Rhodium(III) Dihalido Complexes: The Effect of Ligand Substitution and Halido Coordination on Increasing Cancer Cell Potency.

eprints@whiterose.ac.uk https://eprints.whiterose.ac.uk Universities of Leeds, Sheffield and York Deposited via The University of Leeds. White Rose Research Online URL for this paper: https://eprints.whiterose.ac.uk/id/eprint/171119/ Version: Supplemental Material Article: Lord, RM, Zegke, M, Basri, AM et al. (2021) Rhodium(III) Dihalido Complexes: The Effect of Ligand Substitution and Halido Coordination on Increasing Cancer Cell Potency. Inorganic Chemistry, 60 (3). pp. 2076-2086. ISSN: 0020-1669 https://doi.org/10.1021/acs.inorgchem.0c03704 © 2021 American Chemical Society. This is an author produced version of an article, published in Inorganic Chemistry. Uploaded in accordance with the publisher's selfarchiving policy. Reuse Items deposited in White Rose Research Online are protected by copyright, with all rights reserved unless indicated otherwise. They may be downloaded and/or printed for private study, or other acts as permitted by national copyright laws. The publisher or other rights holders may allow further reproduction and re-use of the full text version. This is indicated by the licence information on the White Rose Research Online record for the item. Takedown If you consider content in White Rose Research Online to be in breach of UK law, please notify us by emailing eprints@whiterose.ac.uk including the URL of the record and the reason for the withdrawal request. Electronic Supplementary Information Rhodium(III) dihalido complexes: the effect of ligand substitution and halido coordination on increasing cancer cell potency Rianne M. Lord,[a,b]* Markus Zegke,[b] Aida M. Basri,[c] Christopher M. Pask[c] and Patrick C. McGowan[c] a School of Chemistry, University of East Anglia, Norwich Research Park, Norwich, NR4 7TJ, U.K. Email: r.lord@uea.ac.uk b School of Chemistry and Biosciences, University of Bradford, Bradford, BD7 1DP, U.K. c School of Chemistry, University of Leeds, Woodhouse Lane, Leeds, LS2 9JT, U.K. 1 Electronic Supplementary Information X-ray Crystallographic Analysis Table S 1 X-ray crystallographic data for complexes 1, 3 ( and ), 4a and 4b, s.u.s shown in parenthesis Complex 1 3 () 3 () 4a 4b CCDC Number 1978080 1978078 2042715 1978082 1978081 Empirical formula C25H21Cl2F2N4O3Rh C24H17Br2Cl2N4O2Rh C24H17Br2Cl2N4O2Rh C26H25Cl2I2N4O4Rh C27H24Cl2I2N5O3Rh Formula weight 637.278 727.046 727.046 885.132 894.143 Temperature/K 100.1(4) 99.9(3) 100.3(9) 100.2(4) 120.3(7) Crystal system orthorhombic monoclinic monoclinic monoclinic monoclinic Space group P212121 Cc Cc P21/c P21/n a/Å 10.5456(3) 9.3613(3) 9.3527(2) 17.9801(8) 8.1441(3) b/Å 12.0060(4) 23.8677(7) 23.9723(6) 10.3200(4) 32.8058(12) c/Å 19.5775(7) 11.4997(3) 11.5084(3) 17.7408(9) 11.0381(4) α/° 90 90 90 90 90 β/° 90 100.495(3) 100.627(2) 115.036(6) 90.522(3) γ/° 90 90 90 90 90 Volume/Å3 2478.72(14) 2526.43(13) 2535.99(11) 2982.6(3) 2948.98(18) Z 4 4 4 4 4 ρcalcg/cm3 1.708 1.911 1.904 1.971 2.014 μ/mm-1 0.956 4.081 4.065 2.860 23.116 F(000) 1277.3 1411.6 1411.6 1698.2 1729.4 Crystal size/mm3 0.478 × 0.202 × 0.132 0.093 × 0.06 × 0.04 0.21 x 0.15 x 0.09 0.412 × 0.366 × 0.316 0.102 × 0.072 × 0.035 Radiation Mo Kα (λ = 0.71073) Mo Kα (λ = 0.71073) Mo Kα (λ = 0.71073 Mo Kα (λ = 0.71073) Cu Kα (λ = 1.54184) 6.38 to 59.06 8.46 to 147.42 2Θ range for data collection/° Index ranges 6.62 to 59.08 6.42 to 51.36 -14 ≤ h ≤ 10, -15 ≤ k ≤ -8 ≤ h ≤ 11, -25 ≤ k ≤ 15, -19 ≤ l ≤ 27 28, -14 ≤ l ≤ 14 4.74 to 60.72 -11 ≤ h ≤ 12, -23 ≤ k ≤ -24 ≤ h ≤ 20, -13 ≤ k ≤ -9 ≤ h ≤ 9, -38 ≤ k ≤ 40, 31, -14 ≤ l ≤ 12 14, -24 ≤ l ≤ 24 -13 ≤ l ≤ 13 Reflections collected 9366 6634 6999 19498 13120 Independent reflections 5515 [Rint = 0.0438, Rsigma = 0.0893] 3782 [Rint = 0.0263, Rsigma = 0.0397] 5022 [Rint = 0.0176, Rsigma = 0.0299] 7240 [Rint = 0.0499, Rsigma = 0.0645] 5776 [Rint = 0.0571, Rsigma = 0.0721] Data/restraints/para meters 5515/0/336 3782/2/316 7240/0/356 5776/0/363 5022/2/316 Goodness-of-fit on F2 1.040 1.047 1.022 1.051 1.064 Final R indexes [I>=2σ (I)] R1 = 0.0524, wR2 = 0.1092 R1 = 0.0228, wR2 = 0.0513 R1 = 0.0243, wR2 = 0.0614 R1 = 0.0499, wR2 = 0.0972 R1 = 0.0565, wR2 = 0.1339 Final R indexes [all data] R1 = 0.0636, wR2 = 0.1170 R1 = 0.0238, wR2 = 0.0519 R1 = 0.0258, wR2 = 0.0619 R1 = 0.0668, wR2 = 0.1063 R1 = 0.0707, wR2 = 0.1438 Largest diff. peak/hole / e Å-3 1.38/-0.90 0.47/-0.35 3.04/-2.54 2.14/-1.50 Flack parameter -0.07(5) 0.017(6) -- -- 0.78/-0.45 0.025(5) 2 Electronic Supplementary Information Table S 2 Selected bond angles (°) for complexes 1, 3 ( and ), 4a and 4b, s.u.s shown in parentheses Bonds 1 3 () 3 () 4a 4b X(1)-Rh(1)-X(2) X(1)-Rh(1)-O(2) X(2)-Rh(1)-O(2) X(1)-Rh(1)-N(1) X(2)-Rh(1)-N(1) X(1)-Rh(1)-N(2) X(2)-Rh(1)-N(2) X(1)-Rh(1)-N(3) X(2)-Rh(1)-N(3) N(1)-Rh(1)-O(2) N(2)-Rh(1)-O(2) N(3)-Rh(1)-O(2) N(1)-Rh(2)-N(2) N(1)-Rh(1)-N(3) N(2)-Rh(1)-N(3) 92.33(7) 177.09(15) 88.46(13) 89.41(17) 96.79(18) 90.46(18) 175.70(18) 96.85(19) 88.75(17) 93.3(2) 88.9(2) 80.4(2) 80.0(2) 171.5(2) 94.2(2) 93.00(4) 177.14(8) 86.00(8) 89.02(10) 96.06(10) 90.32(10) 175.11(9) 97.62(10) 86.14(10) 93.74(12) 90.85(13) 79.65(12) 80.40(14) 172.90(13) 96.98(13) 92.93(3) 177.19(7) 86.09(7) 89.25(9) 96.06(9) 90.50(9) 175.10(9) 97.66(9) 86.09(9) 93.47(11) 90.65(11) 79.66(11) 80.48(12) 172.67(12) 96.92(12) 90.86(5) 176.28(11) 91.14(11) 89.18(12) 95.31(13) 89.48(13) 175.45(12) 96.48(13) 88.26(13) 93.75(15) 88.78(16) 80.45(15) 80.16(17) 173.27(17) 96.21(17) 177.88(7) 86.76(16) 91.98(16) 175.3(2) 88.28(18) 90.58(19) 90.60(19) 92.26(18) 89.16(18) 97.7(2) 176.3(3) 78.4(2) 79.7(3) 175.3(2) 104.3(3) Table S 3 X-ray crystallographic data for complexes 6 and 7, s.u.s shown in parenthesis Complex 6 7 CCDC Number 1978079 1978083 Empirical formula C27H24Cl2I2N5O3Rh C27H24Br2I2N5O3Rh Formula weight 894.143 983.045 Temperature/K 100.0(3) 120.2(4) Crystal system monoclinic monoclinic Space group Cc Cc 15.5643(6) a/Å 15.4740(11) b/Å 9.1361(6) 9.1945(4) c/Å 21.4115(13) 21.4844(10) α/° 90 90 β/° 95.700(6) 95.650(4) γ/° 90 90 Volume/Å3 3012.0(4) 3059.6(2) Z 4 4 ρcalcg/cm3 1.972 2.134 μ/mm-1 2.832 5.225 F(000) 1714.2 1855.4 Crystal size/mm3 0.118 × 0.108 × 0.031 0.15 × 0.09 × 0.07 Radiation Mo Kα (λ = 0.71073) Mo Kα (λ = 0.71073) 2Θ range for data collection/° 6.22 to 52.74 6.18 to 62.3 Index ranges -16 ≤ h ≤ 19, -11 ≤ k ≤ 9, -26 ≤ l ≤ 21 -21 ≤ h ≤ 22, -11 ≤ k ≤ 12, -22 ≤ l ≤ 28 Reflections collected 9196 10526 Independent reflections 4805 [Rint = 0.0675, Rsigma = 0.1033] 6153 [Rint = 0.0368, Rsigma = 0.0602] Data/restraints/parameters 4805/98/309 6153/2/363 Goodness-of-fit on F2 1.052 1.018 Final R indexes [I>=2σ (I)] R1 = 0.0512, wR2 = 0.0906 R1 = 0.0383, wR2 = 0.0806 Final R indexes [all data] R1 = 0.0675, wR2 = 0.1016 R1 = 0.0432, wR2 = 0.0840 Largest diff. peak/hole / e Å-3 1.01/-0.91 1.00/-0.77 Flack parameter -0.06(4) 0.040(12) 3 Electronic Supplementary Information Table S 4 Selected bond angles (°) for complexes 6 and 7, s.u.s shown in parentheses Bonds 6 7 X(1)-Rh(1)-X(2) X(1)-Rh(1)-O(2) X(2)-Rh(1)-O(2) X(1)-Rh(1)-N(1) X(2)-Rh(1)-N(1) X(1)-Rh(1)-N(2) X(2)-Rh(1)-N(2) X(1)-Rh(1)-N(3) X(2)-Rh(1)-N(3) N(1)-Rh(1)-O(2) N(2)-Rh(1)-O(2) N(3)-Rh(1)-O(2) N(1)-Rh(2)-N(2) N(1)-Rh(1)-N(3) N(2)-Rh(1)-N(3) 177.34(5) 88.1(2) 89.4(2) 89.9(3) 89.5(3) 92.4(3) 90.0(3) 88.4(3) 91.9(3) 94.9(4) 174.9(4) 78.2(3) 80.0(4) 173.0(4) 106.7(5) 174.38(2) 88.21(14) 86.66(14) 89.13(16) 89.20(14) 91.31(16) 93.63(16) 92.10(15) 89.01(145 96.8(2) 175.7(2) 77.3(2) 79.0(2) 173.9(2) 107.0(2) 4 Electronic Supplementary Information Characterization by FTIR and PXRD FTIR Spectroscopy for Complex 3 Isomers Figure S 1 IR spectra of complexes 3a (red), 3b (orange) and 3c (yellow). PXRD of Complexes 3 and Complex 7 Figure S 2 Simulated and experimental PXRD diffractograms for a) complex 3 and b) complex 7. 5 Electronic Supplementary Information Characterization NMR Spectroscopy Complex 3 – 1st synthetic products Figure S 3 1H NMR spectra of complexes 3a (red) and 3c (yellow) (d6-acetone, 300 K, 300 MHz). Variable Temperature NMR Spectroscopy of Complex 3 – 1st synthetic product Figure S 4 Temperature-dependent 1H NMR spectra of complex 3c (d6-acetone, 300 K, 300 MHz). 6 Electronic Supplementary Information NMR for Complex 3 – 2nd synthetic product Figure S 5 Image of the combined crystals under a microscope Figure S 6 1H NMR spectra of complexes 3-combined (red) and 3-yellow microcrystals and 3-orange single crystals (d6-acetone, 300 K, 400 MHz). 7 Electronic Supplementary Information Figure S 7 1H NMR spectra of complexes 3-combined (red) and 3-yellow microcrystals and 3-orange single crystals (CDC3, 300 K, 400 MHz). 8 Electronic Supplementary Information NMR Characterisation and Exchange Studies of Complexes 1, 4, 5 and 8 Comparison NMR spectroscopy of complexes 1 and 5 1 H NMR spectra were recorded for [(3’-F)(3’-FH)RhCl2] (1, black) and [(3’-F)(3’-FH)RhI2] (5, blue) (Figure S4) at room temperature and show that complex 1 exhibits multiple isomers in solution, whereas complex 5 has only one isomer in solution. This can be clearly observed for the NH proton of the ligand, which appears at a chemical shift of ~12.5 ppm. Figure S 8 1H NMR spectrum of [(3’-F)(3’-FH)RhCl2] (1, black) and [(3’-F)(3’-FH)RhI2] (5, blue) confirming the different number of isomer present (CDCl3, 300 K, 400 MHz) 9 Electronic Supplementary Information NMR’s in Chloroform Complex 1 + N-(3-iodophenyl)picolinamide Scheme S1 Ligand exchange of [(3’-F)(3’-FH)RhCl2] (1) with ligand 3’-IH in CDCl3. Figure S 9 Overlay of the product (black) from the reaction of [(3’-F)(3’-FH)RhCl2] (1, dark green) + ligand 3’-IH (blue). The spectra for [(3’-I)(3’-IH)RhCl2] (4, dark blue) and ligand 3’-FH (dark red) are also included for comparison (CDCl3, 300 K, 400 MHz). The 1H NMR study of an equimolar mixture of [(3’-F)(3’-FH)RhCl2] (1) and N-(3-iodophenyl)picolinamide (3’-IH) in CDCl3 shows the formation of a mixed species which is possibly [(3’-F)(3’-IH)RhCl2] or [(3’-FH)(3’I)RhCl2] and the dissociation of N-(3-fluorophenyl)picolinamide (3’-FH). The resonances of [(3’-L)(3’LH)RhCl2] are “swamped” by the free ligand due to the complex's poor solubility in CDCl3. In addition, the 1H NMR spectra of [(3’-F)(3’-FH)RhCl2] (1) and [(3’-I)(3’-IH)RhCl2] (4) are essentially indistinguishable, thus inhibiting a direct in situ analysis of a mixed [(3’-F)(3’-IH)RhCl2] or [(3’-FH)(3’-I)RhCl2] complex. 10 Electronic Supplementary Information Complex 4 + N-(3-fluorophenyl)picolinamide Scheme S2 Attempted ligand exchange of [(3’-I)(3’-IH)RhCl2] (4) with 3’-FH in CDCl3. Figure S 10 Overlay of the product (black) from the reaction of [(3’-I)(3’-IH)RhCl2] (4, dark blue) + 3’-FH (dark red). The spectra for [(3’-F)(3’-FH)RhCl2] (1, dark green) and 3’-IH (blue) are also included for comparison (CDCl3, 300 K, 400 MHz). The 1H NMR study of an equimolar mixture of [(3’-I)(3’-IH)RhCl2] (4) and N-(3-fluorophenyl)picolinamide (3’-IH) in CDCl3 confirms that the N-(3-fluorophenyl)picolinamide is the ligand which is most easily substituted, and that the formation of the complex [(3’-I)(3’-IH)RhCl2] (4) is energetically favoured. The spectrum of the mixture shows essentially only the resonances of the added N-(3- fluorophenyl)picolinamide ligand, as the resonances of [(3’-X)(3’-XH)RhCl2] are again “swamped” by the free ligand due to the complex's poor solubility in CDCl3. No resonances of liberated N-(3fluorophenyl)picolinamide can be observed. 11 Electronic Supplementary Information Complex 5 + N-(3-iodophenyl)picolinamide Scheme S3 Test reaction for the ligand exchange between [(3’-F)(3’-FH)RhI2] (5) with 3’-IH in CDCl3. Figure S 11 Overlay of the product (black) from the reaction of [(3’-F)(3’-FH)RhI2] (5) + 3’-IH (blue) The spectra for ligand 3’-FH (dark red) is also included for comparison (CDCl3, 300 K, 400 MHz). The 1H-NMR study of an equimolar mixture of [(3’-F)(3’-FH)RhI2] (5) and N-(3-iodophenyl)picolinamide (3’-IH) in CDCl3 shows free 3’-IH exclusively. No ligand exchange can be observed as no resonances of liberated N-(3-fluorophenyl)picolinamide (3’-FH) are present. The extremely poor solubility of [(3’-F)(3’FH)RhI2] in CDCl3 does not allow for a direct comparison of [(3’-F)(3’-FH)RhI2] (5) and [(3’-I)(3’-IH)RhI2] (8) and a mixed complex [(3’-F)(3’-IH)RhI2] in this solvent. Therefore, the spectra for complex [(3’-I)(3’-IH)RhI2] (8) + 3’-FH was not recorded. 12 Electronic Supplementary Information NMR’s in Dimethyl sulfoxide Complex 1 + N-(3-iodophenyl)picolinamide Scheme S4 Test reaction for the ligand exchange between [(3’-F)(3’-FH)RhCl2] (1) with 3’-IH in d6-DMSO. Figure S 12 Overlay of the product (black) from the reaction of [(3’-F)(3’-FH)RhCl2] (1, dark green) + 3’-IH (blue). The spectra for ligand 3’-FH (dark red) is also included for comparison (d6-DMSO, 300 K, 400 MHz). The 1H-NMR study of an equimolar mixture of [(3’-F)(3’-FH)RhCl2] (1) and N-(3-iodophenyl)picolinamide (3’-IH) in d6-DMSO shows no changes in the resonances of the starting material and only an additional set of resonances for the free 3’-IH. No resonances of free 3’-FH can be observed, indicating no ligand exchange for [(3’-L)(3’-LH)RhCl2] complexes in DMSO. 13 Electronic Supplementary Information Complex 5 + N-(3-iodophenyl)picolinamide Scheme S5 Test reaction for the ligand exchange between [(3’-F)(3’-FH)RhI2] (5) with 3’-IH in d6-DMSO. Figure S 13 Overlay of the product (black) from the reaction of [(3’-F)(3’-FH)RhI2] (5) + 3’-IH (blue) The spectra for ligand 3’-FH (dark red) is also included for comparison (d6-DMSO, 300 K, 400 MHz). The 1H-NMR study of an equimolar mixture of [(3’-F)(3’-FH)RhI2] (5) and N-(3-iodophenyl)picolinamide (3’-IH) in d6-DMSO confirms that complexes of the type [(3’-L)(3’-LH)RhI2] do not undergo ligand exchange in DMSO. Along with the resonances of the starting material an additional set of resonances for the free 3’IH can be seen, with no resonances of free 3’-FH, further indicating a lack of ligand exchange in solution. 14 Electronic Supplementary Information Stability and Ligand Exchange Studies of Complex 3 and 7 over 96 h Figure S 14 Stability study of complex 3 in DMSO:PBS (80:20 v/v) over 96 h (d6-DMSO, 300 K, 400 MHz). 15 Electronic Supplementary Information Figure S 15 Stability study of complex 7 in MeCN:PBS (80:20 v/v) over 96 h (d3-MeCN, 300 K, 400 MHz). 16 Electronic Supplementary Information Figure S 16 Exchange studies of complex 3 with N-(3-chlorophenyl)picolinamide ligand (L’) in DMSO:PBS (80:20 v/v) over 96 h (d6-DMSO, 300 K, 400 MHz). 17 Electronic Supplementary Information Figure S 17 Overlay of complex 3 and 3’-BrH in MeCN a) without PBS, b) with PBS (80:20 v/v) after initial and c) with PBS (80:20 v/v) after 96 h (d3-MeCN, 300 K, 400 MHz). 18 Electronic Supplementary Information Figure S 18 Exchange studies of complex 7 with N-(3-chlorophenyl)picolinamide ligand (L’) in MeCN:PBS (80:20 v/v) over 96 h (d3-MeCN, 300 K, 400 MHz). 19 Electronic Supplementary Information Figure S 19 Overlay of complex 7 and 3’-BrH in MeCN a) without PBS, b) with PBS (80:20 v/v) after initial and c) with PBS (80:20 v/v) after 96 h (d3-MeCN, 300 K, 400 MHz). 20 Electronic Supplementary Information 96 h Chemosensitivity Studies Figure S 20 IC50 values (µM) ± SD of complexes 3, 7, 3-Ru and 7-Ru and CDDP against HCT116 p53+/+, HCT116 p53-/-, A549, FM55, MIA PaCa-2, A2780, A2780cisR, MCF-7 and ARPE-19. 24 and 72 h Chemosensitivity Studies Table S 5 IC50 values (µM) ± SD for complexes 3, 7, 3-Ru, 7-Ru and CDDP against HCT116 p53+/+, HCT116 p53-/-, and ARPE-19, after 24 h and 72 h incubation periods. Complexes HCT116 p53+/+ 3 24 h >100 72 h >100 7 1.7 ± 0.1 (21.8) 3-Ru 7-Ru CDDP 2.1 ± 0.2 (1.1) 1.7 ± 0.1 (1.6) 77 ± 2 (0.5) 1.33 ± 0.08 (27.8) 2.02 ± 0.08 (1.2) 1.29 ± 0.05 (2.1) 71.6 ± 0.4 (0.6) IC50 values (µM) ± SD HCT116 p53-/24 h 72 h >100 >100 ARPE-19 24 h >100 72 h >100 >100 (< 0.4) >100 (< 0.4) 37 ± 1 33 ± 1 2.3 ± 0.2 (1.0) 4.3 ± 0.1 (0.6) >100 (< 0.4) 2.10 ± 0.09 (1.1) 4.07 ± 0.07 (0.7) >100 (< 0.4) 2.41 ± 0.07 2.7 ± 0.2 41 ± 1 2.2 ± 0.3 2.5 ± 0.3 37 ± 1 21 Electronic Supplementary Information Selectivity Factors (SF) Table S 6 Selectivity Factors (SF) for complexes 7, 3-Ru, 7-Ru and CDDP when comparing the IC50 values of the isogenic colorectal cancer cell lines (HCT116 p53+/+ and HCT116 p53-/-) after 24, 72 and 96 h incubation periods. Complexes SF towards HCT116 p53-/- SF towards HCT116 p53+/+ 24 h 72 h 96 h 24 h 72 h 96 h 7 0.02 0.01 0.90 57.47 75.16 1.11 3-Ru 0.92 0.96 1.20 1.09 1.04 0.83 7-Ru 0.38 0.32 0.80 2.61 3.16 1.26 CDDP 0.77 0.72 0.42 1.30 1.40 2.36 LogP, Solubility and Pharmacokinetics Table S 7 Predictions of logP, solubility and pharmacokinetics for complexes 1-8, 3-Ru and 7-Ru, using Swiss ADME.1 Complex 1 Log P (MLOG) 3.54 LogS (ESOL) -8.33 Solubility mg/ mL 2.83x106 Class Poorly soluble GI absorption High 2 3.91 -8.75 1.16x106 Poorly soluble High 3 4 5 6 7 8 3-Ru 7-Ru 4.10 4.29 4.10 4.29 4.48 4.67 4.10 4.48 -9.38 -9.91 -9.09 -9.96 -10.59 -11.12 -9.37 -10.58 3.07x107 1.02x107 6.48x107 9.19x108 2.38x108 7.66x109 3.15x107 2.43x108 Poorly soluble Poorly soluble Poorly soluble Poorly soluble Insoluble Insoluble Poorly soluble Insoluble High High High High High High High Hugh Pharmacokinetics P-gp substrate, CYP1A2 inhibitor, CYP2C19 inhibitor BBB permeant, P-gp substrate, CYP3A4 inhibitor BBB permeant, P-gp substrate BBB permeant, P-gp substrate BBB permeant, P-gp substrate BBB permeant, P-gp substrate P-gp substrate BBB permeant, P-gp substrate BBB permeant, P-gp substrate P-gp substrate References (1) Daina, A.; Michielin, O.; Zoete, V. SwissADME: A Free Web Tool to Evaluate Pharmacokinetics, DrugLikeness and Medicinal Chemistry Friendliness of Small Molecules. Sci Rep 2017, 7 (Article number: 42717). https://doi.org/10.1038/srep42717. 22