High Anticancer Activity Research Articles

Construction of a UDP-Arabinose Regeneration System for Efficient Arabinosylation of Pentacyclic Triterpenoids.

Glycosylation is an important method of modifying natural products and is usually catalyzed by uridine 5'-diphosphate (UDP)-glycosyltransferase. UDP-β-l-arabinose (UDP-Ara) confers specific functions to natural products such as pentacyclic triterpenoids. However, UDP-arabinosyltransferase with high regioselectivity toward pentacyclic triterpenoids has rarely been reported. In addition, UDP-Ara is mainly biosynthesized from UDP-α-d-glucose (UDP-Glc) through several reaction steps, resulting in the high cost of UDP-Ara. Herein, UGT99D1 was systematically characterized for specifically transferring one moiety of arabinose to the C-3 position of typical pentacyclic triterpenoids. Subsequently, 15 enzymes from plants, mammals, and microorganisms were characterized, and a four-enzyme cascade comprising sucrose synthase, UDP-Glc dehydrogenase, UDP-α-d-glucuronic acid decarboxylase, and UDP-Glc 4-epimerase was constructed to transform sucrose into UDP-Ara with UDP recycling. This system was demonstrated to efficiently produce the arabinosylated derivative (Ara-BA) of typical pentacyclic triterpenoid betulinic acid (BA). Finally, the in vitro cytotoxicity tests indicated that Ara-BA showed much higher anticancer activities than BA. The established arabinosylation platform shows the potential to enhance the pharmacological activity of natural products.

Genome-Scale Metabolic Reconstruction, Non-Targeted LC-QTOF-MS Based Metabolomics Data, and Evaluation of Anticancer Activity of Cannabis sativa Leaf Extracts.

Over the past decades, Colombia has suffered complex social problems related to illicit crops, including forced displacement, violence, and environmental damage, among other consequences for vulnerable populations. Considerable effort has been made in the regulation of illicit crops, predominantly Cannabis sativa, leading to advances such as the legalization of medical cannabis and its derivatives, the improvement of crops, and leaving an open window to the development of scientific knowledge to explore alternative uses. It is estimated that C. sativa can produce approximately 750 specialized secondary metabolites. Some of the most relevant due to their anticancer properties, besides cannabinoids, are monoterpenes, sesquiterpenoids, triterpenoids, essential oils, flavonoids, and phenolic compounds. However, despite the increase in scientific research on the subject, it is necessary to study the primary and secondary metabolism of the plant and to identify key pathways that explore its great metabolic potential. For this purpose, a genome-scale metabolic reconstruction of C. sativa is described and contextualized using LC-QTOF-MS metabolic data obtained from the leaf extract from plants grown in the region of Pesca-Boyaca, Colombia under greenhouse conditions at the Clever Leaves facility. A compartmentalized model with 2101 reactions and 1314 metabolites highlights pathways associated with fatty acid biosynthesis, steroids, and amino acids, along with the metabolism of purine, pyrimidine, glucose, starch, and sucrose. Key metabolites were identified through metabolomic data, such as neurine, cannabisativine, cannflavin A, palmitoleic acid, cannabinoids, geranylhydroquinone, and steroids. They were analyzed and integrated into the reconstruction, and their potential applications are discussed. Cytotoxicity assays revealed high anticancer activity against gastric adenocarcinoma (AGS), melanoma cells (A375), and lung carcinoma cells (A549), combined with negligible impact against healthy human skin cells.

A novel water-soluble Cu(II) gluconate complex inhibits cancer cell growth by triggering apoptosis and ferroptosis related mechanisms

Metal copper complexes have attracted extensive attention as potential alternatives to platinum-based anticancer drugs due to their possible different modes of action. Herein, a new copper(II) gluconate complex, namely [Cu(DPQ)(Gluc)]·2H2O (CuGluc, DPQ = pyrazino[2,3-f][1,10]phenanthroline), with good water-solubility and high anticancer activity was synthesized by using D-gluconic acid (Gluc-2H) as an auxiliary ligand. The complex was well characterized by single-crystal X-ray diffraction analysis, elemental analysis, molar conductivity, and Fourier transform infrared spectroscopy (FTIR). The DNA-binding experiments revealed that CuGluc was bound to DNA by intercalation with end-stacking binding. CuGluc could oxidatively cleave DNA, in which 1O2 and H2O2 were involved. In addition, CuGluc was bound to the IIA subdomain of human serum albumin (HSA) through hydrophobic interaction and hydrogen bonding, showing a good affinity for HSA. The complex showed superior anticancer activity toward several cancer cells than cisplatin in vitro. Further studies indicated that CuGluc caused apoptotic cell death in human liver cancer (HepG2) cells through elevated intracellular reactive oxygen species (ROS) levels, mitochondrial dysfunction, cell cycle arrest, and caspase activation. Interestingly, CuGluc also triggered the ferroptosis mechanism through lipid peroxide accumulation and inhibition of glutathione peroxidase 4 (GPX4) activity. More importantly, CuGluc significantly inhibited tumor growth in vivo, which may benefit from the combined effects of apoptosis and ferroptosis. This work provides a promising strategy to develop highly effective antitumor copper complexes by coordinating with the glucose metabolite D-gluconic acid and exploiting the synergistic effects of apoptosis and ferroptosis mechanisms.

<i>In Vitro</i> Antioxidant and Anticancer Activity of Plant Extracts of <i>Tanacetum dolicophyllum</i> (Kitam.) Kitam

The objective of this study was the assessment of antioxidant and cytotoxic activity from Tanacetum dolicophyllum. Soxhlet extraction has been used for preparing plant extracts for which five solvents have been used, i.e., Petroleum ether, benzene, ethyl acetate, ethanol, and water. Proteins, tannins, phlobatannins, amino acids, carbohydrates, alkaloids, steroids, flavonoids, glycosides, cardiac glycosides, and phenols were detected in the sample extracts. In the phytochemical screening of plant extracts, ethyl acetate and ethanolic extract show the best results which further leads to the investigation of antioxidant activity. Ferric-Reducing Antioxidant Power (FRAP), 2,2-diphenyl-2-picrylhydrazyl (DPPH), Reducing Power assay, and Phosphomolybdenum assays were performed to estimate antioxidant activity. In the RPA and TAC, the absorbance of sample extracts increases as there is an increase in concentration. In DPPH and FRAP assay, ethyl acetate showed good results. DPPH assay for ethyl acetate extract with 172.73(µg/ml) and ethanolic extract with 171.07(µg/ml) of IC50 values. The ethyl acetate extract had the highest antioxidant activity. The MTT assay was used to investigate the anticancer activity. The MTT assay discovered that the ethyl acetate extract had the highest anticancer activity against HeLa cells with an IC50 75 µg/mL as compared to a normal cell line J774A. Several compounds present in the extract of ethyl acetate acted as anticancer and contributed to cytotoxic activity.

Efficient pH-Responsive Nano-Drug Delivery System Based on Dynamic Boronic Acid/Ester Transformation.

Chemotherapy is currently one of the most widely used treatments for cancer. However, traditional chemotherapy drugs normally have poor tumor selectivity, leading to insufficient accumulation at the tumor site and high systemic cytotoxicity. To address this issue, we designed and prepared a boronic acid/ester-based pH-responsive nano-drug delivery system that targets the acidic microenvironment of tumors. We synthesized hydrophobic polyesters with multiple pendent phenylboronic acid groups (PBA-PAL) and hydrophilic PEGs terminated with dopamine (mPEG-DA). These two types of polymers formed amphiphilic structures through phenylboronic ester linkages, which self-assembled to form stable PTX-loaded nanoparticles (PTX/PBA NPs) using the nanoprecipitation method. The resulting PTX/PBA NPs demonstrated excellent drug encapsulation efficiency and pH-triggered drug-release capacity. In vitro and in vivo evaluations of the anticancer activity of PTX/PBA NPs showed that they improved the pharmacokinetics of drugs and exhibited high anticancer activity while with low systemic toxicity. This novel phenylboronic acid/ester-based pH-responsive nano-drug delivery system can enhance the therapeutic effect of anticancer drugs and may have high potential for clinical transformations.

Design, Synthesis, and Anticancer Activity of Novel 3,6-Diunsaturated 2,5-Diketopiperazines.

Based on the marine natural products piperafizine B, XR334, and our previously reported compound 4m, fourteen novel 3,6-diunsaturated 2,5-diketopiperazine (2,5-DKP) derivatives (1, 2, 4-6, 8-16), together with two known ones (3 and 7), were designed and synthesized as anticancer agents against the A549 and Hela cell lines. The MTT assay results showed that the derivatives 6, 8-12, and 14 had moderate to good anticancer capacities, with IC50 values ranging from 0.7 to 8.9 μM. Among them, compound 11, with naphthalen-1-ylmethylene and 2-methoxybenzylidene functions at the 3 and 6 positions of 2,5-DKP ring, respectively, displayed good inhibitory activities toward both A549 (IC50 = 1.2 μM) and Hela (IC50 = 0.7 μM) cancer cells. It could also induce apoptosis and obviously block cell cycle progression in the G2/M phases in both cells at 1.0 μM. The electron-withdrawing functions might not be favorable for the derivatives with high anticancer activities. Additionally, compared to piperafizine B and XR334, these semi-N-alkylated derivatives have high liposolubilities (>1.0 mg mL-1). Compound 11 can be further developed, aiming at the discovery of a novel anticancer candidate.

Synthesis, characterizations, antitumor and antimicrobial evaluations of novel Mn(II) and Cu(II) complexes with NNN-tridentate s-Triazine-Schiff base ligand

The new s-triazine hydrazone, 2,4-bis(morpholin-4-yl)-6-[(E)-2-[1-(pyridin-2-yl)ethylidene]hydrazin-1-yl]-1,3,5-triazine, (L) ligand was used to synthesize three new metal complexes. Single-crystal X-ray diffraction analysis indicated that the [MnLCl2] (1) and [CuLCl2].H2O (2) complexes exhibit distorted trigonal bipyramidal coordination geometry with one NNN-tridentate ligand L and two chloride ions. The trigonality index (τ5) was calculated to be 0.332 and 0.235 Ǻ for complexes 1 and 2, respectively. In case of the [CuL(NO3)2] complex (3), the Cu(II) ion is hexa-coordinated with one NNN-tridentate ligand L, one bidentate nitrate and one monodentate nitrate. The supramolecular structure of complexes 1 and 3 was analyzed using Hirshfeld calculations; hydrogen bonding-based interactions have the largest contribution in their molecular packing. The anticancer and antimicrobial activities of the ligand and its complexes (1–3) were examined. The three complexes have higher anticancer activity against colon HCT-116 and lung A-549 cancer cell lines than the free ligand (L). The only exception is complex 1 which has less cytotoxic activity against lung A-549 cancer cell line than L. Interestingly, the Cu(II) complexes have higher cytotoxic activity than the Mn(II) complex, where complex 3 has the best anticancer activity against HCT-116 (IC50 = 7.78 ± 0.27 μg/mL) and A-549 (12.40 ± 0.54 μg/mL) cell lines. In addition, the three complexes showed greater activity against gram-positive bacteria (S. aureus and B. subtilis) than gram-negative bacteria (E. coli and P. vulgaris) where 3 has best antibacterial activity.

Enhancement of cancer-type specific cytotoxicity of natural killer cells via pre-conditioning with cancer cell culture medium

Recently, we have established a method for culturing NK cell-enriched lymphocytes (NKELs) with high anti-cancer activity. These NKELs are found to be highly cytotoxic against various types of cancer cells, and even the extracellular vesicles derived from those NKELs were also demonstrated high anti-cancer activity. In the present study, to find a way to enhance cancer type specificity of these NKELs, the impact of tumor cell-priming of these NKELs on their cytotoxic effect against particular type of cancer cells was investigated. According to our data, when these NKELs were primed with conditioned media (CM) from three different lines of liver cancer cell lines, their cytotoxicity against liver cancer cell lines was significantly increased compared to that of the NKELs without priming. Based on cytokine arrays of the tumor CM and subsequent network analysis, 6 proteins were selected as a potential candidate factors that facilitated the liver cancer cell-specific priming of those NKELs. When these 6 candidate proteins (human recombinant proteins) were separately applied to prime NKELs, 2 out of 6 proteins (DKK1 and THBS1) showed enhanced liver cancer-specific cytotoxicity. With further optimization and elucidation of the underlying mechanisms, it may be possible to prime NKELs for learned cancer type specificity.

Amino acid modified OCMC-g-Suc-β-CD nanohydrogels carrying lapatinib and ginsenoside Rg1 exhibit high anticancer activity in a zebrafish model.

Nanohydrogels show great potential as efficient drug carriers due to their biocompatibility, low toxicity, and high water absorbability. In this paper, we prepared two O-carboxymethylated chitosan (OCMC)-based polymers functionalized with β-cyclodextrin (β-CD) and amino acid. The structures of the polymers were characterized by Fourier Transform Infrared (FTIR) Spectroscopy. Morphological study was carried out on a Transmission Electron Microscope (TEM), and the results indicated that the two polymers had irregular spheroidal structure with some pores distributed on their surface. The average particle diameter was below 500nm, and the zeta potential was above +30mV. The two polymers were further used for preparing nanohydrogels loaded with anticancer drugs lapatinib and ginsenoside Rg1, and the resulting nanohydrogels showed high drug loading efficiency and pH-sensitive (pH = 4.5) drug release behavior. In vitro cytotoxicity investigation revealed that the nanohydrogels exhibited high cytotoxicity against lung cancer (A549) cells. In vivo anticancer investigation was performed in a transgenic Tg(fabp10:rtTA2s-M2; TRE2:EGFP-kras V12) zebrafish model. The results showed that the synthesized nanohydrogels significantly inhibited the expression of EGFP-kras v12 oncogene in zebrafish liver, and the L-arginine modified OCMC-g-Suc-β-CD nanohydrogels loading lapatinib and ginsenoside Rg1 showed the best results.

Integrating Metabolomics and Network Pharmacology to Explore the Mechanism of Tongmai Yangxin Pills in Ameliorating Doxorubicin-Induced Cardiotoxicity.

Doxorubicin (DOX) is a broad-spectrum chemotherapeutic drug used in clinical treatment of malignant tumors. It has a high anticancer activity but also high cardiotoxicity. The aim of this study was to explore the mechanism of Tongmai Yangxin pills (TMYXPs) in ameliorating DOX-induced cardiotoxicity through integrated metabolomics and network pharmacology. In this study, first, an ultrahigh-performance liquid chromatography-quadrupole-time-of-flight/mass spectrometry (UPLC-Q-TOF/MS) metabonomics strategy was established to obtain metabolite information and potential biomarkers were determined after data processing. Second, network pharmacological analysis was used to evaluate the active components, drug-disease targets, and key pathways of TMYXPs to alleviate DOX-induced cardiotoxicity. Targets from the network pharmacology analysis and metabolites from plasma metabolomics were jointly analyzed to select crucial metabolic pathways. Finally, the related proteins were verified by integrating the above results and the possible mechanism of TMYXPs to alleviate DOX-induced cardiotoxicity was studied. After metabolomics data processing, 17 different metabolites were screened, and it was found that TMYXPs played a role in myocardial protection mainly by affecting the tricarboxylic acid (TCA) cycle of myocardial cells. A total of 71 targets and 20 related pathways were screened out with network pharmacological analysis. Based on the combined analysis of 71 targets and different metabolites, TMYXPs probably played a role in myocardial protection through regulating upstream proteins of the insulin signaling pathway, MAPK signaling pathway, and p53 signaling pathway, as well as the regulation of metabolites related to energy metabolism. They then further affected the downstream Bax/Bcl-2-Cyt c-caspase-9 axis, inhibiting the myocardial cell apoptosis signaling pathway. The results of this study may contribute to the clinical application of TMYXPs in DOX-induced cardiotoxicity.

X-ray Crystal Structure-Guided Discovery of Novel Indole Analogues as Colchicine-Binding Site Tubulin Inhibitors with Immune-Potentiating and Antitumor Effects against Melanoma.

A series of novel indole analogues were discovered as colchicine-binding site inhibitors of tubulin. Among them, 3a exhibited the highest antiproliferative activity (average IC50 = 4.5 nM), better than colchicine (IC50 = 65.3 nM). The crystal structure of 3a in complex with tubulin was solved by X-ray crystallography, which explained the improved binding affinity of 3a to tubulin and thus its higher anticancer activity (IC50 = 4.5 nM) than the lead compound 12b (IC50 = 32.5 nM). In vivo, 3a (5 mg/kg) displayed significant antitumor efficacy against B16-F10 melanoma with a TGI of 62.96% and enhanced the antitumor efficacy of a small-molecule PD-1/PD-L1 inhibitor NP19 (TGI = 77.85%). Moreover, 3a potentiated the antitumor immunity of NP19 by activating the tumor immune microenvironment, as demonstrated by the increased tumor-infiltrating lymphocytes (TIL). Collectively, this work shows a successful example of crystal structure-guided discovery of a novel tubulin inhibitor 3a as a potential anticancer and immune-potentiating agent.

Controlled Drug Release from Hydrogel Formulations for The Localised Delivery of Anticancer Agents to Solid Tumors

The hydrogel of the β-sheet self-assembled peptides is one of the powerful vehicles for the drug delivery and other biomedical applications. This class of hydrogel contains both hydrophilic and hydrophobic moieties. Therefore, it plays an integral part in the delivering of the hydrophobic drugs, which considers as a main challenge to overcome when dealing with hydrogels, this is because hydrogels are hydrophilic in nature. Herein, Doxorubicin has been used as a model anticancer agent because it is the most widely known as an anthracycline antibiotic with high anticancer activity. The major challenge with this chemotherapeutic agent its poor aqueous solubility, thus attempts have been made to transform it into hydrogel via hydrophobic interactions. The release of doxorubicin from the hydrogels at the tumour cells, is the vital aim here. Controlling the Dox release has been achievable through monitoring several parameters, such as the gel concentrations, PH, time, and the number of lysine residues. The mechanical properties, secondary structure and the morphology of the peptide hydrogels and Dox hydrogels were also assessed, via using the Rheometer, FTIR and SEM.

Green Synthesis of ZnO and V-Doped ZnO Nanoparticles Using Vinca rosea Plant Leaf for Biomedical Applications.

The present work focused on the synthesis of Vinca rosea leaf extract derived ZnO and vanadium-doped ZnO nanoparticles (V-ZnO NPs). The chemical composition, structural, and morphology of ZnO and vanadium-doped ZnO NPs were examined by FTIR, XRD, and SEM-EDX. The FTIR confirmed the presence of functional groups corresponding to ZnO and vanadium-doped ZnO NPs. SEM-EDX clearly indicated the morphology of synthesised NPs; the hexagonal crystal of NPs was confirmed from XRD. In addition, the cytotoxic effect of ZnO and V-ZnO NPs was estimated against the breast cancer (MCF-7) cell line. From the assay, Vinca rosea (V. rosea) capped ZnO NPs have shown improved cytotoxic activity than that of Vinca rosea capped V-ZnO NPs. ZnO and vanadium-doped ZnO NPs showed the strongest antibacterial activity against Enterococcus, Escherichia coli, Candidaalbicans, and Aspergillus niger. The α-amylase inhibition assays demonstrated antidiabetic activity of synthesised NPs. From the assay test, results obtained Vinca rosea capped ZnO nanoparticles prepared using the green approach showed high effective antioxidant, antidiabetic activity, and anticancer activity than vanadium-doped ZnO NPs.

Exploring the Binding of Natural Compounds to Cancer-Related G-Quadruplex Structures: From 9,10-Dihydrophenanthrenes to Their Dimeric and Glucoside Derivatives.

In-depth studies on the interaction of natural compounds with cancer-related G-quadruplex structures have been undertaken only recently, despite their high potential as anticancer agents, especially due to their well-known and various bioactivities. In this frame, aiming at expanding the repertoire of natural compounds able to selectively recognize G-quadruplexes, and particularly focusing on phenanthrenoids, a mini-library including dimeric (1-3) and glucoside (4-5) analogues of 9,10-dihydrophenanthrenes, a related tetrahydropyrene glucoside (6) along with 9,10-dihydrophenanthrene 7 were investigated here by several biophysical techniques and molecular docking. Compounds 3 and 6 emerged as the most selective G-quadruplex ligands within the investigated series. These compounds proved to mainly target the grooves/flanking residues of the hybrid telomeric and parallel oncogenic G-quadruplex models exploiting hydrophobic, hydrogen bond and π-π interactions, without perturbing the main folds of the G-quadruplex structures. Notably, a binding preference was found for both ligands towards the hybrid telomeric G-quadruplex. Moreover, compounds 3 and 6 proved to be active on different human cancer cells in the low micromolar range. Overall, these compounds emerged as useful ligands able to target G-quadruplex structures, which are of interest as promising starting scaffolds for the design of analogues endowed with high and selective anticancer activity.

Antibacterial and anticancer activity of poly (ɛ-caprolactone)/doxorubicin/Costus arabicus L. root smoke composite nanofibers against lung cancer

The combination of plant-derived agents with chemotherapy drugs could enhance the cytotoxicity of drugs in cancer cells and could improve the biocompatibility of drugs in normal cells. In the present study, the dichloromethane fraction of Costus arabicus L. root smoke (CALRS) and doxorubicin (DOX) have been simultaneously incorporated into the poly (ɛ-caprolactone) (PCL) nanofibers. The synthesized nanofibrous samples were characterized using Fourier-transform infrared spectroscopy, gas chromatography–mass spectrometry, and scanning electron microscopy analysis. The antibacterial, antiparasitic, and anticancer activities of synthesized nanofibers were investigated. The pharmacokinetic models such as Korsmeyer-Peppas, Higuchi, and zero-order were applied to analyze the DOX release mechanism from nanofibers. The continuous release of DOX from PCL/CALRS 5%/DOX 10% nanofibers was occurred during 7 days and the Korsemeyer-Peppas model could describe the DOX release data. The results indicated that the DOX and CALRS dual agents-loaded nanofibers indicated the highest antibacterial activity on both gram-negative Escherichia coli and gram-positive Staphylococcus aureus bacteria and highest anticancer activity on A549 lung cancer cells death compared to pure DOX, CALRS, and DOX loaded-PCL nanofibers.

Triple targeting of mutant EGFRL858R/T790M, COX-2, and 15-LOX: design and synthesis of novel quinazolinone tethered phenyl urea derivatives for anti-inflammatory and anticancer evaluation

We designed and synthesised novel quinazolinone tethered phenyl urea derivatives (6a–p) that triple target the double mutant EGFRL858R/T790M, COX-2, and 15-LOX. Compounds (6e, 6d, 6j, 6m, and 6n) not only had low micromolar IC50 inhibitory activities against the three targets, but they also showed good selectivity for COX-2 over COX-1 and for EGFRL858R/T790M over wild-type EGFR. Except for 6e and 6n, all of the tested compounds inhibited the NO production significantly more potently than celecoxib, diclofenac, and indomethacin. Compounds 6i and 6k reduced ROS levels more effectively than celecoxib and diclofenac. In terms of inhibiting TNF-α production, 6o-treated cells showed TNF-α level, which is ∼10 times lower than celecoxib. Furthermore, 6e and 6j had the highest anticancer activity against the breast cancer cell line BT-459 with growth inhibition percentages of 67.14 and 70.07%, respectively. Docking studies confirm their favoured binding affinity. The proposed compounds could be promising multi-targeted leads.

Cytotoxic effect of Rotheca serrata on cancer cell lines MCF-7 and neuroblastoma SH-SY5Y.

Rotheca serrata (Lamiaceae), a highly medicinal plant is used as an antidote for snakebite and the plant possesses medicinal properties like hepatoprotective, antitussive, antioxidant, anticancer, neuro-protective, used in rheumatoid arthritis and is also a α-glucoside inhibitor. AIM OF THE STUDY: This work aimed to study the anticancerous effect of Rotheca serrata (root and leaf) on cancer cell lines MCF-7 (breast cancer cell line) and Neuroblastoma SH-SY5Y. This investigation was a preliminary one which supported the retrospective and safe use of plants as described in Ayurveda. Dulbecco's Modified Eagle Medium with High Glucose (DMEM-HG) for culturing MCF-7- Human Breast cancer cell line and Minimum essential Medium (MEM)+F12 medium for culturing SH-SY5Y- Homo sapiens bone marrow neuroblast were used. MTT assay measured the cell proliferation rate and conversely, when metabolic events lead to apoptosis or necrosis, the reduction in cell viability. The results indicated that the Methanolic extract of Rotheca serrata (root and leaf) showed high anticancer activity. Different concentrations of plant extracts (25, 50, 100, 200, 400 µg/ml) were used to study the anticancerous activity, amongst which the significant results were obtained for 400 µg/ml concentration (both root & leaf). Effective anticancer activity against MCF - 7 breast cancer cells was shown in methanoilc extracts and were expressed as IC 50 values; in root (IC 50 value = 61.8259 ± 7.428 µg/ml) and in leaf (IC 50 value = 78.1497 ± 6.316 µg/ml). The MTT assay in case of neuroblastoma (SH-SY5Y) cell lines revealed that 400 µg/ml concentration of leaf methanolic extract showed effective inhibition of cancer cells with IC 50 value 37.8462 ± 2.957 µg/ml as compared to IC 50 value of root methanolic extract which was 57.0895 ± 2.351 µg/ml. R. serrata possess anticancer activity against breast cancer cell line (MCF-7) and neuroblastoma (SH-SY 5Y) cell lines. This study may to design plant-based drugs without side effects. Dosage compensation for specific type of cancer needs to be monitored in patients with 1st stage.

Abstract 4901: The role of protein disulfide isomerase and copper in the paraptotic cell death of clinically investigated anticancer thiosemicarbazones

Abstract α-N-Heterocyclic thiosemicarbazones (TSCs) have long been investigated as anticancer compounds. Triapine is one of the best-known TSCs for anticancer therapy and is currently tested in a clinical phase III trial. To further improve the anticancer activity of TSCs, novel derivatives (such as DpC and COTI-2) have been developed and clinically investigated for their activity against solid tumors. These novel TSCs belong to a subclass (incl. the tetra-methylated Triapine derivative, Me2NNMe2) that are terminally disubstituted and demonstrated enhanced anticancer activity in vitro and in vivo. The improved activity of these TSCs correlated with their ability to form stable copper complexes as well as induce paraptosis, a form of programmed but caspase-independent cell death, that is characterized by the formation of vesicles originating from the endoplasmic reticulum (ER). However, the molecular events behind paraptosis induction are still not fully elucidated. Consequently, the aim of this study was to investigate the molecular signaling behind paraptosis as well as the reasons behind the induction of this quite unknown form of cell death by TSCs. A whole-genome gene expression analysis of cells treated with a terminally disubstituted TSC revealed an upregulation of thiol homeostasis-related genes, copper metabolism, and metallothionein genes as well as ER stress response genes. In addition, we found an upregulation of the copper-sensitive and thiol-containing protein disulfide isomerase (PDI), which was confirmed on protein level. In cell-free activity studies, PDI was indeed inhibited by terminally disubstituted TSCs, although only in form of their copper complexes. In agreement, removal of copper ions by specific chelators greatly reduced their anticancer activity as well as paraptotic potential in cell culture. This inhibition was hypothesized to be induced by a general disruption of the thiol redox homeostasis affecting also other responsive proteins and resulting in a more oxidative environment, especially in the ER. In confirmation, thiol-containing small molecules could reduce the compound’s anticancer activity in viability assays. Furthermore, glutathione as well as PDI were detected predominantly in their oxidized form in cells treated with terminally disubstituted TSCs. Consequently, terminally disubstituted TSCs, which are characterized by higher anticancer activity, induce paraptosis due to their high copper complex stability. The formation of the complex results in the interaction with thiol-containing proteins and subsequently in the disruption of the thiol-redox homeostasis, which leads to paraptotic cell death. Overall this work shed light on the paraptotic cell death and will be of interest for future clinical development of anticancer TSCs, as paraptosis is hypothesized to overcome apoptosis-resistance of cancer cells. Citation Format: Sonja Hager, Walter Berger, Christian R. Kowol, Eva A. Enyedy, Petra Heffeter. The role of protein disulfide isomerase and copper in the paraptotic cell death of clinically investigated anticancer thiosemicarbazones. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4901.

Abstract 6787: Vaccinia virus harboring TRAIL (vTRAIL) for in vivo lung cancer therapy

Abstract While oncolytic virotherapy has shown promising results for uncured patients with cancer, its effects on lung cancer remain unclear. Lung cancer is mostly unlikely to respond to conventional anticancer drug, therefore, oncolytic virotherapy can be the best option. Herein, we evaluated the anti-cancer activity of the previously developed three oncolytic vaccinia viruses encoding TNF-related apoptosis-inducing ligand (TRAIL), angiopoietin 1 (Ang1) and both genes, especially on killing lung cancer (LC). In this study, we confirmed that our engineered vaccinia viruses showed overall higher anti-cancer activity on lung cancer cells, compared to WT. Vaccinia virus engineered with both Ang1 and TRAIL (vTRAN) showed the highest oncolytic activity and selectivity in vitro (WT < vTRAIL < vAng1 < vTRAN). However, In vivo LLC1 murine lung cancer syngeneic model showed highest attenuated tumor growth when vTRAIL was treated (WT < vAng1 < vTRAN < vTRAIL), which was in accordance with CD8 high PD1 low expression levels in the tissues. It concludes that engineered vaccinia virus harboring TRAIL (vTRAIL) is the promising therapeutic option for in vivo lung cancer therapy. Citation Format: So Young Yoo. Vaccinia virus harboring TRAIL (vTRAIL) for in vivo lung cancer therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6787.

Development and Characterization of Quercetin-Loaded Delivery Systems for Increasing Its Bioavailability in Cervical Cancer Cells.

Quercetin is a natural flavonoid with high anticancer activity, especially for related-HPV cancers such as cervical cancer. However, quercetin exhibits a reduced aqueous solubility and stability, resulting in a low bioavailability that limits its therapeutic use. In this study, chitosan/sulfonyl-ether-β-cyclodextrin (SBE-β-CD)-conjugated delivery systems have been explored in order to increase quercetin loading capacity, carriage, solubility and consequently bioavailability in cervical cancer cells. SBE-β-CD/quercetin inclusion complexes were tested as well as chitosan/SBE-β-CD/quercetin-conjugated delivery systems, using two types of chitosan differing in molecular weight. Regarding characterization studies, HMW chitosan/SBE-β-CD/quercetin formulations have demonstrated the best results, which are obtaining nanoparticle sizes of 272.07 ± 2.87 nm, a polydispersity index (PdI) of 0.287 ± 0.011, a zeta potential of +38.0 ± 1.34 mV and an encapsulation efficiency of approximately 99.9%. In vitro release studies were also performed for 5 kDa chitosan formulations, indicating a quercetin release of 9.6% and 57.53% at pH 7.4 and 5.8, respectively. IC50 values on HeLa cells indicated an increased cytotoxic effect with HMW chitosan/SBE-β-CD/quercetin delivery systems (43.55 μM), suggesting a remarkable improvement of quercetin bioavailability.