N‐halogenosuccinimides or N‐halogenophthalimides led to a series of new redox‐active halogeno‐ and succinimido‐/phthalimido‐substituted derivatives. These highly selective reactions are proposed to proceed via the tri‐ or tetracationic state as the intermediate. The derivatives are oxidised reversibly at slightly higher potentials than that of the unsubstituted compounds to dications and further to tri‐ and tetracations. The integration of redox‐active ligands in the transition‐metal complexes shifts the redox potentials to higher values and also allows reversible oxidation in two potentially separated one‐electron steps.New redox‐active 1,2,5,6‐tetrakis(guanidino)‐naphthalene compounds, isolable and storable in the neutral and deep‐green dicationic redox states and oxidisable further in two one‐electron steps to the tetracations, are reported. Protonation switches on blue fluorescence, with the fluorescence intensity (quantum yield) increasing with the degree of protonation. Reactions with  Redox‐active guanidines: New redox‐active guanidines, stable in the neutral as well as dicationic redox states and reversibly oxidisable up to the tetracations, were synthesised. Protonation switches on fluorescence; reactions with N‐halogenosuccinimides (NXS) and ‐phthalimides selectively gives fully substituted derivatives. Coordination of palladium acetate yields binuclear complexes in which the bridging guanidine ligands are oxidisable in two reversible one‐electron steps. There is a huge demand for redox‐active compounds that exhibit two (or more) stable redox states.1, Figure 1 is a relatively strong organic electron donor with a redox potential E1/2 (in CH3CN) versus ferrocenium/ferrocene (Fc+/Fc) of −0.73 V and also a strong Brønsted base with an estimated pKa value of 25.3 for (1+H)+ in CH3CN. A number of stable, storable salts of the oxidised compound, for example, 1(BF4)2, 1(PF6)2, 1(I3)2, 1{N(CN)2}2E1/2) up to approximately −1 V have been since synthesised.I complexes with oxidised GFA ligands and CuII complexes with neutral GFA ligands.In the last decade, we developed guanidino‐functionalised aromatics (GFAs) as a new class of redox‐active organic molecules,2 and 3 Ka values (estimated to be 27.4 for (2+H)+ in CH3CN). For (2+2H)2+, the positions of the protons in (asymmetric) N−H⋅⋅⋅N bridges were confirmed by NMR spectroscopic studies in solution and structural characterisation in the solid state.22+ are known, for example, the salt 2(I3)2 and the complex [2(CuBr2)2] (a dinuclear CuI complex with the dication 22+ as bridging ligand),The two 1,4,5,8‐tetrakis(guanidino)‐naphthalene derivatives 4) and 1,2,5,6‐tetrakis‐naphthalene (5) are reported ‐naphthalene  or 2‐chloro‐1,3‐dimethylimidazolinium chloride to give 5 (52 % isolated yield).The new tetrakisguanidines 4 were grown by diffusing diethyl ether into a saturated acetonitrile solution. Crystals of 5 were obtained from a saturated acetonitrile solution. Figure 4 and 5. The CN3 planes of the guanidino groups are highly twisted  with respect to the aromatic‐ring plane, and the C1‐N1‐C6/C11 and C2‐N4‐C11/C16 bond angles at the nitrogen atoms attached to the aromatic core (120.27(11)° and 119.69(11)° for 4 and 127.20(16)° and 122.95(15)° for 5) display significant deviation from a linear geometry. This “bent‐twisted” structure is in line with the structures of other GFA compounds.3 plane is in the aromatic plane, is disfavoured by steric strain. As a consequence of the “bent‐twisted” conformation, there is no steric strain in molecules with two guanidino groups ortho to each other (in contrast to two ortho‐positioned dimethylamino groups).Crystals of 2 and 4. The energy difference was calculated to be 32 kJ mol−1 in favour of 4. Figure 2 and 4. For both molecules, the HOMO is localised primarily on the aromatic core. The relatively high HOMO energies (−3.93 eV for 2 and −4.05 eV for 4) are in line with the observed electron donor character (see below). In the case of the HOMO‐1 orbitals, the isodensity plots exhibit for both molecules one nodal plane but show a different electron‐density distribution. For 2, the nodal plane intersects the three parallel C−C bond axes of the naphthalene core. In the case of 4, the nodal plane intersects only the central C−C bond and includes two diagonal C atoms of the naphthalene core. These differences also affect the structures of the dicationic molecules (see discussion below).Quantum chemical calculations using the B3LYP functional in combination with the def2‐TZVP basis set were carried out to compare the relative energy of isomers 5  versus ferrocenium/ferrocene (Fc+/Fc) for the redox couple 5/52+ in CH2Cl2. For compound 4 the two‐electron oxidation and reduction waves seem to be slightly split, arguing for two one‐electron steps at potentials of E1/2=−0.51 V (4/4.+) and E1/2=−0.40 V (4.+/42+). Hence, the new compounds are slightly weaker electron donors than compounds 2 or 3 (E1/2=−0.65 V vs. Fc+/Fc for 2/22+ and E1/2=−0.71 V vs. Fc+/Fc for 3/32+).2 has the more negative redox potential and the higher HOMO energy compared with that of 4 , two more (reversible) one‐electron oxidation steps are visible. These are assigned to the redox couples 42+/4.3+  and 52+/5.3+ , as well as the redox couples 4.3+/44+  and 5.3+/54+ . In addition, broad shoulders towards higher potentials of the last oxidation step generating the tetracation are visible. As shown in the next section, the tetracation is barely soluble in standard organic solvents and is extremely reactive. Therefore, these additional features might arise from segregation effects at the electrode and/or decomposition of the highly reactive tri‐ or tetracationic compound. This would also explain the relatively low reduction currents.Cyclic voltammetry (CV) measurements of compound 5 Figure  found onEox=−0.4 V versus52+ in CH2Cl2 solution (see Supporting Information). Electronic excitation energies in the visible region signal the presence of an extended π‐conjugated system but removal of aromaticity. For comparison, upon two‐electron oxidation, compound 1 shows strong absorptions near 430 nm. The lower electronic excitation energy of 52+ compared with that of 12+ could be rationalised by the larger π‐conjugated system in 52+.The combination of electrochemical oxidation with UV/Vis spectroscopy (spectro‐electrochemistry) was used to obtain information about the oxidised species. These measurements show that the two‐fold oxidised compounds exhibit broad absorptions in the Vis/NIR region, with a maximum of absorption at 778 nm for compound 5 chemically by reaction with two equivalents of NO(BF4). Indeed, neutral 5 was oxidised cleanly to the dication 52+, and the product salt 5(BF4)2 was isolated in 84 % yield. In Figure 5 (reduced form) and 5(BF4)2, dissolved in CH2Cl2, are compared. Oxidation leads to the appearance of a broad absorption in the visible region (λmax=778 nm) and a small absorption around λmax=458 nm, in line with the results from spectro‐electrochemical measurements. In combination with strong absorptions centred below 400 nm that extend into the visible region, a deep‐green colour results.Motivated by the reversibility of the redox processes in the CV experiments, we next tried to oxidise compound 5(BF4)2 suitable for a structural analysis by single‐crystal X‐ray diffraction (XRD) measurements. Therefore, we repeated the oxidation of 5 with Ag(SbF6) and were able to isolate suitable crystals of the product 5(SbF6)2 in 88 % yield 2 are compared. The variations in the C−C bond lengths within the central ring system indeed signal loss of aromaticity. Some of the C−C bond lengths are significantly elongated upon oxidation. For example, the C1−C2 bond length increases from 1.396(2) Å before oxidation to 1.475(2) Å after oxidation. The C1−N1 bond length decreases to 1.326(2) Å, and the C2−N4 bond length decreases to 1.355(2) Å. The structure is in line with the Lewis representation drawn in Scheme 3 planes of each guanidino group are highly twisted to the central ring plane, C1‐N1‐C6 and C2‐N4‐C11 bond angles of 119.8(8) and 126.7(9)°, respectively. In this conformation, the N1/N4 atoms could establish a π‐bond with the C1/C2 atoms and at the same time establish some π‐interactions with the C6/C11 atoms.It was not possible to grow crystals of ith AgSbF and were5 with four equivalents of Ag(SbF6) in dichloromethane led to a purple precipitate insoluble in dichloromethane and 1,2‐difluorobenzene. Attempts to dissolve the possible tetracation in acetonitrile, diethyl ether, tetrahydrofuran or dimethylformamide resulted in a green solution indicating reduction to the dication, as confirmed through NMR spectroscopy. Given that all our attempts to dissolve 5(SbF6)4 without conversion failed, it was not possible to isolate a pure compound.Preparative oxidation of 5 with N‐halogenosuccinimides (NXS) or N‐bromophthalimide (NBP) did not only lead to halogeno‐substituted compounds but also to selective substitution by succinimido/phthalimido groups. After reduction with hydrazine, the neutral 1,2,5,6‐tetrakis(dimethylethyleneguanidino)‐3,7‐dihalogeno‐4,8‐disuccinimido‐naphthalenes (6 and 7) and 1,2,5,6‐tetrakis(dimethylethyleneguanidino)‐3,7‐dibromo‐4,8‐diphthalimido‐naphthalene (8) were obtained in good yields 2 with two equivalents of potassium phtalimide but observed no conversion. This result means that phthalimide substitution is possible only after substrate oxidation, given that it does not occur from the neutral form of the substrate. According to these results, we propose a multiple‐step pathway, starting with the oxidation of 5 to either 5.3+ or 54+ by two equivalents of the used reagent, followed by substitution at the positions 4 and 8 by the produced succinimide/phthalimide anion and deprotonation. The resulting disubstituted compound is again oxidised by two more equivalents of the reagent, substituted by the corresponding halides at positions 3 and 7, and deprotonated. Final reduction with hydrazine gives the neutral end‐product. The preference for halide over succinimide/phthalimide substitution in the last substitution steps is presumably due to steric effects.In a preliminary effort to rationalise the pathway of these intriguing reactions, we carried out experiments with different amounts of 2Cl2 solutions were also carried out for compounds 6–8 , −0.25 V (Eox=−0.18) and Eox=−0.18 V vs. Fc+/Fc in CH2Cl2 for 6, 8 and 7, respectively). Similar to the free ligands 4 and 5, for all compounds two more (reversible) one‐electron oxidation steps were visible at higher potentials (see Supporting Information). These are assigned to the redox couples 2+/.3+ and .3+/4+. For all three compounds, the redox couples 2+/.3+ as well as the redox couples .3+/4+ are (as expected) shifted to higher potentials in comparison to that of 5 (see Table CV measurements in CH4 and 5 could easily be doubly protonated to give (4+2 H)(PF6)2 and (5+2 H)(PF6)2 by reaction with two equivalents of NH4PF6. With an excess of HCl⋅OEt2, they are tetra‐protonated to the salts (4+4 H)Cl4 and (5+4 H)Cl4. The bands in the electronic absorption spectra experience small hypsochromic shifts upon protonation. Hypsochromic shifts upon protonation have been reported previously for guanidines.The new GFAs λ=471 nm for (5+2 H)(PF6)2 and λ=463 nm for (5+4 H)Cl4 (PF6)2 (λex=300 nm) and 15 % for (5+4 H)Cl4 (λex=280 nm). The drastic increase in the fluorescence intensity with the degree of protonation becomes evident from the photos in Figure 5 does not display ESPT, given that changes in the degree of protonation do not lead to shifts in the energy of the fluorescence signal.In clear difference to all other GFA compounds, blue fluorescence is switched on by protonation. Although the neutral compounds are fluorescence silent, the protonated compounds show fluorescence signals, with maxima of emission at 4 Figure . The qua6)2 λex=0 nm and 1H NMR spectrum of (5+2 H)(PF6)2 in CD3CN, two of the guanidino methyl groups experience a strong shift relative to 5, indicating a localised protonation  and the absence of N−H⋅⋅⋅N hydrogen bonds. By contrast, the 1H NMR spectrum of (2+2 H)(PF6) shows a singlet for all methyl protons of the four guanidino groups, indicating delocalised bonding of the protons in N−H⋅⋅⋅N hydrogen bonds, and (3+2 H)2+ certainly also exhibits this bonding 2+, also indicating protonation of two guanidino groups with terminal N−H bonds in the solid state. In contrast to the two‐fold protonated forms of 2 and 3, N−H⋅⋅⋅N hydrogen bonds are absent in the protonated compounds 4 and 5. Hence, they are not proton sponges.In the 5 was reacted with two equivalents of ZnCl2 or Pd(OAc)2, leading to the dinuclear complexes [5(ZnCl2)2] and [5{Pd(OAc)2}2] (see Scheme 5{Pd(OAc)2}2] were obtained through diffusion of diethyl ether in a saturated dichloromethane solution 2], the cobalt atoms are 1.034 Å above and below this plane.5{Pd(OAc)2}2] remain in the naphthalene ring plane. The differences in the coordination geometries can be rationalised by the different orientations of the lone‐pair imino N orbitals in 2/3 and 4/5.Next, compound 5(ZnCl2)2] is an irreversible process 2] compared to Eox=−0.43 V for free 5). As in free 5, two electrons are removed at equal potential. Interestingly, the reduction seems to occur in two potentially separated waves. Overall, the redox processes are not reversible, indicating some kind of transformation of the oxidised complex .CV measurements show that oxidation of [s Figure . Complex5{Pd(OAc)2}2] were studied in CV measurements  for the redox couple [5{Pd(OAc)2}2].+/[5{Pd(OAc)2}2] and E1/2=−0.01 V (Eox=0.04 V) for the redox couple [5{Pd(OAc)2}2]2+/[5{Pd(OAc)2}2].+. By using the formulas in Equations Next, the redox properties of .+ can be estimated to be 29 kJ mol−1 and the equilibrium constant Kdisp to be 8.49×10−6 (F=Faraday constant). Hence, by complexation three stable redox states of 5  are assessible. We showed previously that complexation of 1 with metal acetate leads in some cases to two one‐electron waves, allowing the isolation of dinuclear complexes bearing a radical monocationic or dicationic GFA ligand.(with 5{Pd(OAc)2}2] show a clear UV/Vis band around 780 nm upon removal of two electrons, confirming the presence of a dicationic ligand unit. When the oxidation–reduction cycles were repeated several times, the absorption maintained its intensity showing that the ligand‐centred redox process is fully reversible.Preliminary spectro‐electrochemical measurements for , tin(II) chloride, 2,2′‐bis(diphenylphosphino)‐1,1′‐binaphthyl (BINAP) and benzophenone imine were purchased from abcr, Sigma–Aldrich or Alfa Aesar and used without further purification. The reagents 2,6‐dibromo‐1,5‐dinitronaphthalene,1H NMR spectra and most of the 13C NMR spectra measured are very dilute. As a result, some of the aromatic 13C signals could not be detected and therefore could not be assigned. Elemental analyses were performed at the Microanalytical Laboratory of the University of Heidelberg by using the vario EL and vario MICRO cube devices from Elementar Analysensysteme GmbH. Owing to the high sensitivity of the synthesised compounds to oxygen and water, some elemental analyses deviate by more than 0.4 %. ESI mass spectrometry was performed with a JEOL JMS‐700 magnetic sector and BRUKER ApexQe hybrid 9.4 T FT‐ICR apparatus at the MS Laboratory of the University of Heidelberg. UV/Vis spectra were measured with a VARIAN Cary 5000 UV‐VIS‐NIR spectrometer. CV measurements were carried out with an EG&G Princeton 273 apparatus with an Ag/AgCl reference electrode. All voltammograms were recorded at room temperature. CH2Cl2 was used as solvent for the individual compounds (c=10−3 m), and N(nBu)4(PF6) , Fluka) was employed as supporting electrolyte (c=0.1 m). Spectro‐electrochemical studies were recorded with an AvaSpec 2048×14 CCD spectrometer and a Metrohm Autolab PGSTAT 204 potentiostat/galvanostat in an RHD Instruments TSC spectro sealable cell . Fluorescence measurements were recorded with a Varian Cary Eclipse instrument. Quantum yields were estimated with a PTI Quantum Master 40 equipped with an Ulbricht sphere.All reactions were carried out under a dry argon atmosphere using standard Schlenk techniques or in a dinitrogen‐filled glove box . The applied solvents were dried with an MBraun Solvent Purification System and degassed prior to use. 2,6‐Dibromonaphthalene, sodium‐N,N′‐bis: Sodium‐tert‐butyloxide , BINAP  and Pd2(dba)3  were dissolved in degassed toluene (40 mL). The deep‐red solution was stirred at room temperature for 10 min before 2,6‐dibromo‐1,5‐dinitronaphthalene  was added in portions. After stirring for another 5 min, benzophenone imine  was added dropwise, leading to precipitation of a brown solid. The reaction mixture was stirred for 3 h at 80 °C. Then the solution was allowed to cool down to room temperature, and the solvent was removed under vacuum. The obtained dark‐brown solid was washed with absolute methanol (3×15 mL) and dried under vacuum to yield 1.52 g  product as a brown powder. C, H, N analysis (%) for C36H24N4O4: calcd C 74.99, H 4.20, N 9.72; found C 74.24, H 4.52, N 9.23. 1H NMR : δ=7.61 , 7.33–7.56 , 6.74 ppm .1,2,5,6‐Tetraamino‐naphthalene tetrahydrochloride: N,N′‐bis  and SnCl2  were dissolved in concentrated HCl (30 mL). The red solution was stirred at 70 °C for 3 h. After cooling down the solution to 5 °C, the precipitated red‐brown product was filtered off and washed with degassed EtOH until the washing solution turned colourless. The product was obtained as a white‐grey solid in 95 % yield . 1H NMR : δ=7.51 , 7.24 ppm .1,2,5,6‐Tetrakis(tetramethylguanidino)‐naphthalene (4): 1,2,5,6‐Tetraamino‐naphthalene tetrahydrochloride  was suspended in degassed acetonitrile (15 mL) at 0 °C. A solution of 2‐chloro‐1,1′,3,3′‐tetramethylformamidinium chloride  in acetonitrile (5 mL) was added dropwise to the resulting grey suspension. The reaction mixture was stirred for 10 min at room temperature. Upon addition of triethylamine , the reaction mixture changed colour to yellow. The solution was stirred for 1 h at 0 °C and an additional 2 h at room temperature. After removing the solvent from the brown solution under vacuum, the residue was redissolved in 25 % NaOH solution and extracted with dichloromethane (3×20 mL). The combined organic phases were dried over potassium carbonate, and the solvent was removed under vacuum. The crude product was recrystallised from acetonitrile to yield 80 mg  of a colourless solid. Crystals suitable for structural characterisation by single‐crystal X‐ray diffraction were grown by diffusion of diethyl ether into a saturated acetonitrile solution. C, H, N analysis (%) for C30H52N12: calcd C 62.04, H 9.02, N 28.94; found C 61.60, H 8.90, N 28.53. 1H NMR : δ=7.21 , 6.70 , 2.64 ppm . 1H NMR : δ=7.12 , 6.54 , 2.64 ppm . 13C NMR : δ=122.60 , 116.32 , 39.90 , 39.49 ppm . MS (ESI+): m/z (%)=581.4 (100) [M+H]+, 536.4 (18) [M−NMe2]+, 491.3 (3) [M−(NMe2)2]+. UV/Vis (CH3CN): λmax (ϵ in L mol−1 cm−1)=257 (2.49×104), 348 nm (0.62×104). IR (KBr): ṽ=2924w, 2804w, 1617vs, 1600vs, 1560s, 1371m, 1307w, 1325w, 1136m, 990w, 898w, 613m, 480w cm−1.1,2,5,6‐Tetrakis(dimethylethyleneguanidino)‐naphthalene (5): 1,2,5,6‐Tetraamino‐naphthalene tetrahydrochloride  was suspended in degassed acetonitrile (15 mL) at 0 °C. Then 2‐chloro‐1,3‐dimethylimidazolinium chloride  in acetonitrile (5 mL) was added dropwise to the resulting grey suspension, and the reaction mixture was stirred for 15 min at room temperature. Triethylamine  was added, resulting in a colour change to red. Subsequently, the solution was stirred for 1 h at 0 °C and for an additional 2 h at room temperature. After removing the solvent from the red solution, the residue was redissolved in 25 % NaOH solution and extracted with dichloromethane (3×20 mL). The combined organic phases were dried over potassium carbonate, and the solvent was removed under vacuum. The crude product was recrystallised from acetonitrile to yield 135 mg  of a colourless solid. Crystals suitable for an X‐ray analysis were grown from a saturated acetonitrile solution. C, H, N analysis (%) for C30H44N12: calcd C 62.91, H 7.74, N 29.35; found C 62.18, H 7.12, N 29.89. 1H NMR : δ=7.30 , 6.81 , 3.20 , 2.58 ppm . 13C NMR : δ=122.60 , 116.56 , 49.18 , 49.07 , 35.23 , 34.59 ppm . MS (ESI+): m/z (%)=573.4 (100) [M+H]+. UV/Vis (CH2Cl2): λmax (ϵ in L mol−1 cm−1)=276 (2.19×104), 340 nm (0.56×104). IR (KBr): ṽ=2932w, 2842w, 1660vs, 1628vs, 1561s, 1485m, 1394m, 1275m, 1037m, 965w, 910s, 827m, 763w, 712w cm−1.4)2Compound 5 was dissolved in THF (5 mL). Then NO(BF4)  was added in one portion. The solution was stirred for 2 h at room temperature. During this time, a green solid precipitated. The crude product was filtered off and washed with THF (3×3 mL) and Et2O (3×3 mL). The obtained solid was dried under vacuum to yield 22 mg  of the product as a green solid. C, H, N analysis (%) for C30H44N12B2F8: calcd C 48.28, H 5.94, N 22.25; found C 48.30, H 6.27, N 22.10. 1H NMR : δ=7.77 , 6.00 , 3.79 , 3.59 , 2.80 , 2.76 ppm . 13C NMR : δ=48.73 , 48.24 , 33.45 , 33.30 ppm . UV/Vis (CH2Cl2): λmax (ϵ in L mol−1 cm−1)=230 (3.29×104), 277 (3.22×104), 778 nm (0.23×104).6)2Compound 5 was dissolved in dichloromethane (5 mL). Then Ag(SbF6)  was added in one portion. The solution was stirred for 2 h at room temperature. The dark‐green solution was filtered from the precipitated silver, and the solvent was removed under vacuum. The crude product was washed with THF (3×3 mL) and Et2O (3×3 mL). The obtained solid was dried under vacuum to yield 32 mg  of the product as a green solid. Crystals suitable for structural characterisation by single‐crystal X‐ray diffraction were grown by diffusion of pentane into a saturated dichloromethane solution. C, H, N analysis (%) for C30H44N12Sb2F12: calcd C 34.51, H 4.25, N 16.10; found C 34.89, H 4.52, N 16.13. 1H NMR : δ=7.77 , 6.00 , 3.79 , 3.59 , 2.80 , 2.76 ppm . 13C NMR : δ=48.73 , 48.24 , 33.45 , 33.30 ppm .6)2Compound (5+2 H) was dissolved in tetrahydrofuran (5 mL). Ammonium hexafluorophosphate  was added in one portion to the light‐yellow solution. Then the reaction mixture was stirred at room temperature for 1 h. During this time, a white solid precipitated. The crude product was filtered and washed with tetrahydrofuran until the solution turned colourless. Remaining tetrahydrofuran was removed under vacuum to give the product as a white solid in 87 % yield . Crystals of (5+2 H)(BF4)2 suitable for structural characterisation by single‐crystal X‐ray diffraction were grown by diffusion of diethyl ether into a saturated solution of 5(BF4)2 in dichloromethane. C, H, N analysis (%) for C30H46N12F12P2⋅THF: calcd C 43.59, H 5.81, N 17.94; found C 43.22, H 6.31, N 17.25. 1H NMR : δ=7.65 , 7.18 , 3.54 , 3.45 , 2.69 , 2.58 ppm . 13C NMR : 49.44 , 49.34 , 34.63 , 34.32 ppm . UV/Vis (CH3CN): λmax (ϵ in L mol−1 cm−1)=211 (3.05×104), 257 (2.51×104), 301 nm (2.10×104). IR (KBr): ṽ=3357m, 2947m, 2882m, 2848m, 1629s, 1604s, 1574m, 1489w, 1443w, 1423w, 1405m, 1390m, 1323w, 1299m, 1287s, 1242m, 1202w, 1142w, 1123w, 1040m, 1021m, 965w, 914m, 879w, 833vs, 787m, 766m, 742w, 712w, 677w cm−1.4Compound (5+4 H)Cl: Compound 5  was dissolved in tetrahydrofuran (5 mL). HCl  was added to the light‐yellow solution. After 1 h of stirring at room temperature, a white solid precipitated. The crude product was filtered and washed with tetrahydrofuran (3×5 mL) and diethyl ether (3×5 mL). Then the solid was dried under vacuum to give the product as a colourless solid in 95 % yield . The compound is extremely hygroscopic and quickly picks up H2O. C, H, N analysis (%) for C30H48N12Cl4⋅H2O: calcd C 48.92, H 6.84, N 22.28; found C 48.80, H 7.18, N 22.26. 1H NMR : δ=11.95 , 11.80 , 8.05 , 7.65 , 3.75 , 2.96 , 2.78 ppm . 13C NMR spectroscopic shifts are not given because of low solubility. UV/Vis (CH3CN): λmax (ϵ in L mol−1 cm−1)=268 (2.25×104), 304 nm (0.47×104). IR (KBr): ṽ=2930m, 2877m, 2801m, 2660m, 1591vs, 1479m, 1401m, 1373s, 1336w, 1296s, 1127w, 1080w, 1037m, 970m, 913m, 834w, 785w, 686m cm−1.N1,2,5,6‐Tetrakis‐3,7‐dibromo‐4,8‐disuccinimido‐naphthalene (6): Compound 5  was dissolved in dichloromethane (5 mL). Then N‐bromosuccinimide  was added in one portion. The colour of the solution immediately changed to deep yellow. After 1 h of stirring at room temperature, the colour changed to green. The solvent was removed under vacuum, and the remaining solid was washed with tetrahydrofuran (2×5 mL). The oxidised product was dissolved in dichloromethane (8 mL), and hydrazine hydrate  was added. Gas development and a colour change to light yellow were observed. Subsequently aqueous NaOH solution  was added, and the mixture was stirred for an additional 20 min. The organic phase was separated and dried over MgSO4. The product was obtained, after solvent removal, as a yellow solid in 69 % yield . Crystals suitable for an X‐ray analysis were grown by diffusion of diethyl ether in a saturated dichloromethane solution. C, H, N analysis (%) for C38H48N14Br2O4: calcd C 49.36, H 5.23, N 21.21; found C 50.01, H 5.72, N 20.81. 1H NMR : δ=3.15 , 2.76 , 2.59 , 2.45 ppm . 13C NMR : δ=176.73 , 49.02 , 48.69 , 34.75 , 33.98 , 29.35 ppm . MS (ESI+): m/z (%)=925 (100) [M+H]+, 846 (20) [M−Br]+. UV/Vis (CH2Cl2): λmax (ϵ in L mol−1 cm−1)=230 (3.69×104), 293 (4.81×104), 355 (0.88×104), 420 nm (1.07×104). IR (KBr): ṽ=2935m, 2846m, 1715vs, 1648vs, 1633vs, 1536w, 1489m, 1441w, 1420s, 1387s, 1338m, 1280s, 1243s, 1190s, 1114w, 1071w, 1045m, 1028m, 998w, 965w, 933s, 828m, 797m, 765m, 721m, 710m, 705m, 697m cm−1.N1,2,5,6‐Tetrakis‐3,7‐diiodo‐4,8‐disuccinimido‐naphthalene (7): Compound 5  was dissolved in dichloromethane (5 mL). Then N‐iodosuccinimide  was added in one portion. The colour of the solution immediately changed to deep yellow. After 2 h of stirring at room temperature, the colour turned green. The solvent was removed under vacuum, and the remaining solid was washed with tetrahydrofuran (2×5 mL). The oxidised product was dissolved in dichloromethane (8 mL), and hydrazine hydrate  was added. Gas development and a colour change to light yellow were detected. Subsequently aqueous NaOH solution  was added, and the mixture was stirred for an additional 20 min. The organic phase was separated and dried over MgSO4. The product was obtained, after solvent removal, as a yellow solid in 65 % yield . Crystals suitable for an X‐ray analysis were grown by diffusion of diethyl ether in a saturated dichloromethane solution. C, H, N analysis (%) for C38H48N14I2O4: calcd C 44.80, H 4.75, N 19.25; found C 44.94, H 5.66, N 19.03. 1H NMR  δ=3.15 , 2.76 , 2.60 , 2.44 ppm . 13C NMR : δ=176.67 , 49.01 , 48.68 , 34.76 , 33.99 , 29.53 ppm . MS (ESI+): m/z (%)=1019 (45) [M+H]+, 893 (100) [M−I]+, 767 (25) [M−2I]+. UV/Vis (CH2Cl2): λmax (ϵ in L mol−1 cm−1)=231 (2.98×104), 295 (3.47×104), 359 (0.62×104), 416 nm (0.69×104). IR (KBr): ṽ=3002w, 2927m, 2873m, 2798w, 1721m, 1570s, 1497m, 1495s, 1424m, 1370vs, 1299s, 1234m, 1179s, 1132s, 1064m, 1026m, 1004m, 946w, 918w, 906w, 895w, 850w cm−1.N1,2,5,6‐Tetrakis‐3,7‐dibromo‐4,8‐diphthalimide‐naphthalene (8): Compound 5  was dissolved in dichloromethane (5 mL). Then N‐bromophthalimide  was added in one portion. The colour of the solution immediately changed to green. After 2 h of stirring at room temperature, the solvent was removed under vacuum, and the resulting solid was washed with tetrahydrofuran (2×5 mL). The oxidised product was dissolved in dichloromethane (8 mL), and hydrazine hydrate  was added. Gas development and a colour change to red were detected. Then aqueous NaOH solution  was added to the mixture and stirring continued for 20 min. The organic phase was separated and dried over MgSO4. The solvent was removed, giving the product as an orange‐red solid in 82 % yield . Crystals suitable for an X‐ray analysis were grown by diffusion of diethyl ether in a saturated dichloromethane solution. C, H, N analysis (%) for C46H48N14Br2O4: calcd C 54.13, H 4.75, N 19.21; found C 53.52, H 5.03, N 19.28. 1H NMR : δ=7.89 , 7.74 , 3.07 , 2.57 , 2.16 ppm . 13C NMR : δ=167.60 , 134.07 , 133.87 , 123.38 , 48.70 , 48.11 , 34.09 , 33.46 ppm . UV/Vis (CH2Cl2): λmax (ϵ in L mol−1 cm−1)=234 (4.56×104), 293 (1.77×104), 363 (0.78×104), 410 nm (0.93×104). IR (KBr): ṽ=2961w, 2921m, 2842m, 1713s, 1617vs, 1488m, 1440w, 1415m, 1388m, 1347w, 1317w, 1281m, 1253s, 1087m, 1039m, 1021s, 948m, 921w, 883m, 794s, 712vs cm−1.2)2]Compound +, 808 (7) [M−Cl]+, 709 (20) [M−ZnCl2]+, 573 (100) [M−(ZnCl2)2]+. UV/Vis (CH3CN): λmax (ϵ in L mol−1 cm−1)=234 (0.59×104), 288 (0.95×104), 346 (0.39×104), 398 nm (0.09×104). NMR spectra were not measured because of low solubility.2}2]Compound  and the intermediate of 7). Data were processed with the standard Nonius and Bruker  software packages.1968384, 1968385, 1968386, 1968387, 1968388, 1968389, 1968390, 1968391, 1968392, and 1968393 contain the supplementary crystallographic data for this paper. These data are provided free of charge by The Cambridge Crystallographic Data Centre.Full shells of intensity data were collected at low temperature with a Nonius Kappa CCD diffractometer  should be addressed to the authors.SupplementaryClick here for additional data file.


The highlights of this method are:  Specifications Tableα-Protected amino acids, Fmoc-Arg(Pdf)-OH, Fmoc-Leu-OH, Fmoc-Gly-OH, Fmoc-Pro-OH, Fmoc-Trp (Boc)-OH, Fmoc-d-Trp (Boc)-OH, 2-Cl-trityl resin (loading 0.85 mmol g_1) and Rink amide MBHA resin (loading 0.557 mmol g_1), were purchased from GL Biochem Ltd. . Fmoc-Ser(tBu)-OH, Fmoc-Lys (Boc)-OH, Fmoc-Phe-OH, Fmoc-His(Trt)-OH and N—Hydroxybenzotriazole (HOBt), were purchased from Nanjing Peptide Biotech Ltd. N, N-diisopropylcarbodiimide (DIC), trifluoroacetic acid (TFA), trifluoroethanol (TFE) and N, N-diisopropylethyl amine (DIPEA) were purchased from Energy chemical. Paclitaxel was purchased from Ark Pharm. Pteroic acid and 2-(7-Azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphatewas (HATU) purchased from Bide Pharmatech. Cystamine dihydrochloride was purchased from Aladdin. All other reagents, unless otherwise indicated, were obtained from Sigma-Aldrich Co. .All reagents were purchased as reagent grade and used without further purification. N. Synthesis is generally synthesized from the C-terminus (carboxy terminus) to the N-terminus (amino terminus). Since Merrifield developed a solid phase peptide synthesis method in 1963, it has been continuously improved and perfected . Solid phase method has become a common technique in peptide and protein synthesis and shows the advantages over classical liquid phase synthesis . From the first amino acid to the last solid-phase synthesis was used widely to prepare peptides. However, the method was limited because its purity has been declining as the synthesis progresses  .Peptide synthesis is a process of repeating the addition of amino acidsynthesis . From th peptides. HoweverA new strategy for solid phase peptide synthesis in the fragment synthesis based on the use of 2-Cl-trityl resin as carrier to obtain protected peptide-resin and Rink Amide Resin, HOBT/HBTU or PyBOP/DIPEA or HATU/DIPEA as the coupling method is described. In this study, we introduced a fragment synthesis method of target conjugate . The fraFragment 1 and Fragment 2 were synthesized using standard 9-fluorenylmethoxycarbonyl (Fmoc) solid phase synthesis techniques.Fragment 1 was prepared on 2-Cl-trityl resin. Synthetic route of Fragment 1 was shown in Progress one:1, Place the 2-Cl-trityl resin  in a glass funnel and swell the resin with DCM (30 ml) for 30 min; 2, Weigh out Fmoc-Gly-OH (5 eq), DIPEA (10 eq); 3, Add 15 mL of DCM to effect total dissolution and blow nitrogen; 4, Agitate resin gently for 1 h; 5, Deprotection 0.1 mmol/L HOBt, 20% Piperidine/DMF (V/V), two times; 6, Wash resin with DMF(4 × 20 ml); 7, Weigh out Fmoc-amino acid (5 eq), HOBT (5 eq) and HBTU (5 eq) and 18 mL of DMF to effect total dissolution, add DIPEA (10 eq) and blow nitrogen for 2 h; 8,Repeat steps 5–7; 9, Cleavage of protected peptide fragment from 2-Cl-trityl resin. Place 1 g peptide-resin in a flask, add 15 ml of TFE/DCM (2:8) cleavage solution for 45 min; 10. Concentrate the filtrates and add 20 ml cold ether and 10 ml hexane; 11.Centrifugate and collect the resulting solid and wash with ether (4 × 10 ml); 12. Dry the protected peptide in vacuum and store at −20 °C (Fragment 1).3  was dissolved in 2 ml H2O and 20 ml Dioxane. Succinic anhydride  was added to the above mixture in the ice bath. The resulting reaction mixture was stirred overnight. Then, 0.33 mL of DIPEA and  of Fmoc-OSu were added to the reaction mixture at 0 °C. The reaction mixture was stirred for 4 h. Then, the filtrate was concentrated under reduced pressure. 40 mL of water was added. the white precipitate was collected by filtration, and dried in vacuum, sequentially. 0.34 g of Fmoc-cystamine succinic acid (Fmoc-CS) was acquired by column chromatography  with a yield of 36%. Fmoc-CS, white solid; yield, 36%. 1H NMR  δ 8.14 ; δ 7.88–7.90 ; 7.68–7.70 ; 7.39–7.44 ; 7.31–7.35 ; 6.28 ;4.30–4.32 ; 4.22–4.23 ; 3.06–3.34 ; 2.50–2.76 ; 2.23–2.35 . MS (ESI) m/z calcd for [C32H26N2O5S2 + Na]+ 497.6, found 497.7.Progress two:Cystamine dihydrochloride  and NaHCOFragment 2 was prepared on Rink Amide MBHA resin. Synthetic route of Fragment 2 was shown in Progress three:1, Place the Rink Amide MBHA resin in a glass funnel and swell the resin with DCM (30 ml) for 30 min; 2, Deprotection: 0.1 mmol/L HOBt, 20% Piperidine/DMF (V/V), two times; 3, Wash resin with DMF(4 × 20 ml); 4. Weigh out Fmoc-amino acid (5 eq), HOBT (5 eq) and HBTU (5 eq); 5, Add 18 mL DMF to effect total dissolution; 6, Add DIPEA (10 eq) and blow nitrogen; 7, Agitate resin gently for 2 h; 8, Repeat steps 2–6; 9, Collect the peptide-resin (Fragment 2).Synthetic route and MS data of target conjugate was shown in 2O: TFA (2.5:2.5:95). 11, Concentrate the filtrates and add 40 ml cold ether, collect the solid and wash the solid with cold ether four times; 12, Dry the target peptide in vacuum and store at −20 °C.Progress four:1, Place the Fragment 2 into a glass funnel and swell the resin with DCM(30 mL) for 30 min; 2, Add the Linker (2 eq), PyBOP (3 eq) and DIPEA (3 eq); 3, Add 16 mL of DMF to effect total dissolution and blow nitrogen for 4 h; 4, Wash resin with DMF(3 × 20 ml); 5. Deprotection: 0.1 mmol/L HOBt, 20% Piperidine/DMF (V/V), two times; 6. wash resin with DMF(4 × 20 ml); 7, Add Fragment 1(2 eq), HATU (2eq) and DIPEA(4 eq); 8, Add 14 mL of DMF to effect total dissolution and blow nitrogen for 4 h; 9. wash resin with DMF(3 × 20 ml) and DCM(2 × 20 ml); 10, Cleavage of target conjugate: TIS: HThe authors have no conflicts of interest to declare.


Novozym® 435 showed sn-1 regiospecificity to phosphatidylcholine, although it does not exhibit regiospecificity to triacylglycerol. In particular, quantitative 1-LPC yields (96.5 ± 0.2 mol%) were reliably obtained in the presence of Novozym® 435 (100 wt% of PC), 97 % ethanol, in 72 h at 40 °C. During the reaction, acyl migration from 1-LPC to 2-LPC was rare. This novel synthetic method is expected to expand the practical applications of 1-LPC.Efficient preparation methods for 1-lysophosphatidylcholine (1-LPC), a physiologically important compound, are lacking. Here, we established a method for 1-LPC preparation via Novozym® 435 (a lipase B from  It exerts a variety of physiological functions; for example, it induces MCP-1 expression, increases inflammatory cytokines, disrupts mitochondrial integrity, activates macrophages, and induces oligodendrocyte demyelination [Lysophosphatidylcholine (LPC) is a class of biomolecules derived from the cleavage of phosphatidylcholine (PC) via phospholipase A2 (PLA2) . LPC haslination ,5. Howev2-catalyzed hydrolysis of PC is a simple and well-established method [sn-2 positional regiospecificity of PLA2 enables 2-LPC production in slightly alkali reaction conditions with calcium cations as co-activators of PLA2. This technique is often used to analyze the fatty acid composition at the sn-1, -2 positions of phospholipids [Candida antarctica) as a catalyst in a mixture of 50 % t-butanol and 0.5 mmol water at 25 °C for 100 h [Rhizomucor miehei) as a catalyst in a solvent-free system at 50 °C for 24 h (or 40 °C for 72 h) [To prepare 2-LPC, PLAd method ,7. The sholipids . The lipholipids . Carmen or 100 h . Mnasri or 72 h) .1 (PLA1)-catalyzed hydrolysis has been applied by Lim et al. [1 (Thermomyces lanuginosus)-catalyzed hydrolysis under a bi-phasic system of water and hexane [1 is limited by the high cost. In addition, quantitative yields have not been obtained using sn-1, 3 regiospecific lipase. 1-LPC production via lipase-catalyzed alcoholysis, first developed in 1994 by Sarney et al., has been demonstrated using Lipozyme® IM-60 (Mucor miehei) as a catalyst [1 (Thermomyces lanuginosus), Novozym® 435, and Lipase PS (Burkholderia cepacia), were reported by Baeza-Jiménez et al. [sn-1 positional specificity to glycerophospholipids but no regiospecificity to triacylglycerols [To prepare 1-LPC, phospholipase Am et al. . They obd hexane . Howevercatalyst ,14. Quancatalyst . In 2013z et al.  with LPCIn this study, a simple preparation method of 1-LPC via Novozym® 435-catalyzed ethanolysis without any toxic solvents was demonstrated  . The ef22.1Candida antarctica) was purchased from MIK Pharm Co., Ltd. . LPC (l-α-lysophosphatidylcholine) from egg yolk, used as the 2-LPC standard, was obtained from FUJIFILM Wako Pure Chemical Corporation .PC from soybean (Phospholipon 90 G) was purchased from H. Holstein Co., Ltd. . Novozym® 435  was dissolved with 1 mL of 97 % ethanol, followed by the addition of Novozym® 435 (100 wt% of PC). The reaction was allowed to proceed at 40 °C and 900 rpm in the dark. After the reaction, the enzyme was removed by filtration. The reaction mixture was adjusted to 5 mL with ethanol containing 0.1 % formic acid to avoid acyl migration and subjected to high-performance liquid chromatography (HPLC) for calculation of the 1-LPC yield.2.32  column was used. Samples were then eluted by isocratic elution of the mobile phase at 95 % ethanol- and 20 mM oxalic acid with a ratio of 94:4 (v/v). The flow rate was maintained at 1.0 mL/min and the column temperature was maintained at 25 °C. Calibration curves were prepared with substrate PC and 2-LPC . Statistical differences were determined using Scheffe’s tests, with a significance threshold of P < 0.05.33.1We first investigated the effect of the ethanol concentration on the ethanolysis reaction mediated by Novozym® 435. The reaction mixture included PC (50 μmol), solvents with different concentrations of ethanol (1.0 mL), and Novozym® 435 (100 wt% of PC). The reaction proceeded for 24 h at 60 °C and 900 rpm in the dark.sn-2 position did not occur in this reaction system. At high concentrations of ethanol, the 1-LPC yield was also low; 11.2 ± 5.5 mol% 1-LPC was obtained in 99.5 % ethanol. This may be explained by enzyme denaturation caused by ethanol, which can absorb essential water [The highest average yield (75.1 ± 1.3 mol%) was obtained using 97 % ethanol as a reaction solvent, although no significance differences in yield were observed for ethanol concentrations in the range of 92%–98% . The yieal water , as smalal water .Fig. 1EfBased on these results, 97 % ethanol was selected as the reaction solvent in the reaction system.3.2An ethanolysis reaction was performed under different temperatures. The reaction mixture consisted of PC (50 μmol), 97 % ethanol (1.0 mL), and Novozym® 435 (100 wt% of PC). The reaction proceeded for 24 h at 4–60 °C at 900 rpm in the dark.The highest 1-LPC yields were obtained at 40 °C (80.9 ± 8.7 mol%), although the yields at this temperature did not differ significantly from those at 60 °C . At 20 °Baeza-Jiménez et al. also investigated the effect of temperature on the Novozym® 435-catalyzed ethanolysis of PC, showing that the degree of ethanolysis increased clearly as the reaction temperature increased from 40 °C to 60 °C . They usThe 1-LPC yield did not differ significantly between reaction temperatures of 40 °C and 60 °C; accordingly, 40 °C was selected as the optimal reaction temperature in this system to avoid acyl migration at high temperatures.3.3Next, the effect of enzyme dose on 1-LPC yield and residual PC was investigated. The reaction mixture consisted of PC (50 μmol), 97 % ethanol (1.0 mL), and different doses of Novozym® 435 (12.5–100 wt% of PC). The reaction proceeded for 24 h at 40 °C, 900 rpm in the dark.1-LPC yield decreased with decreasing enzyme dose . In accoThis result indicated that this reaction system requires a substantial enzyme dose to reliably obtain high 1-LPC yield, likely because short chain alcohols, such as methanol and ethanol, tend to dehydrate enzyme protein and thereby deactivate the enzyme . Althoug3.4Finally, the effect of reaction time was investigated using a mixture consisting of PC (50 μmol), 97 % ethanol (1.0 mL), and Novozym® 435 (100 wt% of PC). The reaction was continued for 72 h at 40 °C, 900 rpm in the dark.1-LPC yield increased as the reaction time increased and reached a plateau at 60 h . The maxAlternatively, Baeza-Jiménez et al. reported that the degree of ethanolysis increases until 48 h and apparently decreases thereafter at several reaction temperatures (30–60 °C) . We note4sn-1 position of PC and acyl migration from 1-LPC to 2-LPC was rare.1-LPC was successfully prepared under the following optimized reaction conditions: PC (50 μmol), 97 % ethanol (1.0 mL), and Novozym® 435 (100 wt% of PC), with a 72-h reaction time at 40 °C and 900 rpm in the dark. This resulted in a 1-LPC yield of 96.5 ± 0.2 mol%. In this reaction system, Novozym® 435 showed regiospecificity at the Sayumi Yasuda: Investigation. Yukihiro Yamamoto: Conceptualization, Methodology, Writing - original draft.None.


In the crystal, O—H⋯S hydrogen bonds between the coordinating water molecules and thiocyanate groups form a supramolecular chain with a zigzag-shaped calcium skeleton.A new title heterometallic Cu/Ca complex, , has been synthesized using a one-pot reaction of copper powder, calcium oxide, o-vanillin and ammonium thiocyanate in methanol under ambient conditions. The Schiff base ligand (C8H9NO2) is generated in situ from the condensation of o-vanillin and ammonia, which is released from the initial NH4SCN. The title compound consists of a discrete binuclear molecule with a {Cu(μ-O)2Ca} core, in which the Cu⋯Ca distance is 3.4275 (6) Å. The coordination geometries of the four-coordinate copper atom in the [CuN2O2] chromophore and the seven-coordinate calcium atom in the [CaO5N2] chromophore can be described as distorted square planar and pentagonal bipyramidal, respectively. In the crystal, O—H⋯S hydrogen bonds between the coordinating water molecules and thiocyanate groups form a supramolecular chain with a zigzag-shaped calcium skeleton.The new heterometallic complex, aqua-1κ The copper atom is four-coordinated by two imino N and two phenoxo O atoms from the Schiff base ligands. The coordination geometry of the CuN2O2 chromophore is slightly distorted square planar; the Cu—O and Cu—N bond lengths vary in the range of 1.918 (2)–1.937 (2) Å and the corresponding cis/trans bond angles deviate from ideal symmetry by less than 8° with τ4 = 0.112 2} moiety chelate the calcium atom in a tetradentate manner and the coordination sphere of the Ca center is further completed by two SCN groups and one water molecule giving a coordination number of seven. The CaO5N2 chromophore can be described as having a distorted pentagonal–bipyramidal geometry with the oxygen atoms in the equatorial plane and the nitrogen atoms in the axial positions  and 2.521 (2) Å] are observed for the coordinating methoxy groups and the shortest ones [2.339 (2)–2.356 (2) Å] for the phenoxido groups and the water molecule. The values are in accordance with those found in related binuclear Cu/Ca complexes 2(H2O)] 2] 2Na(ClO4)(H2O)]  ring  link neighbouring chains, increasing the whole dimensionality of the crystal framework.The coordinating water molecule and thiocyanate ions of each binuclear complex are involved in four O—H⋯S hydrogen bonds Table 1 with twois Fig. 5. Weak N—is Fig. 5 and a π–et al., 2016et al., 1985et al., 2010et al., 2011et al., 2012L)2Ca(NCS)2(H2O)] is the first molecular binuclear Cu/Ca complex with an asymmetric Schiff base ligand to have been characterized crystallographically.To date, the crystal structures of 72 complexes containing copper and calcium are known ,CuL)2Ca(NCS)2(H2O)] was obtained. Its formation can be described by the following scheme:and the heterometallic complex [Cu(0 + CaO + 2o-vanillin + 2NH4SCN + 1/2O2(air)→ [Cu(L)2Ca(NCS)2(H2O)] + 3H2O,CuL can be regarded as a product of the condensation of o-vanillin and NH3, which is released from NH4SCN in the basic environment.where the Schiff base Ho-vanillin  and NH4SCN  were added to 30 ml of methanol. The reaction mixture was stirred magnetically at 323–333 K for ca 5 h until the complete dissolution of the copper powder was observed. The solution was filtered and left for 1 d, and then light-orange crystals were formed. Yield: 0.26 g . Analysis calculated for CaCuC18N4H18O5S2: Ca 7.45, Cu 11.81, C 40.18, N 10.41, H 3.37, S 11.92. Found: Ca 8.1, Cu 11.2, C 36.5, N 10.1, H 3.2, S 11.4. FT–IR : 3349 vs, 3187 vs, 2942 s, 2076 vs, 1617vs, 1555 m, 1464 vs, 1386 s, 1318 s, 1245 s, 1225 vs, 1162 m, 1074 s, 1036 m, 948 m, 853 m, 823 m, 738 s, 652 m, 617 m, 571 m, 515 m, 469 m.Copper powder , CaO , 3 groups were placed in idealized positions (C—H = 0.93–0.96 Å) and constrained to ride on their parent atoms, with Uiso(H) = 1.2Ueq(C) for CH and 1.5Ueq(C) for CH3. All H atoms of the NH and OH groups were located in a difference-Fourier map and refined isotropically; the N—H and O—H distances were restrained to have fixed lengths of 0.82 (1) and 0.85 (1) Å, respectively.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S205698902000211X/is5531sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S205698902000211X/is5531Isup2.hklStructure factors: contains datablock(s) I. DOI: 1984001CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Vitamins K and D are present in the human brain and have been implicated in Alzheimer’s disease and related dementias (ADRD). Because the use of banked brain tissue in ADRD research is increasing, we evaluated the stability of vitamin K and vitamin D in human brain tissue over long-term freezer storage using samples obtained from the Rush Memory and Aging Project . Specimens were stored at -80□C until analyzed. Vitamin K  and vitamin D (25(OH)D) were measured in four regions  and averaged across regions. Storage time was categorized into two-year increments. Differences in MK4 and 25(OH)D concentrations according to storage time were evaluated using general linear models. MK4 concentrations did not differ in brains stored ≤8 years . MK4 in brains stored >8.0 years (0.8±0.1 pmol/g) was 33% lower than the concentration in brains stored ≤2.0 years (p=0.005). The 25(OH)D concentrations did not differ in brains stored ≤6 years (geometric mean±SEM 25(OH)D pmol/g: storage ≤2.0 years=1.2±0.1, 2.1-4.0 years=1.1±0.1, 4.1-6.0 years=1.2±0.1; p≥0.37). The 25(OH)D concentration in brains stored >6.0 years was 31-37% lower than that in brains stored ≤2.0 years . MK4 and 25(OH)D appeared to be stable in human brain tissue stored at -80oC for up to 8 and 6 years, respectively. Storage time merits consideration when designing and interpreting studies that relate brain nutrient concentrations to ADRD.


The title compound is a demonstration of hydroboration of ynamines with borandiol ester. The bond lengths of the resulting push–pull olefin are discussed. N-[(Z)-2--2-phenylethenyl]-N-(propan-2-yl)aniline, C23H22BNO2, contains a C=C bond that is conjugated with a donor and an acceptor group. An analysis that included similar push–pull olefins revealed that bond lengths in their B—C=C—N core units correlate with the perceived acceptor and donor strength of the groups. The two phenyl groups in the molecule are rotated with respect to the plane that contains the BCCN atoms, and are close enough for significant π-stacking. Definite characterization of the title compound demonstrates, for the first time in a reliable way, that hydroboration of ynamines with borandiol esters is feasible. Compared to olefin hydroboration with borane, the ynamine substrate is activated enough to undergo reaction with the less active hydroboration reagent catecholborane. Boronic esters are frequently used to transfer organic groups to transition metals, for example in the transmetallation step of the Suzuki–Miyaura reaction. Hydroboration of ynamines with borandiol esters produces amino-functionalized boronic esters in one step and perfect atom economy.et al., 2000et al., 20016F5)2, can readily hydroborate l-propynyl-2,2,6,6-tetramethylpiperidine , the amine, and an electron-withdrawing group (EWG), the boronate. Therefore it belongs to the class of push–pull (captodative) olefins, which are known to have unusual properties such as weak π-bonds or biradical reactivity  = 2.771 Å and B1A⋯H5B = 2.856 Å . The shortest intermolecular distances between the catechol unit and boron are B1A⋯C21Bi = 3.540 (4) Å, B1A⋯C20Bi = 3.686 (4) Å , B1A⋯H21Bi = 3.113 Å and B1A⋯H20Bi = 3.381 Å . In addition there is a short contact between one of the other catechol hydrogen atoms and the meta-carbon of the aniline, H22B⋯C6B of 2.877 Å . All of these interactions involve atoms that are part of arenes and could be seen as intermolecular π-stacking.There is a short intermolecular contact between the boron atom and an aniline A⋯H2BA = 2.698 Å, O2B⋯H3AB = 2.631 Å, ΣrvdW = 2.70 Å] and with one of the anilic meta-H atoms  = 2.40 Å]. The nitrogen atom is sterically shielded by surrounding groups and does not have any close intermolecular neighbours.Methyl hydrogen atoms of the isopropyl group are at van der Waals distances with one of the oxygen atoms -N-(propan-2-yl)aniline: Under a counterflow of argon, 3.08 g of N-isopropyl-N-(phenylethynyl)aniline  were placed in an oven-dried 20 ml Schlenk flask with a Young valve. The flask and its contents were purged three times by applying high vacuum followed by flushing with argon. 6.5 ml dry tert-butyl methyl ether were added and the mixture stirred vigorously to ensure mixing of the two liquids. 2.1 ml of catecholborane  were added, the flask closed, and the reaction mixture heated to 323 K for 16 h. Cooling to room temperature led to the precipitation of the product. The supernatant was removed and the precipitate recrystallized from 40 ml of tert-butyl methyl ether. X-ray quality crystals were obtained in a yield of 1.88 g (40%). Notes: (a) Schlenk techniques are necessary because ynamines and catecholborane are moisture-sensitive; (b) the reaction also works well in diethyl ether, 1,4-dioxane or THF.1H NMR : δ (ppm) = 1.34 , 3.89 , 6.77–6.80 , 6.81–6.85 , 6.85–6.92 , 6.95–6.98 , 7.11–7.14 , 7.53 .13C1H NMR : δ (ppm) = 22.2 , 57.4 , 98.1 , 111.7 , 121.8 , 124.3 , 124.6 , 126.3 , 126.9 , 128.0 , 129.4 , 138.8 , 143.3 , 144.9 , 148.9 . Inverse gated {13}C{1}H} NMR with D1 = 60 s measured to get integrable 13C NMR.11B NMR : δ (ppm) = 33.2 (s).Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C-methyl). The absolute structure was not determined because of unreliable Flack and Hooft parameters.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989020005289/lh5953sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989020005289/lh5953Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989020005289/lh5953Isup3.cmlSupporting information file. DOI: 1997061CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


H)-one was crystallized as the hydrochloride salt 4--1-[(6-phenylpyridin-3-yl)methyl]piperazin-1-ium chloride. Its structure is compared to that of the salt 4--1-{[6-(4-fluorophenyl)pyridin-3-yl]methyl}piperazin-1-ium chloride monohydrate, a fluorinated analogue.The amine 8-{4-[(6-phenylpyridin-3-yl)methyl]piperazin-1-yl}-3,4-dihydroquinolin-2(1 H)-one was crystallized as the hydrochloride salt, 4--1-[(6-phenylpyridin-3-yl)methyl]piperazin-1-ium chloride, C25H27N4+·Cl− (I·HCl). The conformation of the organic cation is half-moon in shape enclosing the chloride anion. The piperidine ring of the 3,4-dihydroquinolin-2(1H)-one moiety has a screw-boat conformation, while the piperazine ring has a chair conformation. In the biaryl group, the pyridine ring is inclined to the phenyl ring by 40.17 (7) and by 36.86 (8)° to the aromatic ring of the quinoline moiety. In the crystal, the cations are linked by pairwise N—H⋯O hydrogen bonds, forming inversion dimers enclosing an R22(8) ring motif. The Cl− anion is linked to the cation by an N—H⋯Cl hydrogen bond. These units are linked by a series of C—H⋯O, C—H⋯N and C—H⋯Cl hydrogen bonds, forming layers lying parallel to the ab plane.The amine 8-{4-[(6-phenylpyridin-3-yl)methyl]piperazin-1-yl}-3,4-dihydroquinolin-2(1 The two cations differ essentially in the conformation of the biaryl group (rings B = N4/C15–C19 and C = C20–C25) and their orientation with respect to the aromatic ring (A = C4–C9) of the 3,4-dihydroquinolin-2(1H)-one moiety. This is illustrated by the view of their structural overlap, shown in Fig. 3I·HCl, pyridine ring B is inclined to phenyl ring C by 40.17 (8)° while in II·HCl the equivalent dihedral angle is 10.06 (11)°. In I·HCl, ring A is inclined to rings B and C by 36.86 (8) and 14.16 (8)°, respectively. These dihedral angles differ considerably from the dihedral angles in II·HCl, where ring A is inclined to rings B and C by 51.20 (9) and 41.40 (11)°, respectively. In both compounds, the piperidine ring (N1/C1–C4/C9) has a screw-boat conformation with the torsion angle C1—C2—C3—C4 being −56.17 (18)° in I·HCl and −55.6 (2)° in II·HCl. In both compounds, the piperazine ring (N2/N3/C10–C13) has a chair conformation.The organic cation of on Fig. 2. The molI·HCl, the organic cations are linked by a pair of N—H⋯O hydrogen bonds, forming an inversion dimer enclosing an − anion is linked to the cation by an N—H⋯Cl hydrogen bond , the cations are linked by the water molecules of crystallization via N—H⋯Ow and Ow—H⋯O hydrogen bonds to form dimers with − anions, via Ow—H⋯Cl⋯H—N hydrogen bonds, to form chains propagating along the b-axis direction. The chains are linked via C—H⋯Cl and C—H⋯O hydrogen bonds, forming layers parallel to the ab plane.In the crystal of c-axis direction and lying parallel to the ab plane. There are no significant directional inter-layer contacts present in either crystal structure.In both cases, hydrogen-bonded layers are formed stacking along the et al., 2007CrystalExplorer17  through white to blue . The Hirshfeld surfaces (HS) of I·HCl and II·HCl mapped over dnorm are given in Fig. 6a and 6b that there are important contacts present in the crystals of both compounds, the strong hydrogen bonds (Table 1Table S1) being indicated by the large red zones.The Hirshfeld surfaces are colour-mapped with the normalized contact distance, I·HCl, and those for II·HCl, are compared in Figs. 7II·HCl, amounting to 7.5%. The second most important contribution to the HS is from the C⋯H/H⋯C contacts at 20.2 and 20.5%, for I·HCl and II·HCl, respectively. These are followed by the Cl⋯H⋯Cl contacts at 10.1 and 12.8% for I·HCl and II·HCl, respectively, and O⋯H/H⋯O contacts at, respectively, 7.4 and 8.7%. The N⋯H/H⋯N contacts contribute, respectively, 6.5 and 5.3%. The C⋯C contacts in I·HCl contribute 2.9%, while the C⋯F contacts in II·HCl contribute 1.4%. All other atom⋯atom contacts contribute <1% to the HS for both compounds.The percentage contributions of inter-atomic contacts to the HS for both compounds are compared in Table 2et al., 2016H)-ones gave three hits for compounds 8-{1-[methyl]piperidin-4-yl}-3,4-dihydroquinolin-2(1H)-one (III), that crystallized as a chloroform 0.25-solvate -one (IV) -one (V)  -one (VII) -one (VIII) . Colourless plate-like crystals of their hydrochloride salts were obtained by slow evaporation of solutions in dichloromethane and methanol; ratios (8:3) and (8.5:1.5), respectively.The synthesis of compounds Uiso(H) = 1.2Ueq(C).Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989021000979/hb7965sup1.cifCrystal structure: contains datablock(s) I, Global. DOI: 10.1107/S2056989021000979/hb7965Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989021000979/hb7965sup3.pdfDetails concerning structure II.HCl. DOI: 2059142CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Sojutsu, as defined by the Japanese Pharmacopoeia, is the rhizome of Atractylodes lancea De Candolle, Atractylodes chinensis Koidzumi, or their interspecific hybrids (Asteraceae). Sojutsu is one of the traditional Kampo formulas, which are administered to patients suffering from stomach disorders, edema, and nephrotic syndrome. Although antiinflammatory effects of Sojutsu have been reported, its effects on the liver and kidney have not been extensively investigated. Here, we used a Sojutsu sample identified as A. chinensis rhizome and isolated several constituents from its ethyl acetate (EtOAc)-soluble fraction that decreased production of the proinflammatory mediator nitric oxide (NO) in interleukin 1β-treated rat hepatocytes. Among the constituents in this fraction, atractylodin showed the highest activity to suppress NO production, whereas hinesol, β-eudesmol, and α-bisabolol showed low activity. Atractylodin decreased the levels of inducible nitric oxide synthase, tumor necrosis factor α, and lipocalin 2 messenger RNAs (mRNAs). The EtOAc-soluble fraction of the A. chinensis rhizome extract was administered daily for 20 weeks to high immunoglobulin A (HIGA) mice, whose pathological findings resemble human immunoglobulin A nephropathy. This fraction decreased the weight of white adipose tissue and decreased mesangial proliferation and immunoglobulin A deposition in glomeruli. These results indicate that the EtOAc-soluble fraction, which included antiinflammatory constituents, may be responsible for improvement of the mesangial lesions in HIGA mice.The crude drug  Atractylodes species (Asteraceae) are used as medicinal herbs in East Asia, and four Atractylodes species are used as crude drugs in Japanese Kampo medicine. These species are classified by the Japanese Pharmacopoeia into two crude drugs, Sojutsu and Byakujutsu +m/z 182.0725 ; 1H NMR : δ 1.83 , 5.59 , 6.10 , 6.33 , 6.37 , 6.41 , 6.79 , 7.38 ; 13C NMR : δ 19.0 (C-9), 72.6 (C-5), 77.2 (C-4), 80.2 (C-3), 81.9 (C-6), 104.8 (C-2), 109.9 (C-7), 110.1 (C-4′), 112.1 (C-3′), 130.7 (C-1), 143.5 (C-5′), 143.7 (C-8), 151.9 (C-2′). This compound was identified as atractylodin on the basis of mass and 1H NMR and 13C NMR spectral analysis and by comparison with published data  to afford colorless needles (35.7 mg); m.p. 50–53 °C; [α]D25 +41.1° . EI–MS m/z (%): 222 , 204 , 189 (60), 161 (100); HR–EI–MS m/z 222.1966 [M]+ ; 1H NMR : δ 0.92 , 1.19 and 1.21 , 1.34–1.62 , 1.37 , 1.56 , 1.68 , 1.70 , 1.94 , 5.32 ; 13C NMR : δ 16.2 (C-12), 19.9 (C-11), 24.2 (C-8), 27.7 (C-4), 27.9 (C-3), 28.0 and 28.4 (C-14 and C-15), 33.2 (C-9), 35.7 (C-1), 36.7 (C-10), 48.4 (C-5), 51.4 (C-2), 72.0 (C-13), 121.7 (C-7), 140.1 (C-6). The assignments of these signals were based on the H–H correlation spectroscopy (COSY), heteronuclear multiple-quantum coherence (HMQC), and heteronuclear multiple-bond correlation (HMBC) spectra. The MS, 1H NMR, and 13C NMR spectral data were in agreement with reported data [α]D (+41.1° in CHCl3) of this compound was opposite to the literature value (−40° in CHCl3) of (−)-hinesol  to afford colorless needles (30.2 mg); m.p. 74.5–76 °C; [α]D21 −55.3° . EI–MS m/z (%): 222 , 204 (15), 189 (19), 164 (75), 149 (100); HR–EI–MS m/z 222.1970 [M]+ ; 1H NMR : δ 0.72 , 1.14  and 1.60  (H-6), 1.21 , 1.21 and 1.44 , 1.23–1.31 , 1.29 and 1.53 , 1.36 , 1.44 and 1.60 , 1.77 , 2.00 and 2.31 , 4.45 and 4.72 ; 13C-NMR : δ 16.3 (C-14), 22.4 (C-8), 23.5 (C-2), 25.0 (C-6), 27.1 and 27.2 (C-12 and C-13), 35.9 (C-10), 36.9 (C-3), 41.1 (C-9), 41.8 (C-1), 49.4 (C-7), 72.9 (C-11), 105.3 (C-15), 151.2 (C-4). This compound was identified as β-eudesmol on the basis of [α]D, mass, 1H NMR, and 13C NMR spectral analysis and by comparison with published data [Subfraction A5 (886 mg) was purified by silica gel column chromatography (1.5 cm i.d. × 20 cm) using hed data .y) against a series of injection amounts . The calibration equation and correlation coefficient of three standard compounds were as follows; atractylodin, y = 345,556,992x + 263,875 (R2 = 1.000); hinesol, y = 175,938,000x + 3,302,345 (R2 = 0.991); β-eudesmol, y = 197,417,365x + 6,056,334 (R2 = 0.998). Fraction A was accurately weighed and dissolved in MeOH to make a sample solution of 200 μg/mL. The sample solution (1.0 μL) was analyzed in duplicate. The peak areas of atractylodin, hinesol, and β-eudesmol in the sample solution were fit to the calibration curves, and the amounts of atractylodin, hinesol, and β-eudesmol in 1.0 μL of sample solution calculated. The amounts of atractylodin, hinesol, and β-eudesmol in 1.0 μL of sample solution (0.2 μg of fraction A) were calculated to be 0.0098 μg, 0.0490 μg, and 0.0459 μg, respectively. Therefore, the contents of atractylodin, hinesol, and β-eudesmol in fraction A were 4.88%, 24.5%, and 23.0%, respectively. α-Bisabolol was not quantified because it was present in trace amount, but its presence was confirmed by GC–MS. The fragment pattern of the peak at tR 17.9 min was consistent with that of α-bisabolol (data not shown).Atractylodin, hinesol, β-eudesmol, and α-bisabolol were analyzed by GC–MS using a DB-5MS capillary column . The analytical conditions were as follows: ionization mode EI, injector and transfer line temperature, 200 °C; oven temperature programmed from 50 to 300 °C at 10 °C/min; carrier gas, helium (1 mL/min); splitless mode; ionization voltage, 70 eV; ionization current, 300 μA. Commercially purchased (−)-α-bisabolol was used as standard. Atractylodin, hinesol, β-eudesmol, and α-bisabolol were accurately weighed and dissolved in MeOH to make stock solutions of 1.0 mg/mL. Then, series of hinesol and β-eudesmol standard solutions  were prepared by diluting the stock solutions to make calibration curves. Series of atractylodin and α-bisabolol standard solutions  were similarly prepared. A calibration curve of each standard compound was calculated by plotting peak areas , male Wistar rats  were housed at 21–23 °C under a 12 h light–dark cycle and fed with a CRF-1 diet  and had water available ad libitum. Before preparation of hepatocytes, the animals were acclimated to their housing for a week.n = 3–4 per group). At age of 20 and 30 weeks, three mice in each group were euthanized by cervical dislocation, and blood was drawn from the heart to obtain serum. Perirenal and parametrial white adipose tissue was excised and weighed. The kidneys were excised and soaked in RNAlater stabilization solution  for subsequent RNA preparation and histochemistry.Female HIGA mice and female BALB/c mice  were fed daily with a CRF-1 diet with water available ad libitum. BALB/c mice were used as healthy controls. These mice were fed with a CRF-1 diet with or without 1% (w/w) fraction A of ACR extract from 10 weeks until 30 weeks of age , according to a previously published method , 25. Bri50) value against nitrite was determined in triplicate for at least three different concentrations of an extract or constituent [50 value of an extract or constituent was calculated to determine its ability to suppress NO production, unless the compound was cytotoxic. The LDH activity in the medium was measured using an LDH cytotoxicity detection kit  to estimate cytotoxicity.Each fraction or constituent was added to the medium on day 1, and the hepatocytes were incubated for 8 h. Nitrite (a stable metabolite of NO) in the medium was measured using the Griess method to measure NO levels , 27. The2 and incubated at 37 °C for 1.5 h [2 alone was set at 100%.Each constituent was added to a medium containing 25 μM NaNOor 1.5 h . This meWestern blotting was performed, as previously described . Brieflylater Solution) using Sepasol I Super G solution  and purified with the RNAqueous kit and the TURBO DNA-free kit (Applied Biosystems) [EF mRNA [Total RNA was prepared from rat hepatocytes and the kidney of each mouse (in RNAsystems) , 16. TheSerum was obtained from blood drawn from mouse hearts, and the levels of blood urea nitrogen (BUN) and creatinine were measured. The serum levels of LCN2 and G-CSF were determined by enzyme-linked immunosorbent assay (ELISA) using a Mouse NGAL ELISA kit  and a Mouse G-CSF Quantikine ELISA kit .later solution) were fixed in 4% formaldehyde and embedded in paraffin. Sections (3–5 mm) were deparaffinized and stained with hematoxylin and eosin. To detect IgA deposition in the glomeruli, the deparaffinized sections were treated with 3% hydrogen peroxide, blocked with 10% normal goat serum , and incubated with a goat anti-mouse IgA antibody conjugated with HRP  at 23 °C for 2 h. Immunoreactivity was detected with 3,3′-diaminobenzidine tetrachloride (Nichirei Bioscience) as substrate. The resultant sections were counterstained with hematoxylin and examined under a BA210EINT research biological digital microscope . The glomeruli showing immunoreactivity to IgA were quantified in a blinded manner, and 10 low-power fields  were counted five times.Kidney specimens taken from 30-week-old mice . The differences were analyzed using Student’s Atractylodes species present in the crude Sojutsu drug used in this study was unknown; therefore, the genomic DNA of the Sojutsu was subjected to PCR and subsequent DNA analyses. As references, we used leaves of A. lancea, A. chinensis, and A. japonica that were botanically authenticated. The ITS1 sequence between the 18S–5.8S rRNA genes [Sojutsu samples was compared with those obtained from the leaves. The 167-base-pair (bp) ITS1 sequence from five pieces of the Sojutsu sample was identical to those of A. chinensis leaf, i.e., 100% homology, whereas it was 99.4% (166 bp/167 bp) and 97.0% (162 bp/167 bp) identical to those of the A. lancea and A. japonica leaves, respectively. These data indicate that the Sojutsu sample used in this study was an A. chinensis rhizome. To avoid confusion, we hereafter call this Sojutsu sample “Atractylodes chinensis rhizome,” instead of “Atractylodes lancea rhizome,” as defined by the Japanese Pharmacopoeia [The NA genes , 3 derivEF gene of the Sojutsu sample was also examined by sequencing because EF is a ubiquitously expressed housekeeping gene. The 230-bp EF sequence of the Sojutsu sample was identical to those of the A. lancea and A. japonica leaf samples. These data indicate that the EF gene sequences are highly conserved among Atractylodes species and cannot be used to discriminate Atractylodes species.The n-butanol (fraction B), and water (fraction C) . When the NO-quenching activity of atractylodin was measured, no significant changes in NO levels were observed compared with the NO levels in the medium containing NaNO2 alone (data not shown), indicating that atractylodin did not directly quench NO. Therefore, we used atractylodin for the subsequent experiments.Next, we examined whether these constituents in ACR fraction A could suppress NO induction in IL-1β-treated hepatocytes Table . Among tiNOS gene expression were further examined. When atractylodin was added with IL-1β to the hepatocyte culture medium, it inhibited the induction of NO and the iNOS protein , chemokine C–C motif ligand 2 (CCL2), and lipocalin 2 (LCN2) mRNAs in IL-1β-treated hepatocytes  mice. IgA-positive glomeruli were 82.0% of the total number of glomeruli. In contrast, weaker immunoreactivity to IgA was detected in the glomerular mesangial matrix of HIGA + ACRA mice, and the number of IgA-positive glomeruli was 57.0%. As shown in Fig. Kidney function was assessed by analyzing serum samples of 20- and 30-week-old mice  mice tended to be higher than those in BALB/c(−) and HIGA + ACRA mice, these differences were not significant . ELISA of mouse serum indicated that LCN2 levels in the HIGA + ACRA group were slightly lower than those of the HIGA(−) group, but these differences were not significant  is produced from monocytes and macrophages during inflammation , and theSojutsu sample defined by the Japanese Pharmacopoeia [A. chinensis by genomic DNA sequencing of rRNA genes. When an EtOAc-soluble fraction (fraction A) was prepared from an extract of this ACR, it significantly decreased iNOS gene expression in primary cultured rat hepatocytes, in which the iNOS gene has been induced by IL-1β [iNOS gene expression in IL-1β-treated hepatocytes  to produce NO and proinflammatory cytokines, whereas rat hepatocytes do not respond to LPS . Therefoot shown , 11. Accot shown , it is piNOS mRNA levels  mice Table . YoshimuSaireito, which includes Sojutsu, shows antiinflammatory effects in rat hepatocytes [Saireito  was administrated daily to HIGA mice, the pathological findings in the glomeruli of the kidney were not significantly improved (data not shown). It is possible that the dose of Saireito (1% powder in the diet) and the dose of Sojutsu constituents in Saireito were too low.The Kampo formula atocytes . When Saklotho-deficient mice, which is a model of human aging [A. chinensis and A. lancea, a future study on the A. lancea rhizome is also warranted. These studies will lead to effective use of Sojutsu for kidney disease.Although atractylodin in ACR fraction A may be involved in the improvement of HIGA mouse pathology, this remains to be confirmed. Recently, Fujitsuka et al. reported that administration of atractylodin increased ghrelin signaling and prolonged survival in an aging . A varieSojutsu, the extract from the ACR was examined for its antiinflammatory properties. Using IL-1β-treated hepatocytes, atractylodin and three sesquiterpenes were identified as constituents in an EtOAc-soluble fraction of an ACR extract (fraction A). Because atractylodin effectively suppressed NO production and iNOS gene expression, it may be primarily responsible for the antiinflammatory activity of the ACR extract. Daily administration of the EtOAc-soluble fraction for 20 weeks improved the mesangial lesions in the renal glomeruli of IgA nephropathy model mice. Although further investigation is necessary to confirm the involvement of atractylodin in this improvement, this study provides a basis for developing the use of Sojutsu (A. chinensis and A. lancea) and atractylodin in the treatment of human kidney disease.As an example of


The title compound is a Schiff base formed from 5-nitrothiophene-2-carbaldehyde and 2-methyl-3-nitroaniline. In the crystal, the molecules are linked by weak C—H⋯O hydrogen bonds. 12H9N3O4S, synthesized by condensation of 5-nitrothiophene-2-carbaldehyde and 2-methyl-3-nitroaniline, crystallizes in the orthorhombic space group P212121. In the molecule, the aromatic benzene and thiophene rings are twisted with respect to each other, making a dihedral angle of 23.16 (7)°. In the crystal, molecules are linked by intermolecular C—H⋯O hydrogen bonds into chains extending along the c-axis direction. Weak π–π stacking interactions along the a-axis direction provide additional stabilization of the crystal structure. The roles of the various intermolecular interactions were clarified by Hirshfeld surface analysis, which reveals that the crystal packing is dominated by O⋯H (39%) and H⋯H (21.3%) contacts. The crystal studied was refined as a two-component inversion twin.The title compound, C In this respect, Schiff bases are organic molecules suitable for bioactivity applications because of the imine bond that increases the lipophilic character of the molecule. The imine bond provides a synthetic route to structural chirality, changes the electronic properties and leads to solubility in different media °. The deviation from planarity can be attributed to packing forces. The nitro group attached to the thiophene ring is strongly conjugated with the π-system of this ring, as evident from the short N2—C7 distance (see Table 1N-[(Z)-4-propoxybenzylidene]-1,3-thiazol-2-amine phenol -4,5-dihydropyridazin-3(2H)-one aniline gave 15 hits including 4-methyl-N-[(5-nitrothiophen-2-yl)methylidene]aniline -1-(5-nitrothiophen-2-yl)methanimine methanimine  shows donor (red) and acceptor (blue) regions. The percentage contribution of various interactions is shown in the fingerprint plot  and 2-methyl-3-nitroaniline  in ethanol (40 ml) for 5 h under stirring. The obtained yellow crystalline material was washed with ethanol and dried at room temperature . Crystals were grown from a solution in ethanol.Uiso(H) = 1.5Ueq(C-methyl) and 1.2Ueq(C) for other C-bound H atoms. The structure was refined as a two-component inversion twin.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989021000529/yk2142sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021000529/yk2142Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021000529/yk2142Isup3.cmlSupporting information file. DOI: 2055920CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


B. radicata fermentation broth, the broth was purified by DEAE-cellulose and sephadex LC-20 column. The compounds were submitted to spectral analyses .To evaluate the antimicrobial and microbicidel activity of 37O10H43N and 661 Da. only Griseococcin (1) has good antimicrobial activity among the Griseococcin(s). The zone of inhibition (ZOI), minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) or minimum fungicidal concentration (MFC) of Griseococcin (1) were used to investigate the antimicrobial activity. Antifungal activity of Griseococcin (1) was significant, especially for main pathogenic fungus Trichophyton rubrum and Trichophyton mentagrophytes, MFC/MIC of Griseococcin (1) was 1, while MFC/MIC of postive control was greater than 4, the fungicidal effect of Griseococcin (1) was better than that of positive control.The purified compounds were identified as the Griseococcin(s) which were naphthoquinone derivatives, the Chemical formula and MW of Griseococcin (1) was determined as CB. radicata was purified. The purified compound can restrain main pathogens (T. rubrum and T. mentagrophytes) leading to tinea pedis. The antifungal activity of Griseococcin (1) was similar to that of the positive control and the fungicidal effect of Griseococcin (1) was better than that of positive control, it might be suitable for pharmaceutical industries.In this paper, the secondary metabolite compound Griseococcin (1) from  Trichophyton rubrum and Trichophyton mentagrophytes , four hydroxyl groups at δ H 8.37 , δ H 7.81  and δ H 7.80  and 9.63 .Griseococcin 1) was isolated as a white amorphous solid powder with the molecular formula of C was isol1H-1HCOSY, HMBC, and HSQC spectra  to C-8 (δC 135.45) and C-6 (δC 137.21), from H3–14′ (δH 1.85) to C-2′ (δC 215.7) and C-4′ (OH) (δC 73.60), from H3–15′ (δH 2.11) to C-6′ (δC 215.70) and C-4′ (OH) (δC 73.60), from H2–7′ (δH 1.08) to C-9′ (δC 71.25) and C-13′ (δC 71.18). The 1H, 1H three-bond couplings observed in the COSY spectrum from H-8′ (δH 1.94) to H-9′ (δH 3.62), from H-10′ (δH 1.29) to H-11′ (δH 3.49), from H-12′ (δH 1.73) to H-13′ (δH 3.51), together with the chemical shifts of the 13C resonances (C-8′-13′) observed at alternating higher and lower fields, revealed the presence of cyclohexane with alternating hydroxyl and methyl groups. 1H-1H COSY correlations from H2–13  to H2–14 , from H2–14  to H2–15  and from H2–16  to H2–17  and HMBC correlations from H2–13  to C-15 (δC 166.02), from H2–14  to C-16 (δC 166), from H2–15  to C-17 (δC 166.01) and from H2–16  to C-18 (δC 23.15) identified coupled olefins. The key HMBC correlations from H2–1′  to C-3′ (δC 23.4), from H − 3′  to C-5′ (δC 29.05), from H3–14′  to C-2′ (δC 215.7) and C-4′-OH (δC73.6), from H3–15′  to C-6′ (δC 215.7) and C-4′-OH identified two meta position carbonyl group and one ortho position hydroxyl group  was deduced by comprehensive analysis of tra Fig. a. In Gri3–14′ to C-3′ and from H3–15′ to C-6′. Therefore, the complete structure of naphthoquinone was determined as shown in Fig. This connectivity was also secured by the observation of the HSQC correlations from HGriseococcin (1) was white powder and its solubility was 0.063 g/ml in water. It could be slight soluble in methanol and DMSO, but insoluble in n-hexane, dichloromethane, chloroform, ethyl acetate and acetone.− 1 represented the stretching vibration of O–H. The weak absorption peaks at 2356 and 2925.5 cm− 1 were resulted from the stretching vibration of C–H. The absorption bands at 1637.4 and 1618.1 cm− 1 are due to the vibration of C=C and C=O in the ester group. The absorptions peaks at 1456.1, 1414 and 624 cm− 1 were attributed to the presence of an internal C–H deformation. The strong absorption peak at 866 cm− 1 was resulted from aromatics. In conclusion, the typical absorption peak indicated that Griseococcin (1) was naphthoquinone with group O–H,C-H,C=C,C=O and so on [The FT-IR spectrum of Griseococcin (1) showed Fig.  an intenTrichophyton rubrum (ATCC 28188), Trichophyton mentagrophytes (ATCC 9533), Epidermophyton floccosum (ATCC 52066), Candida albicans (ATCC 10231), Staphylococcus aureus (ATCC 6538), Bacillus subtilis (ATCC 6051), Escherichia coli (ATCC 8739) and Pseudomonas aeruginosa (ATCC 27582) The results were shown in Table T. rubrum, T. mentagrophytes with ZOI values of 18.06 ± 0.85, 15.01 ± 1.02 mm, as compared to the positive control with ZOI = 20.67 ± 1.58, 28.33 ± 2.15 mm, respectively. While antibacterial activity was weak.Griseococcin 1) was assessed for antimicrobial and microbicidel activity against selected  was asseT. rubrum), the MIC, MFC and MFC/MIC values were 31.2 ± 2.7, 31.2 ± 3.1 μg/ml and 1, while MIC, MFC and MFC/MIC values of Terbinafine were 15.6 ± 1.2, 31.2 ± 1.6 μg/ml and 6. Fungicidal activities of Griseococcin (1) are revealed more effective than that of commercial reagents (Terbinafine).J.Meletiadis et al. reported that compounds were considered bactericidal or fungicidal when the MBC/MIC or MFC/MIC ratio is ≤4 . In thisS. aureus, E. coli and P. aeruginosa). The results were shown in Table S. aureus and E. coli,. MBC/MIC value of Griseococcin (1) was 2.0, while the MBC/MIC ratio was 3.0 and 4.0 for positive control (Gentamicin sulfate).Griseococcin (1) also showed high bactericidal activities with MIC, MBC and MBC/MIC values ranged between 62.5 ~ 125 μg/ml, 125–500 μg/ml and 2–4 against examined bacteria  Pat) had remarkable antifungal activities. These data are consistent with previous findings on the minimum inhibitory concentrations (MICs) and zone of inhibition (ZOI) of B. radicata fermentation [In the present study, Griseococcin (1) purified from selected puffball  on fungi showed that the most sensitive fungi strains were the main pathogenic fungi (T. rubrum and T. mentagrophytes) causing tinea pedis, ZOIs were 18.06 ± 0.85 and 15.01 ± 1.02 mm, MICs were 31.2 ± 2.7 and 31.2 ± 1.8 μg/ml, MFCs were 31.2 ± 3.1 μg/ml and 31.2 ± 2.1 μg/ml, MFC/MICs were 1 and 1 against T. rubrum and T. mentagrophytes. ZOI values of positive control (Terbinafine) were 20.67 ± 1.58 mm 28.33 ± 2.15 mm, MICs were 15.6 ± 1.6 and 7.8 ± 1.2 μg/ml, MFCs were 93.6 ± 2.1 and 39.0 ± 2.2 μg/ml, MFC/MICs were 6 and 5 respectively. The antifungal effect of Griseococcin (1) was similar with that of positive control, the fungicidal effect of Griseococcin (1) was better than that of positive control. The most sensitive bacterial species for Griseococcin (1) was S. aureus and E. coli, MICs and MBCs were 62.5 ± 1.5, 125 ± 2.3 and 125 ± 3.1, 250 ± 2.1 μg/ml respectively, P. aeruginosa and B. subtilis were more resistant. MFC/MICs and MBC/MICs of Griseococcin (1) were less than positive control which meant that antibacterial activity Griseococcin (1) was better than that of the commercial drugs. This study is important for the development of new drugs with low toxicity, overcoming drug resistance and recurrence.In this study, the purification extraction Griseococcin(s) from fermentation broth of 37H43NO10 and the molecular structure of Griseococcin (1) was shown in Fig. The FT-IR spectrum of Griseococcin 1) showed the strong absorption band, stretching vibration and bending vibration of O-H, C=O, C=C and C-H which belong to a unsaturated coupled bond and aromatic form of naphthoquinone. According to HR-ESI-MS analysis, MW of Griseococcin (1) was 661 Da. Based on the results of different spectral  studies and physicochemical properties, the molecular formula of Griseococcin (1) was C showed tB. radicata was another important biological function. The biological activities of organic compounds are related to their molecular weight, functional groups, the length of chain, the composition of group and the number of branches, hydrophilic and hydrophobic group. It means that the structure-activity relationship should be disclosed.Previously, many authors reported the various biological activity of fermentation broth from puffball like anticancer activity , 27, antFuture work concentrating on determining the antifungal mechanisms of Griseococcin (1) will be performed, which will be helpful in laying a foundation for overcoming the drug resistance that pathogens quickly develop against tinea pedis.B. radicata were studied. The compound from Bovistella radicata (Mont.) Pat was purified. Molecular weight and molecular formula of the purified compound (Griseococcin (1)) were 661 Da and C37H43NO10 respectively, it can restrain main pathogens (T. rubrum and T. mentagrophytes) leading to tinea pedis. The antifungal activity of Griseococcin (1) was similar to that of the positive control.In this paper, the antifungal secondary metabolite compound Griseococcin (1) from Bovistella radicata (Mont.) Pat was obtained from Shuinan town, Jishui county, Jiangxi province, China. After strain identification and authenticated by Professor Qingmei Zeng, it belongs to Agaricalesorder, Lycoperdaceae family, Bovistella genus, Bovistella radicata (Mont.) Pat species. The tested pathogenic fungi included Trichophyton rubrum (ATCC 28188), Trichophyton mentagrophytes (ATCC 9533), Epidermophyton floccosum (ATCC 52066), and Candida albicans (ATCC 10231). Four strains of test pathogenic bacteria included Staphylococcus aureus (ATCC 6538), Bacillus subtilis (ATCC 6051), Escherichia coli (ATCC 8739) and Pseudomonas aeruginosa (ATCC 27582). All standard bacterial and fungal strains were obtained from rom the Microbiology Laboratory at Department of Biology, Anhui Medical University, Anhui.B.radicata should be dried at 40 °C for at least 2 days, and taken out when its weight is no longer changed, the sporophore and spore powder were ground together and filtered through a 100 mesh sieve. The mixed powder of B.radicata was inoculated into 100 mL of potato dextrose broth (PDB) in 250 ml flask. The flask was kept in rotary shaker at 25 °C with 115 rpm for 72 h. The pH and moisture content of PDB was also determined according Maguireboyle (2014) and Mcauliffe (2016) [T. rubrum and T. mentagrophytes by zone of inhibition (ZOI) method. Then the fermentation were centrifuged at 7000 rpm for 20 min and filtered over Whatman No.4 paper to get the final clear supernatant and preserve at 4 °C. 50 ml clear supernatant was purified firstly using 100 ml DEAE-cellulose column and eluted by different concentration NaCl (10–30%) to get different fractions. 20% NaCl elution fraction showed best antifungal activity against pathogens and was named as SPAF. Furthermore, SPAF (20% NaCl elution fraction) was purified by sephadex LH-20 column. Different purified fractions (named Griseococcin(s)) were obtained from SPAF, only Griseococcin(1) (500 μg/ml) has antifungal activity and it’s biochemical characteristics and spectral  studies were assessed.The mature e (2016) , 31. For6 CFU/mL) of T. rubrum, T. mentagrophytes, E. floccosum, and C. albicans in potato dextrose broth and S. aureus, B. subtilis, E. coli and P. aeruginosa in Nutrient Broth [S. aureus, E. coli, B. subtilis and P. aeruginosa, for 48 h with T. rubrum, T. mentagrophytes, E. floccosum, and C. albicans. Griseococcin (1) was also dissolved in NS at 1 mg/mL. The MIC was recorded as the lowest concentration of sample showing no detectable growth. MFC or MBC was determined as the concentration causing no visible growth and killing 99.5% of the original inoculum. Ten microliters of sub-inhibitory concentrations of Griseococcin (1) was placed in the corresponding solid medium for 48 h to determine the MBC or MFC values according to the growth of the microbial colonies. Each sample was performed twice. The zones of inhibition (ZOI) of Griseococcin (1) (100 μg/ml) was also evaluated. The prepared Griseococcin (1) was filled into the wells. After incubating for 24 h at 37 °C, the measurements were done basically from the edge of the zone to the edge of the well [The examined methods were the minimum inhibitory concentrations (MICs) , minimumnt Broth  were inothe well .max absorption spectrum of SPAF was analyzed at full-wave spectra (200–900 nm) by UV/vis 2802 spectrophotometer. The FT-IR spectrum of Griseococcin(s) were recorded on a Thermo Nicolet Spectrum FT-IR in a range of 4000–400 cm− 1 with KBr pellets. HR-ESI-MS data were obtained on an Agilent 1260 Infinity LC coupled to a 6230 TOF. 20 mg of the dried sample was dissolved in 0.55 mL of deuteroxide (99.99% D) in a NMR tube. 1D and 2D NMR spectra were acquired on an AVANCE-600 NMR spectrometer  at 50 °C. The chemical shifts were given in δ (ppm) and referenced to the solvent signal . Column chromatography (CC) was conducted on DEAE-cellulose and Sephadex LH-20. The fractions Griseococcin(s) were also monitored by HPLC  which was equipped with a diode array detector (DAD). The DAD detector was set at 215 nm to acquire chromatograms. The separation of the compound was performed on a Hypersil RP-C18 column  at a temperature of 25 °C. Injection volume: 20 μL.The UV− 1; UV (D2O) λ max 215 nm; 1H and 13CNMR data see Table 37 H43NO10, 661.1968).Griseococcin (1): IR (neat) v max 3417, 2926, 2356, 1637, 1618, 1456, 1414, 866, 624 cmAdditional file 1.


N,N-dimethylmethanamine) and N,N′,N′′,N′′′-tetrakis(N-methylaniline), and a degredation product, N,N′-[bis(methylene)]bis(N-methylaniline), are described and have been analysed by Hirshfeld surface analysis.The crystal structures of two new tetrakis-substituted pyrazine compounds, 1,1′,1′′,1′′′-tetrakis, C16H32N6, (I) and N,N′,N′′,N′′′-tetrakis(N-methylaniline), C36H40N6, (II), both crystallize with half a molecule in the asymmetric unit; the whole molecules are generated by inversion symmetry. There are weak intramolecular C—H⋯N hydrogen bonds present in both molecules and in (II) the pendant N-methylaniline rings are linked by a C—H⋯π interaction. The degredation product, N,N′-[bis(methylene)]bis(N-methylaniline), C28H29N5, (III), was obtained several times by reacting (II) with different metal salts. Here, the 6-phenyl ring is almost coplanar with the planar pyrrolopyrazine unit (r.m.s. deviation = 0.029 Å), with a dihedral angle of 4.41 (10)° between them. The two N-methylaniline rings are inclined to the planar pyrrolopyrazine unit by 88.26 (10) and 89.71 (10)°, and to each other by 72.56 (13)°. There are also weak intramolecular C—H⋯N hydrogen bonds present involving the pyrazine ring and the two N-methylaniline groups. In the crystal of (I), there are no significant intermolecular contacts present, while in (II) molecules are linked by a pair of C—H⋯π interactions, forming chains along the c-axis direction. In the crystal of (III), molecules are linked by two pairs of C—H⋯π interactions, forming inversion dimers, which in turn are linked by offset π–π interactions [intercentroid distance = 3.8492 (19) Å], forming ribbons along the b-axis direction.The two new tetrakis-substituted pyrazines, 1,1′,1′′,1′′′- tetrakis( A search of the Cambridge Structural Database  tetrakis (I)N,N′,N′′,N′′′-tetrakis(N-methylaniline) (II)N,N′-[bis(methylene)]bis(N-methylaniline) (III)In our search for new tetrakis-substituted pyrazine ligands  −x, −y, −z 1 ], linking the two dimethylmethanamine substituents ts Fig. 1.The molecular structure of compound (II)b]pyrazine unit , forming a dihedral angle of 4.41 (10)°. On the other side of the molecule, the two adjacent N-methylaniline rings (C14–C19 and C22–C27) are inclined to the planar pyrrolopyrazine unit by 88.26 (10) and 89.71 (10)°, and to each other by 72.56 (13)°. There are also weak intramolecular C—H⋯N hydrogen bonds present involving the pyrazine ring and the two N-methylaniline groups ps Fig. 3.In the crystal of (I)In the crystal of (II)g3⋯Cg6ii, where Cg3 and Cg6 are, respectively, the centroids of the phenyl ring (C7–C12) and the pyrrolopyrazine ring system, has a centroid–centroid distance of 3.8492 (14) Å, α = 4.41 (10)°, interplanar distances of 3.6495 (14) and 3.5490 (7) Å, with an offset of 1.49 Å .In the crystal of (III)et al., 2007CrystalExplorer17  through white to blue . The Hirshfeld surfaces (HS) of the title compounds, mapped over dnorm, are given in Fig. 7a and 7b that there are no contact distances shorter than the sum of the van der Waals radii in the crystals of either compounds (I)c), two small red spots indicate the presence of weak C⋯H contacts , 68.6% for (II)b), and 63.3% for (III)c). The second most important contribution to the HS for compound (I)The two-dimensional fingerprint plots for the title compounds are given in Fig. 8et al., 2016catena-tricobalt(II)] trans-diaquadiiron(II)] dihydrate  polymer chain, catena-manganese dichloride dihydrate] pyrazine]bis(μ2-chloro)dichlorodicopper hydrate] pyrazine yielded only eight hits, of which five compounds are polymeric; see for example et al., 2017viz. catena-[bisdizinc unknown solvate] bis(μ-hydroxo)bistrinickel unknown solvate] , has been shown to be extremely successful in forming 17 metal–organic frameworks (MOFs). It was designed by Jiang and coworkers pyrazine Synthesis of 1,1′,1′′,1′′′- tetrakis (I)2NH formed in situ as a gas and was directly condensed in a round-bottom flask in an acetone/liquid N2 bath at about 213 K using a weak vacuum. Once a sufficient quantity of liquid amine had formed, a solution of 2,3,5,6-tetrakis(bromomethyl)pyrazine  in 50 ml of CH2Cl2 was added dropwise at low temperature (ca 243 K). The reaction was left for about 4 h, allowing the temperature rise to RT. The excess amine was allowed to evaporate off before the solvent was gassed off. The residue obtained was dissolved in 40 ml of MeOH and passed through a resin column (15 g of Dowex 1 X8) previously charged with OH− ions in order to exchange the HBr molecules, still attached to the ligand, by H2O molecules. About 150 ml were used as eluent. Solvent evaporation yielded 0.27 g (87%) of a light-yellow powder of compound (I)A large excess of dimethyl amine hydrochloride in water was neutralized with NaOH in an ice bath. Me1H NMR : 3.65 , 2.15 . 13C NMR : 152.16, 62.53, 46.54. IR : 2974 (s), 2942 (s), 2854 (m), 2820 (vs), 2772 (vs), 1635 (b), 1456 (s), 1414 (m), 1348 (s), 1259 (s), 1204 (m), 1168 (m), 1027 (vs), 987 (m), 841 (s). MS , m/z: 310 (MH+), 264, 178. Anal. for C16H32N6 (308.5 g mol−1) Calculated (%) C 62.30, H 10.46, N 27.24. Found (%) C 61.86, H 10.73, N 27.50.Synthesis ofN,N′,N′′,N′′′-tetrakis(N-methylaniline) (II):3CN was added dropwise to a suspension of N-methylaniline  and Na2CO3  in 25 ml of CH3CN. The colour changed immediately from light to deep yellow. The mixture was refluxed for ca 2 h, followed by TLC and then cooled to RT. The white precipitate (NaBr and excess Na2CO3) was filtered off and the filtrate was evaporated under vacuum. The residue was dissolved in hexane and the insoluble yellow powder obtained was recovered, washed with more hexane and then dried to yield 0.335 g (60%) of compound (II)3 solution of (II)A solution of 2,3,5,6-tetrakis(bromomethyl)pyrazine  in 35 ml of CH1H NMR : 7.14 , 6.68 , 4.58 , 2.79 . 13C NMR : 149.64, 149.31, 128.94, 116.92, 113.17, 54.75, 39.46. IR : 2926 (w), 1601 (s), 1508 (vs), 1446 (m), 1377 (m), 1366 (m), 1313 (m), 1257 (m), 1212 (m), 1117 (w), 993 (w), 820 (w), 745 (s), 689 (m). MS , m/z: 594 (MK+), 374, 291. Analysis for C36H40N6 (556.7 g mol−1) Calculated (%) C 77.66, H 7.24, N 15.09. Found (%) C 76.82, H 7.19, N 15.07.Synthesis ofN,N′-[bis(methylene)]bis(N-methylaniline) (III)4)2 (in excess), MnCl2·4H2O and Ni(AcO)2·4H2O. No spectroscopic or other analytical data are available for this compound.Hexagonal pale-yellow plate-like crystals of compound (III)Uiso(H) =1.5Ueq(C-methyl) and 1.2Ueq(C) for other H atoms. Note for compound (III)Crystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989020002133/xi2023sup1.cifCrystal structure: contains datablock(s) I, II, III, Global. DOI: 10.1107/S2056989020002133/xi2023Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989020002133/xi2023IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989020002133/xi2023IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989020002133/xi2023IIsup5.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989020002133/xi2023IIIsup6.cmlSupporting information file. DOI: 1984024, 1984023, 1984022CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information: