Establishment of Withania somnifera Hairy Root Cultures for the Production of Withanolide A

Withania sominifera （Indian ginseng） was transformed by Agrobacterium rhizogenes. Explants from seedling roots, stems, hypocotyls, cotyledonary nodal segments, cotyledons and young leaves were inoculated with A. rhizogenes strain R1601. Hairy （transformed） roots were induced from cotyledons and leaf explants. The transgenic status of hairy roots was confirmed by polymerase chain reaction using nptll and rolB specific primers and, subsequently, by Southern analysis for the presence of nptll and rolB genes in the genomes of transformed roots. Four clones of hairy roots were established; these differed in their morphology. The doubling time of faster growing cultures was 8-14 d with a fivefold increase in biomass after 28 d compared with cultured, non-transformed seedling roots. MS-based liquid medium was superior for the growth of transformed roots compared with other culture media evaluated （SH, LS and N6）, with MS-based medium supplemented with 40 g/L sucrose being optimal for biomass production. Cultured hairy roots synthesized withanolide A, a steroidal lactone of medicinal and therapeutic value. The concentration of withanolide A in transformed roots （157.4 μg/g dry weight） was 2.7-fold more than in non-transformed cultured roots （57.9 μg/g dry weight）.

Withaminimas A–F, six withanolides with potential anti-inflammatory activity from Physalis minima

Withaminimas A–F(1–6), six new withaphysalin-type withanolides were isolated from the aerial parts of Physalis minima L.. The structures of these compounds were elucidated through a variety of spectroscopic techniques including HR-MS, NMR, and ECD. Compound 1 belongs to rare 18-norwithanolides, and 2–3 were 13/14-secowithanolides. According to the traditional usage of P. minima, inhibitory effects on nitric oxide(NO) production in lipopolysaccaride-activated RAW264.7 macrophages were evaluated, and compounds 1–4 exhibited significant inhibitory effects with IC50 values among 3.91–18.46 μmol·L–1.

NEW ANTIPROLIFERATIVE WITHANOLIDES FROM THE SEEDS OF DATURA METEL

Three new with anolides,daturaselines A-C(1-3),and three new with anolides glycosides,daturasesides A-C(4-6),were isolated and identified from ethyl acetate-soluble fraction of ethanol extract of Datura metel seeds.The structures of new compounds were established according to the 1D and 2D NMR spectra,combined with high resolution

A review of the ethnobotanical value,phytochemistry,and pharmacology of Physalis pubescens L.

Physalis pubescens L.(P.pubescens)was widely used for the treatment of inflammation-related diseases,such as sore throat,aphonia,phlegm,heat,and cough in folk medicine.The fruits of P.pubescens are commonly consumed as fruit in many areas of the world.In the past few decades,the phytochemistry and pharmacology of P.pubescens were extensively investigated.About 170 chemical constituents were purified from P.pubescens.The extract and chemical constituents of P.pubescens demonstrate diverse pharmacological effects,including anti-oxidation,anti-inflammation,anticancer,antimicrobial activity,immunomodulation,diuretic effect,hypoglycemic,and hypolipidemic in vitro and in vivo.Herein,we systematically summarized the ethnomedicinal uses,botanical characterization,distribution,phytochemistry,and pharmacology of P.pubescens,and establish the correlation between chemical constituents and pharmacological effects.

Identification of a novel PHGDH covalent inhibitor by chemical proteomics and phenotypic profiling

The first rate-limiting enzyme of the serine synthesis pathway(SSP), phosphoglycerate dehydrogenase(PHGDH), is hyperactive in multiple tumors, which leads to the activation of SSP and promotes tumorigenesis. However, only a few inhibitors of PHGDH have been discovered to date, especially the covalent inhibitors of PHGDH. Here, we identified withangulatin A(WA), a natural small molecule,as a novel covalent inhibitor of PHGDH. Affinity-based protein profiling identified that WA could directly bind to PHGDH and inactivate the enzyme activity of PHGDH. Biolayer interferometry and LC-MS/MS analysis further demonstrated the selective covalent binding of WA to the cysteine 295 residue(Cys295)of PHGDH. With the covalent modification of Cys295, WA blocked the substrate-binding domain(SBD)of PHGDH and exerted an allosteric effect to induce PHGDH inactivation. Further studies revealed that with the inhibition of PHGDH mediated by WA, the glutathione synthesis was decreased and intracellular levels of reactive oxygen species(ROS) were elevated, leading to the inhibition of tumor proliferation.This study indicates WA as a novel PHGDH covalent inhibitor, which identifies Cys295 as a novel allosteric regulatory site of PHGDH and holds great potential in developing anti-tumor agents for targeting PHGDH.