The tetrapeptide Z-Leu-Aib-Pro-Val-OBg monohydrate

The intramolecular hydrogen-bonding pattern of Z-Leu-Aib-Pro-Val-OBg monohydrate [(N-benzhydryl­amino)­carbonyl­methyl N-benzyl­oxy­carbonyl-[alpha]-amino­isobutyryl­prolyl­valinate monohydrate], C₄₃H₅₅N₅O₈·H₂O, is unusual for a tetrapeptide because, in addition to a 14 hydrogen bond, a second hydrogen bond of the type 15 is formed. This folding reflects the intramolecular hydrogen-bonding pattern that this amino acid sequence adopts in the naturally occurring peptaibol alamethicin.

S1. Comment

The sequence Leu-Aib-Pro-Val represents a segment of the naturally occurring peptaibol antibiotics [peptides containing α-aminoisobutyric acid (Aib) and a C-terminal β-amino alcohol (Brückner & Graf, 1983; Benedetti et al., 1982)] such as alamethicin (Pandey et al., 1977; Brückner et al., 1985), gliodeliquescin (Brückner & Przybylski, 1984), trichobrachin (Kripp, 1990), trichocellin (Wada et al., 1994), trichokonin (Huang et al., 1995a & 1995b) and trichorzianin (Bodo et al., 1985). OBg is an unique protecting group for the C-termini of synthetic peptides, since it can be cleaved under very mild alkaline conditions, such as treatment with aqueous 10% sodium carbonate (Amblard & Rodriguez, 1988). The backbone conformation that the tetrapeptide adopts is quite unusual. Leu1 lies with its torsion angles (Table 2) in the semi-extended region of a Ramachandran plot, which is unusual for a N-terminal protected residue of an Aib-containing peptide. In most other cases, the C═O group of the protection group is involved in an intramolecular hydrogen bond with the third or fourth residue, thereby forcing the first residue to adopt a helical conformation (for examples see Geßmann et al., 1991, 1997; Geßmann, 1999). In (I), the third residue is proline, which cannot act as a hydrogen-bond donor, and instead the Val4 N atom forms a 1←4 hydrogen bond with Leu1. Furthermore, the ϕ/ψ values of Leu1 are in an allowed, albeit not the most favoured, region of the Ramachandran plot. While Aib2 and Pro3 are in the most favoured helical region, Val4 is again just in the additional allowed helical region. A larger variation of the torsion angles for the standard amino acids Val and Leu in Aib-containing peptides as compared with those in proteins has been observed previously (Geßmann et al., 1994 & 1997, Geßmann, 1999) and is probably the result of the smaller number of intramolecular hydrogen bonds in peptides.

There are two intramolecular hydrogen bonds in (I). One is the 1← 4 hydrogen bond between the Val4 NH group and the C═O group of Leu1, which forms a ring with the usual (for an incipient 3₁₀-helix) ten atoms. The second hydrogen bond is formed between the NH-group of OBg and the C═O group of Aib2. The number of ring atoms is 13, which is typical for an α-helical 1← 5 hydrogen bond. A hydrogen bond of that type has, to our knowledge, never been observed in a tetrapeptide and is clearly due to the substitution of the amino group in a residue for an O atom in the OBg protection group, prohibiting a 1←4 interaction between C═O (Aib2) and O (OBg). Interestingly, the only crystal structure of a natural peptide that comprises the sequence Val-Aib-Pro-Leu, that of alamethicin (Fox & Richards, 1982), shows a similar hydrogen-bond pattern, viz. no intramolecular hydrogen bond involving the C═O group of the residue preceeding Leu12, a 1←4 hydrogen bond between the C═O group of Leu12 and the N—H group of Val15, and a 1←5 hydrogen bond between the C═O group of Aib13 and the N—H group of residue 17, which is in the place of the N—H group of the OBg protection group in alamethicin. The peptide unit in (I) adopts the usual trans planar conformation with an average deviation of 6.1° from ideal geometry (ω = 180°), a deviation which is unusually large, especially for Leu1 (Table 2). The side chain of Leu1 adopts the most common conformation (Janin et al., 1978), which belongs to the type g⁺ for χ₁ and g⁺t for χ₂. The pyrrolidine ring of Pro3 adopts the C_γ-endo (Ashida & Kakudo, 1974) conformation [Cremer & Pople (1975) puckering parameters Q=0.368 Å and Φ=281.9°]. The angles between the N-terminal benzene plane and the two C-terminal benzene planes are 66.7 (4) and 79.6 (4)°, respectively, while the angle between the two C-terminal planes is 80.2 (4)°.

In the crystal packing, the unique molecule is hydrogen bonded to a symmetry-related molecule at (x − 1,y,z), thus building infinite long columns in the [−1 0 0] direction. Each molecule is also hydrogen bonded along [0 − 1 0] via the co-crystallized water molecule (Table 1 and Fig. 2). Thus, planes of hydrogen bonded molecules are formed parallel to the ab plane. These planes stack via apolar molecular contacts along the c axis.

S2. Experimental

Z-Leu-Aib-Pro-Val-OBg was synthesized in the following way: To Z-Leu-Aib-Pro-OH (225 mg, 0.5 mmol) in N,N-dimethylformamide (DMF; 10 ml) were added (benzotriazol-1-yloxy)tris(pyrrolidino)phosphonium hexafluorophosphate (PyBOP; 260 mg, 0.5 mmol; Novabiochem, Läufelfingen, Switzerland), H-Val-OBg x trifluoroacetic acid (TFA; 250 mg, 0.5 mmol; Fluka, Buchs, Switzerland) and N-methylmorpholine (NMM; 112 µl, 1.0 mmol; Fluka, Buchs, Switzerland). After 16 h at room temperature, the solvent was removed in vacuo. The remaining residue was dissolved in ethyl acetate (100 ml) and washed successively (3 x 50 ml) with KHSO₄ (5%), KHCO₃ (5%) and water. The organic phase was dried overnight over anhydrous Na₂SO₄ and evaporated to dryness. To the remaining oil, a small amount of methanol was added, and the peptide was precipitated by addition of diethyl ether and n-hexane. Yield 385 mg (73%); m.p. 368 K. Analysis calculated for C₄₃H₅₅N₅O₈·H₂O (787.94): C 65.83, H 7.29, N 8.89%; found: C 65.80, H. 7.21, N 8.75%. Enantioselective gas chromatography on Chirasil-Val of a total hydrolysate of the title compound revealed less than 0.7% racemization of L-amino acids. The tripeptide Z-Leu-Aib-Pro-OH was prepared by conventional stepwise solution phase synthesis, as described by Brückner & Koza (2003). H-Val-OBg x TFA was obtained by treatment of Boc-Val-OBg (Novabiochem, Läufelfingen, Switzerland) with TFA. The tetrapeptide was crystallized by cooling a hot (343 K) methanol–water mixture (70:30) to room temperature. Rod-shaped crystals suitable for X-ray analysis were obtained after some days.

S3. Refinement

Data were collected from θ=1–60°, using a collimator with a diameter of 1 mm, resulting in 6444 unique reflections. Refinement against these reflections resulted in an unreasonable Flack parameter of −0.6 (4). Therefore, 2743 Friedel pairs were merged before repeated refinement. All but five non-H atoms were detected by direct methods with the program Crisp, and the remaining non-H atoms were located from the difference Fourier syntheses. H atoms were placed at calculated postions and refined as riding. The highest peak in the final difference Fourier synthesis was associated with a water H atom, although this H atom has no acceptor. All U_eq(H) values were fixed to 1.5U_eq of the parent atom, except for H atoms belonging to the water molecule, where U_eq(H) was fixed to 1.2U_eq of the O atom. These H atoms were refined with distance constraints (0.96 Å) also restraining the H—O—H angle. The largest positive residual electron density is close (0.24 Å) to the water H atom without an acceptor.

Computing details

Data collection: CAD-4 (Enraf–Nonius, 1989); cell refinement: CAD-4; data reduction: Xtal3.7 DIFDAT, ABSORB, ADDREF (du Bouley & Hall, 2000); program(s) used to solve structure: CRISP (du Bouley & Hall, 2000); program(s) used to refine structure: SHELXL97 (Sheldrick, 1997); molecular graphics: Xtal3.7 PIG (du Bouley & Hall, 2000), Xtal3.7 ORTEP (Johnson, 1976, Davenport et al., 2000), Setor (Evans, 1993); software used to prepare material for publication: SHELXL97, Xtal3.7 CIFIO, BONDLA (du Bouley & Hall, 2000), PLATON (Spek, 2002).

Figures

	Fig. 1. The molecular structure of (I), showing displacement ellipsoids at the 30% probability level.
	Fig. 2. The crystal packing of (I), viewed approximately along [0 1 1]. The four hydrogen bonds, by translation of the unit cell symmetry- related molecules, are shown. Hydrogen bonds are indicated as broken lines and H atoms have been omitted for clarity.

(N-benzhydrylamino)carbonylmethyl N-benzyloxycarbonyl-α-aminoisobutyrylprolylvalinate monohydrate

Crystal data

C43H55N5O8·H2O	F(000) = 1688
Mr = 787.94	Dx = 1.191 Mg m^−3
Orthorhombic, P212121	Cu Kα radiation, λ = 1.54178 Å
Hall symbol: P 2ac 2ab	Cell parameters from 25 reflections
a = 9.882 (11) Å	θ = 13.5–18.9°
b = 10.705 (6) Å	µ = 0.68 mm^−1
c = 41.553 (17) Å	T = 293 K
V = 4396 (6) Å^3	Rod, colourless
Z = 4	0.8 × 0.1 × 0.1 mm

Data collection

Enraf–Nonius CAD-4 diffractometer	2471 reflections with I > 2σ(I)
Radiation source: fine-focus sealed tube	Rint = 0.110
Graphite monochromator	θmax = 59.9°, θmin = 2.1°
ω/θ scans	h = −11→11
Absorption correction: analytical ABSORB (Bouley & Hall, 2000)	k = −11→12
Tmin = 0.888, Tmax = 0.953	l = −46→46
7588 measured reflections	5 standard reflections every 120 min
3701 independent reflections	intensity decay: 2%

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)] = 0.061	H atoms treated by a mixture of independent and constrained refinement
wR(F^2) = 0.184	w = 1/[σ^2(Fo^2) + (0.0824P)^2] where P = (Fo^2 + 2Fc^2)/3
S = 1.08	(Δ/σ)max = 0.010
3701 reflections	Δρmax = 0.52 e Å^−3
521 parameters	Δρmin = −0.24 e Å^−3
3 restraints	Extinction correction: SHELXL97, Fc^*=kFc[1+0.001xFc^2λ^3/sin(2θ)]^-1/4
Primary atom site location: structure-invariant direct methods	Extinction coefficient: 0.00064 (15)

Special details

Geometry. All e.s.d.'s 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
C01	0.8669 (7)	−0.1220 (6)	0.55838 (14)	0.0622 (16)
C02	0.9952 (8)	−0.1057 (7)	0.57105 (18)	0.081 (2)
H02	1.0199	−0.1451	0.5901	0.122*
C03	1.0862 (8)	−0.0297 (9)	0.5548 (2)	0.098 (3)
H03	1.1713	−0.0170	0.5638	0.147*
C04	1.0572 (9)	0.0260 (8)	0.5269 (2)	0.089 (2)
H04	1.1219	0.0729	0.5160	0.134*
C05	0.9269 (9)	0.0117 (9)	0.51456 (18)	0.096 (3)
H05	0.9027	0.0518	0.4956	0.144*
C06	0.8339 (8)	−0.0618 (7)	0.53047 (17)	0.083 (2)
H06	0.7472	−0.0705	0.5221	0.124*
C07	0.7650 (8)	−0.2033 (6)	0.57539 (16)	0.078 (2)
H07	0.6790	−0.1986	0.5642	0.117*
H08	0.7953	−0.2895	0.5748	0.117*
O01	0.7467 (5)	−0.1645 (4)	0.60869 (11)	0.0773 (14)
C08	0.6649 (7)	−0.0610 (5)	0.61232 (17)	0.0628 (16)
O08	0.6001 (5)	−0.0148 (4)	0.59100 (11)	0.0698 (11)
N1	0.6688 (5)	−0.0248 (4)	0.64339 (12)	0.0604 (13)
H1	0.7095	−0.0694	0.6576	0.091*
C1A	0.6022 (6)	0.0919 (5)	0.65229 (16)	0.0612 (15)
H1A	0.5104	0.0923	0.6434	0.092*
C1B	0.5937 (8)	0.1048 (7)	0.68870 (17)	0.080 (2)
H1B1	0.5426	0.1798	0.6936	0.119*
H1B2	0.6846	0.1168	0.6969	0.119*
C1G	0.5281 (10)	−0.0066 (9)	0.70728 (18)	0.101 (3)
H1G	0.5837	−0.0804	0.7029	0.152*
C1D	0.5401 (12)	0.0205 (13)	0.7438 (2)	0.153 (5)
H1D1	0.4955	0.0978	0.7487	0.229*
H1D2	0.6339	0.0265	0.7496	0.229*
H1D3	0.4985	−0.0461	0.7557	0.229*
C1E	0.3885 (12)	−0.0360 (11)	0.6959 (3)	0.147 (4)
H1E1	0.3857	−0.0324	0.6729	0.220*
H1E2	0.3261	0.0237	0.7047	0.220*
H1E3	0.3638	−0.1184	0.7030	0.220*
C1	0.6803 (7)	0.2001 (6)	0.63834 (16)	0.0620 (16)
O1	0.7937 (5)	0.2259 (4)	0.64992 (13)	0.0800 (14)
N2	0.6290 (5)	0.2601 (4)	0.61332 (11)	0.0576 (13)
H2	0.5459	0.2473	0.6081	0.086*
C2A	0.7101 (6)	0.3486 (6)	0.59397 (14)	0.0582 (15)
C2L	0.8129 (7)	0.2714 (7)	0.57435 (18)	0.080 (2)
H2L1	0.7807	0.2619	0.5527	0.121*
H2L2	0.8235	0.1905	0.5840	0.121*
H2L3	0.8986	0.3136	0.5741	0.121*
C2R	0.6146 (7)	0.4189 (7)	0.57078 (16)	0.0750 (19)
H2R1	0.6140	0.3776	0.5503	0.113*
H2R2	0.6458	0.5032	0.5681	0.113*
H2R3	0.5247	0.4196	0.5795	0.113*
C2	0.7832 (7)	0.4438 (5)	0.61476 (13)	0.0563 (15)
O2	0.8998 (4)	0.4776 (4)	0.60746 (9)	0.0583 (10)
N3	0.7218 (4)	0.4969 (4)	0.63985 (10)	0.0510 (11)
C3A	0.7918 (6)	0.5955 (5)	0.65803 (13)	0.0544 (14)
H3A	0.8118	0.6650	0.6434	0.082*
C3B	0.6866 (7)	0.6393 (7)	0.68293 (17)	0.080 (2)
H3B1	0.6977	0.5953	0.7032	0.120*
H3B2	0.6940	0.7284	0.6868	0.120*
C3G	0.5529 (7)	0.6072 (7)	0.66734 (19)	0.083 (2)
H3G1	0.4821	0.5994	0.6834	0.124*
H3G2	0.5269	0.6708	0.6519	0.124*
C3D	0.5772 (6)	0.4845 (6)	0.65084 (17)	0.0680 (17)
H3D1	0.5165	0.4732	0.6327	0.102*
H3D2	0.5665	0.4150	0.6656	0.102*
C3	0.9212 (7)	0.5564 (6)	0.67459 (14)	0.0590 (16)
O3	0.9980 (5)	0.6391 (4)	0.68455 (11)	0.0747 (13)
N4	0.9439 (6)	0.4351 (4)	0.67884 (12)	0.0642 (14)
H4	0.8860	0.3826	0.6713	0.096*
C4A	1.0614 (7)	0.3877 (6)	0.69554 (15)	0.0715 (18)
H4A	1.1066	0.4571	0.7065	0.107*
C4B	1.0176 (9)	0.2879 (9)	0.72121 (16)	0.098 (3)
H4B	0.9756	0.2174	0.7100	0.147*
C4G	0.9148 (12)	0.3439 (12)	0.7439 (2)	0.148 (5)
H4G1	0.8322	0.3598	0.7325	0.223*
H4G2	0.9493	0.4209	0.7525	0.223*
H4G3	0.8976	0.2866	0.7612	0.223*
C4C	1.1318 (13)	0.2416 (15)	0.7397 (2)	0.171 (6)
H4C1	1.1868	0.1890	0.7263	0.257*
H4C2	1.0992	0.1943	0.7578	0.257*
H4C3	1.1847	0.3108	0.7473	0.257*
C4	1.1592 (8)	0.3290 (6)	0.67218 (16)	0.0723 (18)
O4	1.2807 (6)	0.3377 (7)	0.67326 (14)	0.112 (2)
O51	1.0956 (4)	0.2598 (4)	0.65013 (10)	0.0653 (11)
C51	1.1792 (8)	0.1917 (7)	0.62804 (17)	0.079 (2)
H511	1.1260	0.1250	0.6186	0.118*
H512	1.2529	0.1535	0.6399	0.118*
C5	1.2378 (7)	0.2700 (6)	0.60140 (16)	0.0665 (16)
O5	1.3462 (5)	0.2397 (4)	0.58902 (12)	0.0812 (13)
N5	1.1648 (5)	0.3693 (5)	0.59226 (12)	0.0635 (13)
H5	1.0868	0.3811	0.6009	0.095*
C5A	1.2129 (6)	0.4583 (6)	0.56810 (14)	0.0638 (16)
H5A	1.2966	0.4235	0.5592	0.096*
C61	1.2497 (6)	0.5822 (6)	0.58316 (14)	0.0618 (15)
C62	1.3333 (7)	0.6648 (6)	0.56631 (17)	0.0730 (18)
H62	1.3632	0.6435	0.5458	0.110*
C63	1.3721 (7)	0.7769 (7)	0.5795 (2)	0.081 (2)
H63	1.4286	0.8299	0.5679	0.121*
C64	1.3293 (8)	0.8111 (7)	0.6089 (2)	0.086 (2)
H64	1.3553	0.8879	0.6174	0.128*
C65	1.2455 (8)	0.7311 (8)	0.62706 (18)	0.085 (2)
H65	1.2165	0.7538	0.6475	0.128*
C66	1.2072 (7)	0.6177 (7)	0.61361 (17)	0.0752 (19)
H66	1.1517	0.5643	0.6253	0.113*
C71	1.1145 (6)	0.4673 (6)	0.54079 (15)	0.0608 (16)
C72	1.1090 (8)	0.3707 (7)	0.51915 (18)	0.083 (2)
H72	1.1665	0.3027	0.5217	0.124*
C73	1.0184 (9)	0.3726 (8)	0.49326 (19)	0.096 (2)
H73	1.0124	0.3045	0.4794	0.143*
C74	0.9375 (8)	0.4768 (9)	0.48838 (18)	0.088 (2)
H74	0.8784	0.4799	0.4710	0.132*
C75	0.9453 (8)	0.5730 (8)	0.50889 (19)	0.090 (2)
H75	0.8913	0.6430	0.5056	0.135*
C76	1.0337 (7)	0.5697 (7)	0.53530 (16)	0.0761 (19)
H76	1.0377	0.6375	0.5493	0.114*
O	0.9222 (5)	−0.1052 (3)	0.67743 (17)	0.142 (3)
H1W	0.982 (3)	−0.042 (3)	0.6695 (16)	0.171*
H2W	0.961 (6)	−0.182 (2)	0.670 (2)	0.171*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
C01	0.083 (4)	0.051 (3)	0.053 (3)	−0.001 (3)	−0.004 (3)	−0.003 (3)
C02	0.092 (5)	0.079 (5)	0.072 (4)	0.003 (5)	−0.020 (4)	0.007 (4)
C03	0.073 (5)	0.113 (6)	0.107 (6)	−0.006 (5)	−0.005 (5)	−0.002 (6)
C04	0.086 (5)	0.093 (5)	0.089 (5)	−0.003 (5)	0.003 (4)	0.008 (5)
C05	0.111 (6)	0.108 (6)	0.069 (5)	−0.013 (6)	−0.004 (4)	0.020 (5)
C06	0.085 (5)	0.086 (5)	0.078 (5)	−0.007 (5)	−0.008 (4)	0.006 (4)
C07	0.101 (5)	0.059 (4)	0.073 (4)	−0.002 (4)	0.021 (4)	−0.010 (4)
O01	0.104 (4)	0.061 (2)	0.066 (3)	0.023 (3)	0.012 (3)	0.008 (2)
C08	0.065 (4)	0.042 (3)	0.081 (5)	−0.010 (3)	0.010 (4)	0.003 (3)
O08	0.082 (3)	0.056 (2)	0.072 (3)	0.004 (2)	−0.011 (2)	0.001 (2)
N1	0.076 (3)	0.048 (3)	0.056 (3)	0.005 (3)	0.003 (3)	0.004 (2)
C1A	0.058 (3)	0.052 (3)	0.074 (4)	0.000 (3)	0.003 (3)	−0.003 (3)
C1B	0.088 (5)	0.071 (4)	0.080 (5)	0.004 (4)	0.006 (4)	−0.001 (4)
C1G	0.134 (7)	0.091 (5)	0.078 (5)	0.033 (6)	0.035 (5)	0.010 (5)
C1D	0.191 (11)	0.191 (12)	0.077 (5)	−0.010 (11)	0.034 (6)	0.009 (7)
C1E	0.152 (10)	0.149 (10)	0.140 (9)	−0.051 (9)	0.025 (8)	0.027 (8)
C1	0.065 (4)	0.053 (3)	0.069 (4)	0.002 (3)	−0.001 (3)	0.000 (3)
O1	0.071 (3)	0.057 (2)	0.112 (4)	−0.003 (2)	−0.026 (3)	0.014 (3)
N2	0.055 (3)	0.057 (3)	0.062 (3)	−0.011 (2)	−0.005 (2)	0.005 (3)
C2A	0.058 (3)	0.059 (3)	0.058 (3)	−0.004 (3)	0.000 (3)	−0.002 (3)
C2L	0.070 (4)	0.077 (4)	0.095 (5)	−0.010 (4)	0.017 (4)	−0.029 (4)
C2R	0.078 (4)	0.080 (4)	0.067 (4)	−0.018 (4)	−0.007 (3)	0.012 (4)
C2	0.073 (4)	0.048 (3)	0.048 (3)	0.003 (3)	−0.005 (3)	0.005 (3)
O2	0.050 (2)	0.056 (2)	0.069 (2)	−0.0131 (19)	0.0075 (19)	0.000 (2)
N3	0.048 (3)	0.049 (2)	0.056 (3)	0.001 (2)	0.001 (2)	−0.006 (2)
C3A	0.063 (4)	0.046 (3)	0.054 (3)	0.001 (3)	−0.001 (3)	0.007 (3)
C3B	0.079 (5)	0.072 (4)	0.087 (5)	0.007 (4)	0.003 (4)	−0.025 (4)
C3G	0.065 (4)	0.089 (5)	0.094 (5)	0.018 (4)	0.009 (4)	−0.015 (4)
C3D	0.049 (4)	0.075 (4)	0.080 (4)	0.004 (3)	0.014 (3)	−0.003 (4)
C3	0.069 (4)	0.059 (4)	0.049 (3)	−0.002 (4)	0.000 (3)	−0.003 (3)
O3	0.081 (3)	0.053 (2)	0.090 (3)	−0.011 (2)	−0.018 (3)	−0.009 (3)
N4	0.075 (3)	0.048 (3)	0.069 (3)	0.009 (3)	−0.016 (3)	0.006 (3)
C4A	0.084 (4)	0.068 (4)	0.063 (4)	0.011 (4)	−0.021 (4)	0.000 (3)
C4B	0.116 (6)	0.126 (6)	0.053 (4)	0.056 (6)	0.002 (4)	0.019 (5)
C4G	0.175 (10)	0.170 (10)	0.101 (7)	0.081 (9)	0.033 (7)	0.053 (7)
C4C	0.190 (11)	0.221 (14)	0.103 (7)	0.092 (12)	0.004 (8)	0.039 (9)
C4	0.084 (5)	0.068 (4)	0.065 (4)	0.007 (4)	−0.007 (4)	0.010 (4)
O4	0.066 (3)	0.143 (5)	0.126 (5)	−0.008 (4)	−0.012 (3)	0.002 (4)
O51	0.063 (2)	0.066 (2)	0.067 (3)	0.009 (2)	−0.002 (2)	0.000 (2)
C51	0.092 (5)	0.067 (4)	0.078 (5)	0.016 (4)	0.011 (4)	0.010 (4)
C5	0.074 (4)	0.055 (3)	0.070 (4)	0.012 (4)	0.000 (4)	−0.002 (3)
O5	0.064 (3)	0.081 (3)	0.098 (3)	0.023 (3)	0.010 (3)	0.005 (3)
N5	0.057 (3)	0.062 (3)	0.071 (3)	0.006 (3)	0.011 (3)	0.013 (3)
C5A	0.060 (4)	0.064 (4)	0.067 (4)	0.006 (3)	0.009 (3)	0.012 (3)
C61	0.059 (4)	0.066 (4)	0.061 (4)	−0.001 (3)	−0.005 (3)	0.003 (3)
C62	0.069 (4)	0.076 (4)	0.074 (4)	−0.004 (4)	−0.001 (3)	0.004 (4)
C63	0.067 (4)	0.071 (4)	0.104 (6)	−0.018 (4)	−0.008 (4)	0.015 (5)
C64	0.078 (5)	0.078 (5)	0.100 (6)	−0.004 (5)	−0.021 (5)	−0.010 (5)
C65	0.087 (5)	0.090 (5)	0.078 (5)	−0.007 (5)	−0.010 (4)	−0.007 (4)
C66	0.076 (4)	0.076 (4)	0.074 (4)	−0.015 (4)	−0.008 (4)	0.002 (4)
C71	0.057 (3)	0.059 (4)	0.066 (4)	−0.005 (3)	0.009 (3)	0.000 (3)
C72	0.085 (5)	0.072 (4)	0.090 (5)	0.012 (4)	0.008 (4)	−0.026 (4)
C73	0.114 (6)	0.093 (6)	0.079 (5)	−0.009 (6)	−0.008 (5)	−0.019 (5)
C74	0.090 (5)	0.105 (6)	0.069 (5)	−0.012 (5)	−0.003 (4)	0.008 (5)
C75	0.090 (5)	0.092 (5)	0.087 (5)	0.024 (5)	−0.021 (5)	−0.001 (5)
C76	0.083 (5)	0.075 (4)	0.070 (4)	0.015 (4)	−0.005 (4)	−0.001 (4)
O	0.174 (6)	0.105 (4)	0.148 (6)	0.040 (5)	−0.076 (5)	−0.026 (4)

Geometric parameters (Å, º)

C01—C06	1.366 (9)	C3G—C3D	1.501 (9)
C01—C02	1.383 (10)	C3G—H3G1	0.9700
C01—C07	1.507 (9)	C3G—H3G2	0.9700
C02—C03	1.388 (11)	C3D—H3D1	0.9700
C02—H02	0.9300	C3D—H3D2	0.9700
C03—C04	1.337 (11)	C3—O3	1.237 (7)
C03—H03	0.9300	C3—N4	1.329 (8)
C04—C05	1.394 (11)	N4—C4A	1.445 (8)
C04—H04	0.9300	N4—H4	0.8600
C05—C06	1.379 (10)	C4A—C4	1.507 (10)
C05—H05	0.9300	C4A—C4B	1.571 (10)
C06—H06	0.9300	C4A—H4A	0.9800
C07—O01	1.456 (8)	C4B—C4C	1.453 (13)
C07—H07	0.9700	C4B—C4G	1.510 (11)
C07—H08	0.9700	C4B—H4B	0.9800
O01—C08	1.380 (7)	C4G—H4G1	0.9600
C08—O08	1.200 (7)	C4G—H4G2	0.9600
C08—N1	1.349 (8)	C4G—H4G3	0.9600
N1—C1A	1.459 (7)	C4C—H4C1	0.9600
N1—H1	0.8600	C4C—H4C2	0.9600
C1A—C1	1.508 (9)	C4C—H4C3	0.9600
C1A—C1B	1.522 (9)	C4—O4	1.205 (8)
C1A—H1A	0.9800	C4—O51	1.335 (8)
C1B—C1G	1.561 (11)	O51—C51	1.434 (8)
C1B—H1B1	0.9700	C51—C5	1.505 (9)
C1B—H1B2	0.9700	C51—H511	0.9700
C1G—C1E	1.491 (13)	C51—H512	0.9700
C1G—C1D	1.549 (11)	C5—O5	1.232 (8)
C1G—H1G	0.9800	C5—N5	1.340 (8)
C1D—H1D1	0.9600	N5—C5A	1.464 (7)
C1D—H1D2	0.9600	N5—H5	0.8600
C1D—H1D3	0.9600	C5A—C71	1.497 (9)
C1E—H1E1	0.9600	C5A—C61	1.511 (9)
C1E—H1E2	0.9600	C5A—H5A	0.9800
C1E—H1E3	0.9600	C61—C66	1.387 (9)
C1—O1	1.251 (7)	C61—C62	1.398 (9)
C1—N2	1.323 (7)	C62—C63	1.373 (10)
N2—C2A	1.479 (7)	C62—H62	0.9300
N2—H2	0.8600	C63—C64	1.345 (10)
C2A—C2	1.519 (8)	C63—H63	0.9300
C2A—C2L	1.543 (9)	C64—C65	1.409 (11)
C2A—C2R	1.545 (9)	C64—H64	0.9300
C2L—H2L1	0.9600	C65—C66	1.389 (10)
C2L—H2L2	0.9600	C65—H65	0.9300
C2L—H2L3	0.9600	C66—H66	0.9300
C2R—H2R1	0.9600	C71—C72	1.372 (9)
C2R—H2R2	0.9600	C71—C76	1.375 (9)
C2R—H2R3	0.9600	C72—C73	1.400 (11)
C2—O2	1.246 (7)	C72—H72	0.9300
C2—N3	1.334 (7)	C73—C74	1.387 (11)
N3—C3A	1.471 (7)	C73—H73	0.9300
N3—C3D	1.506 (7)	C74—C75	1.339 (11)
C3A—C3	1.512 (8)	C74—H74	0.9300
C3A—C3B	1.539 (9)	C75—C76	1.403 (9)
C3A—H3A	0.9800	C75—H75	0.9300
C3B—C3G	1.512 (10)	C76—H76	0.9300
C3B—H3B1	0.9700	O—H1W	0.96 (11)
C3B—H3B2	0.9700	O—H2W	0.96 (6)
C06—C01—C02	118.8 (7)	H3B1—C3B—H3B2	109.0
C06—C01—C07	120.7 (7)	C3D—C3G—C3B	104.7 (6)
C02—C01—C07	120.5 (6)	C3D—C3G—H3G1	110.8
C01—C02—C03	118.9 (7)	C3B—C3G—H3G1	110.8
C01—C02—H02	120.5	C3D—C3G—H3G2	110.8
C03—C02—H02	120.5	C3B—C3G—H3G2	110.8
C04—C03—C02	123.0 (8)	H3G1—C3G—H3G2	108.9
C04—C03—H03	118.5	C3G—C3D—N3	102.3 (5)
C02—C03—H03	118.5	C3G—C3D—H3D1	111.3
C03—C04—C05	117.9 (8)	N3—C3D—H3D1	111.3
C03—C04—H04	121.0	C3G—C3D—H3D2	111.3
C05—C04—H04	121.0	N3—C3D—H3D2	111.3
C06—C05—C04	120.2 (8)	H3D1—C3D—H3D2	109.2
C06—C05—H05	119.9	O3—C3—N4	123.5 (6)
C04—C05—H05	119.9	O3—C3—C3A	118.2 (5)
C01—C06—C05	121.1 (7)	N4—C3—C3A	118.3 (6)
C01—C06—H06	119.4	C3—N4—C4A	122.8 (6)
C05—C06—H06	119.4	C3—N4—H4	118.6
O01—C07—C01	111.3 (5)	C4A—N4—H4	118.6
O01—C07—H07	109.4	N4—C4A—C4	110.7 (5)
C01—C07—H07	109.4	N4—C4A—C4B	110.1 (6)
O01—C07—H08	109.4	C4—C4A—C4B	109.3 (6)
C01—C07—H08	109.4	N4—C4A—H4A	108.9
H07—C07—H08	108.0	C4—C4A—H4A	108.9
C08—O01—C07	113.9 (5)	C4B—C4A—H4A	108.9
O08—C08—N1	127.1 (6)	C4C—C4B—C4G	109.1 (7)
O08—C08—O01	124.3 (6)	C4C—C4B—C4A	112.2 (8)
N1—C08—O01	108.6 (6)	C4G—C4B—C4A	109.9 (7)
C08—N1—C1A	118.4 (5)	C4C—C4B—H4B	108.5
C08—N1—H1	120.8	C4G—C4B—H4B	108.5
C1A—N1—H1	120.8	C4A—C4B—H4B	108.5
N1—C1A—C1	109.3 (5)	C4B—C4G—H4G1	109.5
N1—C1A—C1B	110.8 (5)	C4B—C4G—H4G2	109.5
C1—C1A—C1B	109.9 (5)	H4G1—C4G—H4G2	109.5
N1—C1A—H1A	108.9	C4B—C4G—H4G3	109.5
C1—C1A—H1A	108.9	H4G1—C4G—H4G3	109.5
C1B—C1A—H1A	108.9	H4G2—C4G—H4G3	109.5
C1A—C1B—C1G	116.4 (6)	C4B—C4C—H4C1	109.5
C1A—C1B—H1B1	108.2	C4B—C4C—H4C2	109.5
C1G—C1B—H1B1	108.2	H4C1—C4C—H4C2	109.5
C1A—C1B—H1B2	108.2	C4B—C4C—H4C3	109.5
C1G—C1B—H1B2	108.2	H4C1—C4C—H4C3	109.5
H1B1—C1B—H1B2	107.3	H4C2—C4C—H4C3	109.5
C1E—C1G—C1D	114.8 (8)	O4—C4—O51	122.5 (7)
C1E—C1G—C1B	112.9 (8)	O4—C4—C4A	125.7 (7)
C1D—C1G—C1B	108.0 (9)	O51—C4—C4A	111.8 (6)
C1E—C1G—H1G	106.9	C4—O51—C51	116.7 (6)
C1D—C1G—H1G	106.9	O51—C51—C5	114.2 (5)
C1B—C1G—H1G	106.9	O51—C51—H511	108.7
C1G—C1D—H1D1	109.5	C5—C51—H511	108.7
C1G—C1D—H1D2	109.5	O51—C51—H512	108.7
H1D1—C1D—H1D2	109.5	C5—C51—H512	108.7
C1G—C1D—H1D3	109.5	H511—C51—H512	107.6
H1D1—C1D—H1D3	109.5	O5—C5—N5	124.0 (6)
H1D2—C1D—H1D3	109.5	O5—C5—C51	119.7 (6)
C1G—C1E—H1E1	109.5	N5—C5—C51	116.3 (6)
C1G—C1E—H1E2	109.5	C5—N5—C5A	122.4 (5)
H1E1—C1E—H1E2	109.5	C5—N5—H5	118.8
C1G—C1E—H1E3	109.5	C5A—N5—H5	118.8
H1E1—C1E—H1E3	109.5	N5—C5A—C71	110.6 (5)
H1E2—C1E—H1E3	109.5	N5—C5A—C61	111.5 (5)
O1—C1—N2	122.6 (6)	C71—C5A—C61	114.4 (5)
O1—C1—C1A	118.7 (6)	N5—C5A—H5A	106.6
N2—C1—C1A	118.7 (6)	C71—C5A—H5A	106.6
C1—N2—C2A	122.0 (5)	C61—C5A—H5A	106.6
C1—N2—H2	119.0	C66—C61—C62	117.6 (6)
C2A—N2—H2	119.0	C66—C61—C5A	123.1 (6)
N2—C2A—C2	112.2 (5)	C62—C61—C5A	119.4 (6)
N2—C2A—C2L	107.5 (5)	C63—C62—C61	121.2 (7)
C2—C2A—C2L	110.3 (5)	C63—C62—H62	119.4
N2—C2A—C2R	108.7 (5)	C61—C62—H62	119.4
C2—C2A—C2R	108.6 (5)	C64—C63—C62	120.8 (7)
C2L—C2A—C2R	109.5 (6)	C64—C63—H63	119.6
C2A—C2L—H2L1	109.5	C62—C63—H63	119.6
C2A—C2L—H2L2	109.5	C63—C64—C65	120.4 (7)
H2L1—C2L—H2L2	109.5	C63—C64—H64	119.8
C2A—C2L—H2L3	109.5	C65—C64—H64	119.8
H2L1—C2L—H2L3	109.5	C66—C65—C64	118.4 (7)
H2L2—C2L—H2L3	109.5	C66—C65—H65	120.8
C2A—C2R—H2R1	109.5	C64—C65—H65	120.8
C2A—C2R—H2R2	109.5	C61—C66—C65	121.6 (7)
H2R1—C2R—H2R2	109.5	C61—C66—H66	119.2
C2A—C2R—H2R3	109.5	C65—C66—H66	119.2
H2R1—C2R—H2R3	109.5	C72—C71—C76	118.0 (6)
H2R2—C2R—H2R3	109.5	C72—C71—C5A	118.3 (6)
O2—C2—N3	119.2 (5)	C76—C71—C5A	123.7 (6)
O2—C2—C2A	119.8 (5)	C71—C72—C73	121.2 (7)
N3—C2—C2A	120.9 (6)	C71—C72—H72	119.4
C2—N3—C3A	119.6 (5)	C73—C72—H72	119.4
C2—N3—C3D	129.1 (5)	C74—C73—C72	119.5 (7)
C3A—N3—C3D	110.7 (5)	C74—C73—H73	120.2
N3—C3A—C3	115.6 (5)	C72—C73—H73	120.2
N3—C3A—C3B	104.2 (5)	C75—C74—C73	119.5 (7)
C3—C3A—C3B	110.5 (5)	C75—C74—H74	120.2
N3—C3A—H3A	108.8	C73—C74—H74	120.2
C3—C3A—H3A	108.8	C74—C75—C76	120.9 (7)
C3B—C3A—H3A	108.8	C74—C75—H75	119.5
C3G—C3B—C3A	103.5 (5)	C76—C75—H75	119.5
C3G—C3B—H3B1	111.1	C71—C76—C75	120.8 (7)
C3A—C3B—H3B1	111.1	C71—C76—H76	119.6
C3G—C3B—H3B2	111.1	C75—C76—H76	119.6
C3A—C3B—H3B2	111.1	H1W—O—H2W	104 (4)
C06—C01—C02—C03	−0.6 (10)	C4B—C4A—C4—O51	79.2 (7)
C07—C01—C02—C03	−179.7 (6)	O4—C4—O51—C51	2.1 (9)
C01—C02—C03—C04	−2.0 (12)	C4A—C4—O51—C51	−175.5 (5)
C02—C03—C04—C05	3.4 (13)	C4—O51—C51—C5	−78.0 (7)
C03—C04—C05—C06	−2.3 (13)	O51—C51—C5—O5	151.9 (6)
C02—C01—C06—C05	1.6 (11)	O51—C51—C5—N5	−29.2 (9)
C07—C01—C06—C05	−179.3 (7)	O5—C5—N5—C5A	−5.2 (10)
C04—C05—C06—C01	−0.2 (13)	C51—C5—N5—C5A	176.0 (6)
C06—C01—C07—O01	−124.8 (7)	C5—N5—C5A—C71	122.8 (6)
C02—C01—C07—O01	54.3 (8)	C5—N5—C5A—C61	−108.8 (6)
C01—C07—O01—C08	78.6 (7)	N5—C5A—C61—C66	−17.1 (8)
C07—O01—C08—O08	8.9 (8)	C71—C5A—C61—C66	109.3 (7)
C07—O01—C08—N1	−172.6 (5)	N5—C5A—C61—C62	161.2 (6)
O08—C08—N1—C1A	−8.6 (9)	C71—C5A—C61—C62	−72.5 (8)
O01—C08—N1—C1A	173.0 (5)	C66—C61—C62—C63	0.1 (10)
C08—N1—C1A—C1	−69.6 (7)	C5A—C61—C62—C63	−178.3 (6)
C08—N1—C1A—C1B	169.2 (6)	C61—C62—C63—C64	−0.7 (11)
N1—C1A—C1B—C1G	−53.4 (8)	C62—C63—C64—C65	1.1 (11)
C1—C1A—C1B—C1G	−174.3 (6)	C63—C64—C65—C66	−0.8 (11)
C1A—C1B—C1G—C1E	−56.5 (10)	C62—C61—C66—C65	0.1 (10)
C1A—C1B—C1G—C1D	175.4 (8)	C5A—C61—C66—C65	178.5 (6)
N1—C1A—C1—O1	−70.5 (8)	C64—C65—C66—C61	0.2 (10)
C1B—C1A—C1—O1	51.2 (8)	N5—C5A—C71—C72	−74.5 (7)
C1B—C1A—C1—N2	−132.1 (6)	C61—C5A—C71—C72	158.7 (6)
O1—C1—N2—C2A	9.8 (9)	N5—C5A—C71—C76	108.7 (7)
N1—C1A—C1—N2	106.2 (6)	C61—C5A—C71—C76	−18.1 (9)
C1A—C1—N2—C2A	−166.8 (5)	C76—C71—C72—C73	−3.6 (11)
C1—N2—C2A—C2	−50.0 (7)	C5A—C71—C72—C73	179.3 (7)
C1—N2—C2A—C2L	71.5 (7)	C71—C72—C73—C74	3.4 (12)
C1—N2—C2A—C2R	−170.1 (5)	C72—C73—C74—C75	−1.4 (12)
N2—C2A—C2—O2	141.9 (5)	C73—C74—C75—C76	−0.2 (13)
C2L—C2A—C2—O2	22.0 (7)	C72—C71—C76—C75	2.0 (10)
C2R—C2A—C2—O2	−97.9 (6)	C5A—C71—C76—C75	178.8 (7)
N2—C2A—C2—N3	−41.6 (7)	C74—C75—C76—C71	−0.1 (12)
C2L—C2A—C2—N3	−161.5 (5)	O01—C08—N1—C1A	173.0 (5)
C2R—C2A—C2—N3	78.5 (7)	C08—N1—C1A—C1	−69.6 (7)
O2—C2—N3—C3A	1.0 (7)	N1—C1A—C1—N2	106.2 (6)
C2A—C2—N3—C3A	−175.5 (5)	C1A—C1—N2—C2A	−166.8 (5)
O2—C2—N3—C3D	171.5 (5)	C1—N2—C2A—C2	−50.0 (7)
C2A—C2—N3—C3D	−5.0 (9)	N2—C2A—C2—N3	−41.6 (7)
C2—N3—C3A—C3	−62.6 (6)	C2A—C2—N3—C3A	−175.5 (5)
C3D—N3—C3A—C3	125.3 (5)	C2—N3—C3A—C3	−62.6 (6)
C2—N3—C3A—C3B	176.0 (5)	N3—C3A—C3—N4	−17.2 (7)
C3D—N3—C3A—C3B	3.9 (6)	C3A—C3—N4—C4A	−177.4 (5)
N3—C3A—C3B—C3G	−25.4 (6)	C3—N4—C4A—C4	−107.6 (7)
C3—C3A—C3B—C3G	−150.2 (6)	N4—C4A—C4—O51	−42.2 (7)
C3A—C3B—C3G—C3D	38.0 (7)	C4A—C4—O51—C51	−175.5 (5)
C3B—C3G—C3D—N3	−35.0 (7)	C4—O51—C51—C5	−78.0 (7)
C2—N3—C3D—C3G	−151.9 (6)	O51—C51—C5—N5	−29.2 (9)
C3A—N3—C3D—C3G	19.2 (7)	C51—C5—N5—C5A	176.0 (6)
N3—C3A—C3—O3	165.5 (5)	C5—N5—C5A—C71	122.8 (6)
C3B—C3A—C3—O3	−76.4 (7)	C5—N5—C5A—C61	−108.8 (6)
N3—C3A—C3—N4	−17.2 (7)	N1—C1A—C1B—C1G	−53.4 (8)
C3B—C3A—C3—N4	100.9 (7)	C1A—C1B—C1G—C1E	−56.5 (10)
O3—C3—N4—C4A	−0.3 (10)	C1A—C1B—C1G—C1D	175.4 (8)
C3A—C3—N4—C4A	−177.4 (5)	N3—C3A—C3B—C3G	−25.4 (6)
C3—N4—C4A—C4	−107.6 (7)	C3—C3A—C3B—C3G	−150.2 (6)
C3—N4—C4A—C4B	131.4 (7)	C3A—C3B—C3G—C3D	38.0 (7)
N4—C4A—C4B—C4C	−177.3 (8)	C3B—C3G—C3D—N3	−35.0 (7)
C4—C4A—C4B—C4C	60.9 (10)	C3A—N3—C3D—C3G	19.2 (7)
N4—C4A—C4B—C4G	−55.8 (9)	C2—N3—C3D—C3G	−151.9 (6)
C4—C4A—C4B—C4G	−177.5 (8)	C3D—N3—C3A—C3B	3.9 (6)
N4—C4A—C4—O4	140.4 (7)	N4—C4A—C4B—C4C	−177.3 (8)
C4B—C4A—C4—O4	−98.2 (9)	N4—C4A—C4B—C4G	−55.8 (9)
N4—C4A—C4—O51	−42.2 (7)

Hydrogen-bond geometry (Å, º)

D—H···A	D—H	H···A	D···A	D—H···A
N4—H4···O1	0.86	2.11	2.944 (7)	164
N5—H5···O2	0.86	2.13	2.932 (7)	154
N1—H1···O	0.86	2.29	3.002 (7)	140
N2—H2···O5^i	0.86	2.13	2.979 (7)	170
O—H2W···O3^ii	0.96 (6)	2.04 (4)	2.853 (6)	142 (6)

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