Isolation, Crystal Structure and Antitussive Activity of 9S,9aS-neotuberostemonine

The title compound 9S,9aS-neotuberostemonine （1） was isolated from the 95%ethanol extract of the roots of Stemona tuberosa. The crystal structure of 1, C22H33NO4, wasdetermined by single-crystal X-ray diffraction analysis. The crystal belongs to orthorhombicsystem, space group P212121, with a-9.0115（11）, b = 10.612（4）, c = 22.074（3） A, V= 2110.9（8）S^3, Z= 4, Mr= 375.49, Dc= 1.182 g/cm3, λ= 0.71079 A,μ= 0.080 cm^-1, F（000） = 816, S = 1.019,R = 0.0579 and wR = 0.1358. A total of 3109 unique reflections were collected, of which 2902were observed （I〉 2σ（I））. The absolute configuration of 1 could be assigned by referring to theconserved configuration of the methyl groups at C（13） and C（20）. In the solid state, the moleculeswere linked into a chain along the a-axis through weak hydrogen bond C（ll）-H（11A）…O（2）.Compound 1 shows significant inhibition of cough by 24%, 44% and 65% at doses of 50, 100 and150 mg/kg, respectively.

Anti-atherosclerotic effects and molecular targets of ginkgolide B from Ginkgo biloba

Bioactive compounds derived from herbal medicinal plants modulate various therapeutic targets and signaling pathways associated with cardiovascular diseases(CVDs),the world’s primary cause of death.Ginkgo biloba,a well-known traditional Chinese medicine with notable cardiovascular actions,has been used as a cardio-and cerebrovascular therapeutic drug and nutraceutical in Asian countries for centuries.Preclinical studies have shown that ginkgolide B,a bioactive component in Ginkgo biloba,can ameliorate atherosclerosis in cultured vascular cells and disease models.Of clinical relevance,several clinical trials are ongoing or being completed to examine the efficacy and safety of ginkgolide B-related drug preparations in the prevention of cerebrovascular diseases,such as ischemia stroke.Here,we present a comprehensive review of the pharmacological activities,pharmacokinetic characteristics,and mechanisms of action of ginkgolide B in atherosclerosis prevention and therapy.We highlight new molecular targets of ginkgolide B,including nicotinamide adenine dinucleotide phosphate oxidases(NADPH oxidase),lectin-like oxidized LDL receptor-1(LOX-1),sirtuin 1(SIRT1),platelet-activating factor(PAF),proprotein convertase subtilisin/kexin type 9(PCSK9)and others.Finally,we provide an overview and discussion of the therapeutic potential of ginkgolide B and highlight the future perspective of developing ginkgolide B as an effective therapeutic agent for treating atherosclerosis.

Deciphering chemical and metabolite profiling of Chang-Kang-Fang by UPLC-Q-TOF-MS/MS and its potential active components identification

Chang-Kang-Fang(CKF)formula,a Traditional Chinese Medicine(TCM)prescription,has been widely used for the treatment of irritable bowel syndrome(IBS).However,its potential material basis and underlying mechanism remain elusive.Therefore,this study employed an integrated approach that combined ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q/TOF-MS)with network pharmacology to systematically characterize the phytochemical components and metabolites of CKF,as well as elucidating its underlying mechanism.Through this comprehensive analysis,a total of 150 components were identified or tentatively characterized within the CKF formula.Notably,six N-acetyldopamine oligomers from Cicadae Periostracum and eight resin glycosides from Cuscutae Semen were characterized in this formula for the first time.Meanwhile,149 xenobiotics(58 prototypes and 91 metabolites)were detected in plasma,urine,feces,brain,and intestinal contents,and the in vivo metabolic pathways of resin glycosides were elaborated for the first time.Furthermore,network pharmacology and molecular docking analyses revealed that alkaloids,flavonoids,chromones,monoterpenes,N-acetyldopamine dimers,p-hydroxycinnamic acid,and Cus-3/isomer might be responsible for the beneficial effects of CKF in treating IBS,and CASP8,MARK14,PIK3C,PIK3R1,TLR4,and TNF may be its potential targets.These discoveries offer a comprehensive understanding of the potential material basis and clarify the underlying mechanism of the CKF formula in treating IBS,facilitating the broader application of CKF in the field of medicine.

Author correction to“The FAPα-activated prodrug Z-GP-DAVLBH inhibits the growth and pulmonary metastasis of osteosarcoma cells by suppressing the AXL pathway”[Acta Pharm Sin B 12(2022)1288–1304]

We deeply regret that the representative image of invaded 143B cells treated with Z-GP-DAVLBH + AXL-KD in the lower panel of Fig. 6C was incorrect due to inadvertent mistake of copying and pasting in the process of assembling figures with Adobe Illustrator software. The image has now been corrected, and the quantification of cell invasion in Fig. 6C remains correct. The corrected version of Fig. 6C has been provided below, and the change did not affect the results and conclusions of this study. We have provided the original data of this figure to Editorial Office, and the Editorial Office or the corresponding authors can be contacted for original data access.

Discovery and development of natural heat shock protein 90 inhibitors in cancer treatment

Heat shock protein 90(Hsp90)is a highly conserved molecular chaperone that plays a vital role in the signal transduction of cancers.Hsp90 inhibitors are able to inhibit Hsp90 or the complex of Hsp90 and co-chaperones resulting in the degradation of Hsp90-dependent client proteins through the ubiqui tina tion-proteasome pathway,thereby leading to the growth inhibition of tumor cells.This review will briefy discuss the molecular structure and biological function of Hsp90,and focus on a summary of recent progress in the development and testing of natural Hsp90 inhibitors and their different means by which they interact with Hsp90.

Absolute configuration of podophyllotoxone and its inhibitory activity against human prostate cancer cells

Podophyllotoxone(1) was isolated from the roots of Dysosma versipellis. The structure was determined by spectroscopic analysis in combination with single-crystal X-ray analysis. The absolute configuration of compound 1 was assigned based on the Flack parameter. It showed significant inhibitory activities against human prostate cancer cells PC3 and DU145 with IC50 values being 14.7 and 20.6 μmol·L-1, respectively. It also arrested the cells at G2/M phase. Tubulin polymerization assay showed that it inhibited the tubulin polymerization in a dose-dependent manner, and molecular docking analysis revealed a different binding mode with tubulin as compared with those known tubulin inhibitors.

The disbalance of LRP1 and SIRPα by psychological stress dampens the clearance of tumor cells by macrophages

The relationship between chronic psychological stress and tumorigenesis has been well defined in epidemiological studies;however, the underlying mechanism remains underexplored. In this study, we discovered that impaired macrophage phagocytosis contributed to the psychological stressevoked tumor susceptibility, and the stress hormone glucocorticoid(GC) was identified as a principal detrimental factor. Mechanistically, GC disturbed the balance of the "eat me" signal receptor(low-density lipoprotein receptor-related protein-1, LRP1) and the "don’t eat me" signal receptor(signal regulatory protein alpha, SIRPa). Further analysis revealed that GC led to a direct, glucocorticoid receptor(GR)-dependent trans-repression of LRP1 expression, and the repressed LRP1, in turn, resulted in the elevatedgene level of SIRPa by down-regulating mi RNA-4695-3 p. These data collectively demonstrate that stress induces the imbalance of the LRP1/SIRPa axis and entails the disturbance of tumor cell clearance by macrophages. Our findings provide the mechanistic insight into psychological stress-evoked tumor susceptibility and indicate that the balance of LRP1/SIRPa axis may serve as a potential therapeutic strategy for tumor treatment.

Maintaining the balance of TDP-43,mitochondria,and autophagy:a promising therapeutic strategy for neurodegenerative diseases

Mitochondria are the energy center of cell operations and are involved in physiological functions and maintenance of metabolic balance and homeostasis in the body.Alterations of mitochondrial function are associated with a variety of degenerative and acute diseases.As mitochondria age in cells,they gradually become inefficient and potentially toxic Acute injury can trigger the permeability of mitochondrial membranes,which can lead to apoptosis or necrosis.Transactive response DNA-binding protein 43 kDa(TDP-43)is a protein widely present in cells.It can bind to RNA,regulate a variety of RNA processes,and play a role in the formation of multi-protein/RNA complexes.Thus,the normal physiological functions of TDP-43 are particularly important for cell survival.Normal TDP-43 is located in various subcellular structures including mitochondria,mitochondrial-associated membrane,RNA particles and stress granules to regulate the endoplasmic reticulum-mitochondrial binding,mitochondrial protein translation,and mRNA transport and translation.Importantly,TDP-43 is associated with a variety of neurodegenerative diseases,including amyotrophic lateral sclerosis,frontotemporal dementia and Alzheimer's disease,which are characterized by abnormal phosphorylation,ubiquitination,lysis or nuclear depletion of TDP-43 in neurons and glial cells.Although the pathogenesis of TDP-43 proteinopathy remains unknown,the presence of pathological TDP-43 inside or outside of mitochondria and the functional involvement of TDP-43 in the regulation of mitochondrial morphology,transport,and function suggest that mitochondria are associated with TDP-43-related diseases.Autophagy is a basic physiological process that maintains the homeostasis of cells,including targeted clearance of abnormally aggregated proteins and damaged organelles in the cytoplasm;therefore,it is considered protective against neurodegenerative diseases.However,the combination of abnormal TDP-43 aggregation,mitochondrial dysfunction,and insufficient autophagy can lead to a variety of aging-related pathologies.In this review,we describe the current knowledge on the associations of mitochondria with TDP-43 and the role of autophagy in the clearance of abnormally aggregated TDP-43 and dysfunctional mitochondria.Finally,we discuss a novel approach for neurodegenerative treatment based on the knowledge.

The FAPα-activated prodrug Z-GP-DAVLBH inhibits the growth and pulmonary metastasis of osteosarcoma cells by suppressing the AXL pathway

Osteosarcoma is a kind of bone tumor with highly proliferative and invasive properties,a high incidence of pulmonary metastasis and a poor prognosis.Chemotherapy is the mainstay of treatment for osteosarcoma.Currently,there are no molecular targeted drugs approved for osteosarcoma treatment,particularly effective drugs for osteosarcoma with pulmonary metastases.It has been reported that fibroblast activation protein alpha(FAPa)is upregulated in osteosarcoma and critically associated with osteosarcoma progression and metastasis,demonstrating that FAPa-targeted agents might be a promising therapeutic strategy for osteosarcoma.In the present study,we reported that the FAPa-activated vinblastine prodrug Z-GP-DAVLBH exhibited potent antitumor activities against FAPa-positive osteosarcoma cells in vitro and in vivo.Z-GP-DAVLBH inhibited the growth and induced the apoptosis of osteosarcoma cells.Importantly,it also decreased the migration and invasion capacities and reversed epithelial-mesenchymal transition(EMT)of osteosarcoma cells in vitro and suppressed pulmonary metastasis of osteosarcoma xenografts in vivo.Mechanistically,Z-GP-DAVLBH suppressed the AXL/AKT/GSK-3β/β-catenin pathway,leading to inhibition of the growth and metastatic spread of osteosarcoma cells.These findings demonstrate that Z-GP-DAVLBH is a promising agent for the treatment of FAPa-positive osteosarcoma,particularly osteosarcoma with pulmonary metastases.

Discovery of a novel EGFR ligand DPBA that degrades EGFR and suppresses EGFR-positive NSCLC growth

Epidermal growth factor receptor(EGFR)activation plays a pivotal role in EGFR-driven non-small cell lung cancer(NSCLC)and is considered as a key target of molecular targeted therapy.EGFR tyrosine kinase inhibitors(TKIs)have been canonically used in NSCLC treatment.However,prevalent innate and acquired resistances and EGFR kinase-independent pro-survival properties limit the clinical efficacy of EGFR TKIs.Therefore,the discovery of novel EGFR degraders is a promising approach towards improving therapeutic efficacy and overcoming drug resistance.Here,we identified a 23-hydroxybetulinic acid derivative,namely DPBA,as a novel EGFR small-molecule ligand.It exerted potent in vitro and in vivo anticancer activity in both EGFR wild type and mutant NSCLC by degrading EGFR.Mechanistic studies disclosed that DPBA binds to the EGFR extracellular domain at sites differing from those of EGF and EGFR.DPBA did not induce EGFR dimerization,phosphorylation,and ubiquitination,but it significantly promoted EGFR degradation and repressed downstream survival pathways.Further analyses showed that DPBA induced clathrin-independent EGFR endocytosis mediated by flotillin-dependent lipid rafts and unaffected by EGFR TKIs.Activation of the early and late endosome markers rab5 and rab7 but not the recycling endosome marker rab11 was involved in DPBA-induced EGFR lysosomal degradation.The present study offers a new EGFR ligand for EGFR pharmacological degradation and proposes it as a potential treatment for EGFR-positive NSCLC,particularly NSCLC with innate or acquired EGFR TKI resistance.DPBA can also serve as a chemical probe in the studies on EGFR trafficking and degradation.

Synthesis of Camptothecin-lO-O-glucoside Using an Engineered Oleandomycin Glucosyltransferase

Oleandomycin glycosyltransferase variant P67T/S132F/A242V（ASP） was used to convert 10-hydroxycamptothecin into camptothecin-10-O-glucoside, which was confirmed by spectral analysis. Compared to the previously reported results, the present study reached the conversion rate up to 80% through the optimization of reaction conditions. In addition, compared with 10-hydroxycamptotheein（HCPT）, camptothecin-10-O-glucoside inhibited the proliferation of Huh7 cells in a concentration-dependent manner and showed stronger antineoplastic effect but lower toxicity. Furthermore, camptothecin-10-O-glucoside induced more apoptotic cells as compared with the parent compound.

Isolation, Crystal Structure and Cytotoxic Activity of Natural Maistemonine and Comparison with the Synthetic Compound

The title compound maistemonine(1)was isolated from the total alkaloid fraction of the 95%ethanol extract of the roots of Stemona japonica,followed by preparative HPLC and recrystallization from a mixture of n-hexane and ethyl acetate.The crystal structure of 1,C23H29NO)6,was determined by single-crystal X-ray diffraction analysis.The crystal belongs to orthorhombic system,space group P212121 with a=8.5698(6),b=14.0460(11),c=17.8815(17)?,V=2152.4(3)?3,Z=4,Mr=415.47,Dc=1.282 g/cm3,λ=0.71079?,μ=0.092 cm^–1,F(000)=888,S=0.995,R=0.0535 and wR=0.1067.A total of 5136 unique reflections were collected,of which 3523 were observed(Ⅰ>2σ(Ⅰ)).The absolute configuration of 1 could be assigned by referring to the conserved configuration of the methyl groups at C(2).Compound 1 shows mild cytotoxic activity against the prostate cancer cells LNCaP and PC3 with the IC50 values of 29.4±2.3 and 46.6±3.1 mM,respectively.It is noteworthy that the natural maistemonine(1)is different from the synthetic compound which was a racemic with the triclinic space group P1.The lactone rings E from the natural and synthetic 1 are almost perpendicular to each other when overlapping the remaining parts of the molecules.

Targeting FAPα-positive lymph node metastatic tumor cells suppresses colorectal cancer metastasis

Lymphatic metastasis is the main metastatic route for colorectal cancer, which increases the risk of cancer recurrence and distant metastasis. The properties of the lymph node metastatic colorectal cancer(LNM-CRC) cells are poorly understood, and effective therapies are still lacking. Here, we found that hypoxia-induced fibroblast activation protein alpha(FAPα) expression in LNM-CRC cells. Gain-or loss-function experiments demonstrated that FAPα enhanced tumor cell migration, invasion, epithelial-mesenchymal transition, stemness, and lymphangiogenesis via activation of the STAT3 pathway. In addition, FAPα in tumor cells induced extracellular matrix remodeling and established an immunosuppressive environment via recruiting regulatory T cells, to promote colorectal cancer lymph node metastasis(CRCLNM). Z-GP-DAVLBH, a FAPα-activated prodrug, inhibited CRCLNM by targeting FAPα-positive LNM-CRC cells. Our study highlights the role of FAPα in tumor cells in CRCLNM and provides a potential therapeutic target and promising strategy for CRCLNM.

Chemical screen identifies shikonin as a broad DNA damage response inhibitor that enhances chemotherapy through inhibiting ATM and ATR

DNA damage response(DDR)is a highly conserved genome surveillance mechanism that preserves cell viability in the presence of chemotherapeutic drugs.Hence,small molecules that inhibit DDR are expected to enhance the anti-cancer effect of chemotherapy.Through a recent chemical library screen,we identified shikonin as an inhibitor that strongly suppressed DDR activated by various chemotherapeutic drugs in cancer cell lines derived from different origins.Mechanistically,shikonin inhibited the activation of ataxia telangiectasia mutated(ATM),and to a lesser degree ATM and RAD3-related(ATR),two master upstream regulators of the DDR signal,through inducing degradation of ATM and ATR-interacting protein(ATRIP),an obligate associating protein of ATR,respectively.As a result of DDR inhibition,shikonin enhanced the anti-cancer effect of chemotherapeutic drugs in both cell cultures and in mouse models.While degradation of ATRIP is proteasome dependent,that of ATM depends on caspase-and lysosome-,but not proteasome.Overexpression of ATM significantly mitigated DDR inhibition and cell death induced by shikonin and chemotherapeutic drugs.These novel findings reveal shikonin as a pan DDR inhibitor and identify ATM as a primary factor in determining the chemo sensitizing effect of shikonin.Our data may facilitate the development of shikonin and its derivatives as potential chemotherapy sensitizers through inducing ATM degradation.