Next Article in Journal
Statistical Optimization of Medium for Pullulan Production by Aureobasidium pullulans NCPS2016 Using Fructose and Soybean Meal Hydrolysates
Next Article in Special Issue
Ferrocenes as Building Blocks in Molecular Rectifiers and Diodes
Previous Article in Journal
Aromaticity as a Guiding Concept for Spectroscopic Features and Nonlinear Optical Properties of Porphyrinoids
Previous Article in Special Issue
Enantiomerically Enriched 1,2-P,N-Bidentate Ferrocenyl Ligands for 1,3-Dipolar Cycloaddition and Transfer Hydrogenation Reactions
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
Molecules
Volume 23
Issue 6
10.3390/molecules23061335
molecules-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Trapping para-Quinone Methide Intermediates with Ferrocene: Synthesis and Preliminary Biological Evaluation of New Phenol-Ferrocene Conjugates

by
Silvia González-Pelayo
1,
Enol López
1,
Javier Borge
2,
Noemí De-los-Santos-Álvarez
3 and
Luis A. López
1,*
1
Departamento de Química Orgánica e Inorgánica and Instituto Universitario de Química Organometálica “Enrique Moles”, Universidad de Oviedo, Julián Clavería 8, 33006 Oviedo, Spain
2
Laboratorio de Compuestos Organometálicos y Catálisis (Unidad Asociada al CSIC), Centro de Innovación en Química Avanzada (ORFEO-CINQA) and Departamento de Química Física y Analítica, Universidad de Oviedo, Julián Clavería 8, 33006 Oviedo, Spain
3
Departamento de Química Física y Analítica, Universidad de Oviedo, Julián Clavería 8, 33006 Oviedo, Spain
*
Author to whom correspondence should be addressed.
Molecules 2018, 23(6), 1335; https://doi.org/10.3390/molecules23061335
Submission received: 9 May 2018 / Revised: 29 May 2018 / Accepted: 29 May 2018 / Published: 1 June 2018
(This article belongs to the Special Issue Ferrocene and Ferrocene-Containing Compounds)
Browse Figures
Versions Notes

Abstract

:
The reaction of para-hydroxybenzyl alcohols with ferrocene in the presence of a catalytic amount of InCl3 provided ferrocenyl phenol derivatives, an interesting class of organometallic compounds with potential applications in medicinal chemistry. This transformation exhibited a reasonable substrate scope delivering the desired products in synthetically useful yields. Evidence of involvement of a para-quinone methide intermediate in this coupling process was also provided. Preliminary biological evaluation demonstrated that some of the ferrocene derivatives available by this methodology exhibit significant cytotoxicity against several cancer cell lines with IC50 values within the range of 1.07–4.89 μM.

1. Introduction

Since the discovery of ferrocene in the 1950s [1,2], the interest in this organometallic compound has not declined. In fact, its chemistry remains one of the most active areas of research. Very likely, this enduring interest resides in the fact that many functionalized ferrocene derivatives display a wide number of applications in a diverse range of fields [3,4,5,6,7,8,9]. For example, recent investigations have demonstrated the potential of some ferrocene derivatives in medicinal chemistry [10,11]. Particularly, some ferrocene-containing phenols have proved to be of great interest in cancer therapeutics because of their antitumoral activity [12,13,14,15,16]. Among them, a family of ferrocene analogues of hydroxytamoxifen, the so-called ferrocifens (Figure 1a), have been the subject of in-depth investigations showing exceptional cytotoxic activities against some types of breast cancer [17,18,19]. The mode of action of these organometallic drug candidates has been elucidated by electrochemical and chemical oxidation methods. According to these studies, ferrocenyl quinone methides have been suggested to play a key role in the antiproliferative activity [20,21,22,23,24]. The antitumoral activity of some unconjugated bisphenol derivatives of ferrocene (Figure 1b) has also been evaluated [25,26].
In connection with our studies on C–H bond functionalization of ferrocene based on the trapping of highly electrophilic species [27,28,29], we have recently described the trapping of ortho-quinone methide intermediates with ferrocene [30]. Interestingly, some of the ferrocene-containing monophenol derivatives available by this methodology (Figure 1c) display remarkable cytotoxic activity against various cancer cell lines.
In order to elucidate the structural requirements for cytotoxicity and, eventually, to identify more bioactive derivatives, we decided to develop a synthetic methodology for the synthesis of the isomeric para-substituted ferrocenylphenol analogues (Figure 1d). Herein, we report the results of this study; specifically, we describe the generation of para-quinone methide intermediates and their trapping with ferrocene. Preliminary biological evaluation of some of the functionalized ferrocene derivatives prepared in this study is also disclosed.

2. Results and Discussion

The present study was carried out using easily available p-hydroxybenzyl alcohols 1a–l outlined in Figure 2.
For our initial study, p-hydroxybenzyl alcohol 1a was chosen as the model substrate (Scheme 1). On the basis of our previous investigations in the ortho series, InCl3 in dichloroethane (DCE) was selected as the catalytic system. Pleasingly, we found that heating a mixture of 1a (1 equiv.), ferrocene (2, 3 equiv.), and InCl3 (10 mol%) in DCE at 60 °C led to complete disappearance of the starting p-hydroxybenzyl alcohol after 2 h (checked by thin layer chromatography, TLC). Chromatographic purification (SiO2, 5:1 hexane ethyl acetate) provided the desired functionalized ferrocene derivative 3a in a remarkable 75% yield. Interestingly, under otherwise similar conditions, benzydryl alcohol 1a’ was found to be a fruitless reaction partner, thus demonstrating the key role of the phenolic OH group in the reaction course [31,32].
Ferrocenyl phenol 3a was characterized by Nuclear Magnetic Resonance (NMR) spectroscopy. Moreover, crystals of compound 3a were obtained from diffusion of pentane into a dichloromethane solution at −20 °C and its molecular structure in the solid state has been determined by single-crystal X-ray diffraction (Figure 3 and Appendix A) [33,34]. The electrochemical behavior of compound 3a was investigated by cyclic voltammetry (Appendix B).
With suitable reaction conditions in hand (10 mol% of InCl3, DCE as solvent, 60 °C), a variety of p-hydroxybenzyl alcohols were then evaluated for their suitability for this C–H bond functionalization process (Table 1). First, some p-hydroxybenzyl alcohols substituted with various aryl groups were investigated (entries 2–5). As shown, all three isomeric 4-[hydroxy(tolyl)methyl]phenol derivatives 1b–d (R = tolyl) served as suitable reaction partners for this process furnishing the desired functionalized ferrocene derivatives 3b–d in acceptable isolated yields (51–82%, entries 2–4). Similarly, para-methoxy substituted substrate 1e (R = p-MeOC6H4) delivered the corresponding product 3e in moderate isolated yield (48%, entry 5).
Next, substrates 1f–j with alkyl groups in the benzylic position were tested (entries 6–10). As shown, primary, secondary and tertiary alkyl groups were well tolerated delivering functionalized ferrocene derivatives 3f–j in moderate yields (42–62%).
This transformation was compatible with unsaturated functional groups in the benzylic position as demonstrated by the synthesis of ferrocene derivative 3k in moderate yield when substrate 1k (R = allyl) was subjected to the standard reaction conditions. Finally, we found that the parent p-hydroxybenzyl alcohol 1l (R = H) is also a viable substrate affording the desired product 3l in 64% yield (entry 12).
Next, to provide further evidence for the involvement of p-quinone methide intermediates in the present coupling, we performed an experiment with a stable p-quinone methide. Thus, 4-benzylidene-2,6-di-tert-butylcyclohexa-2,5-dienone (4) and ferrocene (2) were subjected to the standard reaction conditions (10 mol% of InCl3, DCE, 60 °C). However, a very low conversion was observed after 24 h very likely due to steric hindrance by the tert-butyl groups. Gratifyingly, performing the reaction in toluene at 100 °C enabled the preparation of ferrocene derivative 5 in 80% yield (Scheme 2). Notably, in the absence of InCl3, no reaction occurred at all.
Based on these control experiments and on previous related literature precedents, a mechanistic proposal for the reaction of hydroxybenzyl alcohols 1 and ferrocene (2) is outlined in Scheme 3. In the present process, the Lewis acid is proposed to play a dual role. Firstly, it would promote the generation of the key quinone methide intermediate through dehydration of hydroxybenzyl alcohol 1. Subsequent activation of the quinone methide by Lewis acid complexation would provide intermediate I. This intermediate, with a high electrophilic character at the exocyclic C=C bond, may be involved in a Friedel-Crafts type electrophilic aromatic substitution [35]. Indeed, 1,6-addition of ferrocene would provide the σ-complex intermediate II, which would evolve to the final product with regeneration of the catalyst [36].
Some of the functionalized ferrocene derivatives prepared were evaluated for their cytotoxicity against several cancer cell lines (Table 2). In this preliminary study, ferrocene derivatives 3a and 3g were identified as the most active ones [37]. For example, 3a displayed significant toxicity against A2780 ovarian cancer cell line (IC50 of 1.07 μM). Compared with the value previously reported for the ortho-isomer (IC50 of 2.68 μM), ferrocene derivative 3a has superior characteristics. Ferrocene 3g also displayed toxicity against this cell line (IC50 of 2.23 μM), although somewhat lower than that found for the ortho-analogue (IC50 of 1.86 μM). We have also studied the cytotoxicity profile of ferrocene derivatives 3a and 3g against A549 lung cancer cells. Both derivatives exhibited moderate cytotoxicity with IC50 values of 3.55 and 4.89 μM, respectively. These values are comparable to that previously measured for the ortho-isomers (IC50 of 2.77 and 5.96 μM, respectively).

3. Materials and Methods

3.1. General

NMR spectra were recorded at room temperature in CDCl3 on a Bruker DPX-300 or Bruker AVANCE-300 MHz instruments (Bruker, Billerica, MA, USA). Chemical shifts are given in ppm relative to TMS (1H, 0.0 ppm) or CDCl3 (13C, 77.0 ppm). High-resolution mass spectra were determined on a VG Autospec M mass spectrometer (Waters Corporation, Milford, MA, USA). Cyclic voltammetric studies were performed using a µ-AutoLab type II equipped with GPES 4.9 software (EcoChemie, Utrecht, The Netherlands). All measurements were carried out using a conventional three electrode system in phosphate saline buffer (pH 7.4). A modified carbon paste acted as the working electrode and a Pt wire as a counter electrode. All potentials were referred to a Ag|AgCl|KCl(sat) reference electrode.
Experiments were carried out under nitrogen using standard Schlenck techniques. 1,2-Dichloroethane was distilled from CaH2. Toluene was distilled from sodium-benzophenone ketyl prior to use. TLC was performed on aluminum-backed plates coated with silica gel 60 with F254 indicator. Flash column chromatography was carried out on silica gel (230–240 mesh). The solvents used in column chromatography, hexane and ethyl acetate, were obtained from commercial suppliers and used without further purification.
p-Hydroxybenzyl alcohols 1a–k were prepared by reaction of 4-hydroxybenzaldehyde with the corresponding Grignard reagents following a literature procedure [38]. p-Hydroxybenzyl alcohol 1l was obtained by the reaction of 4-hydroxybenzaldehyde with NaBH4 [39]. 4-Benzylidene-2,6-di-tert-butylcyclohexa-2,5-dienone (4) was prepared from 2,6-di-tert-butylphenol and benzaldehyde according to a literature procedure [40]. Ferrocene (2) was commercially available and used without further purification.

3.2. General Procedure for the Synthesis of Ferrocene Derivatives 3a–l

InCl3 (4.4 mg, 0.02 mmol, 10 mol%) was added to a solution of p-hydroxybenzyl alcohols 1 (0.2 mmol) and ferrocene 2 (111.6 mg, 0.6 mmol) in 1,2-dichloroethane (2 mL). The mixture was stirred at 60 °C for 2–14 h (disappearance of 1 checked by TLC). The solvent was removed under reduced pressure and the resulting residue was purified by flash chromatography (silica gel, mixtures of hexanes/ethyl acetate). Two fractions were collected. The first fraction was unreacted ferrocene and the second one was the corresponding functionalized ferrocene derivative 3. Crystals of compound 3a suitable for X-ray analysis were obtained from diffusion of pentane into dichloromethane at −20 °C. Copies of 1H- and 13C-NMR spectra are provided in the Supplementary Materials.
4-[(Ferrocenyl)(phenyl)methyl]phenol (3a): yellow solid; melting point (m.p.) 63–64 °C; 1H-NMR (300 MHz, CDCl3): 4.02 (s, 2H, Cp), 4.05 (s, 5H, Cp), 4.19 (s, 2H, Cp), 4.83 (s, 1H, OH), 5.13 (s, 1H, CH), 6.76 (d, J = 8.1 Hz, 2H, Ar), 7.07 (d, J = 8.1 Hz, 2H, Ar), 7.19–7.33 (m, 5H, Ar); 13C-NMR (75 MHz, CDCl3): 51.0 (CH), 67.59 (CH), 67.63 (CH), 68.7 (CH), 92.0 (C), 114.9 (CH), 126.1 (CH), 128.1 (CH), 128.7 (CH), 129.9 (CH), 137.6 (C), 145.3 (C), 153.7 (C); HRMS (EI) calculated for [C23H20FeO]+ (M+): 368.0858, found 368.0856.
4-[(Ferrocenyl)(2-methylphenyl)methyl]phenol (3b): yellow solid; m.p. 66–67 °C; 1H-NMR (300 MHz, CDCl3): 2.25 (s, 3H, CH3), 3.81 (s, 1H, Cp), 4.07 (s, 5H, Cp), 4.16–4.12 (m, 1H, Cp), 4.20–4.16 (m, 1H, Cp), 4.76 (s, 1H, Cp), 5.25 (s, 1H, CH), 6.78 (d, J = 8.6 Hz, 2H, Ar), 6.96 (s, 1H, OH), 7.09–7.14 (m, 6H, Ar); 13C-NMR (75 MHz, CDCl3): 20.3 (CH3), 47.9 (CH), 67.4 (CH), 68.5 (CH), 68.8 (CH), 69.1 (CH), 69.9 (CH), 93.4 (C), 115.1 (CH), 125.9 (CH), 126.5 (CH), 128.6 (CH), 130.5 (CH), 130.9 (CH), 135.5 (C), 136.0 (C), 144.7 (C), 154.1 (C); HRMS (EI) calculated for [C24H22FeO]+ (M+): 382.1015, found 382.1009.
4-[(Ferrocenyl)(3-methylphenyl)methyl]phenol (3c): yellow solid; m.p. 72–73 °C; 1H-NMR (300 MHz, CDCl3): 2.32 (s, 3H, CH3), 4.02–4.05 (m, 7H, Cp), 4.18 (m, 2H, Cp), 4.62 (s, 1H, OH), 5.06 (s, 1H, CH), 6.75 (d, J = 8.4 Hz, 2H, Ar), 7.00–7.17 (m, 6H, Ar); 13C-NMR (75 MHz, CDCl3): 21.5 (CH3), 50.9 (CH), 67.7 (CH), 68.9 (CH), 92.3 (C), 114.7 (CH), 120.5 (CH), 125.6 (CH), 126.8 (CH), 127.9 (CH), 129.4 (CH), 129.9 (CH), 137.5 (C), 145.2 (C), 153.7 (C); HRMS (EI) calculated for [C24H22FeO]+ (M+): 382.1015, found 382.1011.
4-[(Ferrocenyl)(4-methylphenyl)methyl]phenol (3d): yellow solid; m.p. 64–65 °C; 1H-NMR (300 MHz, CDCl3): 2.33 (s, 3H, CH3), 3.99 (m, 2H, Cp), 4.03 (s, 5H, Cp), 4.17 (m, 2H, Cp), 4.73 (s, 1H, OH), 5.08 (s, 1H, CH), 6.74 (d, J = 8.5 Hz, 2H, Ar), 7.04–7.11 (m, 6H, Ar); 13C-NMR (75 MHz, CDCl3): 21.4 (CH3), 51.0 (CH), 67.9 (CH), 69.1 (CH), 92.6 (C), 115.2 (CH), 128.9 (CH), 129.1 (CH), 130.2 (CH), 135.9 (C), 138.2 (C), 142.7 (C), 154.1 (C); HRMS (EI) calculated for [C24H22FeO]+ (M+): 382.1015, found 382.1009.
4-[(Ferrocenyl)(4-methoxyphenyl)methyl]phenol (3e): yellow oil; 1H-NMR (300 MHz, CDCl3): 3.80 (s, 3H, OMe), 3.98 (s, 2H, Cp), 4.04 (s, 5H, Cp), 4.17 (s, 2H, Cp), 4.72 (s, 1H, CH), 5.06 (s, 1H, OH), 6.79–6.73 (m, 2H, Ar), 6.80–6.84 (m, 2H, Ar), 7.03–7.10 (m, 4H, Ar); 13C-NMR (75 MHz, CDCl3): 50.5 (CH), 55.7 (CH3), 68.0 (CH), 69.2 (CH), 92.9 (C), 113.8 (CH), 115.2 (CH), 130.0 (CH), 130.2 (CH), 138.1 (C), 138.3 (C), 154.1 (C), 158.2 (C); HRMS (EI) calculated for [C24H22FeO2]+ (M+): 398.0964, found 398.0972.
4-(1-Ferrocenylethyl)phenol (3f): yellow solid; m.p. = 95–96 °C; 1H-NMR (300 MHz, CDCl3): 1.57 (d, J = 7.2 Hz, 3H, CH3), 3.90 (q, J = 7.2 Hz, 1H, CH), 4.09 (s, 1H, Cp), 4.10–4.18 (m, 8H, Cp), 4.63 (s, 1H, OH), 6.74 (d, J = 8.6 Hz, 2H, Ar), 7.05 (d, J = 8.6 Hz, 2H, Ar); 13C-NMR (75 MHz, CDCl3): 23.1 (CH3), 39.3 (CH), 66.7 (CH), 67.3 (CH), 67.9 (CH), 68.2 (CH), 69.0 (CH), 95.1 (C), 115.4 (CH), 128.6 (CH), 140.4 (C), 153.9 (C); HRMS (EI) calculated for [C18H18FeO]+ (M+): 306.0702, found 306.0701. Ferrocene 3f is a known compound; our characterization data match those previously reported in the literature [25].
4-(1-Ferrocenylpropyl)phenol (3g): yellow solid; m.p. = 87–88 °C; 1H-NMR (300 MHz, CDCl3): 0.85 (t, J = 7.4 Hz, 3H, CH3), 1.67–1.83 (m, 1H, CH2), 2.07–2.14 (m, 1H, CH2), 3.41–3.46 (dd, J = 10.7 and 4.3 Hz, 1H, CH), 4.06 (s, 1H, Cp), 4.05–4.10 (m, 7H, Cp), 4.17 (s, 1H, Cp), 4.73 (s, 1H, OH), 6.78 (d, J = 8.5 Hz, 2H, Ar), 7.06 (d, J = 8.5 Hz, 2H, Ar); 13C-NMR (75 MHz, CDCl3): 12.7 (CH3), 30.0 (CH2), 47.0 (CH), 66.8 (CH), 66.9 (CH), 67.3 (CH), 67.4 (CH), 68.6 (CH), 94.8 (C), 114.9 (CH), 129.0 (CH), 137.8 (C), 153.6 (C); HRMS (EI) calculated for [C19H20FeO]+ (M+): 320.0858, found 320.0853.
4-(1-Ferrocenylpentyl)phenol (3h): yellow oil; 1H-NMR (300 MHz, CDCl3): 0.89 (t, J = 7.0 Hz, 3H, CH3), 1.16–1.40 (m, 4H, CH2), 1.70–1.82 (m, 1H, CH2), 2.01–2.08 (m, 1H, CH2), 3.53 (dd, J = 10.8 and 4.3 Hz, 1H, CH), 3.95 (s, 1H, Cp), 4.05–4.11 (m, 7H, Cp), 4.18 (s, 1H, Cp), 4.80 (s, 1H, OH), 6.76 (d, J = 8.3 Hz, 2H, Ar), 7.06 (d, J = 8.3 Hz, 2H, Ar); 13C-NMR (75 MHz, CDCl3): 14.1 (CH3), 22.7 (CH2), 30.2 (CH2), 36.8 (CH2), 45.1 (CH), 66.8 (CH), 66.9 (CH), 67.3 (CH), 67.4 (CH), 68.6 (CH), 95.1 (C), 114.9 (CH), 128.9 (CH), 138.1 (C), 153.6 (C); HRMS (EI) calculated for [C21H24FeO]+ (M+): 348.1171, found 348.1184.
4-[1-(Ferrocenyl)(2-methyl)propyl]phenol (3i): yellow solid; m.p. = 95–96 °C; 1H-NMR (300 MHz, CDCl3): 0.74 (d, J = 6.6 Hz, 3H, CH3), 0.92 (d, J = 6.6 Hz, 3H, CH3), 1.97–2.03 (m, 1H, CH), 3.08 (d, J = 8.7 Hz, 1H, CH), 3.94 (s, 5H, Cp), 4.13–4.27 (m, 4H, Cp), 4.74 (s, 1H, OH), 6.75 (d, J = 8.3 Hz, 2H, Ar), 7.05 (d, J = 8.3 Hz, 2H, Ar); 13C-NMR (75 MHz, CDCl3): 21.7 (CH3), 22.2 (CH3), 53.1 (CH), 66.7 (CH), 67.2 (CH), 68.4 (CH), 69.3 (CH), 70.4 (CH), 94.7 (C), 114.9 (CH), 129.6 (CH), 137.8 (C), 153.5 (C); HRMS (EI) calculated for [C20H22FeO]+ (M+): 334.1015, found 334.1012.
4-[1-(Ferrocenyl)(2,2-dimethyl)propyl]phenol (3j): yellow solid; m.p. = 105–106 °C; 1H-NMR (300 MHz, CDCl3): 0.84 (s, 9H, CH3), 3.23 (s, 1H, CH), 3.73 (s, 5H, Cp), 4.07 (s, 1H, Cp), 4.14 (d, J = 6.2 Hz, 2H, Cp), 4.24 (s, 1H, Cp), 4.93 (s, 1H, OH), 6.84 (d, J = 8.4 Hz, 2H, Ar), 7.27–7.29 (m, 2H, Ar); 13C-NMR (75 MHz, CDCl3): 28.7 (CH3), 35.2 (C), 57.3 (CH), 65.7 (CH), 68.0 (CH), 68.1 (CH), 68.5 (CH), 69.2 (CH), 72.1 (CH), 90.4 (C), 114.2 (CH), 132.0 (CH), 136.9 (C), 153.6 (C); HRMS (EI) calculated for [C21H24FeO]+ (M+): 348.1171, found 348.1182.
4-[1-(Ferrocenyl)but-3-enyl]phenol (3k): yellow oil; 1H-NMR (300 MHz, CDCl3): 2.53–2.62 (m, 1H, CH2), 2.80–2.89 (m, 1H, CH2), 3.65 (dd, J = 10.5 and 4.7 Hz, 1H, CH), 4.08 (s, 1H, Cp), 4.13–4.10 (m, 7H, Cp), 4.19 (s, 1H, Cp), 4.67 (s, 1H, OH), 4.93–5.04 (m, 2H, =CH2), 5.76–5.77 (m, 1H, =CH), 6.76 (d, J = 8.6 Hz, 2H, Ar), 7.06 (d, J = 8.6 Hz, 2H, Ar); 13C-NMR (75 MHz, CDCl3): 12.7 (CH3), 30.0 (CH2), 47.0 (CH), 66.8 (CH), 66.9 (CH), 67.3 (CH), 67.4 (CH), 68.6 (CH), 94.8 (C), 114.9 (CH), 129.0 (CH), 137.8 (C), 153.6 (C); HRMS (EI) calculated for [C20H20FeO]+ (M+): 332.0858, found 332.0855.
4-(Ferrocenylmethyl)phenol (3l): yellow oil; 1H-NMR (300 MHz, CDCl3): 3.64 (s, 2H, CH2), 4.11 (s, 4H, Cp), 4.15 (s, 5H, Cp), 4.84 (s, 1H, OH), 6.75 (d, J = 8.2 Hz, 2H, Ar), 7.02 (d, J = 8.2 Hz, 2H, Ar); 13C-NMR (75 MHz, CDCl3): 45.2 (CH2), 77.6 (CH), 78.7 (CH), 78.8 (CH), 98.7 (C), 125.1 (CH), 139.6 (CH), 144.0 (C), 163.7 (C); HRMS (EI) calculated for [C17H16FeO]+ (M+): 292.0545, found 292.0524. Ferrocene 3l is a known compound [25].

3.3. Synthesis of Ferrocene Derivative 5

InCl3 (4.4 mg, 0.02 mmol, 10 mol%) was added to a solution of 4-benzylidene-2,6-di-tert-butylcyclohexa-2,5-dienone 4 (58.9 mg, 0.2 mmol) and ferrocene 2 (111.6 mg, 0.6 mmol) in toluene (2 mL). The mixture was stirred at 100 °C for 6 h (disappearance of 4 checked by TLC). Then, the solvent was removed under reduced pressure and the resulting residue was purified by flash chromatography (silica gel, hexanes/ethyl acetate 5:1) to yield ferrocene derivative 5 (76.9 mg, 80%) as a yellow oil; 1H-NMR (300 MHz, CDCl3): 1.44 (s, 18H, CH3), 3.98–3.99 (m, 1H, Cp), 4.01 (s, 5H, Cp), 4.03–4.04 (m, 1H, Cp), 4.16–4.17 (m, 1H, Cp), 4.91 (s, 1H, Cp), 5.06 (s, 1H, CH), 5.09 (s, 1H, OH), 7.04 (s, 2H, Ar), 7.18–7.21 (m, 3H, Ar), 7.26–7.29 (m, 2H, Ar); 13C-NMR (75 MHz, CDCl3): 30.0 (CH3), 34.4 (CH), 51.8 (CH), 67.3 (CH), 67.6 (CH), 68.6 (CH), 68.7 (CH), 68.8 (CH), 92.8 (C), 125.4 (CH), 125.8 (C), 127.9 (CH), 128.6 (CH), 135.1 (C), 135.4 (C), 143.8 (C), 151.9 (C); HRMS (EI) calculated for [C31H36FeO]+ (M+): 480.2110, found 480.2124.

3.4. Cytotoxic Assays

Cell Counting Kit-8 (CCK-8) from Sigma-Aldrich (Madrid, Spain) was used according to the protocol provided by the company. The A2780 and A549 cell lines were used in this preliminary study. First, cell lines were cultured for 7 days in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% (v/v) fetal bovine serum (FBS). Then, cells were seeded into a 96-well flat-bottom culture plate at a cell density of 500–2000 cells/well and incubated for 24 h in the same medium (DMEM/10% FBS). After that, 10 μL of a solution of the corresponding ferrocene derivative at different concentrations were added and the cells were incubated for 72 h. Then, 10 μL of the CCK-8 solution were added to each well of the plate. After 2 h of incubation the absorbance at 450 nm was recorded using a BioTek ELx800 Absorbance Microplate Reader (BioTek, Bad Friedrichshall, Germany). Measurements were performed in triplicate, and each experiment was repeated three times. The IC50 values (μm) were estimated by treatment of the data obtained with the statistical program GraphPad Prism5 (version 5.04).

4. Conclusions

Guided by earlier work from our group, we have developed a convenient synthesis of para-substituted phenol derivatives containing a ferrocenyl moiety. Salient features of our protocol include (i) easy availability of the required starting materials, (ii) synthetically useful yields, and (iii) mild reaction conditions. This C–H bond functionalization of ferrocene relies on the generation of a para-quinone methide intermediate that, activated by Lewis acid complexation, would serve as electrophilic partner in an aromatic electrophilic substitution. Preliminary biological evaluation revealed that some of the ferrocene derivatives available by this protocol display significant cytotoxicity against ovarian and lung cancer cell lines. Further studies aimed at the preparation of new ferrocene derivatives with enhanced antiproliferative properties are being pursued in our laboratory.

Supplementary Materials

Supplementary Materials available online: Copies of 1H- and 13C-NMR spectra, X-Ray crystallography (printcif and checkcif).

Author Contributions

L.A.L. conceived the experiments; S.G.-P. and E.L. designed and performed the experiments; J.B. performed the X-ray crystallographic study; N.d.-l.-S.-A. performed the voltammetry study; L.A.L., J.B. and N.d.-l.-S.-A. wrote the paper.

Acknowledgments

Financial support from the Ministerio de Economía y Competitividad (MINECO, grants CTQ2013-41511-P and CTQ2015-63567-R), Agencia Estatal de Investigación (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) (Grants CTQ2016-76840-R and CTQ2016-75986-P), and Principado de Asturias (grants GRUPIN14-013 and GRUPIN14-006) is gratefully acknowledged. E.L. thanks the Principado de Asturias for a predoctoral grant (Severo Ochoa Program). We also thank J.M. González for interesting discussions.

Conflicts of Interest

The authors declare no conflict of interest.

Appendix A. Crystal Data for Ferrocene Derivative 3a

Crystal Data for C23H20FeO (Mr = 368.24 g/mol): monoclinic, space group P21/n (No. 14), a = 15.662(1) Å, b = 6.0219(3) Å, c = 18.786(1) Å, β = 96.990(6)°, V = 1758.6(2) Å3, Z = 4, T = 299 K, μ(CuKα) = 6.91 mm−1, Dx = 1.391 g/cm3, 8336 measured reflections (3.5° < θ < 69.6°), 3254 independent reflections, 2641 observed reflections (I > 2σ(I)), Rint = 0.045. Final R[F2 > 2σ(F2)] was 0.079 and wR(F2) = 0.255.

Appendix B. Electrochemical Study for Ferrocene Derivative 3a

Compound 3a was studied by cyclic voltammetry (CV). All potentials were referred to a Ag|AgCl|KCl(sat) reference electrode. The ferrocene/ferrocenium couple (Ia/Ic) is clearly observed at a formal potential, E°′ = 0.352 V (Figure A1a). The anodic peak current is higher than the cathodic one indicating that some ferrocenium ions diffuse from the carbon paste electrode to the bulk solution due to the positive charge of the cation. Nucleation at potentials ~0.450 V points out to a dissimilar electrochemical behavior with other structurally related compounds. When the potential is swept up to +0.6 V, a second redox process (IIa/IIc) appears causing the decrease of Ia (Figure A1b). The reduction of ferrocenium species (Ic) remains visible but progressively shifted towards less positive potentials in subsequent scans. Process IIa at about +0.570 V is clear and well-shaped but process IIc at 0.426 V is partially overlapped with Ic. Of note, a notably increase in the non-faradaic current is observed. When the potential is extended to +1.3 V, additional oxidation reactions take place (see the rising anodic current) probably associated to phenolic compounds. Only one reduction process is observed as a result of Ic and IIc overlapping (Figure A1c). The potential of both oxidation and reduction peaks shift to more extreme potentials which indicates that the process is irreversible. The origin of the irreversibility might be the formation of non-conducting products on the electrode surface that hinder the electron transfer. The appearance of resistance affecting the CV shape strongly supports this explanation.
Molecules 23 01335 g0a1 550
Figure A1. Cyclic voltammetry of compound 3a incorporated to the carbon paste electrode in phosphate saline buffer (pH 7.4). Scan rate = 50 mV/s. Potential scan from 0 V to +0.5 V; (a) +0.6 (first scan, black; second scan, blue) and +0.7 (first scan, green; second scan, red) (b) and +1.3 V (c).
Figure A1. Cyclic voltammetry of compound 3a incorporated to the carbon paste electrode in phosphate saline buffer (pH 7.4). Scan rate = 50 mV/s. Potential scan from 0 V to +0.5 V; (a) +0.6 (first scan, black; second scan, blue) and +0.7 (first scan, green; second scan, red) (b) and +1.3 V (c).
Molecules 23 01335 g0a1

References and Notes

  1. Kealy, T.J.; Pauson, P.L. A New Type of Organo-Iron Compound. Nature 1951, 168, 1039–1040. [Google Scholar] [CrossRef]
  2. Miller, S.A.; Tebboth, J.A.; Tremaine, J.F. Dicyclopentadienyliron. J. Chem. Soc. 1952, 632–635. [Google Scholar] [CrossRef]
  3. Štěpnička, P. Ferrocenes: Ligands, Materials and Biomolecules; Wiley: Chichester, UK, 2008. [Google Scholar]
  4. Phillips, E.S. Ferrocenes: Compounds, Properties and Applications; Nova Science Publishers: New York, NY, USA, 2011. [Google Scholar]
  5. Ferrocenes. Homogeneous Catalysis. Organic Synthesis. Materials Science; Togni, A.; Hayashi, T. (Eds.) VCH: Weinheim, Germany, 1995. [Google Scholar]
  6. Chiral Ferrocenes in Asymmetric Catalysis; Dai, L.-X.; Hou, X.-L. (Eds.) Wiley-VCH: Weinheim, Germany, 2009. [Google Scholar]
  7. Astruc, D. Why is Ferrocene so Exceptional? Eur. J. Inorg. Chem. 2017, 1, 6–29. [Google Scholar] [CrossRef]
  8. Arrayás, R.G.; Adrio, J.; Carretero, J.C. Recent Applications of Chiral Ferrocene Ligands in Asymmetric Catalysis. Angew. Chem. Int. Ed. 2006, 45, 7674–7715. [Google Scholar] [CrossRef] [PubMed]
  9. Van Staveren, D.R.; Metzler-Nolte, N. Bioorganometallic Chemistry of Ferrocene. Chem. Rev. 2004, 104, 5931–5986. [Google Scholar] [CrossRef] [PubMed]
  10. Patra, M.; Gasser, G. The medicinal chemistry of ferrocene and its derivatives. Nat. Rev. Chem. 2017, 1, 066. [Google Scholar] [CrossRef]
  11. Dive, D.; Biot, C. Ferroquine as an Oxidative Shock Antimalarial. Curr. Top. Med. Chem. 2014, 14, 1684–1692. [Google Scholar] [CrossRef] [PubMed]
  12. Braga, S.S.; Silva, A.M.S. A New Age for Iron: Antitumoral Ferrocenes. Organometallics 2013, 32, 5626–5639. [Google Scholar] [CrossRef]
  13. Melendez, E. Metallocenes as target specific drugs for cancer treatment. Inorg. Chim. Acta 2012, 393, 36–52. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  14. Ornelas, C. Application of ferrocene and its derivatives in cancer research. New J. Chem. 2011, 35, 1973–1985. [Google Scholar] [CrossRef]
  15. Schatzschneider, U.; Metzler-Nolte, N. New Principles in Medicinal Organometallic Chemistry. Angew. Chem. Int. Ed. 2006, 45, 1504–1507. [Google Scholar] [CrossRef] [PubMed]
  16. Gasser, G.; Ott, I.; Metzler-Nolte, N. Organometallic Anticancer Compounds. J. Med. Chem. 2011, 54, 3–25. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  17. Jaouen, G.; Vessières, A.; Top, S. Ferrocifen type anti cancer drugs. Chem. Soc. Rev. 2015, 44, 8802–8817. [Google Scholar] [CrossRef] [PubMed]
  18. Nguyen, A.; Vessières, A.; Hillard, E.A.; Top, S.; Pigeon, P.; Jaouen, G. Ferrocifens and ferrocifenols as new potential weapons against breast cancer. Chimia 2007, 61, 716–724. [Google Scholar] [CrossRef]
  19. Vessières, A. Metal carbonyl tracers and the ferrocifen family: Two facets of biorganometallic chemistry. J. Organomet. Chem. 2013, 734, 3–16. [Google Scholar] [CrossRef]
  20. Hillard, E.A.; Vessières, A.; Jaouen, G. Ferrocene functionalized endocrine modulators as anticancer agents. Top. Organomet. Chem. 2010, 32, 81–117. [Google Scholar]
  21. Hillard, E.; Vessières, A.; Thouin, L.; Jaouen, G.; Amatore, C. Ferrocene-mediated proton-coupled electron transfer in a series of ferrocifen-type breast-cancer drug candidates. Angew. Chem. Int. Ed. 2006, 45, 285–290. [Google Scholar] [CrossRef] [PubMed]
  22. Hamels, D.; Dansette, P.M.; Hillard, E.A.; Top, S.; Vessières, A.; Herson, P.; Jaouen, G.; Mansuy, D. Ferrocenyl quinone methides as strong antiproliferative agents: Formation by metabolic and chemical oxidation of ferrocenyl phenols. Angew. Chem. Int. Ed. 2009, 48, 9124–9126. [Google Scholar] [CrossRef] [PubMed]
  23. Messina, P.; Labbé, E.; Buriez, O.; Hillard, E.A.; Vessières, A.; Hamels, D.; Top, S.; Jaouen, G.; Frapart, Y.M.; Mansuy, D.; et al. Deciphering the activation sequence of ferrociphenol anticancer drug candidates. Chem. Eur. J. 2012, 18, 6581–6587. [Google Scholar] [CrossRef] [PubMed]
  24. Wang, Y.; Pigeon, P.; Top, S.; McGlinchey, M.J.; Jaouen, G. Organometallic antitumor compounds: ferrocifens as precursors to quinone methides. Angew. Chem. Int. Ed. 2015, 54, 10230–10233. [Google Scholar] [CrossRef] [PubMed]
  25. Hillard, E.; Vessières, A.; Le Bideau, F.; Plazuk, D.; Spera, D.; Huché, M.; Jaouen, G. A Series of Unconjugated Ferrocenyl Phenols: Prospects as Anticancer Agents. Chem. Med. Chem. 2006, 1, 551–559. [Google Scholar] [CrossRef] [PubMed]
  26. Plazuk, D.; Vessières, A.; Le Bideau, F.; Jaouen, G.; Zakrzewski, J. Synthesis of Benzyl- and Benzhydrylferrocenes via Friedel-Crafts Alkylation of Ferrocene. Access to Ferrocenyl Bisphenols with High Affinities for Estrogen Receptors. Tetrahedron Lett. 2004, 45, 5425–5427. [Google Scholar] [CrossRef]
  27. López, E.; Lonzi, G.; López, L.A. Gold-Catalyzed C-H Bond Functionalization of Metallocenes: Synthesis of Densely Functionalized Ferrocene Derivatives. Organometallics 2014, 33, 5924–5927. [Google Scholar] [CrossRef]
  28. López, E.; Borge, J.; López, L.A. Gold-Catalyzed Intermolecular Formal Insertion of Aryldiazo Esters into Cp-H Bonds of Iron and Ruthenium Metallocenes. Chem. Eur. J. 2017, 23, 3091–3097. [Google Scholar] [CrossRef] [PubMed]
  29. López, E.; Suárez, T.; Ballesteros, A.; López, L.A. Gold(I)-Catalyzed Reaction of Ferrocene and Propargylic Esters: Synthesis of Functionalized Ferrocene Derivatives. Eur. J. Inorg. Chem. 2017, 225–228. [Google Scholar] [CrossRef]
  30. González-Pelayo, S.; López, E.; Borge, J.; de-los-Santos-Álvarez, N.; López, L.A. Ferrocene-Decorated Phenol Derivatives by Trapping of ortho-Quinone Methide Intermediates with Ferrocene. Eur. J. Org. Chem. 2018. [Google Scholar] [CrossRef]
  31. The Lewis acid-catalyzed monoalkylation of ferrocene with some benzyl alcohols has been reported. This process takes place in molten ferrocene as solvent at high temperature (176 °C): Abdullah, R.; Chung, K.; Kim, Y.-W.; Hong, I.S. An Efficient Synthesis of Mono-Substituted Benzyl Ferrocene Derivatives. Bull. Korean Chem. Soc. 2015, 36, 32–35. [Google Scholar] [CrossRef]
  32. For a similar methodology for the alkylation of ferrocene, see: Neuse, E.W.; Trifan, D.S. Alkylation of Ferrocene with α-Aryl Alcohols. J. Am. Chem. Soc. 1962, 84, 1850–1856. [Google Scholar]
  33. COD 3000199 Contains the Supplementary Crystallographic Data for This Paper. These Data Can Be Obtained Free of Charge via. See also Supplementary Materials. Available online: http://www.crystallography.net/search.html (accessed on 30 May 2018).
  34. A detailed analysis revealed a non-classical intermolecular interaction of the phenolic OH group with the π cloud of the aromatic ring. For selected examples of aromatic rings as weak hydrogen bond acceptors in crystals, see: Jeffrey, G.A. An Introduction to Hydrogen Bonding; Oxford University Press: New York, NY, USA, 1997. [Google Scholar]
  35. Seminal contribution on Friedel-Crafts acylation of ferrocene: Woodward, R.B.; Rosenblum, M.; Whiting, M.C. A new aromatic system. J. Am. Chem. Soc. 1952, 74, 3458–3459. [Google Scholar] [CrossRef]
  36. For a recent related arylation process involving electron-rich arenes, see: Gao, S.; Xu, X.; Yuan, Z.; Zhou, H.; Yao, H.; Lin, A. 1,6-Addition arylation of para-quinone methides: An approach to unsymmetrical triarylmethanes. Eur. J. Org. Chem. 2016, 2016, 3006–3012. [Google Scholar] [CrossRef]
  37. In this study, NIH 3T3 cells were used as normal control cell line
  38. Wong, Y.F.; Wang, Z.; Sun, J. Chiral phosphoric acid catalyzed asymmetric addition of naphthols to para-quinone methides. Org. Biomol. Chem. 2016, 14, 5751–5754. [Google Scholar] [CrossRef] [PubMed]
  39. McNulty, J.; McLeod, D. A scalable process for the synthesis of (E)-Pterostilbene involving aqueous Wittig olefination chemistry. Tetrahedron Lett. 2013, 54, 6303–6306. [Google Scholar] [CrossRef]
  40. Chu, W.-D.; Zhang, L.-F.; Bao, X.; Zhao, X.-H.; Zeng, C.; Du, J.-Y.; Zhang, G.-B.; Wang, F.-X.; Ma, X.-Y.; Fan, C.-A. Asymmetric catalytic 1,6-conjugate addition/aromatization of para-quinone methides: Enantioselective introduction of functionalized diarylmethine sterogenic centers. Angew. Chem. Int. Ed. 2013, 52, 9229–9233. [Google Scholar] [CrossRef] [PubMed]
Sample Availability: Samples of the compounds are available from the authors.
Molecules 23 01335 g001 550
Figure 1. Phenol-ferrocene conjugates: (a) Ferrocifen family; (b) ferrocene bisphenol derivatives; (c) ortho-substituted ferrocenyl phenols previously developed in our group; (d) para-substituted ferrocenyl phenols reported in this study.
Figure 1. Phenol-ferrocene conjugates: (a) Ferrocifen family; (b) ferrocene bisphenol derivatives; (c) ortho-substituted ferrocenyl phenols previously developed in our group; (d) para-substituted ferrocenyl phenols reported in this study.
Molecules 23 01335 g001
Molecules 23 01335 g002 550
Figure 2. Starting p-hydroxybenzyl alcohols 1 used in this work.
Figure 2. Starting p-hydroxybenzyl alcohols 1 used in this work.
Molecules 23 01335 g002
Molecules 23 01335 sch001 550
Scheme 1. Trapping of p-quinone methides with ferrocene: proof of concept. DCE: dichloroethane.
Scheme 1. Trapping of p-quinone methides with ferrocene: proof of concept. DCE: dichloroethane.
Molecules 23 01335 sch001
Molecules 23 01335 g003 550
Figure 3. X-ray structure of ferrocene derivative 3a. Thermal ellipsoids are drawn at the 30% probability level. Hydrogen atoms are excluded, except those bonded to C7 (H7) and O4 (H4O).
Figure 3. X-ray structure of ferrocene derivative 3a. Thermal ellipsoids are drawn at the 30% probability level. Hydrogen atoms are excluded, except those bonded to C7 (H7) and O4 (H4O).
Molecules 23 01335 g003
Molecules 23 01335 sch002 550
Scheme 2. InCl3-catalyzed reaction of ferrocene and stable p-quinone methide 4.
Scheme 2. InCl3-catalyzed reaction of ferrocene and stable p-quinone methide 4.
Molecules 23 01335 sch002
Molecules 23 01335 sch003 550
Scheme 3. Mechanism for the InCl3-catalyzed reaction of phenol derivatives 1 and ferrocene (2).
Scheme 3. Mechanism for the InCl3-catalyzed reaction of phenol derivatives 1 and ferrocene (2).
Molecules 23 01335 sch003
Table
Table 1. InCl3-catalyzed reaction of p-hydroxybenzyl alcohols 1 and ferrocene (2).
Table 1. InCl3-catalyzed reaction of p-hydroxybenzyl alcohols 1 and ferrocene (2).
Molecules 23 01335 i001
EntrySubstrateR3Yield (%) a
11aC6H53a75
21bo-MeC6H43b76
31cm-MeC6H43c51
41dp-MeC6H43d82
51ep-MeOC6H43e48
61fMe3f62
71gEt3g60
81hn-Bu3h42
91ii-Pr3i62
101jt-Bu3j43
111kAllyl3k44
121lH3l64
a Isolated yield after chromatographic purification.
Table
Table 2. IC50 [μM] values for selected ferrocenyl compounds on different cell lines a.
Table 2. IC50 [μM] values for selected ferrocenyl compounds on different cell lines a.
3a3g
A27801.072.23
A5493.554.89
a Measured after 72 h of culture.

Share and Cite

MDPI and ACS Style

González-Pelayo, S.; López, E.; Borge, J.; De-los-Santos-Álvarez, N.; López, L.A. Trapping para-Quinone Methide Intermediates with Ferrocene: Synthesis and Preliminary Biological Evaluation of New Phenol-Ferrocene Conjugates. Molecules 2018, 23, 1335. https://doi.org/10.3390/molecules23061335

AMA Style

González-Pelayo S, López E, Borge J, De-los-Santos-Álvarez N, López LA. Trapping para-Quinone Methide Intermediates with Ferrocene: Synthesis and Preliminary Biological Evaluation of New Phenol-Ferrocene Conjugates. Molecules. 2018; 23(6):1335. https://doi.org/10.3390/molecules23061335

Chicago/Turabian Style

González-Pelayo, Silvia, Enol López, Javier Borge, Noemí De-los-Santos-Álvarez, and Luis A. López. 2018. "Trapping para-Quinone Methide Intermediates with Ferrocene: Synthesis and Preliminary Biological Evaluation of New Phenol-Ferrocene Conjugates" Molecules 23, no. 6: 1335. https://doi.org/10.3390/molecules23061335

APA Style

González-Pelayo, S., López, E., Borge, J., De-los-Santos-Álvarez, N., & López, L. A. (2018). Trapping para-Quinone Methide Intermediates with Ferrocene: Synthesis and Preliminary Biological Evaluation of New Phenol-Ferrocene Conjugates. Molecules, 23(6), 1335. https://doi.org/10.3390/molecules23061335

Article Metrics

Back to TopTop