromatic/basic residue peptide design, Horton et al (2008) and Abbate et al. (2015, 2016) have prepared fluorescent mitochondria-targeted peptides (e.g. BP19) which are capable of being selectively accumulated by mitochondria. The net positive charge of these peptide derivatives ensures that they are accumulated by mitochondria due to the large membrane potential of the inner mitochondrial membrane. Furthermore, by attaching an iron-chelating residue (Fig. 1), such peptides are capable of evaluating the mitochondrial labile iron pool (Abbate et al. 2016). Leading on from these findings, Reelfs et al. using hexadentate iron chelators, covalently attached to mitochondrial-targeted peptides (e.g. BP29, Fig. 1), have demonstrated that such molecules can selectively scavenge excess iron from mitochondria with beneficial effects (Reelfs et al. 2016).
In parallel with the above studies, Espósito et al. (Alta et al. 2017a) have reported triphenylphosphonium-deferoxamine as a candidate mitochondrial targeted iron chelator. They demonstrate that this derivatised siderophore can penetrate plasma membranes and is clearly accumulated in mitochondria. More recently, triphenyl phosphonium-deferoxamine has been demonstrated to suppress tumour growth and metastasis by scavenging mitochondrial labile iron (Sandoval-Acuña et al. 2021). However, a potential toxicity issue with the application of the triphenyl phosphonium moiety is that it can induce non-specific effects due to the accumulation of the extremely lipophilic TPP+ cation into membranes (Smith et al. 2003; Murphy, 2008). This issue is avoided by the utilisation of readily biodegradable peptides, such as the SS-like peptides which are reported in the present study. Significantly, Espósito et al. have also reported the application of basic/amphiphilic peptide address systems for the targeting of deferoxamine to mitochondria (Alta et al. 2017b).
Another interesting mitochondrial directed iron(II) probe is SiRhoNox-1 (mito-Ferro Green) (Hirayama et al. 2013, 2017, 2019), although its precise mode of interaction with Fe2+ ions still remains to be confirmed. The Fe2+—mediated deoxygenation of the N-oxide group on the fluorophore leads to an enhanced fluorescence. The sensitivity towards Fe2+ is limited to approximately 1 μM and the reduction of the N-oxide may also be facilitated by reaction with glutathione, a thiol that reaches high mM concentrations in the mitochondria. Consequently, it would appear that this probe should be subjected to more carefully controlled studies, before it can be recognised as a truly specific mitochondrial iron(II) probe.
Results
Synthesis of BP19 (1)
Solid-phase peptide synthesis (SPPS)
Abbate et al. recently reported the synthesis of BP19 (1) using a solid-phase peptide synthesis (SPPS) approach (Abbate et al. 2015). SPPS is generally the first method of choice for the chemical synthesis of peptides. The derivatised mimosine analogue (Scheme 1d) was attached to a solid-phase resin as indicated in Scheme 1. A two stage deprotection (Scheme 1e and f) led to the isolation of crude BP19 as an amorphous white solid (80% yield). Further purification was achieved by preparative high-pressure liquid chromatography (HPLC).
Solution synthesis
Unfortunately, although SPPS can be automated and is scalable, it suffers from a negative environmental footprint mainly due to extensive solvent use and low yields (Martin et al. 2020). In line with the modern effort of academia and industry to render peptide synthesis greener and to optimize the yields, we here report an alternative solution-based synthetic method for BP19.
As reported in Fig. 2, our strategy was to divide the final structure into three building blocks (BBs) choosing a convergent synthetic approach over a sequential approach in order to achieve higher yields. These three BBs were synthesized singularly as reported in Schemes 2, 3, and 4 and were then used for the final assembly (Scheme 5) of the product BP19 (1).
The synthesis of BB1 (Scheme 2) was designed as a two-step reaction starting from commercially available Boc-l-Phe-OH that was first activated with DCC/NHS to produce the NHS derivative 3 and then coupled with d-Arg(Pbf)-OH (4) to produce the first BB (5). The synthesis of BB2 (Scheme 3) was achieved with one step reaction starting from Boc-l-Orn-OH (6) and 3-(benzyloxy)-2-methyl-4H-pyran-4-one (7). Molecule 7 was produced as previously reported (Cilibrizzi et al. 2018) and then reacted as classically reported with the amino group of 6 converting the pyran-4-one ring of 7 in the derivative N-alkylpyridones 8 (BB2). BB3 was synthesized in four stepwise reactions (Scheme 4) starting from Z-l-Dap(Boc)-OH (9). Molecule 9 was firstly converted into its amide derivative 10. The reaction was initially tried with ammonia under different conditions (1. Et3N, ClCOOEt, THF, -5 °C to rt in 2 h and 2. EDC, HOBt, Et3N, DMF, rt in 24 h), but higher yields were achieved when the reaction was conducted as reported by Talukdar et al (2005) with Boc2O, ammonium bicarbonate and pyridine in acetonitrile at room temperature for 16 h. The resulting amide (10) was then deprotected at its β amino group with TFA and the deprotected compound 11 was coupled with dansyl chloride (12) giving compound 13. BB3 (14) was finally obtained by deprotection of the amino group of 13 with 10% palladium on carbon under an atmosphere of H2 for 12 h.
The final assembly (Scheme 5) of BP19 (1) started from the coupling of BB2 (8) and BB3 (14). The reaction was achieved using EDC/HOBt as coupling reagents to give the product 15 which was subsequently deprotected with TFA giving intermediate 16. BB1 (5) was then coupled with molecule 16 with EDC/HOBt giving the protected target molecule 17. Treatment of molecule 17 with neat TFA for 24 h facilitated the simultaneous deprotection of the labile -Boc and -Pbf protecting groups and the -Bn group on the hydroxypyridinone ring giving the final molecule 1 (BP19).
Synthesis of BP29 (2)
BP29 was synthesised using SPPS in similar fashion to that of BP19 (Scheme 6). Each of the three lysine residues on the resin-bound peptide were deprotected and then conjugated to O-dimethyl-2,3-dihydroxybenzoic acid. Final deprotection led to the production of crude BP29 as a white amorphous solid (80% yield). This product was purified by preparative HPLC.
Properties of the mitochondria-targeted iron sensor BP19
BP19 was selected from a range of probes for the selective measurement of labile iron in the mitochondria. It was demonstrated that BP19 was preferentially accumulated in the mitochondria of FEK4 cells (human primary foreskin fibroblasts) Fig. 3 (Abbate et al. 2015). Furthermore, when the cells were loaded with iron the fluorescence signal was reduced (Fig. 4) and when the added iron was chelated by deferiprone, the fluorescence was reinstated.
BP19 has been utilised to evaluate the level of mitochondrial labile iron of cultured fibroblasts obtained from Friedreich’s ataxia (FRDA) patients when compared to skin fibroblasts from healthy donors. This study revealed that the mean levels of mitochondrial labile iron in FRDA fibroblasts were on average sixfold higher than those in healthy fibroblasts (Fig. 5) (Reelfs et al. 2019a). The mean levels of mitochondrial labile iron in FRDA cells were 1.11 ± 0.37 μM, whereas those from healthy donors were 0.17 ± 0.12 μM. While it is known that in the neuromuscular disorder FRDA, the decreased iron-sulphur cluster and heme formation by defective frataxin protein causes the pathological accumulation of redox-active labile iron in mitochondrial compartments (Rouault 2016; Llorens et al. 2019; Chiang et al. 2020), our study was the first to provide an estimate of the extent of mitochondrial iron overload in cells derived from FRDA patients.
Cellular studies with mitochondrial-targeting iron chelators
In an attempt to increase the potency of iron chelation by the use of mitochondria targeted peptides, hexadentate analogues designed in a similar fashion to siderophore structures (Hider and Kong 2010), were investigated (Reelfs et al. 2016). In this context, we synthesized a series of chimeric hexapeptides, containing 3 iron chelating residues attached to a SS-like peptide. The iron-chelating moieties were of either catechol e.g. BP29 (Fig. 1) or hydroxypyridinone types. Depending on the application, both types of hexapeptide have demonstrated significant potency in various investigations.
The cytoprotective potential of BP29 against UVA-induced damage in fibroblasts
We reasoned that the use of mitochondria-targeted iron chelators to specifically remove the mitochondrial labile iron may be an effective approach to protect the skin cells against the harmful effects of UVA. In this regard, we demonstrated that pre-treatment of human primary skin fibroblasts with the mitochondria-targeted tri-catechol-based iron chelator linked to mitochondria-homing SS-peptides (BP29) exhibits an unprecedented protection against UVA-induced oxidative damage to mitochondrial membrane and the ensuing ATP depletion and necrotic cell death (Fig. 6) (Reelfs et al. 2016). The remarkable potency of this mitochondria-targeted chelator peptide was dependent on its effective uptake by skin cells and its ability to selectively partition to mitochondria, thereby removing the excess of potentially harmful labile iron of the organelle. To our knowledge this is the first mitochondria-targeted iron chelator of its kind to demonstrate promising potential for skin photoprotection against the deleterious effects of UVA component of sunlight.
The cytoprotective potential of BP29 against UVA-induced damage in FRDA fibroblasts
We have recently reported that FRDA skin fibroblasts are highly susceptible to UVA-induced oxidative damage and cell death when compared to healthy control skin fibroblasts (see Reelfs et al. 2019a and Fig. 7). Pre-treatment of FRDA fibroblasts with BP29 was found to rescue the cells against a high but physiologically-relevant dose of UVA (i.e. 500 kJ/m2 equivalent to 3.5 h in sunlight at sea level) (Fig. 7) (Reelfs et al. 2019a). This study highlighted the potential of mitochondria-targeted iron chelators for efficient skin photoprotection in FRDA patients.
The cytoprotective potential of BP29 against H2O2-induced toxicity in FRDA fibroblasts
The higher mitochondrial labile iron of skin fibroblasts from FRDA patients renders them highly susceptible to iron stress and to be appreciably more sensitive to H2O2-mediated cell death than controls, in line with the importance of the organelles’ labile iron under oxidative stress conditions and pathologies (Wong et al. 1999; Lim et al. 2008; Pourzand et al; 2019). Iron chelation therapy of FRDA patients has demonstrated that chelators such as deferiprone can provide significant and measurable impact on patients’ neurological functions (Boddaert et al. 2007). Nevertheless such chelators lack intracellular organelle specificity. In order to establish whether mitochondrial targeting enhances the efficacy of iron chelators, we compared the effect of BP29 with two clinically used iron chelators desferrioxamine and deferiprone. Fibroblasts derived from healthy donors and FRDA patients were pre-treated with iron chelators and then challenged with the final H2O2 concentration of 100 μM (for 1 h). The cytotoxicity tests performed 24 h after the H2O2 treatment revealed that BP29 was just as effective as desferrioxamine and deferiprone at preventing oxidative stress in both cell lines (Fig. 8). (Pourzand et al. 2019).
The neuroprotective potential of BP29-type molecules against oxidative injuries in Parkinson’s disease
Mitochondrial dysfunction in PD results in detrimental mitochondrial iron overload accompanied with an increase of chelatable redox-active labile iron and consequent excess of labile iron-driven production of harmful reactive oxygen species in specific regions of the nervous system (Devos et al. 2014; Urrutia et al. 2021; Deus et al. 2021). Dopaminergic neuron degeneration follows as a consequence of oxidative stress. This suggests that targeting the organelle’s excess labile iron for removal may be an effective approach for the successful therapy of PD. In order to investigate this possibility, the cytoprotective potential of a BP29-type hexadentate mitochondria-targeted iron chelator (PD2) was evaluated in SH-SY5Y cells, an in vitro model of PD, against 6-hydroxydopamine (6-OHDA)-induced PD-like mitochondrial dysfunction (Fig. 9). Our results demonstrated that the SH-SY5Y cell line, when pre-treated with compound PD2, exhibited the highest protection by significantly protecting the cells up to 40% against 6-OHDA-induced necrotic cell death. Our results further revealed that compound PD2 afforded up to 80% protection against mitochondria membrane depolarization which accompanies 6-OHDA-induced cell death. The cytoprotective properties displayed in vitro by PD2 demonstrate the validity of such compounds to protect cells against the consequences of mitochondrial dysfunction such as that which are seen in the brain cells from patients with PD (Reelfs et al. 2019b).
Conclusions
Directing iron-selective chelators to the mitochondria offers a means of chelating mitochondrial excess iron with minimal iron scavenging activity in the cytosol, nucleus and lysosome. In principle this should reduce the toxicity of such chelators. In this study we describe the synthesis of one such molecule. We have demonstrated that such chelators, modelled on siderophore structure, have cytoprotective potential in a range of cell types.
It will be important to be able to measure the labile iron levels in mitochondria and to this end we report the synthesis and design of a mitochondrial targeted iron selective fluorescent probe. Using this probe, we have demonstrated a large increase in the mitochondrial labile iron pool of fibroblasts isolated from FRDA patients.
Synthetic details
Materials and chemicals were purchased from Acros Organics, Merck, Fisher Scientific International, and Fluorochem Ltd and were reagent grade or better. Solvents and NMR solvents were purchased from Fisher Scientific, Merck KGaA, and VWR. Silica gel for column chromatography was purchased from Merck. All samples were dried in a vacuum oven connected to a vacuum pump (BOC-Edwards.). Silica gel 60 F254, Merck pre-coated aluminum sheets were employed for thin-layer chromatography (TLC) and spots were visualized under UV light. 1H NMR and 13C NMR spectra were recorded on A Bruker Avance III HD NanoBay 400 MHz NMR with a 5 mm 1H/13C/15N/31P QNP probe equipped with z-gradient. Tetramethylsilane (TMS) was used in all NMR experiments as internal standard and chemical shift (δ) values are given in ppm. Low resolution mass spectra were obtained on a Thermofisher LCQ DECA XP ion trap mass spectrometer or a Waters—Micromass ZQ—Single quadrupole mass spectrometer. High-resolution Mass Spectrometry (HRMS) was conducted on a Thermo Fisher Scientific Exactive Mass Spectrometer operating in positive electrospray ionisation mode.
3 (2,5-dioxopyrrolidin-1-yl (tert-butoxycarbonyl)-l-phenylalaninate). N, N′-Dicyclohexylcarbodiimide (DCC) (1.01 g, 4.90 mmol, 1.3 eq) and N-hydroxysuccinimide (0.56 g, 4.90 mmol, 1.3 eq) were added to a solution of Boc-l-Phe-OH (2) (1.00 g, 3.76 mmol, 1.0 eq) dissolved in 50 mL of dimethylformamide (DMF) and stirred for 3 h at room temperature. The mixture was filtered, the filtrate was concentrated under reduced pressure and the obtained compound 3 was used without any further purification. Yield 99%.
5 (N2-((tert-butoxycarbonyl)-l-phenylalanyl)-Nw-((2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-5-yl)sulfonyl)-d-arginine). Compound 3 (0.93 g, 2.57 mmol, 1.1 eq) was added to a solution of d-Arg(Pbf)-OH (4) (1.00 g, 2.34 mmol, 1.0 eq) and triethylamine (1.30 mL, 9.37 mmol, 4.0 eq) in DMF (50 mL). The reaction was stirred at room temperature for 24 h. The solvent was removed under reduced pressure and the crude product was purified by column chromatography using a gradient of EtOH/NH4OH/DCM/Pet. Et. from13.7:1.7:71.9:12.7 to 23.5:4.7:62.5:9.4 as eluents. Yield 75%. 1H NMR (400 MHz, Methanol-d4) δ 7.22 (d, J = 6.1 Hz, 5H), 7.19–7.11 (m, 1H), 4.29 (d, J = 6.1 Hz, 1H), 4.23 (s, 1H), 3.11 (dd, J = 13.7, 5.5 Hz, 3H), 2.99 (s, 2H), 2.82 (dd, J = 13.7, 8.8 Hz, 1H), 2.58 (s, 3H), 2.52 (s, 3H), 2.08 (s, 3H), 1.75 (s, 1H), 1.56 (dt, J = 13.7, 7.5 Hz, 1H), 1.45–1.29 (m, 15H). 13C NMR (101 MHz, Methanol-d4) δ 173.63, 159.84, 157.52, 139.40, 138.65, 134.39, 133.49, 130.45, 130.35, 129.47, 129.26, 127.72, 127.52, 126.02, 118.44, 87.64, 80.70, 57.77, 54.45, 43.97, 39.39, 30.56, 28.74, 28.70, 28.66, 26.44, 19.65, 18.43, 12.56. HRMS Calculated [M + H]: 674.3218; Found [M + H]; 674.3208.
8 ((S)-5-(3-(benzyloxy)-2-methyl-4-oxopyridin-1(4H)-yl)-2-((tert-butoxycarbonyl)amino)pentanoic acid). 3-(Benzyloxy)-2-methyl-4H-pyran-4-one (7) (1.1 g, 4.64 mmol, 1.1 eq) was added to a stirred solution of Boc-l-Orn-OH (6) (980 mg, 4.22 mmol, 1.0 eq) in ethanol/water 1:1 (20 mL) and the pH was adjusted to 10.5 using 1 N sodium hydroxide solution and the mixture refluxed for 24 h. The solvent was evaporated under vacuum. The reaction was acidified with 6 N HCl until pH 7 and extracted with DCM (3 × 40 mL). The organic layers were dried over anhydrous sodium sulfate, filtered, and rotary evaporated to give an orange oil. Further purification was obtained by flash column chromatography using a gradient of DCM/MeOH from 8:2 to 6:4 as eluents. Yield 32%. 1H NMR (400 MHz, Methanol-d4) δ 7.71 (d, J = 7.4 Hz, 1H), 7.35 (ddd, J = 11.3, 6.4, 3.2 Hz, 5H), 6.49 (d, J = 7.4 Hz, 1H), 5.07 (s, 2H), 4.00 (tt, J = 14.2, 6.4 Hz, 3H), 2.15 (s, 3H), 1.89–1.54 (m, 4H), 1.44 (s, 9H). 13C NMR (101 MHz, Methanol-d4) δ 174.58, 158.07, 145.28, 141.24, 138.35, 130.35, 129.46, 117.34, 80.52, 74.57, 54.80, 29.99, 28.78, 28.09, 12.85. HRMS Calculated [M + H]: 431.2711; Found [M + H]; 431.2171.
10 (benzyl tert-butyl (3-amino-3-oxopropane-1,2-diyl)(S)-dicarbamate). To Z-l-Dap(Boc)-OH (9) (1.00 g, 2.95 mmol, 1.0 eq) in dry acetonitrile (40 ml), Boc2O (1.29 g, 5.91 mmol, 2.0 eq), ammonium bicarbonate (467 mg, 5.91 mmol, 2.0 eq) and pyridine (0.88 mL, 10.91 mmol, 3.7 eq) were added, and the reaction mixture was stirred for 16 h at rt. To the product mixture, water (40 mL) was added and the volume was reduced under reduced pressure to 40 mL. The solid product was filtered, washed with water (4 × 50 mL) and hexane (4 × 50 mL), and dried in vacuo to give 10. Yield 96%. 1H NMR (400 MHz, Methanol-d4) δ 8.21 (d, J = 4.4 Hz, 6H), 8.04–7.84 (m, 2H), 7.59 (s, 1H), 5.87 (d, J = 5.7 Hz, 2H), 4.86 (q, J = 7.5 Hz, 1H), 2.21 (s, 9H). HRMS Calculated [M + H]: 338.1710; Found [M + H]; 338.1361.
11 (benzyl (S)-(1,3-diamino-1-oxopropan-2-yl)carbamate). Compound 10 (1.00 g, 2.96 mmol) was solubilized in 10 ml of dichloromethane, then 10 ml of trifluoroacetic acid were added and the reaction was stirred for 1 h. The volume of the mixture was reduced under reduced pressure to 5 mL then cold diethyl ether (50 mL) was added. The suspension was centrifuged, and the precipitate (11) was further washed with diethyl ether (2 × 50 mL). Molecule 11 was then dried under vacuum and used without further purification. Yield 99% 1H NMR (400 MHz, Methanol-d4) δ 7.02 (bs, 1H), 8.35–8.21 (m, 6H), 3.86 (s, 2H), 2.21 (dd, J = 13.1, 5.3 Hz, 1H), 1.98 (dd, J = 13.1, 8.0 Hz, 1H).
13 (benzyl (S)-(1-amino-3-((5-(dimethylamino)naphthalene)-1-sulfonamido)-1-oxopropan-2-yl)carbamate). To a stirred solution of molecule 11 (1.0 g, 4.20 mmol, 1 eq) and triethylamine (0.58 mL, 4.20 mmol, 1 eq) in dry dichloromethane (20 mL) at room temperature, a solution of dansyl chloride (12) (1,36 g, 5.04 mmol 1.2 eq) in dry dichloromethane (10 mL) was added during 5 min and stirring of the reaction mixture was continued for 24 h. The reaction mixture was washed with water, the solvent was removed under vacuum and the residue was column chromatographed using a gradient of DCM/MeOH from 95:5 to 80:20 as eluents. Yield 70%. 1H NMR (400 MHz, Chloroform-d/Methanol-d4) δ 8.50–8.53 (m, 1H), 8.35–8.21 (m, 1H), 8.16 (d, J = 7.5 Hz, 1H), 7.57–7.41 (m, 2H), 7.29 (d, J = 4.8 Hz, 5H), 7.17 (d, J = 7.5 Hz, 1H), 5.00 (d, J = 3.3 Hz, 2H), 4.18 (t, J = 5.8 Hz, 1H), 3.18 (dt, J = 13.1, 6.3 Hz, 2H), 2.84 (s, 6H). 13C NMR (101 MHz, Chloroform-d/Methanol-d4) δ 173.90, 157.38, 152.52, 136.86, 135.39, 131.07, 130.58, 130.15, 129.84, 129.01, 128.88, 128.66, 128.42, 123.78, 119.51, 115.94, 67.59, 55.19, 45.69, 44.68. HRMS Calculated [M + H]: 471.1697; Found [M + H]; 471.1689.
14 ((S)-2-amino-3-((5-(dimethylamino)naphthalene)-1-sulfonamido)propanamide). To a solution of compound 13 (1.0 g, 2.12 mmol) in THF/MeOH 1:1 (20 mL), was added 10% palladium on carbon (150 mg), and the mixture was stirred under an atmosphere of H2 for 12 h. The suspension was filtered through a pad of celite and the filtrate was concentrated under reduced pressure giving a yellowish oil (14). Molecule 14 was then dried under vacuum and used without further purification. Yield 99%. 1H NMR (400 MHz, Chloroform-d/Methanol-d4) δ 8.56 (d, J = 8.5 Hz, 1H), 8.34 (d, J = 8.5 Hz, 1H), 8.30–8.17 (m, 1H), 7.65–7.46 (m, 2H), 7.22 (d, J = 7.5 Hz, 1H), 4.13 (d, J = 8.5 Hz, 1H), 3.34 (s, 2H), 2.90 (s, 6H). 13C NMR (101 MHz, Chloroform-d/Methanol-d4) δ 171.07, 152.26, 134.49, 131.05, 130.25, 129.78, 129.68, 128.92, 123.58, 119.18, 115.83, 53.97, 45.59, 44.49. HRMS Calculated [M + H]: 337.1329; Found [M + H]; 337.1325.
15 (tert-butyl ((S)-1-(((S)-1-amino-3-((5-(dimethylamino)naphthalene)-1-sulfonamido)-1-oxopropan-2-yl)amino)-5-(3-(benzyloxy)-2-methyl-4-oxopyridin-1(4H)-yl)-1-oxopentan-2-yl)carbamate). To a stirred solution of compound 8 (1.0 g, 2.32 mmol, 1.0 eq) and hydroxybenzotriazole (HOBt) (376 mg, 2.78 mmol, 1.2 eq) in DMF (50 ml) was added 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (432 mg, 2.78 mmol, 1.2 eq) at 0 °C. The mixture was stirred at this temperature for 5 min. Molecule 14 (780 mg, 2.32 mmol, 1.0 eq) and triethylamine (0.308 mL, 2.32 mmol, 1.0 eq) were then added to the solution. The resulting mixture was stirred at room temperature for 24 h. After evaporation of the solvent, the crude mixture was washed with H2O and extracted with DCM. The organic phase was dried over sodium sulfate and dried. The residue was column chromatographed on silica gel using a gradient of DCM/MeOH from 96:4 to 90:10 as eluents. Yield 90%. 1H NMR (400 MHz, Methanol-d4) δ 8.52 (dt, J = 8.5, 1.1 Hz, 1H), 8.31 (d, J = 8.5 Hz, 1H), 8.14 (dd, J = 7.4, 1.1 Hz, 1H), 7.65 (d, J = 7.4 Hz, 1H), 7.54 (ddd, J = 8.5, 7.4, 6.0 Hz, 2H), 7.35 (dd, J = 5.1, 2.0 Hz, 2H), 7.29 (dd, J = 5.1, 2.0 Hz, 2H), 7.24–7.20 (m, 1H), 6.46 (d, J = 7.4 Hz, 1H), 5.03 (s, 2H), 4.41 (t, J = 5.5 Hz, 1H), 4.08–3.77 (m, 3H), 3.21 (dd, J = 5.6, 3.4 Hz, 2H), 2.83 (s, 6H), 2.16 (s, 3H), 1.90–1.54 (m, 4H), 1.46 (s, 9H). 13C NMR (101 MHz, Methanol-d4) δ 173.21, 172.66, 157.04, 151.86, 145.66, 143.72, 139.77, 137.06, 135.06, 130.05, 129.84, 129.43, 128.81, 128.70, 128.03, 127.98, 127.93, 122.97, 118.91, 116.06, 115.15, 79.92, 73.15, 54.97, 53.35, 53.14, 44.43, 43.34, 27.85, 27.44, 26.51, 11.52. HRMS Calculated [M + H]: 749.3327; Found [M + H]; 749.3318.
16 ((S)-2-amino-N-((S)-1-amino-3-((5-(dimethylamino)naphthalene)-1-sulfonamido)-1-oxopropan-2-yl)-5-(3-(benzyloxy)-2-methyl-4-oxopyridin-1(4H)-yl)pentanamide). Compound 15 (1.00 g, 1.33 mmol) was solubilized in 10 mL of dichloromethane, then 10 mL of trifluoroacetic acid were added and the reaction was stirred for 1 h. The volume of the mixture was reduced under reduced pressure to 5 mL then cold diethyl ether (50 mL) was added. The suspension was centrifuged, and the precipitate (16) was further washed with diethyl ether (2 × 50 mL). Molecule 16 was then dried under vacuum and used without further purification. Yield 99%. 1H NMR (400 MHz, Methanol-d4) δ 8.67 (d, J = 8.4 Hz, 1H), 8.55 (d, J = 8.4 Hz, 1H), 8.32 (dd, J = 16.0, 7.2 Hz, 2H), 7.88–7.62 (m, 3H), 7.38 (dd, J = 22.4, 5.2 Hz, 5H), 7.23 (d, J = 7.2 Hz, 1H), 4.48 (s, 1H), 4.37 (d, J = 7.2 Hz, 2H), 4.05 (s, 1H), 3.24 (d, J = 9.5 Hz, 8H), 2.52 (s, 3H), 2.00 (q, J = 6.1, 5.6 Hz, 4H). 13C NMR (101 MHz, Methanol-d4) δ 173.26, 169.66, 165.86, 151.15, 145.04, 142.99, 137.29, 136.84, 130.68, 130.62, 129.99, 129.83, 129.81, 129.66, 129.59, 129.22, 125.63, 123.14, 118.04, 117.42, 114.60, 114.23, 76.20, 56.87, 55.00, 53.72, 46.45, 46.44, 44.67, 28.89, 26.34, 13.64. HRMS Calculated [M + H]: 649.2803; Found [M + H]; 649.2801.
17 (tert-butyl ((S)-1-(((R)-1-(((S)-1-(((S)-1-amino-3-((5-(dimethylamino)naphthalene)-1-sulfonamido)-1-oxopropan-2-yl)amino)-5-(3-(benzyloxy)-2-methyl-4-oxopyridin-1(4H)-yl)-1-oxopentan-2-yl)amino)-1-oxo-5-(3-((2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-5-yl)sulfonyl)guanidino)pentan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamate). To a stirred solution of compound 5 (1.0 g, 1.48 mmol, 1.0 eq) and hydroxybenzotriazole (HOBt) (240 mg, 1.78 mmol, 1.2 eq) in DMF (50 mL) was added 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (276 mg, 1.78 mmol, 1.2 eq) at 0 °C. The mixture was stirred at this temperature for 5 min. Molecule 16 (962 mg, 1.48 mmol, 1.0 eq) and triethylamine (0.206 mL, 1.48 mmol, 1.0 eq) were then added to the solution. The resulting mixture was stirred at room temperature for 24 h. After evaporation of the solvent, the crude mixture was washed with H2O and extracted with DCM. The organic phase was dried over sodium sulfate and dried. The residue was column chromatographed on silica gel using EtOH/NH4OH/DCM/Pet. Et. 13.7:1.7:71.9:12.7 as eluent. Yield 90%. 1H NMR (400 MHz, Methanol-d4) δ 8.53 (dd, J = 8.7, 5.3 Hz, 1H), 8.31 (d, J = 8.7 Hz, 1H), 8.20–8.09 (m, 1H), 7.68 (t, J = 7.0 Hz, 1H), 7.58–7.46 (m, 2H), 7.38–7.33 (m, 2H), 7.29 (tt, J = 4.8, 2.7 Hz, 3H), 7.25–7.08 (m, 6H), 6.50–6.41 (m, 1H), 5.06–4.96 (m, 2H), 4.42 (dt, J = 10.1, 3.7 Hz, 2H), 4.31 (td, J = 10.1, 9.6, 6.9 Hz, 2H), 3.94 (t, J = 6.9 Hz, 2H), 3.30–3.04 (m, 4H), 2.84 (s, 10H), 2.49–2.58 (m, 5H), 2.19 (d, J = 2.1 Hz, 3H), 2.12–1.41 (m, 11H), 1.45–1.17 (m, 16H). 13C NMR (101 MHz, Methanol-d4) δ 173.32, 173.16, 172.52, 172.33, 172.12, 171.97, 158.46, 156.79, 151.86, 145.74, 143.66, 139.83, 138.04, 137.08, 135.02, 132.94, 132.17, 130.05, 129.82, 129.42, 129.01, 128.76, 128.73, 128.13, 128.01, 127.89, 126.30, 124.72, 124.64, 122.99, 118.90, 117.05, 116.11, 115.15, 86.30, 86.26, 79.40, 73.13, 55.91, 53.66, 53.29, 53.14, 52.92, 44.42, 44.40, 43.53, 42.56, 27.67, 27.33, 26.57, 18.42, 18.36, 17.18, 11.60, 11.56, 11.52, 11.22, 11.19. HRMS Calculated [M + H]: 1304.5842; Found [M + H]; 1304.5851.
1 (BP19) ((S)-N-((S)-1-amino-3-((5-(dimethylamino)naphthalene)-1-sulfonamido)-1-oxopropan-2-yl)-2-((R)-2-((S)-2-amino-3-phenylpropanamido)-5-guanidinopentanamido)-5-(3-hydroxy-2-methyl-4-oxopyridin-1(4H)-yl)pentanamide). Compound 17 (1.00 g, 0.76 mmol) was solubilized in 20 ml of trifluoroacetic acid and the reaction was stirred for 24 h. The volume of the mixture was reduced under reduced pressure to 5 mL then cold diethyl ether (50 mL) was added. The suspension was centrifuged, and the precipitate (1) was further washed with diethyl ether (2 × 50 mL). Molecule 1 was then dried under vacuum and used without further purification for biophysical studies. Samples used in biological assays were further purified by preparative HPLC. Yield 99%. 1H NMR (400 MHz, Methanol-d4/D2O/CD3COCD3) δ 8.54 (d, J = 8.5 Hz, 1H), 8.35 (q, J = 8.5, 7.8 Hz, 1H), 8.17 (d, J = 7.8 Hz, 1H), 8.04 (s, 1H), 7.64 (p, J = 8.5, 7.8 Hz, 2H), 7.43–7.16 (m, 6H), 7.03 (s, 1H), 4.55–4.14 (m, 6H), 3.30–2.82 (m, 12H), 2.58 (s, 3H), 1.93 (s, 6H), 1.66 (s, 2H). 13C NMR (101 MHz, Chloroform-d/Methanol-d4) δ 172.89, 172.60, 169.26, 168.84, 161.91, 156.93, 134.78, 133.85, 129.73, 129.59, 129.41, 129.34, 129.00, 128.94, 128.37, 128.32, 127.66, 127.61, 123.95, 120.19, 116.20, 55.51, 54.06, 53.40, 45.22, 43.42, 40.60, 36.89, 30.14, 28.51, 27.68, 26.51, 24.49, 11.89. HRMS Calculated [M + H]: 862.4029; Found [M + H]; 862.4016.
References
Abbate V, Reelfs O, Hider RC, Pourzand C (2015) Design of novel fluorescent mitochondria-targeted peptides with iron-selective sensing activity. Biochem J 469:357–366. https://doi.org/10.1042/BJ20150149
Abbate V, Reelfs O, Kong X, Pourzand C, Hider RC (2016) Dual selective iron chelating probes with a potential to monitor mitochondrial labile iron pools. Chem Commun 52:784–787. https://doi.org/10.1039/C5CC06170A
Alta RYP, Vitorino HA, Goswami D, Machini MT, Espósito BP (2017a) Triphenylphosphonium-desferrioxamine as a candidate mitochondrial iron chelator. Biometals 30:709–718. https://doi.org/10.1007/s10534-017-0039-5
Alta RYP, Vitorino HA, Goswami D, Liria CW, Wisnovsky SP, Kelley SO, Machini MT, Espósito BP (2017b) Mitochondria-penetrating peptides conjugated to desferrioxamine as chelators for mitochondrial labile iron. PLoS ONE 12(2):e0171729. https://doi.org/10.1371/journal.pone.0171729
Altamura S, Muckenthaler MU (2009) Iron toxicity in diseases of aging: Alzheimer’s disease, Parkinson’s disease and atherosclerosis. J Alzheimer’s Dis 16:879–895. https://doi.org/10.3233/JAD-2009-1010
Aroun A, Zhong JL, Tyrrell RM, Pourzand C (2012) Iron, oxidative stress and the example of solar ultraviolet A radiation. Photochem Photobiol Sci 11:118–134. https://doi.org/10.1039/C1PP05204G
Boddaert N, Le Quan Sang KH, Rötig A, Leroy-Willig A, Gallet S, Brunelle F, Sidi D, Thalabard JC, Munnich A, Cabantchik ZI (2007) Selective iron chelation in Friedreich ataxia: biologic and clinical implications. Clinical Trial 110:401–408. https://doi.org/10.1182/blood-2006-12-065433
Chiang S, Huang MLH, Park KC, Richardson DR (2020) Antioxidant defense mechanisms and its dysfunctional regulation in the mitochondrial disease, Friedreich’s ataxia. Free Radic Biol Med 159:177–188. https://doi.org/10.1016/j.freeradbiomed.2020.07.019
Cilibrizzi A, Abbate V, Chen Y-L, Ma Y, Zhou T, Hider RC (2018) Hydroxypyridinone journey into metal chelation. Chem Rev 118:7657–7701. https://doi.org/10.1021/acs.chemrev.8b00254
Cloonan SM, Glass K, Laucho-Contreras ME, Bhashyam AR, Cervo M et al (2016) Mitochondrial iron chelation ameliorates cigarette smoke-induced bronchitis and emphysema in mice. Nat Med 22(2):163–174. https://doi.org/10.1038/nm.4021
Connor JR, Snyder BS, Beard JL, Fine RE, Mufson EJ (1992) Regional distribution of iron and iron-regulatory proteins in the brain of in aging and Alzheimer’s disease. J Neurosci Res 31:327–335. https://doi.org/10.1002/jnr.490310214
Deus CM, Pereira SP, Cunha-Oliveira T, Teixeira J, Simões RF, Cagide F, Benfeito S, Borges F, Raimundo N, Oliveira PJ (2021) A mitochondria-targeted caffeic acid derivative reverts cellular and mitochondrial defects in human skin fibroblasts from male sporadic Parkinson’s disease patients. Redox Biol 45:102037. https://doi.org/10.1016/j.redox.2021.102037
Devos D, Moreau C, Devedjian JC et al (2014) Targeting chelatable iron as a therapeutic modality in Parkinson’s disease. Antioxid Redox Signal 21:195–210. https://doi.org/10.1089/ars.2013.5593
Dexter DT, Wells FR, Agid F, Agid Y, Lees AJ, Jenner P, Marsden CD (1987) Increased nigral iron content in postmortem parkinsonian brain. Lancet 2:1219–1220. https://doi.org/10.1016/s0140-6736(87)91361-4
Dissemond J, Schneider LA, Brenneisen P, Briviba K, Wenk J, Wlaschek M, Scharffetter-Kochanek K (2003) Protective and determining factors for the overall lipid peroxidation in ultraviolet al-irradiated fibroblasts: in vitro and in vivo investigations. Br J Dermatol 149:341–349. https://doi.org/10.1046/j.1365-2133.2003.05457.x
Futaki S (2006) Oligoarginine vectors for intracellular delivery: design and cellular-uptake mechanisms. Biopolymers 84:241–249. https://doi.org/10.1002/bip.20421
Gao J, Zhou Q, Wu D, Chen L (2021) Mitochondrial iron metabolism and its role in diseases. Clin Chim Acta 513:6–12. https://doi.org/10.1016/j.cca.2020.12.005
Hider RC, Kong X (2010) Chemistry and biology of siderophores. Nat Prod Rep 27:637–657. https://doi.org/10.1039/B906679A
Hirayama T, Okuda K, Nagasawa H (2013) A highly selective turn-on fluorescent probe for iron(II) to visualize labile iron in living cells. Chem Sci 4:1250–1256. https://doi.org/10.1039/C2SC21649C
Hirayama T, Tsuboi H, Niwa M, Miki A, Kadota S, Ikeshita Y, Okuda K, Nagasawa H (2017) A universal fluorogenic switch for Fe(II) ion based on N-oxide chemistry permits the visualization of intracellular redox equilibrium shift towards labile iron in hypoxic tumor cells. Chem Sci 8:4858–4866. https://doi.org/10.1039/C6SC05457A
Hirayama T, Miki A, Nagasawa H (2019) Organelle-specific analysis of labile Fe(II) during ferroptosis by using a cocktail of various colour organelle-targeted fluorescent probes. Metallomics 11:111–117. https://doi.org/10.1039/C8MT00212F
Horton KL, Stewart KM, Fonseca SB, Guo Q, Kelley SO (2008) Mitochondria-penetrating peptides. Chem Biol 15:375–382. https://doi.org/10.1016/j.chembiol.2008.03.015
Ke Y, Qian ZM (2003) Iron misregulation in the brain: a primary cause of neurodegenerative disorders. Lancet Neurol 2:246–253. https://doi.org/10.1016/S1474-4422(03)00353-3
Krig D, Carroll MT, Cooper JM, Marsden CD, Schapira AJ (1992) Platelet mitochondrial function in-Parkinson’s disease. The Royal Kings and Queens Parkinson Disease Research Group. Ann Neurol 32:782–788. https://doi.org/10.1002/ana.410320612
Lim CK, Kalinowski DS, Richardson DR (2008) Protection against hydrogen peroxide-mediated cytotoxicity in Friedreich’s ataxia fibroblassts using novel iron chelators of the 2-pyridylcarboxaldehyde isonicotionoyl hydrazone class. Mol Pharmacol 74:225–235. https://doi.org/10.1124/mol.108.046847
Lin MT, Beal MF (2006) Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature 443:787–795. https://doi.org/10.1038/nature05292
Llorens JV, Soriano S, Calap-Quintana P, Gonzalez-Cabo P, Moltó MD (2019) The role of iron in Friedreich’s ataxia: insights from studies in human tissues and cellular and animal models. Front Neurosci 13:75. https://doi.org/10.3389/fnins.201900075
Martin V, Egelund PHG, Johansson H, Le Quement ST, Wojcik F, Pedersen DS (2020) Greening the synthesis of peptide therapeutics: an industrial perspective. RSC Adv 10:42457–42492. https://doi.org/10.1039/D0RA07204D
Mena NP, Urrutia PJ, Lourido F, Carrasco CM, Núñez MT (2015) Mitochondrial iron homeostasis and its dysfunctions in neurodegenerative disorders. Mitochondrion 21:92–105. https://doi.org/10.1016/j.mito.2015.02.001
Murphy MP (2008) Targeting lipophilic cations to mitochondria. Biochem Biophys Acta 1777:1028–1031. https://doi.org/10.1016/j.bbabio.2008.03.029
Nakase I, Okumura S, Katayama S, Hirose H, Pujals S, Yamaguchi H, Arakawa S, Shimizu S, Futaki S (2012) Transformation of an antimicrobial peptide into a plasma membrane-permeable, mitochondria-targeted peptide via the substitution of lysine with arginine. Chem Commun 48:11097–11099. https://doi.org/10.1039/c2cc35872g
Paul VD, Lill R (2015) Biogenesis of cytosolic and nuclear iron-sulfur proteins and their role in genome stability. Biochim Biophys Acta 1853:1528–1539. https://doi.org/10.1016/j.bbamcr.2014.12.018
Ponka P (1999) Cell biology of heme. Am J Med Sci 318:241–256. https://doi.org/10.1016/S0002-9629(15)40628-7
Pourzand C, Reelfs O, Allen C, Hussain K, Muhammad J, Tomy H, Chen Y-L, Cilibrizzi A, Abbate V, Hider RC (2019). The cytoprotective potential of novel mitochondria-targeted iron chelators against hydrogen peroxide-mediated toxicity in Friedreich’s ataxia fibroblasts. In: Abstract in EMBL Conference—8th Congress of the International BioIron Society, May 2019, Heidelberg, Germany, p 374
Reelfs O, Abbate V, Hider RC, Pourzand C (2016) A powerful mitochondria-targeted iron chelator affords high photoprotection against solar ultraviolet A radiation. J Invest Dermatol 136:1692–1700. https://doi.org/10.1016/j.jid.2016.03.041
Reelfs O, Abbate V, Cilibrizzi A, Pook MA, Hider RC, Pourzand C (2019a) The role of mitochondrial labile iron in Friedreich’s ataxia skin fibroblasts sensitivity to ultraviolet A. Metallomics 11:656–665. https://doi.org/10.1039/c8mt00257f
Reelfs O, Chen Y-L, Abbate V, Hider RC, Pourzand C (2019b) Mitochondrial-targeted iron chelators for the therapy of Parkinson’s disease. In: Abstract in EMBL Conference—8th Congress of the International BioIron Society, May 2019b, Heidelberg, Germany, p373.
Rouault TA (2016) Mitochondrial iron overload: causes and consequences. Curr Opin Genet Dev 38:31–37. https://doi.org/10.1016/j.gde.2016.02.004
Sandoval-Acuña C, Torrealba N, Tomkova V, Jadhav SB, Blazkova K, Merta L, Lettlova S, Adamcová MK, Rosel D, Brábek J, Neuzil J, Stursa J, Werner L, Truksa J (2021) Targeting mitochondrial iron metabolism suppresses tumor growth and metastasis by inducing mitochondrial dysfunction and mitophagy. Cancer Res 81:2289–2303. https://doi.org/10.1158/0008-5472.CAN-20-1628
Schiller PW, Nguyen TM, Berezowska I, Dupuis S, Weltrowska G, Chung NN, Lemieux C (2000) Synthesis and in vitro opioid activity profiles of DALDA analogues. Eur J Med Chem 35:895–901. https://doi.org/10.1016/s0223-5234(00)01171-5
Smith RAJ, Porteous CM, Gane AM, Murphy MP (2003) Delivery of bioactive molecules to mitochondria in vivo. Proc Natl Acad Sci USA 100:5407–5412. https://doi.org/10.1073/pnas.0931245100
Talukdar P, Bollot G, Mareda J, Sakai N, Matile S (2005) Ligand-gated synthetic ion channels. Chemistry 11:3525–3532. https://doi.org/10.1002/chem.200500516
Urrutia PJ, Mena NP, Nunez MT (2014) The interplay between iron accumulation, mitochondrial dysfunction and inflammation during the execution step of neurodegenerative disorders. Front Pharmacol 5:38. https://doi.org/10.3389/fphar.2014.00038 (eCollection 2014)
Urrutia PJ, Bórquez DA, Núñez MT (2021) Inflaming the brain with iron. Antioxidants (basel) 10:61. https://doi.org/10.3390/antiox10010061
Victor VM, Rocha M (2007) Targeting antioxidants to mitochondria: a potential new therapeutic strategy for cardiovascular disease. Curr Pharm Des 13:845–863. https://doi.org/10.2174/138161207780363077
Vile GF, Tyrrell RM (1995) UVA radiation-induced oxidative damage to lipids and proteins in vitro and in human skin fibroblasts is dependent on iron and singlet oxygen. Free Radic Biol Med 18:721–730. https://doi.org/10.1016/0891-5849(94)00192-m
Wong A, Yang J, Cavadini P, Gellera C, Lonnerdal B, Taroni F, Cortopassi G (1999) The Friedreich’s ataxia mutation confers cellular sensitivity to oxidant stress which is rescued by chelators of iron and calcium and inhibitors of apoptosis. Hum Mol Genet 8:425–430. https://doi.org/10.1093/hmg/8.3.425
Zhao GM, Qian X, Schiller PW, Szeto HH (2003) Comparison of [Dmt1]DALDA and DAMGO in binding and G protein activation at μ, δ, and κ opioid receptors. J Pharmacol Exp Ther 307:947–954. https://doi.org/10.1124/jpet.103.054775
Zhong JL, Yiakouvaki A, Holley P, Tyrrell RM, Pourzand C (2004) Susceptibility of skin cells to UVA-induced necrotic cell death reflects the intracellular level of labile iron. J Invest Dermatol 123:771–780. https://doi.org/10.1111/j.0022-202X.2004.23419.x
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We wish to thank the BBSRC (Grant No. BB/J005223/1) and Parkinson’s UK (Grant No. K-1603) for the generous support of this work.
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Dedication: This paper is dedicated to our colleague Professor Dr. Günther Winkelman on the occasion of his 80th birthday and his retirement as long-term editor of BioMetals. One of us (RH) first met Günther in 1977 and has been in continuous contact since that date on many matters relating to siderophores.
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Cilibrizzi, A., Pourzand, C., Abbate, V. et al. The synthesis and properties of mitochondrial targeted iron chelators. Biometals 36, 321–337 (2023). https://doi.org/10.1007/s10534-022-00383-8
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DOI: https://doi.org/10.1007/s10534-022-00383-8