The first structurally analysed nucleic acid building block containing the Reese protecting group: 2′-O-[1-(2-fluoro­phen­yl)-4-meth­oxy­piperidin-4-yl]-β-D-(1′R,2′R,3′R,4′R)-uridine

The title compound, C₂₁H₂₆FN₃O₇, is assembled by N—H...O and O—H...O hydrogen bonds into well-separated two-dimensional layers of about 15 Å thickness. The crescent conformation of the mol­ecules is stabilized by weak intra­molecular C—H...O and C—H...F hydrogen bonds. The uridine moiety adopts an anti conformation. The ribo­furan­ose ring exists in an envelope conformation. All the endocyclic uracil bonds are shorter than normal single C—N and C—C bonds, and five of them have comparable lengths, which implies a considerable degree of delocalization of the electron density within this ring.

S1. Introduction

Uridine, the one of four fundamental elements of RNA, is crucial in multiple biological processes, including coding, decoding, regulation and expression of genes. Structures of small uridine-containing species have been widely studied (mainly by spectroscopic techniques); however, only a very limited number of crystal structures studied by diffraction methods have been reported, probably as a result of problems with obtaining of such compounds in a form of single crystals. The Cambridge Structural Database (CSD, Version 5.35, with three updates; Groom & Allen, 2014) contains only 110 structures of compounds containing the uridine moiety with acyclic terminal O atoms of ribose, and 44 structures of compounds possessing at least one cyclic terminal O atom. The determination of structures of nucleotide building blocks (especially the unnatural ones) is of outstanding importance as it is crucial in the determination of the molecular mechanism of the action of biological molecules, in the design of new therapeutics [e.g. anti­biotics (Igarashi et al., 2005; Yamashita et al., 2003; Spork et al., 2010), anti­cancer agents (Hrdlicka et al., 2005), HIV inhibitors (Ivanova et al., 2007), regulators of gene expression (Murray et al., 2012; Obika et al., 2002)] and cellular transporting agents (Haziri & Leumann, 2012; Knoblauch et al., 1999). Up to today, only five such building blocks (with the 3'- and 5'-positions unsubstituted or substituted by phosphate moieties, and with an unsubstituted nucleobase) have been structurally studied by diffraction methods, namely: 2'-O-(4-meth­oxy­tetra­hydro­pyran-4-yl)uridine trihydrate (van Meervelt et al., 2004), 2'-O-(S)-tetra­hydro­pyran­yluridine (Lehmann et al., 1991), 2'-O-ribosyluridine (Markiewicz et al., 1998), (R_P)-2'-meth­oxy­uridin-3'-yl (N-isobutyryl 2'-meth­oxy­cytidin-5'-yl) methane­phosphono­thio­ate (Wozniak et al., 2004) and (+)-2'-O-(R)-tetra­hydro­pyran­yluridine (Stothart et al., 1973). The absence of substituents on the uracil ring of a building block is extremely important in the mild adjusting of RNA properties, because structural changes within the nucleobase often lead to distinctly different bonding (or even lack of bonding) of an RNA strand, and consequently a dysfunction of RNA (Spencer, 1972). On the other hand, the introduction of substituents at the 2'-position of uridine allows gentle tuning of RNA properties (Smith, 1998). The present report of 2'-O-[1-(2-Fluoro­phenyl)-4-meth­oxy­piperidin-4-yl]uridine, (I) (Fig. 1), is the first structurally analysed nucleic acid building block containing the Reese protecting group [1-(2-fluoro­phenyl)­piperidin-4-yl; Capaldi & Reese, 1994].

S2. Experimental

S2.1. Synthesis and crystallization

The general representation of synthesis procedure was depicted in Scheme 1. The preparation of the enanti­opure compound was based on methods reported in the literature (McGregor et al.,1996; Capaldi & Reese, 1994; Reese, 2005; Zhou et al.2007).

S2.1.1. Synthesis of 1-(2-fluoro­phenyl)-4-meth­oxy-1,2,5,6-tetra­hydro­pyridine

The acetal precursor was obtained with good yield by cyclization of 1,5-di­chloro­pentan-3-one and 2-fluoro­aniline via the Rees method. Elimination of methanol was achieved via a multi-step procedure. The acetal (24.15 g, 0.101 mol) was evaporated twice with dry toluene and dissolved in 151 ml of di­chloro­methane (DCM). The solution was placed in a three-necked round-bottomed septum-inlet flask equipped with a stirrer and a CaCl₂ tube. The reaction mixture was cooled to 273 K with a laminar flow of dry argon and then boron trifluoride di­ethyl etherate (24.7 ml) was added dropwise (continuously over a period of 3 m). The progress of the reaction (amount of acetal) was monitored via thin-layer chromatography (TLC; petroleum ether–ethyl acetate mixture, 7:1 v/v). After 2 h, all the acetal had reacted, the reaction mixture was warmed to ambient temperature and a saturated solution of aqueous NaHCO₃ (240 ml) was added. The layers were shaken vigorously for 20 min and separated using a separatory funnel. The aqueous layer was extracted twice with DCM and the combined organic layers were dried over anhydrous MgSO₄. The solvent was removed with a rotary evaporator and the solid residue was suspended in petroleum ether (150 ml). The suspension was filtered through a silica-gel path (Φ = 10 cm; h = 3 cm) on a sintered disc filter funnel. The silica gel was eluted with a petroleum ether–ethyl acetate mixture (98:2 v/v) until the product was absent in an eluate (detection on TLC under 254 nm UV lamp). The eluate was concentrated and subsequently evaporated twice with methanol. The resulting oil was cooled in a 2-propanol/CO₂(solid) bath to obtain a solid product. The solid product was evaporated twice with dry petroleum ether. The pale-yellow solid product was obtained in 95.4% yield (19.935 g). R_F = 0.70 (petroleum ether/AcOEt 7/1 v/v); ¹H NMR (250 MHz, CDCl₃, see Figs. S3–S4 in the Supporting information): δ 7.05–6.96 (m, 4H, aromatic), 4.71 (br t, 1H, J = 3 Hz), 3.70–3.66 (m, 2H), 3.57 (s, 3H, OCH₃), 3.31 (t, 2H, J = 6 Hz), 2.36–2.31 (m, 2H).

S2.1.2. \ Synthesis of 3',5'-O-(1,1,3,3-tetra­iso­propyl­disiloxan-1,3-diyl)-β-D-\ (1'R,2'R,3'R,4'R)-uridine

\ The β-D-uridine {1-[(3R,4S,5R)-3,4-di­hydroxy-5-(hy­droxy­methyl)­oxolan-2-\ yl]pyrimidine-2,4-dione; 6.0 g, 0.0246 mol) was evaporated twice with dry pyridine and dissolved in 130 ml of this solvent. 1,3-Di­chloro-1,1,3,3-tetra­iso­propyl­disiloxane (9.35 ml, 0.029 mol) was then added in one portion and the solution was stirred for 3 h. According to TLC analysis (chloro­form–methanol mixture, 95:5 v/v), after this time the whole substrate had reacted. Cold water (120 ml) was added to the reaction solution and the resulting mixture was extracted three times with DCM (150 ml). The combined organic extracts were washed with water (80 ml) and dried over anhydrous MgSO₄. The solvent was removed on a rotary evaporator and the crude residue was evaporated three times with toluene. The resulting stable foam was purified by column chromatography on silica gel (230–400 mesh) using isocratic elution (CHCl₃–MeOH 98:2 v/v). The final product was obtained in 89% yield (10.6 g of product). R_F = 0.43 (CHCl₃–MeOH 95/5 v/v); ¹H NMR (250 MHz, CDCl₃, see Figs. S5–S7 in the Supporting information): δ 8.49 (brs, 1H, NH), 7.67 (d, 1H, J = 8 Hz, H6), 5.73 (s, 1H, H1'), 5.69 (dd, 1H, J = 8 Hz, J = 2 Hz, H5), 4.38 (dd, 1H, J = 8.5 Hz, J = 4 Hz, H3'), 4.22–4.17 (m, 2H, H2', H5'), 4.10 (dt, 1H, J = 8.5 Hz, J = 3 Hz, H4'), 4.00 (dd, 1H, J = 13 Hz, J = 3 Hz, H5''), 3.00 (d, 1H, J = 1.2 Hz, 2'OH), 1.09–0.99 (m, 28H, i-Pr).

S2.1.3. \ Synthesis of 2'-O-[1-(2-fluoro­phenyl)-4-meth­oxy­piperidin-4-yl]-β-D-\ (1'R,2'R,3'R,4'R)-uridine, (I)

\ 3',5'-O-(1,1,3,3-Tetra­iso­propyl­disiloxan-1,3-diyl)-β-D-\ (1'R,2'R,3'R,4'R)-uridine (5 g, 0.0103 mol) was dissolved in freshly distilled DCM (41 ml) and the first portion of the enol ether (2.85 g, 0.0138 mol) and tri­fluoro­acetic acid (0.94 ml) was then added. After 24 h, the second portion of the enol ether (1.41 g, 0.0068 mol) was added. The solution was stirred for further 24 h, until TLC analysis (chloro­form–methanol 95:5 v/v) showed the absence of the substrate. The solution was alkalized with the tri­ethyl­amine and subsequently concentrated. The dark-brown–red oil was dissolved in a solution (1 mol dm^-3) of tetra­ethyl­ammonium fluoride in aceto­nitrile (30.6 ml, 0.0306 mol) and, after 1 h, total removal of the silyl protecting group was observed. The solution was concentrated, the residual oil was dissolved in chloro­form (75 ml) and a saturated solution (45 ml) of aqueous NaHCO₃ was added. The mixture was shaken vigorously for 35 min and during the shaking a pale-yellow solid precipitated. The mixture was placed in a refrigerator (at 277 K) for 40 min. The precipitate was filtered off and washed twice with water and di­ethyl ether. The white solid product was dried in a vacuum over CaCl₂. The product was obtained in 79.7% yield (3.7 g). R_F = 0.25 (CHCl₃/MeOH 95/5 v/v); ¹H NMR (250 MHz, DMSO-d₆): see Figs. S8–S9 in the Supporting information): δ 7.91 (d, 1H, J = 8 Hz, H6), 7.11–6.95 (m, 4H, aromatic fpmp), 6.01 (d, 1H, J = 7.75 Hz, H1'), 5.70 (d, 1H, J = 8 Hz, H5), 5.20 (brs, 2H, 3'OH, 5'OH), 4.35 (dd, 1H, J = 7.75, 4.75 Hz, H2'), 4.01–3.99 (m, 1H, H3''), 3.90 (brs, 1H, H4'), 3.56–3.55 (m, 2H, H5', H5''), 3.20–3.03 (m, 2H, piperidinyl), 2.98 (s, 3H, OCH₃), 2.88–2.69 (m, 2H, piperidinyl), 2.00–1.73 (m, 4H, piperidinyl); ¹⁹F NMR (235 MHz, DMSO-d₆, see Figs. S10—S11 in the Supporting information): δ -123.33 (s, 1F, product, 100%), -74.20 (s, residual CF₃COOH, 3%).

S2.1.4. Crystal preparation

Compound (I) (507 mg) was dissolved in CHCl₃ (14 ml). The crystallization vessel was transferred to a hermetic thermally isolated semiautomatic crystallizer and heated to the 334 K. The following parameters were applied to the crystallization process: temperature decreasing speed = 1 K per 24 h, vessel inter­nal volume enlargement = 0.5 ml per 24 h for the first 30 d of crystallization and 1 ml per 24 h for the next 10 d. After 40 d, the temperature was fixed at the 294 K and the vessel volume enlargement was set at 5 ml per 24 h. After the next 4 d, three small but relatively good quality crystals of (I) had grown. The crystals were removed from the solution using a Pasteur pipette, dried in a stream of dry helium and stored in sealed vessels.

S2.2. Refinement

Crystal data, data collection and structure refinement details are summarized in Table 1. H atoms were placed in calculated positions (C—H = 0.93, 0.96, 0.97 or 0.98 Å, respectively, for aromatic, methyl, methane­diyl and methane­triyl C atoms, O—H = 0.82 Å and N—H = 0.86 Å) and were refined as riding on their adjacent atoms, with fixed U_iso values [U_iso(H) = 1.2U_eq(C) for non-methyl C atoms and 1.5U_eq(C,O,N) otherwise] in all (including the final) refinement cycles. The methyl and hyd­oxy groups were allowed to rotate along preceding C—C and C—O bonds. The isotropic displacement parameters of O- and N-bonded H atoms were then refined to check the correctness of their positions in the post-final calculation. After eight cycles, the refinement reached stable convergence with isotropic displacement parameters in the range 0.03–0.09 Ų. The values of the isotropic displacement parameters of the H atoms have reasonable values (in comparison to the parent non-H atoms), which proves correctness of the H-atom positions.

S3. Results and discussion

\ The conformation of ribonucleosides is characterized by three parameters: the O_endocyclic—C—N_nucleobase—C_carbonyl torsion angle (determining the relative conformation of a nitro­genous base and a sugar in syn–anti terms), the sugar ring puckering (described in terms of pseudorotation cycle) and the C_endocyclic—C_exocyclic bond orientation towards the sugar ring (Blackburn et al., 2006). In (I), the O1—C1—N1—C6 torsion angle is -127.43 (18)°, what means that in (I) the uridine moiety adopts a more common anti conformation (Blackburn et al., 2006). There are known two compounds with the O_endocyclic—C—N_nucleobase—C_carbonyl torsion angle close to the value seen in (I), i.e. 128.6 and 126.7° for tri­ethyl­ammonium uridine-3'-O-thio­phosphate methyl ester (Saenger et al., 1974) and 2'-C-eth­oxy-3',5'-O-(tetra­iso­propyl­disiloxane-1,3-diyl)uridine (Chi et al., 2001), respectively. In (I), atoms C2 and C3 atoms deviate from the weighted least-squares plane calculated through C1/O1/C4 by -0.637 (5) and 0.010 (5) Å, respectively; thus, the conformation of the five-membered ring can be described as C^2'-endo, with a pseudorotation phase angle P of 162.1 (2)° (Rao et al., 1981). This is an envelope conformation with the C2 atom at the flap [asymmetry parameters (Duax & Norton, 1975): ΔC_s(C2,C4–O4) = 0.80 (18)°, ΔC₂(O4,C2—C3) = 20.79 (19)°]. The deviations of atoms C2 and C3 from the above-mentioned plane is correlated in known compounds (included in the CSD; Groom & Allen, 2014) and it can be expressed by the equation d_{C1/O1/C4–C3} = d_{C1/O1/C4—C2} ± 0.6 (see Fig. S1 in the Supporting information). The values of (I) fit well to the presented equation. The C_endocyclic—C_exocyclic bond direct environment adopts the sc⁺ conformation which frequently exists in uridine compounds. There are five compounds with the O1—C4—C5—O4 torsion angle close to the value in compound (I) [-72.4 (2)°], namely: 6-tert-butyl-1-[3,4-di­hydroxy-5-(hy­droxy­methyl)­tetra­hydro­furan-2-yl]\ pyrimidine-2,4(1H,3H)-dione (Shih et al., 2013), pure uridine (Green et al., 1975), 5-acetyl-1-(3,5-O-iso­propyl­idene-b-D-xylo­furan­osyl)uracil (Jones & Sowden, 1982), 2,2'-di­pyridyl­amine­(uridine-5'-phosphato)copper(II) dimer penta­hydrate (Fischer & Bau, 1978) and 2'-O-(S)-tetra­hydro­pyran­yluridine (Lehmann et al., 1991) with angles of 72.18, 72.97, 72.93, 72.76 and and 72.20°, respectively. Atoms N1 and C2 occupy equatorial positions and atoms O3 and C5 occupy axial positions of the ribo­furan­ose ring, and all the chiral centres have the same R configuration. The absolute configuration of (I) cannot be determined on the basis of the anomalous scattering because, in the studied case, the anomalous-dispersion effects are insignificant (Flack & Bernardinelli, 1999, 2000); however, the chiral centres of the substrate (1-[(3R,4S,5R)-3,4-di­hydroxy-5-(hy­droxy­methyl)­oxolan-2-yl]\ pyrimidine-2,4-dione) were unchanged in the synthesis and (I) is enanti­omerically pure and contains only a β-D-ribo­furan­ose ring. The enanti­opurity of (I) was confirmed by analysis of the ¹H NMR spectra (see Figs. S3–S9 in the Supporting information), i.e. none of the spectra show splitting or doubling of the signals (doublets) of ribo­furan­ose ring H atoms, as well as by liquid chromatography, i.e. transferring of the solution of (I) through a chiral column [(S,S)-Whelk-O 1, particle size 100 Å, column length × inter­nal diameter: 25 × 0.46 cm] does not lead to separation of (I) into the two or more substances (if the chiral centres configuration is not retained, at least two different species must be present in the eluate; Magora et al., 2000). The piperidine moiety adopts almost ideal chair conformation with the atom placed ΔC_s asymmetry parameters (Duax & Norton, 1975) in range 0.6–3.9° and, bond mid-points crossing ΔC₂ asymmetry parameters in range 2.8–5.4°. All the endocyclic uracil bonds are shorter than the single C—N and C—C bonds and five of them (except for the C8—C7 bond) have comparable lengths, which implies a considerable degree of delocalization of the electron density within this ring. The uracil and the 2-fluoro­phenyl rings are planar in the range of experimental error and the 2-fluoro­phenyl moiety is almost coplanar with the N3—C13—C14 part of the piperidine ring.

The molecules of (I) are held together by N—H···O and O—H···O inter­molecular hydrogen bonds (Table 2). These hydrogen bonds together create C(6)C(10)C(7) unitary graph set (Etter, 1990, 1991; Etter et al., 1990; Bernstein, 1991; Bernstein et al., 1990, 1995; see Fig. S2 in the Supporting information). At the second level graph, N₂, the hydrogen bonds form complex motifs including the C₂²(9)C₂²(9)C₂²(12)C₂²(14)\ C₂²(15)C₂²(16)C₂²(16) ones of the basic graph sets. In this way, a layer (of about 15 Å thickness) parallel to the crystallographic (001) plane is created (Fig. 2). Neighbouring layers are well separated and are not connected by classical or weak hydrogen bonds. Each molecule of (I) contains C—H···O and C—H···F short contacts (Table 2) which can be classified as weak intra­molecular hydrogen bonds (Desiraju & Steiner, 1999). The short C13—H13B···Cg contact [Cg is the centroid of the ring containing atoms N1/N2/C6/C7/C8/C9; H···Cg = 2.73 Å, C···Cg = 3.594 (3) Å and C—H···Cg = 149.0 °] can also be considered as an intra­molecular C—H···π inter­action. The possibility of the formation of π–π stacking inter­actions was rejected on the basis of the large delocalized ring-centroid distances [the shortest is 4.8224 (13) Å] accompanied by the large inclination of these rings (range 34–68 °).

Computing details

Data collection: CrysAlis CCD (UNIL IC & Kuma, 2000); cell refinement: CrysAlis RED (UNIL IC & Kuma, 2000); data reduction: CrysAlis RED (UNIL IC & Kuma, 2000); program(s) used to solve structure: SHELXS97 (Sheldrick, 2008); program(s) used to refine structure: SHELXL2014 (Sheldrick, 2015); molecular graphics: XP in SHELXTL/PC (Bruker, 2003) ORTEP-3 for Windows (Farrugia, 2012); software used to prepare material for publication: SHELXL2014 (Sheldrick, 2015) and PLATON (Spek, 2009).

Figures

	Fig. 1. A view of the asymmetric unit of (I), showing the atom-labelling scheme. Displacement ellipsoids are drawn at the 50% probability level and H atoms are shown as spheres of arbitrary radii.
	Fig. 2. A part of the molecular packing of (I), showing the hydrogen-bonded layers parallel to the crystallographic (001) plane. Intermolecular hydrogen bonds are indicated by dashed lines.

2'-O-[1-(2-Fluorophenyl)-4-methoxypiperidin-4-yl]–β-D-(1'R,2'R,3'R,4'R)-uridine

Crystal data

C21H26FN3O7	F(000) = 476
Mr = 451.45	Dx = 1.440 Mg m^−3 Dm = 1.44 Mg m^−3 Dm measured by Berman density torsion balance
Monoclinic, P21	Mo Kα radiation, λ = 0.71073 Å
a = 10.5197 (5) Å	Cell parameters from 2730 reflections
b = 6.7783 (4) Å	θ = 4–20°
c = 15.0390 (7) Å	µ = 0.11 mm^−1
β = 103.807 (4)°	T = 290 K
V = 1041.38 (9) Å^3	Prism, colourless
Z = 2	0.45 × 0.36 × 0.35 mm

Data collection

Kuma KM-4-CCD diffractometer	3334 independent reflections
Radiation source: fine-focus sealed tube	3300 reflections with I > 2σ(I)
Graphite monochromator	Rint = 0.034
Detector resolution: 12.8 pixels mm^-1	θmax = 25.1°, θmin = 3.6°
ω scans	h = −12→12
Absorption correction: numerical (X-RED; Stoe & Cie, 1999)	k = −8→6
Tmin = 0.948, Tmax = 0.965	l = −17→17
11720 measured reflections

Refinement

Refinement on F^2	Hydrogen site location: inferred from neighbouring sites
Least-squares matrix: full	H-atom parameters constrained
R[F^2 > 2σ(F^2)] = 0.028	w = 1/[σ^2(Fo^2) + (0.0332P)^2 + 0.1961P] where P = (Fo^2 + 2Fc^2)/3
wR(F^2) = 0.071	(Δ/σ)max = 0.001
S = 1.08	Δρmax = 0.14 e Å^−3
3334 reflections	Δρmin = −0.14 e Å^−3
293 parameters	Extinction correction: SHELXL2014 (Sheldrick, 2015), Fc^*=kFc[1+0.001xFc^2λ^3/sin(2θ)]^-1/4
1 restraint	Extinction coefficient: 0.015 (2)
Primary atom site location: structure-invariant direct methods	Absolute structure: Flack x determined using 1279 quotients [(I+)-(I-)]/[(I+)+(I-)] (Parsons et al., 2013).
Secondary atom site location: difference Fourier map	Absolute structure parameter: −0.3 (3)

Special details

Geometry. All e.s.d.'s (except the e.s.d. in the dihedral angle between two l.s. planes) are estimated using the full covariance matrix. The cell e.s.d.'s are taken into account individually in the estimation of e.s.d.'s in distances, angles and torsion angles; correlations between e.s.d.'s in cell parameters are only used when they are defined by crystal symmetry. An approximate (isotropic) treatment of cell e.s.d.'s is used for estimating e.s.d.'s involving l.s. planes.

Fractional atomic coordinates and isotropic or equivalent isotropic displacement parameters (Ų)

	x	y	z	Uiso*/Ueq
O1	0.71232 (13)	0.0943 (2)	0.94784 (10)	0.0315 (3)
C1	0.74581 (18)	0.2458 (3)	0.89190 (13)	0.0254 (4)
H1A	0.7340	0.3752	0.9179	0.031*
C2	0.64835 (18)	0.2215 (3)	0.79982 (13)	0.0242 (4)
H2A	0.6721	0.1074	0.7671	0.029*
C3	0.52308 (19)	0.1784 (3)	0.83084 (14)	0.0280 (4)
H3A	0.4578	0.1115	0.7829	0.034*
C4	0.57391 (19)	0.0432 (3)	0.91355 (14)	0.0276 (4)
H4A	0.5273	0.0749	0.9609	0.033*
C5	0.5601 (2)	−0.1740 (4)	0.89337 (15)	0.0371 (5)
H5A	0.6112	−0.2481	0.9448	0.044*
H5B	0.4691	−0.2127	0.8843	0.044*
O2	0.64756 (13)	0.3954 (2)	0.74734 (9)	0.0269 (3)
O3	0.47449 (15)	0.3593 (2)	0.85679 (12)	0.0393 (4)
H3O	0.4069	0.3388	0.8735	0.059*
O4	0.6043 (2)	−0.2166 (3)	0.81369 (13)	0.0564 (5)
H4O	0.5990	−0.3357	0.8038	0.085*
N1	0.88232 (15)	0.2199 (3)	0.88946 (11)	0.0267 (4)
C6	0.96560 (19)	0.3804 (3)	0.91144 (13)	0.0281 (4)
N2	1.09408 (17)	0.3399 (3)	0.91195 (12)	0.0334 (4)
H2N	1.1492	0.4341	0.9293	0.050*
C7	1.1442 (2)	0.1657 (4)	0.88777 (16)	0.0385 (6)
C8	1.0500 (2)	0.0103 (4)	0.86153 (17)	0.0388 (6)
H8A	1.0752	−0.1108	0.8422	0.047*
C9	0.9254 (2)	0.0414 (3)	0.86509 (15)	0.0314 (5)
H9A	0.8659	−0.0620	0.8505	0.038*
O5	0.93010 (16)	0.5440 (3)	0.92739 (13)	0.0421 (4)
O6	1.26199 (16)	0.1530 (4)	0.89020 (14)	0.0577 (6)
C10	0.6329 (2)	0.3732 (4)	0.64967 (14)	0.0323 (5)
C11	0.6332 (2)	0.5837 (4)	0.61425 (15)	0.0358 (5)
H11A	0.6093	0.5827	0.5478	0.043*
H11B	0.5684	0.6610	0.6353	0.043*
C12	0.7669 (2)	0.6785 (4)	0.64731 (16)	0.0360 (5)
H12A	0.7651	0.8100	0.6215	0.043*
H12B	0.7869	0.6911	0.7134	0.043*
N3	0.87058 (19)	0.5620 (3)	0.62092 (12)	0.0344 (4)
C13	0.8754 (2)	0.3623 (4)	0.65829 (17)	0.0385 (5)
H13A	0.8983	0.3684	0.7246	0.046*
H13B	0.9426	0.2871	0.6391	0.046*
C14	0.7447 (2)	0.2595 (4)	0.62597 (16)	0.0399 (6)
H14A	0.7499	0.1297	0.6537	0.048*
H14B	0.7265	0.2421	0.5601	0.048*
C15	0.9934 (2)	0.6599 (4)	0.63805 (15)	0.0356 (5)
C16	1.0078 (2)	0.8297 (4)	0.58904 (15)	0.0410 (6)
C17	1.1237 (3)	0.9295 (5)	0.59936 (19)	0.0516 (7)
H17A	1.1284	1.0428	0.5654	0.062*
C18	1.2332 (3)	0.8592 (5)	0.6610 (2)	0.0585 (8)
H18A	1.3128	0.9248	0.6689	0.070*
C19	1.2242 (3)	0.6923 (5)	0.71063 (18)	0.0542 (7)
H19A	1.2978	0.6453	0.7525	0.065*
C20	1.1063 (2)	0.5931 (5)	0.69892 (17)	0.0452 (6)
H20A	1.1025	0.4791	0.7326	0.054*
F1	0.90177 (15)	0.8989 (3)	0.52599 (11)	0.0564 (4)
O7	0.51962 (16)	0.2638 (3)	0.61102 (11)	0.0426 (4)
C21	0.3978 (3)	0.3601 (5)	0.6091 (2)	0.0562 (7)
H21A	0.3868	0.4696	0.5674	0.084*
H21B	0.3977	0.4068	0.6693	0.084*
H21C	0.3272	0.2684	0.5891	0.084*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
O1	0.0252 (7)	0.0359 (9)	0.0324 (7)	−0.0034 (6)	0.0049 (6)	0.0086 (7)
C1	0.0230 (9)	0.0249 (11)	0.0288 (9)	−0.0003 (8)	0.0069 (7)	−0.0006 (8)
C2	0.0249 (9)	0.0196 (11)	0.0277 (9)	−0.0015 (8)	0.0053 (7)	0.0001 (8)
C3	0.0223 (9)	0.0252 (11)	0.0360 (10)	0.0001 (8)	0.0060 (8)	0.0001 (9)
C4	0.0225 (9)	0.0295 (12)	0.0314 (10)	−0.0021 (8)	0.0080 (8)	0.0002 (9)
C5	0.0440 (13)	0.0283 (13)	0.0390 (11)	−0.0059 (10)	0.0102 (9)	0.0045 (10)
O2	0.0337 (7)	0.0202 (8)	0.0273 (7)	−0.0011 (6)	0.0082 (5)	0.0007 (6)
O3	0.0337 (8)	0.0283 (10)	0.0603 (10)	0.0045 (7)	0.0199 (7)	0.0002 (8)
O4	0.1009 (16)	0.0241 (10)	0.0526 (10)	0.0033 (10)	0.0349 (10)	−0.0026 (8)
N1	0.0206 (8)	0.0264 (11)	0.0323 (9)	−0.0023 (7)	0.0047 (6)	−0.0018 (7)
C6	0.0261 (9)	0.0286 (13)	0.0285 (10)	−0.0037 (9)	0.0044 (7)	−0.0011 (9)
N2	0.0237 (8)	0.0351 (11)	0.0406 (9)	−0.0080 (8)	0.0059 (7)	−0.0028 (8)
C7	0.0279 (11)	0.0478 (16)	0.0411 (12)	−0.0017 (10)	0.0110 (9)	−0.0038 (11)
C8	0.0291 (11)	0.0386 (15)	0.0492 (13)	0.0024 (10)	0.0100 (9)	−0.0107 (11)
C9	0.0279 (10)	0.0264 (13)	0.0395 (11)	−0.0035 (9)	0.0073 (8)	−0.0056 (9)
O5	0.0378 (9)	0.0281 (10)	0.0585 (10)	−0.0049 (7)	0.0078 (7)	−0.0095 (8)
O6	0.0264 (8)	0.0706 (15)	0.0802 (13)	−0.0037 (9)	0.0206 (8)	−0.0168 (11)
C10	0.0405 (11)	0.0297 (13)	0.0262 (10)	−0.0058 (10)	0.0070 (8)	−0.0017 (9)
C11	0.0440 (13)	0.0311 (13)	0.0326 (10)	0.0012 (10)	0.0096 (9)	0.0060 (10)
C12	0.0477 (13)	0.0221 (12)	0.0422 (12)	0.0014 (10)	0.0186 (10)	0.0000 (10)
N3	0.0452 (11)	0.0244 (11)	0.0376 (9)	−0.0016 (8)	0.0180 (8)	0.0008 (8)
C13	0.0507 (13)	0.0241 (13)	0.0483 (13)	0.0040 (11)	0.0266 (10)	0.0036 (10)
C14	0.0590 (15)	0.0238 (12)	0.0436 (12)	−0.0034 (11)	0.0258 (11)	−0.0038 (10)
C15	0.0447 (12)	0.0332 (14)	0.0322 (11)	−0.0020 (10)	0.0157 (9)	−0.0024 (10)
C16	0.0463 (13)	0.0411 (15)	0.0352 (11)	−0.0062 (11)	0.0087 (10)	0.0023 (10)
C17	0.0582 (16)	0.0457 (18)	0.0526 (15)	−0.0174 (13)	0.0164 (12)	0.0036 (13)
C18	0.0488 (15)	0.064 (2)	0.0599 (17)	−0.0194 (15)	0.0082 (12)	−0.0088 (16)
C19	0.0450 (14)	0.067 (2)	0.0473 (14)	0.0016 (14)	0.0037 (11)	−0.0030 (14)
C20	0.0505 (14)	0.0440 (16)	0.0421 (13)	0.0041 (12)	0.0129 (10)	0.0047 (12)
F1	0.0526 (9)	0.0545 (11)	0.0576 (9)	−0.0101 (7)	0.0039 (7)	0.0245 (8)
O7	0.0451 (9)	0.0391 (10)	0.0390 (8)	−0.0114 (8)	0.0010 (7)	−0.0052 (7)
C21	0.0415 (13)	0.059 (2)	0.0600 (16)	−0.0081 (14)	−0.0042 (11)	0.0051 (15)

Geometric parameters (Å, º)

O1—C1	1.424 (3)	C10—C14	1.519 (3)
O1—C4	1.465 (2)	C10—C11	1.523 (3)
C1—N1	1.456 (2)	C11—C12	1.519 (3)
C1—C2	1.523 (3)	C11—H11A	0.9700
C1—H1A	0.9800	C11—H11B	0.9700
C2—O2	1.417 (2)	C12—N3	1.476 (3)
C2—C3	1.527 (3)	C12—H12A	0.9700
C2—H2A	0.9800	C12—H12B	0.9700
C3—O3	1.419 (3)	N3—C15	1.420 (3)
C3—C4	1.535 (3)	N3—C13	1.462 (3)
C3—H3A	0.9800	C13—C14	1.514 (3)
C4—C5	1.503 (3)	C13—H13A	0.9700
C4—H4A	0.9800	C13—H13B	0.9700
C5—O4	1.415 (3)	C14—H14A	0.9700
C5—H5A	0.9700	C14—H14B	0.9700
C5—H5B	0.9700	C15—C20	1.391 (3)
O2—C10	1.448 (2)	C15—C16	1.394 (4)
O3—H3O	0.8200	C16—F1	1.363 (3)
O4—H4O	0.8200	C16—C17	1.371 (4)
N1—C9	1.372 (3)	C17—C18	1.380 (4)
N1—C6	1.387 (3)	C17—H17A	0.9300
C6—O5	1.212 (3)	C18—C19	1.371 (5)
C6—N2	1.377 (3)	C18—H18A	0.9300
N2—C7	1.377 (3)	C19—C20	1.385 (4)
N2—H2N	0.8600	C19—H19A	0.9300
C7—O6	1.234 (3)	C20—H20A	0.9300
C7—C8	1.435 (3)	O7—C21	1.432 (3)
C8—C9	1.342 (3)	C21—H21A	0.9600
C8—H8A	0.9300	C21—H21B	0.9600
C9—H9A	0.9300	C21—H21C	0.9600
C10—O7	1.406 (3)
C1—O1—C4	109.12 (14)	O7—C10—C11	114.94 (18)
O1—C1—N1	108.21 (16)	O2—C10—C11	104.43 (17)
O1—C1—C2	104.55 (16)	C14—C10—C11	109.09 (19)
N1—C1—C2	114.71 (16)	C12—C11—C10	111.16 (19)
O1—C1—H1A	109.7	C12—C11—H11A	109.4
N1—C1—H1A	109.7	C10—C11—H11A	109.4
C2—C1—H1A	109.7	C12—C11—H11B	109.4
O2—C2—C1	109.23 (16)	C10—C11—H11B	109.4
O2—C2—C3	115.98 (16)	H11A—C11—H11B	108.0
C1—C2—C3	100.70 (15)	N3—C12—C11	111.8 (2)
O2—C2—H2A	110.2	N3—C12—H12A	109.3
C1—C2—H2A	110.2	C11—C12—H12A	109.3
C3—C2—H2A	110.2	N3—C12—H12B	109.3
O3—C3—C2	108.22 (17)	C11—C12—H12B	109.3
O3—C3—C4	111.84 (17)	H12A—C12—H12B	107.9
C2—C3—C4	101.68 (15)	C15—N3—C13	114.6 (2)
O3—C3—H3A	111.6	C15—N3—C12	113.55 (19)
C2—C3—H3A	111.6	C13—N3—C12	110.65 (17)
C4—C3—H3A	111.6	N3—C13—C14	110.9 (2)
O1—C4—C5	110.03 (18)	N3—C13—H13A	109.5
O1—C4—C3	105.60 (16)	C14—C13—H13A	109.5
C5—C4—C3	115.02 (18)	N3—C13—H13B	109.5
O1—C4—H4A	108.7	C14—C13—H13B	109.5
C5—C4—H4A	108.7	H13A—C13—H13B	108.0
C3—C4—H4A	108.7	C13—C14—C10	112.7 (2)
O4—C5—C4	109.62 (19)	C13—C14—H14A	109.0
O4—C5—H5A	109.7	C10—C14—H14A	109.0
C4—C5—H5A	109.7	C13—C14—H14B	109.0
O4—C5—H5B	109.7	C10—C14—H14B	109.0
C4—C5—H5B	109.7	H14A—C14—H14B	107.8
H5A—C5—H5B	108.2	C20—C15—C16	115.6 (2)
C2—O2—C10	117.63 (15)	C20—C15—N3	124.4 (2)
C3—O3—H3O	109.5	C16—C15—N3	119.9 (2)
C5—O4—H4O	109.5	F1—C16—C17	117.8 (2)
C9—N1—C6	121.70 (17)	F1—C16—C15	118.5 (2)
C9—N1—C1	120.24 (17)	C17—C16—C15	123.6 (2)
C6—N1—C1	118.05 (18)	C16—C17—C18	118.9 (3)
O5—C6—N2	122.0 (2)	C16—C17—H17A	120.6
O5—C6—N1	123.92 (18)	C18—C17—H17A	120.6
N2—C6—N1	114.10 (19)	C19—C18—C17	119.7 (3)
C6—N2—C7	127.33 (19)	C19—C18—H18A	120.2
C6—N2—H2N	116.3	C17—C18—H18A	120.2
C7—N2—H2N	116.3	C18—C19—C20	120.5 (3)
O6—C7—N2	119.8 (2)	C18—C19—H19A	119.7
O6—C7—C8	125.4 (3)	C20—C19—H19A	119.7
N2—C7—C8	114.78 (19)	C19—C20—C15	121.6 (3)
C9—C8—C7	119.4 (2)	C19—C20—H20A	119.2
C9—C8—H8A	120.3	C15—C20—H20A	119.2
C7—C8—H8A	120.3	C10—O7—C21	115.7 (2)
C8—C9—N1	122.5 (2)	O7—C21—H21A	109.5
C8—C9—H9A	118.8	O7—C21—H21B	109.5
N1—C9—H9A	118.8	H21A—C21—H21B	109.5
O7—C10—O2	110.41 (16)	O7—C21—H21C	109.5
O7—C10—C14	105.2 (2)	H21A—C21—H21C	109.5
O2—C10—C14	112.94 (17)	H21B—C21—H21C	109.5
O1—C1—N1—C6	−127.43 (18)	O1—C4—C5—O4	−72.4 (2)
C4—O1—C1—N1	−148.28 (16)

Hydrogen-bond geometry (Å, º)

D—H···A	D—H	H···A	D···A	D—H···A
N2—H2N···O1^i	0.86	2.33	3.085 (2)	147
O3—H3O···O6^ii	0.82	2.04	2.782 (2)	151
O4—H4O···O2^iii	0.82	2.12	2.888 (2)	155
C1—H1A···O5	0.98	2.33	2.763 (3)	106
C2—H2A···O4	0.98	2.46	3.020 (3)	116
C12—H12A···F1	0.97	2.34	2.968 (3)	122

Symmetry codes: (i) −x+2, y+1/2, −z+2; (ii) x−1, y, z; (iii) x, y−1, z.