The crystal structures of the title spiro derivatives are described and the analysis of the intermolecular contacts in the crystals using Hirshfeld surface analysis and two-dimensional fingerprint plots is reported. a-hexahydrospiropyrrolothiazole-11,11′-indenoquinoxaline], C37H26N4O3S, (I), and 6′--6a′-nitro-6′,6a′,6b′,7′,8′,9′,10′,12a′-octahydro-2H-spiroindolizin]-2-one, C36H28N2O4, (II), are new spiro derivatives, in which both the pyrrolidine rings adopt twisted conformations. In (I), the five-membered thiazole ring adopts an envelope conformation, while the eight-membered pyrrolidine-thiazole ring adopts a boat conformation. An intramolecular C—H⋯N hydrogen bond occurs, involving a C atom of the pyran ring and an N atom of the pyrazine ring. In (II), the six-membered piperidine ring adopts a chair conformation. An intramolecular C—H⋯O hydrogen bond occurs, involving a C atom of the pyrrolidine ring and the keto O atom. For both compounds, the crystal structure is stabilized by intermolecular C—H⋯O hydrogen bonds. In (I), the C—H⋯O hydrogen bonds link adjacent molecules, forming R22(16) loops propagating along the b-axis direction, while in (II) they form zigzag chains along the b-axis direction. In both compounds, C—H⋯π interactions help to consolidate the structure, but no significant π–π interactions with centroid–centroid distances of less than 4 Å are observed.The title compounds, 6--6a-nitro-6,6a,6 b,7,9,11 The eight-membered pyrrolidine-thiazole ring (S1/C24–C27/C1/C16/N3) adopts a boat conformation with a total puckering amplitude Q = 1.351 (2) Å and φ = 321.43 (8)°. The mean planes of the pyran and thiazole rings are inclined to each other by 77.5 (2)°. The mean plane of the pyrazine ring (N1/N2/C8/C9/C14/C15) forms a dihedral angle of 57.1 (2)° with the mean plane of the pyran ring, while it is almost perpendicular with respect to the mean plane of the pyrrolidine ring, forming an angle of 89.8 (2)°. The pyrazine ring is inclined by 51.9 (2), 1.9 (2) and 69.5 (2)° with respect to the mean planes of the thiazole and cyclopentene ring and the naphthalene ring system, respectively. An intramolecular C23–H23⋯N1 hydrogen bond is formed ed Fig. 1.q2 = 0.045 (2) Å, θ = 175.7 (2)° and φ = 22 (3)°. The dihedral angle between the acenaphthylene (C1–C12) and naphthalene (C27–C36) ring systems is 63.8 (6)°. Moreover, this moiety is inclined of 85.3 (1), 36.1 (1) and 89.4 (2) ° with respect to the mean planes of the pyrrolidine (N1/C12/C17–C19), pyran (O4/C18–C20/C25/C26) and piperidine (N1/C13–C17) rings, respectively. The keto atom O1 deviates from the mean plane of the acenaphthylene unit by 0.148 (1) Å. An intramolecular C17—H17⋯O1 hydrogen bond is present nt Fig. 2.b-axis direction. The loops are linked by C—H⋯S hydrogen bonds, forming layers parallel to the (101) plane; C—H⋯π interactions are present within the layers on Fig. 4. A C—H⋯πet al., 2007CrystalExplorer17  through white to blue . The red spots on the surface indicate the intermolecular contacts involved in hydrogen bonding.The Hirshfeld surfaces of (I)b), C⋯H/H⋯C , O⋯H/H⋯O , S⋯H/H⋯S  and N⋯H/H⋯N , followed by the C⋯C contacts . For (II)b), C⋯H/H⋯C , O⋯H/H⋯O , followed by the C⋯C contacts . In both compounds the H⋯H intermolecular contacts predominate.The fingerprint plots for the two compounds are given in Figs. 9et al., 2016H,6′H,6b′H-spiroindolizin]-2-one skeleton yielded five hits: namely 6-(4- methoxyphenyl)-6a-nitro-6,6a,6b,7,8,9,10,12a-octahydrospiro-indolizine-12,3-indolin]-2-one  to a solution of indenoquinoxalinone  and thiazolidine-4-carboxylic acid  in dry toluene, 0.302 g (1.0 mmol) of 2--3-nitro-2H-chromene were added under a nitrogen atmosphere.Compound (II) to a solution of acenaphthoquinone  and pipacolinic acid  in dry toluene,  of 2--3-nitro-2H-chromene were added under a nitrogen atmosphere.The solutions were refluxed for 18 h in a Dean–Stark apparatus to give the cycloadducts. After completion of the reactions as indicated by TLC, the solvent was evaporated under reduced pressure. The crude products obtained were purified by column chromatography using hexane/EtOAc (7:3) as eluent (yield 84%). Colourless block-like crystals of the title compounds, suitable for X-ray diffraction analysis, were obtained by slow evaporation of solutions in ethanol.Uiso(H) = 1.5Ueq(C-methyl) and 1.2Ueq for all other H atoms.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S205698901901291X/xi2018sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S205698901901291X/xi2018Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S205698901901291X/xi2018IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S205698901901291X/xi2018Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S205698901901291X/xi2018IIsup5.cmlSupporting information file. DOI:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The dihedral angle between the 4-bromophenyl rings is 51.56 (2)°. In the crystal, molecules are linked into a zigzag chain propagating along [001] by weak C—H⋯π interactions. The conformations of related bischalcones are surveyed and a Hirshfeld surface analysis is used to investigate and quantify the intermolecular contacts.In the title bischalcone, C E,4E)-1,5-diphenylpenta-1,4-dien-3-one] was first prepared by the base-catalyzed Aldol condensation of benzaldehyde and acetone —C=C—) moiety. Bischalcones have a number of uses including anti-inflammatory ; τ4 (C13—C12—C11—C10)] and the carbonyl double bond  °; τ4 = −6.2 (4)°] but the conformations of the olefinic double bonds are very different: one is in s-trans [τ2 = 160.7 (3)°] conformation and in s-cis [τ3 = −15.2 (4)°] conformation with the central C=O double bond. These torsions result in an overall twisted shape for (I)The asymmetric unit of (I))] Fig. 2. The 4-bvia C1—H1A⋯Cg1i . The C14—H14A⋯Cg2ii  interactions lead to a zigzag chain along the c-axis direction.No classical hydrogen bonding is possible in (I)1i Fig. 3a. The Cii Fig. 3b interet al., 2016E,4E)-1,5-diphenylpenta-1,4-dien-3-one as the main skeleton revealed the presence of 27 structures containing a similar bischalcone moiety to the title compound but with different substituents on the terminal phenyl rings. The different substituents (1R and 2R) together with the torsion angles of the penta-4,4-dien-3-one connecting bridge are compiled in Table 2s-cis or s-trans), the olefinic double bonds are close to coplanar with their attached phenyl rings as indicated by their τ1 and τ4 torsion angles, which fall in the range of 0.0–17.8°, except for the compounds AMEXUN and HUDLEY, which have somewhat larger τ1 and τ4 values of 22.5–27.4°. The olefinic double bonds for the symmetrical compounds are mostly in s-cis conformations with the carbonyl double bond (τ2/τ3 torsion angles of 0.1–21.9°). However, both the olefinic double bonds of compounds GOLGOD and GOLGOD02 are in s-trans conformations with the carbonyl double bond (τ2/τ3 = 152.2–153.4°). Furthermore, it may be noted that the symmetrical conformation at the penta-4,4-dien-3-one connection bridge is not affected by the different substituents at the 1R and 2R positions in EDUSEE, SAFZOO and XOHVUN. Most of the unsymmetrical compounds (one C=C—C=O bond s-cis and one s-trans) have τ1 and τ4 values of 0.5–17.2°, which indicates that the olefinic double bonds are close to coplanar to their attached phenyl ring. The outliers are MESXEQ and WIHBUL, which have τ1 and τ4 values of 18.2–51.8° and 21.4–51.8°, respectively. The torsion angles τ2 and τ3 for the unsymmetrical compounds, including (I)s-trans and s-cis conformations between the olefinic double bonds and the carbonyl double bond.A survey of the Cambridge Structural Database CrystalExplorer17.5  correspond to the C14—H14A⋯Cg2ii interaction. Even through the C1—H1A⋯Cg1i interaction is not visible in the dnorm surface mapping, this interaction can be seen as a unique pattern of a red ‘circle’ on the shape-index surface mapping . Besides the C—H⋯π interactions, the dnorm surface mapping indicated a short contact between atom O1 and C5 with a distance of 0.06 Å shorter than the sum of the van der Waals radii of O and C atoms . Together with this short contact, another weak C7—H7A⋯O1 interaction was also revealed as light spots on the dnorm surface .The Hirshfeld surfaces mapped with normalized contact distance rm Fig. 4a corresng Fig. 4b. Besidce Fig. 5b.de and di diagonal axes. The H⋯C/C⋯H contacts are the most populated contacts and contribute 34.1% to the total intermolecular contacts, followed by H⋯H (22.1%), H⋯Br/Br⋯H (20.4%) and H⋯O/O⋯H (9.2%) contacts . The H⋯H contacts appear in the central region of the fingerprint plots with de = di = 2.4 Å . With the presence of relatively larger bromine atoms in the structure, the H⋯Br/Br⋯H contacts appear as symmetrical broad wing at diagonal axes of de + di ≃ 3.0 Å . Two symmetric spikes in the fingerprint plots with a short spike at de + di ≃ 2.7 Å represent the H⋯O/O⋯H contacts , indicating the presence of the weak C7—H7A⋯O1 interaction. The percentage contributions for other contacts are less than 15% in the Hirshfeld surface mapping.As illustrated in Fig. 6ts Fig. 6. As the  Å Fig. 6b. The H Å Fig. 6c. With  Å Fig. 6d. Two sts Fig. 6e, indicA mixture of 4-bromobenzaldehyde  and acetone  dissolved in absolute ethanol (30 ml) was slowly added to an aqueous solution of potassium hydroxide (4.0 g in 20 ml water). The mixture was vigorously stirred at room temperature for two h and then 20 ml chilled water was added. The resulting yellow precipitate was recovered by vacuum filtration and washed with cold water (100 ml). The crude product was recrystallized from absolute ethanol solution as yellow blocks.-1,5-Bis(4-bromophenyl)penta-1,4-dien-3-one; pure yellow solid , m.p. 484 K; IR νmax 594, 687, 813, 979, 1066, 1181, 1320, 1398, 1480, 1581, 1643 cm−1, UV–Vis λmax. 227 and 317 nm, 1H NMR:δH  7.02 , 7.45 , 7.54 , 7.67 ; 13C NMR:δC  124.62, 125.69, 129.25, 131.74, 133.29, 141.83, 188.22; HRMS (ES): MH+, found: 392 C17H12Br2O+ requires: 391.92.Uiso(H) = 1.5Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989019006480/hb7821sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019006480/hb7821Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019006480/hb7821Isup3.cmlSupporting information file. DOI: 1914420CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In the compound, the acid and base molecules are linked by a short hydrogen bond [O⋯O = 2.4393 (15) Å], in which the H atom is disordered over two positions with equal occupancies.The title compound was analysed as a disordered structure over two states,  9H7.5NO·C7H3.5ClNO4, was analysed as a disordered structure over two states, viz. co-crystal and salt, accompanied by a keto–enol tautomerization in the base molecule. The co-crystal is 4-chloro-2-nitrobenzoic acid–quinolin-4(1H)-one (1/1), C7H4ClNO4·C9H7NO, and the salt is 4-hydroxyquinolinium 4-chloro-2-nitrobenzoate, C9H8NO+·C7H3ClNO4−. In the compound, the acid and base molecules are held together by a short hydrogen bond [O⋯O = 2.4393 (15) Å], in which the H atom is disordered over two positions with equal occupancies. In the crystal, the hydrogen-bonded acid–base units are linked by N—H⋯O and C—H⋯O hydrogen bonds, forming a tape structure along the a-axis direction. The tapes are stacked into a layer parallel to the ab plane via π–π interactions [centroid–centroid distances = 3.5504 (8)–3.9010 (11) Å]. The layers are further linked by another C—H⋯O hydrogen bond, forming a three-dimensional network. Hirshfeld surfaces for the title compound mapped over shape-index and dnorm were generated to visualize the intermolecular interactions.The title compound, C D—H⋯A hydrogen bonding  in chloro- and nitro-substituted benzoic acid–pyridine derivative systems, we have shown that several compounds, namely, three compounds of quinoline with 3-chloro-2-nitrobenzoic acid, 4-chloro-2-nitrobenzoic acid and 5-chloro-2-nitrobenzoic acid -quinolinone (1/1), and the salt, 4-hydroxyquinolinium 4-chloro-2-nitrobenzoate, accompanied by a keto–enol tautomerization in the base molecule. The C10—O5 bond length [1.2956 (18) Å] is intermediate between a C—O single bond [1.36 Å in phenol] and a C=O double bond [1.23 Å in ketones of the (Car)2—C=O type]  of the acid molecule and the quinoline ring system (N2/C8–C16) of the base are slightly inclined to each other by a dihedral angle of 10.27 (6)°, while the carboxy group (O1/C7/O2) is twisted by 38.66 (18) and 45.93 (18)°, respectively, with respect to the C1–C6 ring and the N2/C8–C16 ring system. The dihedral angle between the C1–C6 ring and the nitro group (O3/N1/O4) is 50.33 (19)°.i, C8—H8⋯O5i and C9—H9⋯O1i; symmetry code as in Table 1a axis  −x + y − z]. The layers are further linked by another C—H⋯O hydrogen bond –(3)]. The π–π interactions between the benzene rings of the acid molecules [Cg1⋯Cg1iv] and between the quinoline ring systems of the base molecules  are indicated by blue and red triangles on the shape-index surfaces [arrows (4) and (5)].In order to visualize the intermolecular interactions, Hirshfeld surfaces for the acid and base molecules of the title compound, mapped over shape-index and i Table 1 are repret al., 2016H)-quinolinone (keto tautomer) showed one structure, namely, 4-amino-1-(2-(hydroxymethyl)-1,3-oxathiolan-5-yl)-2(1H)-pyrimidinone 4(1H)-quinolinone -quinolinone itself was reported by Nasiri et al.  of 4-hydroxyquinoline (0.075 g) with 4-chloro-2-nitrobenzoic acid (0.106 g) in a 1:1 molar ratio at room temperature.A (O1 site) and H1B (O5 site), respectively, with bond restraints of O—H = 0.84 (1) Å and with Uiso(H) = 1.5Ueq(O). In the final refinement, the occupancies were fixed at 0.5, and one outlier  was omitted. The N-bound H atom was refined freely [refined distance: N2—H2 = 0.89 (2) Å]. Other H atoms were positioned geometrically (C—H = 0.95 Å) and treated as riding, with Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S205698901901497X/lh5935sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S205698901901497X/lh5935Isup2.hklStructure factors: contains datablock(s) I. DOI: 1963942, 1963942CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The chains are linked into layers parallel to the bc plane by sets of four C—H⋯O hydrogen bonds and the layers are tied together by complementary π-stacking interactions.The dihydroqinoxalinone portion of the molecule is planar to within 0.0512 (12) Å. In the crystal, a combination of C—H⋯O and C—H⋯N hydrogen bonds together with slipped π-stacking and C—H⋯π(ring) interactions lead to the formation of chains extending along the  16H17N5O3, is build up from two fused six-membered rings linked to a 1,2,3-triazole ring, which is attached to an ethyl azido-acetate group. The dihydroqinoxalinone portion is planar to within 0.0512 (12) Å and is oriented at a dihedral angle of 87.83 (5)° with respect to the pendant triazole ring. In the crystal, a combination of intermolecular C—H⋯O and C—H⋯N hydrogen bonds together with slipped π-stacking [centroid–centroid distance = 3.7772 (12) Å] and C—H⋯π (ring) interactions lead to the formation of chains extending along the c-axis direction. Additional C—H⋯O hydrogen bonds link these chains into layers parallel to the bc plane and the layers are tied together by complementary π-stacking [centroid–centroid distance = 3.5444 (12) Å] interactions. The Hirshfeld surface analysis of the crystal structure indicates that the most important contributions for the crystal packing are from H⋯H (44.5%), H⋯O/O⋯H (18.8%), H⋯N/N⋯H (17.0%) and H⋯C/C⋯H (10.4%) interactions.The molecule of the title compound, C The pendant triazole ring is inclined to this plane by 87.83 (5)°.Dhyqnx⋯OEthazac, C—HEthazac⋯ODhyqnx, C5—HDhyqnx⋯NEthazac and C—HTrz⋯NDhyqnx  hydrogen bonds  rings [centroid–centroid distance = 3.7772 (12) Å] and by complementary C—HDhyqnx⋯Cg3 interactions [Cg3 is the centroid of the benzene ring B (C1–C6)] (Table 1bc plane by sets of four C—HDhyqnx⋯OEthazac hydrogen bonds (Table 1a-axis direction by inversion-related slipped π-stacking interactions between the A and B rings [centroid–centroid distance = 3.5444 (12) Å]  analysis  contacts -one (0.65 mmol) in ethanol (20 mL) was added ethyl azidoacetate (1.04 mmol). The mixture was stirred under reflux for 24 h. After completion of the reaction (monitored by TLC), the solution was concentrated and the residue was purified by column chromatography on silica gel by using as eluent a hexane/ethyl acetate (9/1) mixture. Crystals were obtained when the solvent was allowed to evaporate. The solid product isolated was recrystallized from ethanol to afford yellow crystals in 75% yield.To a solution of 3-methyl-1-(prop-2-ynyl)-3,4-dihydroquinoxalin-2 global, I. DOI: 10.1107/S2056989018014561/xu5945Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018014561/xu5945Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018014561/xu5945Isup4.cmlSupporting information file. DOI: 1873385CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In each of the title compounds, the packing is driven by C—H⋯F intertactions, along with a variety of C—H⋯O, C—O⋯π, and C—F⋯π contacts. Hirshfeld surface analyses were conducted to aid in the visualization of these various influences on the packing. 14H8F4N2O2 and C14H8F4N2O3, are reported. In each crystal, the packing is driven by C—H⋯F intertactions, along with a variety of C—H⋯O, C—O⋯π, and C—F⋯π contacts. Hirshfeld surface analysis was conducted to aid in the visualization of these various influences on the packing: they showed that the largest contributions to the surface contacts arise from H⋯F/F⋯H interactions, followed by H⋯H and O⋯H/H⋯O.The syntheses and crystal structures of the title compounds, C In molecule A, the N1/C1–C5 plane is rotated by 58.05 (5)° relative to the N2/C6/C7/C12/C13 plane and the corresponding dihedral angle for molecule B is 61.65 (7)°. The addition of an oxygen atom between N2 and C3 in II alleviates this steric restriction and only one 19F NMR peak in solution is observed for the ortho-F atoms; even so, the dihedral angle between the N1/C1–C5 and N2/C6/C7/C12/C13 planes in the crystal of II of 84.01 (5)° is larger than those found in I.Compound it Fig. 1. The synI and II are of the type C—H⋯O, C—H⋯F, C—O⋯π, and C—F⋯π  and the olefinic hydrogen atoms . A weak interaction is also observed for a bridge hydrogen atom in II, C14—H14B⋯F4iii. The packing is further aided by π-interactions with the pyridine ring .The main directional interactions in the crystal structures of  Tables 1 and 2 ▸.CrystalExplorer17.5  to 1.3800 (blue) a.u. The pale-red spots symbolize short contacts and negative dnorm values on the corresponding surface plots shown in Fig. 2Hirshfeld surface analysis  < 1.25 Å in I and 1.10 Å < (di + de) < 1.35 Å in II. H⋯H contacts make the second largest contribution (20.2% in I and 14.1% in II), shown in the middle region 1.10 Å < (di + de) < 1.18 Å in I and II. The third largest contribution is from O⋯H/H⋯O contacts. In I, the corresponding spike is partially overlapped with the spike representing F⋯H/H⋯F contacts, appearing at 1.05 Å < (di + de) < 1.40 Å. The O⋯H/H⋯O spike is clearly visible in the fingerprint plot of II, shown in the region of 1.10 Å < (di + de) < 1.40 Å.The largest contribution to the overall crystal packing in both compounds is from F⋯H/H⋯F interactions -dione-based compounds with an aromatic substituent on the nitrogen atom yielded 58 results. The dihedral angle between the aromatic ring plane and the succinimide plane is bimodally distributed between 43 and 90°, with peaks near 60 and 75°.A search of the November 2018 release of the Cambridge Structure Database (Groom Synthesis of (I) pentafluoropyridine , -3a,4,7,7a-tetrahydro-1H-4,7-methanoisoindole-1,3(2H)-dione , and triethylamine  were combined in DMF (150 ml). The resulting solution was stirred at room temperature for 24 h. Diethyl ether (150 ml) and saturated aqueous ammonium chloride (100 ml) were added and the biphasic solution stirred vigorously for 2 h. The organic layer was separated and the remaining aqueous portion extracted with diethyl ether (2 × 150 ml). The combined organic fractions were washed with water (2 × 1 l) and brine (2 × 300 ml), dried over MgSO4, and the solvent removed via rotary evaporation. The resulting off-white solid was dissolved in refluxing EtOH (20 ml) and cooled to 278 K for 12 h. Vacuum filtration, washing with cold EtOH (20 ml), and vacuum drying afforded the target compound as a white, crystalline solid . 1H NMR : 6.28 , 3.58 , 3.54 , 1.74 .19F NMR : −90.5 (2F), −141.7 (1F), −143.1 (1F).Synthesis of (II) to a stirred solution of potassium carbonate , -2-hydroxy-3a,4,7,7a-tetrahydro-1H-4,7-methanoisoindole-1,3(2H)-dione , pentafluoropyridine , and 140 ml of DMF were added. The resulting solution was stirred at room temperature for 24 h. Diethyl ether (150 ml) and saturated aqueous ammonium chloride (100 ml) were added and the biphasic solution stirred vigorously for 2 h. The organic layer was separated and the remaining aqueous portion extracted with diethyl ether (2 × 150 ml). The combined organic fractions were washed with water (2 × 1 l) and brine (2 × 300 ml), dried over MgSO4, and the solvent removed via rotary evaporation. The resulting off-white solid was dissolved in refluxing EtOH (50 ml) and cooled to 278 K for 12 h. Vacuum filtration, washing with cold EtOH (20 ml) and vacuum drying afforded the target compound as a white, crystalline solid . 1H NMR : 6.23 , 3.50 , 3.34 , 1.68 .19F NMR : −87.4 (2F), −156.3 (2F).Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989019009769/hb7832sup1.cifCrystal structure: contains datablock(s) global, I, II. DOI: 10.1107/S2056989019009769/hb7832Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989019009769/hb7832IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989019009769/hb7832Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989019009769/hb7832IIsup5.cmlSupporting information file. DOI: 1879244, 1885019CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The title compound crystallizes with two independent molecules in the asymmetric unit. In the crystal, molecules are linked by N—H⋯O hydrogen bonds, C—H⋯π, I⋯S and I⋯I interactions into a three-dimensional network. A and B) are present in the asymmetric unit of the title compound, C11H9IN2OS, which differ mainly in the dihedral angle between the phenyl and thiazole rings . In the crystal, the molecules form ⋯A⋯B⋯A⋯B⋯ chains along the [001] and [010] directions through moderate N—H⋯O hydrogen bonds and C—H⋯π interactions, respectively. The overall three-dimensional network is formed by I⋯I and I⋯S interactions. Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from H⋯C/C⋯H (26.2%), H⋯H (20.9%), H⋯I/I⋯H (19.4%) and H⋯O/O⋯H (6.8%) interactions.Two crystallographically independent molecules ( Unlike the related compound 2-acetamido-4-p-tolyl-1,3-thiazole .The title 2-acetoamidothiazole derivative crystallizes in the monoclinic space group it Fig. 1. The privia a C(4) synthon  contacts [3.7758 (9) Å]  contacts of type I [θ1 = θ2 = 146.91 (8)°] with a length of 3.8547 (5) Å.In the crystal, molecules are linked by N1—H1⋯O2 and N3—H3⋯O1 moderate hydrogen bonds n Table 1, formingÅ] Fig. 3. AdditioCrystal Explorer17.5  to 1.3937 (blue) a.u. . The pair of characteristic wings in this fingerprint plot corresponds to the C—H⋯π interactions between the phenyl groups . The H⋯H and H⋯I/I⋯H contacts  make similar contributions to the total Hirshfeld surface of 20.9 and 19.4%, respectively. The reciprocal H⋯O/O⋯H interactions (6.8%) are seen as sharp symmetrical spikes with tips at ed + id ∼1.9 Å and arising from the N—H⋯O hydrogen bond . Intermolecular I⋯S/S⋯I  and I⋯I interactions make smaller contributions to the Hirshfeld surface .A Hirshfeld surface analysis was carried out using u. Fig. 4. The intps Fig. 5b. The Hts Fig. 5c and 5dnd Fig. 5e. Inter⋯I Fig. 5f and I⋯et al., 2016N--4-chlorobutanamide -1,3-thiazol-2-yl)-4-chlorobutanamide. In this compound the dihedral angle is smaller [8.8 (3)°] as a result of an intramolecular C—H⋯Br hydrogen bond. In the crystals of these compounds, only 5,5′-dibromo-4,4′-bis(pentafluorophenyl)-2,2′-bi-1,3-thiazole exhibits a type II halogen–halogen interaction with a Br⋯Br distance of 3.6777 (3) Å and angles of 68.88 (5) and 174.77 (5)°.A search of the Cambridge Structural Database  acetamide  and iodine  was placed in an open vessel containing a Teflon-coated stir bar. The mixture was dissolved in 3 mL of ethanol and the vessel was placed in the microwave cavity  and subjected to MW irradiation (150 W) for 60 min, at 363 K and a pressure of 2 psi. The reaction mixture was then cooled at room temperature and 5 mL of NH4OH were added. The obtained mixture was dissolved in ethyl acetate (50 mL) and washed with brine (3×). The organic layer was separated, dehydrated with Na2SO4, and evaporated in vacuo until dryness. The product was purified by flash column chromatography  with a mixture of petrol–dichloromethane–acetone (5:3:2). The title compound was obtained as pale-yellow needles in 30% yield . A diluted solution of the compound was prepared in hexane and kept on a dry and dark place at room temperature. Crystals were obtained after one week of slow evaporation. Spectroscopic data: 1H NMR : 11.37 , 7.80 , 7.43 , 1.62 . 13C NMR : 168.8 (s), 163.6 (s), 151.4 (s), 134.5 (s), 129.0 (d), 128.9 (d), 128.7 (d), 62.4 (s) 21.9 (c).A mixture of Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-methyl).Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989019004791/lh5897sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019004791/lh5897Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019004791/lh5897Isup3.cmlSupporting information file. DOI: 1908908CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


I (HypSiBr), and II (TipSiBr), crystallized in the cubic space group PaPThe title compounds,  9H27BrSi4  and C27H63BrSi4 , are described. Compound I was prepared in 85% yield by free-radical bromination of 1,1,1,3,3,3-hexamethyl-2-(trimethylsilyl)trisilane using bromobutane and 2,2′-azobis(2-methylpropionitrile) as a radical initiator at 333 K. The molecule possesses threefold rotational symmetry, with the central Si atom and the Br atom being located on the threefold rotation axis. The Si—Br bond distance is 2.2990 (12) Å and the Si—Si bond lengths are 2.3477 (8) Å. The Br—Si—Si bond angles are 104.83 (3)° and the Si—Si—Si bond angles are 113.69 (2)°, reflecting the steric hindrance inherent in the three trimethylsilyl groups attached to the central Si atom. Compound II was prepared in 55% yield by free-radical bromination of 1,1,1,3,3,3-hexaisopropyl-2-(triisopropylsilyl)trisilane using N-bromosuccinimide and 2,2′-azobis(2-methylpropionitrile) as a radical initiator at 353 K. Here the Si—Br bond length is 2.3185 (7) Å and the Si—Si bond lengths range from 2.443 (1) to 2.4628 (9) Å. The Br—Si—Si bond angles range from 98.44 (3) to 103.77 (3)°, indicating steric hindrance between the three triisopropylsilyl groups.The synthesis and crystal structures of two trissilyl bromide compounds, C The Si1—Br1 bond length is 2.2990 (12) Å. As for Si2, the C—Si2—C bond angles range from 107.1 (2) to 110.55 (17)°, while the C—Si2—Si1 bond angles range from 108.61 (10) to 110.16 (11)°.The molecular structure of compound II (TipSiBr), is shown in Fig. 2P4 descriptor for fourfold coordination of 0.90. The Br1—Si1—Si2/Si3/Si4 bond angles range from 98.44 (3) to 103.77 (3)°, and the Si1—Br1 bond distance is 2.3185 (7) Å, which is longer than that of compound I [2.2990 (12) Å]. The τ4 descriptor values for atoms Si2, Si3 and Si4 (the silicon atoms of the triisopropylsilyl groups) are 0.96, 0.97 and 0.95, respectively, indicating that their coordination geometry is closest to an ideal tetrahedron.The asymmetric unit of compound I or II. Compound II, however, contains four intramolecular C—H⋯Br hydrogen bonds (Table 2D⋯A distances that range from from 3.584 (3) to 3.726 (3) Å, and D—H⋯A bond angles that range from 131 to 155°.There are no significant intermolecular contacts, other than weak van der Waals interactions, present in the crystals of compounds s Table 2. These het al., 20163Si group. Of these, there are only 42 structures where the central silicon atom is bonded directly to a halogen.The Cambridge Structural Database  . The protio derivative (HypSiH) is a liquid at room temperature, and the structure of the iodo derivative (HypSiI) has not been deposited in the CSD.For compounds IV (TipSiH) was not found in the CSD, but the journal article 3SiBr, viz. compound II (TipSiBr). Like compounds I and III, compound IV crystallizes in the cubic space group PaiPr3)Si– groups to push further away from one another, resulting in Si2—Si1—Si2i,ii bond angles of 117.9 (1)° and Si2i,ii—Si1—H bond angles of 98.3 (1)° . In II, the corresponding Si—Si—Si bond angles range from 115.02 (4) to 116.59 (4)° and the Si—Si—Br bond angles vary from 98.44 (3) to 103.77 (3)°.The X-ray data for compound Compound I: Tris(trimethylsilyl)silane  was added to an oven-dried nitrogen-flushed 250 ml Schlenk flask with a stir-bar. Bromobutane  was filtered through a plug of silica gel in a Pasteur pipette and was transferred into the Schlenk flask. AIBN  was then added to the flask, and the reaction was heated to 333 K using an oil bath and then heating was stopped. After stirring the reaction overnight at room temperature, GC–MS analysis of a sample indicated incomplete reaction and more AIBN (11 mg) was added to the flask. The reaction was heated once more to 333 K for 1 h. Analysis by GC–MS now indicated that the reaction was complete. The flask was placed in a freezer at 243 K and colourless block-like crystals of I formed overnight. Removal of the solvent in vacuo yielded 2.2 g (85%). 1H NMR  δ 0.24 ; 13C NMR  δ −0.51 ppm; GC–MS: 11.24 min, m/z = 328, base peak: 73.Compound II: Tris(triisopropylsilyl)silane  was dissolved in freshly distilled benzene (10 ml) along with NBS (45 mg) and AIBN . The mixture was heated using an oil bath at 333 K for 30 min, when GC–MS analysis indicated that no reaction had occurred. At this point the solution was heated with a heat gun until the reaction mixture turned slightly yellow. The yellow colour dissipated in less than 1 min. Analysis of the reaction mixture by 1H NMR indicated that only 60% of the starting material had been consumed. An additional amount of NBS  was added to the reaction flask, and the solution was again heated with a heat gun. The product was isolated by removing the solvent in vacuo and extracting the product from the crude reaction mixture with pentane. The pentane solution was filtered through glass wool, concentrated and weighed (135 mg). Analysis of the product with 1H NMR indicated this was 90% pure. The product was further purified by dissolving this solid in 1 ml pentane, cooling to 195 K and isolating the colourless needle-like crystals of II by removing the solvent with a syringe, washing with pentane and drying in vacuo . 1H NMR  δ 1.34 , 1.66 ; 13C NMR  δ 16.4, 21.6; HRMS for C17H63BrSi4 calculated 535.2642 (M − C3H7), found 535.2641.Uiso(H) = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989018009696/su5451sup1.cifCrystal structure: contains datablock(s) I, II, Global. DOI: 10.1107/S2056989018009696/su5451Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989018009696/su5451IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989018009696/su5451Isup4.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018009696/su5451IIsup5.cmlSupporting information file. DOI: 1854536, 1854535CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The title compound comprises four crystallographically different molecules that are composed of a 1,5-bis(4-methoxyphenyl)penta-1,4-dien-3-ylidenyl group and a benzyl ring connected by a hydrazine-1-carbodithioate bridge. In the crystal, molecules are connected into a three-dimensional network through C—H⋯O, N—H⋯S and C—H⋯π interactions. 27H26N2O2S2, the asymmetric unit is comprised of four molecules (Z = 8 and Z′ = 4). The 4-methoxyphenyl rings are slightly twisted away from their attached olefinic double bonds [torsion angles = 5.9 (4)–19.6 (4)°]. The azomethine double bond has an s-trans configuration relative to one of the C=C bonds and an s-cis configuration relative to the other . The torsion angles between the azomethine C=N double bond and hydrazine-1-carbodithioate moiety indicate only small deviations from planarity, with torsion angles ranging from 0.9 (3) to 6.9 (3)° and from 174.9 (3) to 179.7 (2)°, respectively. The benzyl ring and the methylenesulfanyl moiety are almost perpendicular to each other, as indicated by their torsion angles [range 93.7 (3)–114.6 (2)°]. In the crystal, molecules are linked by C—H⋯O, N—H⋯S and C—H⋯π(ring) hydrogen-bonding interactions into a three-dimensional network. Structural details of related benzyl hydrazine-1-carbodithioate are surveyed and compared with those of the title compound.In the title hydrazinecarbodithioate derivative, C S-benzyl and S-alkyl dithiocarbazates are interesting ligands in coordination chemistry because they can act as N,S-chelating agents because of the presence of soft sulfur and hard nitrogen donor atoms  than their transition-metal complexes . Structural details of (I) are compared with other hydrazinecarbodithioates.Encouraged by previous findings on various properties of related Schiff base derivatives, we report herein the synthesis and structure determination of the title compound (I) consists of four molecules, denoted as A, B, C and D. Orientational disorder of the 1,5-bis(phenyl)penta-1,4-dien-3-ylidenyl amine moiety in molecule B and of the phenylmethyl moiety in molecule D were observed. The molecules are composed of a 1,5-bis(4-methoxyphenyl)penta-1,4-dien-3-ylidenyl moiety, connected to a benzyl ring by a hydrazine-1-carbodithioate (–C=N—(NH)—(C=S)—S—C–) bridge ; τ6 (C16—C15—O2—C27)], between the 4-methoxyphenyl ring and the olefinic double bond , and between the olefinic double bond and the azomethine double bond . Torsion angles τ1 and τ6 are approximately 0° or ±180° in the majority of the four molecules, except molecule C which has a τ1 of 20.9 (3)° (Table 1τ2 = 5.9 (4)–16.4 (13)°; τ5 = 7.5 (3)–19.6 (4)°]. The orientations of the the azomethine double bond with its neighbouring olefinic double bonds relative to the intermediate C—C bond  are different: one is in s-trans [τ3 = 147.4 (6)–175.7 (2)°] conformation and the other in s-cis [τ4 = 15.3 (3)–37.4 (7)°] conformation. The dihedral angles between two 4-methoxyphenyl rings in an individual molecule are in the range 23.59 (12)–89.6 (5)° (Table 2τ7 (C9—N1—N2—C18), τ8 (N1—N2—C18—S1), τ9 (N2—C18—S1—C19), τ10 (C18—S1—C19—C20) and τ11 (S1—C19—C20—C21). In all molecules of (I), the hydrazine-1-carbothioate bridges are more or less planar –6.9 (3)° and 174.9 (3)–179.7 (2)°, respectively). The torsion angles between the sulfane moiety and the methylene moiety indicate a slight twist [τ10 = 160.53 (17)–163.36 (16)°]. These contortions are more severe between the benzyl ring and the methylene sulfane moiety where τ11 is considerably smaller [τ11 = 93.7 (3)–114.6 (2)°]. The dihedral angles between the benzyl ring and the two 4-methoxyphenyl rings are in the range 31.6 (5)–89.9 (8)° . In addition, molecule C and molecule D are connected through C17C—H17C⋯O1D hydrogen bonds . The four molecules are linked into an endless chain parallel to [021] through the combination of these hydrogen bonds hydrazine-1-carbodithioate as reference moiety resulted in 45 structures with different substituents. The reference moiety and relevant torsion angles are illustrated in Fig. 61R) together with the torsion angles for the benzyl hydrazine-1-carbothioate moiety in these structures are collated in Table 4τ7, τ8 and τ9; torsion angles τ7 and τ9 range from 165.1 to 180.0° and indicate an anti-periplanar conformation whereas torsion angle τ8 is indicative of a syn-periplanar conformation (0.0– 9.1°). With respect to torsion angle τ10, most of the structures adopt an anti-periplanar conformation ranging from 159.5 to 180.0°, but there are nine structures that adopt either a syn-clinal or an anti-clinal conformation (77.6–110.4°). In most of the structures, the benzyl ring and the methylene sulfane moiety are orientated almost perpendicular to each other, as indicated by torsion angle τ11. Here, either a syn-clinal (68.1–88.4°) or an anti-clinal (92.2–141.1°) conformation is adopted. However, there is one outlier  was synthesized following a well-described literature protocol  was dissolved in absolute ethanol (50 ml) under heating and stirring. The resulting solution was slowly added to a hot solution of di-p-methoxybenzalacetone  dissolved in absolute ethanol (50 ml). 3-5 drops of concentrated hydrochloric acid were added to the mixture, which was subsequently heated and stirred for 5 h : 3145, ν(N—H); 1630, ν(C=N); 1244, ν(N—N); 1059, ν(C=S); 3145. 1H NMR (CDCl3) δ (p.p.m.)= 6.55–7.68 (aromatic H); 3.85 (OCH3); 4.55 (–CH2 benzyl); 10.20 (N—H). 13C NMR (CDCl3) δ(p.p.m.)= 114.35–142.88 (aromatic C); 55.55 (OCH3); 39.34 (–CH2 benzyl); 161.22 (C=N); 206.91 (C=S). m/z calculated for C27H26N2O2S2: 474, found 474.Yield: 57.3%. m.p.: 375-376 K. Analysis calculated for CA, B, C or D. The N-bound H atoms were located in difference-Fourier maps and were refined freely [N—H = 0.81 (3)–0.91 (4) Å]. The C-bound H atoms were positioned geometrically (C—H = 0.93–0.97 Å) and refined using a riding model, with Uiso(H) = 1.2 or 1.5Ueq(C). A rotating-group model was applied to the methyl groups. The 1,5-bis(phenyl)penta-1,4-dien-3-ylidenyl amine moiety in molecule B and the phenylmethyl moiety in molecule D display positional disorder, with refined site occupancy ratios of 0.667 (7):0.333 (7) and 0.653 (15):0.347 (15), respectively .Crystal data, data collection and structure refinement details are summarized in Table 510.1107/S2056989019013458/wm5520sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989019013458/wm5520Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989019013458/wm5520Isup3.cmlSupporting information file. DOI: 1902915, 1902915CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


A third monoclinic polymorph of the title compound is described. In the crystal, it exhibits C—H⋯Cl hydrogen bonds and face-on Cl⋯π interactions involving the chloroform disolvate molecules. Intermolecular weak offset π–π interactions are also present between the aromatic rings of the ligands. 2(C26H22P2)]·2CHCl3 (I), is the third monoclinic polymorph of this platinum(II) complex involving the bidentate ligand cis-1,2-bis(diphenylphosphanyl)ethylene (cis-dppe) , linking the molecules to form supramolecular sheets that lie in the bc plane.The title compound, [PtCll. 1998a. Inorg.  al. 1995. Inorg.  A selection of recent examples include complexes involving iron(II) (Song cis-dppe ligand and the title compound (I)II metal center with bidentate coordination by the phosphorus atoms of the cis-dppe ligand. The metal coordination sphere is completed by two chloride anions.The molecular structures of the et al., 1998aP21/n without solvent in the unit cell, while structure ZOLYII As for the previously reported polymorphs of compound (I)II center of all three structures are, unsurprisingly, quite similar. The Pt—P bond lengths range from 2.210 (2) to 2.219 (2) Å, while the Pt—Cl bond lengths range from 2.358 (2) to 2.366 (3) Å. The P—Pt—P bond angles range from 86.66 (11) to 87.08 (5)°, while the Cl—Pt—Cl bond angles range from 90.33 (7) to 91.03 (5)°. The τ4 descriptor for fourfold coordination  to 3.789 (10) Å, while the D—H⋯A bond angles range from 132 to 158°. Three face-on Cl⋯π interactions nd Fig. 2. The Cl⋯bc plane, as shown in Fig. 3Cg2⋯Cg2ii = 4.096 (6) Å , and Cg3⋯Cg4iii = 3.770 (6) Å [Cg3 and Cg4 are the centroids of rings C15–C20 and C21–C26, respectively, α = 5.3 (5)°, interplanar distances are 3.326 (4) and 3.439 (4) Å, slippage = 1.544 Å, symmetry code (iii) −x + 1, y − z + The complex molecules are also linked by weak offset π–π interactions, forming sheets that lie in the 2Cl2 solvent molecule and one CHCl3 solvent molecule in the unit cell, also shows Cl⋯π interactions. However, the methylene chloride solvent molecule is not engaged in a hydrogen bond with a chlorine atom of the PtII complex, and is disordered in the crystal lattice.The closely related polymorph ZOLYII, which contains one CHet al., 2016cis-dppe ligand is coordinated to a PtII center. In addition to the two polymorphs described above, the most similar cis-dppe–PtII coordination complexes include AFEXEO . Another structure related to the title compound is KADQEL  in a NMR tube. This solution was left to stand at room temperature, and colorless needle-like crystals of compound (I)The title compound was prepared serendipitously by mixing 20.5 mg of Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989018008836/su5444sup1.cifCrystal structure: contains datablock(s) Global, I. DOI: 10.1107/S2056989018008836/su5444Isup2.hklStructure factors: contains datablock(s) I. DOI: 1849747CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The synthesis of a hybrid molecule is reported. The crystal structure of the monohydrate was investigated using Hirshfeld surface analysis and enrichment contact ratios. Hydrogen bonds induced by guest water molecules are the main driving force in crystal packing formation. 14H14N2OS2·H2O, the planar imidazopyridine ring system is linked to the 1,3-dithiolane moiety by an enone bridge. The atoms of the C—C bond in the 1,3-dithiolane ring are disordered over two positions with occupancies of 0.579 (14) and 0.421 (14) and both disordered rings adopt a half-chair conformation. The oxygen atom of the enone bridge is involved in a weak intramolecular C—H⋯O hydrogen bond, which generates an S(6) graph-set motif. In the crystal, the hybrid molecules are associated in R22(14) dimeric units by weak C—H⋯O interactions. O—H⋯O hydrogen bonds link the water molecules, forming infinite self-assembled chains along the b-axis direction to which the dimers are connected via O—H⋯N hydrogen bonding. Analysis of intermolecular contacts using Hirshfeld surface analysis and contact enrichment ratio descriptors indicate that hydrogen bonds induced by water molecules are the main driving force in the crystal packing formation.In the title hydrated hybrid compound C Its geometrical parameters are similar to those found for 1--3,3-bis(methylsulfanyl)prop-2-enone  = 0.419 (7)/0.443 (9) Å, φ(2) = 303.2 (9)/128.9 (11)° for the major and minor components, respectively]. The oxygen atom of the linker moiety is involved in a weak intramolecular C6—H6⋯O1 hydrogen bond (Table 1S(6) graph-set motif.Fig. 1d Table 1, which gvia pairwise weak C—H⋯O interactions [H5⋯O1i = 2.71 Å; symmetry code as in Table 1W—H1W⋯O2Wii hydrogen bonds, generating an infinite self-assembled chain of water molecules in a helical fashion along the b axis around which the host molecules are linked via O2W—H2W⋯N1 hydrogen bonds and weak C12—H12D⋯O2Wii interactions .In the crystal, the host molecules form inversion dimers ns Fig. 4. The hoson Fig. 5 and eachet al., 2007CrystalExplorer  are displayed in Figs. 6W—H2W⋯N1 hydrogen bond while the pale-red spot near H12B illustrates the weak C—H⋯O2W interaction. The white spots represent H⋯O, H⋯S and H⋯H contacts. On the shape-index surface, convex blue regions indicate hydrogen-donor groups, while concave red regions indicate hydrogen-acceptor groups and S⋯N and S⋯C contacts and O⋯C interactions. The fingerprint plots show the contribution of different types of intermolecular interactions  is defined as the ratio between the proportion of actual crystal contacts between the different chemical species  and the theoretical proportion of random equiprobable contacts . The results obtained are summarized in Table 2ECHc = 0.76 and correspond to weak C—H⋯C interactions. These interactions are under-represented because competition with the S⋯Hc, OW⋯Hc and weak O⋯Hc hydrogen bonds, the first two of which appear favoured with enrichment values of 1.35 and 1.14, respectively, and the last slightly under-represented with an enrichment ratio of 0.98. The C⋯C contacts are privileged and display an enrichment value of 1.85, which highlight molecules stacking one on top of the other as shown in Fig. 5et al. -3,3-bis(methylsulfanyl)prop-2-∊none monohydrate ethanone (6.2 mmol) was dissolved in distilled dimethyl sulfoxide (15 ml), and the carbon disulfide  was added. After cooling the mixture to 273 K, sodium hydride  was added. After stirring for 30 min. at 273 K, the mixture was stirred at ambient temperature for 4 h. The solution was then cooled at 273 K and 1,2-dichloro ethane  was added dropwise. The resulting mixture was then stirred for 24 h and then poured into 50 ml of ice-cold water. The precipitate was filtered and recrystallized from a mixture of water–dioxane (2:1) to obtain brown single crystals of the title compound suitable for X-ray diffraction analysis .1- and were refined using a riding model with Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-methyl). In the 1,3-dithiolane ring, the carbon atoms of the C—C bond are disordered over two positions with refined occupancy factors of 0.579 (14) and 0.421 (14). C—C bond lengths in both disordered components were restrained to the target value of 1.513 Å  I. DOI: Click here for additional data file.10.1107/S2056989019015755/vm2224Isup2.cmlSupporting information file. DOI: 1967239CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


PTPN2 deletion in T cells enhances cancer immunosurveillance and the efficacy of adoptively transferred tumour‐specific T cells. T‐cell‐specific PTPN2 deficiency prevented tumours forming in aged mice heterozygous for the tumour suppressor p53. Adoptive transfer of PTPN2‐deficient CD8+ T cells markedly repressed tumour formation in mice bearing mammary tumours. Moreover, PTPN2 deletion in T cells expressing a chimeric antigen receptor (CAR) specific for the oncoprotein HER‐2 increased the activation of the Src family kinase LCK and cytokine‐induced STAT‐5 signalling, thereby enhancing both CAR T‐cell activation and homing to CXCL9/10‐expressing tumours to eradicate HER‐2+ mammary tumours in vivo. Our findings define PTPN2 as a target for bolstering T‐cell‐mediated anti‐tumour immunity and CAR T‐cell therapy against solid tumours.Although adoptive T‐cell therapy has shown remarkable clinical efficacy in haematological malignancies, its success in combating solid tumours has been limited. Here, we report that  Protein tyrosine phosphatase N2 inhibits tumour T‐cell infiltration and CAR T‐cell cytotoxicity and may thus be a therapeutic target to enhance solid‐tumour immunotherapy. Upon necropsy 15/28 (54%), Ptpn2fl/fl;p53+/− mice developed various tumours including thymomas, lymphomas, sarcomas, carcinomas and hepatomas  were evident in the thymi or peripheral lymphoid organs of 5/28 mice  developed any overt tumours, splenomegaly or abnormal lymphocytic populations as assessed by gross morphology or flow cytometry and lymphoid organ tissue architecture was normal ‐expressing AT‐3 (AT‐3‐OVA) mammary carcinoma cells implanted into the inguinal mammary fat pads of Ptpn2fl/fl versus Lck‐Cre;Ptpn2fl/fl C57BL/6 mice  T cells into tumours  were increased rather than decreased in AT‐3‐OVA tumours ‐γ and tumour necrosis factor (TNF)  peptide SIINFEKL. Naive OT‐1 T cells can undergo clonal expansion and develop effector function when they engage OVA‐expressing tumours, but thereon leave the tumour microenvironment, become tolerised and fail to control tumour growth  Ptpn2fl/fl OT‐1 CD8+ T cells had no overt effect on the growth of AT‐3‐OVA mammary tumours when compared to vehicle‐treated tumour‐bearing mice  by 25 days. Tumour re‐emergence in this setting was accompanied by decreased OVA and MHC class I (H2‐k1) gene expression, consistent with decreased antigen presentation; tumour re‐emergence was also accompanied by decreased PD‐L1 (Cd274) gene expression  cells that contribute to immunosurveillance and the suppression of solid tumour formation  subset  sarcoma cells, but importantly, not HER‐2‐negative 24JK control cells  and central memory (CD44hiCD62Lhi) PTPN2‐deficient CD8+ HER‐2 CAR T cells were more effective at specifically killing 24JK‐HER‐2 cells but not 24JK cells in vitro  of purified Ptpn2fl/fl versus Lck‐Cre;Ptpn2fl/fl central memory CD8+ HER‐2 CAR T cells into sub‐lethally irradiated syngeneic recipients bearing established orthotopic tumours arising from the injection of HER‐2‐expressing E0771 (HER‐2‐E0771) mammary tumour cells  and IL‐10 (Il10)  of the maximal ethically permissible mammary tumour burden prior to CAR T‐cell therapy  rather than the mixed central and effector/memory (CD44hiCD62Llo) phenotypes otherwise present on day 10 post‐adoptive transfer . When CD8+ T cells are activated by a strong TCR stimulus and subsequently stimulated with IL‐2, they undergo differentiation into effectors and acquire CTL activity characterised by IFNγ and granzyme B expression . Since IL‐2‐induced STAT‐5 signalling was only partially corrected in Lck‐Cre;Ptpn2fl/fl;Lck+/− CAR T cells  being reduced to those in Ptpn2fl/fl control CAR T cells    Fig B. To asset al, et al, Ptpn2 in murine CAR T cells by RNA interference using nuclease resistant siRNA duplexes (siSTABLE™) that efficiently knockdown genes for prolonged periods   Fig F. By conet al, ex vivo so that resultant CAR T cells do not overexpress Cas9 and do not elicit immunogenic responses post‐adoptive transfer. To this end, we transfected total CAR T cells with recombinant nuclear‐localised Cas9 pre‐complexed with short guide (sg) RNAs capable of directing Cas9 to the Ptpn2 locus  and the human melanoma cell line MDA‐MB‐435 (ATCC®HTB‐219™) were obtained from the ATCC, Manassas, Virginia, USA. HER‐2‐E0771 cells were engineered to inducibly overexpress murine PTPN2 (HER‐2‐E0771‐PTPN2hi) in response to doxycycline using the Tet‐On 3G Inducible Expression System according to the manufacturer's instructions (Clontech). Tumour cells were cultured in RPMI 1640  or high‐glucose DMEM (AT3‐OVA) supplemented with 10% FBS, l‐glutamine (2 mM), penicillin (100 units/ml)/streptomycin (100 μg/ml), MEM non‐essential amino acids (0.1 mM), sodium‐pyruvate (1 mM), HEPES (10 mM) and 2‐mercaptoethanol (50 μM).The C57BL/6 mouse mammary carcinoma cell line E0771  (Johnstone PtprcaPepcb/BoyJ) and human HER‐2 transgenic (TG) recipient mice and 6‐ to 8‐week‐old female donor mice were used for adoptive transfers. For ex vivo experiments, either male or female mice were used. Aged‐ and sex‐matched littermates were used in all experiments. Ptpn2fl/fl and Lck‐Cre;Ptpn2fl/fl mice and the corresponding OT‐1 TCR transgenic mice were described previously  mice have been described previously  with Lck‐Cre;Ptpn2fl/fl mice. B6.129S2‐Lck<tm1Mak>/J mice were a gift from Dr Andre Veillette  and were bred with Lck‐Cre;Ptpn2fl/fl mice to generate Lck‐Cre;Ptpn2fl/fl;Lck+/− mice. Ly5.1 and C57BL/6 mice were purchased from the WEHI Animal Facility , and human HER‐2 (C57BL/6) transgenic (TG) mice were bred and maintained at the Peter MacCallum Cancer Centre.Mice were maintained on a 12‐h light–dark cycle in a temperature‐controlled high barrier facility with free access to food and water. Six‐ to 10‐week‐old female Ly5.1  was purchased from Thermo Fisher Scientific. The mouse antibody against PTPN2 (clone 6F3) was provided by M. Tremblay (McGill University). For immunofluorescence staining and immunohistochemistry, rabbit anti‐CD3ε and mouse anti‐HER‐2 from Abcam were used. Recombinant human IL‐2, murine IL‐7 and IL‐15 used for T‐cell stimulation or IFNγ used for stimulating tumour cells were purchased from the NIH or PeproTech, respectively. RetroNectin was purchased from Takara, Dnase I and doxycycline from Sigma‐Aldrich. The mouse anti‐nuclear antibodies Ig's  ELISA Kit (Alpha Diagnostic Int.) and the Transaminase II Kit  were used according to the manufacturer's instructions. FBS was purchased from Thermo Scientific; Dulbecco‐Phosphate‐Buffered Saline (D‐PBS), RPMI 1640, DMEM, MEM non‐essential amino acids and sodium‐pyruvate were from Invitrogen, and collagenase type IV was purchased from Worthington Biochemical.For cell stimulation, anti‐CD3ε (clone 145‐2C11) and anti‐CD28 (clone 37.51) antibodies were purchased from BD Biosciences. Antibodies against p‐(Y701) STAT1 (clone 58D6), p‐(Y694) STAT5 (D47E7) XPet al, et al, Single‐cell suspensions from spleen, lymph nodes and hepatic lymphocytes were obtained as previously described  or CyAn™ ADP (Beckman Coulter).For FACS sorting, cells were stained in 15‐ml Falcon tubes (BD Biosciences) for 30 min on ice and purified using either a BD Influx cell sorter, or the BD FACSAria II, BD FACSAria Fusion 3 or BD FACSAria Fusion 5 instruments.Data were analysed using FlowJo8.7 or FlowJo10 (Tree Star Inc.) software. For cell quantification, a known number of Calibrite™ Beads (BD Biosciences) or Nile Red Beads (ProsiTech) or Flow‐Count Fluorospheres (Beckman Coulter) were added to samples before analysis.The following antibodies from BD Biosciences, BioLegend or eBioscience were used for flow cytometry: Phycoerythrin (PE) or peridinin–chlorophyll cyanine 5.5 (PerCP‐Cy5.5)‐conjugated CD3 (145‐2C11); PerCP‐Cy5.5 or phycoerythrin–cyanine 7 (PE‐Cy7)‐conjugated CD4 (RM4‐5); BV711, Pacific Blue‐conjugated (PB), allophycocyanin (APC)‐Cy7 or APC‐conjugated CD8 (53‐6.7); PerCP‐Cy5.5, PE or APC‐Cy7‐conjugated CD25 (P61); Fluorescein isothiocyanate (FITC), V450, Alexa Fluor 700 or PE‐Cy7‐conjugated CD44 (IM7); APC‐conjugated CD45 (30‐F11); V450, APC, Alexa Fluor 488 or PE‐conjugated CD45.1 ; FITC, PB or PerCP‐Cy5.5‐conjugated CD45.2 ; APC‐conjugated CD45R ; PE‐Cy7, APC or PE‐conjugated CD62L (Mel‐14); PE‐conjugated CD122 (TM‐β1); PE‐conjugated CD132 (4G3); Alexa Fluor 647‐conjugated CD183 (CXCR3‐173); PerCP‐eFluo710‐conjugated CD185 ; PE‐Cy7 conjugated CD197 ; PE‐conjugated CD223 ; PE‐Cy7‐conjugated CD279 ; FITC‐conjugated CD11b (M1/70); APC‐Cy7‐conjugated Ly6C (AL‐21); PE‐conjugated Ly‐6G (clone 1A8); APC‐conjugated F4/80 (BM8); PE‐conjugated IL‐10 (JES5‐16E3), PE‐conjugated Nur77 (12.14); PE‐Cy7‐conjugated IFNγ (XMG1.2); FITC or APC‐conjugated TNF (MP6‐XT22); Alexa Fluor 647‐conjugated Granzyme B (GB11); PE‐Cy7‐conjugated TCR‐Vα2 (B20.1); eFluor 660‐conjugated p(Y418)‐Src (SC1T2M3); Alexa Fluor 647‐conjugated T‐bet (4B10); and V450‐conjugated FoxP3 (clone MF23).The following antibodies from Miltenyi Biotec were used for flow cytometry: PE‐conjugated CD197 ; VioBright FITC‐conjugated CD8 (BW135/80); and VioBlue‐conjugated CD45RA (REA1047).7/ml) were cultured overnight with anti‐CD3ε (0.5 μg/ml) and anti‐CD28 (0.5 μg/ml) in the presence of 100 IU/ml human IL‐2 and 0.2 ng/ml murine IL‐7 in complete T‐cell medium . Dead cells were removed by Ficoll centrifugation  according to the manufacturer's instructions. Retrovirus encoding a second‐generation chimeric antibody receptor (CAR) consisting of an extracellular scFv‐anti‐human HER‐2, a membrane proximal CD8 hinge region and the transmembrane and the cytoplasmic signalling domains of CD28 fused to the cytoplasmic region of CD3ζ (scFv‐anti‐HER‐2‐CD28‐ζ) was obtained from the supernatant of the GP+E86 packaging line as described previously  at room temperature and incubated overnight before the second viral transduction. T cells were maintained in IL‐2‐ and IL‐7‐containing media and cells used at days 7–8 after transduction.Splenocytes were isolated from murine spleens by mechanical disruption, and contaminating red blood cells were removed by incubation with 1 ml of Blood Cell Lysing Buffer Hybri‐Max™ (Sigma‐Aldrich) for 7 min at room temperature per spleen. Lymphocytes  CAR T cells and expanded for up to 15 days in the presence of human IL‐2. The expression of the chimeric receptor was determined by staining with Alexa Flour 647‐conjugated anti‐3S193 idiotype [supplied by Ludwig Institute for Cancer Research (LICR)], and truncated CD34 was detected with PE‐conjugated anti‐human CD34 (BioLegend). Cell surface phenotyping of transduced cells was determined by staining with BV785‐conjugated anti‐human CD3 , BV605‐conjugated anti‐human CD4  and PE‐Cy7‐conjugated anti‐human CD8 .Human peripheral blood mononuclear cells (PBMCs) were isolated from normal donor buffy coats. PBMCs were stimulated with anti‐human CD3  and human IL‐2 (600 IU/ml) in complete T‐cell medium. Retrovirus encoding a second‐generation chimeric antibody receptor (CAR) consisting of an extracellular scFv‐anti‐human LeY domain, a membrane proximal CD8 hinge region and the transmembrane and the cytoplasmic signalling domains of CD28 fused to the cytoplasmic region of CD3ζ was obtained from the supernatant of the PG13 packaging cell line and has been described previously (Westwood 5) generated from human PBMC were incubated with Lewis Y‐expressing OVCAR‐3 cells (1 × 105) or Lewis Y‐negative MDA‐MB‐435 cells (1 × 105) or plate‐bound anti‐LY  for 6 h at 37°C in complete T‐cell medium. GolgiPlug™ and GolgiStop™ were added 3 h before cells were processed for surface and intracellular staining for CD4 , CD8 , IFNγ  and TNF  by flow cytometry.CAR T cells  or IL‐15 (5 or 10 ng/ml) in 100 μl RPMI/1 %FBS. Cells were processed for intracellular p(Y694)‐STAT‐5 detection by flow cytometry as previously described  dilution, purified CD8+CD44hiCD62Lhi CAR T cells were incubated with CTV in D‐PBS supplemented with 0.1% (v/v) BSA at a final concentration of 2 μM for 10 min at 37°C. Cells were then washed three times with D‐PBS supplemented with 10% (v/v) FBS. CAR T cells (2 × 105) were incubated with HER‐2‐expressing or HER‐2‐negative 24JK sarcoma cells (1 × 105) in complete T‐cell medium.CAR T cells generated from mouse splenocytes were stained with fluorochrome‐conjugated antibodies for CD8, CD62L and CD44 and sorted for CD8+CD44hiCD62Lhi CAR T cells were incubated with plate‐bound anti‐CD3ε for 24 h at 37°C in complete T‐cell medium. Pre‐activated CAR T Cells (2 × 105) were labelled with 5 μM CTV and incubated with HER‐2‐expressing or HER‐2‐negative 24JK sarcoma cells (1 × 105) in complete T‐cell medium. At various time points, proliferating cells were harvested and CTV dilution was monitored by flow cytometry. For cell quantification, Nile Red Beads (ProsiTech) or Flow‐Count Fluorospheres (Beckman Coulter) were added to samples before analysis.For the assessment of CAR T‐cell proliferation after anti‐CD3ε stimulation, purified CD8T cells were incubated rocking on an orbital shaker with PTPN2‐inhibitor compound 8 (100 nM) or vehicle (DMSO 1% v/v) in serum‐free RPMI 1640 for 1 h at 37°C. Cells were washed three times with complete T‐cell medium and processed for T‐cell activation studies.5 HER‐2‐E0771 cells resuspended in 20 μl D‐PBS into the fourth mammary fat pad. At day 6 post‐tumour cell injection, human HER‐2 TG mice were pre‐conditioned with total body irradiation (4 Gy) prior to the adoptive transfer of 6 × 106 FACS‐purified CD8+CD44hiCD62Lhi CAR T cells. Mice were treated with 50,000 IU IL‐2 on days 0–4 after T‐cell transfer.Female human HER‐2 TG mice were anaesthetised with Ketamine (100 mg/kg) and Xylazil (10 mg/kg) (Troy Laboratories) and injected orthotopically with 2 × 102) was abraded using a MultiPro Dremel with a grindstone attachment. B16.F10‐OVA‐expressing melanoma cells (1 × 105) were resuspended in 10 μl of Matrigel™ (BD Biosciences) and orthotopically applied to the lesion. Set Matrigel was covered with a piece of Op‐site FlexigridTM (Smith and Nephew). The torsos of the mice were wrapped with a soft hypoallergenic MicroporeTM tape  and then by a stronger porous polyethylene TransporeTM tape  to protect the abraded site. Twenty‐four hours later, 2 × 105 FACS‐purified CD8+ naive CD44loCD62Lhi OT‐1 T cells were adoptively transferred. Recipient mice were monitored daily until bandages were removed 6 days after tumour engraftment.Ly5.1 mice were anaesthetised with Ketamine (100 mg/kg) and Xylazil (10 mg/kg) (Troy Laboratories). The flanks of the mice were shaved using clippers and depilated with Veet (Reckitt Benckiser), and the skin  and Xylazil (10 mg/kg) (Troy Laboratories) and were injected orthotopically with 1 × 106/200 μl) were stimulated with PMA  and ionomycin  in the presence of GolgiPlug™ and GolgiStop™ (BD Biosciences) for 4 h at 37°C in complete T‐cell medium.Tumour‐bearing mice were sacrificed, and tumours were excised and digested at 37°C for 30 min using a cocktail of 1 mg/ml collagenase type IV  and 0.02 mg/ml DNase (Sigma‐Aldrich) in DMEM supplemented with 2% (v/v) FBS. Cells were passed through a 70‐μm cell strainer (BD Biosciences) twice and processed for flow cytometry. For the detection of intracellular cytokines in tumour‐infiltrating lymphocytes, cells  and 10 mM HEPES for 1 h at 37°C. Cells were passed through a 70‐μm cell strainer twice  and washed three times with 50 ml PBS. Cells were resuspended in DMEM supplemented with 10% FBS, 2 mM l‐glutamine, penicillin (100 units/ml), streptomycin (100 μg/ml) and seeded into 24‐well plates. Cells were incubated at 37°C until they reached 80% confluency. Tumour‐infiltrating T cells from AT‐3‐OVA tumour‐bearing C57BL/6 mice Ptpn2fl/fl and Lck‐Cre;Ptpn2fl/fl mice were labelled with 2 μM CTV and incubated with adherent, 80% confluent AT‐3‐OVA tumour cells overnight. GolgiPlug™ and GolgiStop™ were added 3 h before cells were processed for intracellular staining for IFNγ and TNF by flow cytometry.AT‐3‐OVA tumour‐bearing C57BL/6 mice were sacrificed, and tumours were collected in ice‐cold D‐PBS and then digested with 50 μg/ml Liberase  in Hank's Balanced Salt Solution  or effector/memory (CD44hiCD62Llo) or total HER‐2‐specific CAR T cells were added at different concentrations to the mix of HER‐2‐expressing (5 × 104) and HER‐2‐negative (5 × 104) 24JK sarcoma cells and incubated for 4 h at 37°C in complete T‐cell medium. Antigen‐specific target cell lysis (24JK‐HER‐2 cell depletion) was monitored by flow cytometry.For the assessment of CAR T‐cell cytotoxicity, HER‐2‐expressing 24JK sarcoma cells (5 μM CTV) and HER‐2‐negative 24JK sarcoma cells (0.5 μM CTV) were labelled with CellTrace™ Violet  in D‐PBS supplemented with 0.1% (v/v) BSA for 15 min at 37°C. Tumour cells were then washed three times with D‐PBS supplemented with 10% (v/v) FBS and mixed at a 1:1 ratio. FACS‐purified CD8For brain immunohistochemistry, mice were perfused transcardially with heparinised saline  followed by 4% (w/v) paraformaldehyde in phosphate buffer . Brains were post‐fixed overnight and then kept for 4 days in 30% (w/v) sucrose in 0.1 M phosphate buffer to cryoprotect the tissue, before freezing on dry ice. 30‐μm sections (120 μm apart) were cut in the coronal plane throughout the entire rostral‐caudal extent of the cerebellum. For detection of CD3ε, sections were subjected to antigen retrieval in citrate acid buffer  at 85°C for 20 min. Sections were incubated at room temperature for 2 h in blocking buffer [0.1M phosphate buffer, 0.2% (v/v) Triton X‐100, 10% (v/v) normal goat serum (Sigma‐Aldrich) and then overnight at 4°C in rabbit anti‐CD3 (Dako) in 1% (v/v) blocking buffer. After washing with PBS, sections were incubated with goat anti‐rabbit Alexa Fluor 488‐conjugated secondary antibody (Life Technologies) in blocking buffer for 2 h at room temperature. Sections were mounted with Mowiol 4‐88 mounting media and visualised using an Olympus Provis AX70 microscope. Images were captured with an Olympus DP70 digital camera and processed using AnalySIS (Olympus) software. To assess gross tissue morphology, a subset of sections throughout the entire rostral‐caudal extent of the cerebellum were stained with haematoxylin and eosin.For detection of HER‐2, 30‐μm sections throughout the entire rostral‐caudal extent of the cerebellum from C57BL/6 or HER‐2 TG C57BL/6 mice were subjected to antigen retrieval and incubated in blocking buffer (as described above). Sections were then incubated overnight in (4°C) with anti‐c‐ErbB2/c‐Neu (Ab‐3) mouse mAb (3B5) . After washing with PBS, HER‐2‐positive cells were visualised using rabbit IgG VECTORSTAIN ABC Elite and DAB  Peroxidase Substrate Kits  and visualised using a bright field microscope.For mammary fat pad tumour immunohistochemistry, animals were culled and mammary fat pad immediately dissected and fixed in buffered formalin solution for 48 h. Tissues were embedded in paraffin and 4‐μm sections of the entire block prepared. Every tenth to fourteenth section of the tissue was used to detect HER‐2 and CD3 by immunohistochemistry. After deparaffinisation and rehydration, sections were subjected to antigen retrieval in Tris/EDTA buffer (pH 8.0) at 120°C for 10 min. Sections were blocked with 5% (v/v) bovine serum albumin in 0.1 M phosphate buffer for 1 h at room temperature and incubated overnight (4°C) with anti‐c‐ErbB2/c‐Neu (Ab‐3) mouse mAb (3B5)  or anti‐CD3 . After washing with PBS, HER‐2‐ and CD3‐positive cells were visualised using rabbit IgG VECTORSTAIN ABC Elite and DAB  Peroxidase Substrate Kits  and counterstained with haematoxylin. Sections were visualised on a Zeiss Axioskop 2 mot plus microscope .™ SYBR® Green Assay  were utilised to perform quantitative PCR detecting Cxcl9, Cxcl10, Tgfb1, Cd274, H2‐K1 and Rps18. Primer sets for HER‐2, Nono and SerpinB14  were purchased from Sigma‐Aldrich. Relative gene expression (ΔCt) was determined by normalisation to the house‐keeping gene Nono throughout the study except for Fig Rps18 was used and ΔΔCt analysis performed.RNA was extracted with TRIzol reagent  and RNA quality and quantity determined using a NanoDrop 2000 (Thermo Fisher Scientific). mRNA was reverse transcribed using a High‐Capacity cDNA Reverse Transcription Kit  and processed for quantitative real‐time PCR either using the Fast SYBR™ Green Master Mix . Primer sets from PrimePCRPtpn2 was knocked down transiently in HER‐2 CAR T cells using nuclease resistant Ptpn2 siRNA duplexes  ; siRNA duplexes (siSTABLE™) for enhanced green fluorescent protein  were used as a control. CAR T cells were transfected with Ptpn2 siRNA (300 nM) conjugated to FITC or GFP siRNA (300 nM) conjugated to FITC 2 days prior to adoptive T‐cell therapy using the Mouse T‐cell Nucleofector™ Kit (Lonza Bioscience) according to the manufacturer's instructions.Ptpn2 was deleted in HER‐2 CAR T cells using Cas9 ribonucleoprotein (RNP)‐mediated gene editing. Briefly, total CAR T cells were transfected with recombinant Cas9  pre‐complexed with short guide (sg) RNAs  targeting the Ptpn2 locus  or non‐targeting sgRNAs (GCACUACCAG AGCUAA CUCA) as a control 2 days prior to adoptive T‐cell therapy using the P3 Primary Cell 4D‐Nucleofector X™ Kit (Lonza Bioscience) according to the manufacturer's instructions.hi cell line. Briefly, murine Ptpn2 cDNA from HER‐2‐E0771 cells was reverse transcribed and amplified by PCR and cloned into the SmaI and EcoR1 restriction sites of Tre‐3G plasmid (Clontech Laboratories) to generate the Tre‐3G‐Ptpn2 construct; the fidelity of the cloned cDNA was confirmed by dideoxy sequencing. HEK293T cells were transfected with either the Tre‐3G‐Ptpn2 or Tet‐On 3G constructs using the Lenti‐X Packing system (Clontech Laboratories) according to the manufacturer's instructions. HER‐2‐E0771 mammary tumour cells were transduced first with Tet‐On 3G lentivirus and selected with G418 (0.8 mg/ml) and subsequently transduced with Tre‐3G‐Ptpn2 lentivirus and selected with puromycin (1.5 μg/ml). Where indicated, the resultant HER‐2‐E0771‐PTPN2hi cells were incubated with 2 mg/ml doxycycline (DOX) to induce the expression of PTPN2. Cells were serum starved in DMEM medium without FBS for 12 h and stimulated with 2 ng/ml IFNγ for the indicated times and processed for immunoblotting or incubated with 1 ng/ml IFNγ for 24 h and processed for quantitative real‐time PCR.The Tet‐on 3G inducible expression system (Clontech Laboratories) was used to generate the HER‐2‐E0771‐PTPN2in vivo studies, 2 × 105 HER‐2‐E0771‐PTPN2hi cells were resuspended in 20 μl D‐PBS and injected orthotopically into the fourth mammary fat pad of female human HER‐2 TG mice. At day 5 post‐tumour cell injection, mice were administered DOX (2 mg/ml) in the drinking water for the entirety of the experiment. At day 6 post‐tumour cell injection, human HER‐2 TG mice were pre‐conditioned with total body irradiation (4 Gy) prior to the adoptive transfer of 6 × 106 FACS‐purified CD8+CD44hiCD62Lhi CAR T cells. Mice were treated with 50,000 IU IL‐2 on days 0–4 after T‐cell transfer.For U‐test, the parametric 2‐tailed Student's t‐test, the 1‐way or 2‐way ANOVA test using Turkey or Sidak post hoc comparison or the log‐rank (Mantel–Cox test) where indicated. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001 were considered as significant.Statistical analyses were performed with GraphPad Prism software 7.0b using the non‐parametric using 2‐tailed Mann–Whitney All experiments were performed in accordance with the NHMRC Australian Code of Practice for the Care and Use of Animals. All protocols were approved by the Monash University School of Biomedical Sciences Animal Ethics Committee (Ethics number: MARP/2012/124) or the Peter MacCallum Animal Ethics and Experimentation Committee .Conceptualisation: TT; Methodology: FW, K‐HL, XD, SL, KH, GTD, PKG, CK, DM, PAB, MAH, SLP, JW, SZ, Z‐YZ, JO, TG, PKD and TT; Data acquisition: FW, K‐HL, XD, SL, KH, GTD, PKG, MAH and SLP; Data analysis: FW, K‐HL, XD, SL, KH, GTD, PKG; MAH; TG, PKD and TT; Data interpretation: FW, K‐HL, XD, SL, KH, GTD, PKG, CK, DM, PAB, MAH, SLP, JW, SZ, Z‐YZ, JO, TG, PKD and TT; Writing‐Original Draft: TT; Writing‐Review and Editing: FW, K‐HL, XD, SL, KH, GTD, PKG, CK, DM, PAB, MAH, SLP, JW, SZ, Z‐YZ, JO, TG, PKD and TT. Funding acquisition: FW, PKD, TG and TT; Final approval for publishing: FW, K‐HL, XD, SL, KH, GTD, PKG, CK, DM, PAB, MAH, SLP, JW, SZ, Z‐YZ, JO, TG, PKD and TT.The authors declare that they have no conflict of interest.AppendixClick here for additional data file.Expanded View Figures PDFClick here for additional data file.Source Data for AppendixClick here for additional data file.Review Process FileClick here for additional data file.


The title compound, 2-(4-nitrophenyl)-2-oxoethyl 2-chlorobenzoate, is relatively planar with the two aromatic rings being inclined to each other by 3.56 (11)°. 15H10ClNO5, is relatively planar with the two aromatic rings being inclined to each other by 3.56 (11)°. The central —C(=O)—C–O—C(=O)— bridge is slightly twisted, with a C—C—O—C torsion angle of 164.95 (16)°. In the crystal, molecules are linked by C—H⋯O and C—H⋯Cl hydrogen bonds, forming layers parallel to the (101) plane. The layers are linked by a further C—H⋯O hydrogen bond, forming a three-dimensional supramolecular structure. There are a number of offset π–π interactions present between the layers [intercentroid distances vary from 3.8264 (15) to 3.9775 (14) Å]. Hirshfeld surface analyses, the dnorm surfaces, electrostatic potential and two-dimensional fingerprint plots were examined to verify the contributions of the different intermolecular contacts within the supramolecular structure. The shape-index surface shows that two sides of the molecule are involved in the same contacts with neighbouring molecules, and the curvedness plot shows flat surface patches that are characteristic of planar stacking.The title compound, C Torsion angle τ1 at −12.5 (3)° signifies a certain noncoplanarity between the benzene ring (C10–C15) and the adjacent carbonyl group (C9=O3) as a result of steric repulsion between the substituent Cl1 and the adjacent carbonyl group C9=O3. This is also reflected in the torsion angle τ2 of −164.95 (16)°, between the two carbonyl groups, C7=O2 and C9=O3, which have a –antiperiplanar conformation. Torsion angle τ3, involving the benzene ring (C1–C6) and the adjacent carbonyl group (C7=O2), is −3.6 (3)° and indicates a –synperiplanar conformation. A comparison of the torsion angles in I and II, indicates that the insertion of the Cl atom in I has the most significant influence on torsion τ2, which is −164.95 (16)° in I compared to 174.08 (9)° in II. Torsion angles τ1 of −12.5 (3)° and τ3 of −3.6 (3)° are slightly larger than the values observed in II, viz. 9.60 (16) and 1.88 (15)°, respectively. Hence, compound I has a less planar conformation than unsubstituted compound II.The overall molecular conformation of I is characterized by three torsion angles, i, C14—H14⋯O4i and C13—H13⋯Cl1iii hydrogen bonds to form layers lying parallel to the (101) plane; see Fig. 2A⋯O3ii hydrogen bonds and offset π–π interactions  shows the Hirshfeld surface mapped over dnorm (−0.154 to 1.305) and for Fig. 4b) the electrostatic potential. The Hirshfeld surface illustrated in Fig. 4a) reflects the involvement of different atoms with the intermolecular interactions through the appearance of blue and red patches, which correspond to the regions of positive and negative electrostatic potential shown in Fig. 4b). The shape-index surface  clearly shows that the two sides of the molecule are involved in contacts with neighbouring molecules and the curvedness plot  shows flat surface patches characteristic of planar stacking.The Hirshfeld surface analysis -2-oxoethyl 4-nitrobenzoate ° in the title compound. In the crystal structure of the recently published compound 2-(4-nitrophenyl)-2-oxoethyl benzoate (II)  = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989019014336/su5521sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989019014336/su5521Isup2.hklStructure factors: contains datablock(s) I. DOI: 1449647, 1449647CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In the crystal, phenyl-C—H⋯O(carbonyl) and carbonyl-O⋯O(carbonyl) interactions general a [111] supramolecular chain.In Ru 3(C19H17PS)(CO)11], comprises a triangle of Ru0 atoms, two of which are bonded to four carbonyl ligands. The third metal atom is bound to three carbonyl ligands and the phosphane-P atom of a dissymmetric phosphane ligand, PPh2(C6H4SMe-4); no Ru⋯S interactions are observed. The phosphane occupies an equatorial position and its proximity to an Ru—Ru edge results in the elongation of this bond with respect to the others [2.8933 (2) Å cf. 2.8575 (2) and 2.8594 (3) Å]. In the crystal, phenyl-C—H⋯O(carbonyl) and carbonyl-O⋯O(carbonyl) [2.817 (2) Å] interactions combine to form a supramolecular chain propagating along [111]; the chains pack without directional interactions between them. The carbonyl-O⋯O(carbonyl) and other weak contacts have an influence upon the Hirshfeld surfaces with O⋯H contacts making the greatest contribution, i.e. 37.4% cf. 15.8% for O⋯O and 15.6% for H⋯H contacts.The title cluster compound, [Ru R3) have played a major role in the formation and subsequent chemistry of metal carbonyl clusters, often relating to the promising catalytic activity of the products 12 with PR3 leads to Ru3(CO)n12 – (PR3)n, n = 1–4, cluster compounds 12, thereby making the cluser more reactive 10[PPh2(C6H4SMe)]8 cluster, only the P atom of the PPh2C6H4SMe ligand is coordinated to the metal centre while the thiomethyl group remains uncoordinated  ligand are surprisingly few in number 11PPh2(C6H4SMe-4) (I)Tertiary phosphanes (P3(CO)11PPh2(C6H4SMe-4), (I)3 triangle with one Ru centre being bound, equatorially, by the phosphane ligand. The Ru—Ru bond lengths in the Ru3 triangle are not equivalent with the Ru1—Ru2 bond of 2.8933 (2) Å being longer than the Ru1—Ru3 and Ru2—Ru3 bonds of 2.8575 (2) and 2.8594 (3) Å, respectively. This disparity probably reflects the steric hindrance exerted by the phosphane ligand which occupies the region in the vicinity of the Ru1—Ru2 bond. Some general trends in the geometric parameters involving the carbonyl ligands may be discerned, the relatively high errors in some of the parameters notwithstanding. Thus, the Ru—C bond distances involving carbonyl groups lying in the plane of the Ru3 ring are generally shorter than those occupying positions perpendicular to the plane, with the respective ranges in Ru—C bond lengths being 1.897 (3)–1.930 (3) Å and 1.937 (2)–1.953 (3) Å. While the Ru—C≡O angles are all close to linear, two distinctive ranges in angles are evident. The Ru—C≡O angles involving carbonyl groups lying in the plane of the Ru3 ring lie in the range 177.3 (2)–178.7 (2)° while the range for the perpendicularly orientated carbonyl groups is 172.1 (2)–174.6 (2)°. The trend for longer Ru—C distances and greater deviations from linearity of the Ru—C≡O angles for the axial carbonyl ligands, which occupy positions trans to other carbonyl ligands, is consistent with some semi-bridging character for these carbonyl ligands. Thus, the closest intramolecular Ru⋯C(carbonyl) contact of 3.233 (3) Å is formed by the C8-carbonyl ligand which exhibits the maximum deviation from linearity, i.e. 172.1 (2)°.The molecular structure of Rua. The O3⋯O3i separation is 2.817 (2) Å, a distance less than the sum of the van der Waals radii of oxygen, i.e. 3.04 Å : 1 − x, 1 − y, 1 − z. Such intermolecular O⋯O interactions are examples of homoatomic chalcogen bonding which are rarest for the smaller oxygen atoms et al., 2017dnorm are shown in Fig. 3a, results from the C21—H⋯O8 interaction  contact is evident. Carbonyl⋯π(arene) interactions are known to be important in the structural chemistry of metal carbonyls  separation is 3.850 (3) Å and the angle subtended at the O4 atom is 90.1 (2)°, indicating a side-on  approach between the residues. The environment about a reference molecule, showing short interatomic O⋯O and C⋯H/H⋯C contacts significant in the molecule packing of (I)The Hirshfeld surface calculations of (I)n Table 1. The preet al., 2007i.e. 15.6%, contribution from these contacts to the Hirshfeld surfaces is due to the presence of the carbonyl groups on the Ru-cluster which leads to an increase in the contribution of O⋯H/H⋯O contacts to the Hirshfeld surface, i.e. 37.4%. The single tip at de + di ∼2.4 Å in the H⋯H delineated fingerprint plot, which has a broad appearance, arises from a van der Waals contact between the methyl-H18B and phenyl-H20 atoms s Table 2. The twoe Table 1 and a she Table 1, respecte Table 1 and othee Table 1 are views Table 2 result iChemical context, there are two other Ru3 clusters in the literature having the same (4-methylsulfanylphenyl)diphenylphosphane ligand as in (I)3(CO)9PPh2(C6H4SMe-4)(Ph2PCH2PPh2) (II) 9PPh2(C6H4SMe-4)(Ph2AsCH2AsPh2) 11PRR′R′′ and several examples where the phosphane ligand is bidentate bridging, i.e. Ru3(CO)11PR(R′)–R′′–(R′)RPRu3(CO)11 11PRR′R′′.As mentioned in the 3(CO)12 was purchased from Aldrich and PPh2C6H4SMe was synthesized as reported previously 11P(C6H4SMe-4)Ph2 (I)3(CO)12  and PPh2(C6H4SMe)  in tetrahydrofuran (25 ml). The reaction mixture was treated dropwise with sodium diphenylketyl solution until the colour of the mixture turned from orange to dark red and then stirred for 30 min. The solvent was evaporated under vacuum and the residue was chromatographed by preparative TLC. Elution with 7:3 n-hexane/dichloromethane mixture gave four bands and the major orange fraction was characterized as (I)30H17O11PRu3S: C, 39.18; H, 1.86%. Found: C, 39.60; H, 1.90%. IR (C6H12): ν(CO) 2097(m), 2059(w), 2046(m), 2015(s), 1989(w) cm−1. 1H NMR (CDCl3): δ 7.45–7.23 , 2.48 . 13C NMR (CDCl3): δ 204.24 (Ru—CO), 135.19–125.37 (Ph), 14.79 (Me). 31P NMR (CDCl3): δ 34.28 (s).All reactions were carried out under an inert atmosphere of oxygen-free nitrogen (OFN) using standard Schlenk techniques. RuUiso(H) set to 1.2–1.5Ueq(C). Owing to poor agreement, four reflections, i.e. (1 7 14),  I, global. DOI: 10.1107/S2056989018006989/hb7749Isup2.hklStructure factors: contains datablock(s) I. DOI: 1842044CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The title compound is constructed from two aromatic rings , which are linked by a C=C—C(=O)—C enone bridge and form a dihedral angle of 17.91 (17)°. In the crystal, molecules are linked by C—H⋯O hydrogen bonds enclosing rings of  17H14BrFO3, the aromatic rings are tilted with respect to the enone bridge by 13.63 (14) and 4.27 (15)°, and form a dihedral angle 17.91 (17)°. In the crystal, centrosymmetrically related molecules are linked by pairs of C—H⋯O hydrogen bonds into dimeric units, forming rings of R22(14) graph-set motif. The dimers are further connected by weak C—H⋯O hydrogen interactions, forming layers parallel to (10In the molecule of the title compound, C Chalcones, which are considered to be the precursors of flavonoids and isoflavonoids, are abundant in edible plants. They consist of open-chain flavonoids in which the two aromatic rings are joined by a three-carbon α,β-unsaturated carbonyl system. These are coloured compounds because of the presence of the –CO—CH=CH– chromophore, the colour depending on the presence of other auxochromes. Accumulating evidence has shown that chalcones and their derivatives could inhibit tumor initiation and progression. In view of the above and in a continuation of our previous work on 3,4-dimethoxy chalcones  Å for C16 and −0.124 (4) Å for C17. The bond lengths and angles are comparable with those found in the related compounds (2E)-3-(3-chlorophenyl)-1--prop-2-en-1-one -3--1-prop-2-en-1-one -1-(3-bromophenyl)-3-prop-2-en-1-one -3-(2-bromophenyl)-1-prop-2-en-1-one  Å, C14⋯Cg1i = 4.010 (4) Å, C14—Br1⋯Cg1i = 82.54 (11)°; symmetry code: (i) −1 + x, y, z; Cg1 is the centroid of the C1–C6 ring] and C—F⋯π  interactions help to stabilize the crystal structure.In addition, weak C—Br⋯π [C14—Br1 = 1.877 (3) Å, Br1⋯CrystalExplorer17.5 . The O⋯H/H⋯O, Br⋯C/C⋯Br and F⋯C/C⋯F contacts in the structure with 17.9, 5.6 and 5.0% contributions, respectively, to the Hirshfeld surface have a symmetrical distribution of points . The other Br⋯C / C⋯Br, F⋯C / C⋯F, C⋯C, F⋯O / O⋯F and C⋯O / O⋯C contacts, having only small contributions to the Hirshfeld surface, have negligible directional impact on the molecular packing.Molecular Hirshfeld surfaces  = 1.2Ueq(C) or 1.5Ueq(C) for methyl H atoms.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989018009416/rz5240sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989018009416/rz5240Isup2.hklStructure factors: contains datablock(s) I. DOI: 1852842CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


In the crystal, molecules are linked via pairs of the weak intermolecular C—H⋯N hydrogen bonds, forming inversion dimers with b]pyridine ring systems.The imidazopyridine unit linked to phenyl and allyl substituents. The allyl substituent is rotated significantly out of the imidazopyridine plane, while the benzene ring is inclined by 3.84 (6)° to the ring system. In the crystal, molecules are linked via a pair of weak intermolecular C—H⋯N hydrogen bonds, forming an inversion dimer with an R22(20) ring motif. The dimers are further connected by π–π stacking interactions between the imidazopyridine ring systems [centroid–centroid distances = 3.7161 (13) and 3.8478 (13) Å]. The important contributions to the Hirshfeld surface are H⋯H (35.9%), H⋯Cl/Cl⋯H (15.0%), H⋯C/C⋯H (12.4%), H⋯Br/Br⋯H (10.8%), H⋯N/N⋯H (7.5%), C⋯Br/Br⋯C (5.9%), C⋯C (5.5%) and C⋯N/N⋯C (4.0%) contacts.The title compound, C Their activities include anticancer -4H-imidazopyridine in the presence of a catalytic quantity of tetra-n-butylammonium bromide under mild conditions.As a continuation of our research work devoted to the development of substituted imidazopyridine unit linked to phenyl and allyl substituents  Å for atom C12. The ring system is inclined by 3.84 (6)° to the benzene C1–C6 ring, with the N2—C7—C6—C1 torsion angle being 3.3 (3)°. The allyl substituent is nearly perpendicular to the imidazopyridine plane, as indicated by the C8—N3—C13—C14 torsion angle of −97.3 (2)°. Atoms C6 and C13 are 0.038 (2) and 0.014 (2) Å, respectively, away from the imidazopyridine plane.The title compound is built up from an imidazopyridine ring systems. The centroid–centroid distances, Cg1⋯Cg1ii and Cg1⋯Cg2i , are 3.7161 (13) and 3.8478 (13) Å, respectively, where Cg1 and Cg2 are the centroids of the N1/N2/C7–C9 and N3/C8–C12 rings, respectively.In the crystal, molecules are linked if Fig. 2. The dimpara-substituated analogue, namely 4-allyl-6-bromo-2-phenyl-4H-imidazopyridine monohydrate, has been reported –(l), together with their relative contributions to the Hirshfeld surface. The contributions are 35.9, 15.0, 12.4, 10.8, 7.5, 5.9, 5.5, 4.0, 1.5, 1.2 and 0.2%, respectively, for H⋯H, H⋯Cl/Cl⋯H, H⋯C/C⋯H, H⋯Br/Br⋯H, H⋯N/N⋯H, C⋯Br/Br⋯C, C⋯C, C⋯N/N⋯C, C⋯Cl/Cl⋯C, N⋯Br/Br⋯N and N⋯N contacts. The most important interaction is H⋯H (35.9%), which is reflected as widely scattered points of high density due to the large hydrogen content of the molecule [Fig. 5b)]. The spike with the tip at de = di = 1.16 Å is due to the short interatomic H⋯H contacts. The H⋯Cl/Cl⋯H contacts (15.0%) have a nearly symmetrical distribution of points and a pair of spikes with tips at de + di = 2.67 Å [Fig. 5c)]. In the absence of C—H ⋯ π interactions, the H⋯C/C⋯H contacts (12.4%) also have a nearly symmetrical distribution of points with tips at de + di = 2.79 Å [Fig. 5d)]. The H⋯Br/Br⋯H contacts (10.8%) have a symmetrical distribution of points and a pair of spikes with tips at de + di = 3.00 Å [Fig. 5e)]. A pair of spikes with tips at de + di = 2.42 Å  in the H⋯N/N⋯H contacts (7.5%) arises from the C—H⋯N hydrogen bond ]. The C⋯C contacts (5.5%) have an arrow-shaped distribution of points with the tip at de = di = 1.75 Å [Fig. 5h)]. The C⋯N/N⋯C contacts (4.0%) have wide spikes with tips at de + di = 3.44 Å [Fig. 5i)]. The HS representations with the function dnorm plotted onto the surface are shown for the H⋯H, H⋯Cl/Cl⋯H, H⋯C/C⋯H, H⋯Br/Br⋯H, H⋯N/N⋯H, C⋯Br/Br⋯C, C⋯C and C⋯N/N⋯C contacts [Figs. 6a)–(h)].In order to visualize the intermolecular interactions in the crystal of the title compound, a Hirshfeld surface (HS) analysis  dissolved in 25 ml of N,N-dimethylformamide (DMF) and potassium carbonate  was stirred for 5 min, and then to a mixture of tetra-n-butylammonium bromide  and allyl bromide  was added. Stirring was continued for 6 h at room temperature. After removing the salts by filtration, DMF was evaporated under reduced pressure, and the solid obtained was dissolved in dichloromethane. The residue was extracted with distilled water and the resulting mixture was chromatographed on a silica-gel column . Brown single crystals suitable for X-ray diffraction were obtained by evaporation of an ethyl acetate–hexane (1:3 v/v) solution.A mixture of 6-bromo-2-(4-chlorophenyl)-4Uiso(H) = 1.2Ueq(C).Crystal data, data collection and refinement details are summarized in Table 210.1107/S2056989018017322/is5505sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989018017322/is5505Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989018017322/is5505Isup3.cdxSupporting information file. DOI: Click here for additional data file.10.1107/S2056989018017322/is5505Isup4.cmlSupporting information file. DOI: 1883384CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information: