N-[tert-Butoxy­carbonyl­glycyl-(Z)-α,β-dehydro­phenyl­alanylglycyl-(E)-α,β-dehydro­phenyl­alanylphenyl­alanyl]-4-nitro­aniline ethanol solvate

The α,β-dehydro­phenyl­alanine residues influence the conformation of the title penta­peptide Boc⁰–Gly¹–Δ^ZPhe²–Gly³–Δ^EPhe⁴–L-Phe⁵–p-NA ethanol solvate, C₄₂H₄₃N₇O₉·C₂H₅OH. The first unsaturated phenyl­alanyl (Δ^ZPhe²) and the third glycyl (Gly³) residues form a type I β turn, while the second unsaturated phenyl­alanyl (Δ^EPhe⁴) and the last phenyl­alanyl (L-Phe⁵) residues are part of a type II β turn. All the amino acids in the peptide are linked trans to one another. The crystal structure is stabilized by intra- and inter­molecular hydrogen bonds.

S1. Comment

α,β-Dehydroamino acid residues (amino acids with a double bond between the C_α and C_β atoms) have been found in many biologically active peptides having antibiotic properties (Noda et al., 1983). Incorporation of a dehydroamino acid into a peptide decreases conformational flexibility (Aubry et al., 1984). The molecular structures of α,β-dehydrophenylalanine-containing (ΔPhe) peptides have shown that α,β-dehydrophenylalanine induces β turns (Venkatachalam, 1968) in short sequences with one ΔPhe residue (Główka et al., 1987; Główka, 1988) and a 3₁₀ helical conformation in longer sequences (Rajashankar et al., 1992; Rajashankar, Ramakumar, Jain et al., 1995; Rajashankar, Ramakumar, Mal et al., 1995; Padmanabhan & Singh, 1993; Jain et al., 1997). The number and position of ΔPhe residues and the type of neighbouring amino acids also play an important role in peptide chain conformation (Rajashankar et al., 1996).

The present paper reports the crystal structure of the title pentapeptide Boc⁰—Gly¹–Δ^ZPhe²—Gly³–Δ^EPhe⁴—L-Phe⁵–p-NA ethanol solvate, (I) (p-NA is para-nitroaniline), containing one Δ^ZPhe (isomer Z of an α,β-dehydrophenylalanine residue, i.e. with the aromatic ring cis to the N atom) between two flexible glycine residues and one Δ^EPhe (isomer E of an α,β-dehydrophenylalanine residue, i.e. with the aromatic ring trans to the N atom) between glycine and phenylalanine residues. There is one molecule in the asymmetric part of the unit cell. The atom-numbering scheme and a general view of the molecule are shown in Fig. 1, while selected bond lengths and angles are given in Table 1.

The C_α═C_β (C8═C9 and C19═C20) distances for the ΔPhe residues of (I) are in agreement with those found in other structures containing ΔPhe (Główka, 1988). A shortening of about 0.045 (7) Å for the N2—C_α8 bond in Δ^ZPhe² and 0.041 (7) Å for the N4—C_α19 bond in Δ^EPhe⁴ is observed with respect to the corresponding bonds in the saturated Phe⁵ residue (N5—C_α28). The torsion angles χ² [6.3 (11)°], χ^2,1 [22.3 (11)°] and χ^2,2 [−160.4 (7)°] of the Δ^ZPhe² residue suggest that its side chain is almost planar. The torsion angles χ⁴ [−172.3 (6)°], χ^4,1 [38.3 (11)°] and χ^4,2 [−144.2 (7)°] of the Δ^EPhe⁴ residue suggest that, in this case, the side chain is antiperiplanar (Table 1). Are definitions needed for these χ angles? The steric contacts between the side-chain and main-chain atoms of Δ^ZPhe² and Δ^EPhe⁴ are partially relaxed by rearrangement of the bond angles at the C_α and C_β atoms of these residues. As in other cases (Pieroni et al., 1975, 1976/77; Aubry et al., 1985), the cinnamic moieties of the ΔPhe residues in the title peptide are not planar. The torsion angles between the C═C and C═O bonds of Δ^ZPhe² and Δ^EPhe⁴ are 29.1 (9) and −113.2 (8)°, respectively.

All the amino acids in the title pentapeptide are linked trans to each other. The deviations of all ω angles are not larger than 9°. The values of torsion angles ϕ and ψ of the Δ^ZPhe² and Gly³ residues suggest a type I β turn conformation, while the torsion angles of Δ^EPhe⁴ and Phe⁵ indicate that these residues form a type II β turn. The torsion angles of the tert-butoxycarbonyl group (ϕ⁰ and ω⁰) correspond to a trans–trans conformation.

The conformation of (I) is stabilized by nine intramolecular and four intermolecular hydrogen bonds of different types, namely N—H···O, C—H···N, N—H···N and C—H···O (Table 2). The carbonyl O atoms of Gly¹ (O3) and Gly³ (O5) take part in the N4—H4D···O3 and N6—H6C···O5 hydrogen bonds, respectively, as shown in Fig. 1. The π electrons of the aromatic ring of Δ^EPhe⁴ take part in C—H···π interactions with atom H4B of Boc and atom H35A of L-Phe⁵. The distances and angles between C—H in alkyl group and the centre of the Δ^EPhe⁴ ring (denoted Cg1) are 3.883 Å and 150° for the C4—H4B···Cg1 contact and 3.557 Å and 172° for the C35—H35A···Cg1 contact, respectively (Table 2).

These results show that the presence of two α,β-dehydrophenylalanyl residues induces β turns in the pentapeptide Boc⁰—Gly¹–Δ^ZPhe²—Gly³–Δ^EPhe⁴—L-Phe⁵–p-NA. This is consistent with the results for other short peptides, such as the tetrapeptide Boc⁰—Gly¹–Δ^ZPhe²—Gly³—Phe⁴–p-NA (Ejsmont et al., 2001).

Table 2. Hydrogen-bonding geometry (Å, °). The C—H···π interaction is a hydrogen bond occurring between C—H in an alkyl group and the π system of Δ^EPhe⁴. The centroid of the Δ^EPhe⁴ ring is denoted Cg1.

S2. Experimental

The synthesis of Boc-Gly Δ^ZPhe has been described by Makowski et al. (1985). Boc-Gly Δ^EPhe-L-Phe-p-NA was obtained in the same manner as its Z isomer (Makowski et al., 2001); instead of TBTU [2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate], isobutyl chloroformate (0.26 ml, 2 mmol) in tetrahydrofuran (3.5 ml) was used. Isomers E and Z of Boc-Gly ΔPhe-L-Phe-p-NA were separated on a silica gel H-60 (Merck) column eluted with EtOAc (1–40%) in benzene. Yields of isomers E and Z were 18% and 53%, respectively. Gly Δ^EPhe-L-Phe-p-NA was obtained according to the method described by Makowski et al. (2001) and used for further synthesis without characterization. The only modification was that, instead of dissolution in ethyl ether and evaporation, the oily deblocked peptide was dissolved in propan-2-ol (20 ml) and precipitated with hexane. TFA (trifluoroacetic acid) (0.139 ml, 1 mmol) was added to a solution of Boc-Gly–Δ^ZPhe (0.16 g, 0.5 mmol) in tetrahydrofuran (2 ml) and the solution was cooled to 263 K. Isobutyl chloroformate (0.066 ml, 0.5 mmol) was then added and the mixture was left for 1.5 min at this temperature. Finally, Gly Δ^EPhe-L-Phe-p-NA (0.3 g, 0.5 mmol) was added and the reaction was carried out for 22 h at room temperature. The precipitate which formed was filtered off and the solvent was removed under reduced pressure. The resulting oil was dissolved in EtOAc (50 ml) and washed successively with 2 M HCl (2 × 3 ml), saturated potassium bicarbonate (3 × 3 ml) and brine (3 ml). The organic layer was dried over MgSO₄, the drying agent was removed by filtration and the solvent was evaporated. The product was crystallized from EtOAc–benzene (1:1)/hexane. The purity of the peptide (100%) was checked by high-perfomance liquid chromatography, using an Alltech Alltima column (C-18, 5 µm, 150 × 4.6 mm). Solvent system: A 0.1% TFA, B ACN (acetonitrile?), A:B 35:65, flow rate 1 ml min⁻¹. Yield 0.305 g (77%), m.p. 476–478 K. Elemental analysis, calculated for C₄₂H₄₃N₇O₉ (789.844): C 63.87, H 5.49%; found: C 64.04, H 5.28%. Long thin needle crystals of Boc⁰—Gly¹-Δ^ZPhe²—Gly³-Δ^EPhe⁴—L-Phe⁵-p-NA·C₂H₅OH, (I), suitable for X-ray structure analysis were grown at room temperature from a solution in ethanol. The crystals are sensitive and decompose in air.

S3. Refinement

All H atoms were placed in calculated positions, with C—H distances in the range 0.95–1.00 Å and N—H distances of 0.88 Å, and were allowed to ride on their parent atoms, with U_iso(H) = 1.2U_eq(C,N). Please check added text. The absolute structure was chosen on the basis of the known absolute configuration of the L-phenylalanine residue. The Friedel pairs were merged. Owing to the large anisotropic displacement parameter, the ethanol molecule is certainly slightly disordered but the type of disorder could not be resolved.

Computing details

Data collection: CrysAlis CCD (Oxford Diffraction, 2003); cell refinement: CrysAlis RED (Oxford Diffraction, 2003); data reduction: CrysAlis RED; program(s) used to solve structure: SHELXD (Sheldrick, 2002); program(s) used to refine structure: SHELXL97 (Sheldrick, 1997); molecular graphics: SHELXTL (Bruker, 1997); software used to prepare material for publication: SHELXL97.

Figures

Fig. 1. The molecular structure of (I), showing the atom-numbering scheme. Displacement ellipsoids are drawn at the ??% probability level and H atoms are shown as small spheres of arbitrary radii. Please provide missing information. Dashed lines indicate intramolecular hydrogen bonds.

N-[tert-Butoxycarbonylglycyl-(Z)-α,β-dehydrophenylalanylglycyl-(E)- α,β-dehydrophenylalanylphenylalanyl]-4-nitroaniline ethanol solvate

Crystal data

C42H43N7O9·C2H6O	F(000) = 884
Mr = 835.90	Dx = 1.299 Mg m^−3
Monoclinic, P21	Cu Kα radiation, λ = 1.5418 Å
Hall symbol: P 2yb	Cell parameters from 7064 reflections
a = 13.080 (4) Å	θ = 3–73°
b = 8.998 (3) Å	µ = 0.77 mm^−1
c = 18.406 (5) Å	T = 100 K
β = 99.37 (3)°	Long thin needle, yellow
V = 2137.4 (11) Å^3	0.45 × 0.04 × 0.02 mm
Z = 2

Data collection

Oxford Xcalibur PX κ-geometry diffractometer with CCD area detector	4423 independent reflections
Radiation source: fine-focus sealed tube	2794 reflections with I > 2σ(I)
Graphite monochromator	Rint = 0.085
ω and ϕ scans	θmax = 73.5°, θmin = 3.9°
Absorption correction: analytical CrysAlis RED (Oxford Diffraction, 2003)	h = −16→13
Tmin = 0.828, Tmax = 0.988	k = −9→11
14511 measured reflections	l = −22→19

Refinement

Refinement on F^2	Secondary atom site location: difference Fourier map
Least-squares matrix: full	Hydrogen site location: inferred from neighbouring sites
R[F^2 > 2σ(F^2)] = 0.072	H-atom parameters constrained
wR(F^2) = 0.192	w = 1/[σ^2(Fo^2) + (0.085P)^2] where P = (Fo^2 + 2Fc^2)/3
S = 1.10	(Δ/σ)max < 0.001
4423 reflections	Δρmax = 0.57 e Å^−3
552 parameters	Δρmin = −0.32 e Å^−3
1 restraint	Extinction correction: SHELXL97 (Sheldrick, 1997), Fc^*=kFc[1+0.001xFc^2λ^3/sin(2θ)]^-1/4
Primary atom site location: structure-invariant direct methods	Extinction coefficient: 0.0051 (7)

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.

Refinement. Refinement of F² against ALL reflections. The weighted R-factor wR and goodness of fit S are based on F², conventional R-factors R are based on F, with F set to zero for negative F². The threshold expression of F² > σ(F²) is used only for calculating R-factors(gt) etc. and is not relevant to the choice of reflections for refinement. R-factors based on F² are statistically about twice as large as those based on F, and R- factors based on ALL data will be even larger.

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

	x	y	z	Uiso*/Ueq
C1	0.0130 (5)	0.3239 (9)	0.9257 (5)	0.049 (2)
C2	−0.0488 (6)	0.4123 (11)	0.8636 (5)	0.057 (2)
H2A	−0.0724	0.5053	0.8831	0.068*
H2B	−0.1089	0.3540	0.8409	0.068*
H2C	−0.0050	0.4348	0.8265	0.068*
C3	−0.0471 (6)	0.2995 (12)	0.9868 (6)	0.066 (3)
H3A	−0.0653	0.3957	1.0062	0.079*
H3B	−0.0050	0.2425	1.0262	0.079*
H3C	−0.1105	0.2441	0.9683	0.079*
C4	0.0535 (7)	0.1804 (10)	0.8964 (6)	0.068 (3)
H4A	0.0911	0.1229	0.9375	0.081*
H4B	0.1002	0.2045	0.8615	0.081*
H4C	−0.0049	0.1217	0.8714	0.081*
O1	0.1017 (3)	0.4236 (5)	0.9486 (3)	0.0401 (12)
C5	0.1721 (5)	0.3895 (7)	1.0076 (4)	0.0343 (15)
O2	0.1736 (4)	0.2795 (6)	1.0468 (3)	0.0461 (13)
N1	0.2432 (4)	0.5021 (7)	1.0207 (4)	0.0420 (15)
H1D	0.2345	0.5819	0.9927	0.050*
C6	0.3318 (5)	0.4949 (9)	1.0784 (4)	0.0387 (16)
H6A	0.3127	0.4393	1.1207	0.046*
H6B	0.3514	0.5970	1.0954	0.046*
C7	0.4232 (5)	0.4213 (7)	1.0541 (3)	0.0276 (13)
O3	0.4193 (3)	0.3411 (5)	0.9989 (3)	0.0334 (10)
N2	0.5136 (4)	0.4473 (6)	1.1016 (3)	0.0295 (12)
H2D	0.5142	0.5120	1.1375	0.035*
C8	0.6062 (5)	0.3713 (7)	1.0937 (3)	0.0259 (13)
C9	0.6604 (5)	0.2886 (8)	1.1474 (4)	0.0304 (14)
H9A	0.7246	0.2525	1.1368	0.037*
C10	0.6370 (5)	0.2449 (7)	1.2194 (4)	0.0315 (14)
C11	0.5348 (5)	0.2412 (8)	1.2368 (4)	0.0370 (16)
H11A	0.4778	0.2728	1.2014	0.044*
C12	0.5184 (6)	0.1917 (9)	1.3051 (4)	0.0422 (17)
H12A	0.4500	0.1864	1.3161	0.051*
C13	0.6018 (6)	0.1497 (9)	1.3575 (4)	0.0465 (18)
H13A	0.5901	0.1173	1.4045	0.056*
C14	0.7019 (6)	0.1543 (8)	1.3422 (4)	0.0446 (18)
H14A	0.7585	0.1272	1.3790	0.053*
C15	0.7197 (5)	0.1987 (8)	1.2728 (4)	0.0358 (15)
H15A	0.7881	0.1977	1.2617	0.043*
C16	0.6463 (5)	0.3885 (7)	1.0232 (4)	0.0299 (14)
O4	0.7000 (4)	0.2891 (6)	1.0008 (3)	0.0469 (13)
N3	0.6195 (4)	0.5096 (6)	0.9815 (3)	0.0291 (12)
H3D	0.5812	0.5793	0.9971	0.035*
C17	0.6542 (5)	0.5249 (8)	0.9105 (4)	0.0333 (15)
H17A	0.7264	0.4884	0.9154	0.040*
H17B	0.6545	0.6317	0.8977	0.040*
C18	0.5882 (5)	0.4418 (7)	0.8476 (4)	0.0309 (14)
O5	0.6208 (3)	0.4256 (6)	0.7890 (2)	0.0343 (10)
N4	0.4946 (4)	0.3907 (6)	0.8597 (3)	0.0295 (12)
H4D	0.4777	0.4004	0.9038	0.035*
C19	0.4231 (5)	0.3222 (7)	0.8030 (4)	0.0279 (13)
C20	0.3204 (5)	0.3393 (8)	0.8024 (4)	0.0328 (15)
H20A	0.3004	0.4075	0.8369	0.039*
C21	0.2354 (5)	0.2618 (7)	0.7531 (4)	0.0312 (14)
C22	0.2420 (5)	0.1139 (7)	0.7331 (4)	0.0322 (15)
H22A	0.3038	0.0599	0.7499	0.039*
C23	0.1603 (5)	0.0435 (8)	0.6893 (4)	0.0392 (17)
H23A	0.1658	−0.0580	0.6762	0.047*
C24	0.0710 (5)	0.1222 (8)	0.6648 (4)	0.0394 (17)
H24A	0.0149	0.0744	0.6344	0.047*
C25	0.0617 (5)	0.2686 (8)	0.6835 (4)	0.0401 (17)
H25A	−0.0003	0.3217	0.6663	0.048*
C26	0.1435 (5)	0.3383 (8)	0.7278 (4)	0.0357 (16)
H26A	0.1371	0.4394	0.7412	0.043*
C27	0.4695 (5)	0.2233 (7)	0.7526 (4)	0.0277 (13)
O6	0.5207 (3)	0.1116 (5)	0.7765 (3)	0.0301 (10)
N5	0.4520 (4)	0.2578 (5)	0.6804 (3)	0.0283 (11)
H5A	0.4175	0.3397	0.6666	0.034*
C28	0.4876 (5)	0.1657 (7)	0.6239 (3)	0.0269 (13)
H28A	0.5180	0.0745	0.6499	0.032*
C29	0.3986 (5)	0.1114 (7)	0.5656 (4)	0.0301 (14)
H29A	0.4276	0.0471	0.5303	0.036*
H29B	0.3517	0.0494	0.5901	0.036*
C30	0.3360 (5)	0.2317 (7)	0.5233 (4)	0.0305 (14)
C31	0.3610 (5)	0.2834 (9)	0.4570 (4)	0.0417 (17)
H31A	0.4201	0.2437	0.4399	0.050*
C32	0.3033 (6)	0.3893 (10)	0.4157 (5)	0.054 (2)
H32A	0.3211	0.4193	0.3698	0.064*
C33	0.2190 (6)	0.4529 (9)	0.4406 (5)	0.053 (2)
H33A	0.1800	0.5289	0.4129	0.064*
C34	0.1924 (6)	0.4046 (9)	0.5061 (4)	0.0450 (19)
H34A	0.1347	0.4474	0.5239	0.054*
C35	0.2498 (5)	0.2937 (8)	0.5461 (4)	0.0386 (16)
H35A	0.2293	0.2593	0.5904	0.046*
C36	0.5741 (5)	0.2364 (7)	0.5911 (4)	0.0295 (13)
O7	0.5864 (3)	0.2075 (6)	0.5281 (3)	0.0391 (12)
N6	0.6379 (4)	0.3314 (6)	0.6366 (3)	0.0278 (11)
H6C	0.6208	0.3481	0.6802	0.033*
C37	0.7281 (5)	0.4057 (7)	0.6213 (4)	0.0287 (13)
C38	0.7667 (5)	0.3841 (7)	0.5549 (4)	0.0302 (14)
H38A	0.7362	0.3137	0.5194	0.036*
C39	0.8513 (5)	0.4699 (7)	0.5433 (4)	0.0335 (15)
H39A	0.8780	0.4597	0.4986	0.040*
C40	0.8962 (5)	0.5682 (8)	0.5953 (4)	0.0320 (14)
C41	0.8597 (5)	0.5863 (8)	0.6616 (4)	0.0356 (15)
H41A	0.8924	0.6538	0.6978	0.043*
C42	0.7747 (5)	0.5037 (8)	0.6736 (4)	0.0329 (15)
H42A	0.7484	0.5151	0.7184	0.039*
N7	0.9832 (4)	0.6591 (6)	0.5827 (4)	0.0367 (14)
O8	1.0103 (4)	0.6543 (6)	0.5212 (3)	0.0459 (14)
O9	1.0280 (4)	0.7377 (7)	0.6329 (4)	0.0580 (16)
O10	0.1999 (6)	0.2949 (8)	1.2030 (4)	0.085 (2)
H10A	0.1910	0.2909	1.1568	0.127*
C43	0.1370 (10)	0.4094 (13)	1.2255 (8)	0.101 (5)
H43A	0.1192	0.3845	1.2743	0.121*
H43C	0.0716	0.4163	1.1899	0.121*
C44	0.1953 (9)	0.5664 (13)	1.2301 (7)	0.086 (4)
H44A	0.2012	0.6001	1.1803	0.129*
H44B	0.2647	0.5556	1.2591	0.129*
H44D	0.1558	0.6394	1.2536	0.129*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
C1	0.026 (3)	0.044 (4)	0.073 (6)	−0.008 (3)	0.000 (4)	−0.004 (4)
C2	0.043 (4)	0.055 (5)	0.066 (6)	−0.002 (4)	−0.012 (4)	0.011 (5)
C3	0.037 (4)	0.067 (6)	0.095 (8)	−0.004 (4)	0.016 (5)	0.021 (6)
C4	0.062 (5)	0.037 (5)	0.097 (8)	0.005 (4)	−0.009 (5)	−0.012 (5)
O1	0.031 (2)	0.032 (3)	0.055 (3)	−0.004 (2)	−0.002 (2)	0.009 (3)
C5	0.028 (3)	0.028 (3)	0.043 (4)	−0.004 (3)	−0.003 (3)	−0.001 (3)
O2	0.047 (3)	0.032 (3)	0.058 (4)	0.002 (2)	0.005 (3)	0.015 (3)
N1	0.038 (3)	0.031 (3)	0.053 (4)	0.002 (3)	−0.003 (3)	0.007 (3)
C6	0.031 (3)	0.044 (4)	0.040 (4)	0.007 (3)	0.003 (3)	−0.005 (3)
C7	0.033 (3)	0.024 (3)	0.025 (3)	−0.005 (3)	0.002 (2)	−0.002 (3)
O3	0.042 (2)	0.031 (2)	0.027 (2)	−0.006 (2)	0.004 (2)	−0.003 (2)
N2	0.031 (3)	0.031 (3)	0.025 (3)	0.004 (2)	0.002 (2)	−0.007 (2)
C8	0.031 (3)	0.026 (3)	0.021 (3)	0.003 (2)	0.004 (2)	−0.003 (3)
C9	0.032 (3)	0.033 (3)	0.027 (3)	0.002 (3)	0.007 (3)	0.002 (3)
C10	0.044 (3)	0.026 (3)	0.026 (3)	0.002 (3)	0.009 (3)	−0.003 (3)
C11	0.045 (4)	0.036 (4)	0.032 (4)	0.000 (3)	0.010 (3)	0.000 (3)
C12	0.054 (4)	0.041 (4)	0.035 (4)	−0.006 (3)	0.018 (3)	−0.004 (3)
C13	0.065 (5)	0.038 (4)	0.034 (4)	−0.009 (4)	0.001 (4)	−0.003 (3)
C14	0.057 (5)	0.030 (4)	0.044 (4)	−0.001 (3)	0.000 (4)	0.002 (3)
C15	0.042 (4)	0.028 (3)	0.035 (4)	0.008 (3)	−0.001 (3)	0.011 (3)
C16	0.034 (3)	0.031 (3)	0.026 (3)	0.001 (3)	0.007 (3)	−0.003 (3)
O4	0.060 (3)	0.039 (3)	0.048 (3)	0.018 (2)	0.027 (3)	0.007 (3)
N3	0.039 (3)	0.023 (3)	0.028 (3)	−0.002 (2)	0.013 (2)	−0.001 (2)
C17	0.036 (3)	0.038 (4)	0.027 (3)	−0.007 (3)	0.008 (3)	−0.006 (3)
C18	0.037 (3)	0.024 (3)	0.033 (4)	−0.004 (3)	0.008 (3)	0.000 (3)
O5	0.038 (2)	0.039 (3)	0.028 (2)	−0.007 (2)	0.012 (2)	−0.002 (2)
N4	0.025 (2)	0.032 (3)	0.031 (3)	−0.010 (2)	0.006 (2)	−0.004 (2)
C19	0.031 (3)	0.031 (3)	0.024 (3)	−0.002 (3)	0.009 (3)	−0.001 (3)
C20	0.031 (3)	0.032 (3)	0.035 (4)	0.001 (3)	0.004 (3)	0.000 (3)
C21	0.028 (3)	0.025 (3)	0.040 (4)	−0.002 (2)	0.004 (3)	−0.002 (3)
C22	0.032 (3)	0.025 (3)	0.040 (4)	−0.001 (3)	0.005 (3)	0.003 (3)
C23	0.038 (4)	0.030 (4)	0.049 (5)	−0.001 (3)	0.005 (3)	−0.005 (3)
C24	0.032 (3)	0.036 (4)	0.048 (4)	−0.003 (3)	−0.001 (3)	0.000 (3)
C25	0.029 (3)	0.039 (4)	0.052 (5)	−0.003 (3)	0.005 (3)	0.000 (4)
C26	0.032 (3)	0.026 (3)	0.047 (4)	0.007 (3)	0.000 (3)	−0.001 (3)
C27	0.031 (3)	0.018 (3)	0.035 (4)	−0.002 (2)	0.010 (3)	0.004 (3)
O6	0.031 (2)	0.025 (2)	0.033 (2)	0.0011 (18)	0.0034 (19)	0.006 (2)
N5	0.039 (3)	0.018 (2)	0.028 (3)	0.002 (2)	0.006 (2)	0.001 (2)
C28	0.030 (3)	0.025 (3)	0.027 (3)	−0.005 (2)	0.008 (3)	−0.011 (3)
C29	0.031 (3)	0.029 (3)	0.028 (3)	−0.001 (3)	−0.004 (3)	−0.006 (3)
C30	0.032 (3)	0.023 (3)	0.036 (3)	0.003 (2)	0.003 (3)	−0.002 (3)
C31	0.029 (3)	0.056 (5)	0.038 (4)	0.000 (3)	0.000 (3)	0.004 (4)
C32	0.039 (4)	0.061 (5)	0.056 (5)	−0.004 (4)	−0.007 (4)	0.022 (5)
C33	0.047 (4)	0.036 (4)	0.069 (6)	−0.005 (3)	−0.015 (4)	0.006 (4)
C34	0.043 (4)	0.034 (4)	0.051 (5)	0.009 (3)	−0.010 (3)	−0.015 (4)
C35	0.037 (3)	0.038 (4)	0.038 (4)	−0.002 (3)	0.000 (3)	−0.013 (3)
C36	0.030 (3)	0.029 (3)	0.029 (3)	−0.001 (3)	0.005 (3)	−0.001 (3)
O7	0.039 (2)	0.053 (3)	0.027 (3)	−0.001 (2)	0.010 (2)	−0.013 (2)
N6	0.029 (2)	0.027 (3)	0.028 (3)	−0.008 (2)	0.007 (2)	−0.003 (2)
C37	0.030 (3)	0.031 (3)	0.027 (3)	0.002 (3)	0.011 (3)	0.008 (3)
C38	0.035 (3)	0.028 (3)	0.028 (3)	−0.003 (3)	0.006 (3)	−0.001 (3)
C39	0.032 (3)	0.034 (4)	0.036 (4)	0.003 (3)	0.009 (3)	0.009 (3)
C40	0.026 (3)	0.029 (3)	0.042 (4)	0.003 (3)	0.010 (3)	0.005 (3)
C41	0.032 (3)	0.032 (3)	0.042 (4)	0.001 (3)	0.005 (3)	−0.001 (3)
C42	0.034 (3)	0.036 (4)	0.029 (3)	−0.010 (3)	0.007 (3)	−0.005 (3)
N7	0.031 (3)	0.031 (3)	0.050 (4)	−0.004 (2)	0.012 (3)	0.005 (3)
O8	0.038 (3)	0.046 (3)	0.058 (4)	0.003 (2)	0.021 (3)	0.012 (3)
O9	0.046 (3)	0.059 (4)	0.073 (4)	−0.019 (3)	0.020 (3)	−0.013 (3)
O10	0.114 (6)	0.070 (5)	0.075 (5)	0.030 (5)	0.026 (5)	0.026 (4)
C43	0.117 (9)	0.073 (8)	0.127 (11)	0.045 (7)	0.062 (9)	0.061 (8)
C44	0.093 (8)	0.070 (7)	0.097 (9)	0.001 (6)	0.023 (7)	0.016 (7)

Geometric parameters (Å, º)

C1—O1	1.472 (8)	C20—H20A	0.9500
C1—C3	1.489 (12)	C21—C22	1.387 (9)
C1—C2	1.514 (11)	C21—C26	1.397 (9)
C1—C4	1.528 (12)	C22—C23	1.383 (9)
C2—H2A	0.9800	C22—H22A	0.9500
C2—H2B	0.9800	C23—C24	1.379 (10)
C2—H2C	0.9800	C23—H23A	0.9500
C3—H3A	0.9800	C24—C25	1.372 (10)
C3—H3B	0.9800	C24—H24A	0.9500
C3—H3C	0.9800	C25—C26	1.385 (10)
C4—H4A	0.9800	C25—H25A	0.9500
C4—H4B	0.9800	C26—H26A	0.9500
C4—H4C	0.9800	C27—O6	1.247 (7)
O1—C5	1.340 (8)	C27—N5	1.348 (8)
C5—O2	1.223 (8)	N5—H5A	0.8800
C5—N1	1.370 (9)	C28—C36	1.507 (8)
N1—C6	1.441 (9)	C28—H28A	1.0000
N1—H1D	0.8800	C29—C30	1.497 (9)
C6—C7	1.497 (9)	C29—H29A	0.9900
C6—H6A	0.9900	C29—H29B	0.9900
C6—H6B	0.9900	C30—C35	1.383 (9)
C7—O3	1.241 (7)	C30—C31	1.394 (10)
C7—N2	1.371 (7)	C31—C32	1.367 (11)
N2—C8	1.419 (7)	C31—H31A	0.9500
N4—C19	1.423 (8)	C32—C33	1.386 (12)
N5—C28	1.464 (7)	C32—H32A	0.9500
N2—H2D	0.8800	C33—C34	1.379 (12)
C8—C9	1.344 (9)	C33—H33A	0.9500
C19—C20	1.351 (8)	C34—C35	1.387 (10)
C28—C29	1.529 (8)	C34—H34A	0.9500
C8—C16	1.484 (9)	C35—H35A	0.9500
C9—C10	1.462 (9)	C36—O7	1.225 (8)
C9—H9A	0.9500	C36—N6	1.378 (8)
C10—C15	1.400 (9)	N6—C37	1.424 (8)
C10—C11	1.425 (9)	N6—H6C	0.8800
C11—C12	1.383 (10)	C37—C42	1.373 (9)
C11—H11A	0.9500	C37—C38	1.410 (9)
C12—C13	1.386 (11)	C38—C39	1.393 (9)
C12—H12A	0.9500	C38—H38A	0.9500
C13—C14	1.384 (11)	C39—C40	1.363 (10)
C13—H13A	0.9500	C39—H39A	0.9500
C14—C15	1.394 (10)	C40—C41	1.391 (10)
C14—H14A	0.9500	C40—N7	1.450 (8)
C15—H15A	0.9500	C41—C42	1.385 (9)
C16—O4	1.248 (8)	C41—H41A	0.9500
C16—N3	1.345 (8)	C42—H42A	0.9500
N3—C17	1.457 (8)	N7—O9	1.234 (8)
N3—H3D	0.8800	N7—O8	1.242 (8)
C17—C18	1.523 (9)	O10—C43	1.422 (13)
C17—H17A	0.9900	O10—H10A	0.8400
C17—H17B	0.9900	C43—C44	1.601 (17)
C18—O5	1.230 (8)	C43—H43A	0.9900
C18—N4	1.360 (8)	C43—H43C	0.9900
N4—H4D	0.8800	C44—H44A	0.9800
C19—C27	1.485 (8)	C44—H44B	0.9800
C20—C21	1.490 (9)	C44—H44D	0.9800
O1—C1—C3	111.2 (7)	C26—C21—C20	119.2 (6)
O1—C1—C2	101.3 (6)	C23—C22—C21	121.2 (6)
C3—C1—C2	111.4 (7)	C23—C22—H22A	119.4
O1—C1—C4	108.3 (6)	C21—C22—H22A	119.4
C3—C1—C4	113.4 (8)	C24—C23—C22	119.3 (7)
C2—C1—C4	110.5 (8)	C24—C23—H23A	120.4
C1—C2—H2A	109.5	C22—C23—H23A	120.4
C1—C2—H2B	109.5	C25—C24—C23	121.0 (7)
H2A—C2—H2B	109.5	C25—C24—H24A	119.5
C1—C2—H2C	109.5	C23—C24—H24A	119.5
H2A—C2—H2C	109.5	C24—C25—C26	119.4 (7)
H2B—C2—H2C	109.5	C24—C25—H25A	120.3
C1—C3—H3A	109.5	C26—C25—H25A	120.3
C1—C3—H3B	109.5	C25—C26—C21	120.8 (7)
H3A—C3—H3B	109.5	C25—C26—H26A	119.6
C1—C3—H3C	109.5	C21—C26—H26A	119.6
H3A—C3—H3C	109.5	O6—C27—N5	121.9 (6)
H3B—C3—H3C	109.5	O6—C27—C19	120.9 (6)
C1—C4—H4A	109.5	N5—C27—C19	117.2 (5)
C1—C4—H4B	109.5	C27—N5—C28	123.2 (5)
H4A—C4—H4B	109.5	C27—N5—H5A	118.4
C1—C4—H4C	109.5	C28—N5—H5A	118.4
H4A—C4—H4C	109.5	N5—C28—C36	113.1 (5)
H4B—C4—H4C	109.5	N5—C28—C29	112.7 (5)
C5—O1—C1	119.9 (6)	C36—C28—C29	112.9 (5)
O2—C5—O1	127.3 (6)	N5—C28—H28A	105.8
O2—C5—N1	123.4 (6)	C36—C28—H28A	105.8
O1—C5—N1	109.3 (6)	C29—C28—H28A	105.8
C5—N1—C6	122.5 (6)	C30—C29—C28	115.1 (5)
C5—N1—H1D	118.7	C30—C29—H29A	108.5
C6—N1—H1D	118.7	C28—C29—H29A	108.5
N1—C6—C7	112.6 (6)	C30—C29—H29B	108.5
N1—C6—H6A	109.1	C28—C29—H29B	108.5
C7—C6—H6A	109.1	H29A—C29—H29B	107.5
N1—C6—H6B	109.1	C35—C30—C31	116.8 (6)
C7—C6—H6B	109.1	C35—C30—C29	122.5 (7)
H6A—C6—H6B	107.8	C31—C30—C29	120.7 (6)
O3—C7—N2	122.4 (6)	C32—C31—C30	122.2 (7)
O3—C7—C6	125.0 (6)	C32—C31—H31A	118.9
N2—C7—C6	112.5 (5)	C30—C31—H31A	118.9
C7—N2—C8	121.0 (5)	C31—C32—C33	120.1 (8)
C7—N2—H2D	119.5	C31—C32—H32A	120.0
C8—N2—H2D	119.5	C33—C32—H32A	120.0
C9—C8—N2	123.2 (6)	C34—C33—C32	119.2 (8)
C9—C8—C16	118.8 (6)	C34—C33—H33A	120.4
N2—C8—C16	117.9 (5)	C32—C33—H33A	120.4
C8—C9—C10	130.8 (6)	C33—C34—C35	120.0 (7)
C8—C9—H9A	114.6	C33—C34—H34A	120.0
C10—C9—H9A	114.6	C35—C34—H34A	120.0
C15—C10—C11	118.8 (6)	C30—C35—C34	121.8 (7)
C15—C10—C9	117.7 (6)	C30—C35—H35A	119.1
C11—C10—C9	123.5 (6)	C34—C35—H35A	119.1
C12—C11—C10	120.1 (7)	O7—C36—N6	123.1 (6)
C12—C11—H11A	120.0	O7—C36—C28	120.8 (6)
C10—C11—H11A	120.0	N6—C36—C28	116.0 (5)
C11—C12—C13	120.0 (7)	C36—N6—C37	127.6 (5)
C11—C12—H12A	120.0	C36—N6—H6C	116.2
C13—C12—H12A	120.0	C37—N6—H6C	116.2
C14—C13—C12	121.0 (8)	C42—C37—C38	121.0 (6)
C14—C13—H13A	119.5	C42—C37—N6	116.8 (5)
C12—C13—H13A	119.5	C38—C37—N6	122.1 (6)
C13—C14—C15	119.9 (7)	C39—C38—C37	117.5 (6)
C13—C14—H14A	120.0	C39—C38—H38A	121.3
C15—C14—H14A	120.0	C37—C38—H38A	121.3
C14—C15—C10	120.2 (7)	C40—C39—C38	121.0 (6)
C14—C15—H15A	119.9	C40—C39—H39A	119.5
C10—C15—H15A	119.9	C38—C39—H39A	119.5
O4—C16—N3	120.0 (6)	C39—C40—C41	121.4 (6)
O4—C16—C8	120.8 (6)	C39—C40—N7	121.0 (6)
N3—C16—C8	119.1 (5)	C41—C40—N7	117.6 (6)
C16—N3—C17	119.8 (5)	C42—C41—C40	118.5 (7)
C16—N3—H3D	120.1	C42—C41—H41A	120.8
C17—N3—H3D	120.1	C40—C41—H41A	120.8
N3—C17—C18	114.5 (5)	C37—C42—C41	120.6 (6)
N3—C17—H17A	108.6	C37—C42—H42A	119.7
C18—C17—H17A	108.6	C41—C42—H42A	119.7
N3—C17—H17B	108.6	O9—N7—O8	122.1 (6)
C18—C17—H17B	108.6	O9—N7—C40	119.6 (6)
H17A—C17—H17B	107.6	O8—N7—C40	118.3 (6)
O5—C18—N4	123.6 (6)	C43—O10—H10A	109.5
O5—C18—C17	119.4 (6)	O10—C43—C44	111.1 (9)
N4—C18—C17	117.1 (6)	O10—C43—H43A	109.4
C18—N4—C19	121.9 (6)	C44—C43—H43A	109.4
C18—N4—H4D	119.0	O10—C43—H43C	109.4
C19—N4—H4D	119.0	C44—C43—H43C	109.4
C20—C19—N4	119.4 (6)	H43A—C43—H43C	108.0
C20—C19—C27	124.8 (6)	C43—C44—H44A	109.5
N4—C19—C27	115.5 (5)	C43—C44—H44B	109.5
C19—C20—C21	126.5 (6)	H44A—C44—H44B	109.5
C19—C20—H20A	116.8	C43—C44—H44D	109.5
C21—C20—H20A	116.8	H44A—C44—H44D	109.5
C22—C21—C26	118.2 (6)	H44B—C44—H44D	109.5
C22—C21—C20	122.6 (6)
C3—C1—O1—C5	56.3 (9)	C23—C24—C25—C26	0.0 (12)
C2—C1—O1—C5	174.8 (7)	C24—C25—C26—C21	0.5 (11)
C4—C1—O1—C5	−69.0 (9)	C22—C21—C26—C25	−0.6 (11)
C1—O1—C5—O2	1.1 (11)	C20—C21—C26—C25	−178.2 (7)
O1—C5—N1—C6	−176.9 (6)	C20—C19—C27—O6	−113.2 (8)
C1—O1—C5—N1	−176.4 (6)	N4—C19—C27—O6	60.5 (8)
O2—C5—N1—C6	5.5 (11)	C20—C19—C27—N5	65.4 (9)
C5—N1—C6—C7	87.5 (8)	N4—C19—C27—N5	−120.8 (6)
N1—C6—C7—O3	−17.8 (10)	O6—C27—N5—C28	2.4 (9)
N1—C6—C7—N2	164.5 (6)	C19—C27—N5—C28	−176.3 (5)
O3—C7—N2—C8	−6.3 (9)	C27—N5—C28—C36	−109.7 (7)
C6—C7—N2—C8	171.5 (6)	C27—N5—C28—C29	120.7 (6)
C7—N2—C8—C9	−122.8 (7)	N5—C28—C29—C30	60.4 (7)
C7—N2—C8—C16	59.1 (8)	C36—C28—C29—C30	−69.3 (7)
N2—C8—C9—C10	6.3 (11)	C28—C29—C30—C35	−89.6 (8)
C16—C8—C9—C10	−175.6 (7)	C28—C29—C30—C31	92.3 (8)
C8—C9—C10—C15	−160.4 (7)	C35—C30—C31—C32	−0.5 (11)
C8—C9—C10—C11	22.3 (11)	C29—C30—C31—C32	177.6 (7)
C15—C10—C11—C12	−0.5 (10)	C30—C31—C32—C33	2.4 (13)
C9—C10—C11—C12	176.8 (7)	C31—C32—C33—C34	−2.0 (12)
C10—C11—C12—C13	1.9 (11)	C32—C33—C34—C35	0.0 (12)
C11—C12—C13—C14	−1.0 (12)	C31—C30—C35—C34	−1.6 (10)
C12—C13—C14—C15	−1.3 (12)	C29—C30—C35—C34	−179.8 (6)
C13—C14—C15—C10	2.8 (11)	C33—C34—C35—C30	1.9 (11)
C11—C10—C15—C14	−1.9 (10)	N5—C28—C36—O7	−152.2 (6)
C9—C10—C15—C14	−179.3 (7)	C29—C28—C36—O7	−22.7 (9)
C9—C8—C16—O4	29.1 (9)	N5—C28—C36—N6	28.9 (8)
N2—C8—C16—O4	−152.8 (6)	C29—C28—C36—N6	158.4 (5)
C9—C8—C16—N3	−154.5 (6)	O7—C36—N6—C37	−2.7 (10)
N2—C8—C16—N3	23.7 (9)	C28—C36—N6—C37	176.1 (6)
O4—C16—N3—C17	−0.6 (9)	C36—N6—C37—C42	175.4 (6)
C8—C16—N3—C17	−177.1 (6)	C36—N6—C37—C38	−2.7 (10)
C16—N3—C17—C18	82.2 (8)	C42—C37—C38—C39	−2.3 (9)
N3—C17—C18—O5	−167.2 (6)	N6—C37—C38—C39	175.7 (6)
N3—C17—C18—N4	13.6 (9)	C37—C38—C39—C40	1.6 (9)
O5—C18—N4—C19	−4.2 (10)	C38—C39—C40—C41	0.1 (10)
C17—C18—N4—C19	174.9 (6)	C38—C39—C40—N7	−179.0 (6)
C18—N4—C19—C20	−147.2 (7)	C39—C40—C41—C42	−1.1 (10)
C18—N4—C19—C27	38.6 (8)	N7—C40—C41—C42	178.0 (6)
N4—C19—C20—C21	−172.3 (6)	C38—C37—C42—C41	1.4 (10)
C27—C19—C20—C21	1.2 (11)	N6—C37—C42—C41	−176.7 (6)
C19—C20—C21—C22	38.3 (11)	C40—C41—C42—C37	0.4 (10)
C19—C20—C21—C26	−144.2 (7)	C39—C40—N7—O9	−173.8 (6)
C26—C21—C22—C23	0.2 (11)	C41—C40—N7—O9	7.1 (9)
C20—C21—C22—C23	177.7 (7)	C39—C40—N7—O8	5.8 (9)
C21—C22—C23—C24	0.3 (11)	C41—C40—N7—O8	−173.3 (6)
C22—C23—C24—C25	−0.4 (12)

Hydrogen-bond geometry (Å, º)

D—H···A	D—H	H···A	D···A	D—H···A
N1—H1D···O4^i	0.88	2.05	2.734 (8)	134
N2—H2D···O6^i	0.88	1.94	2.784 (7)	161
N3—H3D···O3^i	0.88	2.36	3.057 (7)	137
C6—H6B···O4^i	0.99	2.49	3.018 (10)	113
C13—H13A···O7^ii	0.95	2.42	3.222 (9)	142
C3—H3B···O2	0.98	2.33	2.924 (10)	118
C4—H4A···O2	0.98	2.55	3.085 (12)	114
C31—H31A···O7	0.95	2.51	3.101 (8)	120
C38—H38A···O7	0.95	2.21	2.820 (8)	121
C11—H11A···N2	0.95	2.52	3.080 (9)	118
N4—H4D···O3	0.88	2.09	2.925 (7)	158
N6—H6C···O5	0.88	2.12	2.974 (7)	163
N4—H4D···N3	0.88	2.36	2.766 (8)	108
N6—H6C···N5	0.88	2.35	2.764 (7)	109
O10—H10A···O2	0.84	2.00	2.843 (10)	178
C4—H4B···Cg1	0.98	3.00	3.883	150
C35—H35A···Cg1	0.95	2.61	3.557	172

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