Methylephedrine Hydrochloride

The bionic drug-containing cerebrospinal fluid(CSF) of the herb pair Ephedrae Herba-Armeniacae Semen Amarum was prepared and analyzed by experiments on the chemical composition, effects, and correlations, on the basis of which the central active ingredients of the pair and their mechanisms of relieving dyspnea were clarified and the feasibility of the bionic drug-containing CSF method for the neuropharmacological study of traditional Chinese medicine(TCM) in vitro was explored. UPLC-Q-TOF-MS was used to detect and identify the ingredients in the mass spectra of the cerebrospinal fluid containing the herb pair mixed in three ratios. A total of 40 compounds were identified, and 13 differential components were obtained by OPLS-DA. The comparison with the data from the database of the active ingredients of the herb pair for dyspnea alleviation preliminarily identified ephedrine, pseudoephedrine, and methylephedrine as the main differentially active ingredients of the herb pair in the brain. The blood-brain barrier(BBB) model was established by contact co-culture of rat brain microvascular endothelial cells(RBE4) and astrocytes(AC), and the drug-containing CSF was prepared with the BBB model after obtaining drug-containing serum with the herb pair mixed in different ratios. Then, the mass spectra of RBE4 and AC were compared with that of the drug-containing CSF. It was found that the compositional similarity between the bionic drug-containing CSF and the drug-containing CSF was as high as 91.43%, while the content of differential active ingredients in the bionic systems with different mixture ratios was higher than that in the drug-containing CSF. A Transwell chamber was used to construct a PC12 and AC co-culture system, and 6.4×10~(-3) mol·L~(-1) histamine phosphate was used to establish an in vitro cortical neuronal injury model. After the herb pair in a 2∶1 ratio was used to treat different final concentrations of the bionic drug-containing CSF and the drug-containing CSF, biochemical kits were used to measure the content of neurotransmitters, such as nerve growth factor(NGF), substance P(SP), acetylcholine(Ach), 5-hydroxytryptamine(5-HT), and norepinephrine(NA), and their relative standard deviation(RSD) values were calculated. RT-PCR and Western blot were employed to determine the mRNA and protein levels, respectively, of neurokinin-1 receptor(NK-1R), mitogen-activated protein kinase(MAPK), and proto-oncogene C-Fos(C-FOS). The results showed that compared with the normal group, the model group showed increased content of neurotransmitters and up-regulated mRNA and protein levels of NK-1R, MAPK, and C-FOS. Compared with the model group, the drug intervention groups showed reduced content of neurotransmitters and down-regulated mRNA and protein levels of NK-1R, MAPK, and C-FOS. Moreover, the RSD values of neurotransmitter changes in the bionic drug-containing CSF group were generally smaller than those in the drug-containing CSF group. Regression analyses were performed for the content of the active ingredients of the above two drug-containing systems and each effect indicator. The results showed that each effect indicator had higher correlation with the content of the ingredients in the bionic drug-containing CSF than with that in the drug-containing CSF. In conclusion, ephedrine, pseudoephedrine, and methylephedrine may be the main central active ingredients of the herb pair for relieving dyspnea. They exert their dyspnea-relieving effects by inhibiting the expression of NK-1R, MAPK, and C-FOS in the central nervous system and reducing the content of neurotransmitters such as NGF, SP, Ach, 5-HT, and NA. The preliminary results indicate that the bionic drug-containing CSF method has comparative advantages such as good reproducibility and high correlation between ingredients and effects in the in vitro neuropharmacological study of TCM.

Qingfei Paidu granules (QFPDG), derived from four classic traditional Chinese medicine formulas with a centuries-long history of treating respiratory disorders, have shown remarkable clinical efficacy against pneumonia and lung injury, yet their pharmacodynamic components and mechanisms require further elucidation. In this work, UPLC-Q-TOF-MS/MS was used to identify the chemical constituents in QFPDG, drug-containing serum, and lung tissue. Network pharmacology was employed to predict key targets and pathways. The anti-pneumonia efficacy was evaluated via an LPS-induced acute pneumonia mouse model and by measuring cytokine levels in LPS-stimulated RAW264.7 cells. Finally, Molecular docking along with molecular dynamics simulation was conducted to explore the interactions between key compounds and targets. Results showed that 145 compounds were identified in QFPDG solution, 94 in serum, and 83 in lung tissue, with 97 components in serum and lung associated with 350 pneumonia-related targets and crucial pathways. QFPDG alleviated acute lung injury and inflammation in LPS-induced acute pneumonia mice, reducing monocyte, neutrophil, lymphocyte, and leukocyte counts in bronchoalveolar lavage fluid (BALF), as well as decreasing TNF-α and IL-1β levels. Molecular docking and molecular dynamics simulations showed that five key components in QFPDG had strong binding affinities for TNF-α, IL-1β, IL-6 and AKT1 with minimal fluctuations at equilibrium. Binding free energy calculations indicated that the ∆G_bind values ranging from -8.59 to -41.98 kcal/mol, primarily driven by hydrophobic and electrostatic interactions, involving key amino acid residues like Glu93, Tyr97, Tyr119, Gly121, and Tyr151. In summary, QFPDG presents a multi-component, multi-target approach to treating pneumonia, targeting key signaling pathways including PI3K-AKT, MAPK, and TNF. The compound-target interactions clarify its anti-inflammatory and tissue-protective mechanisms, suggesting potential for clinical application and pharmaceutical development.