How does binding of imidazole-based inhibitors to heme oxygenase-1 influence their conformation? Insights combining crystal structures and molecular modelling

Heme oxygenase-1 (HO-1) inhibition is associated with antitumor activity. Imidazole-based analogues show effective and selective inhibitory potency of HO-1. In this work, five single-crystal structures of four imidazole-based compounds are presented, with an in-depth structural analysis. In order to study the influence of the conformation of the ligands on binding to protein, conformational data from crystallography are compared with quantum mechanics analysis and molecular docking studies. Molecular docking of imidazole-based analogues in the active site of HO-1 is in good agreement with the experimental structures. Inhibitors interact with the heme cofactor and a hydrophobic pocket (Met34, Phe37, Val50, Leu147 and Phe214) in the HO-1 binding site. An alternate binding mode can be hypothesized for some inhibitors in the series.

Computing details

Data collection: CrysAlis PRO, Agilent Technologies, Version 1.171.36.28 (release 01-02-2013 CrysAlis171 .NET) (compiled Feb 1 2013,16:14:44) for 1aHClH2O, 1bHClH2O, 1c3H2O; CrysAlis PRO, Agilent Technologies, Version 1.171.35.21 (release 20-01-2012 CrysAlis171 .NET) (compiled Jan 23 2012,18:06:46) for 2aHCl. Cell refinement: CrysAlis PRO, Agilent Technologies, Version 1.171.36.28 (release 01-02-2013 CrysAlis171 .NET) (compiled Feb 1 2013,16:14:44) for 1aHClH2O, 1bHClH2O, 1c3H2O; CrysAlis PRO, Agilent Technologies, Version 1.171.35.21 (release 20-01-2012 CrysAlis171 .NET) (compiled Jan 23 2012,18:06:46) for (1b), 2aHCl. Data reduction: CrysAlis PRO, Agilent Technologies, Version 1.171.36.28 (release 01-02-2013 CrysAlis171 .NET) (compiled Feb 1 2013,16:14:44) for 1aHClH2O, 1bHClH2O, 1c3H2O; CrysAlis PRO, Agilent Technologies, Version 1.171.35.21 (release 20-01-2012 CrysAlis171 .NET) (compiled Jan 23 2012,18:06:46) for (1b), 2aHCl. For all compounds, program(s) used to refine structure: SHELXL97 (Sheldrick, 1997).

Figures

(1aHClH2O)

Crystal data

C11H11N2O·Cl·H2O	Dx = 1.347 Mg m^−3
Mr = 240.68	Mo Kα radiation, λ = 0.71073 Å
Orthorhombic, Pcab	Cell parameters from 895 reflections
a = 5.1830 (6) Å	θ = 4.2–21.3°
b = 16.808 (7) Å	µ = 0.31 mm^−1
c = 27.247 (3) Å	T = 293 K
V = 2373.7 (11) Å^3	Needle-like, colourless
Z = 8	0.35 × 0.15 × 0.05 mm
F(000) = 1008

Data collection

Xcalibur, Ruby, Gemini ultra diffractometer	2083 independent reflections
Radiation source: fine-focus sealed tube	1332 reflections with I > 2σ(I)
Graphite monochromator	Rint = 0.076
Detector resolution: 10.3712 pixels mm^-1	θmax = 25.0°, θmin = 3.9°
ω scans	h = −5→6
Absorption correction: multi-scan CrysAlis PRO, Agilent Technologies, Version 1.171.36.28 (release 01-02-2013 CrysAlis171 .NET) (compiled Feb 1 2013,16:14:44) Empirical absorption correction using spherical harmonics, implemented in SCALE3 ABSPACK scaling algorithm.	k = −19→12
Tmin = 0.946, Tmax = 0.985	l = −28→32
6343 measured reflections

Refinement

Refinement on F^2	Primary atom site location: structure-invariant direct methods
Least-squares matrix: full	Secondary atom site location: difference Fourier map
R[F^2 > 2σ(F^2)] = 0.061	Hydrogen site location: inferred from neighbouring sites
wR(F^2) = 0.110	H atoms treated by a mixture of independent and constrained refinement
S = 1.08	w = 1/[σ^2(Fo^2) + (0.0287P)^2] where P = (Fo^2 + 2Fc^2)/3
1986 reflections	(Δ/σ)max < 0.001
153 parameters	Δρmax = 0.19 e Å^−3
3 restraints	Δρmin = −0.16 e Å^−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.

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
O1	0.8942 (4)	0.18538 (15)	0.11090 (7)	0.0541 (7)
N1	0.5206 (4)	0.11706 (18)	0.05702 (9)	0.0463 (7)
C5	0.7475 (5)	0.1448 (2)	0.13474 (10)	0.0374 (8)
C11	0.6126 (5)	0.0862 (2)	0.21558 (11)	0.0462 (9)
H11	0.4779	0.0596	0.2001	0.055*
C7	0.9792 (5)	0.1712 (2)	0.21171 (11)	0.0455 (9)
H7	1.0937	0.2021	0.1936	0.055*
C10	0.6457 (6)	0.0788 (2)	0.26558 (12)	0.0553 (10)
H10	0.5335	0.0473	0.2838	0.066*
C6	0.7787 (5)	0.13299 (19)	0.18821 (10)	0.0342 (7)
C4	0.5233 (5)	0.1041 (2)	0.10980 (10)	0.0483 (9)
H4B	0.5323	0.0474	0.1162	0.058*
H4A	0.3634	0.1238	0.1237	0.058*
C9	0.8433 (6)	0.1178 (3)	0.28833 (12)	0.0552 (10)
H9	0.8642	0.1130	0.3221	0.066*
C8	1.0118 (6)	0.1642 (2)	0.26184 (12)	0.0531 (9)
H8	1.1463	0.1905	0.2775	0.064*
C2	0.3604 (6)	0.1671 (2)	0.03172 (12)	0.0560 (11)
H2	0.2292	0.1981	0.0450	0.067*
N2	0.6292 (5)	0.1116 (2)	−0.01854 (10)	0.0575 (8)
H2N	0.7089	0.0989	−0.0451	0.069*
C3	0.4269 (6)	0.1632 (3)	−0.01550 (13)	0.0591 (10)
H3	0.3501	0.1904	−0.0414	0.071*
C1	0.6837 (6)	0.0841 (2)	0.02569 (12)	0.0563 (11)
H1	0.8135	0.0480	0.0334	0.068*
Cl1	0.96604 (15)	0.03832 (6)	−0.09885 (3)	0.0548 (3)
O2	0.1390 (6)	0.2143 (2)	0.87505 (12)	0.0786 (9)
H2O	−0.004 (4)	0.2391 (19)	0.873 (2)	0.15 (3)*
H2P	0.101 (7)	0.1656 (6)	0.8792 (19)	0.13 (2)*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
O1	0.0528 (12)	0.063 (2)	0.0463 (13)	−0.0127 (12)	0.0053 (11)	0.0107 (12)
N1	0.0432 (14)	0.056 (2)	0.0394 (15)	−0.0027 (14)	−0.0009 (13)	−0.0011 (15)
C5	0.0354 (16)	0.037 (2)	0.0399 (18)	0.0049 (15)	0.0026 (15)	−0.0040 (17)
C11	0.0490 (18)	0.044 (3)	0.046 (2)	−0.0082 (17)	0.0010 (15)	0.0044 (17)
C7	0.0446 (17)	0.044 (2)	0.047 (2)	−0.0029 (16)	0.0051 (15)	0.0012 (17)
C10	0.058 (2)	0.063 (3)	0.044 (2)	−0.0015 (19)	0.0072 (18)	0.0137 (19)
C6	0.0339 (15)	0.026 (2)	0.0423 (17)	0.0025 (14)	0.0033 (14)	−0.0006 (15)
C4	0.0491 (18)	0.059 (3)	0.0364 (18)	−0.0079 (18)	0.0033 (14)	0.0018 (17)
C9	0.066 (2)	0.062 (3)	0.0368 (18)	0.014 (2)	0.0001 (18)	0.0009 (19)
C8	0.0538 (19)	0.055 (3)	0.051 (2)	−0.0039 (19)	−0.0049 (18)	−0.0069 (19)
C2	0.0536 (19)	0.069 (3)	0.045 (2)	0.014 (2)	−0.0051 (17)	−0.004 (2)
N2	0.0719 (18)	0.065 (3)	0.0360 (16)	0.0002 (18)	0.0061 (14)	−0.0077 (16)
C3	0.068 (2)	0.061 (3)	0.049 (2)	0.012 (2)	−0.0121 (18)	0.000 (2)
C1	0.059 (2)	0.068 (3)	0.042 (2)	0.0138 (19)	−0.0019 (18)	−0.005 (2)
Cl1	0.0702 (5)	0.0481 (6)	0.0459 (5)	0.0082 (5)	0.0014 (4)	0.0022 (5)
O2	0.0796 (18)	0.059 (3)	0.097 (2)	−0.0085 (18)	−0.0066 (16)	0.0090 (19)

Geometric parameters (Å, º)

O1—C5	1.211 (3)	C4—H4B	0.9700
N1—C1	1.323 (4)	C4—H4A	0.9700
N1—C2	1.368 (4)	C9—C8	1.375 (5)
N1—C4	1.455 (4)	C9—H9	0.9300
C5—C6	1.479 (4)	C8—H8	0.9300
C5—C4	1.510 (4)	C2—C3	1.334 (4)
C11—C10	1.379 (4)	C2—H2	0.9300
C11—C6	1.384 (4)	N2—C1	1.321 (4)
C11—H11	0.9300	N2—C3	1.363 (4)
C7—C6	1.380 (4)	N2—H2N	0.8600
C7—C8	1.381 (4)	C3—H3	0.9300
C7—H7	0.9300	C1—H1	0.9300
C10—C9	1.365 (4)	O2—H2O	0.8500 (11)
C10—H10	0.9300	O2—H2P	0.8501 (13)
C1—N1—C2	108.7 (3)	C5—C4—H4A	109.1
C1—N1—C4	124.6 (3)	H4B—C4—H4A	107.8
C2—N1—C4	126.6 (3)	C10—C9—C8	120.7 (3)
O1—C5—C6	122.4 (3)	C10—C9—H9	119.7
O1—C5—C4	119.8 (3)	C8—C9—H9	119.7
C6—C5—C4	117.8 (3)	C9—C8—C7	119.4 (3)
C10—C11—C6	120.4 (3)	C9—C8—H8	120.3
C10—C11—H11	119.8	C7—C8—H8	120.3
C6—C11—H11	119.8	C3—C2—N1	107.4 (3)
C6—C7—C8	120.7 (3)	C3—C2—H2	126.3
C6—C7—H7	119.6	N1—C2—H2	126.3
C8—C7—H7	119.6	C1—N2—C3	109.4 (3)
C9—C10—C11	119.9 (3)	C1—N2—H2N	125.3
C9—C10—H10	120.0	C3—N2—H2N	125.3
C11—C10—H10	120.0	C2—C3—N2	106.8 (3)
C7—C6—C11	118.9 (3)	C2—C3—H3	126.6
C7—C6—C5	118.5 (3)	N2—C3—H3	126.6
C11—C6—C5	122.6 (3)	N2—C1—N1	107.8 (3)
N1—C4—C5	112.6 (2)	N2—C1—H1	126.1
N1—C4—H4B	109.1	N1—C1—H1	126.1
C5—C4—H4B	109.1	H2O—O2—H2P	106.24 (19)
N1—C4—H4A	109.1
C6—C11—C10—C9	0.0 (5)	C6—C5—C4—N1	−177.2 (3)
C8—C7—C6—C11	−1.1 (5)	C11—C10—C9—C8	−0.4 (6)
C8—C7—C6—C5	177.9 (3)	C10—C9—C8—C7	0.2 (6)
C10—C11—C6—C7	0.8 (5)	C6—C7—C8—C9	0.6 (5)
C10—C11—C6—C5	−178.2 (3)	C1—N1—C2—C3	0.7 (4)
O1—C5—C6—C7	1.5 (5)	C4—N1—C2—C3	178.0 (3)
C4—C5—C6—C7	−178.6 (3)	N1—C2—C3—N2	−0.7 (4)
O1—C5—C6—C11	−179.5 (3)	C1—N2—C3—C2	0.5 (4)
C4—C5—C6—C11	0.4 (4)	C3—N2—C1—N1	−0.1 (4)
C1—N1—C4—C5	72.4 (4)	C2—N1—C1—N2	−0.4 (4)
C2—N1—C4—C5	−104.5 (4)	C4—N1—C1—N2	−177.8 (3)
O1—C5—C4—N1	2.7 (4)

(1b)

Crystal data

C11H9BrN2O	Dx = 1.696 Mg m^−3
Mr = 265.10	Mo Kα radiation, λ = 0.71073 Å
Orthorhombic, Pbca	Cell parameters from 2524 reflections
a = 10.1389 (7) Å	θ = 3.4–28.1°
b = 7.5691 (5) Å	µ = 3.93 mm^−1
c = 27.0498 (15) Å	T = 293 K
V = 2075.9 (2) Å^3	Needle-like, colourless
Z = 8	0.40 × 0.08 × 0.05 mm
F(000) = 1056

Data collection

Xcalibur, Ruby, Gemini ultra diffractometer	1826 independent reflections
Radiation source: fine-focus sealed tube	1505 reflections with I > 2σ(I)
Graphite monochromator	Rint = 0.034
Detector resolution: 10.3712 pixels mm^-1	θmax = 25.0°, θmin = 3.4°
ω scans	h = −9→12
Absorption correction: multi-scan CrysAlis PRO, Agilent Technologies, Version 1.171.35.21 (release 20-01-2012 CrysAlis171 .NET) (compiled Jan 23 2012,18:06:46) Empirical absorption correction using spherical harmonics, implemented in SCALE3 ABSPACK scaling algorithm.	k = −7→9
Tmin = 0.692, Tmax = 0.822	l = −29→32
7194 measured 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)] = 0.032	H-atom parameters constrained
wR(F^2) = 0.076	w = 1/[σ^2(Fo^2) + (0.0367P)^2 + 0.6352P] where P = (Fo^2 + 2Fc^2)/3
S = 1.07	(Δ/σ)max = 0.001
1821 reflections	Δρmax = 0.43 e Å^−3
137 parameters	Δρmin = −0.27 e Å^−3
0 restraints	Extinction correction: SHELXL, Fc^*=kFc[1+0.001xFc^2λ^3/sin(2θ)]^-1/4
Primary atom site location: structure-invariant direct methods	Extinction coefficient: 0.0030 (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.

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	0.9710 (3)	0.3077 (4)	0.53886 (10)	0.0416 (7)
H1	1.0442	0.3774	0.5319	0.050*
C2	0.8099 (3)	0.1211 (3)	0.52966 (10)	0.0394 (7)
H2	0.7503	0.0401	0.5166	0.047*
C3	0.8128 (3)	0.1835 (4)	0.57615 (10)	0.0445 (7)
H3	0.7544	0.1506	0.6010	0.053*
C4	0.9478 (3)	0.1782 (4)	0.45398 (9)	0.0357 (6)
H4A	0.9439	0.0535	0.4459	0.043*
H4B	1.0381	0.2172	0.4492	0.043*
C5	0.8589 (3)	0.2791 (3)	0.41889 (9)	0.0324 (6)
C6	0.8756 (2)	0.2468 (3)	0.36502 (9)	0.0305 (6)
C7	0.7789 (3)	0.3063 (4)	0.33305 (10)	0.0395 (7)
H7	0.7066	0.3674	0.3455	0.047*
C8	0.7887 (3)	0.2760 (4)	0.28289 (10)	0.0411 (7)
H8	0.7230	0.3159	0.2616	0.049*
C9	0.8958 (3)	0.1869 (3)	0.26456 (9)	0.0357 (6)
C10	0.9941 (3)	0.1265 (3)	0.29573 (10)	0.0386 (7)
H10	1.0666	0.0667	0.2830	0.046*
C11	0.9835 (3)	0.1559 (3)	0.34578 (10)	0.0355 (6)
H11	1.0490	0.1148	0.3669	0.043*
N1	0.9126 (2)	0.2011 (3)	0.50543 (8)	0.0339 (5)
N2	0.9137 (2)	0.3022 (4)	0.58199 (9)	0.0495 (7)
O1	0.7774 (2)	0.3806 (3)	0.43462 (7)	0.0522 (6)
Br1	0.90954 (3)	0.14638 (4)	0.195554 (10)	0.04969 (16)

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
C1	0.0430 (17)	0.0470 (16)	0.0347 (16)	−0.0041 (13)	−0.0094 (14)	0.0014 (13)
C2	0.0307 (15)	0.0510 (16)	0.0366 (15)	0.0001 (12)	0.0001 (12)	0.0029 (13)
C3	0.0418 (18)	0.0593 (18)	0.0323 (15)	0.0071 (15)	0.0028 (13)	0.0046 (14)
C4	0.0362 (15)	0.0452 (15)	0.0257 (13)	0.0047 (12)	0.0001 (12)	−0.0009 (11)
C5	0.0313 (14)	0.0363 (14)	0.0297 (14)	−0.0001 (12)	−0.0006 (12)	−0.0031 (12)
C6	0.0347 (15)	0.0301 (13)	0.0267 (13)	0.0027 (11)	−0.0026 (11)	−0.0004 (11)
C7	0.0437 (18)	0.0421 (14)	0.0326 (14)	0.0133 (13)	−0.0038 (13)	−0.0061 (12)
C8	0.0469 (18)	0.0441 (15)	0.0322 (14)	0.0095 (13)	−0.0090 (13)	−0.0028 (13)
C9	0.0481 (17)	0.0367 (14)	0.0222 (13)	−0.0033 (13)	0.0019 (12)	−0.0021 (11)
C10	0.0385 (16)	0.0424 (15)	0.0348 (15)	0.0043 (12)	0.0062 (13)	−0.0010 (12)
C11	0.0330 (15)	0.0415 (15)	0.0320 (14)	0.0049 (12)	−0.0033 (12)	0.0031 (11)
N1	0.0335 (13)	0.0434 (12)	0.0248 (11)	0.0013 (10)	−0.0021 (9)	−0.0013 (10)
N2	0.0588 (17)	0.0623 (16)	0.0274 (13)	0.0016 (13)	−0.0053 (12)	−0.0052 (12)
O1	0.0575 (14)	0.0650 (13)	0.0342 (11)	0.0278 (11)	−0.0027 (10)	−0.0088 (9)
Br1	0.0671 (3)	0.0558 (2)	0.0262 (2)	0.00046 (15)	0.00192 (13)	−0.00478 (12)

Geometric parameters (Å, º)

C1—N2	1.304 (4)	C5—C6	1.487 (3)
C1—N1	1.349 (3)	C6—C7	1.382 (3)
C1—H1	0.9300	C6—C11	1.394 (4)
C2—C3	1.344 (4)	C7—C8	1.379 (4)
C2—N1	1.372 (3)	C7—H7	0.9300
C2—H2	0.9300	C8—C9	1.372 (4)
C3—N2	1.371 (4)	C8—H8	0.9300
C3—H3	0.9300	C9—C10	1.382 (4)
C4—N1	1.447 (3)	C9—Br1	1.897 (3)
C4—C5	1.516 (4)	C10—C11	1.376 (4)
C4—H4A	0.9700	C10—H10	0.9300
C4—H4B	0.9700	C11—H11	0.9300
C5—O1	1.205 (3)
N2—C1—N1	112.7 (3)	C11—C6—C5	122.4 (2)
N2—C1—H1	123.7	C8—C7—C6	120.7 (2)
N1—C1—H1	123.7	C8—C7—H7	119.7
C3—C2—N1	106.0 (3)	C6—C7—H7	119.7
C3—C2—H2	127.0	C9—C8—C7	119.6 (2)
N1—C2—H2	127.0	C9—C8—H8	120.2
C2—C3—N2	110.8 (3)	C7—C8—H8	120.2
C2—C3—H3	124.6	C8—C9—C10	120.8 (2)
N2—C3—H3	124.6	C8—C9—Br1	119.5 (2)
N1—C4—C5	113.3 (2)	C10—C9—Br1	119.6 (2)
N1—C4—H4A	108.9	C11—C10—C9	119.4 (3)
C5—C4—H4A	108.9	C11—C10—H10	120.3
N1—C4—H4B	108.9	C9—C10—H10	120.3
C5—C4—H4B	108.9	C10—C11—C6	120.5 (2)
H4A—C4—H4B	107.7	C10—C11—H11	119.7
O1—C5—C6	121.9 (2)	C6—C11—H11	119.7
O1—C5—C4	120.5 (2)	C1—N1—C2	106.1 (2)
C6—C5—C4	117.6 (2)	C1—N1—C4	127.5 (2)
C7—C6—C11	119.0 (2)	C2—N1—C4	126.4 (2)
C7—C6—C5	118.6 (2)	C1—N2—C3	104.5 (2)
N1—C2—C3—N2	0.6 (3)	Br1—C9—C10—C11	−179.86 (19)
N1—C4—C5—O1	7.1 (4)	C9—C10—C11—C6	−0.5 (4)
N1—C4—C5—C6	−172.4 (2)	C7—C6—C11—C10	0.3 (4)
O1—C5—C6—C7	−13.2 (4)	C5—C6—C11—C10	179.0 (2)
C4—C5—C6—C7	166.3 (2)	N2—C1—N1—C2	−0.2 (3)
O1—C5—C6—C11	168.0 (3)	N2—C1—N1—C4	178.2 (2)
C4—C5—C6—C11	−12.4 (4)	C3—C2—N1—C1	−0.2 (3)
C11—C6—C7—C8	0.2 (4)	C3—C2—N1—C4	−178.7 (2)
C5—C6—C7—C8	−178.6 (2)	C5—C4—N1—C1	−101.0 (3)
C6—C7—C8—C9	−0.4 (4)	C5—C4—N1—C2	77.1 (3)
C7—C8—C9—C10	0.2 (4)	N1—C1—N2—C3	0.6 (3)
C7—C8—C9—Br1	−179.7 (2)	C2—C3—N2—C1	−0.7 (3)
C8—C9—C10—C11	0.3 (4)

(1bHClH2O)

Crystal data

C11H10BrN2O·Cl·H2O	Z = 2
Mr = 319.58	F(000) = 320
Triclinic, P1	Dx = 1.636 Mg m^−3
a = 6.845 (5) Å	Cu Kα radiation, λ = 1.54178 Å
b = 7.141 (5) Å	Cell parameters from 1215 reflections
c = 13.905 (5) Å	θ = 3.2–65.8°
α = 101.723 (5)°	µ = 6.16 mm^−1
β = 95.164 (5)°	T = 293 K
γ = 100.511 (5)°	Needle-like, colourless
V = 648.6 (7) Å^3	0.40 × 0.25 × 0.10 mm

Data collection

Xcalibur, Ruby, Gemini ultra diffractometer	2355 independent reflections
Radiation source: fine-focus sealed tube	1749 reflections with I > 2σ(I)
Graphite monochromator	Rint = 0.039
Detector resolution: 10.3712 pixels mm^-1	θmax = 67.7°, θmin = 3.3°
ω scans	h = −8→4
Absorption correction: multi-scan CrysAlis PRO, Agilent Technologies, Version 1.171.36.28 (release 01-02-2013 CrysAlis171 .NET) (compiled Feb 1 2013,16:14:44) Empirical absorption correction using spherical harmonics, implemented in SCALE3 ABSPACK scaling algorithm.	k = −8→8
Tmin = 0.197, Tmax = 0.543	l = −16→16
2920 measured reflections

Refinement

Refinement on F^2	Primary atom site location: structure-invariant direct methods
Least-squares matrix: full	Secondary atom site location: difference Fourier map
R[F^2 > 2σ(F^2)] = 0.044	Hydrogen site location: inferred from neighbouring sites
wR(F^2) = 0.121	H atoms treated by a mixture of independent and constrained refinement
S = 1.06	w = 1/[σ^2(Fo^2) + (0.0626P)^2] where P = (Fo^2 + 2Fc^2)/3
2143 reflections	(Δ/σ)max = 0.001
166 parameters	Δρmax = 0.50 e Å^−3
0 restraints	Δρmin = −0.52 e Å^−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.

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
C9	0.2121 (7)	0.2946 (7)	0.4204 (3)	0.0346 (10)
C8	0.2076 (7)	0.0957 (7)	0.4009 (3)	0.0329 (10)
H8	0.1799	0.0214	0.3362	0.039*
C7	0.2446 (6)	0.0103 (7)	0.4782 (3)	0.0321 (10)
H7	0.2435	−0.1229	0.4654	0.039*
C6	0.2840 (6)	0.1182 (6)	0.5762 (3)	0.0305 (10)
C11	0.2854 (7)	0.3166 (7)	0.5947 (3)	0.0357 (11)
H11	0.3104	0.3902	0.6595	0.043*
C10	0.2496 (7)	0.4060 (7)	0.5170 (3)	0.0399 (11)
H10	0.2508	0.5392	0.5294	0.048*
C5	0.3216 (6)	0.0156 (7)	0.6566 (3)	0.0312 (10)
C4	0.3774 (7)	0.1366 (7)	0.7623 (3)	0.0339 (10)
H4A	0.5076	0.2222	0.7688	0.041*
H4B	0.2796	0.2169	0.7780	0.041*
C2	0.2223 (7)	−0.0738 (7)	0.8726 (3)	0.0375 (11)
H2	0.0959	−0.0430	0.8692	0.045*
C3	0.2821 (7)	−0.2050 (7)	0.9191 (4)	0.0431 (12)
H3	0.2059	−0.2809	0.9548	0.052*
C1	0.5354 (7)	−0.0781 (7)	0.8505 (3)	0.0400 (12)
H1	0.6612	−0.0519	0.8300	0.048*
N1	0.3829 (5)	0.0068 (5)	0.8309 (3)	0.0322 (8)
N2	0.4772 (6)	−0.2056 (6)	0.9040 (3)	0.0392 (10)
O1	0.3131 (5)	−0.1593 (5)	0.6410 (2)	0.0425 (8)
O2	0.2485 (7)	0.3911 (8)	1.0329 (4)	0.0662 (14)
Br1	0.16581 (8)	0.42138 (8)	0.31623 (3)	0.0433 (2)
Cl1	−0.06891 (17)	0.28466 (17)	0.84308 (8)	0.0382 (3)
H2N	0.556 (8)	−0.292 (7)	0.928 (4)	0.048 (15)*
H2O	0.201 (9)	0.444 (10)	1.069 (5)	0.06 (2)*
H2P	0.166 (9)	0.351 (9)	0.980 (5)	0.062 (19)*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
C9	0.038 (3)	0.038 (3)	0.030 (2)	0.011 (2)	−0.0003 (19)	0.012 (2)
C8	0.036 (2)	0.033 (3)	0.029 (2)	0.010 (2)	0.0012 (18)	0.0036 (19)
C7	0.032 (2)	0.031 (3)	0.033 (2)	0.0103 (19)	0.0028 (19)	0.0033 (19)
C6	0.024 (2)	0.032 (3)	0.036 (2)	0.0070 (18)	0.0009 (18)	0.0092 (19)
C11	0.050 (3)	0.030 (3)	0.025 (2)	0.010 (2)	−0.002 (2)	0.0046 (19)
C10	0.057 (3)	0.029 (3)	0.032 (2)	0.012 (2)	−0.003 (2)	0.0030 (19)
C5	0.031 (2)	0.031 (3)	0.034 (2)	0.0106 (19)	0.0034 (19)	0.0080 (19)
C4	0.040 (3)	0.031 (2)	0.033 (2)	0.010 (2)	0.000 (2)	0.0130 (19)
C2	0.037 (3)	0.040 (3)	0.042 (3)	0.012 (2)	0.004 (2)	0.019 (2)
C3	0.041 (3)	0.045 (3)	0.047 (3)	0.011 (2)	0.005 (2)	0.018 (2)
C1	0.039 (3)	0.048 (3)	0.040 (3)	0.018 (2)	0.004 (2)	0.017 (2)
N1	0.036 (2)	0.032 (2)	0.0314 (19)	0.0129 (17)	0.0001 (16)	0.0101 (16)
N2	0.049 (2)	0.036 (2)	0.037 (2)	0.0194 (19)	−0.0021 (19)	0.0107 (18)
O1	0.058 (2)	0.0320 (19)	0.0372 (18)	0.0118 (16)	0.0008 (16)	0.0080 (14)
O2	0.072 (3)	0.084 (4)	0.046 (2)	0.055 (3)	−0.011 (2)	−0.005 (2)
Br1	0.0547 (4)	0.0453 (4)	0.0335 (3)	0.0137 (2)	−0.0003 (2)	0.0165 (2)
Cl1	0.0453 (7)	0.0380 (7)	0.0351 (6)	0.0183 (5)	0.0016 (5)	0.0099 (5)

Geometric parameters (Å, º)

C9—C8	1.385 (6)	C4—N1	1.461 (5)
C9—C10	1.390 (7)	C4—H4A	0.9700
C9—Br1	1.890 (4)	C4—H4B	0.9700
C8—C7	1.367 (6)	C2—C3	1.343 (7)
C8—H8	0.9300	C2—N1	1.377 (6)
C7—C6	1.397 (6)	C2—H2	0.9300
C7—H7	0.9300	C3—N2	1.371 (6)
C6—C11	1.386 (6)	C3—H3	0.9300
C6—C5	1.485 (6)	C1—N2	1.318 (6)
C11—C10	1.388 (6)	C1—N1	1.334 (6)
C11—H11	0.9300	C1—H1	0.9300
C10—H10	0.9300	N2—H2N	0.98 (5)
C5—O1	1.213 (5)	O2—H2O	0.70 (6)
C5—C4	1.521 (6)	O2—H2P	0.85 (6)
C8—C9—C10	120.9 (4)	N1—C4—H4A	109.8
C8—C9—Br1	120.8 (3)	C5—C4—H4A	109.8
C10—C9—Br1	118.3 (4)	N1—C4—H4B	109.8
C7—C8—C9	118.9 (4)	C5—C4—H4B	109.8
C7—C8—H8	120.5	H4A—C4—H4B	108.2
C9—C8—H8	120.5	C3—C2—N1	107.2 (4)
C8—C7—C6	121.6 (4)	C3—C2—H2	126.4
C8—C7—H7	119.2	N1—C2—H2	126.4
C6—C7—H7	119.2	C2—C3—N2	106.8 (4)
C11—C6—C7	118.8 (4)	C2—C3—H3	126.6
C11—C6—C5	122.4 (4)	N2—C3—H3	126.6
C7—C6—C5	118.7 (4)	N2—C1—N1	108.2 (4)
C10—C11—C6	120.3 (4)	N2—C1—H1	125.9
C10—C11—H11	119.8	N1—C1—H1	125.9
C6—C11—H11	119.8	C1—N1—C2	108.3 (4)
C11—C10—C9	119.4 (4)	C1—N1—C4	125.1 (4)
C11—C10—H10	120.3	C2—N1—C4	125.8 (4)
C9—C10—H10	120.3	C1—N2—C3	109.4 (4)
O1—C5—C6	122.6 (4)	C1—N2—H2N	127 (3)
O1—C5—C4	119.1 (4)	C3—N2—H2N	124 (3)
C6—C5—C4	118.3 (4)	H2O—O2—H2P	107 (7)
N1—C4—C5	109.6 (4)
C10—C9—C8—C7	−1.2 (7)	C7—C6—C5—C4	176.2 (4)
Br1—C9—C8—C7	179.1 (3)	O1—C5—C4—N1	−8.4 (6)
C9—C8—C7—C6	0.8 (7)	C6—C5—C4—N1	172.9 (3)
C8—C7—C6—C11	0.0 (7)	N1—C2—C3—N2	−1.0 (6)
C8—C7—C6—C5	179.6 (4)	N2—C1—N1—C2	−1.0 (6)
C7—C6—C11—C10	−0.5 (7)	N2—C1—N1—C4	−171.1 (4)
C5—C6—C11—C10	179.9 (4)	C3—C2—N1—C1	1.2 (6)
C6—C11—C10—C9	0.1 (7)	C3—C2—N1—C4	171.3 (4)
C8—C9—C10—C11	0.8 (8)	C5—C4—N1—C1	80.7 (6)
Br1—C9—C10—C11	−179.6 (4)	C5—C4—N1—C2	−87.7 (5)
C11—C6—C5—O1	177.1 (4)	N1—C1—N2—C3	0.4 (6)
C7—C6—C5—O1	−2.5 (6)	C2—C3—N2—C1	0.4 (6)
C11—C6—C5—C4	−4.2 (6)

(1c3H2O)

Crystal data

C11H9N3O3·3(H2O)	Z = 2
Mr = 285.26	F(000) = 300
Triclinic, P1	Dx = 1.405 Mg m^−3
a = 7.0408 (3) Å	Mo Kα radiation, λ = 0.71073 Å
b = 8.4596 (5) Å	Cell parameters from 3247 reflections
c = 11.4455 (7) Å	θ = 3.5–27.0°
α = 89.438 (5)°	µ = 0.12 mm^−1
β = 86.904 (4)°	T = 293 K
γ = 82.072 (4)°	Needle-like, colourless
V = 674.22 (6) Å^3	0.45 × 0.32 × 0.10 mm

Data collection

Xcalibur, Ruby, Gemini ultra diffractometer	2860 independent reflections
Radiation source: fine-focus sealed tube	2238 reflections with I > 2σ(I)
Graphite monochromator	Rint = 0.025
Detector resolution: 10.3712 pixels mm^-1	θmax = 28.1°, θmin = 3.3°
ω scans	h = −9→9
Absorption correction: multi-scan CrysAlis PRO, Agilent Technologies, Version 1.171.36.28 (release 01-02-2013 CrysAlis171 .NET) (compiled Feb 1 2013,16:14:44) Empirical absorption correction using spherical harmonics, implemented in SCALE3 ABSPACK scaling algorithm.	k = −11→10
Tmin = 0.809, Tmax = 1.000	l = −15→14
9754 measured reflections

Refinement

Refinement on F^2	Primary atom site location: structure-invariant direct methods
Least-squares matrix: full	Secondary atom site location: difference Fourier map
R[F^2 > 2σ(F^2)] = 0.046	Hydrogen site location: inferred from neighbouring sites
wR(F^2) = 0.123	H-atom parameters constrained
S = 1.05	w = 1/[σ^2(Fo^2) + (0.0525P)^2 + 0.1757P] where P = (Fo^2 + 2Fc^2)/3
2860 reflections	(Δ/σ)max < 0.001
181 parameters	Δρmax = 0.15 e Å^−3
0 restraints	Δρmin = −0.28 e Å^−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.

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
N1	0.46324 (19)	0.57267 (17)	0.65054 (12)	0.0414 (4)
N2	0.2276 (2)	0.4374 (2)	0.61396 (15)	0.0563 (4)
N3	1.2329 (2)	0.81633 (18)	1.07608 (14)	0.0454 (4)
O1	0.47025 (16)	0.60934 (15)	0.88362 (11)	0.0485 (3)
O2	1.2617 (2)	0.7520 (2)	1.17057 (13)	0.0706 (5)
O3	1.33078 (19)	0.91217 (17)	1.03256 (13)	0.0591 (4)
C1	0.3947 (3)	0.4327 (2)	0.65951 (17)	0.0526 (5)
H1	0.4588	0.3423	0.6943	0.063*
C3	0.1868 (3)	0.5896 (2)	0.57362 (17)	0.0537 (5)
H3	0.0761	0.6294	0.5362	0.064*
C2	0.3289 (3)	0.6744 (2)	0.59545 (17)	0.0515 (5)
H2	0.3346	0.7810	0.5768	0.062*
C4	0.6381 (2)	0.6111 (2)	0.69843 (15)	0.0424 (4)
H4A	0.6730	0.7074	0.6613	0.051*
H4B	0.7417	0.5251	0.6811	0.051*
C5	0.6144 (2)	0.63593 (19)	0.82961 (14)	0.0358 (4)
C6	0.7773 (2)	0.68798 (18)	0.89114 (14)	0.0338 (4)
C7	0.7817 (2)	0.6637 (2)	1.01146 (15)	0.0406 (4)
H7	0.6829	0.6193	1.0511	0.049*
C8	0.9308 (2)	0.7049 (2)	1.07242 (15)	0.0424 (4)
H8	0.9362	0.6863	1.1525	0.051*
C9	1.0720 (2)	0.77451 (19)	1.01137 (15)	0.0371 (4)
C10	1.0712 (2)	0.8032 (2)	0.89326 (15)	0.0414 (4)
H10	1.1675	0.8522	0.8550	0.050*
C11	0.9229 (2)	0.7571 (2)	0.83234 (15)	0.0406 (4)
H11	0.9210	0.7726	0.7518	0.049*
O99	0.1349 (2)	0.12450 (17)	0.62347 (13)	0.0642 (4)
O98	0.4506 (2)	−0.00097 (18)	0.75108 (12)	0.0637 (4)
O97	0.2217 (2)	0.0003 (2)	0.39699 (13)	0.0722 (5)
H99A	0.1543	0.2299	0.6204	0.050*
H97A	0.4667	0.0315	0.8314	0.050*
H98B	0.1248	−0.0565	0.3864	0.050*
H97B	0.5662	0.0090	0.7015	0.050*
H98A	0.1948	0.0377	0.4793	0.050*
H99B	0.2425	0.0856	0.6676	0.050*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
N1	0.0397 (7)	0.0469 (8)	0.0407 (8)	−0.0126 (6)	−0.0110 (6)	−0.0026 (6)
N2	0.0515 (9)	0.0647 (11)	0.0585 (10)	−0.0221 (8)	−0.0183 (8)	−0.0040 (8)
N3	0.0367 (8)	0.0479 (9)	0.0531 (9)	−0.0068 (7)	−0.0099 (7)	−0.0106 (7)
O1	0.0378 (6)	0.0623 (8)	0.0492 (7)	−0.0205 (6)	−0.0024 (5)	0.0014 (6)
O2	0.0667 (9)	0.0919 (12)	0.0602 (9)	−0.0248 (8)	−0.0329 (7)	0.0109 (8)
O3	0.0504 (8)	0.0624 (9)	0.0709 (9)	−0.0267 (7)	−0.0123 (7)	−0.0072 (7)
C1	0.0548 (11)	0.0473 (11)	0.0608 (12)	−0.0178 (9)	−0.0208 (9)	0.0036 (9)
C3	0.0460 (10)	0.0633 (13)	0.0526 (11)	−0.0033 (9)	−0.0194 (9)	−0.0086 (9)
C2	0.0560 (11)	0.0462 (11)	0.0535 (11)	−0.0056 (9)	−0.0189 (9)	−0.0017 (9)
C4	0.0374 (8)	0.0508 (10)	0.0422 (10)	−0.0155 (8)	−0.0067 (7)	−0.0037 (8)
C5	0.0328 (8)	0.0340 (8)	0.0421 (9)	−0.0091 (7)	−0.0047 (7)	0.0017 (7)
C6	0.0301 (8)	0.0339 (8)	0.0381 (9)	−0.0057 (6)	−0.0041 (6)	−0.0022 (7)
C7	0.0377 (9)	0.0461 (10)	0.0402 (9)	−0.0143 (7)	0.0001 (7)	0.0005 (7)
C8	0.0437 (9)	0.0494 (10)	0.0355 (9)	−0.0101 (8)	−0.0057 (7)	−0.0021 (7)
C9	0.0302 (8)	0.0364 (9)	0.0454 (10)	−0.0048 (7)	−0.0079 (7)	−0.0067 (7)
C10	0.0340 (8)	0.0475 (10)	0.0455 (10)	−0.0146 (7)	−0.0029 (7)	0.0031 (8)
C11	0.0369 (8)	0.0509 (10)	0.0362 (9)	−0.0126 (7)	−0.0047 (7)	0.0026 (7)
O99	0.0636 (9)	0.0654 (9)	0.0702 (10)	−0.0253 (7)	−0.0200 (7)	−0.0047 (7)
O98	0.0592 (8)	0.0769 (10)	0.0603 (9)	−0.0240 (7)	−0.0123 (7)	−0.0083 (7)
O97	0.0731 (10)	0.0947 (12)	0.0566 (9)	−0.0379 (9)	−0.0064 (7)	0.0001 (8)

Geometric parameters (Å, º)

N1—C1	1.340 (2)	C6—C11	1.389 (2)
N1—C2	1.365 (2)	C6—C7	1.392 (2)
N1—C4	1.452 (2)	C7—C8	1.377 (2)
N2—C1	1.308 (2)	C7—H7	0.9300
N2—C3	1.362 (3)	C8—C9	1.378 (2)
N3—O2	1.220 (2)	C8—H8	0.9300
N3—O3	1.2206 (19)	C9—C10	1.371 (2)
N3—C9	1.471 (2)	C10—C11	1.387 (2)
O1—C5	1.2066 (19)	C10—H10	0.9300
C1—H1	0.9300	C11—H11	0.9300
C3—C2	1.344 (3)	O99—H99A	0.9205
C3—H3	0.9300	O99—H99B	0.9522
C2—H2	0.9300	O98—H97A	0.9789
C4—C5	1.515 (2)	O98—H97B	0.9798
C4—H4A	0.9700	O97—H98B	0.9014
C4—H4B	0.9700	O97—H98A	0.9967
C5—C6	1.496 (2)
C1—N1—C2	106.24 (15)	O1—C5—C4	120.80 (14)
C1—N1—C4	126.56 (15)	C6—C5—C4	118.35 (13)
C2—N1—C4	127.03 (15)	C11—C6—C7	119.68 (14)
C1—N2—C3	104.60 (15)	C11—C6—C5	122.38 (14)
O2—N3—O3	123.52 (14)	C7—C6—C5	117.94 (14)
O2—N3—C9	117.83 (15)	C8—C7—C6	120.62 (15)
O3—N3—C9	118.64 (15)	C8—C7—H7	119.7
N2—C1—N1	112.50 (17)	C6—C7—H7	119.7
N2—C1—H1	123.8	C7—C8—C9	118.09 (15)
N1—C1—H1	123.8	C7—C8—H8	121.0
C2—C3—N2	110.52 (16)	C9—C8—H8	121.0
C2—C3—H3	124.7	C10—C9—C8	123.15 (14)
N2—C3—H3	124.7	C10—C9—N3	118.71 (14)
C3—C2—N1	106.15 (17)	C8—C9—N3	118.12 (15)
C3—C2—H2	126.9	C9—C10—C11	118.18 (15)
N1—C2—H2	126.9	C9—C10—H10	120.9
N1—C4—C5	111.73 (14)	C11—C10—H10	120.9
N1—C4—H4A	109.3	C10—C11—C6	120.25 (15)
C5—C4—H4A	109.3	C10—C11—H11	119.9
N1—C4—H4B	109.3	C6—C11—H11	119.9
C5—C4—H4B	109.3	H99A—O99—H99B	97.5
H4A—C4—H4B	107.9	H97A—O98—H97B	110.6
O1—C5—C6	120.81 (15)	H98B—O97—H98A	102.3
C3—N2—C1—N1	−0.1 (2)	C11—C6—C7—C8	−1.2 (3)
C2—N1—C1—N2	0.3 (2)	C5—C6—C7—C8	178.12 (15)
C4—N1—C1—N2	175.80 (16)	C6—C7—C8—C9	1.7 (3)
C1—N2—C3—C2	−0.1 (2)	C7—C8—C9—C10	−0.5 (3)
N2—C3—C2—N1	0.3 (2)	C7—C8—C9—N3	−179.09 (15)
C1—N1—C2—C3	−0.3 (2)	O2—N3—C9—C10	−159.83 (17)
C4—N1—C2—C3	−175.82 (17)	O3—N3—C9—C10	19.7 (2)
C1—N1—C4—C5	−75.1 (2)	O2—N3—C9—C8	18.8 (2)
C2—N1—C4—C5	99.5 (2)	O3—N3—C9—C8	−161.64 (16)
N1—C4—C5—O1	6.7 (2)	C8—C9—C10—C11	−1.2 (3)
N1—C4—C5—C6	−175.66 (13)	N3—C9—C10—C11	177.34 (15)
O1—C5—C6—C11	−163.62 (16)	C9—C10—C11—C6	1.8 (3)
C4—C5—C6—C11	18.8 (2)	C7—C6—C11—C10	−0.6 (3)
O1—C5—C6—C7	17.1 (2)	C5—C6—C11—C10	−179.88 (15)
C4—C5—C6—C7	−160.55 (15)

(2aHCl)

Crystal data

C11H12N3O3·Cl	F(000) = 1120
Mr = 269.69	Dx = 1.431 Mg m^−3
Monoclinic, P21/c	Mo Kα radiation, λ = 0.71073 Å
a = 21.9100 (7) Å	Cell parameters from 10843 reflections
b = 8.0131 (3) Å	θ = 3.3–29.3°
c = 14.3802 (4) Å	µ = 0.31 mm^−1
β = 97.325 (3)°	T = 293 K
V = 2504.09 (14) Å^3	Prim, colourless
Z = 8	0.30 × 0.20 × 0.10 mm

Data collection

Xcalibur, Ruby, Gemini ultra diffractometer	4428 independent reflections
Radiation source: fine-focus sealed tube	3979 reflections with I > 2σ(I)
Graphite monochromator	Rint = 0.022
Detector resolution: 10.3712 pixels mm^-1	θmax = 25.0°, θmin = 3.4°
ω scans	h = −26→25
Absorption correction: multi-scan CrysAlis PRO, Agilent Technologies, Version 1.171.35.21 (release 20-01-2012 CrysAlis171 .NET) (compiled Jan 23 2012,18:06:46) Empirical absorption correction using spherical harmonics, implemented in SCALE3 ABSPACK scaling algorithm.	k = −9→9
Tmin = 0.929, Tmax = 0.970	l = −16→17
20723 measured reflections

Refinement

Refinement on F^2	Primary atom site location: structure-invariant direct methods
Least-squares matrix: full	Secondary atom site location: difference Fourier map
R[F^2 > 2σ(F^2)] = 0.038	Hydrogen site location: inferred from neighbouring sites
wR(F^2) = 0.093	H atoms treated by a mixture of independent and constrained refinement
S = 1.05	w = 1/[σ^2(Fo^2) + (0.0364P)^2 + 1.190P] where P = (Fo^2 + 2Fc^2)/3
4418 reflections	(Δ/σ)max < 0.001
341 parameters	Δρmax = 0.23 e Å^−3
0 restraints	Δρmin = −0.24 e Å^−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.

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	0.39014 (10)	−0.0380 (3)	0.45781 (14)	0.0554 (5)
H1	0.3572	0.0327	0.4390	0.066*
C2	0.45540 (10)	−0.2009 (2)	0.54226 (15)	0.0537 (5)
H2	0.4749	−0.2620	0.5925	0.064*
C3	0.47310 (10)	−0.1880 (3)	0.45663 (15)	0.0573 (5)
H3	0.5071	−0.2389	0.4361	0.069*
C4	0.36802 (10)	−0.0821 (3)	0.62132 (14)	0.0547 (5)
H4A	0.3748	−0.1763	0.6638	0.066*
H4B	0.3244	−0.0772	0.5986	0.066*
C5	0.38716 (8)	0.0777 (2)	0.67400 (12)	0.0417 (4)
H5	0.4318	0.0769	0.6921	0.050*
C6	0.35520 (8)	0.0891 (2)	0.76181 (12)	0.0403 (4)
C7	0.37933 (11)	0.0035 (3)	0.84130 (15)	0.0638 (6)
H7	0.4157	−0.0565	0.8411	0.077*
C8	0.35039 (11)	0.0053 (3)	0.92114 (15)	0.0651 (6)
H8	0.3666	−0.0535	0.9744	0.078*
C9	0.29737 (9)	0.0956 (2)	0.92004 (13)	0.0474 (5)
C10	0.27354 (9)	0.1870 (3)	0.84388 (14)	0.0511 (5)
H10	0.2383	0.2510	0.8456	0.061*
C11	0.30273 (9)	0.1828 (3)	0.76414 (13)	0.0477 (4)
H11	0.2868	0.2438	0.7116	0.057*
C21	0.05533 (9)	0.5727 (2)	0.66467 (14)	0.0492 (5)
H21	0.0447	0.5691	0.7252	0.059*
C22	0.09709 (10)	0.5152 (3)	0.53917 (14)	0.0548 (5)
H22	0.1207	0.4639	0.4980	0.066*
C23	0.05931 (10)	0.6451 (3)	0.52014 (15)	0.0615 (6)
H23	0.0518	0.7017	0.4634	0.074*
C24	0.12577 (8)	0.3294 (2)	0.67931 (12)	0.0423 (4)
H24A	0.1273	0.3445	0.7465	0.051*
H24B	0.1677	0.3236	0.6646	0.051*
C25	0.09263 (8)	0.1675 (2)	0.65045 (12)	0.0434 (4)
H25	0.0894	0.1553	0.5822	0.052*
C26	0.12888 (8)	0.0227 (2)	0.69673 (12)	0.0411 (4)
C27	0.16951 (9)	−0.0648 (2)	0.64883 (13)	0.0480 (5)
H27	0.1722	−0.0392	0.5864	0.058*
C28	0.20606 (9)	−0.1895 (3)	0.69241 (14)	0.0523 (5)
H28	0.2333	−0.2484	0.6601	0.063*
C29	0.20123 (9)	−0.2242 (2)	0.78475 (14)	0.0479 (5)
C30	0.16019 (10)	−0.1435 (3)	0.83352 (14)	0.0549 (5)
H30	0.1570	−0.1717	0.8954	0.066*
C31	0.12380 (9)	−0.0198 (3)	0.78898 (13)	0.0523 (5)
H31	0.0956	0.0358	0.8210	0.063*
N2	0.43193 (9)	−0.0867 (2)	0.40558 (13)	0.0579 (5)
N1	0.40301 (7)	−0.1068 (2)	0.54169 (11)	0.0479 (4)
N3	0.26493 (10)	0.0946 (3)	1.00319 (13)	0.0648 (5)
N4	0.09446 (7)	0.47182 (19)	0.63073 (10)	0.0400 (3)
N5	0.03384 (8)	0.6791 (2)	0.59943 (13)	0.0534 (4)
N6	0.24265 (9)	−0.3485 (2)	0.83428 (16)	0.0687 (5)
O1	0.37147 (7)	0.20963 (18)	0.61034 (9)	0.0507 (3)
O2	0.27864 (10)	−0.0119 (3)	1.06269 (13)	0.0946 (6)
O3	0.22527 (9)	0.1998 (3)	1.00893 (13)	0.0938 (6)
O4	0.03272 (6)	0.18148 (19)	0.67729 (10)	0.0539 (4)
O5	0.24127 (9)	−0.3696 (3)	0.91796 (15)	0.0950 (6)
O6	0.27855 (12)	−0.4200 (3)	0.79039 (16)	0.1221 (9)
Cl1	−0.06934 (2)	−0.05900 (6)	0.59702 (3)	0.04943 (14)
Cl2	0.45182 (3)	0.48449 (7)	0.70812 (4)	0.06535 (18)
H1O	0.3900 (11)	0.295 (3)	0.6344 (17)	0.070 (8)*
H4O	0.0087 (13)	0.104 (4)	0.6494 (19)	0.089 (9)*
H5N	0.0057 (11)	0.751 (3)	0.6048 (16)	0.066 (7)*
H1N	0.4328 (11)	−0.053 (3)	0.3500 (19)	0.074 (8)*

Atomic displacement parameters (Ų)

	U^11	U^22	U^33	U^12	U^13	U^23
C1	0.0584 (12)	0.0572 (12)	0.0503 (11)	−0.0082 (10)	0.0062 (10)	−0.0071 (10)
C2	0.0614 (13)	0.0430 (11)	0.0592 (12)	−0.0035 (10)	0.0172 (10)	−0.0037 (9)
C3	0.0621 (13)	0.0513 (12)	0.0632 (13)	−0.0119 (11)	0.0258 (11)	−0.0156 (10)
C4	0.0637 (13)	0.0487 (11)	0.0569 (12)	−0.0106 (10)	0.0283 (10)	−0.0072 (9)
C5	0.0427 (10)	0.0431 (10)	0.0405 (9)	−0.0015 (8)	0.0098 (8)	0.0008 (8)
C6	0.0442 (10)	0.0374 (9)	0.0401 (9)	−0.0025 (8)	0.0083 (7)	0.0001 (8)
C7	0.0696 (14)	0.0671 (14)	0.0581 (13)	0.0287 (12)	0.0216 (11)	0.0187 (11)
C8	0.0798 (15)	0.0686 (14)	0.0497 (12)	0.0176 (13)	0.0187 (11)	0.0222 (11)
C9	0.0514 (11)	0.0511 (11)	0.0421 (10)	−0.0128 (9)	0.0155 (8)	−0.0025 (8)
C10	0.0425 (10)	0.0614 (13)	0.0504 (11)	0.0039 (9)	0.0105 (8)	−0.0020 (9)
C11	0.0465 (10)	0.0561 (12)	0.0405 (9)	0.0055 (9)	0.0054 (8)	0.0050 (9)
C21	0.0542 (11)	0.0501 (11)	0.0445 (10)	0.0042 (10)	0.0109 (9)	−0.0032 (9)
C22	0.0596 (12)	0.0643 (13)	0.0438 (10)	0.0154 (11)	0.0188 (9)	0.0152 (9)
C23	0.0673 (14)	0.0616 (13)	0.0559 (12)	0.0114 (11)	0.0090 (10)	0.0208 (11)
C24	0.0407 (9)	0.0506 (11)	0.0353 (9)	0.0058 (8)	0.0040 (7)	0.0061 (8)
C25	0.0483 (10)	0.0493 (11)	0.0322 (8)	0.0074 (9)	0.0037 (7)	−0.0010 (8)
C26	0.0433 (10)	0.0415 (10)	0.0380 (9)	0.0008 (8)	0.0035 (7)	−0.0032 (8)
C27	0.0569 (11)	0.0495 (11)	0.0394 (9)	0.0022 (9)	0.0124 (8)	−0.0027 (8)
C28	0.0532 (11)	0.0471 (11)	0.0583 (12)	0.0073 (9)	0.0129 (9)	−0.0106 (9)
C29	0.0493 (11)	0.0355 (9)	0.0565 (11)	0.0030 (8)	−0.0024 (9)	−0.0017 (8)
C30	0.0629 (13)	0.0607 (13)	0.0417 (10)	0.0092 (11)	0.0086 (9)	0.0089 (9)
C31	0.0558 (12)	0.0624 (13)	0.0406 (10)	0.0173 (10)	0.0136 (9)	0.0036 (9)
N2	0.0707 (12)	0.0623 (12)	0.0428 (10)	−0.0188 (10)	0.0152 (9)	−0.0106 (9)
N1	0.0553 (10)	0.0420 (9)	0.0494 (9)	−0.0097 (8)	0.0181 (7)	−0.0088 (7)
N3	0.0672 (12)	0.0808 (14)	0.0500 (10)	−0.0270 (11)	0.0215 (9)	−0.0065 (10)
N4	0.0407 (8)	0.0435 (8)	0.0366 (7)	0.0038 (7)	0.0077 (6)	0.0037 (6)
N5	0.0500 (10)	0.0429 (9)	0.0669 (11)	0.0090 (8)	0.0057 (8)	−0.0026 (8)
N6	0.0731 (13)	0.0499 (11)	0.0786 (14)	0.0145 (10)	−0.0082 (11)	−0.0017 (10)
O1	0.0662 (9)	0.0461 (8)	0.0402 (7)	−0.0043 (7)	0.0081 (6)	0.0022 (6)
O2	0.1329 (17)	0.0959 (14)	0.0623 (11)	−0.0300 (13)	0.0411 (11)	0.0140 (10)
O3	0.0742 (12)	0.1387 (18)	0.0768 (12)	0.0064 (13)	0.0418 (10)	−0.0079 (12)
O4	0.0428 (7)	0.0556 (9)	0.0623 (9)	0.0022 (7)	0.0033 (6)	−0.0020 (7)
O5	0.0931 (14)	0.1020 (15)	0.0873 (14)	0.0298 (11)	0.0020 (11)	0.0380 (12)
O6	0.153 (2)	0.1093 (17)	0.0980 (15)	0.0890 (17)	−0.0069 (14)	−0.0156 (13)
Cl1	0.0551 (3)	0.0540 (3)	0.0388 (2)	0.0064 (2)	0.00474 (19)	−0.0020 (2)
Cl2	0.0797 (4)	0.0661 (4)	0.0524 (3)	−0.0229 (3)	0.0167 (3)	−0.0063 (2)

Geometric parameters (Å, º)

C1—N2	1.315 (3)	C22—N4	1.370 (2)
C1—N1	1.323 (3)	C22—H22	0.9300
C1—H1	0.9300	C23—N5	1.359 (3)
C2—C3	1.341 (3)	C23—H23	0.9300
C2—N1	1.372 (3)	C24—N4	1.462 (2)
C2—H2	0.9300	C24—C25	1.518 (3)
C3—N2	1.358 (3)	C24—H24A	0.9700
C3—H3	0.9300	C24—H24B	0.9700
C4—N1	1.470 (2)	C25—O4	1.419 (2)
C4—C5	1.519 (3)	C25—C26	1.512 (3)
C4—H4A	0.9700	C25—H25	0.9800
C4—H4B	0.9700	C26—C27	1.384 (3)
C5—O1	1.412 (2)	C26—C31	1.388 (3)
C5—C6	1.523 (2)	C27—C28	1.380 (3)
C5—H5	0.9800	C27—H27	0.9300
C6—C11	1.377 (3)	C28—C29	1.374 (3)
C6—C7	1.380 (3)	C28—H28	0.9300
C7—C8	1.381 (3)	C29—C30	1.371 (3)
C7—H7	0.9300	C29—N6	1.469 (3)
C8—C9	1.367 (3)	C30—C31	1.378 (3)
C8—H8	0.9300	C30—H30	0.9300
C9—C10	1.365 (3)	C31—H31	0.9300
C9—N3	1.467 (3)	N2—H1N	0.85 (3)
C10—C11	1.383 (3)	N3—O2	1.219 (3)
C10—H10	0.9300	N3—O3	1.221 (3)
C11—H11	0.9300	N5—H5N	0.85 (2)
C21—N5	1.310 (3)	N6—O6	1.213 (3)
C21—N4	1.317 (2)	N6—O5	1.219 (3)
C21—H21	0.9300	O1—H1O	0.84 (3)
C22—C23	1.336 (3)	O4—H4O	0.87 (3)
N2—C1—N1	108.3 (2)	N4—C24—H24A	109.5
N2—C1—H1	125.9	C25—C24—H24A	109.5
N1—C1—H1	125.9	N4—C24—H24B	109.5
C3—C2—N1	107.0 (2)	C25—C24—H24B	109.5
C3—C2—H2	126.5	H24A—C24—H24B	108.0
N1—C2—H2	126.5	O4—C25—C26	113.14 (15)
C2—C3—N2	107.0 (2)	O4—C25—C24	106.57 (15)
C2—C3—H3	126.5	C26—C25—C24	109.22 (14)
N2—C3—H3	126.5	O4—C25—H25	109.3
N1—C4—C5	111.31 (15)	C26—C25—H25	109.3
N1—C4—H4A	109.4	C24—C25—H25	109.3
C5—C4—H4A	109.4	C27—C26—C31	119.12 (17)
N1—C4—H4B	109.4	C27—C26—C25	120.34 (16)
C5—C4—H4B	109.4	C31—C26—C25	120.49 (16)
H4A—C4—H4B	108.0	C28—C27—C26	120.91 (18)
O1—C5—C4	106.19 (15)	C28—C27—H27	119.5
O1—C5—C6	113.14 (15)	C26—C27—H27	119.5
C4—C5—C6	109.70 (15)	C29—C28—C27	118.30 (18)
O1—C5—H5	109.2	C29—C28—H28	120.8
C4—C5—H5	109.2	C27—C28—H28	120.8
C6—C5—H5	109.2	C30—C29—C28	122.34 (18)
C11—C6—C7	118.95 (17)	C30—C29—N6	118.47 (19)
C11—C6—C5	121.78 (16)	C28—C29—N6	119.16 (19)
C7—C6—C5	119.27 (17)	C29—C30—C31	118.69 (18)
C6—C7—C8	121.0 (2)	C29—C30—H30	120.7
C6—C7—H7	119.5	C31—C30—H30	120.7
C8—C7—H7	119.5	C30—C31—C26	120.57 (18)
C9—C8—C7	118.41 (19)	C30—C31—H31	119.7
C9—C8—H8	120.8	C26—C31—H31	119.7
C7—C8—H8	120.8	C1—N2—C3	109.37 (19)
C10—C9—C8	122.08 (18)	C1—N2—H1N	123.1 (18)
C10—C9—N3	118.76 (19)	C3—N2—H1N	127.5 (17)
C8—C9—N3	119.16 (19)	C1—N1—C2	108.38 (17)
C9—C10—C11	118.82 (18)	C1—N1—C4	125.60 (19)
C9—C10—H10	120.6	C2—N1—C4	126.00 (18)
C11—C10—H10	120.6	O2—N3—O3	123.4 (2)
C6—C11—C10	120.63 (18)	O2—N3—C9	118.3 (2)
C6—C11—H11	119.7	O3—N3—C9	118.3 (2)
C10—C11—H11	119.7	C21—N4—C22	108.02 (16)
N5—C21—N4	109.00 (17)	C21—N4—C24	126.17 (15)
N5—C21—H21	125.5	C22—N4—C24	125.63 (15)
N4—C21—H21	125.5	C21—N5—C23	108.80 (18)
C23—C22—N4	107.08 (18)	C21—N5—H5N	124.9 (16)
C23—C22—H22	126.5	C23—N5—H5N	126.1 (16)
N4—C22—H22	126.5	O6—N6—O5	123.2 (2)
C22—C23—N5	107.09 (18)	O6—N6—C29	118.0 (2)
C22—C23—H23	126.5	O5—N6—C29	118.7 (2)
N5—C23—H23	126.5	C5—O1—H1O	105.9 (17)
N4—C24—C25	110.93 (14)	C25—O4—H4O	110.0 (18)
N1—C2—C3—N2	0.3 (2)	C28—C29—C30—C31	−1.9 (3)
N1—C4—C5—O1	63.6 (2)	N6—C29—C30—C31	176.16 (19)
N1—C4—C5—C6	−173.80 (16)	C29—C30—C31—C26	−0.3 (3)
O1—C5—C6—C11	20.0 (2)	C27—C26—C31—C30	2.2 (3)
C4—C5—C6—C11	−98.3 (2)	C25—C26—C31—C30	−175.12 (19)
O1—C5—C6—C7	−159.94 (19)	N1—C1—N2—C3	−0.5 (2)
C4—C5—C6—C7	81.7 (2)	C2—C3—N2—C1	0.1 (2)
C11—C6—C7—C8	2.6 (3)	N2—C1—N1—C2	0.7 (2)
C5—C6—C7—C8	−177.4 (2)	N2—C1—N1—C4	179.03 (17)
C6—C7—C8—C9	−0.6 (4)	C3—C2—N1—C1	−0.6 (2)
C7—C8—C9—C10	−2.0 (4)	C3—C2—N1—C4	−178.94 (17)
C7—C8—C9—N3	178.1 (2)	C5—C4—N1—C1	−82.8 (2)
C8—C9—C10—C11	2.6 (3)	C5—C4—N1—C2	95.3 (2)
N3—C9—C10—C11	−177.49 (18)	C10—C9—N3—O2	165.1 (2)
C7—C6—C11—C10	−2.0 (3)	C8—C9—N3—O2	−14.9 (3)
C5—C6—C11—C10	178.03 (18)	C10—C9—N3—O3	−14.4 (3)
C9—C10—C11—C6	−0.5 (3)	C8—C9—N3—O3	165.6 (2)
N4—C22—C23—N5	−0.4 (3)	N5—C21—N4—C22	−0.9 (2)
N4—C24—C25—O4	−61.50 (17)	N5—C21—N4—C24	−176.16 (17)
N4—C24—C25—C26	175.96 (14)	C23—C22—N4—C21	0.8 (2)
O4—C25—C26—C27	145.34 (18)	C23—C22—N4—C24	176.07 (18)
C24—C25—C26—C27	−96.1 (2)	C25—C24—N4—C21	97.9 (2)
O4—C25—C26—C31	−37.4 (2)	C25—C24—N4—C22	−76.5 (2)
C24—C25—C26—C31	81.2 (2)	N4—C21—N5—C23	0.7 (2)
C31—C26—C27—C28	−2.0 (3)	C22—C23—N5—C21	−0.2 (3)
C25—C26—C27—C28	175.37 (18)	C30—C29—N6—O6	179.6 (2)
C26—C27—C28—C29	−0.2 (3)	C28—C29—N6—O6	−2.2 (3)
C27—C28—C29—C30	2.2 (3)	C30—C29—N6—O5	−3.6 (3)
C27—C28—C29—N6	−175.90 (18)	C28—C29—N6—O5	174.5 (2)