Molecular Insights into Water-Chloride and Water-Water Interactions in the Supramolecular Architecture of Aconine Hydrochloride Dihydrate.

Despite the previous preparation of aconine hydrochloride monohydrate (AHM), accurate determination of the crystal's composition was hindered by severely disordered water molecules within the crystal. In this study, we successfully prepared a new dihydrate form of the aconine hydrochloride [C25H42NO9+Cl-·2(H2O), aconine hydrochloride dihydrate (AHD)] and accurately refined all water molecules within the AHD crystal. Our objective is to elucidate both water-chloride and water-water interactions in the AHD crystal. The crystal structure of AHD was determined at 136 K using X-ray diffraction and a multipolar atom model was constructed by transferring charge-density parameters to explore the topological features of key short contacts. By comparing the crystal structures of dihydrate and monohydrate forms, we have observed that both AHD and AHM exhibit identical aconine cations, except for variations in the number of water molecules present. In the AHD crystal, chloride anions and water molecules serve as pivotal connecting hubs to establish three-dimensional hydrogen bonding networks and one-dimensional hydrogen bonding chain; both water-chloride and water-water interactions assemble supramolecular architectures. The crystal packing of AHD exhibits a complete reversal in the stacking order compared to AHM, thereby emphasizing distinct disparities between them. Hirshfeld surface analysis reveals that H···Cl- and H···O contacts play a significant role in constructing the hydrogen bonding network and chain within these supramolecular architectures. Furthermore, topological analysis and electrostatic interaction energy confirm that both water-chloride and water-water interactions stabilize supramolecular architectures through electrostatic attraction facilitated by H···Cl- and H···O contacts. Importantly, these findings are strongly supported by the existing literature evidence. Consequently, navigating these water-chloride and water-water interactions is imperative for ensuring storage and safe processing of this pharmaceutical compound.

From antioxidant defense to genotoxicity: Deciphering the tissue-specific impact of AgNPs on marine clam Ruditapes philippinarum.

The escalating use of silver nanoparticles (AgNPs) across various sectors for their broad-spectrum antimicrobial capabilities, has raised concern over their potential ecotoxicological effects on aquatic life. This study explores the impact of AgNPs (50 µg/L) on the marine clam Ruditapes philippinarum, with a particular focus on its gills and digestive glands. We adopted an integrated approach that combined in vivo exposure, biochemical assays, and transcriptomic analysis to evaluate the toxicity of AgNPs. The results revealed substantial accumulation of AgNPs in the gills and digestive glands of R. philippinarum, resulting in oxidative stress and DNA damage, with the gills showing more severe oxidative damage. Transcriptomic analysis further highlights an adaptive up-regulation of peroxisome-related genes in the gills responding to AgNP-induxed oxidative stress. Additionally, there was a noteworthy enrichment of differentially expressed genes (DEGs) in key biological processes, including ion binding, NF-kappa B signaling and cytochrome P450-mediated metabolism of xenobiotics. These insights elucidate the toxicological mechanisms of AgNPs to R. philippinarum, emphasizing the gill as a potential sensitive organ for monitoring emerging nanopollutants. Overall, this study significantly advances our understanding of the mechanisms driving nanoparticle-induced stress responses in bivalves and lays the groundwork for future investigations into preventing and treating such pollutants in aquaculture.

Seawater quality criteria and ecotoxicity risk assessment of zinc oxide nanoparticles based on data of resident marine organisms in China.

Water quality criteria (WQC) for zinc oxide nanoparticles (ZnO NPs) are crucial due to their extensive industrial use and potential threats to marine organisms. This study conducted toxicity tests using marine organisms in China, revealing LC50 or EC50 values for ZnO NPs ranging from 0.36 to 95.6 mg/L across seven species, among which the salinity lake crustacean zooplankton Artemia salina exhibited the highest resistance, while diatom Phaeodactylum tricornutum the most sensitive. Additionally, the EC10 or maximum acceptable toxicant concentration (MATC) values for ZnO NPs were determined for five species, ranging from 0.03 to 2.82 mg/L; medaka Oryzias melastigma demonstrated the highest tolerance, while mysis shrimp Neomysis awatschensis the most sensitive. Based on the species sensitivity distribution (SSD) method, the derived short-term and long-term WQC for ZnO NPs were 138 µg/L and 8.37 µg/L, respectively. These values were further validated using the sensitive species green algae Chlorella vulgaris, confirming effective protection. There is no environmental risk observed in Jiaozhou Bay, Yellow River Estuary and Laizhou Bay in the northern coastal seas of China. This study provides important reference data for the establishment of water quality standards for nanoparticles.

Therapeutic effect of demethylated hydroxylated phillygenin derivative on Helicobacter pylori infection.

Modifying and transforming natural antibacterial products is a novel idea for developing new efficacious compounds. Phillygenin has an inhibitory effect on H. pylori. The aim of the present study was to prepare a phillygenin derivative (PHI-Der) through demethylation and hydroxylation. The minimum inhibitory concentration of 18 strains of H. pylori from different sources was 8-32 µg/mL in vitro, and the activity increased 2-8 times than that of phillygenin. PHI-Der could significantly inhibit the colonization of H. pylori in vivo, reduce the inflammatory response, and promote the repair of inflammatory damage. Further, we used SwissTargetPrediction to predict that its main targets are ALOX5, MCL1, and SLC6A4, and find that it can inhibit bacterial biofilm formation and reduce bacterial infection of cells. It can enhance the intracellular oxidative capacity of H. pylori to inhibit H. pylori growth. Further, it could prevent the oxidation of H. pylori-infected cells and reduce the inflammatory response, which plays a role in protection. In conclusion, compared to phillygenin, PHI-Der had better antibacterial activity and was more effective in treating H. pylori infection. It has characteristics of high safety, specificity, resistance to drug resistance and better antibacterial activity than phillygenin, it's a good antioxidant for host cells.

BanXiaXieXin decoction treating gastritis mice with drug-resistant Helicobacter pylori and its mechanism.

BACKGROUND: Helicobacter pylori (H. pylori) is the main pathogen that causes a variety of upper digestive diseases. The drug resistance rate of H. pylori is increasingly higher, and the eradication rate is increasingly lower. The antimicrobial resistance of H. pylori is an urgent global problem. It has been confirmed that Banxia Xiexin decoction (BXXXT) demonstrates the effects of treating gastrointestinal diseases, inhibiting H. pylori and protecting gastric mucosa. The purpose of the present study is to further explore the therapeutic effects of BXXXT on drug-resistant H. pylori. AIM: To confirm that BXXXT demonstrates therapeutical effects in vivo and in vitro on gastritis mice with drug-resistant H. pylori and explain its mechanism to provide an experimental basis for promoting the application of BXXXT. METHODS: The aqueous extract of BXXXT was gained by water decocting method. The inhibitory effect of the aqueous extract on H. pylori was detected by dilution in vitro; drug-resistant H. pylori cells were used to build an acute gastritis model in vivo. Thereafter, the model mice were treated with the aqueous extract of BXXXT. The amount of H. pylori colonization, the repair of gastric mucosal damage, changes of inflammatory factors, apoptosis, etc., were assessed. In terms of mechanism exploration, the main medicinal compositions of BXXXT aqueous extract and the synergistic bacteriostatic effects they had demonstrated were analyzed using mass spectrometry; the immune function of peripheral blood cells such as CD3+ T and CD4+ T of mice with gastritis before and after treatment with BXXXT aqueous extract was detected using a flow cytometry; the H. pylori transcriptome and proteome after treatment with BXXXT aqueous extract were detected. Differently expressed genes were screened and verification was performed thereon with knockout expression. RESULTS: The minimum inhibitory concentration of BXXXT aqueous extract against H. pylori was 256-512 µg/mL. A dose of 28 mg/kg BXXXT aqueous extract treatment produced better therapeutical effects than the standard triple therapy did; the BXXXT aqueous extract have at least 11 ingredients inhibiting H. pylori, including berberine, quercetin, baicalin, luteolin, gallic acid, rosmarinic acid, aloe emodin, etc., of which berberine, aloe emodin, luteolin and gallic acid have a synergistic effect; BXXXT aqueous extract was found to stimulate the expressions of CD3+ T and CD4+ T and increase the number of CD4+ T/CD8+ T in gastritis mice; the detection of transcriptome and proteome, quantitative polymerase chain reaction, Western blotting and knockout verification revealed that the main targets of BXXXT aqueous extract are CFAs related to urea enzymes, and CagA, VacA, etc. CONCLUSION: BXXXT aqueous extract could demonstrate good therapeutic effects on drug-resistance H. pylori in vitro and in vivo and its mechanism comes down to the synergistic or additional antibacterial effects of berberine, emodin and luteolin, the main components of the extract; the extract could activate the immune function and enhance bactericidal effects; BXXXT aqueous extract, with main targets of BXXXT aqueous extract related to urease, virulence factors, etc., could reduce the urease and virulence of H. pylori, weaken its colonization, and reduce its inflammatory damage to the gastric mucosa.

Supramolecular structure, in vivo biological activities and molecular-docking-based potential cardiotoxic exploration of aconine hydrochloride monohydrate as a novel salt form.

Despite the high profile of aconine in WuTou injection, there has been no preparative technology or structural studies of its salt as the pharmaceutical product. The lack of any halide salt forms is surprising as aconine contains a tertiary nitrogen atom. In this work, aconine was prepared from the degradation of aconitine in Aconiti kusnezoffii radix (CaoWu). A green chemistry technique was applied to enrich the lipophilic-poor aconine. Reaction of aconine with hydrochloride acid resulted in protonation of the nitrogen atom and gave a novel salt form (C25H42NO9+·Cl-·H2O; aconine hydrochloride monohydrate, AHM), whose cation in the crystal structure was elucidated based on extensive spectroscopic and X-ray crystallographic analyses. The AHM crystal had a Z' = 3 structure with three independent cation-anion pairs, with profound conformational differences among the aconine cations. The central framework of each aconine cation was compared with that of previously reported aconitine, proving that protonation of the nitrogen atom induced the structure rearrangement. In the crystal of AHM, aconine cations, chloride anions and water molecules interacted through inter-species O-H...Cl and O-H...O hydrogen bonds; this complex hydrogen-bonding network stabilizes the supramolecular structure. The seriously disordered solvent molecules were treated using the PLATON SQUEEZE procedure [Spek (2015). Acta Cryst. C71, 9-18] and their atoms were therefore omitted from the refinement. Bioactivity studies indicated that AHM promoted in vitro proliferative activities of RAW264.7 cells. Molecular docking suggested AHM could target cardiotoxic protein through the hydrogen-bonding interactions. The structural confirmation of AHM offers a rational approach for improving the pharmaceutical technology of WuTou injection.

Optimization of the traditional processing method for precision detoxification of CaoWu through biomimetic linking kinetics and human toxicokinetics of aconitine as toxic target marker.

ETHNOPHARMACOLOGICAL RELEVANCE: CaoWu (Aconiti Kusnezoffii Radix), well known for its high toxicity leading to fatal ventricular arrhythmias, is detoxified by HeZi (Terminalia Chebula Retz) decoction to prepare ZhiCaoWu (Aconiti Kusnezoffii Radix Preparata) as one part of ingredients of NaRu-3 pill which is used for the treatment of rheumatoid arthritis (RA). Aconitine (AC) is a highly toxic alkaloid of CaoWu and it is used as toxic target marker for the quality control (QC) of ZhiCaoWu. In the traditional processing method, the vanish of astringent or spicy feeling in tongue is the important detoxification indicator of ZhiCaoWu. However, how CaoWu is detoxified to ZhiCaoWu and whether the appropriate content of AC in ZhiCaoWu can be efficiently perceived after the empirical detoxification still lack factual basis. AIM OF THE STUDY: The present study aimed to optimize the traditional processing method for precision detoxification of CaoWu through biomimetic linking kinetics and human toxicokinetics (TK) of AC, with a view of providing insights into the changes of toxic target marker. MATERIALS AND METHODS: CaoWu medicinal slices (Mes) and coarse powder (Cop) were processed by blank HeZi decoction through the soaking method for 7 days. High-performance liquid chromatography (HPLC) was used for the analysis of the samples. The acidity of blank HeZi decoction and HeZi processing decoction was directly determined by pH meter. The non-compartment analysis (NCA) was used to have an intuitive appreciation for AC and pH changes in HeZi processing decoction while the compartment model method was used to build the biomimetic linking kinetics model with the covariate. The inter-species scaling of animal TK parameters was conducted to predict human AC TK profiles. The possible uptake ways of AC (rapid-release or extended-release) for humans were attempted to assess the poisoning risk of AC in NaRu-3 pill. Based on the target content of AC in ZhiCaoWu, the biomimetic linking kinetics model was explored to optimize the traditional processing detoxification method of CaoWu. The assays of determining inflammatory cytokines in lipopolysaccharides (LPS)-induced RAW264.7 cells were performed to investigate the inflammatory modulation effects of AC in vitro. RESULTS: ZhiCaoWu was prepared by eliminating redundant AC in CaoWu through the repeatable replacement of HeZi processing decoction in which its acidity (pH) was affected. AC-pH changes in HeZi processing decoction were adequately depicted by a biomimetic linking kinetics model whose predictive power was determined by comparing the predictions of AC in ZhiCaoWu with the reported data. Rapid-release AC at the converted dose of 111.1 and 417.6 µg (0.011 and 0.042% of AC in NaRu-3 pill) reached maximum blood concentrations of 26.1 and 98.1 ng/mL at 0.3 h, in comparison with minimum human lethal concentration (100 ng/mL). Achieving the target content of AC (0.04%) in ZhiCaoWu or AC (0.011%) in NaRu-3 pill to precisely control the poisoning risk, the potential optimized protocols were that the processing time at 0.2-0.8% of AC in CaoWu was 2.0-4.4 days for Cop and 2.7-6.2 days for Mes. Correspondingly, pH values in HeZi processing decoction were 3.95 and 3.77 for Cop and Mes, respectively. Meanwhile, Lipopolysaccharides (LPS)-induced RAW264.7 cells were exposed to 0, 20, and 200 µM of AC for 12 h and AC at 20 µM enhanced the levels of IL-6, IL-10 and TNF-α. CONCLUSIONS: Thus, for the first time, a biomimetic linking kinetics model was built to optimize the traditional detoxification method. Moreover, pH changes could be developed as surrogate endpoint for guiding the processing detoxification of CaoWu. Notably, setting the content limit of AC (0.011%) was very rational to control the poisoning risk of NaRu-3 pill. In addition, it was possible that there existed the more complex mechanisms of AC for inflammatory modulation in vitro.

Optimization of maintenance therapy of Risperidone with CYP2D6 genetic polymorphisms through an extended translational framework-based prediction of target occupancies/clinical outcomes.

Risperidone, one of the second-generation antipsychotics, can efficiently target dopamine D2 and serotonin 5-HT2A receptors. There actually exists significant implication of CYP2D6 genetic polymorphisms on the metabolic kinetics of risperidone, little is known about the extent of CYP2D6 impacting human D2 and 5-HT2A receptor occupancies as well as the clinical efficacy and efficacy in schizophrenia treatment. Here we assessed the influences of CYP2D6 gene polymorphisms on human target occupancies/clinical outcomes and optimized the maintenance therapy of risperidone. A translational framework, previously developed using in vitro and in vivo information in rats, was used as the basis for integrating the effects of CYP2D6 genetic polymorphisms on target occupancies and clinical outcomes. D2 occupancy as a biomarker was related to Positive and Negative Syndrome Scale (PANSS) response and Simpson-Angus Scale (SAS). The population approach was applied to characterize pharmacokinetic and pharmacodynamic (PK/PD) profiles of risperidone. Non-compartment analysis method was performed to calculate the steady state PK/PD parameters of both risperidone and 9-hydroxyrisperidone. The predictive power of this extended translational framework was determined by comparing the predictions of target occupancies and clinical outcomes with the reported human values of risperidone at clinically suggested dosage of 4.0 mg/day. This extended translational framework was adequately used to predict human target occupancies and clinical outcomes. At the steady state, D2 ROs were 75.8%, 79.3% and 86.0% for CYP2D6 poor metabolizer (PM), intermediate metabolizer (IM) and extensive metabolizer (EM), respectively; 5-HT2A ROs were 96.4%, 97.2% and 98.4% for CYP2D6 PM, IM and EM, respectively; PANSS changes from placebo were -5.3, -7.7 and -11.3 for CYP2D6 PM, IM and EM, respectively; SAS changes from placebo were 0.13, 0.15 and 0.18 for CYP2D6 PM, IM and EM, respectively. The predictions of human D2, 5-HT2A RO, PANSS and SAS changes for risperidone with CYP2D6 genetic polymorphisms were well in line with the reported values in clinic. 5.0, 4.0 and 2.5 mg/day were the equivalent dosages of risperidone for CYP2D6 PM, IM and EM, respectively. The optimized maintenance therapy of risperidone was provided through the Three-Step method and the dosage range was 2.5-5.0 mg/day for three CYP2D6 gene groups in the present study. Taken together, our findings demonstrate that this extended translational framework not only differentiates the effects of CYP2D6 genetic polymorphisms on target occupancies and clinical outcomes, but also constitutes a scientific basis to optimize the maintenance therapy of neuropsychiatric patients in clinic.

Utilization of model-based meta-analysis to delineate the net efficacy of taspoglutide from the response of placebo in clinical trials.

The objective of this study was to develop quantitative models to delineate the net efficacy of taspoglutide on fasting plasma glucose (FPG) and glycosylated hemoglobin (HbA1c) from the response of placebo in type 2 diabetes patients, and further find pharmacodynamic potency of taspoglutide and FPG for half of maximum reduction responses of FPG and HbA1c, respectively. Several PD data about taspoglutide treatments for type 2 diabetes patients were digitalized from the published papers related with the clinical development of taspoglutide. The model based meta-analysis (MBMA) studies for FPG and HbA1c were performed with Monolix 4.2 software. The MBMA successfully described the effects of placebo and taspoglutide on pharmacological indexes of FPG and HbA1c through mono and multiple combination therapies in clinical trials. The pharmacodynamic potency (25.3 pmol/l) produced 50% of maximum responses of FPG (-2.39 mmol/l) from the responses of placebo for FPG (-0.371 mmol/l); the response change of FPG (-1.81 mmol/l) affected 50% of maximum response change (-1.74%) for HbA1c from the response of placebo (-0.253%). The leveraging prior knowledge from the longitudinal MBMA will be utilized to guide clinical development of taspoglutide and further support study designs including optimization of dose and duration of therapy.

The efficacy of placebo-adjusted taspoglutide on body weight reduction in clinical trials.

Taspoglutide has elicited a long-lasting glycemic control effect with favorable body weight loss. The objective of this study was to develop a quantitative model to delineate the net efficacy of taspoglutide on body weight (WT) loss from the response of placebo in type 2 diabetes patients, and further find pharmacodynamic potency of taspoglutide for half of maximum reduction response of WT. Several PD data about taspoglutide treatments for type 2 diabetes patients were digitalized from the published papers. The model based metaanalysis (MBMA) study for WT loss was performed with Monolix 4.3 software. The MBMA successfully described the effects of placebo and taspoglutide on the pharmacological index of WT loss in clinical trials. The pharmacodynamic potency (41.7 pmol/l) produced 50% of maximum response of WT (-1.85 kg) from the responses of placebo (-1.33 kg). The longitudinal MBMA could be utilized to quantitatively describe the efficacy of taspoglutide on body weight loss and may lead to a clinical guideline for treatment of type 2 diabetes patients in the future.

[Mongolian medicine cha gan beng ga regulated activity of biomarker PGC-1α].

OBJECTIVE: To investigate the regulation of Cha Gan Beng Ga on the activity of biomarker PGC-1α in vivo and in vitro, and lay the foundation for studying the efficacy result of Cha Gan Beng Ga on xenograft tumor model and extracting active constituents. METHOD: (1) The coarse powder of Cha Gan Beng Ga was extracted with 70% ethanol solution through heating and refluxing, and finally was used to freeze dry powder. (2) 50 mg x kg(-1) of freeze-dried power was orally administrated to KM and C57BL/6J mice once daily, lasting for 5 consecutive days; different concentrations of extracted materials was given to non-small cell lung cells A549. (3) The expression level of PGC-1α mRNA was quantitatively determined in lung tissue of mice and non-small cell lung cells A549. RESULT: The expression levels of PGC-1α in lung tissue of different mice strains had an increasing tendency. Furthermore, the expression levels of PGC-1α in non-small cell lung cells A549 also had an increasing tendency, showing dose and time-dependent relationships. CONCLUSION: Mongolian Medicine Cha Gan Beng Ga could induce the over-expression of PGC-1α mRNA in lung tissue of mice and in non-small cell lung cells A549. The present results will lay foundation for studying the efficacy result of antitumor and active constitutes in future.

[Influence of several excipients on damp-proof performance of pharmaceutical materials of traditional Chinese medicine].

OBJECTIVE: To study the influence of several excipients on damp-proof performance of pharmaceutical materials of traditional Chinese medicine. METHOD: The moisture absorption rate and parameters of hydroscopicity were used as the evaluation index of the damp-proof property of the complex Chinese medicine and preparation 1 and 2. RESULT: The moisture rate of complex Chinese medicine 1 was 62.54%, the critical relative humidity (CRH) was 38%. The moisture rate of complex Chinese medicine 2 was 16.36%, the CRH was 53%. Excipients had different effect on lower the hyproscopic property of complex Chinese medicine 1 and 2. The initial moisture adsorption velocity of excipients of complex Chinese medicine 1 in a ascending order were dextrin < calcium hydrogen phosphate < micro crystalline cellulose < lactose < ethyl cellulose < mannitol < hydroxypropyl methyl cellulose. The moisture adsorption acceleration of excipients in a ascending order were dextrin = calcium hydrogen phosphate = micro crystalline cellulose < mannitol = ethyl cellulose = lactose < hydroxypropyl methyl cellulose. The moisture adsorption rate of excipients in a ascending order were dextrin < lactose < calcium hydrogen phosphate < ethyl cellulose < micro crystalline cellulose < mannitol < hydroxypropyl methyl cellulose. The initial moisture adsorption velocity of excipients of complex Chinese medicine 2 in a ascending order were mannitol < dextrin < calcium hydrogen phosphate < lactose < ethyl cellulose < micro crystalline cellulose < hydroxypropyl methyl cellulose . The moisture adsorption acceleration of excipients in a ascending order were mannitol = dextrin = calcium hydrogen phosphate < lactose < ethyl celluloselactose < micro crystalline cellulose < hydroxypropyl methyl cellulose. The moisture adsorption rate of excipients in a ascending order were mannitol < dextrin < calcium hydrogen phosphate < lactose < ethyl cellulose < micro crystalline cellulose < hydroxypropyl methyl cellulose. CONCLUSION: The choosing of damp-proof excipients of preparation based on the property of the complex traditional Chinese medicine. The study provided experimental evidences for the research and development of the pharmaceutical materials of traditional Chinese medicine.

[Evaluation of transdermal property in vitro of different breviscapine lipid carrier gels].

OBJECTIVE: To prepare four breviscapine lipid carrier gels and general gel, and to study the transdermal promotive effects of them in vitro. METHOD: Improved Franz-type diffusion cell and isolated rat skin were used as transdermal barrier, and the transdermal property was fitted by pharmacokinetics equations. The unit area cumulative permeation quantity of breviscapine was determined by HPLC method. RESULT: The 24 hours' unit area cumulative permeation quantity of SLN gel was the highest, followed in descending order were transfersomal gel, liposomal gel, NLC gel and general gel. The transdermal property in vitro of SLN gel fitted in with Higuchi pharmacokinetics equation. CONCLUSION: The lipid carrier gels had different transdermal promotive effects to breviscapine, SLN gel was the best one as breviscapine transdermal carrier.