Mutations can occur in Gα proteins which prevent GTP hydrolysis; this allows the G proteins to signal independently of G protein–coupled receptors and can result in various cancers, such as uveal melanoma (UM). Most UM cases harbor Q209L, Q209P, or R183C mutations in Gαq/11 proteins, rendering the proteins constitutively active (CA). Although it is generally thought that active, GTP-bound Gα subunits are dissociated from and signal independently of Gβγ, accumulating evidence indicates that some CA Gα mutants, such as Gαq/11, retain binding to Gβγ, and this interaction is necessary for signaling. Here, we demonstrate that disrupting the interaction between Gβγ and Gαq is sufficient to inhibit aberrant signaling driven by CA Gαq. Introduction of the I25A point mutation in the N-terminal α helical domain of CA Gαq to inhibit Gβγ binding, overexpression of the G protein Gαo to sequester Gβγ, and siRNA depletion of Gβ subunits inhibited or abolished CA Gαq signaling to the MAPK and YAP pathways. Moreover, in HEK 293 cells and in UM cell lines, we show that Gαq-Q209P and Gαq-R183C are more sensitive to the loss of Gβγ interaction than Gαq-Q209L. Our study challenges the idea that CA Gαq/11 signals independently of Gβγ and demonstrates differential sensitivity between the Gαq-Q209L, Gαq-Q209P, and Gαq-R183C mutants.Heterotrimeric G protein stimulation  Q209L or Q209P mutations in Gαq or Gα11 occur in 90% of UM patients, and mutations at R183 occur in about 5% of patients (Heterotrimeric G proteins (Gαβγ) are canonically activated through G protein–coupled receptors (GPCRs), which allows for GDP release from the Gα subunit and further GTP binding. It is generally accepted that GTP binding on the Gα subunit promotes an active conformation and allows for the dissociation from Gβγ. Active GTP-bound Gα proteins can further bind and stimulate downstream effectors , 2. Hydrc tumors , 6. Mutapatients . Approxipatients .q/11 in UM, driven by the activating Q209L/P and R183 mutations, promotes the stimulation of two major signaling pathways: 1) the mitogen-activated protein kinase (MAPK) pathway and 2) the Yes-associated protein and Transcriptional coactivator with PDZ-binding motif (YAP/TAZ) pathway. The MAPK cascade is stimulated by the direct binding of CA Gαq/11 to phospholipase C–β (PLC-β), which hydrolyzes phosphatidylinositol 4,5-bisphosphate into diacylglycerol and inositol 1,4,5-trisphosphate. Diacylglycerol further activates PKC, which phosphorylates and activates RasGRP3. The further activation of Ras promotes the stimulation of the MAPK cascade and results in the phosphorylation and activation of ERK. Phospho-ERK (pERK) dimerizes and translocates into the nucleus to bind transcription factors and promote the transcription of proliferative genes  transcription factor in the nucleus to promote the transcription of genes involved in proliferation and cell survival  and FR900359 (FR) have been shown to inhibit both WT and CA Gαq/11 and have shown promising results in cancer cell models , Q209P (QP), and R183C (RC)—is sufficient to inhibit oncogenic signaling of these mutationally active Gαq. We provide evidence that disrupting the interaction between CA Gαq and Gβγ significantly inhibits Gαq-mediated activation of the MAPK and YAP/TAZ pathways. Surprisingly, we show differential sensitivity between Gαq-QL and the Gαq-QP/RC mutants in that oncogenic signaling by Gαq-QP and Gαq-RC is more sensitive to the disruption of Gβγ binding than Gαq-QL.Here, we tested the hypothesis that disrupting the interaction between Gβγ and the CA Gαq, such as PLC-β, requires the binding of Gαq to Gβγ , Q209P (QP), and R183C (RC) mutants. To monitor the activity of the YAP/TAZ pathway, we transfected the CA mutants, with and without the I25A mutation, into HEK 293 Gαq/11 CRISPR-Cas9 KO cells and monitored YAP activity using the TEAD luciferase reporter assay, which is stimulated in response to YAP activation and nuclear translocation. Gαq-QL, Gαq-QP, and Gαq-RC all showed strong activation in the assay. Cells expressing the pcDNA3 vector and WT Gαq were used as negative controls and indicated no significant change in TEAD luciferase reporter activity (A). The Gαq-QL-C9,10S palmitoylation-deficient mutant was used as a negative control for Gαq-QL. The C9,10S mutation in WT Gαq has been previously established to prevent plasma membrane localization and consequently prevent GPCR-dependent activation . It has been previously established that activation of the Rho-dependent YAP pathway results in stabilization and dephosphorylation of YAP and its subsequent translocation into the nucleus  as a control resulted in the loss of nuclear YAP and its accumulation in the cytoplasm. Gαq-I25A-QL also promoted nuclear localization of YAP, consistent with its retention of signaling in the TEAD-luciferase assay (A). Cells expressing CA Gαq-QP and Gαq-RC had robust nuclear YAP localization, indicating stimulation of this pathway. However, Gαq-I25A-QP and Gαq-I25A-RC failed to stimulate the nuclear localization of YAP, as indicated by the cytoplasmic localization of YAP. Similar to the TEAD-luciferase reporter assay results, this data suggests that Gαq-I25A-QL is more resistant to disruption of signaling than the Gαq-I25A-QP and Gαq-I25A-RC mutants. In immunofluorescence microscopy studies, we also examined the subcellular localization of the CA Gαq mutants with and without the I25A mutation . Expression of CA Gαq-QL resulted in pERK stimulation >4-fold compared to cells transfected with pcDNA3 or WT Gαq. The Gαq-I25A-QL mutant also activated pERK at a slightly decreased level compared to Gαq-QL, although the difference was not statistically significant. Interestingly, the Gαq-QP and Gαq-RC mutants stimulated pERK, but pERK activity was significantly decreased in cells expressing the Gαq-I25A-QP and Gαq-I25A-RC mutants (C and D). The effects of the I25A mutants on MAPK activity were further validated with the serum response element (SRE) luciferase reporter assay, which monitors both Rho and MAPK-dependent activity (E). As indicated, the CA Gαq-QL mutant robustly stimulated SRE-dependent luciferase activity. Introduction of the I25A mutation into Gαq-QL (Gαq-I25A-QL) strongly inhibited the ability to stimulate SRE luciferase activity, similar to the lack of signaling by the Gαq-QL-C9,10S palmitoylation-deficient mutant and the loss of signaling upon treatment of cells expressing Gαq-QL with YM. As expected, the CA Gαq-QP and Gαq-RC mutants also robustly stimulated SRE luciferase activity, while the Gαq-I25A-QP and Gαq-I25A-RC mutants completely failed to stimulate luciferase activity, similar to cells treated with YM (E). The decreased signaling by Gαq-I25A-QL in comparison to Gαq-QL is much more drastic in the SRE luciferase assay than the pERK immunoblot assay (C and D). Although this is somewhat surprising since the SRE luciferase readout is downstream of the MAPK pathway, we note that the SRE luciferase assay provides a very high signal-to-noise readout for Gαq-QL signaling  and is consistently very sensitive to disruption of Gαq-QL signaling . Taken together, these results indicate that the introduction of the I25A mutation to disrupt binding to Gβγ inhibited oncogenic signaling of the CA Gαq mutants through the YAP and MAPK pathways, and the Gαq-QP and Gαq-RC mutants are more sensitive to the I25A mutation than Gαq-QL.Along with stimulation of the YAP pathway, CA Gα mutants , C and D mutants , C and Dactivity E. As ind with YM E. The deot assay , C and Dq-QL has an increased association with Gβγ compared to Gαq-RC  and was further quantified (B). As expected, WT Gαq displayed a strong pull down with Gβ1γ2, but the introduction of the I25A mutation into WT Gαq strongly decreased this interaction, indicating that the I25A mutation disrupts binding between Gαq and Gβγ. Interestingly, Gαq-QL had a similar binding association to Gβ1γ2 with that of WT Gαq, and Gαq-QL bound significantly stronger to Gβ1γ2 than Gαq-QP and Gαq-RC (A and B). Furthermore, Gαq-I25A-QL was more strongly bound to Gβ1γ2 than Gαq-I25A-QP and Gαq-I25A-RC (A and B). We did not see a statistically significant difference in association with Gβγ between Gαq-QP and Gαq-RC and their corresponding I25A mutants. This is likely due to the low initial binding to Gβγ with both Gαq-QP and Gαq-RC. Although we previously demonstrated the surprising ability of Gαq-QL to interact strongly with Gβγ and the poor interaction of Gαq-RC with Gβγ, we now show a dramatically decreased association of Gαq-QP with Gβγ compared to Gαq-QL, even though both have a mutation of Q209. This differential binding to Gβγ between the CA Gαq mutants likely contributes to the varying sensitivity in oncogenic signal disruption between the I25A mutants . Interestingly, cotransfection with 200 ng of the Gαo DNA was required to decrease Gαq-QL stimulation of TEAD luciferase activity by 63%. In comparison, cotransfection with 100 ng of Gαo plasmid DNA achieved 83% and 84% reduction in signaling by Gαq-QP and Gαq-RC, respectively. Transfection with the control Gαo G2A constructs did not reduce the luciferase reporter activity in response to the CA Gαq mutants (A). The ability of the expression of Gαo to reduce CA Gαq-stimulated YAP activity was also monitored through nuclear localization of YAP by immunofluorescence microscopy. Gαo localized strongly at cellular membranes, while the Gαo G2A mutant was primarily cytoplasmic, indicating the expected localization of the proteins. Strong nuclear YAP was detected with the CA Gαq-QL, Gαq-QP, and Gαq-RC mutants, indicating stimulation of the YAP pathway (B). As expected, expression of Gαo G2A with the CA Gαq mutants did not disrupt nuclear localization of YAP. Overexpression of Gαo inhibited YAP nuclear localization stimulated by Gαq-QP and Gαq-RC (B). Conversely, cotransfection of Gαo with Gαq-QL failed to prevent strong YAP nuclear localization (B), further suggesting that the Gαq-QP and Gαq-RC mutants are more sensitive to Gβγ binding disruption than Gαq-QL.Considering that inhibiting the association between the CA Gαin cells , 42, 43.embranes . YAP actplasmids A. Intere mutants A. The ab pathway B. As expd Gαq-RC B. Converlization B, furtheo expression and subsequent Gβγ sequestration on CA Gαq-promoted MAPK activity, we immunoblotted for pERK levels (A and B). The expression of the CA Gαq mutants resulted in strong stimulation of pERK. Expression of Gαo resulted in no significant decrease in pERK activity for Gαq-QL. However, cotransfection of Gαq-QP or Gαq-RC with 50 ng and 100 ng of Gαo plasmid resulted in a strong decrease in pERK activity (A and B). There was no significant change in pERK levels with the Gαo G2A control (A and B). These results were further validated using the SRE luciferase reporter assay (C). Expression of Gαq-QL, Gαq-QP, and Gαq-RC robustly activated SRE-dependent luciferase activity, while treatment with YM abolished luciferase activity. Consistent with results from the other signaling assays (A and B), cotransfection with 50 ng Gαo plasmid was sufficient to completely abolish Gαq-QP– and Gαq-RC–stimulated SRE luciferase activity, while up to 250 ng of Gαo plasmid was required for near complete inhibition of Gαq-QL–stimulated SRE luciferase activity (C). These findings indicate that the expression of Gαo to sequester endogenous Gβγ inhibits the signaling activity of Gαq-QL/P and Gαq-RC. The results also further validate that the Gαq-QL CA mutant is less sensitive to signaling disruption than the Gαq-QP and Gαq-RC mutants.To monitor the effects of GαK levels , A and Bactivity , A and B control , A and Ber assay C. Expresg assays , A and Bactivity C. These o could sequester Gβγ and inhibit the binding of Gβγ to Gαq-QL/P and Gαq-RC. To examine this, we coexpressed Gαo or Gαo G2A with the CA Gαq mutants in 6x-His-Gβ1y2 HEK 293 stable cells and pulled down Gβ1γ2 with Ni-NTA beads. Immunoblots were used to detect Gαq and Gαo bound to Gβ1 (A and B). As indicated in q-QL had a stronger binding association with Gβ1 than Gαq-QP and Gαq-RC. Gαo was bound to Gβ1 in the pull-down extract, but Gαo G2A had minimal to no interaction with Gβ1 as expected. The expression of Gαo resulted in significantly decreased binding of Gαq-QL to Gβ1 (A and B). Expression of Gαo consistently resulted in a noticeable decrease in the pull down of Gαq-QP and Gαq-RC with Gβγ, but this did not reach statistical significance due to the already low level of Gαq-QP and Gαq-RC association with Gβγ in the absence of Gαo expression. These findings confirm that overexpression of Gαo sequesters Gβγ and prevents binding of Gαq-QL/P and Gαq-RC to Gβγ.Next, we wanted to confirm that the expression of Gαd to Gβ1 , A and BL to Gβ1 , A and Bq through the N-terminal I25A mutation or expression of Gαo is sufficient to inhibit overactive Gαq signaling in HEK 293 Gαq/11 KO cells. We next wanted to determine if inhibiting Gβγ binding to CA Gαq would be sufficient to disrupt oncogenic signaling in UM cells containing the Gαq-QL or Gαq-QP mutants. To study this, we depleted Gβ1 and Gβ2, two predominant Gβ subunits, through siRNA transfections. Sufficient knockdown of Gβ1 and Gβ2 was first validated in HEK 293 Gαq/11 KO cells (A and B). siRNA molecules specific for Gβ1, Gβ2, and both Gβ1 and Gβ2 were transfected into the HEK 293 Gαq/11 KO cells, and WT Gαq, Gαq-QL, or Gαq-QP constructs were transfected after 24 h. pERK levels were detected and quantified through immunoblotting (A and B). The results indicate that siRNA depletion of Gβ1/2 significantly disrupted pERK activity driven by both Gαq-QL and Gαq-QP (A and B). Gβ1 or Gβ2 siRNA knockdown alone was insufficient to significantly inhibit pERK activation (A and B). Considering the effectiveness of Gβ1/2 depletion on pERK activity in the HEK 293 Gαq/11 KO cells, we decided to knock down Gβ1/2 in four UM cell lines: 1) Mel202 (Gαq-Q209L mutation), 2) 92.1 (Gαq-Q209L mutation), 3) OMM1.3 (Gαq-Q209P mutation), and 4) UM001 (Gαq-Q209P mutation). We also used the OCM-3 cell line as a control. The oncogenic activity of OCM-3 cells is driven by mutant BRAF (V600E) as opposed to mutant Gαq/11; thus, we would expect Gβ1/2 depletion to have minimal effect on pERK activity  and quantified (D). Interestingly, the OMM1.3 and UM001 cell lines which contain the Gαq-Q209P mutation had significantly decreased pERK levels with Gβ1/2 knockdown, similar to that of YM treatment (C and D). There was no significant difference in pERK levels with Gβ1/2 depletion in the Mel202 and 92.1 cells which contain the Gαq-Q209L mutation (C and D). This data further suggests that there is differential sensitivity to the inhibition of oncogenic signaling between cell lines with the Gαq-Q209L and Gαq-Q209P mutations.Our studies demonstrate that disrupting the interaction between Gβγ and CA GαKO cells , A and Bblotting , A and Bd Gαq-QP , A and Btivation , A and Bactivity . All celoblotted C and quaantified D. Interereatment , C and Dmutation , C and Dq and show a surprising differential sensitivity of CA Gαq mutants to the inhibition of signaling upon disruption of their interaction with Gβγ. We used two methods to disrupt Gβγ interaction with CA Gαq—introduction of a Gβγ-binding disrupting mutation, I25A, into CA Gαq and sequestration of endogenous Gβγ by expression of Gαo—to show that inhibiting interaction with Gβγ can drastically reduce CA Gαq-stimulated signaling to both the MAPK and YAP/TAZ pathways. Moreover, signaling by CA Gαq-Q209P, as well as CA Gαq-R183C, was much more effectively prevented by Gβγ binding disruption than signaling by CA Gαq-Q209L. Lastly, we extended the analysis to UM cell lines containing either the oncogenic driver mutant Gαq-Q209L or Gαq-Q209P and showed that depletion of Gβ1/2 strongly inhibited activation of the MAPK pathway in cells with the Gαq-Q209P mutant but not in cells with the Gαq-Q209L. Our results suggest that disrupting the interaction between Gβγ and CA Gαq may be a novel approach for inhibiting aberrant Gαq/11 signaling.The results presented in this study demonstrate the importance of Gβγ for signaling by CA mutants of Gαq mutants studied herein, it would be expected that CA Gα mutants would not retain binding to Gβγ. However, several studies have challenged the classical heterotrimer dissociation model, indicating that at least some Gα subunits maintain association with Gβγ when they are in the active, GTP-bound state due to the presence of a GTPase-inhibiting mutation or in response to GPCR activation  a central switch region, which undergoes a conformational change upon GTP binding and 2) an N-terminal α helical domain . Thus, one reason why CA Gαq mutants in this study showed decreased or loss of signaling to the MAPK and YAP pathways when interaction with Gβγ is disrupted is a failure to be efficiently palmitoylated and membrane-bound. Indeed, Gαq-I25A-QL retains stronger plasma membrane localization than Gαq-I25A-QP and Gαq-I25A-RC , although only partial loss of signaling was observed in when measuring pERK (C and D). Expression of the increasing amounts of Gαo to sequester Gβγ provided a powerful tool to demonstrate titratable differences in sensitivity to disruption of Gβγ binding to the CA Gαq mutants . Using the SRE-luciferase assay, transfection with 50 ng of Gαo expression plasmid was sufficient to completely abolish signaling of Gαq-QP and Gαq-RC, while transfection with 250 ng of Gαo plasmid was required to observe a similar almost-complete inhibition of Gαq-QL signaling (C). Likewise, transfection with 50 or 100 ng of Gαo plasmid failed to inhibit Gαq-QL–stimulated pERK levels, but transfection with 50 or 100 ng of Gαo plasmid provided significant inhibition of Gαq-QP– and Gαq-RC–stimulated pERK (A and B). Gβγ pull-down experiments showed the interaction of Gαq-QL with Gβγ, similar to the levels of WT Gαq interaction with Gβγ, and, importantly, the I25A mutation and expression of Gαo only partially disrupted the interaction . The Gαq-I25A, Gαq-I25A-QL, and Gαq-I25A-RC constructs were previously described . YFP-tagged Gαq was provided by Catherine Berlot. Gαo and Gαo-G2A plasmids were described previously (q and Gαo transient transfections was obtained from Invitrogen (Cat # 11668-019). siRNA targeting Gβ1 (GGAUAACAUUUGCUCCAUU), Gβ2 (ACUGGGUACCUGUCGUGUU), and both Gβ1 and Gβ2 (Gβ1/2) (ACGACGACUUCAACUGCAA) were previously described  (for immunoblots with HEK 293 Gαq/11 KO lysates), GAPDH (Cat # 60004-1-Ig), and Gαo (Cat # 12635-1-AP) were from Proteintech. The Gαq (Cat # ab199533) (for immunofluorescence experiments), Gβ1 (Cat # ab137635), and Gβ2 (Cat # ab108504) antibodies were obtained from Abcam. The YAP antibody (Cat # sc-101199) was purchased from Santa Cruz. The Gαq (Cat # 14373S) , ERK (Cat # 4696S), pERK (Cat # 9101S), and Myc-tag (Cat # 2272S) antibodies were obtained from Cell Signaling Technologies. The HA-tag antibody 12CA5 was from Covance. The GRK4-6 antibody (Cat # 05-466) was obtained from Sigma-Aldrich. For immunofluorescence microscopy, the secondary antibodies Alexa Fluor 488 (goat anti-rabbit) (Cat # A-11034), Alexa Fluor 594 (goat anti-mouse) (Cat # A-11032), Alexa Fluor 594 (goat anti-rabbit) (Cat # A-11037), and Alexa Fluor 647 (goat anti-mouse) (Cat # A-32728) were purchased from Invitrogen. The secondary antibodies IRDye 680RD goat anti-rabbit IgG (H + L) (Cat # 92568071) and IRDye 800CW donkey anti-mouse IgG (H + L) (Cat # 92532212) were obtained from LI-COR and were used to visualize protein from all of the immunoblots.The antibodies for Gαq/11 KO cells were generously provided by Dr Asuka Inoue and were described previously  with 10% fetal bovine serum (FBS)  and 1% penicillin/streptomycin . HEK 293 Gβ1γ2 stable cells were described previously . The SRE luciferase reporter plasmid was described previously  with 1% Triton X-100. For the Gαq-I25A experiments, the cells were incubated with the anti-rabbit Gαq antibody (Abcam) and the anti-mouse YAP antibody in 2.5% milk/TBS-Triton X-100. For the Gαo experiments, the cells were incubated with the anti-rabbit Gαo antibody and the anti-mouse YAP antibody in 2.5% milk/TBS-Triton X-100. The primary antibodies were incubated for 60 min. The cells were washed 5 times with 2.5% milk/TBS-Triton X-100. For the Gαq-I25A experiments, the cells were incubated with goat anti-rabbit Alexa Fluor 488 and goat anti-mouse Alexa Fluor 594 antibodies in 2.5% milk/TBS-Triton X-100. For the Gαo experiments, the cells were incubated with goat anti-rabbit Alexa Fluor 594 and goat anti-mouse Alexa Fluor 647 antibodies in 2.5% milk/TBS-Triton X-100. The secondary antibodies were incubated for 30 min. The cells were washed 5 times in TBS/1% Triton X-100. The cells were incubated in DAPI  diluted in warmed PBS for 5 min. The coverslips were rinsed in distilled water and mounted onto glass slides with ProLong Diamond Anti-fade Mountant . The images were acquired using the Olympus IX83 microscope with a 60× oil immersion objective and an ORCA Fusion sCMOS camera (Hamamatsu) controlled by Olympus cellSens software.HEK 293 Gα1γ2 pull-down assay was done as previously described . The cell lysates were incubated for 1 h on ice and were then centrifuged at 13,000 rpm  to pellet the nuclei and insoluble material. Forty microliters of the lysate were reserved separately for the input fraction. The remaining lysate was added to 30 μl of Ni-NTA beads  and rotated in an end-over-end rotator for 2 h at 4 °C. The tubes were placed in a magnetic rack, and the remaining supernatant was aspirated. The beads were washed 3 times with lysis buffer C. Fifty microliters of elution buffer (lysis buffer C with 0.25 M Imidazole) were added to elute Gβ1 and any bound proteins from the Ni-NTA beads. Forty microliters of the pull-down eluate were transferred to a new tube. Ten microliters of 5× SDS-PAGE sample buffer with 3.5% β-mercaptoethanol were added to the input fraction and the pull-down fraction. The input and pull-down lysates were separated on a 10% SDS-PAGE gel and protein bound was detected via immunoblotting. The blots were probed with the HA-tag and Myc-tag antibody . The blots for the Gαo experiments were also probed with Gαo and GAPDH. The blots were imaged on LI-COR, and the bands were quantified on the ImageJ software (https://imagej.nih.gov/ij/). The pull-down samples in Gαq-I25A experiments were normalized to its respective input fraction. The pull-down samples in the Gαo experiments were normalized to WT Gαq.The Gβescribed . Brieflyq  for the HEK 293 Gαq/11 KO cells or Cell Signaling Gαq  for the UM cells, along with GAPDH  as a loading control. The ERK and pERK bands were quantified by densitometry using the ImageJ software, and the relative pERK signal was divided by ERK. The quantified signals were normalized to WT Gαq (D), the individual CA Gαq mutants (B), or control siRNA between the CA Gαq mutants (B) or the UM cell lines (D).Cells were lysed in SDS-PAGE sample buffer. Lysates were run on a 10% SDS-PAGE gel and transferred to a nitrocellulose membrane. The membrane was blocked in 2.5% bovine serum albumin (BSA)  in 1× TBS supplemented with 0.05% Tween 20. The blots were incubated with both pERK  and ERK  antibodies overnight. A duplicate immunoblot was performed and was probed with either Proteintech Gαo WT Gαq D, the in mutants B, or con mutants B or the ll lines D.q/11 KO cells and 12-well plates for the UM cells. For the HEK 293 Gαq/11 KO experiments, 30 pmol of control, Gβ1, Gβ2, or Gβ1/2 siRNA were transfected into cells using Lipofectamine RNAiMax , according to manufacturer's instructions. The media was changed after 5 h. After 24 h, the corresponding WT Gαq, Gαq-QL, or Gαq-QP constructs were transfected into the cells using Lipofectamine 2000 . The media was changed to serum-free media after 24 h and corresponding cells were treated with 1 μM of YM. The cells were lysed in 1× SDS-PAGE sample buffer with 0.7% β-mercaptoethanol after 16 h. For experiments with the UM cells, 15pmol of control or Gβ1/2 siRNA were transfected into cells using Lipofectamine RNAiMax . The media was changed after 5 h. After 72 h, the media was changed to serum-free media, and the corresponding cells were treated with 1 μM of YM. The cells were further lysed after approximately 16 h in 1× SDS-PAGE sample buffer with 0.7% β-mercaptoethanol. The lysates were run on 10% SDS-PAGE gels and were immunoblotted for pERK , ERK , Gβ1 , Gβ2 , GAPDH , and Proteintech Gαq (Cat #13927-1-AP) for the HEK 293 Gαq/11 KO cells or Cell Signaling Gαq (Cat # 14373S) for the UM cells.Cells were plated in 6-well plates for the HEK 293 GαProtein lysates were run on 10% SDS-PAGE gels and transferred to LI-COR nitrocellulose membranes (Cat # nc9680617). The membranes were blocked in either 2.5% BSA or 2.5% milk in 1× TBS/0.05% Tween 20 at room temperature for 60 min. The blots were incubated in corresponding primary antibodies in 2.5% BSA or milk in 1× TBS/0.05% Tween 20 at 4 °C overnight. The blots were washed three times in 1× TBS/0.05% Tween 20 and incubated in LI-COR anti-rabbit (Cat # 92568071) and anti-mouse (Cat # 92532212) secondary antibodies for 60 min at room temperature. The immunoblots were further washed three times in 1× TBS/0.05% Tween 20 and once with PBS. The blots were then imaged on the LI-COR Odyssey imager.B and D) multiple comparison test were used to calculate significance. Error bars in all experiments indicate mean ± SD with significant differences indicated as ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.005; ∗∗∗∗p < 0.0001.GraphPad Prism was used to analyze the data for all of the figures. A two-way ANOVA followed by Šidák's or Tukey’s , B and DAll data is contained within the article.This article contains The authors declare that they have no conflicts of interest with the contents of this article.


The synthesis and crystal structure of a new Schiff base, namely, 4-bromo-2-[({2-[(2-hydroxyethyl)amino]ethyl}imino)methyl]phenol, is reported. 11H15BrN2O2, crystallizes in the monoclinic space group P21 with two independent molecules in the asymmetric unit. It was prepared by the condensation reaction of 5-bromo-2-hydroxybenzaldehyde and aminoethylethanolamine. There is an intramolecular O—H⋯N hydrogen bond with an S(6) ring motif. Moreover, there are intermolecular C—H⋯N, C—H⋯O and Br⋯O interactions in the crystal structure of the title compound.The new title Schiff base compound, C Moreover, the O2a and O2b atoms are involved in a second intramolecular hydrogen bond. The molecules are connected through intermolecular O—H⋯N hydrogen bonds and Br⋯O interactions with distances Br1a⋯O2b = 3.206 (2) Å and Br1b⋯O2a = 3.282 (2) Å  was dissolved in ethanol (10 ml) and stirred for 10 min. Then, a solution of aminoethylethanolamine (0.2 mmol) dissolved in ethanol (5 ml) was added dropwise. The mixture was stirred and refluxed for 6 h. After that, the solution was concentrated under reduced pressure. Yellow crystals suitable for X-ray analysis were obtained by slow evaporation of solvent at room temperature for several days. These were filtered off and washed several times with cold ethanol.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2414314621003357/bt4109sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2414314621003357/bt4109Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314621003357/bt4109Isup3.cmlSupporting information file. DOI: 2074082CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


II ion of the complex cation has an N3O square-planar coordination sphere defined by the three N atoms of the tridentate 2,2′:6′,2′′-terpyridine ligand and one O atom from the NO3− anion.The central Pd 3)(C15H11N3)]NO3, comprises a cationic PdII complex and a nitrate anion. In the complex, the PdII cation is four-coordinated in a distorted square-planar coordination geometry defined by the three N atoms of the tridentate 2,2′:6′,2′′-terpyridine ligand and one O atom from the NO3− anion. In the crystal, the complex molecules are stacked in columns along the a axis being connected by π–π stacking [closest inter-centroid separation between pyridyl rings = 3.878 (3) Å]. The connections between columns and anions to sustain a three-dimensional architecture are C—H⋯O hydrogen bonds.The title complex, [Pd(NO The Pd—N [1.917 (4) to 2.030 (4) Å] and Pd—O [2.028 (3) Å] bond lengths are close. The pyridine rings of the terpy ligand are located approximately parallel to the least-squares plane of the PdN3O unit [maximum deviation = 0.023 (2) Å], with dihedral angles of 1.4 (2)° (ring N1/C1–C5), 3.1 (2)° (ring N2/C6–C10) and 3.0 (2)° (ring N3/C11–C15). In the crystal  and Cg2i , the centroid-centroid distance is 3.878 (3) Å and the dihedral angle between the ring planes is 3.2 (3)° 2·2H2O  in acetone (30 ml) was added 2,2′:6′,2′′-terpyridine  followed by stirring for 3 h at room temperature. The formed precipitate was separated by filtration, washed with acetone and dried at 323 K to give a light-yellow powder (0.2123 g). Yellow crystals of the product suitable for X-ray analysis were obtained by slow evaporation of its CH3NO2 solution at room temperature.To a solution of Pd I. DOI: 10.1107/S2414314621000857/tk4067Isup2.hklStructure factors: contains datablock(s) I. DOI: 2058389CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


P-substituent benzoate anions or a trifluoroacetate anion. They are hydrated and there are three crystallized as 1:1 salts while the fourth is a 2:2 salt. Their crystal packing depends on strong ribbons or sheets stabilized by hydrogen bonds of type N—H⋯O and O—H⋯O and other interactions as C—H⋯O, C—H⋯π and C—H⋯F in the trifluoroacetate-based one.Four novel piperazinium salts are reported, and based on 1-phenylpiperazinium as a common cation in the asymmetric units that include additionally water molecules, and different  9H9O3·C10H15N2·H2O (I); 4-phenylpiperazin-1-ium 4-methoxybenzoate monohydrate, C10H15N2·C8H7O3·H2O (II); 4-phenylpiperazin-1-ium 4-methylbenzoate monohydrate, C10H15N2·C8H7O2·H2O (III); and 4-phenylpiperazin-1-ium trifluoroacetate 0.12 hydrate, C10H15N2·C2F3O2·0.12H2O (IV), have been synthesized. The single-crystal structures of these compounds reveal that all of them crystallize in the triclinic PI)–(III) is built up of ribbons formed by a combination of hydrogen bonds of type N—H⋯O, O—H⋯O and other weak interactions of type C—H⋯O and C—H⋯π, leading to a three-dimensional network. In the crystal of (IV), the cations and the anions are connected by C—H⋯O, N—H⋯O and C—H⋯F hydrogen bonds and by C—H⋯π interactions, forming sheets which in turn interact to maintain the crystal structure by linking through the oxygen atoms of water molecules and van der Waals interactions, giving the whole structure.In this study, four new piperazinium salts, namely, 4-phenylpiperazin-1-ium 4-ethoxybenzoate monohydrate, C In compound (II) the asymmetric unit  Å, θ = 175.89 (17)° and φ = 346 (3)°. Compound (III) presents an asymmetric unit  Å, θ = 9.38 (19)° and φ = 167.9 (13)°. On the other hand, the asymmetric unit of (IV)  and two trifluoroacetate anions  and a 0.12 occupancy water molecule. The aromatic rings of the cations  are essentially planar while the protonated piperazine rings adopt a chair conformation for cation A1, with puckering parameters  Å, θ = 0.0 (4)° and φ = 207 (14)°, and a distorted chair conformation for the cation A2, with puckering parameters QT = 0.559 (5) Å, θ = 6.6 (4)° and φ = 168 (4)°.The asymmetric unit of the compound , Fig. 1, consist), Fig. 1 QT = 0.it Fig. 2 compriseit Fig. 3 composedV) Fig. 4 contains3.I), the cation pairs are connected across two water molecules by C—H⋯O and N—H⋯O hydrogen bonds, forming an a-axis direction (Table 1a). In addition, a set of C—H⋯π interactions, through the benzene rings of the anions and the cations, connect the molecules together in ribbons along the a-axis direction (Table 1b). The C—H⋯O, N—H⋯O, O—H⋯O hydrogen bonds and C—H⋯π interactions together form a three-dimensional network, contributing to the stabilization of the crystal structure.In the crystal structure of (n Table 1a. In adn Table 1b. The CII), the cations, the anions and the water molecules are connected by C—H⋯O, N—H⋯O and O—H⋯O hydrogen bonds, forming ribbons along the a-axis direction (Table 2a). Furthermore, the cations interact via C—-H⋯π interactions through the benzene ring of the anion, forming ribbons along the b-axis direction (Table 2b). The C—H⋯O, N—H⋯O, O—H⋯O hydrogen bonds and C—H⋯π interactions together form a three-dimensional network, contributing to the stabilization of the crystal structure.In the crystal structure of (n Table 2a. Furthn Table 2b. The CIII), the cations, the anions and the water molecules are connected by C—H⋯O, N—H⋯O and O—H⋯O hydrogen bonds, forming ribbons along the a-axis direction , the cations and the anions are connected by C—H⋯O, N—H⋯O and C—H⋯F hydrogen bonds (Table 4a) and C—H⋯π interactions, generating sheets parallel to the (100) plane (Table 4IV) involve the oxygen atoms of carboxylate groups, while the 0.12 fraction of the water molecule contributes with one interaction of the type C—H⋯O and it is weak in comparison to the other oxygen-based ones.In the crystal structure of –(III) gave no hits. However, searching for a branched phenyl piperazinium cation and para-substituted benzoate anion gave comparable hits, namely; 4-(4-methoxyphenyl)piperazin-1-ium 4-fluorobenzoate monohydrate, 4-(4-methoxyphenyl)piperazin-1-ium 4-chlorobenzoate monohydrate, 4-(4-methoxyphenyl)piperazin-1-ium 4-bromobenzoate monohydrate –(IV) have no substituent. They also crystallize as monohydrates, and their crystal structures are based on differently sized chains of rings formed via a combination of hydrogen bonds of the type N–H⋯O and O–H⋯O and other weak interactions of types C—H⋯O and C—H⋯π to form sheets. In 4-(4-methoxyphenyl)piperazin-1-ium 4-aminobenzoate monohydrate  gave no hits.A search of the Cambridge Structural Database –(IV), a solution of commercially available (from Sigma–Aldrich) 1-phenylpiperazine  in methanol (10 ml) was mixed with equimolar solutions of the appropriate organic acids in methanol (10 ml) viz., 4-ethoxybenzoic acid  for (I), 4-methoxybenzoic acid  for (II), 4-methylbenzoic acid  for (III) and trifluoroacetic acid  for (IV). The corresponding solutions were stirred for 15 min at room temperature and allowed to stand at the same temperature. X-ray quality crystals were formed on slow evaporation in a week for all compounds, where ethanol:ethylacetate (1:1) was used for crystallization. The corresponding melting points were 353–355 K for (I), 368–370 K for (II), 338–340 K for (III) and 385–387 K for (IV).For the synthesis of salts (6.Uiso(H) = 1.2Ueq(C) or 1.5Ueq(C). For the H atoms bonded to the N and O atoms, the atomic coordinates were refined with Uiso(H) = 1.2Ueq(N) and 1.5Ueq(O), . In (IV), the atoms of the CF3 groups of two trifluoroacetate anions  are disordered over two sets of sites with site occupancies of 0.737 (3) and 0.263 (3). The corresponding bond distances in the disordered groups were restrained to be equal. The Uij components of these atoms were restrained to be equal and were restrained to approximate isotropic behaviour. The OW1 water molecule was refined with a resulting occupation factor of 0.245 (10) and the H atoms of the water molecule were placed geometrically.Crystal data, data collection and structure refinement details are summarized in Table 510.1107/S2056989022006004/dj2048sup1.cifCrystal structure: contains datablock(s) global, I, II, III, IV. DOI: 10.1107/S2056989022006004/dj2048Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022006004/dj2048Isup6.cmlSupporting information file. DOI: 10.1107/S2056989022006004/dj2048IIsup3.hklStructure factors: contains datablock(s) II. DOI: Click here for additional data file.10.1107/S2056989022006004/dj2048IIsup7.cmlSupporting information file. DOI: 10.1107/S2056989022006004/dj2048IIIsup4.hklStructure factors: contains datablock(s) III. DOI: Click here for additional data file.10.1107/S2056989022006004/dj2048IIIsup8.cmlSupporting information file. DOI: 10.1107/S2056989022006004/dj2048IVsup5.hklStructure factors: contains datablock(s) IV. DOI: Click here for additional data file.10.1107/S2056989022006004/dj2048IVsup9.cmlSupporting information file. DOI: 2177037, 2177036, 2177035, 2177034CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


A new organic salt of diclofenac, [2-phenyl]acetate, with monoethanolammonium and water has been obtained, and its structure has been established by X-ray analysis. 2H8NO+·C14H10Cl2NO2−·H2O, the asymmetric unit contains one cation, one anion and a water molecule, all in general positions. A complex network of hydrogen bonds is present in the crystal structure.In the solid-state structure of the title compound derived from diclofenac, C The dicb-axis direction, due to the crystallographic twofold screw axis, via N2—H2B⋯O1Wii hydrogen bond [2.947 (4) Å, symmetry code: (ii) −x, y − z + c [O1W⋯O1iv, symmetry code: (iv) x, −y + z + i, symmetry code: (i) −x, −y + 1, −z + 1, 2.811 (4) Å, Fig. 2X⋯Cg π–ring interactions, for C3—H3 and C7—Cl1 bonds, for which the X⋯Cg separations and γ angles range from 3.533 to 3.958 Å and from 25.03 to 28.79°.The ionic form of the title compound serves as a building block for the supramolecular architecture. In the crystal, the building blocks form screw-like chains along the Crystal Explorer 17.5  at 298 K for 5 min. The solution was then placed in a loosely closed bottle and kept at 298 K for 10 days. The precipitated prismatic crystals were selected for the single-crystal X-ray diffraction analysis.To a solution of 0.1 g (0.52 mmol) of D in 4 ml of ethanol, 32 µCrystal data, data collection and structure refinement details are summarized in Table 210.1107/S2414314622004412/bh4068sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314622004412/bh4068Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2414314622004412/bh4068Isup3.cmlSupporting information file. DOI: 2168795CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


I atom of the cationic complex of the title compound, [Ir(C8H12)(C18H15P)(C6H11N3)][BF4] ·0.8CH2Cl2, exhibits a distorted square-planar coordination environment.The central Ir 8H12)(C18H15P)(C6H11N3)]BF4·0.8CH2Cl2, has been synthesized and structurally characterized. The central IrI atom of the cationic complex has a distorted square-planar coordination environment, formed by a bidentate cycloocta-1,5-diene (COD) ligand, an N-heterocyclic carbene, and a triphenylphosphane ligand. The crystal structure comprises C—H⋯π(ring) interactions that orient the phenyl rings; non-classical hydrogen-bonding interactions between the cationic complex and the tetrafluoridoborate anion are also present. The complex crystallizes in a triclinic unit cell with two structural units and an incorporation of dichloromethane solvate molecules with an occupancy of 0.8.A new triazole-based N-heterocyclic carbene iridium(I) cationic complex with a tetrafluoridoborate counter-anion, [Ir(C The distorted square-planar environment around the IrI atom is defined by the bidentate cycloocta-1,5-diene (COD) ligand, the carbene C1 atom of the triazole NHC ligand, and the P atom of the triphenylphosphane ligand. The P1—Ir1—C1 bond angle is 93.88 (10)°. The N1—C1—N3 bond angle of the coordinating carbene atom significantly differs with a value of 103.3 (3)° from the expected 2sp hybridization.The molecular structure of the title complex 4]− group. Additionally, there are non-classical intermolecular hydrogen-bonding interactions between the hydrogen atom of a phenyl group (H10) and a nitrogen atom of the NHC ligand (N2). Non-classical hydrogen bonding interactions are shown as dotted green lines in Fig. 2The crystal packing of the title compound is displayed in Fig. 2A) and a phenyl phosphane ring (C19–C24). This intramolecular interaction displays an H⋯centroid distance of 2.61 Å and a C—H⋯centroid angle of 168°. The intermolecular C—H⋯π(ring) interaction orients phenyl phosphane rings of adjacent moieties as it occurs between a hydrogen atom of a phenyl ring (H21) and an adjacent phenyl ring (C13–C18). The intermolecular C—H⋯π(ring) interaction has an H⋯centroid distance of 2.73 Å and a C—H⋯centroid angle of 157°. The C—H⋯π(ring) interactions orient phenyl rings on adjacent moieties (C13–C18 and C19–C24) into an approximately perpendicular arrangement, shown in Fig. 3Both intermolecular and intramolecular C—H⋯π(ring) interactions are observed and shown as dashed orange lines in Figs. 2 chloroiridium (1) was synthesized by a previously published procedure  Varian spectrometer and referenced to the residual solvent peak of CDCl3 (δ in p.p.m.).(triphenylphosphane)iridium(I) tetrafluoridoborate (2): Triphenylphosphane  and AgBF4 (0.048 g 0.245 mmol) were added to an oven-dried flask containing complex (1)  in 10 ml of CH2Cl2, and stirred under N2 in the dark for 90 min. The mixture was filtered through Celite and the solvent was removed under reduced pressure. The bright orange–red solid was washed with pentane and dried under vacuum yielding 0.165 g (86.9%) of the title compound 2. 1H NMR: δ (p.p.m.) 8.18 , 7.49–7.32 , 5.36 2), 4.38, 3.99 , 4.05 , 2.27–1.6 , 1.56 . 13C NMR: δ 177.74 (Ir—C), 140.32 (N—CH—N), 132.46–128.38 (Carom), 87.82, 87.43, 85.34, 85.01 (CH of COD), 53.23 [CH(CH3)2], 41.31 (N—CH3), 33.41, 33.18, 31.45, 30.39 CH2 of COD, 24.37, 22.15 [CH(CH3)2]. 31P: δ 17.23.2 was crystallized by slow diffusion of pentane into a CH2Cl2 solution.The title compound Crystal data, data collection, and structure refinement details are summarized in Table 210.1107/S2414314623000640/wm4181sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314623000640/wm4181Isup2.hklStructure factors: contains datablock(s) I. DOI: 2237810CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The syntheses and low-temperature crystal structures of four organic salts of 4-(4-nitrophenyl)piperazine are presented. 10H14N3O2+·C4H5O4− (I), 4-(4-nitrophenyl)piperazinium 4-aminobenzoate monohydrate, C10H14N3O2+·C7H6NO2−·H2O (II), 4-(4-nitrophenyl)piperazinium 2-(4-chlorophenyl)acetate, C10H14N3O2+·C8H6ClO2− (III) and 4-(4-nitrophenyl)piperazinium 2,3,4,5,6-pentafluorobenzoate, C10H14N3O2+·C7F5O2− (IV). The salts form from mixtures of N-(4-nitrophenyl)piperazine and the corresponding acid  in mixed solvents of methanol and ethyl acetate. Salts I, III, and IV are anhydrous, whereas II is a monohydrate. In each structure, the overall conformation of the cation is determined by the disposition of the exocyclic N—C bond of the piperazine ring  and twists about the N—C bond between the piperazine ring and its attached 4-nitrophenyl ring. The packing motifs in each structure are quite different, though all are dominated by strong N—H⋯O hydrogen bonds, which are augmented in I and II by O—H⋯O hydrogen bonds, and in III by a π–π stacking interaction between inversion-related 4-nitrophenyl groups.The syntheses and crystal structures are presented for four organic salts of the 4-(4-nitrophenyl)piperazinium cation, namely, 4-(4-nitrophenyl)piperazinium hydrogen succinate, C The relative twist about these N2—C5 bonds, e.g. quantified by the C2—N2—C5—C6 torsional angles  determine the overall cation shape. In each case, the 4-nitro group is essentially coplanar with its attached phenyl ring.The overall conformations of the 4-nitrophenylpiperazinium cations in s Figs. 1 ▸ ▸. WI has minor twists about its three C—C bonds , which leads to a dihedral angle of 34.63 (9)° between its carboxylate/carboxylic acid groups. The 4-aminobenzoate anion of II is close to planar, having a dihedral angle between the carboxylate group and its benzene ring of 10.70 (7)°. The amine group at N4 is also slightly non-planar [the sum of angles about N4 is 349 (2)°]. In the 2-(4-chlorophenyl)acetate anion of III, twists about the C11—C12 and C12—C13 bonds place the carboxylate group almost perpendicular [85.02 (9)°] to the benzene ring. Lastly, in the pentafluorobenzoate anion of IV, the carboxylate group is 55.95 (10)° out of coplanarity with the phenyl ring.The succinate anion in I, which will be described in more detail in the next section (Supramolecular features).Throughout all four structures, individual bond lengths and angles take on normal values except for an elongated O—H bond [1.17 (2) Å] in 3.2 group and the anionic carboxylate groups. In I, II, and IV, these hydrogen bonds to the anion are equatorial relative to the piperazine ring, while that in III is axial. Nevertheless, in each structure, the NH2+ group acts as a hydrogen-bond donor through both its hydrogen atoms. In I, III, and IV this is to a second anion, whereas in II it is to the included water molecule. Throughout the four structures, all conventional N—H⋯O and all but one O—H⋯O  hydrogen bonds take on normal distances and angles  hydrogen bond [O⋯O = 2.4367 (10) Å], which links adjacent hydrogen-succinate anions into chains that propagate parallel to the b-axis direction  anions and as an acceptor for an N—H⋯O hydrogen bond, generates a double-layer network lying parallel to (011)  Fig. 7. Of the III joins two pairs of inversion-related ammonium cations and carboxylate anions, forming an III also includes the only π–π interactions of the four structures, which occurs between inversion-related  nitrophenyl rings, giving an interplanar spacing of 3.3352 (15) Å, though the offset (≃1.92 Å) is large, leading to a centroid–centroid distance of 3.8495 (15) Å piperazine as a guest molecule  for I, 4-aminobenzoic acid  for II, 2-(4-chlorophenyl)acetic acid  for III, and 2,3,4,5,6-pentafluorobenzoic acid  for IV. The resulting solutions were stirred for 30 minutes at 333 K and allowed to stand at room temperature. X-ray quality crystals formed on slow evaporation of solutions in ethanol:acetonitrile (1:1) over the course of a week for all four compounds. The melting points are 398–400 K (I), 473–475 K (II), 431–435 K (III) and 411–415 K (IV).A solution of commercially available (Sigma-Aldrich) 4-nitrophenylpiperazine  in methanol (10 ml) was mixed with equimolar solutions of the appropriate acid in methanol (10 ml) and ethyl acetate (10 ml) 6.sp2—H) and 0.99 Å (R2CH2), using Uiso(H) values constrained to 1.2Ueq of the attached carbon atom. All N—H and O—H hydrogen atoms were refined freely (both coordinates and Uiso).Crystal data, data collection, and structure refinement details are given in Table 510.1107/S2056989023001093/hb8053sup1.cifCrystal structure: contains datablock(s) I, II, III, IV, global. DOI: 10.1107/S2056989023001093/hb8053Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989023001093/hb8053IIsup3.hklStructure factors: contains datablock(s) II. DOI: 10.1107/S2056989023001093/hb8053IIIsup4.hklStructure factors: contains datablock(s) III. DOI: 10.1107/S2056989023001093/hb8053IVsup5.hklStructure factors: contains datablock(s) IV. DOI: Click here for additional data file.10.1107/S2056989023001093/hb8053Isup6.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001093/hb8053IIsup7.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001093/hb8053IIIsup8.cmlSupporting information file. DOI: Click here for additional data file.10.1107/S2056989023001093/hb8053IVsup9.cmlSupporting information file. DOI: 2239906, 2239905, 2239904, 2239903CCDC references:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Correction: BMC Musculoskelet Disord 23, 630 (2022)https://doi.org/10.1186/s12891-022-05517-0Following the publication of the original article  the auth“Their mean age at surgery was 60.8 ± 10.3 years and the average symptom duration was 40.7 ± 34.1 months.”Should be corrected to:“Their mean age at surgery was 61.2 ± 10.8 years and the average symptom duration was 23.36 ± 22.11 months.”The original article  has been


II(C68H44N12O4)(C4H6N2)2]·C4H6N2·1.5C4H8O is investigated.The crystal structure of the bis(1-methylimidazole)-ligated iron(II) picket-fence porphyrin derivative [Fe II(C68H44N12O4)(C4H6N2)2]·C4H6N2·1.5C4H8O, the central FeII ion is coordinated by four pyrrole N atoms of the porphyrin core and two N atoms of the 1-methylimidazole ligands in the axial sites. One 1-methylimidazole and one and a half tetrahydrofuran solvent molecules are also present in the asymmetric unit. The complex exhibits a near planar porphyrin core conformation, in which the iron centre is slightly displaced towards the hindered porphyrin side (0.01 Å). The average Fe—Np (Np refers to the pyrrole nitrogen atoms in the porphyrin) bond length is 1.990 (9) Å, and the axial Fe—NIm (NIm refers to the imidazole nitrogen atoms) bond lengths are 1.993 (3) and 2.004 (3) Å. The dihedral angle between the two coordinated 1-methylimidazole planes is 56.6 (2)°. The dihedral angles between the 1-methylimidazole planes and the planes of the closest Fe—Np vector are 16.8 (2) and 39.8 (2)°. N—H⋯N and N—H⋯O interactions are observed in the crystal structure.In the title compound, [Fe Additional quantitative information on the structure is given in Fig. 2p vector are 16.8 (2) and 39.8 (2)°. The dihedral angle between the two coordinated imidazole planes is 56.6 (2)°, showing a relative perpendicular orientation. Fig. 2p—Fe—Np angle is ideal at 90.01 (9)°, and the axial Fe—NIm bond lengths are 1.993 (3) and 2.004 (3) Å. The average Fe—Np distance of 1.990 (9) Å is a typical value for low-spin ferrous porphyrin derivatives  was distilled from Na/benzophenone under N2. Hexanes were distilled over sodium/potassium alloy under N2. Solvents were degassed by repeated freeze–pump–thaw cycles. 1-MeIm was distilled under argon before use. Precursors H2TPyPP, [Fe(TPyPP)]Cl, and [Fe(TPyPP)]OH were prepared following literature methods , with slight modifications.Synthesis of the title compound. [Fe(TPyPP)]OH  and 1-MeIm  were dissolved in 3 ml of THF. The mixture was stirred for 15 min and transferred into glass tubes (8 mm × 10 cm), which were layered with hexanes. Several days later, X-ray quality black block-shaped crystals were collected.bCrystal data, data collection and structure refinement details are summarized in Table 210.1107/S2414314621005319/bh4061sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2414314621005319/bh4061Isup2.hklStructure factors: contains datablock(s) I. DOI: 2068473CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The study investigated clinical features of sellar and suprasellar tumors with optic nerve bending. Twenty-five patients  with optic nerve bending in one eye who underwent tumor resection for sellar and suprasellar tumors were included. The other eye, without optic nerve bending, was the control. The pre- and postoperative best-corrected visual acuity (BCVA) and ganglion cell layer (GCL) + inner plexiform layer (IPL) thickness were studied retrospectively using optical coherence tomography. Preoperative BCVA in the eye with optic nerve bending was significantly poor and improved significantly after tumor resection. Eyes with optic nerve bending had significantly less GCL + IPL thickness on the temporal side than eyes without optic nerve bending. Preoperative GCL + IPL thickness of the entire macula was reduced in eyes with optic nerve bending and poor postoperative BCVA compared to those with good postoperative BCVA. There was no significant difference in GCL + IPL thickness of eyes with optic nerve bending before and after tumor resection. Optic nerve bending caused by sellar and suprasellar tumors resulted in visual impairment and decreased retinal ganglion cells. Eyes with optic nerve bending and severely reduced GCL + IPL thickness may have less BCVA improvement after tumor resection. Visual field impairment progresses gradually as the tumor grows, resulting in visual impairment3. However, even a relatively slight compression of the optic chiasm can cause visual impairment. We investigated the visual acuity impairment in the patients with sellar and suprasellar lesions using magnetic resonance (MR) T2 weighted imaging4. We also reported a new concept that the optic nerve sometimes bends at the entrance of the optic canal from the intracranial subarachnoid space. Further, a combination of preoperative optic nerve bending and optic chiasm compression is associated with visual impairment  can measure ganglion cell layer (GCL) + inner plexiform layer (IPL) thickness in the macula with high reliability. Previous reports showed the usefulness of macular GCL + IPL parameters in discriminating early glaucoma from normal eyes5. Morphological changes in retinal ganglion cells might also be observed using OCT in patients with sellar and suprasellar tumors with optic nerve bending. Here, we investigated the OCT findings of patients with sellar and suprasellar tumors with optic nerve bending before and after tumor resection.Sellar and suprasellar tumors, including pituitary adenomas, show visual field defects, such as bilateral hemianopsia, due to the optic chiasm compression by the tumorTable All 25 patients with optic nerve bending were classified by age into groups of 30–40 s (n = 6), 50 s (n = 7), 60 s (n = 5), and 70 s (n = 7). Postoperative BCVAs (logMAR) were − 0.08 ± 0.00 (30–40 s),0.01 ± 0.17 (50 s), 0.08 ± 0.31 (60 s) and 0.13 ± 0.25 (70 s), respectively. Postoperative BCVA showed no significant differences among these 4 groups .The preoperative GCL + IPL thicknesses in bending and non-bending eyes were 65.6 ± 12.8 µm and 71.4 ± 9.7 µm in the superior sector, 64.8 ± 13.1 µm and 68.8 ± 11.1 µm in the superior nasal sector, 61.9 ± 12.4 µm and 64.8 ± 10.6 µm in the inferior nasal sector, 63.8 ± 12.9 µm and 70.0 ± 8.4 µm in the inferior sector, 72.9 ± 12.8 µm and 80.6 ± 11.5 µm in the inferior temporal sector, and 69.9 ± 11.8 µm and 75.8 ± 8.0 µm in the superior temporal sector, respectively. The postoperative GCL + IPL thicknesses in the bending and non-bending eyes were 65.2 ± 12.7 µm and 71.0 ± 9.1 µm in the superior sector, 63.6 ± 13.7 µm and 68.2 ± 10.7 µm in the superior nasal sector, 60.8 ± 12.6 µm and 64.6 ± 10.5 µm in the inferior nasal sector, 64.6 ± 11.4 µm and 68.4 ± 8.2 µm in the inferior sector, 72.3 ± 12.7 µm and 79.9 ± 12.3 µm in the inferior temporal sector, and 69.1 ± 12.4 µm and 75.6 ± 8.0 µm in the superior temporal sector, respectively  of 0 or better as good visual outcome and 6 eyes with postoperative BCVA (logMAR) of less than 0 as poor visual outcome. The preoperative GCL + IPL thicknesses in the good visual outcome eyes and the poor visual outcome eyes were 69.9 ± 8.1 µm and 52.0 ± 15.3 µm in the superior sector, 68.7 ± 10.5 µm and 52.3 ± 12.8 µm in the superior nasal sector, 65.7 ± 9.6 µm and 49.8 ± 12.5 µm in the inferior nasal sector, 68.9 ± 6.6 µm and 47.8 ± 14.6 µm in the inferior sector, 76.9 ± 6.7 µm and 60.0 ± 17.9 µm in the inferior temporal sector, and 73.6 ± 6.7 µm and 58.2 ± 16.0 µm in the superior temporal sector, respectively  analysis, of sellar and suprasellar tumors with sagittal bending of the optic nerve and compared them with those of non-bending optic nerve controls. Eyes with optic nerve bending due to sellar and suprasellar tumors had worse visual acuity and reduced GCL + IPL thickness in the temporal sectors, as measured by OCT, than eyes without optic nerve bending. In addition, eyes with optic nerve bending showed rapid improvement in visual acuity after tumor resection. Furthermore, in six eyes with poor visual outcome, the preoperative GCL + IPL thickness was significantly lesser than that in 19 eyes with good visual outcome.4. ONCBA basically affects ipsilateral vision4. Moreover, ipsilateral ONCBA is anatomically unrelated to contralateral visual dysfunction. However, when the ONCBA is large, the tumor is often large; therefore, the visual field defect due to chiasma compression may occur bilaterally. In addition, if the tumor is larger, the ONCBA on the contralateral side may be large. In this study, eyes with optic nerve bending had preoperative visual impairment, whereas eyes without optic nerve bending had good preoperative visual acuity , the preoperative OCT of the optic nerve bending eye showed only a mild decrease in GCL + IPL in the predominantly superior nasal sector, and the postoperative BCVA improved  made evaluations, and any disagreements regarding conclusions were resolved by consensus. Optic nerve bending (large ONCBA) was defined as ONCBA ≥ 45°, and non-optic nerve bending (moderate ONCBA) was defined as ONCBA < 45°, as previously reported4. The exclusion criteria were as follows: (1) patients with a history of glaucoma or evident glaucomatous optic neuropathy; (2) high myopia (refractive error less than − 6 diopters); (3) retinal diseases, including epiretinal membrane and macular edema; (4) severe cataract, and (5) unclear optic nerve on MR imaging.All experiments followed the tenets of the Declaration of Helsinki and were approved by the Institutional Review Board of the Gunma University Graduate School of Medicine (HS2021-097). Informed consent was obtained from all individual participants in the present study. We retrospectively studied 25 patients with visual impairment due to sellar and suprasellar tumors who underwent endoscopic transsphenoidal tumor resection at Gunma University Hospital from June 2015 to July 2021 and had optic nerve bending in only one eye. The other eye, without optic nerve bending, was used as the control. MR imaging of the sellar and suprasellar lesions in all 25 patients with a 1.5 T or 3 T MR imaging system was performed. The presence of optic nerve bending was determined by measuring the sagittal ONCBA on MR images before tumor resection, as previously reported5.All patients underwent ophthalmologic examinations, including best-corrected visual acuity (BCVA), intraocular pressure assessment, refraction, slit-lamp biomicroscopy, fundus examination, and GCL + IPL thickness measurement, using Cirrus high definition-OCT  in both the optic nerve bending and non-bending eyes before and 1 month after tumor resection Fig. . BCVA waData are presented as the mean ± standard deviation. An unpaired t-test was conducted to compare BCVA and GCL + IPL thickness measurements between the bending and non-bending eyes. The paired t-test was conducted to compare changes in BCVA and GCL + IPL thickness before and 1 month after surgery. The correction between BCVA and ONCBA was examined using Spearman’s correlation coefficient. Statistical significance was set at p < 0.05. Statistical analyses were performed using GraphPad Prism version 6 .


In the crystal, ion pairs are linked by C—H⋯Br and N—H⋯Br hydrogen bonds and are connected into helical chains extending along the c-axis direction by weak, electrostatic S⋯Br− interactions. A Hirshfeld surface analysis was performed, which showed the dominant role of H⋯H contacts (51.3%).In the title molecular salt, C The dihedral angle between the mean planes of the thiazole and C2–C7 rings is 13.1 (1)°. A puckering analysis of the C1/C2/C7–C10 ring yielded the parameters Q = 0.499 (3) Å, θ = 58.6 (3)° and φ = 225.6 (3)°, indicating a half-chair conformation.As expected, the C11/C12/C13/N3/S1 thiazole ring in − contacts of this length or shorter are present in the Cambridge Structural Database, two examples being reported by Auffinger et al. .In the crystal, the S1⋯Br1 distance of 3.5017 (7) Å is some 0.15 Å less than the sum of the van der Waals radii and likely represents an electrostatic interaction between the two atoms since S1 is near to the cationic charge. Over 200 structures having S⋯Br al. 2004 and Thom al. 2004. This ins Table 1 form helon Fig. 2. It may ts Fig. 3, in agreet al., 2016A yielded 30 hits of which 11 were considered similar to I. Among these, (Z)-1--2-[1-(4-hydroxyphenyl)ethylidene]hydrazinium bromide unknown solvate -2-[(2-nitrophenyl)methylidene]-1-hydrazinium bromide -ylidene)hydrazinylidene]ethyl}pyridinium bromide monohydrate . Fig. 4a presents the Hirshfeld surface plotted over dnorm with a second cation closest to the bromide ion also present, clearly showing the N—H⋯Br and C—H⋯Br interactions as well as the S1⋯Br1 short contact (dashed lines). The surface plotted over shape  and curvature indices  do not show much flat surface or evidence for π-stacking interactions, in agreement with the results given in Section 3. Fig. 5a) and resolved into all H⋯H contacts , H⋯C/C⋯H contacts , Br⋯H/H⋯Br contacts  and S⋯H/H⋯S contacts . The N⋯H/H⋯N contacts contribute only 1.3%.The Hirshfeld surface for pe Fig. 4b and cues Fig. 4c do notet al., 2013I suitable for X-ray diffraction were obtained by recrystallization of the crude product from ethanol solution.The title compound was prepared according to our previously reported method  global, I. DOI: 10.1107/S2056989021002863/hb7963Isup2.hklStructure factors: contains datablock(s) I. DOI: 10.1107/S2056989021002863/hb7963sup3.pdfGeometrical data for title compound and related phases. DOI: Click here for additional data file.10.1107/S2056989021002863/hb7963Isup5.cmlSupporting information file. DOI: 2071135CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


The crystal structure of 1 consists of two crystallographically independent TPRS-{NdIIIO9}  units that form an edge-sharing dinuclear cluster interconnected to neighbouring dimers by the μ3-SO42– anions, yielding a cationic two-dimensional {Nd2(H2O)6(SO4)2}nn+2 sheet. Adjacent cationic layers are then linked via the μ4-glutarato2– ligands into a three-dimensional coordination network. Strong O—H⋯O hydrogen bonds are the predominant interaction in the crystal structure.A three-dimensional coordination polymer, poly[hexaaqua(μ These properties are cooperatively provided by both the inorganic building units and the organic counterparts (Furukawa 2(H2O)6(glutarato)(SO4)2]n (1). The crystal structure reveals that the glutarates act as bridging ligands binding the cationic {Nd2(H2O)6(SO4)2}nn+2 sheets into a three-dimensional network.Herein, we report a microwave synthesis of a new three-dimensional coordination polymer, n crystallizes in the monoclinic P21/c space group. There are two crystallographically independent NdIII cations (Nd1 and Nd2), two sulfate anions, and six coordinated water molecules in the asymmetric unit, as illustrated in Fig. 1The coordination network III cations are nine-coordinated to O atoms from one bridging glutarate2−, two chelating glutarate2−, two chelating sulfate anions and three coordinated H2O, adopting a distorted tricapped trigonal–prismatic geometry, TPRS-{NdIIIO9} , forming an edge-sharing dinuclear unit with its symmetry-related NdIIIO9 polyhedron. The NdIII—O bond distances are in the range of 2.383 (2)–2.785 (2) Å, which are reasonable and comparable to those reported for other NdIII coordination polymers such as [Nd(H2O)4(glutarato)]Cl 4(glutarato)]Cl·2H2O 2(glutarato)]·2H2O 4]Cl 4]Cl·2H2O x(glutarato)}nn+  subunits compensated by uncoordinated chloride anions, each of the tetrahedral SO42– ligands in 1 links three adjacent NdIII atoms, forming a neutral two-dimensional network of [Nd2(H2O)6(glutarato)(SO4)2]n. The S—O bond distances are in the range 1.449 (3)–1.485 (2) Å, with O—S—O angles ranging from 107.78 (16) to 111.67 (15)°. The flexible glutarate linker exhibits a .Both Nd1 can be described as a three-dimensional non-porous framework, which is constructed from edge-sharing TPRS-{NdIIIO9} polyhedra linked through sulfate anions, acting as tritopic inorganic linkers, into a cationic [Nd2(H2O)6(SO4)2]nn+2 sheets parallel to the (011) layers, as illustrated in Fig. 3a. It is noteworthy that these sheets also contain large inorganic [Nd(SO4)]4 rings further stabilized by O—H⋯O hydrogen bonds between the water molecules and sulfate anions (Table 1b). This three-dimensional arrangement also features O—H⋯O hydrogen bonds between two water molecules or between a water molecule and oxygen atoms of the glutarate ligands . In total, all but one hydrogen atom from the six crystallographically independent water molecules are involved in hydrogen bonding tetraaquaneodymium chloride] tetraaquadineodymium chloride dihydrate] diaquadineodymium(III) tetrahydrate] diaquadineodymium(III) dihydrate] 3·8H2O , glutaric acid , and 4,4′-bipyridine  in 40.0 mL of deionized water under ambient conditions. The solution was transferred into an open glass reactor and then irradiated by microwaves (800 W) for 10 minutes. The solution was let to cool to ambient temperature. Pale-purple block-shaped crystals crystallized from the solution within a few minutes. FT–IR  of 1: stretchν(O—H) 3364, stretchν(C—H) 2990, asν(COO−) 1531, sν(COO−) 1430, δ(O—H) 1355, sν(S—O) 1101, sν(S—O) 1077, sν(SO42–) 596.Complex Uiso(H) = 1.2Ueq(C). The H atoms from the water molecules were located in the residual electron-density map, and where necessary, refined with distance and angle restraints or riding on the parent oxygen atom.Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989022000159/jq2009sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022000159/jq2009Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022000159/jq2009Isup3.molSupporting information file. DOI: 2107848CCDC reference:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


Pistacia vera is one of the most important cash crops in Iran that is scattered in arid and semi-arid regions. During a survey of plant-parasitic nematodes of pistachio in Ardakan city in Yazd Province, a species of root-lesion nematode was isolated and identified by morphological, morphometrical, and molecular methods as Pratylenchus oleae Palomares-Rius et al., 2014. This species was isolated from several pistachio trees rhizosphere regarding to molecular analysis, D2–D3 expansion segments of 28S rRNA was amplified by PCR and sequenced. The sequence was deposited in GenBank (Accession No. MW338666). Along with the related phylogenetic analysis, placed this species in a monophyletic clade with other Pratylenchus oleae isolates, based on Bayesian inference (BI) phylogeny. A PCR reaction with the P. oleae specific primer set produced a 547 bp fragment. This is the first report of P. oleae infecting Pistachio tree in the world.Pistachio,  Pistacia. P. vera is one of the most important commercial cultivars among the 11 identified species. More than 498,000 hectares of Iranian lands are cultivated with pistachio trees and its product with nearly 173000 tones has a high domestic and foreign acceptance  and crushed with a microhomogeniser Vibro Mixer . Two microliters proteinase K (600 μg/ml)  were added and the tubes were frozen at ‒80°C for at least 10  min and then incubated at 65°C (1 hr) and 95°C (10  min) consecutively. After incubation, the tubes were centrifuged for 2  min at 14,000  rpm and kept at ‒20°C until use. The PCR was carried out in a 30  μl reaction comprising of 2  µL DNA template, 1  μl forward and reverse primers, 15  μl Taq DNA Polymerase 2 × Master Mix (Ampliqon), and 11  μl distilled water. The forward primer D2A (5′-ACAAGTACCGTGA GGGAAAGTTG-3′) and the reverse primer D3B (5′-TCGGAAGGAACCAGCTACTA-3′) (P. oleae-specific primer pairs Poleae_fw1_4_36 (5′-GACAGATTAGAATGGAATCTGTTCG-3′) and Poleae_rv1_ _525_551 (5′ATCGCTTTTGGATTCAATAATATA-3′) as described by P. oleae sequence was aligned with related sequences from GenBank through National Center for Biotechnology Information (NCBI) BLASTn homology search, using ClustalW implemented by MEGA version 10.0. The GTR + I + G model was selected as the best by jModeltest v.2.1.10. Bayesian tree generated using the Bayesian inference method as implemented in the program MrBayes v.3.2.6   were use v.3.2.6 . Markov datasets .P. oleae, found in roots and around soil from Pistachio tree in Iran are presented in P. oleae as well. P. oleae is characterized by morphological features in females: Vermiform body, lip region with three annuli, slightly offset. Stylet well developed with distinct rounded knobs slightly directed anteriorly. Lateral field with four incisures, metacorpus oval to rounded, isthmus rather short, surrounded by nerve ring, pharyngeal glands well developed, with rather long ventral overlap. Excretory pore either slightly anterior pharyngo-intestinal junction or opposite. Genital branch with one row of oocyte, spermathecal rounded, vagina a straight tube. Vulva transverse slit. Post-vulval uterine sac short, not differentiated. Tail sub-cylindrical, terminus rounded to conical, smooth, male unknown : L  =  522.5 ± 34.9 (463.0–565.0) μm; a  =  28.3 ± 1.6 (25.7–31.0)  μm; b  =  5.9 ± 0.4 (5.2–6.7)  μm; c  =  20.2 ± 0.8 (19.1–21.0) μm; Stylet length  =  17.3 ± 0.7 (16.0–18.0) μm; Median bulb 55.6 ± 2.3 (51.0–59.0) μm; Anterior to Excretory Pore 88.9 ± 2.7 (85.0–93.0) μm; Anterior end to pharyngeal junction 89.0 ± 4.6 (78.0–94.0) μm; Pharynx Length 123.5 ± 3.4 (118.0–129.0) μm; Max. Body diam. 18.5 ± 1.0 (17.0–20.0) μm; Body diam. at anus 12.6 ± 0.5 (12.0–13.0)  μm and tail length  =  25.9 ± 1.7  (22.0–27.0)  μm   μm .P. oleae belong to Spain  and Tunisia . Pratylenchus capsici, 9 isolates are most similar to P. oleae, but differs in presence of males, a functional spermatheca, a larger body and shorter stylet  with P. oleae Spain isolate (KJ510861) and 99% (2–7 nucleotide differences) similar to P. oleae Spain isolates  and P. oleae Tunisia isolates , while P. dunensis and P. penetrans are 89% and 87% similar (61 and 82 nucleotide differences with 6 and 13 Gaps). The gap variations and nucleotide differences between 9 isolates of P. capsici and P. oleae (MW338666) are between 3–9 and 30–37 nucleotides respectively  and one location of Tunisia (Ouled Chamekh)  , But somP. oleae deposited in GenBank database. The test population had no gap (base pair), which was different to those of P. oleae from Spain (KJ510855 and KJ510861) with 100% similarity, one gap with Spain (KJ510856 and KJ510857) and Tunisia (KJ510858) with 99% similarity respectively and two gaps with Tunisia (KJ510859 and KJ510860) with 99% similarity. In summary, Iranian P. oleae was isolated from pistachio trees in Ardakan, Yazd province. It is reported for the first time from Iran and pistachio tree in the world. This nematode can be considered as a risk to the economy in pistachio orchards.The sequence of Iranian isolate showed high nucleotide similarity with D2-D3 region of


The Mg atoms in the title compound are located at an interstitial site of the Dy2.1B37Si9-type structure with an occupancy of 0.5. The (001) layers of B12 icosahedra stack along the c-axis direction with shifting in the  direction. A three-dimensional framework structure of the layers is formed via B—Si bonds and {Si8} units of [Si]3—Si—Si—[Si]3.Single crystals of a novel sodium–magnesium boride silicide, Na The structure is composed of B12 icosahedral clusters: the B—B distances of the 30 distinct bonds in the cluster are in the range of 1.791 (3)–1.843 (5) Å and the average distance is 1.811 Å  Å] on the (001) plane and form layers that stack along the c axis with a sequence of ABCABC by shifts of   Å for Si3—Si3 and 2.3951 (9) Å for Si2—Si3 are comparable with the bond length in crystalline silicon (2.35 Å). The bond angles of Si2—Si3—Si2 and Si2—Si3—Si3 are 113.86 (3)° and 104.61 (4)°, respectively, which are distorted from the regular tetrahedral bond angle of 109.47°. The Si2—B1 distance is 2.043 (2) Å, which is close to the Si—B distances (1.973–2.027 Å) found in β-silicon boride, SiB3     Å] and Si1/B5—Si1/B5 pairs that bind to the B atoms at B3 connect the B12 layers of Na3MgB37Si9 .The framework structure of Bi9 Fig. 1. Because8} unit between the B12 cluster layers. The Na1—Si2 distance is 2.8620 (4) Å and the Na1—B1 and Na1—B2 distances are 2.811 (2) and 2.793 (2) Å, respectively. These distances are almost the same as the Na—Si distance of Na4Si4 [2.878 (3) Å; Morito et al., 201515  Å and 2.333 (3) Å, respectively, which are close to the Mg—Si (2.436 Å) and Mg—B distances (2.353 Å) in MgB12Si2     provide six and 6.3 electrons, respectively, and approximately six electrons are supplied from RE in REx1–B12Siδ3.3–   and REx1–B36Si9C . The lattice constants and unit-cell volume of Mg3B36Si9C are a = 10.0793 Å, c = 16.372 Å, and V = 1440.4 Å3   are a = 10.046–10.095 Å, c = 16.298–16.467 Å, and V = 1429–1454 Å3  are a = 10.000–10.096 Å, c = 16.225–16.454 Å, and V = 1405–1452 Å3    . Then, 10 mg of B2O3 powder  were added to the crucible, which was stacked on another BN crucible containing 30 mg of Mg powder , and these crucibles were encapsulated in a stainless steel container  with Ar gas. The container was heated at 1373 K for 24 h using an electric furnace. After cooling, the crucible was taken out from the reaction container, and any Na and NaSi remaining in the crucible were reacted and removed with 2-propanol and ethanol. Then, the sample was washed with pure water to remove water-soluble compounds such as sodium borate and alkoxide produced by the reaction of Na and alcohol to leave black plates of the title compound. An electron probe microanalyzer  was used to analyze the composition of the obtained single crystal as Na 5.49 (8), Mg 2.37 (7), B 74.8 (7), Si 17.3 (4) atom %, which is nearly matched by Na3MgB37Si9 . Other elements such as O were not found.Na metal pieces , crystalline B powder  and Si powder  were weighed in a BN crucible , with a molar ratio of Na:B:Si = 5:4:3  in a high-purity Ar-filled glove box  twinning, which reduced the R-value  from 0.0651 to 0.0380.Crystal data, data collection and structure refinement details are summarized in Table 310.1107/S2056989022000494/hb8005sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989022000494/hb8005Isup2.hklStructure factors: contains datablock(s) I. DOI: 2141726CCDC reference:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


S(6) ring motif. Intermolecular N—H⋯N and C—H⋯N hydrogen bonds, as well as N—H⋯π and C—H⋯π interactions create a three-dimensional network in the crystal.The molecular conformation of the title compound is stabilized by an intramolecular O—H⋯O hydrogen bond, forming an  20H15Cl2N3O2, is stabilized by an intramolecular O—H⋯O hydrogen bond, forming an S(6) ring motif. The central pyridine ring is almost planar [maximum deviation = 0.074 (3) Å]. It subtends dihedral angles of 86.10 (15) and 87.17 (14)°, respectively, with the phenyl and dichlorophenyl rings, which are at an angle of 21.28 (15)° to each other. The =C(—OH)CH3 group is coplanar. In the crystal, molecules are linked by intermolecular N—H⋯N and C—H⋯N hydrogen bonds, and N—H⋯π and C—H⋯π interactions, forming a three-dimensional network. The most important contributions to the crystal packing are from H⋯H (33.1%), C⋯H/H⋯C (22.5%), Cl⋯H/H⋯Cl (14.1%), O⋯H/H⋯O (11.9%) and N⋯H/H⋯N (9.7%) interactions.The molecular conformation of the title compound, C A strong intramolecular O2—H2⋯O1 hydrogen bond  ring motif  create a three-dimensional network in the crystal  in the range of −0.4290 to 1.5192 a.u. The red patches that appear around N2 are caused by the intermolecular N3—H3A⋯N2 and C16—H16⋯N2 interactions, which are important in the packing of the title molecule. Bright red dots near N2 and amine hydrogen atoms H3A and H3B highlight their functions as hydrogen-bonding acceptors and donors, respectively; these also appear as blue and red areas on the Hirshfeld surface mapped over electrostatic potential , C⋯H/H⋯C , Cl⋯H/H⋯Cl , O⋯H/H⋯O  and N⋯H/H⋯N . Other Cl⋯C/C⋯Cl, C⋯C, Cl⋯O/O⋯Cl, Cl⋯N/N⋯Cl, N⋯C/C⋯N, O⋯N/N⋯O, Cl⋯Cl, O⋯C/C⋯O and N⋯N contacts contribute less than 2.1% to Hirshfeld surface mapping and have little directional influence on molecular packing  and SETWUK (space group: P21/n), adopt nearly planar structures. The crystal structure of SETWOE is stabilized by intermolecular C—H⋯O and C—H⋯π interactions. The C—H⋯O hydrogen bonds generate rings with 22R(14) and 22R(20) motifs. The crystal structure of SETWUK is stabilized by intermolecular C—H⋯F and C—H⋯π interactions. The C—H⋯F bond generates a linear chain with a C(14) motif. In addition, in SETWOE and SETWUK, intramolecular O—H⋯O interactions are found, which generate an S(6) graph-set motif. No significant aryl–aryl or π–π interactions exist in these structures. All this bears some resemblance to the title compound.The polysubstituted pyridines, SETWOE (space group: PP22R(8) ring motif. In the hydrated salt JEBREQ, the presence of the water molecule prevents the formation of the familiar i.e. R32(8)] is formed involving the sulfonate group, the pyrimidinium cation and the water molecule. Both salts form a supramolecular homosynthon . Their isotropic displacement parameters were refined using a riding model with Uiso(H) set to either 1.2Ueq(N) for the NH2 group or 1.5Ueq(O) for the OH group. The C-bound H atoms were positioned geometrically (C—H = 0.95–1.00 Å) and allowed to ride on their parent atoms, with Uiso(H) = 1.5Ueq(C) for the methyl group and Uiso(H) = 1.2Ueq(C) for aromatic and methine H atoms.Crystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989021007994/yz2010sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021007994/yz2010Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021007994/yz2010Isup3.cmlSupporting information file. DOI: 2101203CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


R/S,E)-2-(4-hydroxyphenyl)-4-(2-phenylhydrazin-1-ylidene)chromane-5,7-diol ethanol monosolvate, has been determined. The packing is assisted by C—H⋯O, O—H⋯O, O—H⋯N and O—H⋯C(π) type hydrogen bonds, and the pyran ring has an envelope pucker.The structure of racemic -2-(4-hydroxyphenyl)-4-(2-phenylhydrazin-1-ylidene)chromane-5,7-diol ethanol monosolvate, C21H18N2O4·C2H6O, in a centrosymmetric lattice is reported. The two racemates occupy the same position in the asymmetric unit – a disordered mixed enantiomeric structure. Hydrogen bonds of the type O—H⋯C(π) in addition to typical C—H⋯O, O—H⋯O and O—H⋯N are identified. A positional disorder is seen in the solvent molecule (ethanol) as well. The phenylhydrazone group is nearly coplanar with the chromane ring system [dihedral angle = 15.5 (1)°], while the the 4-hydroxyphenyl ring is perpendicular [dihedral angle = 87.2 (1)°] to the chromane. The pyran ring has an envelope pucker .The crystal structure of racemic ( Enantiomeric structures in centrosymmetric lattices have been discussed by Flack (2003Q = 0.363 (3) Å, θ = 57.6 (3)°; and for the enantiomer: Q = 0.364 (3) Å, θ = 127.4 (4)°]. An intramolecular O—H⋯N hydrogen bond exists between one of the hydroxy groups on the chromane ring and the nitrogen of the hydrazone group  molecule in 1:1 ratio, yielded a disordered mixed enantiomeric crystal in a centrosymmetric lattice  type hydrogen-bond interactions between the solvent ethanol and phenyl ring are observed Table 1. The phe al. 1993. The strce Fig. 2.et al., 2016Chemical context section. The most similar structures for which crystal data have been reported include acyl hydrazone derivatives of 2-phenylchroman-4-one and hesperetin. In particular, crystal structures for 2′-[2-(4-fluorophenyl)chroman-4-ylidene]isonicotinohydrazide -amino}benzamide  was performed in the Cambridge Structural Database , 0.97 Å (CH2), 0.86 Å (NH) or 0.82 Å (OH). Isotropic displacement parameters for these atoms were set to 1.2  or 1.5  times Ueq of the parent atom. Idealized Me of the ethanol molecule were refined as rotating group(s): C22A and C22B (H22A through F) and its idealized tetrahedral OH refined as a rotating group: O5A and O5B .Crystal data, data collection and structure refinement details are summarized in Table 210.1107/S2056989022002079/jy2016sup1.cifCrystal structure: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989022002079/jy2016Isup3.molSupporting information file. DOI: Click here for additional data file.10.1107/S2056989022002079/jy2016Isup3.cmlSupporting information file. DOI: 2153764CCDC reference:  crystallographicinformation; 3D view; checkCIF reportAdditional supporting information:


Z-shaped conformation with the carboxyl group nearly coplanar with the dihydroquinoline unit. In the crystal, two sets of C—H⋯O hydrogen bonds form chains along the b-axis direction, which are connected into corrugated layers parallel to (103) by additional C—H⋯O hydrogen bonds. The layers are connected by C—H⋯π(ring) interactions.The molecule adopts a  20H15NO3, adopts a Z-shaped conformation with the carboxyl group nearly coplanar with the dihydroquinoline unit. In the crystal, corrugated layers are formed by C—H⋯O hydrogen bonds and are stacked by C—H⋯π(ring) interactions. Hirshfeld surface analysis indicates that the most important contributions to the crystal packing are from H⋯H (43.3%), H⋯C/C⋯H (26.6%) and H⋯O/O⋯H (16.3%) interactions. The optimized structure calculated using density functional theory at the B3LYP/ 6–311 G level is compared with the experimentally determined structure in the solid state. The calculated HOMO–LUMO energy gap is 4.0319 eV.The title molecule, C This moiety is planar to within 0.0340 (6) Å (r.m.s. deviation = 0.0164) with N1 and C9 being, respectively, 0.0340 (6) and −0.0279 (7) Å from the mean plane, resulting in a slight twist at this location. The carboxyl group is nearly coplanar with the dihydroquinoline as seen from the 1.04 (5)° dihedral angle between the plane defined by C7/C13/O2/O3 and that of the dihydroquinoline (C1–C9/N1/O1). This is likely due, in part, to the intramolecular C5—H5⋯O2 interaction Table 1. The prob-axis direction, which are connected by inversion-related pairs of C4—H4⋯O2 hydrogen bonds . The positive electrostatic potential (blue region) over the surface indicates hydrogen-donor potential, whereas the hydrogen-bond acceptors are represented by negative electrostatic potential (red region).The nd Fig. 4b. The pet al., 2007a, while those delineated into H⋯H, H⋯C/C⋯H, H⋯O/O⋯H, C⋯C, O⋯C/C⋯O, H⋯N/N⋯H, N⋯C/C⋯N and N⋯O/O⋯N contacts are illustrated in Fig. 5b–i, respectively, together with their relative contributions to the Hirshfeld surface (HS). The most important interaction is H⋯H, contributing 43.3% to the overall crystal packing, which is reflected in Fig. 5b as widely scattered points of high density due to the large hydrogen content of the molecule, with its tip at de = di = 1.19 Å. In the presence of C—H interactions, the pair of characteristic wings in the fingerprint plot delineated into H⋯C/C⋯H contacts  has tips at de + di = 3.07 Å. The pair of scattered points of spikes in the fingerprint plot delineated into H⋯O/O⋯H contacts  have tips at de + di = 2.08 Å. The C⋯C contacts  have tips at de + di = 3.34 Å. The O⋯C/C⋯O contacts, Fig. 5f, contribute 1.5% to the HS and appear as a pair of scattered points of spikes with tips at de + di = 3.55 Å. The H⋯N/N⋯H contacts  have tips at de + di = 3.28 Å. Finally, the C⋯N/N⋯C and O⋯N/N⋯ O contacts, Fig. 5h–i, contribute only 0.5% and 0.1% respectively to the HS and have a low-density distribution of points.The overall two-dimensional fingerprint plot , hardness (η), ionization potential (I), dipole moment (μ), electron affinity (A), electrophilicity (ω) and softness (σ) are collated in Table 3ΔE = ELUMO − EHOMO) of the molecule is 4.0319 eV, and the frontier molecular orbital energies, EHOMO and ELUMO, are −6.3166 and −2.2847 eV, respectively.The structure in the gas phase of the title compound was optimized by means of density functional theory. The density functional theory calculation was performed by the hybrid B3LYP method and the 6–311 G basis-set, which is based on Becke’s model , benzyl chloride  and tetra n-butylammonium bromide as a catalyst in DMF (30 mL) was stirred at room temperature for 48 h. After removal of the salts by filtration, the solvent was evaporated under reduced pressure and the residue obtained was dissolved in dichloromethane. The organic phase was dried over Na2SO4 and concentrated under vacuum. The crude product obtained was purified by chromatography on a column of silica gel (eluent: hexane/ ethyl acetate: 9/1). 1H NMR  δ ppm: 3.08 ; 4.37 ; 5.12 ; 7.08–8.74 ; 13C NMR  δ ppm: 34.3 (CH3—N); 66.2 (CH2—O); 72.1 (–C≡); 73.2 (CH≡); 115.6-148.7 (CHarom and Cquat arom); 162. 5 (C=Oquinol); 168.2 (C=Ocarboxyl). MS (ESI): m/z = 318 (M + H)+.A mixture of 2-oxo-1-(prop-2-yn-1-yl)-1,2-dihydroquinoline-4-carboxylic acid , KCrystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989021007416/tx2040sup1.cifCrystal structure: contains datablock(s) global, I. DOI: 10.1107/S2056989021007416/tx2040Isup3.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021007416/tx2040Isup3.cmlSupporting information file. DOI: 2097267CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Scientific Reports 10.1038/s41598-021-02868-x, published online 10 December 2021Correction to: The original version of this Article contained an error in the legend of Figure 1.a) Garadagh mud volcano in onshore Azerbaijan (40.24°N and 49.51°E).”“(now reads:(a) Otmanbozdag mud volcano (within the Garadagh structure) in onshore Azerbaijan (40.24°N and 49.51°E).”“The original Article has been corrected.


This compound crystallized in the space group Pη5-coordinated Cp′ rings and a κ3-coordinated (BH4)− ligand.New syntheses have been developed for the synthesis of (borohydrido-κ For example, κ1, κ2, and κ3 U–(BH4) binding has previously been reported 4  (Cp′ = C5H4SiMe3), made in 1992 − within the tris-cyclopentadienyl uranium platform using single-crystal X-ray diffraction. Toward this end, we developed new synthetic routes to the Cp′3U(BH4) compound.Actinide borohydrides have been of interest since the 1940s, owing to their potential volatility and applied use in vapor deposition technologies for the production of thin films  compound was originally synthesized by reacting Cp′3UH with H3B-PPh3  from Cp′3UH with H3B-PPh3 in hot THF solvent. We also observed Cp′3U(BH4) could be prepared in high yield (96%) by reacting Cp′3UI with NaBH4 in the presence of 15-crown-5. When this reaction was carried out in toluene at room temperature, the I− ligand was substituted by the (BH4)− anion. Another method we developed for synthesizing Cp′3U(BH4) involved reacting U(BH4)4 with KCp′ (3 equiv.) in diethyl ether. This reaction, where (BH4)− was substituted by (Cp′)−, also proceeded in high yield (89%). X-ray quality crystals of Cp′3U(BH4) formed at 253 K overnight from diethyl ether solutions.The Cp′3U(BH4) from Cp′3UI over U(BH4)4 because the U(BH4)4 starting material was more challenging to isolate in a chemically pure form. Another interesting comparison between the two synthetic methods involved the substitution chemistry. The (Cp′)− anion displaced (BH4)− from U(BH4)4 and (BH4)− displaced I− in Cp′3UI. Hence, we qualitatively concluded that the stability of the U—X bond for molecular compounds dissolved in organic solvents was largest for (Cp′)−, intermediate for (BH4)−, and lowest for I−. The generality of this conclusion is limited, and we acknowledge the solubility of the other reaction products (such as NaI) might significantly influence the substitution chemistry on uranium.Of our two synthetic routes, we preferred making Cp′3U(BH4) were refined in the triclinic Pet al., 20104)3(DME)]2(μ-O) , 2.574 (6), 2.584 (6), and 2.635 (7) Å 3U(BH4) was reported to be 2.48 Å 3U(BH4) in the gas phase and in solution predicted U—B distances of 2.533 and 2.557 Å 2U(BH4)2 structures showed similar U—B distances of 2.56 (1) Å for [C5H3(SiMe3)2]2U(BH4)2 2U(BH4)2 2U(BH4)2  range from 2.458–2.500 Å and average 2.48 (2) Å (uncertainty reported as the standard deviation from the mean at 1σ). These uranium–(Cp′ centroid) distances compare well with the 2.473 Å analogous metric in Cp′3UCl  3]  is more acute than the 117.0° angle in Cp′3UCl and other Cp′3UX structures, where the average (Cp′ centroid)—U—(Cp′ centroid) angles were reported as 117 (1)° in Cp′3UI, 112 (2)° in Cp′3U(η1-CH=CH2), and 118.7 (4)° in Cp′3U[Si(SiMe3)3]. The more acute (Cp′ centroid)—U—(Cp′ centroid) angles are complemented by a more obtuse average (Cp′ centroid)—U—B angle of 104.4 (4)° in Cp′3U(BH4), likely due to the close proximity of the (BH4)1− ligand compared with (Cp′ centroid)—U—X angles of 100.0° in Cp′3UCl, 100 (2)° in Cp′3UI, 98 (3)° in Cp′3U(η1-CH=CH2), and 96.7 (9)° in Cp′3U[Si(SiMe3)3], see Table 1The uranium–(Cp′ centroid) distances in Cp′3U(BH4) structure is that all three of the trimethylsilyl groups are oriented in a single direction towards the (BH4)− unit. This orientation has not been observed in other Cp′3U(anion) and Cp′3U(μ-dianion)UCp′3 structures, which are shown in Figs. 23UCl structure  3] complex 3]1− unit. Since Cp′3U(BH4) has the smallest mono-anion of the Cp′3U(anion) complexes and the correspondingly smallest (Cp′ centroid)—U—(Cp′ centroid), and the largest (Cp′ centroid)—U—X angles, the orientation of the silyl groups could occur due to steric factors. However, it is also possible that some dispersion forces between the (BH4)− and the trimethylsilyl groups could contribute to the orientation  piano-stool complexes: (C5H5)U(BH4)3 U(BH4)(SPSMe) Ph2)-1-Me, a λ4-phosphinine with two lateral phosphinosulfide groups, and the tetramethylphosphol (PC4Me4) compound (PC4Me4)(C8H8)U(BH4)(THF) 2, where Ring = C5H5 2  complex [LU(BH4)][B(C6F5)4] was also in the database  borohydrides, such as the mono borohydride [(PC4Me4)2U(BH4)]2 U(BH4)3]2 2(μ-X) where X = O2− 2− 4(μ-L)  complexes with Cp′3U(BH4) compound was also characterized by 1H, 11B{1H}, 13C{1H}, and 29Si{1H} multi-nuclear NMR spectroscopy. It was of particular interest to examine the 29Si{1H} spectrum for comparison with previous studies of silicon-containing paramagnetic uranium complexes  and paramagnetically shifted over a range of δ 9.6 to −22.6 ppm, in the 1H NMR spectrum, and a 29Si{1H} resonance at δ −57.4 ppm was observed, typical of other tetravalent uranium complexes − unit showed considerably more shifting and broadening, resonating at δ −59.5 (ν1/2 = 300 Hz) and 79.6 (ν1/2 = 240 Hz) in the 1H and 11B{1H} spectra, respectively. Since the (BH4)− ligand exhibited a single 1H NMR resonance whereas two distinct hydride environments are present in the solid state, it appears that the complex is fluxional in solution. This is in line with previous studies  were sparged with UHP argon (Praxair) and dried by passage through columns containing a copper(II) oxide oxygen scavenger (Q-5) and molecular sieves prior to use or stirred over sodium benzophenone ketyl, briefly exposed to vacuum several times to degas and distilled under vacuum. All ethereal solvents were stored over activated 4 Å molecular sieves. Deuterated solvents (Cambridge Isotopes) used for nuclear magnetic resonance (NMR) spectroscopy were dried over sodium benzophenone ketyl, degassed by three freeze-pump-thaw cycles, and distilled under vacuum before use. The 1H, 11B{1H}, 13C{1H} and 29Si{1H} NMR spectra were recorded on a GN 500, Cryo 500 or Bruker Avance 600 spectrometer operating at 500.2 MHz, 160.1 MHz, 125.8 MHz, and 99.1 MHz for the 500 MHz spectrometers, respectively, and 600.1 MHz, 192.6 MHz, 150.9 MHz and 119.2 MHz for the 600 MHz spectrometer, respectively, at 298 K unless otherwise stated. The 1H and 13C{1H} NMR spectra were referenced internally to solvent resonances, 11B and 29Si{1H} NMR spectra were referenced externally to BF3(Et2O) and SiMe4, respectively, the 29Si{1H} spectra were acquired using the INEPT pulse sequence. The 15-crown-5 (Aldrich) reagent was dried over activated molecular sieves and degassed by three freeze–pump–thaw cycles before use. The NaBH4 (Aldrich) reagent was placed under vacuum (10 −3 Torr) for 12 h before use. The following compounds were prepared following literature procedures: KCp′ 4  was added to a C7D8  solution of Cp′3UI  in a J-Young NMR tube, an excess of 15-crown-5 (1 drop) was added and the tube was sealed and removed from the glovebox and vortexed (30 s). The NaBH4 was not fully soluble in C7D8 even in the presence of 15-crown-5. After 18 h, NMR spectroscopy showed complete conversion to Cp′3U(BH4). The sample was brought back into the glovebox and the volatiles were removed under reduced pressure. The product was then extracted into Et2O, filtered away from white insoluble solids [presumably Na(15-crown-5)I and excess NaBH4] and the volatiles were removed under reduced pressure to give Cp′3U(BH4)  as a wine-red solid. 1H NMR : δ 9.7 , −2.1 , −23.1 , −59.8 , 4H); 11B{1H} NMR : δ 79.1 ; 13C{1H} NMR : δ 233.1 (C5H4SiMe3), 214.0 (C5H4SiMe3), 185.6 (C5H4SiMe3), 0.4 (C5H4SiMe3); 29Si{1H} NMR : δ −57.7 ; 1H NMR : δ 9.6 , −2.0 , −22.6 , −59.3 , 4H); 11B{1H} NMR : δ 79.6 ; 13C{1H} NMR : δ 232.0 (C5H4SiMe3), 214.2 (C5H4SiMe3), 186.5 (C5H4SiMe3), 0.6 (C5H4SiMe3); 29Si{1H} NMR : δ −57.4 .Solid NaBH2O (5 mL) solution of KCp′  was added to a pale-green solution of U(BH4)4 , also dissolved in Et2O (5 mL). White solids precipitated (presumably KBH4) as the solution quickly turned orange and then slowly changed to dark red (30 min). After stirring the mixture for an additional 12 h, volatiles were removed under reduced pressure, and the product was extracted into hexane leaving white solids behind (presumably KBH4). Removal of the volatiles under reduced pressure gave Cp′3U(BH4)  as a dark wine-red solid. X-ray quality crystals were grown from a concentrated ether solution at 253 K.An Ethttps://submission.iucr.org/jtkt/serve/z/Utgd9EjfTrqJVoXA/zz0000/0/.Crystal data, data collection and structure refinement details are summarized in Table 23 moieties, respectively. Methyl torsion angles were not refined but constrained to be staggered. The borohydride H atoms were located from a difference-Fourier map and their positions were freely refined. Uiso(H) values were set to a multiple of Ueq(C/B) with 1.5 for CH3 and BH4 and 1.2 for C—H units, respectively.C—H bond distances were constrained to 0.95 Å for cyclopentadienyl C—H moieties, and to 0.98 Å for aliphatic CH10.1107/S2056989021002425/zl5005sup1.cifCrystal structure: contains datablock(s) I. DOI: 10.1107/S2056989021002425/zl5005Isup3.hklStructure factors: contains datablock(s) I. DOI: 2067867CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information:


Inversion-related chains pack to form layers parallel to the bc plane.The cyclohexene ring of the tetrahydroisoquinoline unit is non-planar. The two 4-chlorophenyl groups extend away from one side of this unit while the hydroxyl and acetyl groups extend away from the opposite side. An intramolecular O—H⋯O hydrogen bond fixes the rotational orientation of the acetyl group. In the crystal, N—H⋯O hydrogen bonds form chains of molecules extending along the  28H25Cl2N3O3S, the heterocyclic portion of the tetrahydroisoquinoline unit is planar while the cyclohexene ring adopts a twist-boat conformation. The two 4-chlorophenyl groups extend away from one side of this unit while the hydroxyl and acetyl groups extend away from the opposite side and form an intramolecular O—H⋯O hydrogen bond. The crystal packing consists of layers parallel to the bc plane. A Hirshfeld surface analysis of the crystal structure indicates that the most important contributions to the crystal packing are from H⋯H (37.3%), Cl⋯H/H⋯Cl (17.6%), O⋯H/H⋯O (11.1%), C⋯H/H⋯C (10.9%) and N⋯H/H⋯N (9.7%) interactions.In the title molecule, C The conformation of this ring approximates a twist-boat conformation. The best planes through the C10–C15 and C23–C28 rings are inclined to the N1/C5–C9 plane by 76.05 (6) and 74.04 (6)°, respectively. The acetyl group on C2 is in an equatorial position while the hydroxyl group on C3 is axial and these are syn to one another. The C10–C15 ring attached to C1 is close to equatorial and anti with respect to both other substituents  Å, O3⋯Cg1ii = 3.6287 (11) Å and C22—O3⋯Cg1ii = 115.38 (8)°; symmetry code: (ii) x, y, z; where Cg1 is the centroid of the N1/C5-C9 ring) are also observed in the crystal structure.In the crystal, helical chains extending along the s Table 1. Inversis Table 1. In addiCrystal Explorer 17.5 , Cl⋯H/H⋯Cl , O⋯H/H⋯O , C⋯H/H⋯C  and N⋯H/H⋯N . The other contacts are negligible with individual contributions of less than 2.9% and are given in Table 3Fig. 5et al., 2016et al., 2017et al., 2020H-spiroisoquinolin]-2(1H)-one -yl]-1,2-diphenylethan-1-ol -6,7-dimethoxy-3-(methoxydiphenylmethyl)-1-phenyl-1,2,3,4-tetrahydroisoquinoline  and graph-set motifs of the hydrogen-bonding network. In the crystal of DUSVIZ, molecules are linked via C—H⋯O hydrogen bonds. For the major disorder component, these form C(11) chains that propagate parallel to the a axis. In the crystal of AKIVUO, a layer structure with the layers parallel to  and (10via weak C—H⋯O and C—H⋯π(ring) interactions. In the crystal structure of ENOCIU, various C—H⋯π and C—H⋯O interactions link the molecules. In the crystal of NIWPAL, the molecules are linked by N—H⋯O intermolecular hydrogen bonds involving the sulfonamide function to form an infinite two-dimensional network parallel to the (001) plane.In the crystal of NAQRIJ, dimers form through complementary sets of inversion-related O—H⋯O and C—H⋯O hydrogen bonds. These are connected into zigzag chains along the H)-thione,  with N-(4-chlorophenyl)-2-chloroacetamide  and  of anhydrous sodium acetate in pure ethanol (30 ml) for 1 h as shown in Fig. 6The title compound was obtained by refluxing of 7-acetyl-8-(4-chlorophenyl)-4-cyano-1,6-dimethyl-6-hydroxy-5,6,7,8-tetrahydroisoquinoline-3(2−1 (O—H), 3277 cm−1 (N—H), 2991, 2920 cm−1 , 2217 cm−1 (C≡N), 1694 , 1666 cm−1 . 1H NMR : δ 10.95 ; 8.17–8.24 ; 7.79–7.81 ; 7.26–7.32 ; 7.03–7.05 ; 4.88 ; 4.53–4.55 ; 4.19–4.20 ; 3.24–3.29 ; 2.87–2.90 ; 2.13 ; 1.86 ; 1.27 . Analysis calculated for C28H25Cl2N3O3S (554.47): C 60.65%, H 4.54%, N 7.58%, S 5.78%. Found: C 60.34%, H 4.57%, N 7.68%, S 5.97%.IR: 3522 cmCrystal data, data collection and structure refinement details are summarized in Table 410.1107/S2056989021003674/vm2246sup1.cifCrystal structure: contains datablock(s) I, global. DOI: 10.1107/S2056989021003674/vm2246Isup2.hklStructure factors: contains datablock(s) I. DOI: Click here for additional data file.10.1107/S2056989021003674/vm2246Isup3.cmlSupporting information file. DOI: 2075592CCDC reference:  crystallographic information; 3D view; checkCIF reportAdditional supporting information: