N-Z-Pro-D-Leu using synchrotron radiation data from a very small crystal

The crystal structure of the neuroactive artificial dipeptide N-­benzyl­oxy­carbonylprolyl-D-leucine, C₁₉H₂₆N₂O₅, was solved using synchrotron radiation data collected on a very small crystal (20 × 20 × 380 [mu]m). The mol­ecules form hydrogen-bonded 2₁ helices. The acid carbonyl group does not participate in strong hydrogen bonds. This is interpreted as a consequence of close-packing requirements.

S1. Comment

N-benzyloxycarbonyl-prolyl-D-leucine (NZPL) is a synthetic dipeptide protected on the N-terminal side by a benzyloxycarbonyl (Z) group. The compound is effective as inhibitor of the development of tolerance to and physical dependence on morphine in mice (Walter et al., 1978, 1979). It appears to influence the brain stem concentration of norandrenaline and dopamine (Kovács et al., 1981, 1983, 1984) showing that its function is linked to the neurotransmitter system of the brain. Furthermore, Szabó et al. (1987) found that NZPL attenuates the development of tolerance to the hypothermic effect of ethanol. \sch

Here we solve the crystal structure of NZPL using synchrotron radiation data collected at the Swiss-Norwegian Beam Line (SNBL) at the European Synchrotron Radiation Facility, France. SNBL is situated at a bending magnet. A MAR345 imaging plate system and focusing optics were employed for the measurements. The crystal was very small and measured only 20×20×380µm. Despite this small size, data of satisfying quality could be collected and the structure solved and refined, see the refinement statistics.

The molecular backbone is bent at C2A and ϕ₂ is 112.6 (3)° (see Fig. 1). The Pro residue adopts the envelope conformation, C1G is in the N1A–C1A–C1D plane [Δ = -0.001 (11) Å] while C1B is 0.540 (9) Å below it. This is also reflected in the torsion angles (Table 1). There appears to be somewhat reduced delocalization over the peptide bond between Z and Pro as seen by comparing the bond lengths (C═O, C–N) with those of the delocalized peptide link between Pro and D-Leu.

The most interesting feature of the present structure is the imbalance between the number of donors and acceptors of classical hydrogen bonds. There are three hydrogen-bond acceptors (O02, O1 and O2') but only two strong donors (O2''–H2'' and N2–H2). Surprisingly, the structure does not form carboxylic acid dimers. O2' does not accept any classical hydrogen bonds; see Table 2. The carboxylic acid moiety is a donor in a O–H···O hydrogen bond with O1ⁱ. Together with the remaining strong hydrogen bonds (Table 2), this leads to helical 2₁ columns along the a axis as illustrated in Fig. 2. Further stabilization of this columnar structure is provided by the C—H···O contacts (Table 2). One of these is to O2' while the second is to O1. The latter is the shortest as would be expected from the slightly higher acidity of CO2 than of C2B. In addition to these intermolecular contacts, there is a very short intramolecular C—H···O contact: C2A—H2A···O1 making a C₅ ring. It is unclear whether this interaction is attractive or not. The fact that carboxylic acid dimers or catamers do not form and that the acid carbonyl oxygen doess not accept strong hydrogen bonds, is certainly due to close-packing requirements and the hydrogen-bond donor-acceptor imbalance. This demonstrates that close-packing is the principal factor governing crystal structures.

The study of small crystals using synchrotron radiation has been reviewed by Harding (1988, 1995, 1996), Rieck & Schulz (1991), Clegg (2000) and Birkedal (2000). Two measures of scattering powers have been proposed. Rieck et al. (1988) suggest using S = (F(000)/V_cell)²V_crystalλ³ while Harding (1988) used S' = V_crystal(Σ f_i²/V_primitive²), where V_crystal is the sample volume, V_primitive is the volume of the primitive unit cell and Σf_i² is the sum of the squares of the atomic numbers over the primitive unit cell. With these definitions we get S = 1.2^.10 ¹⁶ e² and S' = 1.7^.10 ¹⁴ e² Å^-3. These scattering powers lie between those accessible with laboratory equipment and those requiring dedicated microfocus beamlines. They are of the same order of magnitude as those considered belonging to the class of very small crystals by Harding (1996). The present data set extends to 0.9 Å and as many as 88% of the measured reflections are observed at the I>2σ(I) level. This exceeds the typical qualities quoted by Harding (1996). Note that the present sample is needle-shaped. Clearly, it is the smallest dimension that effectively defines a crystal to be of micro size. A cube-shaped sample with side lengths of 20 µm yielding scattering powers S = 3.2^.10 ¹⁴ e² and S' = 9.1^.10 ¹² e² Å^-3, which is typical of the micro-crystal domain (Birkedal, 2000), could have been measured with the installation of SNBL. In the present experiment, the exposure time was 5 s per image. This resulted in several saturated low order reflections and a second data set was collected with an attenuating filter in the incident beam. To measure a crystal of 20 times smaller volume would thus correspond to an exposure time of 100 s per image. This would have increased the measuring time from 130 min to only 270 min for a complete data set of high quality. This small increase in measuring time reflects the fact that a large part of the experiment time is used for detector read out (Birkedal, 2000). These considerations demonstrate that the present setup would be quite capable of measuring purely organic samples in the 20×20×20 µm range. This result opens up some interesting experimental possibilities on bending magnet beamlines at third generation synchrotrons for microcrystal diffraction, hitherto considered to be an exclusive domain of dedicated insertion device beamlines.

Related literature

For related literature, see: Birkedal (2000); Clegg (2000); Harding (1988, 1995, 1996); Kovács et al. (1981, 1983, 1984); Otwinowski & Minor (1997); Rieck & Schulz (1991); Rieck et al. (1988); Szabó et al. (1987); Walter et al. (1978, 1979).

S2. Experimental

A single-crystal was selected directly from the commercially acquired batch (SIGMA C4644, lot 23 F5900) and mounted on a thin glass needle. The size was estimated from optical microscopy images at 80× magnification. The s.u.'s given in the Crystal Data table reflect our estimated confidence in these numbers.

Two data collections were performed: HIGH and LOW. Dataset HIGH was done with a crystal-to-detector distance of 130 mm and a 5 s exposure time. Dataset LOW was done with a 50 µm Cu-filter in the beam, a crystal-to-detector distance of 180 mm and 20 s exposure time. This second data set was collected to compensate for the limited dynamic range of the detector and effectively corresponds to a measurement of the strong low-order reflections. For both data sets, 90 images were collected. A 2° oscillation range was used for all images, which roughly corresponds to a hemisphere of data. Before the data collection started, we verified on a test image that none of the symmetry elements of the crystal were parallel to the oscillation axis to ensure as complete a data set as possible. The final data set was more than 96% complete. The beam size, selected with the MAR345 receiving slits, was 0.5×0.5 mm. The degree of linear polarization was assumed to be 0.96. This value was found to be valid for the present set-up under conditions similar to the present (Birkedal, 2000). The mosaic spread of the crystal was somewhat non-uniform and orientation dependent. The chosen peak shape is a compromise between the small spot sizes found on some images and the larger ones found on others. The data were corrected for changes in the incident beam intensity by an interframe scaling procedure as implemented in SCALEPACK (Otwinowski & Minor, 1997). By comparing the scale factors of individual frames of data set HIGH with the corresponding ones of LOW, we determined the effective attenuation factor of the incident beam Cu absorber foil to be 17.9 (3).

S3. Refinement

Due to the lack of chiral resolving power in the experiment (no sizeable anomalous scattering contribution), Friedel mates were averaged. The enantiomer was chosen so that the peptide had the known chirality. Thus the Flack (1983) parameter is not a valuable descriptor of absolute configuration.

Computing details

Cell refinement: HKL (Otwinowski & Minor, 1997); data reduction: HKL; program(s) used to solve structure: SHELXS97 (Sheldrick, 1997); program(s) used to refine structure: SHELXL97 (Sheldrick, 1997); molecular graphics: XP (Siemens, 1996).

Figures

	Fig. 1. Molecular structure and labeling scheme of NZPL. Displacement ellipsoids are drawn at the 50% probability level.
	Fig. 2. The helical hydrogen-bonded column in NZPL. Projection along the b axis. Dashed lines indicate N—H···O and O—H···O hydrogen bonds while dotted lines represent C—H···O close contacts.

N-benzyloxycarbonyl-prolyl-D-leucine

Crystal data

C19H26N2O5	Dx = 1.211 Mg m^−3
Mr = 362.42	Synchrotron radiation, λ = 0.8008 Å
Orthorhombic, P212121	Cell parameters from 6811 reflections
a = 6.8870 (14) Å	θ = 2.0–26.4°
b = 12.851 (3) Å	µ = 0.09 mm^−1
c = 22.462 (5) Å	T = 293 K
V = 1988.0 (7) Å^3	Needle, colourless
Z = 4	0.38 (1) × 0.02 (1) × 0.02 (1) mm
F(000) = 776

Data collection

MAR345 diffractometer	1413 reflections with I > 2σ(I)
Radiation source: bending magnet 1 at ESRF	Rint = 0.045
Si(111) double crystal monochromator with bent second crystal for sagital focusing	θmax = 26.4°, θmin = 2.0°
Detector resolution: 6.667 pixels mm^-1	h = −7→7
ϕ–scans	k = −14→14
11028 measured reflections	l = −24→24
1613 independent reflections

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)] = 0.036	H atoms treated by a mixture of independent and constrained refinement
wR(F^2) = 0.104	w = 1/[σ^2(Fo^2) + (0.0549P)^2 + 0.2963P] where P = (Fo^2 + 2Fc^2)/3
S = 1.05	(Δ/σ)max < 0.001
1613 reflections	Δρmax = 0.11 e Å^−3
238 parameters	Δρmin = −0.10 e Å^−3
0 restraints	Absolute structure: Fixed by known peptide configuration
Primary atom site location: structure-invariant direct methods	Absolute structure parameter: 1 (2)

Special details

Experimental. The wavelength was calibrated with a standard Si powder pattern.

All reflections were involved in a global scaling procedure to correct for beam decay (the measurements were performed on a synchrotron) and inhomogeneities (in beam, sample mount absorption etc.). In data set HIGH, scales varied between 7.3 (3) and 12.1 (5). In data set LOW the variation was between 1.66 (7) and 2.7 (2).

Cell parameters were determined in a refinement of cell parameters, setting angles and mosaicity after scaling (Otwinowski & Minor, 1997). This procedures uses strong reflections from the entire data set.

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
O01	0.2632 (3)	0.58333 (19)	0.16786 (10)	0.0766 (7)
O02	0.5397 (3)	0.6565 (2)	0.20278 (10)	0.0812 (8)
C01	0.3619 (5)	0.6525 (3)	0.20036 (14)	0.0670 (9)
C02	0.3729 (5)	0.5193 (3)	0.12706 (16)	0.0801 (10)
H02A	0.4740	0.4820	0.1482	0.096*
H02B	0.4331	0.5621	0.0967	0.096*
C03	0.2337 (5)	0.4443 (3)	0.09933 (15)	0.0711 (9)
C04	0.2773 (6)	0.3398 (3)	0.09514 (17)	0.0873 (11)
H04	0.3964	0.3154	0.1088	0.105*
C05	0.1447 (8)	0.2708 (4)	0.07064 (19)	0.1008 (14)
H05	0.1745	0.2003	0.0687	0.121*
C06	−0.0293 (8)	0.3056 (4)	0.04939 (19)	0.1007 (14)
H06	−0.1174	0.2593	0.0326	0.121*
C07	−0.0730 (7)	0.4099 (4)	0.05309 (17)	0.0926 (12)
H07	−0.1915	0.4342	0.0389	0.111*
C08	0.0572 (6)	0.4782 (3)	0.07765 (17)	0.0803 (10)
H08	0.0261	0.5486	0.0797	0.096*
O1	0.1489 (3)	0.80960 (19)	0.12792 (9)	0.0698 (6)
N1	0.2413 (4)	0.7140 (2)	0.23164 (11)	0.0694 (8)
C1A	0.0351 (4)	0.7249 (3)	0.21672 (13)	0.0692 (9)
H1A	−0.0329	0.6583	0.2210	0.083*
C1	0.0116 (4)	0.7675 (3)	0.15429 (12)	0.0575 (8)
C1B	−0.0329 (6)	0.8027 (4)	0.26379 (16)	0.1045 (16)
H1B1	−0.1438	0.8422	0.2497	0.125*
H1B2	−0.0678	0.7675	0.3005	0.125*
C1G	0.1396 (7)	0.8717 (5)	0.2727 (2)	0.1186 (17)
H1G1	0.1370	0.9029	0.3120	0.142*
H1G2	0.1419	0.9267	0.2432	0.142*
C1D	0.3132 (6)	0.8020 (3)	0.26603 (16)	0.0879 (12)
H1D1	0.4171	0.8374	0.2450	0.106*
H1D2	0.3607	0.7795	0.3046	0.106*
O2'	−0.2950 (4)	0.6449 (2)	0.03463 (10)	0.0778 (7)
O2"	−0.2699 (4)	0.78285 (19)	−0.02518 (9)	0.0740 (7)
H2"	−0.2976	0.7404	−0.0511	0.111*
N2	−0.1662 (3)	0.7619 (2)	0.13110 (10)	0.0588 (7)
H2	−0.2526	0.7260	0.1496	0.071*
C2	−0.2645 (4)	0.7357 (3)	0.02740 (14)	0.0585 (8)
C2A	−0.2210 (4)	0.8132 (2)	0.07640 (13)	0.0569 (8)
H2A	−0.1102	0.8554	0.0634	0.068*
C2B	−0.3931 (4)	0.8869 (3)	0.08607 (15)	0.0635 (9)
H2B1	−0.4284	0.9169	0.0480	0.076*
H2B2	−0.5029	0.8462	0.0998	0.076*
C2G	−0.3591 (6)	0.9748 (3)	0.13000 (18)	0.0828 (11)
H2G	−0.3295	0.9436	0.1688	0.099*
C2D1	−0.1887 (8)	1.0423 (4)	0.1125 (3)	0.159 (3)
H2D1	−0.2157	1.0757	0.0751	0.239*
H2D2	−0.0746	0.9999	0.1086	0.239*
H2D3	−0.1674	1.0942	0.1425	0.239*
C2D2	−0.5421 (7)	1.0394 (3)	0.1370 (2)	0.1132 (16)
H2D4	−0.6483	0.9949	0.1479	0.170*
H2D5	−0.5712	1.0736	0.1001	0.170*
H2D6	−0.5226	1.0906	0.1676	0.170*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
O01	0.0491 (12)	0.1036 (17)	0.0773 (14)	−0.0041 (14)	0.0032 (12)	−0.0113 (14)
O02	0.0440 (13)	0.122 (2)	0.0778 (15)	−0.0062 (14)	−0.0049 (11)	0.0185 (15)
C01	0.0500 (18)	0.093 (2)	0.0576 (18)	−0.0074 (19)	−0.0062 (16)	0.0166 (19)
C02	0.063 (2)	0.094 (2)	0.083 (2)	0.007 (2)	0.014 (2)	0.007 (2)
C03	0.072 (2)	0.081 (2)	0.0607 (19)	0.005 (2)	0.0070 (18)	0.0103 (18)
C04	0.093 (3)	0.093 (3)	0.076 (2)	0.019 (2)	0.009 (2)	0.004 (2)
C05	0.139 (4)	0.081 (3)	0.082 (3)	0.008 (3)	0.004 (3)	−0.003 (2)
C06	0.129 (4)	0.091 (3)	0.082 (3)	−0.013 (3)	−0.016 (3)	0.001 (2)
C07	0.102 (3)	0.095 (3)	0.081 (2)	−0.006 (3)	−0.016 (2)	0.011 (2)
C08	0.085 (3)	0.074 (2)	0.082 (2)	−0.001 (2)	−0.007 (2)	0.012 (2)
O1	0.0444 (11)	0.1150 (18)	0.0499 (11)	−0.0140 (12)	0.0010 (10)	0.0012 (12)
N1	0.0485 (14)	0.110 (2)	0.0498 (14)	0.0003 (16)	−0.0083 (12)	−0.0029 (15)
C1A	0.0469 (17)	0.109 (3)	0.0519 (17)	−0.0012 (18)	−0.0001 (14)	0.0012 (19)
C1	0.0418 (16)	0.085 (2)	0.0459 (15)	−0.0001 (16)	−0.0024 (13)	−0.0052 (16)
C1B	0.079 (3)	0.182 (5)	0.053 (2)	0.027 (3)	0.0046 (19)	−0.013 (3)
C1G	0.108 (3)	0.157 (4)	0.091 (3)	0.015 (4)	−0.005 (3)	−0.052 (3)
C1D	0.082 (3)	0.119 (3)	0.062 (2)	−0.007 (2)	−0.0177 (19)	−0.007 (2)
O2'	0.0883 (17)	0.0785 (16)	0.0667 (14)	−0.0019 (14)	−0.0023 (13)	−0.0001 (13)
O2"	0.0757 (15)	0.0921 (16)	0.0541 (13)	0.0018 (14)	−0.0080 (12)	0.0043 (13)
N2	0.0391 (13)	0.0858 (18)	0.0516 (14)	−0.0047 (13)	−0.0019 (10)	0.0065 (13)
C2	0.0361 (15)	0.082 (2)	0.0572 (19)	0.0055 (16)	−0.0024 (13)	0.0039 (19)
C2A	0.0414 (15)	0.073 (2)	0.0566 (17)	−0.0046 (15)	−0.0037 (13)	0.0083 (16)
C2B	0.0548 (18)	0.068 (2)	0.0676 (19)	0.0032 (15)	−0.0063 (15)	−0.0025 (17)
C2G	0.084 (2)	0.075 (2)	0.090 (3)	−0.002 (2)	−0.012 (2)	−0.011 (2)
C2D1	0.116 (4)	0.098 (3)	0.264 (8)	−0.031 (3)	0.007 (5)	−0.045 (4)
C2D2	0.106 (3)	0.101 (3)	0.132 (4)	0.019 (3)	−0.004 (3)	−0.029 (3)

Geometric parameters (Å, º)

O01—C01	1.336 (4)	C1B—H1B2	0.9700
O01—C02	1.445 (4)	C1G—C1D	1.501 (6)
O02—C01	1.227 (4)	C1G—H1G1	0.9700
C01—N1	1.345 (4)	C1G—H1G2	0.9700
C02—C03	1.495 (5)	C1D—H1D1	0.9700
C02—H02A	0.9700	C1D—H1D2	0.9700
C02—H02B	0.9700	O2'—C2	1.197 (4)
C03—C08	1.380 (5)	O2"—C2	1.328 (3)
C03—C04	1.380 (5)	O2"—H2"	0.8200
C04—C05	1.387 (6)	N2—C2A	1.444 (4)
C04—H04	0.9300	N2—H2	0.8600
C05—C06	1.365 (7)	C2—C2A	1.514 (5)
C05—H05	0.9300	C2A—C2B	1.533 (4)
C06—C07	1.377 (6)	C2A—H2A	0.9800
C06—H06	0.9300	C2B—C2G	1.519 (5)
C07—C08	1.371 (6)	C2B—H2B1	0.9700
C07—H07	0.9300	C2B—H2B2	0.9700
C08—H08	0.9300	C2G—C2D1	1.512 (6)
O1—C1	1.240 (3)	C2G—C2D2	1.517 (6)
N1—C1D	1.457 (5)	C2G—H2G	0.9800
N1—C1A	1.466 (4)	C2D1—H2D1	0.9600
C1A—C1	1.514 (4)	C2D1—H2D2	0.9600
C1A—C1B	1.529 (5)	C2D1—H2D3	0.9600
C1A—H1A	0.9800	C2D2—H2D4	0.9600
C1—N2	1.333 (3)	C2D2—H2D5	0.9600
C1B—C1G	1.496 (7)	C2D2—H2D6	0.9600
C1B—H1B1	0.9700
C01—O01—C02	117.3 (3)	C1D—C1G—H1G1	110.7
O02—C01—O01	124.1 (4)	C1B—C1G—H1G2	110.7
O02—C01—N1	124.7 (4)	C1D—C1G—H1G2	110.7
O01—C01—N1	111.2 (3)	H1G1—C1G—H1G2	108.8
O01—C02—C03	107.2 (3)	N1—C1D—C1G	104.2 (3)
O01—C02—H02A	110.3	N1—C1D—H1D1	110.9
C03—C02—H02A	110.3	C1G—C1D—H1D1	110.9
O01—C02—H02B	110.3	N1—C1D—H1D2	110.9
C03—C02—H02B	110.3	C1G—C1D—H1D2	110.9
H02A—C02—H02B	108.5	H1D1—C1D—H1D2	108.9
C08—C03—C04	118.4 (4)	C2—O2"—H2"	109.5
C08—C03—C02	120.6 (4)	C1—N2—C2A	123.2 (2)
C04—C03—C02	121.1 (4)	C1—N2—H2	118.4
C03—C04—C05	120.4 (4)	C2A—N2—H2	118.4
C03—C04—H04	119.8	O2'—C2—O2"	124.1 (3)
C05—C04—H04	119.8	O2'—C2—C2A	125.2 (3)
C06—C05—C04	120.5 (4)	O2"—C2—C2A	110.6 (3)
C06—C05—H05	119.7	N2—C2A—C2	111.8 (3)
C04—C05—H05	119.7	N2—C2A—C2B	111.3 (2)
C05—C06—C07	119.3 (5)	C2—C2A—C2B	110.9 (2)
C05—C06—H06	120.3	N2—C2A—H2A	107.6
C07—C06—H06	120.3	C2—C2A—H2A	107.6
C08—C07—C06	120.3 (5)	C2B—C2A—H2A	107.6
C08—C07—H07	119.8	C2G—C2B—C2A	115.7 (3)
C06—C07—H07	119.8	C2G—C2B—H2B1	108.4
C07—C08—C03	121.1 (4)	C2A—C2B—H2B1	108.4
C07—C08—H08	119.5	C2G—C2B—H2B2	108.4
C03—C08—H08	119.5	C2A—C2B—H2B2	108.4
C01—N1—C1D	121.6 (3)	H2B1—C2B—H2B2	107.4
C01—N1—C1A	122.4 (3)	C2D1—C2G—C2D2	111.0 (3)
C1D—N1—C1A	112.1 (3)	C2D1—C2G—C2B	112.2 (4)
N1—C1A—C1	110.5 (2)	C2D2—C2G—C2B	110.3 (3)
N1—C1A—C1B	101.6 (3)	C2D1—C2G—H2G	107.7
C1—C1A—C1B	111.8 (3)	C2D2—C2G—H2G	107.7
N1—C1A—H1A	110.9	C2B—C2G—H2G	107.7
C1—C1A—H1A	110.9	C2G—C2D1—H2D1	109.5
C1B—C1A—H1A	110.9	C2G—C2D1—H2D2	109.5
O1—C1—N2	122.5 (3)	H2D1—C2D1—H2D2	109.5
O1—C1—C1A	121.2 (2)	C2G—C2D1—H2D3	109.5
N2—C1—C1A	116.2 (3)	H2D1—C2D1—H2D3	109.5
C1G—C1B—C1A	103.7 (3)	H2D2—C2D1—H2D3	109.5
C1G—C1B—H1B1	111.0	C2G—C2D2—H2D4	109.5
C1A—C1B—H1B1	111.0	C2G—C2D2—H2D5	109.5
C1G—C1B—H1B2	111.0	H2D4—C2D2—H2D5	109.5
C1A—C1B—H1B2	111.0	C2G—C2D2—H2D6	109.5
H1B1—C1B—H1B2	109.0	H2D4—C2D2—H2D6	109.5
C1B—C1G—C1D	105.4 (4)	H2D5—C2D2—H2D6	109.5
C1B—C1G—H1G1	110.7
C02—O01—C01—O02	−8.8 (5)	C1B—C1A—C1—O1	96.2 (4)
C02—O01—C01—N1	173.6 (3)	N1—C1A—C1—N2	167.3 (3)
C01—O01—C02—C03	176.3 (3)	C1B—C1A—C1—N2	−80.4 (4)
O01—C02—C03—C08	46.4 (4)	N1—C1A—C1B—C1G	33.7 (4)
O01—C02—C03—C04	−133.1 (3)	C1—C1A—C1B—C1G	−84.1 (4)
C08—C03—C04—C05	−1.1 (6)	C1A—C1B—C1G—C1D	−35.1 (5)
C02—C03—C04—C05	178.4 (3)	C01—N1—C1D—C1G	158.1 (4)
C03—C04—C05—C06	1.1 (6)	C1A—N1—C1D—C1G	0.0 (4)
C04—C05—C06—C07	−0.7 (7)	C1B—C1G—C1D—N1	22.0 (4)
C05—C06—C07—C08	0.3 (7)	O1—C1—N2—C2A	−6.6 (5)
C06—C07—C08—C03	−0.3 (6)	C1A—C1—N2—C2A	169.9 (3)
C04—C03—C08—C07	0.7 (6)	C1—N2—C2A—C2	112.6 (3)
C02—C03—C08—C07	−178.8 (4)	C1—N2—C2A—C2B	−122.9 (3)
O02—C01—N1—C1D	8.4 (5)	O2'—C2—C2A—N2	16.3 (4)
O01—C01—N1—C1D	−174.0 (3)	O2"—C2—C2A—N2	−165.5 (2)
O02—C01—N1—C1A	164.3 (3)	O2'—C2—C2A—C2B	−108.5 (4)
O01—C01—N1—C1A	−18.2 (4)	O2"—C2—C2A—C2B	69.7 (3)
C01—N1—C1A—C1	−60.3 (5)	N2—C2A—C2B—C2G	60.7 (4)
C1D—N1—C1A—C1	97.6 (3)	C2—C2A—C2B—C2G	−174.2 (3)
C01—N1—C1A—C1B	−179.0 (3)	C2A—C2B—C2G—C2D1	57.7 (5)
C1D—N1—C1A—C1B	−21.2 (4)	C2A—C2B—C2G—C2D2	−178.0 (3)
N1—C1A—C1—O1	−16.1 (5)

Hydrogen-bond geometry (Å, º)

D—H···A	D—H	H···A	D···A	D—H···A
O2"—H2"···O1^i	0.82	1.88	2.655 (3)	158
N2—H2···O02^ii	0.86	2.07	2.920 (3)	172
C2B—H2B2···O1^ii	0.97	2.52	3.438 (4)	157
C02—H02B···O2′^iii	0.97	2.57	3.486 (4)	158
C2A—H2A···O1	0.98	2.37	2.798 (3)	106

Symmetry codes: (i) x−1/2, −y+3/2, −z; (ii) x−1, y, z; (iii) x+1, y, z.