Decrease In IC50 Value Research Articles

Influence of InVitro Digestion on Dipeptidyl Peptidase-IV (DPP-IV) Inhibitory Activity of Plant-Protein Hydrolysates Obtained from Agro-Industrial By-Products.

This study investigates the production of protein hydrolysates with dipeptidyl peptidase-IV (DPP-IV) inhibitory activity from agro-industrial by-products, namely olive seed, sunflower seed, rapeseed, and lupin meals, as well as from two plant protein isolates such as pea and potato. Furthermore, the effect of simulated gastrointestinal digestion on the DPP-IV inhibitory activity of all the hydrolysates was evaluated. Overall, the lowest values of IC50 (1.02 ± 0.09 - 1.24 ± 0.19 mg protein/mL) were observed for the hydrolysates with a high proportion of short-chain [< 1 kDa] peptides (i.e., olive seed, sunflower seed, and lupin) or high content of proline (i.e., rapeseed). Contrarily, the IC50 of the pea and potato hydrolysates was significantly higher (1.50 ± 0.13 - 1.93 ± 0.13 mg protein/mL). In vitro digestion led to an increase in peptides <1 kDa for almost all hydrolysates (except olive and sunflower seed meals), which was noticeable for rapeseed, pea, and potato hydrolysates. Digestion did not significantly modify the DPP-IV inhibitory activity of olive, sunflower, rapeseed, and potato hydrolysates, whereas a significant decrease in IC50 value was obtained for pea hydrolysate and a significant increase in IC50 was obtained for lupin hydrolysate. Thus, this work shows the potential of agro-industrial by-products for the production of protein hydrolysates exhibiting DPP-IV inhibition.

Synergistic anticancer effect of polymersome‐encapsulated curcumin and cytarabine: A spectroscopic and theoretical approach

AbstractCytarabine, also known as cytosine arabinoside is an anticancer drug, and it is widely used to treat acute myeloid leukemia (AML). But, clinical efficacy of highly hydrophilic cytarabine is often limited due to its rapid degradation and high biodistribution. Hence, this study aims to enhance the anticancer effect of cytarabine with the help of a naturally occurring polyphenol, curcumin. The interaction between cytarabine and curcumin was established by different spectroscopic techniques like UV–Visible, fluorescence, and Fourier transform infrared (FT‐IR). Molecular interactions were also studied using isothermal calorimetric titrations (ITC) followed by theoretical docking. The involvement of the amine group of cytarabine with the interaction of curcumin has been supported by FT‐IR and docking study. However, the major challenge with curcumin is that, in spite of its several medicinal benefits, poor solubility and bioavailability restricts its potential clinical applications. Hence, polymersome‐encapsulated curcumin (PEC) was prepared for the intravenous application. A two‐fold decrease in IC50 value of cytarabine against HeLa cell line was found in presence of encapsulated curcumin. This was justified on the basis of observed reactive oxygen species enhancement in serum in presence of cytarabine and PEC together, determined using absorption spectroscopy. The enhanced affinity of cytarabine toward DNA‐curcumin complex as compared to free DNA, supported by theoretical study, also validates the increased cytotoxicity of cytarabine in presence of PEC.

Green Synthesis of Silver Nanoparticle Using Black Mulberry and Characterization, Phytochemical, and Bioactivity.

Synthesis of silver nanoparticles (AgNPs) using plant extracts has been proposed as a more advantageous and environmentally friendly alternative compared to existing physical/chemical methods. In this study, AgNPs were synthesized from silver nitrate using black mulberry (BM) extract. The biosynthesized AgNPs were characterized through an UV-visible spectrometer, X-ray diffraction, and transmission electron microscopy. Additionally, BM-AgNPs were subjected to antioxidant, antibacterial, anti-inflammatory, and anticancer activities. AgNPs biosynthesized from BM extract were dark brown in color and showed a strong peak at 437 nm, confirming that AgNPs were successfully synthesized. The size of AgNPs was 170.17 ± 12.65 nm, the polydispersity index was 0.281 ± 0.07, and the zeta potential value was -56.6 ± 0.56 mV, indicating that the particles were stable. The higher total phenol, flavonoid, and anthocyanin content of BM-AgNPs compared to BM extract indicates that the particles contain multiple active substances due to the formation of AgNPs. The DPPH and ABTS assays showed decreased IC50 values compared to BM extract, demonstrating improved antioxidant activity. AgNPs inhibited the growth of S. aureus and E. coli at 600 μg/mL, with minimum bactericidal concentrations determined to be 1000 and 1200 μg/mL, respectively. The anti-inflammatory activity was 64.28% at a BM-AgNPs concentration of 250 μg/mL. As the concentration increased, the difference from the standard decreased, indicating the inhibitory effect of AgNPs on bovine serum albumin denaturation. The viability of MCF-7 cells treated with BM-AgNPs was found to be significantly lower than that of cells treated with BM extract. The IC50 value of BM-AgNPs was determined to be 96.9 μg/mL. This study showed that BM-AgNPs have the potential to be used in the pharmaceutical industry as antioxidant, antibacterial, anti-inflammatory, and anticancer agents.

Identification of SMC2 and SMC4 as prognostic markers in breast cancer through bioinformatics analysis.

Breast cancer (BRCA) is one of the most common malignant tumors. The structural maintenance of chromosome (SMC) gene family has been shown to play an important role in human cancers. However, the role of SMC families in BRCA is unclear. This study aimed to explore the role and potential clinical value of whole SMCs in BRCA. TIMER and UALCAN database were used to analysis the expression level. Genetic variations were analyzed by cBioPortal. Promoter methylation and protein level were analyzed by UCLCAN. GO and KEGG were analyzed by Metascape database. Prognostic value of SMCs was analyzed by Kaplan-Meier and multivariate cox regression analyses. Immune infiltration analysis was conducted by CIBERSORT. Immunotherapy outcome prediction was conducted by Cancer Immunome Atlas. Targeted drug therapy outcome prediction was taken by GDSC and R language. The cell viability was tested by CCK8 and migration was tested by wound healing assay. Xenograft model was used to investigate the in vivo role of SMC2. Expression levels of SMC1A, SMC2, SMC4, SMC5 and SMC6 mRNA were increased in BRCA tissues, and negatively correlated with promoter methylation. Overexpression of SMC2 and SMC4 was negatively correlated with survival. Function of SMCs family regulatory genes was mainly related to ATPase activity. Expression of most SMCs was negatively correlated with immunotherapy and drug therapy outcomes. Interfere SMC2 and SMC4 decreased IC50 values of 5-fluorouracil and oxaliplatin and inhibited the migration of MCF7 cells. Tumor growth and weights were significantly decreased in si-SMC2 groups. Combined bioinformatics and clinical specimen analysis verified SMC2 and SMC4 as independent prognostic factors in BRCA, suggesting their significance for the diagnosis and treatment of BRCA.

Abstract 5866: TESC mediates resistance to lenvatinib in hepatocellular carcinoma by regulating cell autophagy

Abstract Objective: To investigate the mechanism by which TESC (Tescalcin) induces lenvatinib resistance in hepatocellular carcinoma (HCC) through autophagy mediation. Methods:1) Utilized established liver cancer organoids from over 70 patients for lenvatinib drug sensitivity screening and transcriptome sequencing to identify lenvatinib-resistant genes. 2) Constructed stable TESC overexpression and downregulation liver cancer cell lines. Employed CCK8 assays to plot IC50 drug sensitivity curves, elucidating changes in lenvatinib sensitivity in liver cancer cells after TESC overexpression or downregulation. 3) Used TESC-stable cells to establish subcutaneous tumor models and intrahepatic tumor models in nude mice. After tumor formation, administered lenvatinib orally to observe whether TESC affects liver cancer sensitivity to lenvatinib in vivo. 4) Conducted Western blot (WB) analysis to assess the expression of autophagy-related molecules (LC3B II/I, P62, and Beclin1 proteins) in TESC downregulated or overexpressed stable cell lines and their control cell lines. Utilized the LC3-GFP-mRFP fluorescence dual-labeling lentivirus system and laser confocal microscopy to examine autophagic flux. 5)Treated TESC downregulated cells with the autophagy inhibitor 3-MA. Conducted WB to assess the expression levels of autophagy-related proteins and plotted drug sensitivity curves using IC50 experiments to evaluate the impact of 3-MA on reversing liver cancer cell sensitivity to lenvatinib. Results: Compared to the control group, HCC cells overexpressing TESC showed an increased IC50 value and reduced sensitivity to lenvatinib, while TESC downregulation resulted in a decreased IC50 value and decreased sensitivity to lenvatinib. Subcutaneous tumor experiments in nude mice indicated that, compared to the control group, tumors with reduced TESC expression were more sensitive to lenvatinib. Additionally, in HCC cells overexpressing TESC, protein expression levels of LC3B II/I and Beclin1 significantly decreased, while P62 expression increased. Conversely, TESC downregulation significantly increased protein expression levels of LC3B II/I and Beclin1, while P62 expression decreased. Furthermore, results from the LC3-GFP-mRFP autophagy dual-labeling lentivirus system showed a significant reduction in fluorescence spots in the TESC overexpression group compared to the control group, indicating weakened autophagic flux. Treatment of TESC downregulated liver cancer cells with 3-MA resulted in an increased IC50 value compared to the control group. Conclusion: TESC expression levels are associated with the sensitivity of liver cancer cells to lenvatinib, possibly by inhibiting autophagy and affecting the sensitivity of liver cancer cells to lenvatinib. Citation Format: Hao Zhuang, Tingting Zhang, Jiaqi Chen. TESC mediates resistance to lenvatinib in hepatocellular carcinoma by regulating cell autophagy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5866.

Abstract 2035: A Benzenesulfornamide with potential to improve chemoresponse of ovarian cancer through activation of AMPK and immunogenic cell death

Abstract Introduction: Ovarian cancer is a leading cause of death among women globally often characterized by poor prognosis and aggressive tumor growth. The therapeutic outcomes of ovarian cancer patients are limited by the development of acquired chemoresistance and the lack of targeted therapies. This study aimed to understand the mechanism in which a benzenesulfornamide named Y3 enhances the efficacy of chemotherapeutic agents. Y3 was previously identified by our team as an activator of AMPK with the activity to induce immunogenic cell death of ovarian cancer cells. Material and Methods: Patient-derived ovarian cancer cell lines were pretreated with Y3 for 3 h before treatment of carboplatin or paclitaxel. Celltiter Glo assay was performed to determine the impact of Y3 pretreatment on the IC50 values of carboplatin or paclitaxel after 48 h. Apoptosis was evaluated using Annexin VI/PI staining and Caspase 3 activity assays. The role of AMPK pathway was assessed using AMPK siRNAs and western blot. The potential of Y3 to enhance immunogenic cell death induced by carboplatin or paclitaxel was evaluated in a syngeneic mouse model of ovarian cancer using a vaccination assay. The control group mice were vaccinated by subcutaneously injecting mouse ovarian cancer cells treated by carboplatin in vitro for 24 h into the left flank. The test group mice were vaccinated by injecting Y3-carboplatin co-treated cells. One week after vaccination, untreated live cancer cells were injected into the right flank of both groups. Tumor growth was monitored. Results: Ovarian cancer cell lines pretreated with Y3 showed decreased IC50 values for carboplatin and paclitaxel treatments. The apoptotic cell populations stained by Annexin VI were increased in the Y3 pretreated in comparisons with the single treatment groups. Caspase-3 activity validated the results of Annexin VI/PI staining assay. AMPK knockdown by siRNAs effectively reduced the levels of phospho-AMPK that was induced by Y3, which also dampened the Y3-promoted chemotherapeutic effects on ovarian cancer cells. In the in vivo vaccination assay, tumor growth in the right flank was suppressed in the mice vaccinated with Y3-carboplatin co-treated cells to an extend that is more significant than the control group. Conclusion: Y3 pretreatment increased the sensitivity of ovarian cancer cell lines to carboplatin and paclitaxel. The effects of Y3 on ovarian cancer cells depend on the activation of AMPK. Y3 enhances the immunogenicity of carboplatin-treated ovarian cancer cells. Citation Format: Juhyun Kim, Nia Montero, Shefali Sharma, Miranda Mansolf, Kevin Yang, Tobias Hartwich, Yang Yang-Hartwich. A Benzenesulfornamide with potential to improve chemoresponse of ovarian cancer through activation of AMPK and immunogenic cell death [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2035.

Copper-based metal-organic frameworks (BDC-Cu MOFs) as supporters for α-amylase: Stability, reusability, and antioxidant potential

Copper-based metal-organic frameworks (BDC-Cu MOFs) were synthesized via a casting approach using 1,4-benzene dicarboxylic (BDC) as organic ligand and their properties characterized. The obtained materials were then utilized to immobilize the α-amylase enzyme. The chemical composition and functional components of the synthesized support (BDC-Cu MOFs) were investigated with Fourier transform infrared spectroscopy (FTIR), the surface morphology was determined with scanning electron microscopy (SEM), and the elemental composition was established with energy dispersive X-ray (EDX) analyses. X-ray diffraction (XRD) was employed to analyze the crystallinity of the synthesized DBC-Cu MOFs. The zeta potentials of DBC-Cu MOFs and DBC-Cu MOFs@α-amylase were determined. The immobilized α-amylase demonstrated improved catalytic activity and reusability compared to the free form. Covalent attachment of the α-amylase to BDC-Cu provided an immobilization yield (IY%) of 81% and an activity yield (AY%) of 89%. The immobilized α-amylase showed high catalytic activity and 81% retention even after ten cycles. Storage at 4 °C for eight weeks resulted in a 78% activity retention rate for DBC-Cu MOFs@α-amylase and 49% retention for the free α-amylase. The optimum activity occurred at 60 °C for the immobilized form, whereas the free form showed optimal activity at 50 °C. The free and immobilized α-amylase demonstrated peak catalytic activities at pH 6.0. The maximum reaction velocities (Vmax) values were 0.61 U/mg of protein for free α-amylase and 0.37 U/mg of protein for BDC-Cu MOFs@α-amylase, while the Michaelis‒Menten affinity constants (Km) value was lower for the immobilized form (5.46 mM) than for the free form (11.67 mM). Treatments of maize flour and finger millet samples with free and immobilized α-amylase resulted in increased total phenolic contents. The enhanced antioxidant activities of the treated samples were demonstrated with decreased IC50 values in ABTS and DPPH assays. Overall, immobilization of α-amylase on BDC-Cu MOFs provided improved stability and catalytic activity and enhanced the antioxidant potentials of maize flour and finger millet.

Food emulsifiers increase toxicity of food contaminants in three human GI tract cell lines

Food products simultaneously containing both food contaminants and emulsifiers are common in baked products, coffee and chocolate. Little is known regarding how food contaminants and emulsifiers interact and alter toxicity. Recent studies have shown that while emulsifiers themselves have little toxicity, they could cause changes in the gut microenvironment and lead to issues such as increased uptake of allergens. This study examined toxic effect of two common process contaminants acrylamide (AA) and benzo [a]pyrene (BAP) combined with food emulsifiers polyoxyethylene sorbitan monooleate (TW) or glycerol monostearate (G). In liver cell line HepG2 and gastrointestinal cell lines HIEC6 and Caco-2, toxicities of AA and BAP were increased by TW but not by G as indicated by decrease in IC50 values. Addition of TW also exacerbated gene expression changes caused by AA or BAP. Cellular uptake and cell membrane permeability were enhanced by TW but not by G, but tight junction proteins of Caco-2 monolayer was impacted by both emulsifiers. These results suggested that TW could increase toxicity of AA and BAP through increasing cell permeability thus chemical uptake and potentially through other interactions. The study is to draw the attention of regulators on the potential synergistic interaction of co-occurring chemicals in food.

Synthesis and Biological Evaluation of New Benzochromenopyrimidines for the Therapy of Colon and Lung Cancer

AbstractHerein we report the synthesis of ten newly substituted 3‐phenyl‐5‐aryl‐3,5‐dihydro‐4H‐benzo[6,7]chromeno[2,3‐d]pyrimidin‐4,6,11‐triones (“benzochromenopyrimidines”) in a one‐step reaction, by reacting ethyl 2‐amino‐4‐aryl‐5,10‐dioxo‐5,10‐dihydro‐4H‐benzo[g]chromene‐3‐carboxylates with aniline, and triethyl orthoformate, under acidic catalyst and solvent‐free conditions. These benzochromenopyrimidines were submitted to biological evaluation for in vitro anti‐cancer activity, several derivatives showing higher activity than commonly used chemotherapeutic agents such as oxaliplatin, 5‐fluorouracil, and paclitaxel. In particular, the 5‐(2‐Methoxyphenyl)‐3‐phenyl‐3,5‐dihydro‐4H‐benzo[6,7]chromeno[2,3‐d]pyrimidine‐4,6,11‐trione was identified as the most potent inhibitor among all the new ligands, showing significant cytotoxic activity against all the cancer cell lines tested [human colon cancer cell lines, LoVo and HCT‐116; human lung cancer cells (A549 line)], as evidenced by the significant decrease in IC50 values, while maintaining low cytotoxicity against normal cells. These promising results suggest that this compound deserves further investigation as a potential hit compound for the development of a selective anticancer drug in cancer therapy.

Synthesis and in vitro assessment of gold nanoparticles conjugated with extracts, sterols and pure compounds derived from marine sponges from the Indian and Pacific Oceans.

Gold nanoparticles (AuNPs) exhibit different physical properties compared to small molecules, bulk materials and other nanoparticles. Their synthesis using plant extracts, particularly polyflavonoids as phytoreductants, for the conversion of Au(iii) into Au(0) has been reported. In this study, AuNPs were synthesized with extracts, sterols and pure compounds derived from marine sponges using gold(iii) chloride trihydrate. Extracts, hexane (JDH) and ethyl acetate (JDE), sterols (JC-2) and jaspamide were obtained from Jaspis diastra. Pure compounds, namely, contignasterol, ansellone A, motuporamines A and MN100 (a synthetic analog of pelorol), were also used. JC-2 was characterized using NMR and GC-MS, and the major constituent was determined to be β-sitosterol. β-Sitosterol has shown great promise as an anti-cancer molecule, but its poor aqueous solubility and bioavailability coupled with low targeting efficacy limit its therapeutic efficacy. Transmission electron microscopy (TEM) images revealed the formation of spherical AuNPs conjugated with JDH, JDE, JC-2, ansellone and contignasterol with average diameters of 21.1 ± 3.0 nm, 20.7 ± 2.1 nm, 26.2 ± 1.2 nm, 33.3 ± 5.1 nm and 30.8 ± 5.5 nm, respectively. No particle formation was seen with motuporamines A and MN100. Zeta potential values indicated that AuNPs-JC-2 was more stable than AuNPs-JDE, AuNPs-JDH and AuNPs-ansellone. Based on IC50 values, the cytotoxicity of AuNPs-JDH increased in A172, TERA, HeLa and HepG2 cells but showed similar activity in HaCaT cells compared to JDH. The cytotoxicity of AuNPs-JDE decreased in A172 and HaCaT cells but increased in TERA1, HeLa and HepG2 cells compared to JDE. AuNPs-JC-2 showed enhanced cytotoxicity with a decrease in IC50 values from 3.37 ± 0.19 μg mL-1 to 0.52 ± 0.09 μg mL-1 in A172 and from 2.28 ± 0.20 μg mL-1 to 0.78 ± 0.28 μg mL-1 in TERA1 compared to JC-2. The synergistic action of sterols in AuNPs-JC-2 seemed to favour enhanced anti-cancer activity. The presence of sterols increased the ability of transforming Au(iii) into Au(0) to form AuNPs and further enhancing cellular uptake and, thus, anti-cancer activity. AuNPs-contignasterol displayed lower activity than contignasterol in the A172 cell line. No significant difference in activity was observed with AuNPs-ansellone A in the A172 and HaCaT cell lines compared to ansellone A.

Comparison of Antioxidant Activity of Crude Extract And Gel Preparation From Alstonia Scholaris L. Leaf Extract

Alstonia scholaris L. contains several compounds, which has been proven to be potential as skin protection by formulating in gel preparation. The aims of this study were to obtained the antioxidant activity of crude extract from leaf part of Alstonia scholaris L. and gel preparation contain Alstonia scholaris L. leaf extract. Extract was prepared using destilled methanol by maceration method. The gel preparation was developed with extract (1% w/v) in carbomer base (1,67% w/v). The chemical compound was evaluated qualitatively used some reagent. The antioxidant assays performed by DPPH (2,2-diphenyl-1-picrylhydrazyl) method. The result shown that methanolic extract of Alstonia scholaris leaf was found had flavonoids, tannins, phenols, alkaloids, saponins, and steroids compound. The methanolic crude extract and gel preparation had strong antioxidant activity with IC50 value was found to be 69,502 ppm and 86,832 ppm, respectively. From the result we conclude that the methanolic crude extract of Alstonia scholaris leaf sole has strong scavenging free radical activity even formulated in gel preparation but has decreased IC50 value.

Production of high-antioxidant yoghurt using phycocyanin from microalgae Spirulina sp

Yoghurt is a functional food product widely used to improve the digestive system in the body. Yoghurt fermentation usually uses lactic acid bacteria from the Lactobacillus bulgaricus, Streptococcus thermophilus, and Lactobacillus casei. Phycocyanin has been used as a natural dye for food, cosmetics, and medicine. Phycocyanin is a complex protein that can be an anticancer, antioxidant, and immunity booster. In this study, the effect of the number of additions of phycocyanin, storage time, and time of addition of phycocyanin to the antioxidant activity, protein, and organoleptic properties of the yoghurt produced. In this study, cow milk was pasteurized and fermented to become a yoghurt. Afterwards, phycocyanine was added to the yoghurt before being stored and analyzed. Spectrophotometric analysis was utilized to determine the antioxidant activity and protein content. An organoleptic test using a Likert scale was conducted to determine the product's suitability to consumer tastes. Increased phycocyanin concentrations were added (0, 0.25, 0.5, and 0.75 wt. %), resulting in a decrease in IC50 value of 8855.53, 5843.371, and 4147.548 ppm, which shows an increase in antioxidant activity. At the level of consumer preference in terms of taste, the most preferred is a concentration of 0.5 wt. %, while in terms of colour and aroma, at a concentration of 0.25 wt. %. While the longer the storage time (1, 3, and 5 days), the increase in IC50 value of 2777.111, 5179.547, and 5916.884 ppm, and there was a decrease in protein content in the sample. For the level of consumer preferences in terms of taste and aroma, the most preferred is variable with a storage time of 1 day, and in terms of colour, the most preferred at five days storage. The IC50 value in the variable with the addition of phycocyanin after fermentation is less than the addition after fermentation, which is 5351.865 ± 1606 and 3897.162 ± 1678 ppm. The protein content is higher when adding phycocyanin before fermentation. While yoghurt with the addition phycocyanin before fermentation showed higer consumer preference in terms of taste and aroma. In contrast, the addition of phycocyanin after fermentation is preferred in terms of colour.

Carborane-Based ABCG2-Inhibitors Sensitize ABC-(Over)Expressing Cancer Cell Lines for Doxorubicin and Cisplatin.

The ABCG2 transporter protein, as part of several known mechanisms involved in multidrug resistance, has the ability to transport a broad spectrum of substrates out of the cell and is, therefore, considered as a potential target to improve cancer therapies or as an approach to combat drug resistance in cancer. We have previously reported carborane-functionalized quinazoline derivatives as potent inhibitors of human ABCG2 which effectively reversed breast cancer resistance protein (BCRP)-mediated mitoxantrone resistance. In this work, we present the evaluation of our most promising carboranyl BCRP inhibitors regarding their toxicity towards ABCG2-expressing cancer cell lines (MCF-7, doxorubicin-resistant MCF-7 or MCF-7 Doxo, HT29, and SW480) and, consequently, with the co-administration of an inhibitor and therapeutic agent, their ability to increase the efficacy of therapeutics with the successful inhibition of ABCG2. The results obtained revealed synergistic effects of several inhibitors in combination with doxorubicin or cisplatin. Compounds DMQCa, DMQCc, and DMQCd showed a decrease in IC50 value in ABCB1- and ABCG2-expressing SW480 cells, suggesting a possible targeting of both transporters. In an HT29 cell line, with the highest expression of ABCG2 among the tested cell lines, using co-treatment of doxorubicin and DMQCd, the effective inhibitory concentration of the antineoplastic agent could be reduced by half. Interestingly, co-treatment of compound QCe with cisplatin, which is not an ABCG2 substrate, showed synergistic effects in MCF-7 Doxo and HT29 cells (IC50 values halved or reduced by 20%, respectively). However, a literature-known upregulation of cisplatin-effluxing ABC transporters and their effective inhibition by the carborane derivatives emerges as a possible reason.

MicroRNA-367-3p directly targets RAB23 and inhibits proliferation, migration and invasion of bladder cancer cells and increases cisplatin sensitivity

ObjectivesThis study investigated the biological role of miR-367-3p upregulation in bladder cancer and verified the mutual relation between miR-367-3p and RAB23.Materials and methodsExpression levels of miR-367-3p were determined by RT–qPCR in bladder cancer cell lines and human bladder cancer tissues. The effects of miR-367-3p on proliferation, migration and invasion were evaluated by cell colony formation assays, wound healing assays and trans-well assays, respectively. The effects of miR-367-3p and RAB23 on cisplatin sensitivity of bladder cancer cells were assessed by CCK-8 assay. The expression of its target-RAB23 was determined by western blotting in T24, 5637. Plasmids used in dual-luciferase assays were constructed to confirm the action of miR-367-3p on downstream target-RAB23 in T24 cells. And also, the role of miR-367-3p in tumorigenesis was also confirmed in nude mouse models.ResultsThe downregulation of miR-367-3p was observed in human bladder cancer tissues. MiR-367-3p downregulation positively correlated with tumor stage and tumor grade. MiR-367-3p overexpression in T24, 5637 cells suppressed the proliferation, migration, and invasion of bladder cancer cells in vitro while decreasing IC50 values under T24 and 5637 cisplatin treatment conditions. RAB23 was shown to be upregulated in bladder cancer tissues and cell lines. MiR-367-3p directly bound to the 3′ UTR of RAB23 in T24 cells. RAB23 was potentially accounted for the aforementioned functions of miR-367-3p. Tumor formation experiments in nude mouse models confirmed that overexpression of miR-367-3p could inhibit tumor growth and invasion in vivo.ConclusionsmiR-367-3p acts as a tumor suppressor in bladder cancer by downregulating RAB23 signaling. We conjecture that miR-367-3p-mediated downregulation of RAB23 expression may be a new therapeutic strategy for bladder cancer treatment.

Evaluating the In Vitro Activity and Safety of Modified LfcinB Peptides as Potential Colon Anticancer Agents: Cell Line Studies and Insect-Based Toxicity Assessments.

Anticancer peptides are increasingly being considered as alternative treatments for cancer due to their potency, selectivity, and low toxicity. Previously, the peptide LfcinB (21-25)Pal showed in vitro anticancer effects against the Caco-2 colon cancer cell line (half-maximal inhibitory concentration (IC50): 86 μM). In this study, we developed modifications to the peptide sequence to increase its anticancer activity. Sequence modifications were made such as the inclusion of amino hexanoic acid (Ahx), N-terminal biotinylation, acetylation, and substitutions of Orn for Arg and/or d-Arg by l-Arg. The molecules were synthesized using manual solid-phase peptide synthesis (SPPS), and their synthetic feasibility (SAScore) ranged from 6.2 to 7.6. The chromatographic purities of the synthesized peptides were greater than 89%. We found that Ahx-RWQWRWQWR and RWQWRWQW-Orn showed activity against both Caco-2 and HT-29 cell lines and decreased IC50 values by approx. 50% in Caco-2 cells (IC50: 40 μM) when compared to the parent peptide RWQWRWQWR. Moreover, the modified peptides demonstrated lower hemolytic effects, with values <10% at 200 μg/mL. Toxicity was assessed using the Galleria mellonella model and the half-maximal lethal dose (LD50) for the best peptides was >100 mg/kg, indicating that their toxicity is classified as moderately toxic or lower. In contrast, cisplatin showed an LD50 of 13 mg/Kg. The designed anticancer peptides presented good in vitro activity and low toxicity, making them promising molecules for future drug development studies.

Cellular Efficacy of Fattigated Nanoparticles and Real-Time ROS Occurrence Using Microfluidic Hepatocarcinoma Chip System: Effect of Anticancer Drug Solubility and Shear Stress.

The objective of this study was to evaluate the effectiveness of organ-on-chip system investigating simultaneous cellular efficacy and real-time reactive oxygen species (ROS) occurrence of anticancer drug-loaded nanoparticles (NPs) using hepatocarcinoma cells (HepG2) chip system under static and hepatomimicking shear stress conditions (5 dyne/cm2). Then, the role of hepatomimetic shear stress exposed to HepG2 and drug solubility were compared. The highly soluble doxorubicin (DOX) and poorly soluble paclitaxel (PTX) were chosen. Fattigated NPs (AONs) were formed via self-assembly of amphiphilic albumin (HSA)-oleic acid conjugate (AOC). Then, drug-loaded AONs (DOX-AON or PTX-AON) were exposed to a serum-free HepG2 medium at 37 °C and 5% carbon dioxide for 24 h using a real-time ROS sensor chip-based microfluidic system. The cellular efficacy and simultaneous ROS occurrence of free drugs and drug-loaded AONs were compared. The cellular efficacy of drug-loaded AONs varied in a dose-dependent manner and were consistently correlated with real-time of ROS occurrence. Drug-loaded AONs increased the intracellular fluorescence intensity and decreased the cellular efficacy compared to free drugs under dynamic conditions. The half-maximal inhibitory concentration (IC50) values of free DOX (13.4 μg/mL) and PTX (54.44 μg/mL) under static conditions decreased to 11.79 and 38.43 μg/mL, respectively, under dynamic conditions. Furthermore, DOX- and PTX-AONs showed highly decreased IC50 values of 5.613 and 21.86 μg/mL, respectively, as compared to free drugs under dynamic conditions. It was evident that cellular efficacy and real-time ROS occurrence were well-correlated and highly dependent on the drug-loaded nanostructure, drug solubility and physiological shear stress.

Enantiomeric Virus-Inspired Oncolytic Particles for Efficient Antitumor Immunotherapy.

Synthesizing biomimetic systems with stereospecific architectures and advanced bioactivity remains an enormous challenge in modern science. To fundamentally eliminate biosafety issues of natural oncolytic viruses, the development of synthetic virus-inspired particles with high oncolytic activity is urgently needed for clinical antitumor treatments. Here, we describe the design and synthesis of enantiomeric virus-inspired particles for efficient oncolytic therapy from homochiral building blocks to stereospecific supramolecular constructions. The L-virus-inspired oncolytic particles (L-VOPs) and D-VOPs possess similar biomimetic nanostructures but mirror-imaged enantiomeric forms. It is important that both L-VOPs and D-VOPs successfully mimic the pharmacological activity of oncolytic viruses, including direct tumor lysis and antitumor immune activation. D-VOPs provide quite better oncolytic efficacy than that of clinical-grade oncolytic agents (LTX-315, IC50 = 53.00 μg mL-1) with more than 5-fold decrease in IC50 value (10.93 μg mL-1) and close to 100% tumor suppression (98.79%) against 4T1 tumor-bearing mice, attributed to the chirality-dependent tumor recognition, interaction, antidegradation, and immunotherapy. This work provides a strategy for the synthesis of stereospecific biomimetic material systems as well as develops an advanced candidate for biomimetic oncolytic agents without biosafety risks.

Tyrosinase-Catalyzed Peptide Macrocyclization for mRNA Display.

mRNA display of macrocyclic peptides has proven itself to be a powerful technique to discover high-affinity ligands for a protein target. However, only a limited number of cyclization chemistries are known to be compatible with mRNA display. Tyrosinase is a copper-dependent oxidase that oxidizes tyrosine phenol to an electrophilic o-quinone, which is readily attacked by cysteine thiol. Here we show that peptides containing tyrosine and cysteine are rapidly cyclized upon tyrosinase treatment. Characterization of the cyclization reveals it to be widely applicable to multiple macrocycle sizes and scaffolds. We combine tyrosinase-mediated cyclization with mRNA display to discover new macrocyclic ligands targeting melanoma-associated antigen A4 (MAGE-A4). These macrocycles potently inhibit the MAGE-A4 binding axis with nanomolar IC50 values. Importantly, macrocyclic ligands show clear advantage over noncyclized analogues with ∼40-fold or greater decrease in IC50 values.

Enhancement of in-vitro anti-oral cancer activities of silymarin using dispersion of nanostructured lipid carrier in mucoadhesive in-situ gel

Silymarin (SME) shows multiple therapeutic actions against several cancers, however, low aqueous solubility and poor bioavailability issues restrict its clinical use. In this study, SME was loaded in nanostructured lipid carriers (NLCs) and further incorporated in mucoadhesive in-situ gel (SME-NLCs-Plx/CP-ISG) for localized treatment of oral cancer. Using a 33 Box-Behnken design (BBD), an optimized SME-NLC formula was developed with the ratios of solid lipids, surfactant concentration, and sonication time as independent variables, while particle size (PS), polydispersity index (PDI), and % encapsulation efficiency (EE) as dependent variables, resulting in 315.5 ± 0.1 nm PS, 0.341 ± 0.01 PDI, and 71.05 ± 0.05 % EE. Structural studies confirmed the formation of SME-NLCs. SME-NLCs incorporated in-situ gel demonstrated a sustained release for SME, indicating enhanced retention on the buccal mucosal membrane. The in-situ gel containing SME-NLCs showed a marked decrease in IC50 value (24.90 ± 0.45 µM) than SME-NLCs (28.40 ± 0.89 µM) and plain SME (36.60 ± 0.26 µM). The studies demonstrated that Reactive oxygen species (ROS) generation potential and SME-NLCs-Plx/CP-ISG induced apoptosis at Sub-G0 phase owing to higher penetration of SME-NLCs led to higher inhibition against human KB oral cancer cells. Therefore, SME-NLCs-Plx/CP-ISG can be the alternative to chemotherapy and surgery with site-specific delivery of SME to oral cancer patients.

Radical scavenging abilities of L-tyrosine and L-DOPA Schiff bases and their fluorescence binding studies and molecular docking interactions with bovine serum albumin

Phenolic compounds are known for their abilities to scavenge and eliminate reactive oxygen species (ROS) which may lead to oxidative stress. Bearing in mind that Schiff bases are biologically active compounds, a series of Schiff bases derived from the monophenolic l-tyrosine and diphenolic l-DOPA were synthesized and characterized by spectral and elemental analysis. The structure of the l-DOPA naphthyl Schiff base was confirmed and characterized by X-ray. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical assays were used in assessing the effectiveness of these compounds as radical scavengers. l-DOPA derivatives were better radical scavengers with lower IC50 compared to l-tyrosine derivatives. To understand the distribution of these compounds in the blood plasma, their interactions with bovine serum albumin (BSA) were monitored using fluorescent emission spectroscopy where they were found to quench the BSA intrinsic fluorescence by combined static and dynamic mechanism. The fluorescence and FT-IR data showed that conformational changes occurred in BSA after interaction with the Schiff bases. The binding constant of tyrosine and DOPA Schiff bases were in the order of 104 to 108 M−1 suggesting a moderate to good binding ability with BSA. Higher binding affinity and better radical scavenging ability was observed for compounds bearing the naphthyl moiety.The radical scavenging ability of the l-DOPA naphthyl Schiff base was found to increase with decrease in IC50 values from 17 to 15 µM when bound to BSA. Fluorescence measurement and molecular docking studies demonstrated that hydrophobic interactions and hydrogen bonds dominated the interaction between BSA and tyrosine or l-DOPA derivatives. These results gave valuable information for pharmacological mechanism for l-tyrosine and l-DOPA Schiff bases in vivo conditions which can have a vital role in the development of drugs.