Development and Validation of a Novel Reverse Phase HPLC for Determination of Aloperine in Aloperine Tablets

Aim To develop a reverse phase HPLC method for the determination of aloperine, an alkaloid that is newly extracted from Sophora alopecuraides and has shown wide pharmacological effects including antibacterial and antiinflammatory actions. Methods The samples were analyzed on a ODS column with methanol water triethylamine (3∶97∶0 1 V/V) as a mobile phase. The flow rate was 1 0 mL·min -1 , and UV detection wavelength 205 nm. Results Linear regression equation was A=1 6920C+1 7455 (r 2=0 9999, n =5) in concentratins ranging from 20 to 120 μg·mL -1 . The recoveries were 101 2±1 46 % at 80 μg·mL -1 , 100 5±0 75% at 100 μg·mL -1 , and 100 7±1 10% at 120 μg·mL -1 , respectively, and the precisions of aloperine within or between run were from 0 80% to 1 98% ( n =5). The relative contents of aloperine in three lots of tablets were 101 59±1 38%, 98 46±0 23%, and 99 41±1 09% ( n =3). Conclusion The newly developed reverse phase HPLC method is simple and useful for daily assay of aloperine tablets and can overcome the interference from excipient and other alkaloids in titration and UV detection.

Chemical construction and anti-HCoV-OC43 evaluation of novel 10,12-disubstituted aloperine derivatives as dual cofactor inhibitors of TMPRSS2 and SR-B1

Thirty-one new 10,12-disubstituted aloperine derivatives were subtly constructed through a selective oxidation on the 10-α-C-H induced by sulfonyl and a nucleophilic substitution with the stereoselectivity and scalability.Of them,compound 6b displayed a moderate anti-human coronavirus OC43(HCoV-OC43)potency and blocked the viral entry stage through a host mechanism of action.Using chemoproteomic techniques,both transmembrane serine protease 2(TMPRSS2)and scavenger receptor class B type 1(SR-B1)proteins,which act as host cofactors of viral entry,were identified to be the direct targets of 6b against HCoV-OC43.Furthermore,6b may deactivate the TMPRSS2 by inducing a change in protein conformation,rather than binding to its catalytic center,thus suppressing the viral membrane fusion.Accordingly,our study provided key scientific data for the development of aloperine derivatives into a new class of antiviral candidates against humanβ-coronavirus,including severe acute respiratory syndrome coronavirus 2(SARS-CoV-2).

The anti-neoplastic activities of aloperine in HeLa cervical cancer cells are associated with inhibition of the IL-6-JAK1-STAT3 feedback loop

Cervical cancer(CC)is recognized as the most common neoplasm in the female reproductive system worldwide.The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients.Aloperine(ALO)is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation,allergies and infection.However,its therapeutic efficacy and underlying mechanism in CC are still unclear.In the current study,MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC.Then,the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry,respectively,while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay.Moreover,nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo.The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis.This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest.Simultaneously,ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio.Additionally,the migration and invasion of HeLa cells were attenuated by ALO treatment,which was considered to result from inhibition of epithelial-to-mesenchymal transition.For molecular mechanisms,the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment.This study indicated that ALO markedly suppresses the proliferation,migration and invasion and enhances the apoptosis of HeLa cells.In addition,these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.