a b c Supplementary Figure S1 a, wireframe view of the crystal structure of compound 11. b, view of the pyridinium sites. c, crystal packing of compound 11. 1
a b c Supplementary Figure S2 a, wireframe view of the crystal structure of compound 12. b, schematic view of the position of the pyridinium cations. c, crystal packing showing the fixed parts of the structure corresponding to the octahedral core, green, and the mobile parts corresponding to the octyl chains and pyridinium, red. 2
a b c d Supplementary Figure S3 1 H-NMR spectra in DMF d 7. a, cage 1 (purple), b, calix[4]arene 7 and pyridine (4 equiv, green), c, compound 7 and uranyl nitrate (1.33 equiv, red), d, compound 7 alone (blue). In all cases compound 7 was at 6 mm concentration. 3
a b c 4
d Supplementary Figure S4 DOSY-NMR spectra in DMF d 7 at 300 K. a, compound 7, b, compound 7 and uranyl nitrate (1.33 equiv), c, calix[4]arene 7 and pyridine (4 equiv), d, cage 1. In all cases compound 7 was at 6 mm concentration. 5
a b c d a b c d Supplementary Figure S5 1 H-NMR full spectra (top) and zoom (bottom) in DMF d 7. a, cage 2 (purple), b, calix[5]arene 8 and pyridine (5 equiv, green), c, compound 8 and uranyl nitrate (1.7 equiv, red), d, compound 8 alone (blue). In all cases calix[5]arene 8 was at 12 mm concentration. 6
a b c 7
d Supplementary Figure S6 DOSY-NMR spectra in DMF d 7 at 300 K. a, compound 8, b, calix[5]arene 8 and uranyl nitrate (1.7 equiv), c, calix[5]arene 8 and pyridine (5 equiv) and d, cage 2. In all cases compound 8 was at 12mM concentration. 8
Supplementary Figure S7 1 H-NMR spectrum of cage 11 in DMF d 7. Component 9 is at 16 mm concentration. Signals at 7.46 and 1.20 ppm correspond to ligand 9, whereas signals at 8.35 and 2.22 ppm correspond to cage 11. 9
Supplementary Figure S8 DOSY NMR spectrum of cage 11 in DMF d 7 at 300 K. Ligand 9 was at 6 mm concentration. 10
Supplementary Figure S9 1 H-NMR spectrum of cage 12 in 1:1 DMF d 7 :TCE d 2. Ligand 10 is at 19 mm concentration. Signals at 7.38 and 1.95 ppm correspond to ligand 10, whereas signals at 8.25 and 2.22 ppm correspond to cage 12. 11
Supplementary Figure S10 DOSY NMR experiment of cage 12 in 1:1 DMF d 7 :TCE d 2 at 300 K. Compound 10 was at 19 mm concentration. 12
Supplementary Figure S11 DLS experiments in DMF at 293 K. Composition of DLS spectra of cage 1 (red line), 2 (green line), 11 (blue line) and 12 (black line). 13
Supplementary Figure S12. Ortep drawing of cage 1. (Ellipsoids are drawn at 50% probability. Carbon: black, Oxygen: red, Uranium: green). 14
Supplementary Figure S13. Ortep drawing of cage 1 with cyclen. (Ellipsoids are drawn at 50% probability. Carbon: black, Nitrogen: blue, Oxygen: red, Uranium: green). 15
Supplementary Figure S14. Ortep drawing of cage 2. (Ellipsoids are drawn at 50% probability. Carbon: black, Oxygen: red, Uranium: green). 16
Supplementary Figure S15. Ortep drawing of cage 11. (Ellipsoids are drawn at 50% probability. Carbon: black, Oxygen: red, Uranium: green). 17
Supplementary Figure S16. Ortep drawing of cage 12. (Ellipsoids are drawn at 50% probability. Carbon: black, Oxygen: red, Uranium: green). 18
a b c Supplementary Figure S17. NMR and HRMS spectra of compound 6: a, 1 H-NMR spectrum in d 6 -DMSO, b, 13 C- NMR spectrum in d 6 -DMSO, c, ESI-HRMS. 19
a b c Supplementary Figure S18. NMR and HRMS spectra of compound 8: a, 1 H-NMR spectrum in d 6 -DMSO, b, 13 C- NMR spectrum in d 6 -DMSO, c, MALDI-HRMS. 20
Cage [ligand] [UO 2+ 2 ] (mm) (mm) I cage I ligand K app (mm -13 ) 1 19.0 25.2 1.00 0.63 1.82 x 10-10 11 19.0 25.2 1.00 1.69 6.05 x 10-16 12 19.0 25.2 1.00 2.89 4.72 x 10-18 Supplementary Table S1. Apparent constants of formation of cages 1, 11 and 12 calculated from the equation reported above. 1 H-NMR spectra were measured in DMF d 7 at 300 K. Cage [ligand] mm [UO 2 2+ ] mm I cage I ligand K app (mm -31 ) 2 12.0 20.0 1.00 1.56 1.75 x 10-31 Supplementary Table S2. Stability constants for cage 2 calculated from the equation reported above. 1 H-NMR spectrum was measured in DMF d 7 at 300 K. 21
Supplementary methods Procedures 25,26,27,28-Tetrahydroxycalix[4]arene-5,11,17,23-tetracarboxylic acid (7) To a stirred mixture of tetraformylcalix[4]arene (5) (381 mg, 0.710 mmol) in DMSO (7.1 ml) and sodium dihydrogen phosphate (85 mg, 0.710 mmol) in 1 ml of water, a solution of sodium chlorite (642 mg, 5.68 mmol) in 6 ml of water was added drop-wise over 3h at room temperature. After stirring overnight at room temperature, the reaction mixture was acidified with 5M aq. HCl and the solid formed was filtered, washed with water and dried to obtain compound 7 as a light yellow solid (368 mg, 86%). 1 H-NMR (400 MHz, dimethylsulphoxide-d 6 (DMSO-d 6 ), 398K) δ 7.64 (s, 8H, CH), 3.91 (s, 8H, CH 2 ). 13 C- NMR (100 MHz, DMSO-d 6 ) δ 167.5 (C quat -OH), 158.1 (COOH), 130.6 (CH), 129.5 (C- CH 2 ), 121.6 (C-COOH). HRMS (m/z): [M - ] calcd. for C 32 H 23 O 12 599.1195; found 599.1220. Characterisation of cages in the solid state Single crystal X-ray diffraction data of 1. The measured crystal was removed from the mixture, placed on a slide and quickly covered with vacuum grease and Paratone-N as protecting oil. Afterwards, it was scooped with a Cryoloop and mounted on the goniometer head. Single crystal X-ray diffraction data of 1 were collected on a Bruker Kappa APEX II DUO diffractometer equipped with an APPEX 2 4K CCD area detector, a Microsource with Cu Kα radiation and an Oxford Cryostream 700 low temperature device (T = -173 C). Full-sphere data collection was used with ω and ϕ scans. Programs used: Data collection Apex2 V2009 1.0 (Bruker- Nonius 2008), data reduction Saint + Version 7.60A (Bruker AXS 2008) and absorption correction SADABS V. 2008-1 (2008). The structure solution and refinement were carried out using SHELXTL V6.14 46 22
Crystallographic data at 100(2) K: C 96 H 60 O 59 U 4, Mr = 3109.56 gmol -1, Crystal dimensions: 0.20 x 0.15 x 0.15 mm 3, cubic, space group: Fm-3m, a = b = c = 33.702(2) Å, V = 38281(4) Å 3, Z = 8, ρ calc = 1.079 Mg/m 3, R 1 = 0.1166 (0.1207), wr 2 = 0.3713 (0.3773), for 621 reflections with I>2σ(I) (for 683 reflections [R int : 0.0718] ) with a total of 39889 reflections measured, 2θ=81.84º, 77 parameters and 130 restrains. CCDC deposit number 856283. Residual electron densities in the solvent-accessible void, due to disordered solvent molecules as well as counterions, were treated with the PLATON SQUEEZE program 47 Before this treatment the maximum remained electron density was 6.37, R 1 = 0.2384 and wr 2 = 0.5797 for all data with 529 unique reflections out of 683 with I > 2σ(I). The asymmetric unit is made up of 1/8 of a calix[4]arene molecule, 1/6 of a uranyl cation unit and three water molecules with occupancy of ¼, ¼ and 1/8. Two of them lie on a 48i Wyckoff site and one in the 24e becoming a full water molecule in the unit cell. Anisotropic displacements of the calix[4]arene were restrained and water molecules were also restrained so they approximated to isotropic behaviour. Hydrogen atom positions of all water molecules were neither located nor calculated. Single crystal X-ray diffraction data of 1 with cyclen. The measured crystal was removed from the mixture, placed on a slide and quickly covered with vacuum grease and Paratone-N as protecting oil. Afterwards, it was scooped with a Cryoloop and mounted on the goniometer head. Single crystal X-ray diffraction data were collected on a Bruker Kappa APEX II DUO diffractometer equipped with an APPEX 2 4K CCD area detector, a Microsource with Mo Kα radiation and an Oxford Cryostream 700 low temperature device (T = -173 C). Full-sphere data collection was used with ω and ϕ scans. Programs used: Data collection, Apex2 V2009 1.0 (Bruker- Nonius 2008), data reduction, Saint + Version 7.60A (Bruker AXS 2008) and absorption correction, SADABS V. 2008-1 (2008). Structure solution and refinement were carried out using SHELXTL V6.14 46 Crystallographic data at 100(2) K: C 208 H 168 N 8 O 96 U 8, Mr = 6219.74 gmol -1, Crystal dimensions: 0.10 x 0.05 x 0.05 mm 3, tetragonal, space group: I4/m, a = b = 23.477(3) Å, c = 30.143(3) Å, V = 16614(3) Å 3, Z = 2, ρ calc = 1.243 Mg/m 3, R 1 = 0.0725 (0.1187), wr 2 = 0.2045 (0.2186), for 3943 reflections with I>2σ(I) (for 6864 reflections [R int = 0.0709] ) 23
with a total of 50613 reflections measured, 2θ=48.58º, 538 parameters and 1097 restrains. CCDC deposit number 856284. Residual electron densities in the solvent-accessible void due to disordered solvent molecules were treated with the PLATON SQUEEZE program 47 Before the treatment of SQUEEZE program the maximum remained electron density was 2.11, R 1 = 0.1736 and wr 2 = 0.3144 for all data with 3844 unique reflections out of 6864 with I > 2σ(I). The asymmetric unit is made up of ¾ of a calix[4]arene molecule, one uranyl cation unit, ¼ of cyclen molecule and one water molecule. The ¾ of the calix[4]arene molecule was distributed in two calix[4]arene molecules in which ¼ and ½ are present in the asymmetric unit. The uranyl cation unit is disordered over two positions with 50% occupancy in each site and the anisotropic displacement parameters were restrained. As a result, the ¼ of the calix[4]arene molecule was disordered over two positions (50:50) and one of the carboxylates groups of the other molecule is also disordered over two positions (50:50). Anisotropic displacements of the calix[4]arene were restrained. The cyclen molecule is distributed over two positions in which only 1/8 of the molecule is present. Distances of the nitrogen and carbon atoms were restrained as well as anisotropic displacement parameters. Two of the water molecules lie in a 4e and 8h Wyckoff sites and the remaining water molecule occupancy has been fixed to 25% of occupancy. Hydrogen atom positions of all water molecules were neither located nor calculated. Single crystal X-ray diffraction data of 2. The measured crystal was removed from the mixture, placed on a slide and quickly covered with vacuum grease and Paratone-N as protecting oil. Afterwards, it was scooped with a Cryoloop and mounted on the goniometer head. Single crystal X-ray diffraction data were collected on a Bruker Kappa APEX II DUO diffractometer equipped with an APPEX 2 4K CCD area detector, a Microsource with Mo Kα radiation and an Oxford Cryostream 700 low temperature device (T = -173 C). Full-sphere data collection was used with ω and ϕ scans. Programs used: Data collection Apex2 V2009 1.0 (Bruker- Nonius 2008), data reduction Saint + Version 7.60A (Bruker AXS 2008) and absorption correction SADABS V. 2008-1 (2008). Structure solution and refinement were carried out using SIR2008 48 and SHELXTL V6.14 46, respectively. 24
Crystallographic data at 100(2) K: C 490 H 312 N 1 O 232 U 20, Mr = 14700.03 gmol -1, crystal dimensions: 0.10 x 0.10 x 0.10 mm 3, orthorhombic, space group: Cmca, a = 54.142(4) Å, b = 50.412(4) Å, c = 42.872(4) Å, V = 117017(17) Å 3, Z = 4, ρ calc = 0.834 Mg/m 3, R 1 = 0.1479 (0.2319), wr 2 = 0.4029 (0.4379), for 11064 reflections with I>2σ(I) (for 19565 reflections [R int : 0.1053] ) with a total of 168811 reflections measured, 2θ=35.5º, 1726 parameters and 1614 restrains. CCDC deposit number 856285. The asymmetric unit contains three calix[5]arene molecules and five uranyl units over six positions, two of them lying on a 8f Wickoff site. Half pyridinium molecule was located as well as five water positions. The distances within the pyridinium molecule were constrained. Anisotropic displacements of the calix[5]arene molecules were restrained and water and pyridinium molecules were restrained so they approximate to isotropic behaviour. Hydrogen atom positions of all water molecules were neither located nor calculated. Accessible void remains in the structure since a jumble of residual peak could not be modelled with chemical sense. Single crystal X-ray diffraction data of 11. The measured crystal was removed from the solution, placed on a slide and quickly covered with vacuum grease and Paratone-N as protecting oil. Afterwards, it was scooped with a Cryoloop and mounted on the goniometer head. Single crystal X-ray diffraction data of 11 were collected on a Bruker-Nonius diffractometer equipped with an APPEX 2 4K CCD area detector, a FR591 rotating anode with Mo Kα radiation, Montel mirrors and an Oxford Cryostream 700 low temperature device (T = -173 C). Full-sphere data collection was used with ω and ϕ scans. Programs used: Data collection Apex2 V2009 1.0 (Bruker-Nonius 2008), data reduction Saint + Version 7.60A (Bruker AXS 2008) and absorption correction SADABS V. 2008-1 (2008). Structure solution and Refinement were carried out using SIR2008 48 and SHELXTL V6.14 46, respectively. Crystallographic data at 100(2) K: C 278 H 279 N 3 O 88 U 8, Mr = 6974.28gmol -1, Crystal dimensions: 0.10 x 0.02 x 0.02 mm 3, orthorhombic, space group: Ibam, a = 24.8284(13) Å, b = 40.177(2) Å, c = 41.4124(19) Å, V = 41310(4) Å 3, Z = 4, ρ calc = 1.121 Mg/m 3, R 1 = 0.1564 (0.2838), wr 2 = 0.3850 (0.4272), for 7947 reflections with I>2σ(I) (for 16698 reflections [R int : 0.1530] ) with a total of 97093 reflections measured, 2θ=48.74º, 839 25
parameters and 1489 restrains. CCDC deposit number 856286. Residual electron densities in the solvent-accessible void due to disordered solvent molecules as well as counterions were treated with the PLATON SQUEEZE program 47. Before the treatment of SQUEEZE program the remaining maximum electron density was 18.37, R 1 = 0.2859 and wr 2 = 0.4276 for all data with 7947 unique reflections out of 16698 with I > 2σ(I). The asymmetric unit is made up in total of one and a half O-propyl calix[4]arene molecule, two uranyl units, and one pyridinium over two positions. The O-propyl chains which showed disorder were modelled in two positions with 50% of occupancy in each site. Distances within O-propyl chains and pyridinium molecules were restrained as well as the anisotropic displacements of the O-propyl chains, the calix[4]arenes and pyridinium molecules. 26
Single crystal X-ray diffraction data of 12 The measured crystal was removed from the mixture, placed on a slide and quickly covered with vacuum grease and Paratone-N as protecting oil. Afterwards, it was scooped with a Cryoloop and mounted on the goniometer head. Single crystal X-ray diffraction data were collected on a Bruker Kappa APEX II DUO diffractometer equipped with an APPEX 2 4K CCD area detector, a Microsource with Mo Kα radiation and an Oxford Cryostream 700 low temperature device (T = -173 C). Full-sphere data collection was used with ω and ϕ scans. Programs used: Data collection Apex2 V2009 1.0 (Bruker- Nonius 2008), data reduction, Saint + Version 7.60A (Bruker AXS 2008) and absorption correction, SADABS V. 2008-1 (2008). Structure solution and Refinement were carried out using SHELXTL V6.14 46. Crystallographic data at 100(2) K: C 424 H 544 N 8 O 94 U 8, Mr = 9160.91 gmol -1, Crystal dimensions: 0.20 x 0.10 x 0.10 mm 3, tetragonal, space group: I4 1 /a, a = b = 34.870(3) Å, c = 38.318(4) Å, V = 46592(8) Å 3, Z = 4, ρ calc = 1.306 Mg/m 3, R 1 = 0.1541 (0.2791), wr 2 = 0. 0.3850 (0.4319), for 6630 reflections with I>2σ(I) (for 13608 reflections [R int : 0.0945]) with a total of 80486 reflections measured, 2θ=43.32º, 1212 parameters and 1638 restrains. CCDC deposit number 856287. The capsule in asymmetric unit is made up of one and a half O-octyl calix[4]arene molecules and two uranyl units. One of the uranyl units was disordered over two positions with a 50% of occupancy in each site. Consequently, the two carboxylate groups coordinated to the uranyl cation were also disordered (50:50). Distances of the oxygen and carbon atoms in the O-octyl chains were restrained. Moreover, distances between atoms of the neighbouring O-octyl were been restrained to fulfil the minimum required distances between neighbouring atoms. Anisotropic displacements of the O-octyl chains and the calix[4]arenes were restrained. The charges were balanced by two pyridinium molecules, one of them disordered over two sites (75:25). Anisotropic displacements were restrained. Two water molecules were also present in the asymmetric unit, one of them lying in a 8e Wyckoff site and the hydrogen atom positions were neither located nor calculated. 27
Characterisation of cages in solution DOSY NMR diffusion coefficients, calculated on the basis of monodimensional fitting and expressed in cm 2 /s are: 7 (3.50 E-06), DMF-d 7 ((1.45 ± 0.07) E-05) 7 and uranyl nitrate (1.33 eq) (3.40 E-06), DMF-d 7 ((1.44 ± 0.07) E-05) 7 and pyridine (4 eq) (3.51 E-06), DMF-d 7 ((1.45 ± 0.05) E-05) Cage 1 ((1.76 ± 0.05) E-06), free ligand 7 (3.26 E-06), DMF-d 7 ((1.4 ± 0.1) E-05) DOSY NMR diffusion coefficients, calculated on the basis of monodimensional fitting and expressed in cm 2 /s are: 8 ((3.02 ± 0.01) E-06), DMF-d 7 ((1.46 ± 0.01) E-05) 8 and uranyl nitrate (1.7 eq) ((2.75 ± 0.02) E-06), DMF-d 7 ((1.36 ± 0.02) E-05) 8 and pyridine (5 eq) ((2.99 ± 0.01) E-06), DMF-d 7 ((1.46 ± 0.03) E-05) Cage 2 ((1.5 ± 0.3) E-06), free ligand 8 ((2.3 ± 0.2) E-06), DMF-d 7 ((1.3 ± 0.2) E- 05). DOSY NMR diffusion coefficients, calculated on the basis of monodimensional fitting and expressed in cm 2 /s are: cage 11 ((1.76 ± 0.09) E-06), 9 ((3.09 ± 0.07) E-06), DMF-d 7 ((1.4 ± 0.1) E-05). DOSY NMR diffusion coefficients, calculated on the basis of monodimensional fitting and expressed in cm 2 /s, are: Cage 12 ((6.7 ± 0.2) E-07), 10 ((1.259 ± 0.009) E-06), DMF d 7 ((6.68 ± 0.02) E-06), TCE d 2 (5.46 E-06). 28
Dynamic Light Scattering (DLS) Solutions in DMF of mixture of components forming the cages were analyzed by a Dynamic Light Scattering instrument. Corrections were carried out only for the solvent (refractive index and viscosity). Calix[4]arene 7 (17 mm concentration), uranyl nitrate (1.33 equiv) and pyridine (4 equiv) Calix[5]arene 8 (10 mm concentration), uranyl nitrate (1.7 equiv) and pyridine (5 equiv) Calix[4]arene 9 (13 mm concentration), uranyl nitrate (1.33 equiv) and pyridine (4 equiv) Calix[4]arene 10 (13 mm concentration), uranyl nitrate (1.33 equiv) and pyridine (4 equiv) Average hydrodynamic radii observed: 1: 1.226 ± 0.002 nm 2: 1.699 ± 0.001 nm 11: 1.236 ± 0.005 nm 12: 1.472 ± 0.016 nm. Stability constants The equilibrium for the octahedral cages can be expressed by the following equation: 6 7 + 8 UO 2 (NO 3 ) 6H 2 O + 24Py Cage 1 + 16 Py-H + + 16 NO 3 - + 48 H 2 O Assuming that the deprotonation of calixarene 7 is quantitative and the uranyl nitrate is completely dissociated, we can neglect in the equilibrium the species Py, Py-H +, NO 3 -. 29
The apparent overall formation constants of octahedral cages 1, 11 and 12 can be calculated as follows from the relative areas of the aromatic CH peaks (ligand and cage) in the corresponding NMR spectra (equation illustrated for cage 1): K app = [Cage 1]/ {[7] 6 [UO 2+ 2 ] 8 eq } [Cage 1] and [7] at equilibrium can be calculated by integration of the corresponding proton signals in the 1 H-NMR spectrum. On the other hand, [UO 2+ 2 ] eq can be obtained by subtracting eight equivalents of cage from the initial concentration of uranyl nitrate [UO 2+ 2 ]. K app = [Cage 1]/ {[7] 6 ([UO 2+ 2 ] -8 [Cage 1]) 8 } The equilibrium for the icosahedral cages can be expressed by the following equation: 12 8 + 20 UO 2 (NO 3 ) 2 6H 2 O + 60Py Cage 2 + 40 Py-H + + 40NO 3 - + 120H 2 O The apparent overall constant of formation of the icosahedral cage 2 can be calculated by the following equation, after assuming the same approximations done in the previous case: K app = [Cage 2] / {[8] 12 x [UO 2 ] 20 eq } [Cage 2] and [8] at the equilibrium can be calculated by integration of the corresponding proton signals in the 1 H-NMR. Instead, [UO 2+ 2 ] eq can be obtained by subtracting twenty equivalents of cage from the initial concentration of uranyl nitrate [UO 2+ 2 ]. K app = [Cage 2] / {[8] 12 x ([UO 2+ 2 ] 20 x [Cage 2]) 20 } 30
Supplementary references 46 Sheldrix, G. M. SHELXTL Crystallographic System Ver 6.14, Bruker AXS Inc., Madison, Wisconsin, 2000. 47 Spek, A. L. Acta Crystallogr., Sect. A: Fundam. Crystallogr., 46, C34 (1990). 48 Caliandro, R. et al. Advances in ab initio protein phasing by Patterson deconvolution techniques. J. Appl. Cryst. 40, 883-890 (2008). 31