ABSTRACT
We formerly reported that EZH2 inhibitors sensitized HIF-1 inhibitor-resistant cells and inhibited HIF-1α to promote SUZ12 transcription, leading to enhanced EZH2 enzyme activity and elevated H3K27me3 levels, and conversely, inhibition of EZH2 promoted HIF-1α transcription. HIF-1α and EZH2 interacted to form a negative feedback loop that reinforced each other's activity. In this paper, a series of 2,2- dimethylbenzopyran derivatives containing pyridone structural fragments were designed and synthesized with DYB-03, a HIF-1α inhibitor previously reported by our group, and Tazemetostat, an EZH2 inhibitor approved by FDA, as lead compounds. Among these compounds, D-01 had significant inhibitory activities on HIF-1α and EZH2. In vitro experiments showed that D-01 significantly inhibited the migration of A549 cells, clone, invasion and angiogenesis. Moreover, D-01 had good pharmacokinetic profiles. All the results about compound D-01 could lay a foundation for the research and development of HIF-1α and EZH2 dual-targeting compounds.
Subject(s)
Antineoplastic Agents , Drug Screening Assays, Antitumor , Enhancer of Zeste Homolog 2 Protein , Hypoxia-Inducible Factor 1, alpha Subunit , Lung Neoplasms , Pyridones , Humans , Enhancer of Zeste Homolog 2 Protein/antagonists & inhibitors , Enhancer of Zeste Homolog 2 Protein/metabolism , Hypoxia-Inducible Factor 1, alpha Subunit/antagonists & inhibitors , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Pyridones/chemistry , Pyridones/pharmacology , Pyridones/chemical synthesis , Lung Neoplasms/drug therapy , Lung Neoplasms/pathology , Lung Neoplasms/metabolism , Structure-Activity Relationship , Antineoplastic Agents/pharmacology , Antineoplastic Agents/chemistry , Antineoplastic Agents/chemical synthesis , Molecular Structure , Dose-Response Relationship, Drug , Cell Proliferation/drug effects , Animals , Benzopyrans/chemistry , Benzopyrans/pharmacology , Benzopyrans/chemical synthesis , Cell Movement/drug effectsABSTRACT
DNA G-quadruplex(G4) is a guanine-rich single-stranded DNA sequence that spontaneously folds into a spherical four-stranded DNA secondary structure in oncogene promoter sequences and telomeres. G4s are highly associated with the occurrence and development of cancer and have emerged as promising anticancer targets. Natural products have long been important sources of anticancer drug development. In recent years, significant progress has been made in the discovery of natural drugs targeting DNA G4s, with many DNA G4s have been confirmed as promising targets of natural products, including MYC-G4, KRAS-G4, PDGFR-ß-G4, BCL-2-G4, VEGF-G4, and telomeric G4. This review summarizes the research progress in discovering natural small molecules that target DNA G4s and their binding mechanisms. It also discusses the opportunities of and challenges in developing drugs targeting DNA G4s. This review will serve as a valuable reference for the research on natural products, particularly in the development of novel antitumor medications.
Subject(s)
Biological Products , G-Quadruplexes , G-Quadruplexes/drug effects , Biological Products/chemistry , Biological Products/pharmacology , Humans , Animals , DNA/chemistry , DNA/genetics , Neoplasms/drug therapy , Neoplasms/genetics , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacologyABSTRACT
EGFR tyrosine kinase inhibitors have made remarkable success in targeted cancer therapy. However, therapeutic resistance inevitably occurred and EGFR-targeting therapy has been demonstrated to have limited efficacy or utility in glioblastoma, colorectal cancer, and hepatocellular carcinoma. Therefore, there is a high demand for the development of new targets to inhibit EGFR signaling. Herein, we found that the EGFR oncogene proximal promoter sequence forms a unique type of snap-back loop containing G-quadruplex (G4), which can be targeted by small molecules. For the first time, we determined the NMR solution structure of this snap-back EGFR-G4, a three-tetrad-core, parallel-stranded G4 with naturally occurring flanking residues at both the 5'-end and 3'-end. The snap-back loop located at the 3'-end region forms a stable capping structure through two stacked G-triads connected by multiple potential hydrogen bonds. Notably, the flanking residues are consistently absent in reported snap-back G4s, raising the question of whether such structures truly exist under in vivo conditions. The resolved EGFR-G4 structure has eliminated the doubt and showed distinct structural features that distinguish it from the previously reported snap-back G4s, which lack the flanking residues. Furthermore, we found that the snap-back EGFR-G4 structure is highly stable and can form on an elongated DNA template to inhibit DNA polymerase. The unprecedented high-resolution EGFR-G4 structure has thus contributed a promising molecular target for developing alternative EGFR signaling inhibitors in cancer therapeutics. Meanwhile, the two stacked triads may provide an attractive site for specific small-molecule targeting.
Subject(s)
G-Quadruplexes , Neoplasms , Humans , Promoter Regions, Genetic , Oncogenes , ErbB Receptors/geneticsABSTRACT
Guanine (G)-oxidation to 8-oxo-7,8-dihydroguanine (OG) by reactive oxygen species in genomic DNA has been implicated with various human diseases. G-quadruplex (G4)-forming sequences in gene promoters are highly susceptible to G-oxidation, which can subsequently cause gene activation. However, the underlying G4 structural changes that result from OG modifications remain poorly understood. Herein, we investigate the effect of G-oxidation on the BLM gene promoter G4. For the first time, we show that OG can induce a G-vacancy-containing G4 (vG4), which can be filled in and stabilized by guanine metabolites and derivatives. We determined the NMR solution structure of the cGMP-fill-in oxidized BLM promoter vG4. This is the first complex structure of an OG-induced vG4 from a human gene promoter sequence with a filled-in guanine metabolite. The high-resolution structure elucidates the structural features of the specific 5'-end cGMP-fill-in for the OG-induced vG4. Interestingly, the OG is removed from the G-core and becomes part of the 3'-end capping structure. A series of guanine metabolites and derivatives are evaluated for fill-in activity to the oxidation-induced vG4. Significantly, cellular guanine metabolites, such as cGMP and GTP, can bind and stabilize the OG-induced vG4, suggesting their potential regulatory role in response to oxidative damage in physiological and pathological processes. Our work thus provides exciting insights into how oxidative damage and cellular metabolites may work together through a G4-based epigenetic feature for gene regulation. Furthermore, the NMR structure can guide the rational design of small-molecule inhibitors that specifically target the oxidation-induced vG4s.
Subject(s)
G-Quadruplexes , Guanine , Guanine/chemistry , Humans , Oxidation-Reduction , Oxidative Stress , Promoter Regions, GeneticABSTRACT
Two pairs of unprecedented ß-carboline-phenylpropanoid heterogeneous alkaloids, (±)-pheharmines A-B (1-4), characterized by a morpholino[4,3,2-hi]ß-carboline core with two chiral centers, were isolated from the roots of Peganum harmala. The structures, including their absolute configurations, were identified using spectroscopic analyses and electronic circular dichroism (ECD) calculations. The biosynthetic hypothesis for the formation of pheharmines A-B was proposed. Compounds 1-4 exhibited moderate cytotoxic activities against HL-60 cell lines.
Subject(s)
Alkaloids , Peganum , Humans , Peganum/chemistry , Peganum/metabolism , Morpholinos/analysis , Morpholinos/metabolism , Seeds , Molecular Structure , Alkaloids/pharmacology , Alkaloids/chemistry , Carbolines/pharmacology , Carbolines/chemistryABSTRACT
Soil degradation is significantly increased driven by soil nutrient loss and soil erodibility, thus, hampering the sustainable development of the ecological environment and agricultural production. Vegetation restoration has been widely adopted to prevent soil degradation given its role in improving soil nutrients and soil erodibility. However, it is unclear which vegetation type has the best improving capacity from soil nutrient and soil erodibility perspectives. This study selected three vegetation restoration types of grasslands (GL), shrublands (SL), and forestlands (FL) along the five slope positions (i.e., top, upper, middle, lower, and foot slope), to investigate the effects of vegetation restoration types on soil nutrients and soil erodibility. All vegetation restoration types were restored for 20 years from croplands (CL). We used comprehensive soil nutrient index (CSNI) and comprehensive soil erodibility index (CSEI) formed by a weighted summation method to reflect the effect of vegetation restoration on the improving capacity of soil nutrient and erodibility. The results showed the vegetation types with the highest comprehensive soil quality index (CSQI) at the top, upper, middle, lower and foot slope were FL (1.92), FL (1.98), SL (2.15), FL (2.37) and GL (3.93), respectively. When only one vegetation type was considered on the entire slope, SL (0.59) and FL (0.59) had the highest CSNI, the SL had the lowest CSEI (0.34) and the highest CSQI (1.89). The CSNI was mainly influenced by soil structure stability index (SSSI), sand content, silt + clay particles, and CSEI was controlled by soil organic matter (SOM), macroaggregates and microaggregates. Moreover, the CSQI was influenced by pH, silt and clay content, and biome coverage (BC). The study suggested the SL were advised as the best vegetation restoration type on the whole slope from improving soil nutrients and soil erodibility.
Subject(s)
Forests , Soil , Agriculture , China , Ecosystem , NutrientsABSTRACT
The G-quadruplexes (G4s) formed in the PDGFR-ß gene promoter are transcriptional modulators and amenable to small-molecule targeting. Berberine (BER), a clinically important natural isoquinoline alkaloid, has gained increasing attention due to its potential as anticancer drug. We previously showed that the PDGFR-ß gene promoter forms a unique vacancy G4 (vG4) that can be filled in and stabilized by guanine metabolites, such as dGMP. Herein, we report the high-resolution NMR structure of a ternary complex of berberine bound to the dGMP-fill-in PDGFR-ß vG4 in potassium solution. This is the first small-molecule complex structure of a fill-in vG4. This ternary complex has a 2:1:1 binding stoichiometry with a berberine molecule bound at each the 5'- and 3'-end of the 5'-dGMP-fill-in PDGFR-ß vG4. Each berberine recruits the adjacent adenine residue from the 5'- or 3'-flanking sequence to form a "quasi-triad plane" that covers the external G-tetrad of the fill-in vG4, respectively. Significantly, berberine covers and stabilizes the fill-in dGMP. The binding of berberine involves both π-stacking and electrostatic interactions, and the fill-in dGMP is covered and well-protected by berberine. The NMR structure can guide rational design of berberine analogues that target the PDGFR-ß vG4 or dGMP-fill-in vG4. Moreover, our structure provides a molecular basis for designing small-molecule guanine conjugates to target vG4s.
Subject(s)
G-QuadruplexesABSTRACT
G-quadruplexes are four-stranded nucleic acid secondary structures of biological significance and have emerged as an attractive drug target. The G4 formed in the MYC promoter (MycG4) is one of the most studied small-molecule targets, and a model system for parallel structures that are prevalent in promoter DNA G4s and RNA G4s. Molecular docking has become an essential tool in structure-based drug discovery for protein targets, and is also increasingly applied to G4 DNA. However, DNA, and in particular G4, binding sites differ significantly from protein targets. Here we perform the first systematic evaluation of four commonly used docking programs (AutoDock Vina, DOCK 6, Glide, and RxDock) for G4 DNA-ligand binding pose prediction using four small molecules whose complex structures with the MycG4 have been experimentally determined in solution. The results indicate that there are considerable differences in the performance of the docking programs and that DOCK 6 with GB/SA rescoring performs better than the other programs. We found that docking accuracy is mainly limited by the scoring functions. The study shows that current docking programs should be used with caution to predict G4 DNA-small molecule binding modes.
Subject(s)
DNA/metabolism , G-Quadruplexes , Molecular Docking Simulation , Proto-Oncogene Proteins c-myc/metabolism , Small Molecule Libraries/metabolism , Software , Binding Sites , DNA/chemistry , DNA/genetics , Humans , Ligands , Proto-Oncogene Proteins c-myc/geneticsABSTRACT
Aberrant expression of PDGFR-ß is associated with a number of diseases. The G-quadruplexes (G4s) formed in PDGFR-ß gene promoter are transcriptional modulators and amenable to small molecule targeting. The major G4 formed in the PDGFR-ß gene promoter was previously shown to have a broken G-strand. Herein, we report that the PDGFR-ß gene promoter sequence forms a vacancy G-quadruplex (vG4) which can be filled in and stabilized by physiologically relevant guanine metabolites, such as dGMP, GMP, and cGMP, as well as guanine-derivative drugs. We determined the NMR structure of the dGMP-fill-in PDGFR-ß vG4 in K+ solution. This is the first structure of a guanine-metabolite-fill-in vG4 based on a human gene promoter sequence. Our structure and systematic analysis elucidate the contributions of Hoogsten hydrogen bonds, sugar, and phosphate moieties to the specific G-vacancy fill-in. Intriguingly, an equilibrium of 3'- and 5'-end vG4s is present in the PDGFR-ß promoter sequence, and dGMP favors the 5'-end fill-in. Guanine metabolites and drugs were tested and showed a conserved selectivity for the 5'-vacancy, except for cGMP. cGMP binds both the 3'- and 5'-end vG4s and forms two fill-in G4s with similar population. Significantly, guanine metabolites are involved in many physiological and pathological processes in human cells; thus, our results provide a structural basis to understand their potential regulatory functions by interaction with promoter vG4s. Moreover, the NMR structure can guide rational design of ligands that target the PDGFR-ß vG4.
Subject(s)
DNA/metabolism , Deoxyguanine Nucleotides/metabolism , G-Quadruplexes , Promoter Regions, Genetic , Receptor, Platelet-Derived Growth Factor beta/genetics , DNA/genetics , Humans , Nuclear Magnetic Resonance, BiomolecularABSTRACT
The effects of nitrogen (N) deposition on soil organic carbon (C) and greenhouse gas (GHG) emissions in terrestrial ecosystems are the main drivers affecting GHG budgets under global climate change. Although many studies have been conducted on this topic, we still have little understanding of how N deposition affects soil C pools and GHG budgets at the global scale. We synthesized a comprehensive dataset of 275 sites from multiple terrestrial ecosystems around the world and quantified the responses of the global soil C pool and GHG fluxes induced by N enrichment. The results showed that the soil organic C concentration and the soil CO2 , CH4 and N2 O emissions increased by an average of 3.7%, 0.3%, 24.3% and 91.3% under N enrichment, respectively, and that the soil CH4 uptake decreased by 6.0%. Furthermore, the percentage increase in N2 O emissions (91.3%) was two times lower than that (215%) reported by Liu and Greaver (Ecology Letters, 2009, 12:1103-1117). There was also greater stimulation of soil C pools (15.70 kg C ha-1 year-1 per kg N ha-1 year-1 ) than previously reported under N deposition globally. The global N deposition results showed that croplands were the largest GHG sources (calculated as CO2 equivalents), followed by wetlands. However, forests and grasslands were two important GHG sinks. Globally, N deposition increased the terrestrial soil C sink by 6.34 Pg CO2 /year. It also increased net soil GHG emissions by 10.20 Pg CO2 -Geq (CO2 equivalents)/year. Therefore, N deposition not only increased the size of the soil C pool but also increased global GHG emissions, as calculated by the global warming potential approach.
ABSTRACT
Two novel cucurbitane 3-nor-triterpenoids, named norcolocynthenins A (1) and B (2), were isolated from the fruits of Citrullus colocynthis. The structures including their absolute configurations were determined by extensive spectroscopic analyses and theoretical calculations. Compound 1 features an unprecedented 5/6/6/5-fused ring system while compound 2 possesses a rare lactone moiety at modified ring A. Compounds 1 and 2 showed significant cytotoxic activity against human cancer cell lines of HL-60 (IC50 = 8.32, 6.49 µM) and PC-3 (IC50 = 31.26, 13.42 µM). The plausible biosynthetic pathway of compounds 1 and 2 via a key enzymatic Baeyer-Villiger reaction is proposed.
Subject(s)
Citrullus colocynthis/chemistry , Glycosides/chemistry , Triterpenes/chemistry , Animals , Humans , Male , Molecular Structure , Rats , Rats, Sprague-DawleyABSTRACT
MYC is one of the most important oncogenes and is overexpressed in the majority of cancers. G-Quadruplexes are noncanonical four-stranded DNA secondary structures that have emerged as attractive cancer-specific molecular targets for drug development. The G-quadruplex formed in the proximal promoter region of the MYC oncogene (MycG4) has been shown to be a transcriptional silencer that is amenable to small-molecule targeting for MYC suppression. Indenoisoquinolines are human topoisomerase I inhibitors in clinical testing with improved physicochemical and biological properties as compared to the clinically used camptothecin anticancer drugs topotecan and irinotecan. However, some indenoisoquinolines with potent anticancer activity do not exhibit strong topoisomerase I inhibition, suggesting a separate mechanism of action. Here, we report that anticancer indenoisoquinolines strongly bind and stabilize MycG4 and lower MYC expression levels in cancer cells, using various biochemical, biophysical, computer modeling, and cell-based methods. Significantly, a large number of active indenoisoquinolines cause strong MYC downregulation in cancer cells. Structure-activity relationships of MycG4 recognition by indenoisoquinolines are investigated. In addition, the analysis of indenoisoquinoline analogues for their MYC-inhibitory activity, topoisomerase I-inhibitory activity, and anticancer activity reveals a synergistic effect of MYC inhibition and topoisomerase I inhibition on anticancer activity. Therefore, this study uncovers a novel mechanism of action of indenoisoquinolines as a new family of drugs targeting the MYC promoter G-quadruplex for MYC suppression. Furthermore, the study suggests that dual targeting of MYC and topoisomerase I may serve as a novel strategy for anticancer drug development.
Subject(s)
DNA Topoisomerases, Type I/metabolism , Down-Regulation/drug effects , Indenes/pharmacology , Isoquinolines/pharmacology , Proto-Oncogene Proteins c-myc/antagonists & inhibitors , Topoisomerase I Inhibitors/pharmacology , Binding Sites/drug effects , G-Quadruplexes/drug effects , Humans , Indenes/chemistry , Isoquinolines/chemistry , Molecular Structure , Promoter Regions, Genetic/drug effects , Promoter Regions, Genetic/genetics , Protein Stability/drug effects , Proto-Oncogene Proteins c-myc/genetics , Proto-Oncogene Proteins c-myc/metabolism , Structure-Activity Relationship , Topoisomerase I Inhibitors/chemistryABSTRACT
Baicalein and scutellarein are bioactive flavones found in the medicinal plant Scutellaria baicalensis Georgi, used in traditional Chinese medicine. Extensive previous work has demonstrated the broad biological activity of these flavonoids, such as antifibrotic, antiviral and anticancer properties. However, their supply from plant material is insufficient to meet demand. Here, to provide an alternative production source and increase production levels of these flavones, we engineered an artificial pathway in an Escherichia coli cell factory for the first time. By first reconstructing the plant flavonoid biosynthetic pathway genes from five different species: phenylalanine ammonia lyase from Rhodotorula toruloides (PAL), 4-coumarate-coenzyme A ligase from Petroselinum crispum (4CL), chalcone synthase from Petunia hybrida (CHS), chalcone isomerase from Medicago sativa (CHI) and an oxidoreductase flavone synthase I from P. crispum (FNSI), production of the intermediates chrysin and apigenin was achieved by feeding phenylalanine and tyrosine as precursors. By comparative analysis of various versions of P450s, a construction expressing 2B1 incorporated with a 22-aa N-terminal truncated flavone C-6 hydroxylase from S. baicalensis (F6H) and partner P450 reductase from Arabidopsis thaliana (AtCPR) was found most effective for production of both baicalein (8.5â¯mg/L) and scutellarein (47.1â¯mg/L) upon supplementation with 0.5â¯g/L phenylalanine and tyrosine in 48â¯h of fermentation. Finally, optimization of malonyl-CoA availability further increased the production of baicalein to 23.6â¯mg/L and scutellarein to 106.5â¯mg/L in a flask culture. This report presents a significant advancement of flavone synthetic production and provides foundation for production of other flavones in microbial hosts.
Subject(s)
Apigenin/biosynthesis , Escherichia coli/genetics , Escherichia coli/metabolism , Flavanones/biosynthesis , Metabolic Engineering/methods , Phenylalanine/metabolism , Plants/metabolism , Tyrosine/metabolism , Biosynthetic Pathways/genetics , Flavonoids/biosynthesis , Malonyl Coenzyme A/metabolism , Scutellaria baicalensisABSTRACT
The human telomeric G-quadruplex (G4) is an attractive target for developing anticancer drugs. Natural products protoberberine alkaloids are known to bind human telomeric G4 and inhibit telomerase. Among several structurally similar protoberberine alkaloids, epiberberine (EPI) shows the greatest specificity in recognizing the human telomeric G4 over duplex DNA and other G4s. Recently, NMR study revealed that EPI recognizes specifically the hybrid-2 form human telomeric G4 by inducing large rearrangements in the 5'-flanking segment and loop regions to form a highly extensive four-layered binding pocket. Using the NMR structure of the EPI-human telomeric G4 complex, here we perform molecular dynamics free energy calculations to elucidate the ligand selectivity in the recognition of protoberberines by the human telomeric G4. The MM-PB(GB)SA (molecular mechanics-Poisson Boltzmann/Generalized Born) Surface Area) binding free energies calculated using the Amber force fields bsc0 and OL15 correlate well with the NMR titration and binding affinity measurements, with both calculations correctly identifying the EPI as the strongest binder to the hybrid-2 telomeric G4 wtTel26. The results demonstrated that accounting for the conformational flexibility of the DNA-ligand complexes is crucially important for explaining the ligand selectivity of the human telomeric G4. While the MD-simulated (molecular dynamics) structures of the G-quadruplex-alkaloid complexes help rationalize why the EPI-G4 interactions are optimal compared with the other protoberberines, structural deviations from the NMR structure near the binding site are observed in the MD simulations. We have also performed binding free energy calculation using the more rigorous double decoupling method (DDM); however, the results correlate less well with the experimental trend, likely due to the difficulty of adequately sampling the very large conformational reorganization in the G4 induced by the protoberberine binding.
Subject(s)
Berberine Alkaloids/chemistry , G-Quadruplexes/drug effects , Telomere/chemistry , Binding Sites , Fluorescence , Humans , Hydrogen Bonding , Ligands , Magnetic Resonance Spectroscopy , Molecular Dynamics Simulation , Molecular Structure , Structure-Activity Relationship , ThermodynamicsABSTRACT
Seventeen quinazoline alkaloids and derivatives, containing two pairs of new epimers, named as (S)- and (R)-1-(2-aminobenzyl)-3-hydroxypyrrolidin-2-one ß-d-glucopyranosyl-(1â¯ââ¯6)-ß-d-glucopyranoside (1, 2), (S)- and (R)-vasicinone ß-d-glucopyranosyl-(1â¯ââ¯6)-ß-d-glucopyranoside (3, 4), and a new enantiomer (12b), together with six known ones (5-8, 10, and 12a), and three pairs of known enantiomers (9, 11, and 13), were isolated from the ethanol extracts of the seeds of Peganum harmala L.. Their structures including the absolute configuration were elucidated by using 1D and 2D NMR, and ECD calculation approaches. The cytotoxic activities of all isolated compounds were evaluated. 11 showed moderate cytotoxicity against PC-3 cells with an IC50 value of 15.41⯵M.
Subject(s)
Alkaloids/pharmacology , Antineoplastic Agents, Phytogenic/pharmacology , Peganum/chemistry , Quinazolines/pharmacology , Seeds/chemistry , Alkaloids/chemistry , Alkaloids/isolation & purification , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Cell Line, Tumor , Cell Proliferation/drug effects , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Humans , Molecular Structure , Quinazolines/chemistry , Quinazolines/isolation & purification , Structure-Activity RelationshipABSTRACT
Human telomeres can form DNA G-quadruplex (G4), an attractive target for anticancer drugs. Human telomeric G4s bear inherent structure polymorphism, challenging for understanding specific recognition by ligands or proteins. Protoberberines are medicinal natural-products known to stabilize telomeric G4s and inhibit telomerase. Here we report epiberberine (EPI) specifically recognizes the hybrid-2 telomeric G4 predominant in physiologically relevant K+ solution and converts other telomeric G4 forms to hybrid-2, the first such example reported. Our NMR structure in K+ solution shows EPI binding induces extensive rearrangement of the previously disordered 5'-flanking and loop segments to form an unprecedented four-layer binding pocket specific to the hybrid-2 telomeric G4; EPI recruits the (-1) adenine to form a "quasi-triad" intercalated between the external tetrad and a T:T:A triad, capped by a T:T base pair. Our study provides structural basis for small-molecule drug design targeting the human telomeric G4.
Subject(s)
Berberine/analogs & derivatives , G-Quadruplexes/drug effects , Telomere , Base Sequence , Berberine/metabolism , Berberine/pharmacology , Binding Sites , Circular Dichroism , Drugs, Chinese Herbal , Humans , Intercalating Agents/chemistry , Nucleic Acid Conformation , Potassium/chemistry , Proton Magnetic Resonance SpectroscopyABSTRACT
Neobraclactones A-C (1-3), featuring an unprecedented further rearranged prenylxanthone skeleton with a unique octahydro-2H-1,3-dioxacyclopenta[c,d]inden-2-one scaffold, along with their biosynthesis-related known compound neobractatin (4), were isolated from the leaves of Garcinia bracteata. Their structures with absolute configurations were determined by extensive analyses of spectroscopic data and ECD calculations. Compounds 1 and 2 showed significant growth inhibition activities against the human leukaemia HL-60 and K562 cell lines with GI50 values from 0.40 to 0.86 µM.
Subject(s)
Garcinia/chemistry , Xanthones/isolation & purification , Molecular Conformation , Xanthones/chemistryABSTRACT
Inspired by the intriguing structures and bioactivities of dimeric alkaloids, 11 new thalifaberine-type aporphine-benzylisoquinoline alkaloids, thalicultratines A-K, a tetrahydroprotoberberine-aporphine alkaloid, thalicultratine L, and five known ones were isolated from the roots of Thalictrum cultratum. Their structures were defined on the basis of NMR and HRESIMS data. The antiproliferative activities of compounds 1-17 were evaluated against human leukemia HL-60 and prostate cancer PC-3 cells. Most alkaloids showed potent cytotoxicity against selected cancer cells. Preliminary SARs are discussed. The most active new compound (3), with an IC50 value of 1.06 µM against HL-60 cells, was selected for mechanism of action studies. The results revealed that compound 3 induced apoptosis and arrested the HL-60 cell cycle at the S phase with the loss of mitochondria membrane potential. The nuclear morphological Hoechst 33258 staining assay was also carried out, and the results confirmed apoptosis.
Subject(s)
Alkaloids/isolation & purification , Alkaloids/pharmacology , Antineoplastic Agents, Phytogenic/isolation & purification , Antineoplastic Agents, Phytogenic/pharmacology , Aporphines/isolation & purification , Aporphines/pharmacology , Drugs, Chinese Herbal/isolation & purification , Drugs, Chinese Herbal/pharmacology , Plant Roots/chemistry , Thalictrum/chemistry , Alkaloids/chemistry , Antineoplastic Agents, Phytogenic/chemistry , Aporphines/chemistry , Berberine Alkaloids , Drugs, Chinese Herbal/chemistry , HL-60 Cells , Humans , Inhibitory Concentration 50 , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Structure-Activity RelationshipABSTRACT
Investigation of the alkaloids from Peganum harmala seeds yielded two pairs of unique racemic pyrroloindole alkaloids, (±)-peganines A-B (1-2); two rare thiazole derivatives, peganumals A-B (3-4); six new ß-carboline alkaloids, pegaharmines F-K (5-10); and 12 known analogues. Their structures, including stereochemistry, were elucidated through spectroscopic analyses, quantum chemistry calculations, and single-crystal X-ray diffraction. Notably, the incorporation of pyrrole and indole moieties in peganines A-B, thiazole fragments in peganumals A-B, and a C-1 α,ß-unsaturated ester motif in pegaharmine F (5) are all rare, and their presence in the genus Peganum were demonstrated for the first time. All isolates were tested for antiproliferative activities against the HL-60, PC-3, and SGC-7901 cancer cell lines, and compounds 9, 11, 12, and 13 exhibited moderate cytotoxicity against HL-60 cancer cell lines with IC50 values in the range of 4.36-9.25 µM.
Subject(s)
Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/isolation & purification , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/isolation & purification , Indole Alkaloids/chemistry , Indole Alkaloids/isolation & purification , Peganum/chemistry , Seeds/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , Carbolines/chemistry , Drug Screening Assays, Antitumor , Drugs, Chinese Herbal/pharmacology , HL-60 Cells , Humans , Indole Alkaloids/pharmacology , Inhibitory Concentration 50 , Molecular Structure , Nuclear Magnetic Resonance, BiomolecularABSTRACT
Two prenylated biflavonoids, podoverines B-C, were isolated from the dried roots and rhizomes of Sinopodophyllum emodi using a Sephadex LH-20 column (SLHC) and high-speed counter-current chromatography (HSCCC). The 95% ethanol extract was partitioned with ethyl acetate in water. Target compounds from the ethyl acetate fraction were further enriched and purified by the combined application of SLHC and HSCCC. n-Hexane-ethyl acetate-methanol-water (3.5:5:3.5:5, v/v) was chosen as the two phase solvent system. The flow rate of mobile phase was optimized at 2.0 mL·min(-1). Finally, under optimized conditions, 13.8 mg of podoverine B and 16.2 mg of podoverine C were obtained from 200 mg of the enriched sample. The purities of podoverines B and C were 98.62% and 99.05%, respectively, as determined by HPLC. For the first time, podoverins B and C were found in the genus Sinopodophyllum. Their structures were determined by spectroscopic methods (HR-ESI-MS, ¹H-NMR, (13)C-NMR, HSQC, HMBC). Their absolute configurations were elucidated by comparison of their experimental and calculated ECD spectra. The cytotoxic activities were evaluated against MCF-7 and HepG2 cell lines. The separation procedures proved to be practical and economical, especially for trace prenylated biflavonoids from traditional Chinese medicine.