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Synthesis, characterization, kinetic investigation and biological evaluation of Re(i) di- and tricarbonyl complexes with tertiary phosphine ligands.

PMID: 31740907
Volume 49 Number 1 7 January 2020 Dalton Pages 1-242 Transactions An international journal of inorganic chemistry rsc.li/dalton ISSN 1477-9226 PAPER Marietjie Schutte-Smith et al. Synthesis, characterization, kinetic investigation and biological evaluation of Re(I ) di- and tricarbonyl complexes with tertiary phosphine ligands Dalton Transactions PAPER Synthesis, characterization, kinetic investigation and biological evaluation of Re( ) di- and I Citethis:DaltonTrans.,2020,49,35 tricarbonyl complexes with tertiary phosphine † ligands VerityLindyGantsho,aMazzarineDotou, bMartaJakubaszek, b,cBrunoGoud, c GillesGasser, bHendrikGideonVisseraandMarietjieSchutte-Smith *a Rhenium(I)di-andtri-carbonylcomplexesoftheformfac-[Re(CO) 3 (L,L’-Bid)X]and[Re(CO) 2 (L,L’-Bid)X 2 ], whereX=aqua(H O),methanol(CH OH),triphenylphosphine(PPh ),1,3,5-triaza-7-phosphaadamantane 2 3 3 (PTA), tricyclohexylphosphine (PCy ) and L,L’-Bid = O,O’ bidentate ligands (tropolone = TropH and 3 3-hydroxyflavone=FlavH)andN,Obidentateligands(8-hydroxyquinoline=QuinH,5,7-chloro-8-hydro- xyquinoline = diCl-QuinH and quinoline-2,4-dicarboxylic acid = QuinH ), were synthesized and un- 2 ambiguouslycharacterizedby1H-,13C-and31P-NMR,IR,UV/Visandmicro-analysis.Thecrystalstructures of four complexes, namely fac-[Re(CO) (QuinH)(H O)]·H O (5), fac-[Re(CO) (Quin)(PPh )] (11), fac- 3 2 2 3 3 [Re(CO) (diCl-Quin)(PPh )](12)and[Re(CO) (Trop)(PPh ) ]·2C H CH (20)wereobtained.Re–Pbonding 3 3 2 32 6 5 3 distancesfor11and12are2.4948(8)and2.4908(8)Å,respectively,indicatingtheeffectoftheelectron- withdrawingsubstituentsofthediCl-Quin−ligand.Thesecond-orderrateconstantsforthesubstitutions of methanol at 25.1 °C in fac-[Re(CO) (L,L’-Bid)(CH OH)] (L,L’-Bid = Trop, Flav and QuinH) type com- 3 3 plexesbydifferententeringphosphineligands(PPh ,PCy ,andPTA)variedbetween7.23(7)×10−5and 3 3 1.32(3) × 10−3 M−1 s−1 and were found to depend on the coordinated bidentate ligand (in general k 1 (QuinH) < k (Trop) < k (Flav)). The toxicityof fac-[Re(CO) (QuinH)(PTA)], fac-[Re(CO) (Trop)(PTA)], fac- 1 1 3 3 [Re(CO) (Trop)(PPh )]andfac-[Re(CO) (Flav)(PPh )]onthecervicalcancerHeLaandepithelialRPE-1cell 3 3 3 3 Received14thOctober2019, lines wasthen evaluated. Complex fac-[Re(CO) (Flav)(PPh )] (16) and fac-[Re(CO) (Trop)(PPh )] (13) dis- 3 3 3 3 Accepted7thNovember2019 playedthehighestcytotoxicitywithIC valuesof12.21±0.17µMand13.35±0.94µM,respectivelyin 50 DOI:10.1039/c9dt04025k HeLa cells. Interestingly, a small selectivity towards cancer over non-cancerous cells was observed for rsc.li/dalton thesecompounds(IC 50 =18.41±3.16µMand>25µMinRPE-1cells). Introduction used to mimic Technetium-99m, the widely used PET radio isotope.1–10Ontopofthis,rhenium(I)complexesareseriously Rhenium(I) complexes increasingly exhibit interesting pro- investigated by seveal groups for its chemotherapeutic pertiesinthefightagainstcancer,especiallyduetotheirmul- properties.1–5 The use of phosphine ligands coordinated to tiple potential applications. Rhenium-186 is investigated for technetium and rhenium isthe driving force for imaging and potential application as a radiation therapy agent, while therapy in nuclear medicine.6–11 Due to its π-acceptor and σ- Rhenium-188 has properties that could be exploited in PET. donor properties, phosphines can be used as reducing agents − Furthermore, it has been shown that cold rhenium can be towards the [MO ] (M = Tc, Re) salts, as well as chelating 4 agents to stabilize the metal in intermediate to low oxidation states.12–14 The chemistry has been well-developed and two cationic complexes anchored by phosphines have been aUniversityoftheFreeState,DepartmentofChemistry,NelsonMandelaDrive, introduced for myocardial perfusion imaging purposes: Bloemfontein,SouthAfrica.E-mail:schuttem@ufs.ac.za bChimieParisTech,PSLUniversity,CNRS,InstituteofChemistryforLifeandHealth [99mTcO 2 (P 53 ) 2 ]+(Myoview)and[99mTc-Q 12 ]+(TechneScanQ 12 ).15,16 Sciences,LaboratoryforInorganicChemicalBiology,Paris,France The use of fac-[M(CO) (H O) ]+ (M = Tc, Re) for the pro- 3 2 3 cInstitutCurie,PSLUniversity,CNRSUMR144,Paris,France ductionofradiopharmaceuticalsfortherapeutic(Re)anddiag- †Electronic supplementary information (ESI) available: All single crystal X-ray nostic (Tc) purposes is well-established.8,17 The three water data.CCDC1947424(5),1950181(11),1950222(12)and1947425(20).ForESI ligands can easily be replaced with innovative ligand systems. and crystallographic data in CIF or other electronic format see DOI: 10.1039/ c9dt04025k This opens an opportunity for the formation of metal com- Thisjournalis©TheRoyalSocietyofChemistry2020 DaltonTrans.,2020,49,35–46 | 35 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online View Journal | View Issue Paper DaltonTransactions plexes with target tissue specificity and high in vivo stability, Bid = O,O′ bidentate ligands (tropolone = TropH and 3-hydro- amongst other factors that are necessary in radiophamarma- xyflavone=FlavH)andN,Obidentateligands(8-hydroxyquino- ceuticaldesign.18 line=QuinH,5,7-chloro-8-hydroxyquinoline=diCl-QuinHand Preparing water-soluble, organometallic transition metal quinoline-2,4-dicarboxylic acid = QuinH ). The substitution 2 complexes have received attention for applications in various kinetics of selected compounds and their cell toxicity is also areasofresearch.19–21Watersolubilityofmetalcomplexesisa investigated in order to see if a parallel between substitution desired feature in radiopharmacy and nuclear medicine reactivity and cell toxicity can be drawn. The latter is of par- because such compounds have better in vivo characteristics ticular interest since there is a surge in the interest of using (e.g.,clearanceofthedrugfromthebodyisfaster).Significant rhenium complexes as anticancerorantibacterial compounds progress has been made over the years on synthesizing orasphotodynamictherapyphotosensitizers.32–41 water-soluble complexes for diagnostic and therapeutic purposes.22,23 Water solubility is generally achieved by using a variety of water-soluble ligands. 1,3,5-Triaza-7-phospha- Results and discussion adamantane(PTA)isoneofthemonodentatephosphineligands Complexsynthesis used in this studyand was specifically chosen due to its high water solubility. Becauseofthewatersolubility, high-oxidative The preparation of the rhenium starting synthon, fac- stability and low steric requirements of PTA, investigations [NEt 4 ] 2 [Re(CO) 3 (Br) 3 ] (1), was performed according to a pro- have confirmed that ligand substitution with fac- cedure described by Alberto et al.4 The CO stretching fre- [NEt ] [ReBr (CO) ] is relatively easy and can be useful in the quencyof1featuresasignalat1846and1996cm −1usingKBr 42 3 3 designofinnovativeradiopharmaceuticals.24 asreference. Byusingthe‘2+1’approach,thethreelabileaqualigands ThroughtheadditionofAgNO 3 inacidicwater(pH2.2),the onfac-[M(CO) 3 (H 2 O) 3 ]+aresubstitutedbyaN,O/O,O′-bidentate three Br − of 1 could be replaced with H 2 O molecules forming ligand and a monodentate ligand.25 For this study, the phos- fac-[Re(CO) 3 (H 2 O) 3 ]+ (2).4 The carbonyl ligands on 2 are phineligandiscoordinatedtothemetalcentreasamonoden- stronglyboundtothemetalcentre,whilethethreewatermole- tate ligand and,in turn,causesthe carbonylligand transto it cules are labile and are effortlessly displaced byother ligands to become more labile, allowing it to be substituted by a (Scheme1).5 second phosphine ligand, forming a Re(I) dicarbonyl species, Complexes 3–7 were obtained by adding various bidentate whichwillpotentiallydisplayevenbetterinvivocharacteristics ligandsto 2, resulting intwo ofthe H 2 O ligandsbeing substi- thantheRe(I)tricarbonylcomplex.Thisapproachisknownas tuted to produce fac-[Re(CO) 3 (L,L′-Bid)(H 2 O)] type complexes the‘2+1+1’approach.26 where L,L′-Bidisthe bidentateligand. Thekineticbehaviorof ReactivitystudiesofRe(I)tricarbonylcomplexeshaveadual complexes 5–7 upon the introduction of a phosphine mono- purpose. Firstly, it serves as a model for Tc(I) complexes and dentate ligand was performed in methanol. A relatively quick secondly it assists in predicting in vivo behavior. It is well- substitutionoccursbetweentheH 2 Omoleculeon5–7andthe known that metal complexes undergo changes when they methanol,producing8–10(Scheme1). come into contact with nucleophiles like thiocyanate in The carbonyl ligand trans to the coordinated phosphine blood.27–29 Information on the type of mechanism (dissocia- ligand can be substituted with a second phosphine through tive vs. associative) provides important information in the the addition of excess phosphine ligand in toluene as solvent design of new complexes, as recently discussed in detail.30 at 80 °C to produce 20 (Scheme 2). All complexes could be This analytical tool can give sufficient information about the characterizedbyinfraredaswellas1H,13Cand31PNMRspec- invivometalstabilityandreactivity,aswellastherateofdistri- trometry and micro-analysis of which characteristic spectro- bution and clearance from the body. For that reason, this scopic details are given in the Experimental section study aims at elucidating the mechanism of substitution for (Scheme3). Re(I) tricarbonyl complexes with various phosphine ligands, enablinghencefurthercharacterizationofpotentialradiophar- maceuticals. The formation of the Re(I) dicarbonyl complexes occurs at high temperatures, thus limiting kinetic investi- gations for the substitution of the carbonyl due to equipment restrictions, as reported by Manicum et al.31 They concluded that the removal of a CO ligand from Re(I) tricarbonyl com- plexes, similar to those used in this study, only takes place at hightemperatures. We report here the synthesis of a range of Re(I) di- and tri- carbonyl complexes of the form fac-[Re(CO) (L,L′-Bid)X] and 3 [Re(CO) (L,L′-Bid)X ], where X = aqua (H O), methanol 2 2 2 (CH 3 OH), triphenylphosphine (PPh 3 ), 1,3,5-triaza-7-phospha- Scheme1 Synthesis of fac-[Re(CO) 3 (H 2 O) 3 ]+ (2), fac-[Re(CO) 3 (L,L’-Bid) adamantane(PTA)andtricyclohexylphosphine(PCy 3 )andL,L′- (H 2 O)](3–7)andfac-[Re(CO) 3 (L,L’-Bid)(CH 3 OH)](8–10). 36 | DaltonTrans.,2020,49,35–46 Thisjournalis©TheRoyalSocietyofChemistry2020 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online DaltonTransactions Paper Scheme2 Synthesis of fac-[Re(CO) (L,L’-Bid)(P)] (11–19) and [Re 3 (CO) (Trop)(PPh ) ]·2CH (20). 2 32 7 8 Scheme3 Representationofthecomplexessynthesized. X-Raycrystallography Detailed structural information of these complexes were obtained from crystallographic analyses. Slow evaporation of thesolvent (water/methanol/toluene)fromcomplexes5,11,12 and 20 resulted in single crystals, yielding the subsequent molecular structures (Fig. 1). All the crystallographic data, including anisotropic displacement parameters, all bond dis- tancesandangles,atomicandhydrogencoordinatesaregiven inthe ESI,†whileselected bonddistancesand anglesare pre- sented in Table 1. Hydrogen bonding interaction and π–π Fig.1 Molecularrepresentationofthecrystalstructuresof5,11,12and 20.Somehydrogenatomsareomittedforclarity.Ellipsoidsaredrawnat interactiondataisalsopresentedintheESI.† 50%probability. Complex 5 crystalized in the monoclinic C2/c space group. The Re(I) metal centre is coordinated to three facial carbonyl ligands, a quinoline-2-carboxylato-4-carboxylic acid bidentate Table1 Asummaryofselectedbonddistances(Å)andbiteangles(°)of ligandintheequatorialplaneandanaqualigandintheaxial thecrystalstructuresof5,11,12and20 position. Two Re(I) units and two solvent water molecules are foundintheasymmetricunit.Asignificantdeviationfromthe Complex predicted 90° angles for octahedral complexes is seen in the (5)a (11) (12) (20) bond angles of 5, making the complex distorted. This is evident from the following angles: O04–Re1–O08 (81.76(10)°, Re1–C01(Å) 1.915(3)/1.912(4) 1.920(3) 1.924(3) 1.883(3) 76.89(9)°) and N1–Re1–O04 (75.31(9)°, 75.26(9)°). All other R R e e 1 1 – – C C 0 0 3 2 ( ( Å Å ) ) 1 1 . . 9 9 1 0 1 0 ( ( 4 4 ) ) / / 1 1 . . 9 9 0 0 8 1 ( ( 4 4 ) ) 1 1 . . 9 9 5 1 4 2 ( ( 3 3 ) ) 1 1 . . 9 9 5 0 2 8 ( ( 3 3 ) ) — 1.887(3) bond lengths and angles for 5 compare well with those of Re1–X(Å) 2.228(3)/2.204(3) 2.189(3) 2.184(2) 2.158(2) similar structures.42,43 The structure of 5 is stabilized by one Re1–O04(Å) 2.145(2)/2.142(2) 2.130(2) 2.149(2) 2.155(2) intermolecular π–π interaction between the two rings of the R R e e 1 1 – – P P 2 1 ( ( Å Å ) ) — — — 2.4948(8) — 2.4908(8) 2 2 . . 4 4 2 3 3 0 9 2 ( ( 8 8 ) ) bidentateligandandfourteeninter-andintramolecularhydro- Re1–O8(Å) 2.203(3)/2.185(3) — — — gen interactions. The bidentate ligand (plane through N1, X–Re1–Y(°) 75.31(9)/75.26(9) 76.08(9) 75.62(8) 72.96(7) O04,C04,C05,C06,C07,C08,C09,C10,C12,C13,C14)twists X = N1 (5, 11, 12) and O03 (20). aTwo molecules in the asymmetric significantlyfromtheplanethroughRe1,C02,O02,C03,O03, unit(moleculeA/moleculeB). N1,O04 withadihedral angleof 16.41(9)° and27.99(7)° for A andBrespectively(Fig.2). The basic structures of 11 and 12 are verysimilar. In both, A slight distortion in the octahedral geometry can be seen the Re(I) metal centre is coordinated to three facial carbonyl in both structures with bond angles C01–Re1–O04 (97.38(10)°) ligands, an 8-hydroxyquinolinato (11)/5,7-dichloro-8-hydroxy- andC02–Re1–N1(98.00(10)°)in11andC01–Re1–O04(99.52(10)°) quinolinato(12)bidentateligandintheequatorialplaneanda and C02–Re1–N1 (96.96(11)°) in 12 which deviates sig- triphenylphosphineligandintheaxialposition. nificantlyfromthepredictedninetydegrees.TheRe–CObond Thisjournalis©TheRoyalSocietyofChemistry2020 DaltonTrans.,2020,49,35–46 | 37 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online Paper DaltonTransactions bite angle(O04–Re1–O03) of72.96(7)° which is comparable to similar structures.50 Complex 20 contains one weak (3.882(6) Å)intramolecularπ–πinteractionbetweenthebidentateligand and the phenyl ring from the monodentate triphenyl- phosphine ligand. The tropolonato bidentate ligand bends slightlytowardsoneofthePPh ligandswithadihedralangle 3 of 8.46(7)° between the planes through Re1, C01, O01, C02, O02,O03,O04andO03,O04,C1,C2,C3,C4,C5,C6,C7. The Re–CO bond distances in the equatorial plane (Re1– C01 and Re1–C02) of 11, 12 and 5 are all considered to be within the predicted range of ∼1.90 Å for similar complexes.44,45 The Re–CO bond distances in the axial plane Fig.2 Illustration of the out-of-plane bending and/or twisting of the (Re1–C03)of11,12and5areslightlylongerthantheequator- bidentateligandin5(moleculeAandB),11,12and20. ialRe–CObonddistances,whichispossiblyattributabletothe transinfluenceoftheligandoppositetoit.51TheRe–CObond distances in the equatorial plane of 20 are slightly shorter distances are comparable to other similar structures, with (1.883(3) and 1.887(3) Å, respectively). The Re1–N1 bond dis- Re1–C02, Re1–C01 and Re1–C03 reported as 1.917(3) Å, tancesare consideredtobewithinnormalrangeof∼2.20Å.42 1.910(3) Å and 1.952(3) Å, respectively for 11, and 1.924(3) Å, The Re1–O04 bond distances of the functionalized hydroxy- 1.908(3) Å and 1.952(3) Å, respectively for 12, all of which are quinolinatoligands12and5areslightlylongercomparedtothat considered within normal range.44–46 Both bidentate ligands, ofthestandard8-hydroxyquinolinatoin11butarestillwithin 8-hydroxyquinolinato and 5,7-dichloro-8-hydroxyquinolinato range of similar structures. The Re1–O03 and Re1–O04 bond show a minimally constrained bite angle (O04–Re1–N1) of distances of 20, 2.1578(19) and 2.1548(18) Å, respectively, are 76.05(8)° and 75.62(8)°, respectively, which is comparable to slightlylongerthanthatofsimilarstructures,rangingbetween similarstructures.47 2.12–2.14Å.46TheRe1–P1of11and12fallwithinthenormal Both crystal structures 11 and 12 are stabilized by four π–π rangeof∼2.50ÅbuttheRe1–P1andRe1–P2of20areslightly interactions, three ofwhich are intramolecularand oneweaker shorter, and range from ∼2.42 Å to ∼2.43 Å, which could be intermolecular in nature (see ESI†). Interactions between the attributed to the stabilizing effect of the phosphine ligands.47 central ring of the bidentate ligand and the phenyl ring from The bite angles of the quinolone ligands in 5, 11 and 12 are the triphenylphosphine ligand contribute to the π-interactions well within range of similar structures (75.26(9)°–76.08(9)°) as of these molecules. Two hydrogen bonding interactions are well asthat ofthe tropolonato ligand20(72.96(7)°).50 The out present in 11, one intermolecular (H33–O04) and the other ofplanebendofthebidentateligandof11,12and20ismuch intramolecular (H22–O04), contributing to the packing of the smallercomparedtothatof5,evidentfromFig.2. molecule. Contrary to 11, structure 12 display three additional Kineticexperiments inter- and intramolecular hydrogen interactions as a result of the chloride atomsattached tothe bidentateligand. Structures Thesubstitutionkineticsof8,9and10withPPh 3 ,PCy 3 andPTA 11 and 12 have a slight out of plane bend of the bidentate asenteringligands,respectivelywerestudiedatvarioustempera- ligand (N1, O04, C1, C2, C3, C4, C5, C6, C7, C8, C9) from the tures.Allreactionswereperformedunderpseudofirst-ordercon- planethroughRe1,C01,O01,C02,O02,N1,O04withadihedral ditionswiththeenteringligandinlargeexcessineachcase.The angleof6.2(2)°and 9.25(7)°for11and12respectively(Fig.2). rates of the reactions increase linearly with an increase of the 20 crystalized in the triclinic P ˉ 1 space group. The Re(I) concentrationoftheenteringligandandtimeversusabsorbance metalcentreiscoordinatedtotwocarbonylligands,atropolo- spectraofallthereactionsstudiedrevealsisosbesticpoints,indi- nato bidentate ligand in the equatorial plane and two tri- cating one-step reactions. Based on this a general mechanism, phenylphosphineligandscoordinatedasmonodentateligands illustratedinScheme4,couldbepresented. in the axial position. Two toluene solvent molecules complete Thereactionratecanbedenotedas: theasymmetricunit;oneofthesearedisorderedovertwoposi- Rate¼k 1 ½ReðL;L′BidÞ(cid:2)½L(cid:2)(cid:3)k(cid:3)1 ½ReðL;L′BidÞðLÞ(cid:2) tions in a 41.4:58.6 ratio. There is a slight distortion in the octahedral geometry around the rhenium atom that can be where k 1 represents the forward reaction and k−1 the reverse seen in the bond angles C01–Re1–O04 (98.15(9)°) and C02– reaction. [Re(L,L′-Bid)] is fac-[Re(L,L′-Bid)(CO) 3 (CH 3 OH)] and Re1–O03 (102.93(10)°), which deviates significantly from the predicted 90°. Re–CO bond distances are comparable to others, withRe1–C01and Re1–C02 reported as1.883(3) Åand 1.887(3)Årespectively,allofwhichareconsiderednormal.48,49 The Re1–O03 and Re1–O04 distance of 2.1578(19) Å and 2.1548(18) Å, respectively compare well with similar struc- Scheme4 Schematic representation of the predicted mechanism for tures.50Thetropolonatoligandshowsaminimallyconstrained themethanolsubstitutionreactionoffac-[Re(CO) (L,L’-Bid)(CH OH)]. 3 3 38 | DaltonTrans.,2020,49,35–46 Thisjournalis©TheRoyalSocietyofChemistry2020 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online DaltonTransactions Paper Table2 Asummaryofthekineticdataforthesubstitutionkineticsof8, 9and10withPTA,PPh andPCy atdifferenttemperatures 3 3 fac-[Re(CO) (Trop)(CH OH)]andPTA 3 3 15.1°C 25.1°C 35.1°C 45.1°C k (M−1s−1) 0.108(4) 0.331(6) 0.934(2) 2.423(6) 1 K 1 03 (M k− −1 1 ( ) s−1) 1 0 5 .0 1 7 3 1 ( 4 5 ( 9 8 ) ) 1 0 5 .2 6 1 8 1 ( ( 3 3 6 ) ) 1 0 5 .6 3 0 8 7 ( ( 2 8 1 ) ) 1 1 4 .6 4 8 2 ( ( 4 3 ) 4) 1 ‡ ΔH ðkJmol(cid:3)1Þ 72(2) ‡ ðk1 Þ ΔS ðJK(cid:3)1mol(cid:3)1Þ −12(6) ðk1 Þ fac-[Re(CO) (Trop)(CH OH)]andPPh 3 3 3 15.1°C 25.1°C 35.1°C 45.1°C k (M−1s−1) 0.1476(8) 0.4189(3) 1.1401(6) 3.3082(4) 1 K 1 03 (M k− −1 1 ( ) s−1) 5 0 7 .0 9 2 7 5 ( 4 3 6 3 ( ) 5) 5 0 7 .0 9 7 3 2 ( 3 5 ( 6 7 ) ) 6 0 0 .1 3 8 2 9 ( ( 6 2 4 ) ) 7 0 6 .4 2 3 2 4 ( ( 1 6 0 ) 5) 1 ‡ ΔH ðkJmol(cid:3)1Þ 70(1) ‡ ðk1 Þ ΔS ðJK(cid:3)1mol(cid:3)1Þ −16(5) ðk1 Þ Fig.3 (a)AtypicalUV/Visspectrumofthemethanolsubstitutionreac- fac-[Re(CO) 3 (Trop)(CH 3 OH)]andPCy 3 tion between fac-[Re(CO) (Flav)(CH OH)] and PCy at 25.1 °C. [Re] = 2 × 10−4 M,[PCy ] =0.01 3 0 M. (b) A 3 fit ofabsorban 3 ce vs. time for this 15.1°C 25.1°C 35.1°C 45.1°C 3 t r r e e e m a a c c p t t i i e o o r n n at a u b t re e λ s tw . = e 4 e 3 n 0n fa m c- , [ Δ Re t ( = CO 28 ) 3 . ( 2 Fl s a . v) ( ( c C ) H A 3 O pl H ot )] of an k o d bs P vs C . y [ 3 PC a y t 3 ] v fo ar r io th u e s 1 K k 1 0 1 3 ( ( M ‡ M k− − −1 1 1 ( ) s s − − 1 1 ) ) 0 0 1 . . 4 0 0 7 0 1 ( 2 9 5 8 4 ) 6 (7 (2 ) ) 0 0 1 . . 5 0 0 9 0 4 ( 6 2 2 7 7 ) 9 (6 9 ) (8) 1 0 0 7 . . 0 0 7 1 9 ( 7 7 8 3 9 ) ( ( 8 3 ) ) 2 0 0 0 . . 0 2 3 4 1 ( 3 5 4 7 ( ) 4 ( ) 5) ΔH ðkJmol(cid:3)1Þ 67(2) ‡ ðk1 Þ ΔS ðJK(cid:3)1mol(cid:3)1Þ −61(7) [Re(L,L′-Bid)(L)]istheproductofsubstitution,fac-[Re(L,L′-Bid) ðk1 Þ fac-[Re(CO) (Flav)(CH OH)]andPTA (CO) (L)]andListheenteringphosphineligand. 3 3 3 Thethreecomplexeswere selectedtocompare theeffectof 5.1°C 15.1°C 25.1°C 35.1°C bidentate donor ligands on reaction rates. PicoH (pyridine-2- k (M−1s−1) 0.524(3) 1.0102(6) 2.1207(6) 4.9872(4) c la a t r o b - o 4- x c y a la rb to o - x 4 y - l c i a c rb a o c x id yl ) ic d a is c p id la ) y an si d m Q il u a i r n it H y ( i q n u s in tr o u l c in tu e r -2 e -c a a n r d bo f x o y r - 1 K 1 0 1 3 ( ‡ M k− −1 1 ( ) s−1) 3 0 7 .1 0 4 1 1 ( 6 2 ( 1 8 ) ) 4 0 2 .2 6 3 2 7 ( ( 3 2 6 ) ) 4 0 9 .4 4 2 3 9 ( ( 4 4 6 ) ) 5 0 8 .8 4 5 7 3 ( ( 5 8 4 ) ) thisreason,werethoughttodisplaycompellingkineticeffects ΔH ‡ ðk1 Þ ðkJmol(cid:3)1Þ 51(3) suitable for comparison. The kinetic data for PicoH was pre- ΔS ðJK(cid:3)1mol (cid:3)1Þ −67(12) ðk1 Þ viouslyinvestigatedbySchutteetal.andwillbereferredtofor comparison purposes.45 Fig. 3a illustrates a typical UV/Vis fac-[Re(CO) 3 (Flav)(CH 3 OH)]andPPh 3 spectrum for the reaction between the phosphine ligand and 5.1°C 15.1°C 25.1°C 35.1°C k (M−1s−1) 1.373(9) 3.709(6) 10.758(5) 29.972(4) 1 K 1 03 (M k− −1 1 ( ) s−1) 0 3 . 7 0 9 3 2 6 ( 2 6 5 7 ( ) 6) 3 0 8 .0 7 9 1 5 ( 8 2 2 1 ( ) 5) 4 0 5 .2 1 3 2 8 ( 4 1 ( 5 8 ) ) 4 0 5 .6 6 5 2 7 ( 8 5 ( 6 7 ) ) 1 ‡ ΔH ðkJmol(cid:3)1Þ 56(4) ‡ ðk1 Þ ΔS ðJK(cid:3)1mol(cid:3)1Þ −39(13) ðk1 Þ fac-[Re(CO) (Flav)(CH OH)]andPCy 3 3 3 15.1°C 25.1°C 35.1°C 45.1°C k (M−1s−1) 1.246(2) 2.153(4) 4.524(7) 8.997(2) 1 K 1 03 (M k− −1 1 ( ) s−1) 0 2 . 1 5 0 9 1 3 ( ( 2 7 5 ) ) 1 1 . 6 3 3 2 1 ( ( 3 3 ) 7) 2 1 . 7 5 6 6 7 ( ( 8 5 ) 5) 4 1 . 8 9 3 1 2 ( ( 5 1 ) 8) 1 ‡ ΔH ðkJmol(cid:3)1Þ 49(2) ‡ ðk1 Þ ΔS ðJK(cid:3)1mol(cid:3)1Þ −74(9) ðk1 Þ fac-[Re(CO) (QuinH)(CH OH)]andPTA 3 3 25.1°C Fig.4 (a)Aplotofligandconcentrationvs.k obs forfac-[Re(CO) 3 (Trop) 103k (M−1s−1) 8.209(8) v (C s. H k 3 o O bs H f ) o ], r [ f R a e c ] -[ = Re 2 (C × O 10 ) 3 − (F 4 la M v) a (C t2 H 5 3 . O 1 H °C )] . , ( [ b R ) e A ]= pl 2 ot × o 1 f 0 l − ig 4 a M nd at co 25 n . c 1 e ° n C tr . ation 1 K 0 1 3 (M k 1 − −1 1 ( ) s−1) 2 0 8 .0 1 2 ( 9 3 ( 9 4 ) ) Thisjournalis©TheRoyalSocietyofChemistry2020 DaltonTrans.,2020,49,35–46 | 39 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online Paper DaltonTransactions fac-[Re(Flav)(CO) (CH OH)] in methanol. The time vs. absor- electron delocalization. From the abovementioned findings, it 3 3 bance datawas fitted to single exponentials (Fig. 3b) confirm- can be concluded that the influence of the bidentate ligands ingfirst-orderbehavior.Thek valueswereobtainedforeach areasfollows:k (PicoH)<k (QuinH)<k (Trop)<k (Flav). obs 1 1 1 1 complexandplottedagainstligandconcentrationsatdifferent From this data it can be observed that the first order rate temperatures(Fig.3c).Fig.4(aandb)illustratesasummaryof constants, k , generally decrease for PPh > DMAP > PTA > 1 3 the reaction plots of fac-[Re(CO) (Trop)(CH OH)] (9) and fac- PCy > Py, however, this does not coincide for an associative 3 3 3 [Re(CO) (Flav)(CH OH)](10)withdifferententeringligands. activated mechanism since the pK of PCy (9.7)52 is larger 3 3 a 3 Three neutral Re(I) tricarbonyl complexes were selected for than that of DMAP (9.6),53 PPh 3 (7.64),54 PTA (6.0)55 and Py thisstudy,oneofwhichcontainedaN,O-bidentateligandand (5.25).56 Interestingly Schutte et al. did not observe a trend in twowithO,O′-bidentateligands.Anaxialmethanolligandwas k values for the monodentate ligands and pK either, which 1 a available for substitution by monodentate entering ligands. further suggests that association does not play an important Theratedataforthemethanolsubstitutionreactionsbetween roleinthereactionmechanism. the three complexes with various phosphine ligands (PTA, Negative ΔS ‡ values normally point towards associative be- PPh and PCy ) are summarized in Table 2. The kinetic data havior, but our recent work on the substitution reactions of 3 3 obtainedbySchutteetal.45isincludedinTable3forcompari- fac-[Re(Trop)(CO) (CH OH)] under various pressures, strongly 3 3 sonreasons. suggests dissociative behavior for the substitution of methanol The values of the second order rate constants, k , for the byvariousenteringligands.30 Here, rather large positive values 1 complex with a N,O-bidentate ligand is much smaller than fortheactivationvolumes(+10–14cm3mol −1)ofreactionswith thosewithO,O′-bidentateligands.Thisisnotexplainablewith variousenteringligandsunderdifferentpressureswasreported, the current knowledge at hand. The results obtained are con- pointingstronglytoadissociativeactivatedmechanism. gruent with those obtained by Schutte et al.44,45 who further TheO,O′bidentateligands(TropandFlav)seemtoprovide established that k fac-[Re(Flav)(CO) (OH )] > k fac-[Re more stable substitution products. In terms of the Flav com- 1 3 2 1 (TropBr )(CO) (OH )], and attributed it to the electron with- plexes fac-[Re(CO) (Flav)(PTA)] had the highest value (4943(46) 3 3 2 3 drawing effects of the bromido substituents on the TropBr M −1). The stability constants, K , of the O,O′ bidentates (Trop 3 1 backbone.Thisisinaccordancewiththevaluesobtainedfrom andFlav-complexes)areingeneralhigherthanthosereported thisstudywherethek valueobtainedforthereactionbetween fortheN,O-bidentateligands,whichinturnsfavoursthemfor 1 fac-[Re(Flav)(CO) (CH OH)] and PTA is more than 6 times useinfuturedrugdesign. 3 3 faster than the reaction between fac-[Re(Trop)(CO) (CH OH)] 3 3 Cytotoxicity andPTA.Thek valueobtainedforfac-[Re(Flav)(CO) (CH OH)] 1 3 3 and PPh is approximately 25 times faster than the similar ColdReorganometalliccompoundshavebeenwidelyexplored 3 reaction with fac-[Re(Trop)(CO) (CH OH)] and 5 times faster as luminescent probes for cell imaging.57–62 In addition, as 3 3 than fac-[Re(Flav)(CO) (CH OH)] and PTA. From the results mentioned in the introduction, several rhenium compounds 3 3 obtained in this study, as well as those obtained by Schutte have been established which display cytotoxicity equal to or et al.44,45 k (QuinH) > k (PicoH), this could be attributed to exceedingthatofcisplatin.32,37,38,63–76 1 1 Table3 Rateconstantsofmethanolsubstitutionreactionsbetweenvariousfac-[Re(CO) (L,L’-Bid)(CH OH)]complexeswithdifferentmonodentate 3 3 ligandsat25.1°C [Re(Trop)] [Re(Flav)] [Re(QuinH)] [Re(PicoH)] PTA k (M−1s−1) 0.331(6) 2.1207(6) 8.209(8)×10−3 k K 1 −1 (M (s− − 1 1 ) ) 1 2 5 .1 6 1 8 ( ( 3 3 ) 6 × ) 10−4 4 4 9 .2 4 9 3 ( ( 4 4 ) 6 × ) 10−4 2 2 8 .9 1 ( ( 4 3 ) 9 × ) 10−5 1 PPh k (M−1s−1) 0.4189(3) 10.758(5) 3 K k 1 −1 (M (s− − 1 1 ) ) 5 7 7 .2 9 3 3 ( ( 7 5 ) 6 × ) 10−5 4 2 5 .3 1 8 2 4 ( ( 1 8 5 ) ) ×10−4 1 PCy k (M−1s−1) 0.006799(8) 2.153(4) 3 K k 1 −1 (M (s− − 1 1 ) ) 1 4 5 .2 9 7 ( ( 2 6 ) )×10−5 1 1 6 .3 1 2 8 ( ( 3 3 ) 7 × ) 10−3 1 Pya k (M−1s−1) 1.38(8) 0.00331(2) 0.00164(8) K k 1 −1 (M (s− − 1 1 ) ) 4 0 6 .3 0 0 0 ( ( 1 1 ) 0 × 0) 10−3 6 0 5 .0 ( 5 9 ( ) 7)×10−3 2 0 1 .0 ( 3 2 ( ) 2)×10−3 1 DMAPa k (M−1s−1) 5.10(2) 0.00652(9) 0.00321(4) K k 1 −1 (M (s− − 1 1 ) ) 3 0 2 .1 0 6 0 (4 0 ) (8 × 0 1 0 0 0 − ) 3 2 0 6 .0 0 2 ( ( 3 3 0 ) ) ×10−3 2 0 9 .1 ( 1 3 ( ) 1)×10−3 1 aKineticdataobtainedbySchutteetal. 40 | DaltonTrans.,2020,49,35–46 Thisjournalis©TheRoyalSocietyofChemistry2020 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online DaltonTransactions Paper Table4 ObtainedIC values(inµM)after48h Quin complexes and in general also more cytotoxic. This 50 initial data would suggest that cytotoxicity might not comple- HeLa RPE-1 tely be ascribed to the formation of adducts withDNA or pro- fac-[Re(CO) (Quin)(PTA)] 44.31±1.44 >100 teinsforthesetypesofcomplexes. 3 fac-[Re(CO) (Trop)(PTA)] >100 >100 3 fac-[Re(CO) (Trop)(PPh )] 13.35±0.94 >25 3 3 fac-[Re(CO) (Flav)(PPh )] 12.21±0.17 18.41±3.16 3 3 Experimental section Cisplatin 8.02±0.57 39.07±0.45 Generalproceduresandinstrumentation The UV/vis spectra were performed on a Varian Cary 50 Conc Rhenium complexes containing water or methanol are UV/Visible Spectrophotometer, equipped with a Julabo prone to substitution (as illustrated by this study). The cyto- F-12 mV temperature cell regulator (accurate to 0.1 °C) in a toxicityobserved is assumed to be related to the formation of 1.000±0.001cmquartzcuvettecell. adducts with DNA or proteins. Due to poor solubility, in this The infrared spectra of the complexes were recorded on a study, cytotoxicity was investigated only for the fac-[Re Bruker Tensor 27 Standard System spectrophotometer with a (CO) (QuinH)(PTA)], fac-[Re(CO) (Trop)(PTA)], fac-[Re 3 3 laser range of 4000–370 cm −1, coupled to a computer. The IR (CO) (Trop)(PPh )] and fac-[Re(CO) (Flav)(PPh )] complexes. 3 3 3 3 spectrometer was equipped with a temperature cell regulator, HeLa (Human cervix adenocarcinoma) and RPE-1 (Human accuratewithin0.3°C.Solidsampleswereanalysedaspotass- retinalpigmentedepithelial)celllineswereusedtodetermine iumbromide(KBr)pelletsandthedatawerecollectedatroom the potential of our compounds. Cytotoxicity was assessed by temperature. fluorometric cell viability assay using resazurin. Worthy of All 1H and 13C NMR spectra were performed on a Bruker note, the highest concentration tested for fac-[Re(CO) (Trop) 3 300 MHz nuclear magnetic resonance spectrometer operating (PPh )] was 25 µM due to its poor solubility in cell media. 3 at 300 MHz using deuterated solvents. All 31P NMR spectra Cisplatin was used as a positive control. As can be seen in were performed on a Bruker 400 MHz nuclear magnetic reso- Table 4, complex fac-Re(CO) (Trop)(PPh ) (IC value 13.35 ± 3 3 50 nance spectrometer operating at 400 MHz using deuterated 0.94 µM) and fac-[Re(CO) (Flav)(PPh )] (IC value 12.21 ± 3 3 50 solvents. Chemical shifts are reported in parts per million 0.17 µM) were found to be the most cytotoxic compounds in (ppm)usingTMS(tetramethylsilane)asaninternalstandard. HeLa cell line with IC values comparable to the ones of cis- 50 platin. This could possibly relate to the higher stability of Synthesis these complexes as suggested by the kinetic studies. Interestingly, the IC 50 values of fac-Re(CO) 3 (Trop)(PPh 3 ) and Synthesis of fac-[NEt 4 ] 2 [Re(CO) 3 (Br) 3 ] (ReAA) (1). (NEt 4 )Br fac-[Re(CO) (Flav)(PPh )]onRPE-1cellswereallhigherthanon (5.06 g; 0.024 mol) was ground into a fine powder and dried 3 3 HeLa cells, showing an interesting selectivity between cancer- under vacuum overnight. Under dry nitrogen atmosphere ous and non-cancerous cells (IC 50 value >25 µM and 18.41 ± 2,5,8-trioxanone diglyme (150 ml) was added to the (NEt 4 )Br 3.16µM,respectively). andheatedto80°Cfor1hour.[Re(CO) 5 Br](5.02g;0.012mol) was added to the mixture and stirred for 24 hours at 120 °C. Aftercoolingdownthereactionwasfilteredandtheprecipitate Conclusion was washed with cold dichloromethane, followed by cold ethanol. The white product was left to dry overnight. (Yield: The kinetic studies add on to the growing body of work on 8.61g;90%)IR(KBr,cm −1):ν =1996,1846. co these complexes wherein it was found that complexes with Synthesis of fac-[NEt ] [Re(CO) (H O) ] (2). ReAA (100 mg; 42 3 2 3 bidentateligandslikeFlavandTrophavehighersecondorder 0.129 mmol) was dissolved in water (10 ml) at pH 2.2. AgNO 3 rate constants in general and that the mechanism of substi- (66.4 mg; 0.387 mmol) was added to the solution and it was tution is probably dissociative in nature. Furthermore, higher stirred at room temperature for 24 hours. AgBr (precipitate) stabilityconstantswerefoundfortheFlavandTropcomplexes wasfilteredoffandweighed(73.2mg).Bidentateligandswere coordinatedtoaphosphineligandinthesixthposition. addedtothefac-[NEt ] [Re(CO) (H O) ]filtrate. 42 3 2 3 Only fac-[Re(CO) (QuinH)(PTA)], fac-[Re(CO) (Trop)(PTA)], Synthesis of fac-[Re(CO) (Quin)(H O)] (3). 8-Hydroxy- 3 3 3 2 fac-[Re(CO) (Trop)(PPh )] and fac-[Re(CO) (Flav)(PPh )] com- quinoline (18.7 mg; 0.129 mmol) was added to the filtrate 3 3 3 3 plexes were used for biological studies due to solubility. and it was stirred at room temperature for 36 hours. The Compelling IC results were obtained for fac-[Re(CO) (Trop) solution was yellow and a bright yellow precipitate, fac- 50 3 (PPh )] and fac-[Re(CO) (Flav)(PPh )]. These compounds dis- [Re(CO) (Quin)(H O)],wasfilteredoff,driedandweighed.(Yield= 3 3 3 3 2 played the highest cytotoxicity of the series of compounds in 45.2mg;81%)IR(KBr,cm −1):ν =2021,1887.UV/Vis:λ = co max tested HeLa cells. Very interestingly, they had a smaller effect 265nm,ε=4000M −1cm −1.1HNMR(300MHz,methanol-d4): on non-cancerous cells, showing selectivity against malignant δ = 8.90 (dd, 1H, J = 1.2 Hz, 7.8 Hz), 8.63 (dd, 1H, J = 1.4 Hz, cellline.Thedatapoolistoosmalltoeffectivelyrelatekinetic 7.6 Hz), 7.85 (d, 1H, J = 7.4 Hz), 7.61 (d, 1H, J = 8.5 Hz), 7.58 datatocytotoxicityvalues,butitisinterestingtonotethatthe (d,1H,J=7.4Hz),7.12(d,1H,J=1.2Hz,8.4Hz),1.8(s,2H). FlavandTropphosphinecomplexesweremorestablethanthe 13C NMR (300 MHz, methanol-d4): δ = 161.2, 160.5, 159.3, Thisjournalis©TheRoyalSocietyofChemistry2020 DaltonTrans.,2020,49,35–46 | 41 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online Paper DaltonTransactions 143.2,135.1,128.2,125.3,122.6,120.4.Anal.calc.C,33.33,H, tate, fac-[Re(CO) (QuinH)(CH OH)], was dried and weighed. 3 3 1.86,N,3.24.Found:C,33.17,H,1.87N,3.26%. (Yield=24.4mg;78%)IR(KBr,cm −1):ν =2044,2008UV/Vis: co Synthesis of fac-[Re(CO) (diCl-Quin)(H O)] (4). 5,7-Dichloro- λ =367nm,ε=3650M −1cm −1.1HNMR(300MHz,metha- 3 2 max 8-hydroxyquinoline (27.6 mg; 0.129 mmol) was added to the nol-d4):δ=11.32(s,1H),9.87(s,1H),8.82(d,1H,J=4.6Hz), filtrate and it was stirred at room temperature for 36 hours. 7.82–7.85 (m, 3H), 3.62 (s, 1H), 3.45 (d, 2H, J = 6.2 Hz). 13C The solution was yellow and a light yellow precipitate, fac-[Re NMR (300 MHz, methanol-d4): δ = 175.3, 168.2, 147.8, 142.3, (CO) (diCl-Quin)(H O)], was filtered off, dried and weighed. 140.9, 135.7, 133.2, 128.1, 125.5, 116.7, 45.2. Anal. calc. C, 3 2 (Yield = 57.4 mg; 88%) IR (KBr, cm −1): ν = 2025, 1875. UV/ 34.68,H,2.13,N,2.70.Found:C,34.52,H,2.14,N,2.74%. co Vis: λ = 279 nm, ε = 4100 M −1 cm −1. 1H NMR (300 MHz, Synthesis of fac-[Re(CO) (Trop)(CH OH)] (9). fac-[Re max 3 3 methanol-d4):δ=9.72(dd,1H,J=1.7Hz.8.2Hz),9.65(d,1H, (CO) (Trop)(H O)] (50 mg; 0.122 mmol) was dissolved in 3 2 J=5.4 Hz),8.64(t,1H,J=3.2Hz,4.6 Hz),8.23(s,1H), 3.2(s, methanol (4 ml) and stirred at room temperature for 6 hours. 1H).13CNMR(300MHz,methanol-d4):δ=154.7,151.2,148.7, Thesolventwasevaporatedfromthesolution.Ayellowprecipi- 136.8,134.3,132.8,131.0,125.4,122.8.Anal.calc.C,28.75,H, tate, fac-[Re(CO) (Trop)(CH OH)], was dried and weighed. 3 3 1.21,N,2.79.Found:C,28.91,H,1.19,N,2.80%. (Yield = 49.6 mg, 94%) IR (KBr, cm −1): ν = 2034, 1877. UV/ co Synthesis of fac-[Re(CO) (QuinH)(H O)]·H O (5). Quinoline- Vis: λ = 345 nm, ε = 3100 M −1 cm −1. 1H NMR (300 MHz, 3 2 2 max 2,4-dicarboxylic acid (28.3 mg; 0.129 mmol) was added to the methanol-d4):δ=7.21(dt,2H,J=1.5Hz,8.2Hz),6.87(dt,2H, filtrate and it was stirred at room temperature for 36 hours. J=1.4Hz,5.5Hz),5.26(q,1H,J=8.2Hz),3.25(d,1H,J=5.2 The solution was orange and a bright orange precipitate, fac- Hz), 3.14 (d, 2H, J = 4.8 Hz). 13C NMR (300 MHz, methanol- [Re(CO) (QuinH)(H O)], was filtered off, dried and weighed. d4):δ=185.7,132.3,132.1,128.6,126.3,110.9,50.1.Anal.calc. 3 2 (Yield=60.7mg;92%)IR(KBr,cm −1):ν =2023,1994UV/Vis: C,31.20,H,2.14.Found:C,31.05,H,2.16%. co λ =362nm,ε=3675M −1cm −1.1HNMR(300MHz,metha- Synthesis of fac-[Re(CO) (Flav)(CH OH)] (10). fac-[Re max 3 3 nol-d4):δ=10.98(s,1H),8.54(s,1H),8.32(d,1H,J=5.4Hz), (CO) (Flav)(H O)](45mg;0.086mmol)wasdissolvedinmetha- 3 2 7.84–7.87(m,3H),2.21(s,2H).13CNMR(300MHz,methanol- nol (3 ml) and stirred at room temperature for 6 hours. The d4): δ = 174.5, 168.9, 145.2, 141.6, 133.4, 131.2, 130.9, 128.7, solventwasevaporatedfromthesolution.Ayellowprecipitate, 125.1, 117.3. Anal. calc. C, 32.19, H, 1.93, N, 2.68. Found: C, fac-[Re(CO) (Flav)(CH OH)], was dried and weighed. (Yield = 3 3 32.15,H,1.92,N,2.66%. 44.5mg;95%)IR(KBr,cm −1):ν =2003,1978.UV/Vis:λ = co max Synthesis of fac-[Re(CO) (Trop)(H O)] (6). Tropolone 325nm,ε=4255M −1cm −1.1HNMR(300MHz,methanol-d4): 3 2 (47.5 mg; 0.389 mmol) was added to the filtrate and it was δ=8.21(dd,2H,J=1.2Hz,4.4Hz),7.73–7.75(m,2H),7.71(t, stirred at room temperature for 36 hours. The solution was 1H, J = 7.4 Hz), 7.59 (d, 1H, J = 8.2 Hz), 7.48 (dd, 1H, J = 1.4 yellow and a bright yellow precipitate, fac-[Re(CO) (Trop) Hz, 9.2 Hz), 7.33 (d, 1H, J = 5.8 Hz), 7.05 (d, 1H, J = 6.2 Hz), 3 (H O)], was filtered off, dried and weighed. (Yield = 148 mg; 4.32(s, 1H), 3.44(s, 1H),3.18 (d, 2H,J=6.4 Hz),2.53(s, 1H). 2 93%)IR (KBr,cm −1):ν =2021, 1869. UV/Vis:λ = 343nm, 13C NMR (300 MHz, methanol-d4): δ = 158.2, 156.9, 152.3, co max ε=3120M −1cm −1.1HNMR(300MHz,methanol-d4):δ=6.44 143.2, 137.6, 135.1, 132.8, 129.8, 128.1, 127.6, 126.7, 122.5, (dt, 2H, J = 1.8 Hz, 8.5 Hz), 6.21 (dt, 2H, J = 1.4 Hz, 5.8 Hz), 121.8, 119.7, 45.7. Anal. calc. C, 42.30, H, 2.43. Found: C, 5.45 (q, 1H, J = 7.6 Hz), 3.87 (d, 1H, J = 4.8 Hz), 1.82 (s, 1H). 42.51,H,2.41%. 13C NMR (300 MHz, methanol-d4): δ = 188.3, 134.4, 134.2, Synthesis of fac-[Re(CO) (Quin)(PPh )] (11). fac-[Re 3 3 128.2, 127.8, 113.7, 82.8. Anal. calc. C, 29.34, H, 1.72. Found: (CO) (Quin)(H O)] (10 mg; 0.023 mmol) was dissolved in 3 2 C,29.17,H,1.71%. methanol (2 ml). PPh (6.32 mg; 0.0231 mmol) was dissolved 3 Synthesis of fac-[Re(CO) (Flav)(H O)] (7). 3-Hydroxyflavone in methanol (2 ml) separately and the two solutions were 3 2 (31.3 mg; 0.129 mmol) was added to the filtrate and it was added together. The mixture was stirred at room temperature stirred at room temperature for 36 hours. The solution was for 6 hours. The solution was left to crystalize. Brown crystals yellow and a light yellow precipitate, fac-[Re(CO) (Flav)(H O)], wereobtainedandsuitableforcollectionontheX-raydiffract- 3 2 was filtered off, dried and weighed. (Yield = 60.3 mg; 88%) IR ometer. (Yield = 13.5 mg; 88%) IR (KBr, cm −1): ν = 2015, co (KBr,cm −1):ν =1998,1943.UV/Vis:λ =320nm,ε=4280 1911, 1886. UV/Vis: λ = 435 nm, ε = 3560 M −1 cm −1. 31P co max max M −1cm −1.1HNMR(300MHz,methanol-d4):δ=8.33(dd,2H, NMR (400 MHz, toluene-d8): δ = 29 Hz. 1H NMR (300 MHz, J = 1.2 Hz, 4.4 Hz), 7.95–7.98 (m, 2H), 7.82 (t, 1H, J = 7.4 Hz), methanol-d4):δ=9.22(dd,1H,J=1.2,7.1Hz);8.56(d,1H,J= 7.63 (d, 1H, J = 8.2 Hz), 7.29 (dd, 1H, J = 1.4 Hz, 9.2 Hz), 7.21 4.2Hz),8.15(m,9H,PPh ),7.97(m,6H,PPh ),7.72(tt,1H,J= 3 3 (d,1H,J=5.8Hz),7.10(d,1H,J=6.2Hz),4.98(s,1H),3.21(s, 1.5,7.2Hz),7.64(dd,2H,J=1.7,6.4Hz).13CNMR(300MHz, 1H).13CNMR(300MHz,methanol-d4):δ=155.2,151.7,142.5, methanol-d4): δ = 165.4, 152.7, 138.2, 136.6, 135.8, 128.1, 139.8, 135.3, 134.2, 132.9, 131.5, 129.8, 127.3, 126.8, 124.4, 127.2, 126.8, 124.4, 121.7, 119.6. Anal. calc. C, 53.25, H, 3.13, 121.6, 118.1, 72.3. Anal. calc. C, 41.14, H, 2.11. Found: C, N,2.07,P,4.58.Found:C,52.99,H,3.15,N,2.09,P,4.55%. 40.90,H,2.12%. Synthesis of fac-[Re(CO) (diCl-Quin)(PPh )] (12). fac-[Re 3 3 Synthesis of fac-[Re(CO) (QuinH)(CH OH)] (8). fac-[Re (CO) (diCl-Quin)(H O)] (40 mg; 0.079 mmol) was dissolved in 3 3 3 2 (CO) (QuinH)(H O)] (30 mg; 0.059 mmol) was dissolved in methanol(2ml).PPh (21.2mg;0.079mmol)wasdissolvedin 3 2 3 methanol (3 ml) and stirred at room temperature for 6 hours. methanol (2 ml) separately and the two solutions were added Thesolventwasevaporatedfromthesolution.Ayellowprecipi- together. The mixture was stirred at room temperature for 42 | DaltonTrans.,2020,49,35–46 Thisjournalis©TheRoyalSocietyofChemistry2020 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online DaltonTransactions Paper 6hours.Thesolutionwaslefttocrystalize.Browncrystalswere (300 MHz, methanol-d4): δ = 184.7, 136.1, 135.8, 126.3, 124.7, obtainedandweresuitableforcollectionontheX-raydiffract- 118.6, 80.5, 33.2, 26.9, 19.8. Anal. calc. C, 49.99, H, 5.84, P, ometer. (Yield = 48.6 mg; 82%) IR (KBr, cm −1): ν = 2019, 4.60.Found:C,49.92,H,5.81,4.63%. co 1925, 1873. UV/Vis: λ = 453 nm, ε = 4125 M −1 cm −1. 31P Synthesis of fac-[Re(CO) (Flav)(PPh )] (16). fac-[Re(CO) (Flav) max 3 3 3 NMR (400 MHz, toluene-d8): δ = 32 Hz. 1H NMR (300 MHz, (H O)] (10 mg; 0.019 mmol) was dissolved in methanol (2 ml). 2 methanol-d4):δ=8.72(dd,1H,J=3.2,4.8Hz),8.54(d,1H,J= PPh (5.31 mg; 0.019 mmol) was dissolved in methanol (2 ml) 3 1.2 Hz), 7.92 (m, 9H, PPh ), 7.54 (s, 1H), 7.36 (m, 6H, PPh ). separately and the two solutions were added together. The 3 3 13C NMR (300 MHz, methanol-d4): δ = 154.7, 152.3, 149.8, mixturewasstirredat roomtemperature for6 hours.Thesolu- 140.7,135.6,133.2,132.9,128.6,124.1,118.7,116.3.Anal.calc. tion was left to crystalize. Yellow, thin, needle-like crystals, C,48.33,H,2.57,N,1.88,P,4.15.Found:C,48.24,H,2.54,N, not suitable for X-ray diffractometry were obtained. (Yield = 1.89,P,4.13%. 14.2 mg; 78%) IR (KBr, cm −1): ν = 1998, 1975, 1862. UV/Vis: co Synthesis of fac-[Re(CO) (Trop)(PPh )] (13). fac-[Re λ =422nm,ε=4425M −1cm −1.31PNMR(400MHz,toluene- 3 3 max (CO) (Trop)(H O)] (10 mg; 0.024 mmol) was dissolved in d8): δ = 23Hz.1H NMR(300MHz, methanol-d4): δ = 8.42(dd, 3 2 methanol(2ml).PPh (6.32mg;0.024mmol)wasdissolvedin 2H, J = 1.5 Hz, 5.4 Hz), 7.86 (m, 2H), 7.72 (t, 1H, J = 7.2 Hz), 3 methanol (2 ml) separately and the two solutions were added 7.68 (d, 1H, J = 7.2 Hz), 7.35 (m, 15H, PPh ), 7.16 (dd, 1H, J = 3 together. The mixture was stirred at room temperature for 1.8Hz,8.4Hz),7.13(d,1H,J=7.4Hz),7.06(d,1H,J=5.4Hz), 6 hours. The solution was left to crystalize. The crystals 4.76(s,1H).13CNMR(300MHz,methanol-d4):δ=157.4,153.8, obtained were not suitable forcollection on the X-ray diffract- 141.7, 137.6, 137.4, 136.1, 134.0, 132.8, 132.2, 131.8, 129.7, ometer. (Yield = 13.2 mg; 83%) IR (KBr, cm −1): ν = 2010, 128.3, 126.7, 126.4, 124.9, 122.1, 113.6, 70.6. Anal. calc. C, co 1934, 1887. UV/Vis: λ = 344 nm, ε = 3980 M −1 cm −1. 31P 56.17,H,3.14,P,4.02.Found:C,56.24,H,3.16,P,3.99%. max NMR (400 MHz, toluene-d8): δ = 27 Hz. 1H NMR (300 MHz, Synthesis of fac-[Re(CO) (Flav)(PTA)] (17). fac-[Re(CO) (Flav) 3 3 methanol-d4):δ=7.76(m,9H,PPh ),7.36(m,6H,PPh ),6.23 (H O)](20mg;0.038mmol)wasdissolvedinmethanol(2ml). 3 3 2 (dt, 2H, J = 1.4 Hz, 7.3 Hz), 5.84 (dt, 2H, J = 1.7 Hz, 5.4 Hz), PTA (5.97 mg; 0.038 mmol) was dissolved in methanol (2 ml) 5.31 (q, 1H, J = 5.4 Hz), 5.12 (d, 1H, J = 5.3 Hz). 13C NMR separately and the two solutions were added together. The (300 MHz, methanol-d4): δ = 184.7, 137.8, 136.5, 136.3, 135.4, mixture was stirred at room temperature for 6 hours. The 129.8,128.5,128.2,115.9,98.3.Anal.calc.C,51.45,H,3.08,P, product precipitated out and was recrystallized in methanol. 4.74.Found:C,50.97,H,3.11,P,4.76%. Thesolutionwaslefttocrystalize.Theyellowcrystalsobtained Synthesisoffac-[Re(CO) (Trop)(PTA)](14).fac-[Re(CO) (Trop) were not suitable for collection on the X-ray diffractometer. 3 3 (H O)](20mg;0.049mmol)wasdissolvedinmethanol(2ml). (Yield = 20.7 mg; 82%) IR (KBr, cm −1): ν = 2019, 1943. UV/ 2 co PTA (7.66 mg; 0.049 mmol) was dissolved in methanol (2 ml) Vis: λ = 415 nm, ε = 4532 M −1 cm −1. 31P NMR (400 MHz, max separately and the two solutions were added together. The toluene-d8):δ=−55Hz.1HNMR(300MHz,methanol-d4):δ= mixturewasstirredatroomtemperaturefor6hours.Thesolu- 8.26 (dd, 2H, J = 1.8 Hz, 6.2 Hz), 7.85 (m, 2H), 7.82 (t, 1H, J = tion was left to crystalize. The orange crystals obtained were 7.5 Hz), 7.73 (d, 1H, J = 5.2 Hz), 7.27 (dd, 1H, J = 1.5 Hz, 8.5 notsuitableforcollectionontheX-raydiffractometer.(Yield= Hz), 7.22 (d, 1H, J = 7.8 Hz), 7.18 (d, 1H, J = 5.4 Hz), 4.67 (s, 23.6mg;88%)IR(KBr,cm −1):ν =1982,1906.UV/Vis:λ = 1H), 4.21 (s, 6H, PTA), 2.66 (d, 6H, PTA, J = 5.3 Hz). 13C NMR co max 354 nm, ε = 4145 M −1 cm −1. 31P NMR (400 MHz, toluene-d8): (300 MHz, methanol-d4): δ = 157.2, 154.6, 147.3, 138.1, 134.8, δ=−60Hz.1HNMR(300MHz,methanol-d4):δ=6.11(dt,2H, 133.6, 132.3, 130.9, 128.5, 126.1, 125.7, 122.9, 120.6, 116.5, J=1.2Hz,6.4Hz),5.89(dt,2H,J=1.8Hz,7.4Hz),5.36(q,1H, 78.8, 89.3, 51.6. Anal. calc. C, 43.31, H, 3.33, N, 6.31, P, 4.65. J = 5.5 Hz), 4.28 (d, 1H, J = 1.8 Hz), 3.21 (s, 6H, PTA), 2.46 (d, Found:C,43.29,H,3.35,N,6.29,P,4.64%. 6H, PTA, J = 3.8 Hz). 13C NMR (300 MHz, methanol-d4): δ = Synthesisof fac-[Re(CO) (Flav)(PCy )] (18).fac-[Re(CO) (Flav) 3 3 3 186.3, 136.2, 133.1, 129.8, 125.9, 117.3, 93.4, 74.6, 52.8. Anal. (H O)](20mg;0.038mmol)wasdissolvedinmethanol(2ml). 2 calc. C, 34.97, H, 3.30, N, 7.65, P, 5.64. Found: C, 35.07, H, PCy (10.7 mg;0.038mmol)wasdissolvedinmethanol(2 ml) 3 3.28,N,7.60,P,5.62%. separately and the two solutions were added together. The Synthesis of fac-[Re(CO) (Trop)(PCy )] (15). fac-[Re mixture was stirred at room temperature for 6 hours. The 3 3 (CO) (Trop)(H O)] (30 mg; 0.073 mmol) was dissolved in yellow product precipitated out of solution and was filtered, 3 2 methanol(2ml).PCy (20.2mg;0.073mmol)wasdissolvedin driedandweighed.(Yield=30.2mg;71%)IR(KBr,cm −1):ν 3 co methanol (2 ml) separately and the two solutions were added = 2004, 1982. UV/Vis: λ = 456 nm, ε = 5530 M −1 cm −1. 31P max together. The mixture was stirred at room temperature for NMR (400 MHz, toluene-d8): δ = 57 Hz. 1H NMR (300 MHz, 6 hours. The solution was left to crystalize. The crystals methanol-d4): δ = 8.43 (dd, 2H, J = 2.1 Hz, 5.7 Hz), 7.98 (m, obtained were not suitable forcollection on the X-ray diffract- 2H), 7.84 (t, 1H, J = 6.5 Hz), 7.72 (d, 1H, J = 7.3 Hz), 7.48 (dd, ometer. (Yield = 40.3 mg; 82%) IR (KBr, cm −1): ν = 2011, 1H,J=1.8Hz,8.5Hz),7.26(d,1H,J=6.4Hz),7.18(d,1H,J= co 1964, 1856. UV/Vis: λ = 362 nm, ε = 5130 M −1 cm −1. 31P 6.8Hz),5.18(s,1H),1.67(m,18H,PCy ),1.52(m,12H,PCy ). max 3 3 NMR (400 MHz, toluene-d8): δ = 68 Hz. 1H NMR (300 MHz, 13C NMR (300 MHz, methanol-d4): δ = 154.8, 152.6, 141.2, methanol-d4):δ=6.78(dt,2H,J=2.4Hz,7.2Hz),6.43(dt,2H, 137.7, 134.6, 134.1, 131.3, 130.5, 128.5, 127.4, 126.1, 122.8, J=1.2Hz,6.8Hz),5.22(q,1H,J=6.8Hz),4.68(d,1H,J=1.5 121.9,116.5,78.5,32.4,29.6,27.1,22.7.Anal.calc.C,54.88,H, Hz), 1.66 (m, 18H, PCy ), 1.48 (m, 12H, PCy ). 13C NMR 5.37,P,3.93.Found:C,54.74,H,5.34,P,3.96%. 3 3 Thisjournalis©TheRoyalSocietyofChemistry2020 DaltonTrans.,2020,49,35–46 | 43 .MP 11:04:9 0202/2/1 no dedaolnwoD .9102 rebmevoN 11 no dehsilbuP View Article Online Paper DaltonTransactions Synthesis of fac-[Re(CO) (QuinH)(PTA)] (19). fac-[Re(CO) Julabo F-12 temperature cell regulator (accuratewithin 0.1 °C) 3 3 (QuinH)(H O)](20mg;0.039mmol)wasdissolvedinmethanol was used for the kineticmeasurements. All the kineticexperi- 2 (2 ml). PTA (6.23 mg; 0.039 mmol) was dissolved in methanol mentsweredoneunderpseudofirst-orderconditions,withthe (2 ml) separately and the two solutions were added together. ligandinexcess.ScientistMicomath,Version2.01983wasused The mixture was stirred at room temperature for 6 hours. The tofitthedatatoselectedfunctions.Thecomputerleastsquare solution was left to crystalize. The orange product precipitated fits of data is represented by the solid lines in the figures, out of solution and was filtered, dried and weighed. (Yield = while the individual points representthe experimental values, 23.2 mg; 89%) IR (KBr, cm −1): ν = 2046, 1974. UV/Vis: λ = denotedbyselectedsymbols. co max 442nm,ε=3870M −1cm −1.31PNMR(400MHz,toluene-d8):δ = −67 Hz. 1H NMR (300MHz, methanol-d4): δ = 10.12 (s, 1H), Celllinesandcellculture 8.63 (s, 1H), 8.55 (d, 1H, J = 7.3 Hz), 7.72 (m, 3H), 3.27 (s, 6H TheHeLa(Homosapienscervixadenocarcinoma)celllinewas (PTA)),2.58(d,6H,PTA,J=5.6Hz).13CNMR(300MHz,metha- cultured in DMEM (Gibco, Life Technologies, USA) sup- nol-d4):δ=178.6,169.4,147.1,145.5,131.2,130.6,129.8,125.3, plementedwith10%offetalcalfserum(Gibco).RPE-1(Homo 123.8,114.9,86.4,41.2.Anal.calc.C,37.33,H,2.80,N,8.71,P, sapiens retinal pigmented epithelial) cell line was cultured in 4.81.Found:C,37.28,H,2.81,N,8.68,P,4.79%. DMEM/F-12 (Gibco) supplemented with 10% of fetal calf Synthesis of fac-[Re(CO) (Trop)(PPh ) ]·2C H (20). fac- serum. Both cell lines were complemented with 100 U ml −1 2 32 7 8 [Re(CO) (Trop)(PPh )] (10 mg; 0.015 mmol) was dissolved in penicillin–streptomycin mixture (Gibco), and maintained in 3 3 toluene (2 ml). An excess of PPh (24.6 mg; 0.092 mmol) was humidifiedatmosphereat37°Cand5%ofCO . 3 2 dissolved in toluene (2 ml) separately and the two solutions Cytotoxicity were added together. The mixture was heated to 80 °C, whilst stirred for 24 hours. The solvent was evaporated, orange crys- Cytotoxicity was assessed by fluorometric cell viability assay tals, suitable for X-ray diffractometry were obtained. (Yield = using resazurin (ACROS Organics). HeLa and RPE-1 cells were 8.38mg;72%)IR(KBr,cm −1):ν =1919,1832.UV/Vis:λ = seededintriplicatesin96wellplatesatadensityof4000cells co max 312 nm, ε = 6945 M −1 cm −1. 31P NMR (400 MHz, toluene-d8): per well in 100 µl, 24 h prior to treatment. Cells were then δ = 26, −7 Hz. 1H NMR (300 MHz, methanol-d4): δ = 7.79 (m, treated with increasing concentration of compounds for 48 h. 12H, PPh ), 7.38 (m, 12H, PPh ), 7.10 (m, 10H, toluene), 6.45 After that time medium was replaced by fresh complete 3 3 (dt, 2H, J = 1.6 Hz, 7.5 Hz), 5.63 (dt, 2H, J = 1.5 Hz, 5.5 Hz), medium containing resazurin (0.2 mg ml −1 final concen- 5.22 (q, 1H, J = 5.2 Hz), 5.08 (d, 1H, J = 5.4 Hz), 2.44 (s, 6H, tration). After 4 h incubation at 37 °C, fluorescence signal of toluene). 13C NMR (300 MHz, methanol-d4): δ = 185.3, 138.2, resorufin product was read by SpectraMax M5 microplate 137.3, 137.1, 134.5, 129.6, 128.9, 128.7, 128.2, 127.5, 124.6, reader (ex: 540 nm em: 590 nm). IC values were calculated 50 117.8, 95.2, 25.4. Anal. calc. C, 57.09, H, 4.14, P, 4.99. Found: usingGraphPadPrismsoftware. C,57.02,H,4.18,P,5.02%. X-raydatacollectionandrefinement Conflicts of interest The reflection data used to determine the structures reported here were collected on a Bruker X8 Apex II 4K diffractometer, Therearenoconflictstodeclare. using graphite monochromated Mo-Kα radiation (with wave- lengthλ=0.71073Å)andwithω-andφ-scansatatemperature Acknowledgements of100K. SAINT-Plus77 wasusedto doall thecellrefinements and the data reduction was done with SAINT-Plus and The authors thank the University of the Free State Research XPREP.77 The multi-scan technique and software package Fund (H. G. V.), the South African NRF (H. G. V.) and SADABS78 was used to correct the absorption effects. The the ERC (Consolidator Grant PhotoMedMet GA 681679 crystal structure was solved using the SIR-9779 package and to G. G.). This work has received support under the refined with WinGX80 and SHELXL-97.81 The graphical rep- program “Investissements d’ Avenir” launched by the French resentations of the crystal structures were obtained using the Government and implemented by the ANR with the reference program DIAMOND.82 Unless otherwise stated, all the struc- ANR-10-IDEX-0001-02PSL(G.G.). tures are shown with thermal ellipsoids drawn at 50% prob- ability level. All non-hydrogen atoms are refined anisotropi- cally. Aromatic and hydroxyl hydrogen atoms were placed in References geometricallyidealizedpositions(C–H=0.93andO–H=0.85) and constrained to ride on their parent atoms (U (H) = 1 E. B. Bauer, A. A. Haase, R. M. Reich, D. C. Crans and iso 1.5U (C)and1.5U (O)). F.E.Kühn,Coord.Chem.Rev.,2019,393,79. eq eq 2 A.A.Haase,E.B.Bauer,F.E.KühnandD.C.Crans,Coord. Kineticexperiments Chem.Rev.,2019,394,135. All reagents and chemicals were of analytical grade. 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