Cytotoxic Efficacy Research Articles

Design and synthesis of some new 6-bromo-2-(pyridin-3-yl)-4-substituted quinazolines as multi tyrosine kinase inhibitors

The present study involves design and synthesis of five series of 6-bromo-2-(pyridin-3-yl)-4-substituted quinazolines 9a-l, 11a-e, 13a-c, 14a-f and 15a-e. Candidates 9a-l and 11a-e were evaluated for their EGFR and HER2 inhibitory activity compared to Lapatinib. Compounds 9b, 9d, 9f, 11b and 11c were further screened for their in vitro cytotoxicity against two human breast cancer cell lines: AU-565 and MDA-MB-231 in addition to normal breast cell line MCF10A. Compound 9d revealed a remarkable cytotoxic efficacy against AU-565 cell line (IC50 = 1.54 µM) relative to Lapatinib (IC50 = 0.48 µM), whereas compounds 9d and 11c showed a superior cytotoxicity towards MDA-MB-231 (IC50 = 2.67 and 1.75 µM, respectively) in comparison to Lapatinib (IC50 = 9.29 µM). Moreover, compounds 13a-c, 13a-c, 14a-f and 15a-e were tested for their VEGFR-2 inhibitory activity compared to Sorafenib. Compounds 13a, 14c and 14e exhibited remarkable inhibition (IC50 = 79.80, 50.22 and 78.02 nM, respectively) relative to Sorafenib (IC50 = 51.87 nM). In vitro cytotoxicity of these compounds against HepG2, HCT-116 and normal cell (WISH) revealed a superior cytotoxicity against HepG2, HCT-116 especially 13a (IC50 = 17.51 and 5.56 µM, respectively) and 14c (IC50 = 10.40 and 3.37 µM, respectively) compared to Sorafenib (IC50 = 19.33 and 6.82 µM, respectively). Compounds 9d, 11c and 14c were subjected to cell cycle analysis and apoptotic assay. Molecular docking and ADME prediction studies were fulfilled to illustrate the interaction of the potent derivatives with the hot spots of the active site of EGFR, HER2 and VEGFR-2 along with prediction of their pharmacokinetic and physicochemical properties.

In Vitro Study of the Potential Role of Olive Oil Oleuropein in Modulating the 5-FU Cytotoxic Efficacy against the Tongue Squamous Cell Carcinoma

BACKGROUND: 5-fluorouracil (5-FU) is an anticancer drug used to inhibit the proliferation of many different tumor cells. Since severe side effects are associated with this drug, its combination with different natural compounds would allow the use of a significantly lower dose of 5-FU. Oleuropein (OLEU), has been shown to have inhibitory effects on various types of cancers. AIM: The main objective of the current study was to assess the cytotoxic effect of OLEU and the chemotherapeutic drug 5-FU on Human Tongue Carcinoma Cancer Cell Line (HNO-97) and Human Normal Oral Epithelial Cell Line (OEC) either independently or combinatory effect. MATERIALS AND METHODS: 3-(4,5- dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cell viability, and half-maximal inhibitory concentration (IC50) was calculated. Flowcytometry for cell cycle analysis was performed. Also, in vitro scratch assay was done to assess the inhibitory effects of OLEU on the migration of cells.RESULTS: MTT assay study demonstrated that OLEU and 5-FU alone or in combinations have produced a significant inhibitory effect on both normal and cancer cell lines with a favorable impact for OLEU on cancer cell lines rather than the normal one. A significant increase in the cell inhibitory % was reported between the single and the combinations treated groups as compared to the non-treated control group. Cell cycle analysis via flowcytometry showed that OLEU had induced cell cycle arrest at G0/1 phase, decreased S phase and G2/M phase either independently or in combination for 24h and 48h when compared with a non-treated control group. A Scratch assay test showed that OLEU could induce delayed wound healing. CONCLUSIONS: The findings of the present study suggest that OLEU can exert an anti-cancer effect on HNO-97 and may have the potential for potentiation of 5-FU cytotoxic effects and reduction of its adverse effects. In addition, OLEU could inhibit cancer progression and expansion from the initial tumor.

Validation of a Mathematical Model Describing the Dynamics of Chemotherapy for Chronic Lymphocytic Leukemia In Vivo.

In recent years, mathematical models have developed into an important tool for cancer research, combining quantitative analysis and natural processes. We have focused on Chronic Lymphocytic Leukemia (CLL), since it is one of the most common adult leukemias, which remains incurable. As the first step toward the mathematical prediction of in vivo drug efficacy, we first found that logistic growth best described the proliferation of fluorescently labeled murine A20 leukemic cells injected in immunocompetent Balb/c mice. Then, we tested the cytotoxic efficacy of Ibrutinib (Ibr) and Cytarabine (Cyt) in A20-bearing mice. The results afforded calculation of the killing rate of the A20 cells as a function of therapy. The experimental data were compared with the simulation model to validate the latter’s applicability. On the basis of these results, we developed a new ordinary differential equations (ODEs) model and provided its sensitivity and stability analysis. There was excellent accordance between numerical simulations of the model and results from in vivo experiments. We found that simulations of our model could predict that the combination of Cyt and Ibr would lead to approximately 95% killing of A20 cells. In its current format, the model can be used as a tool for mathematical prediction of in vivo drug efficacy, and could form the basis of software for prediction of personalized chemotherapy.

Chemical composition and cytotoxic activity of Callistemon citrinus (Myrtaceae) volatile oil and active fraction

Background: Volatile oils have found use locally in the management of many diseases including tumor-related ailments. Due to the short-comings of orthodox medicine, there is a need to source for alternative drugs with better effects. Callistemon citrinus oil contains some bioactive compounds which are useful in treating many diseases. This study was designed to examine the chemical composition and cytotoxic efficacy of this plant oil.Method: Callistemon citrinus volatile oil was extracted from fresh leaves using a Clavenger apparatus by hydro-distillation method. Preliminary cytotoxic screening was carried out with brine shrimp at 10-1000 μg/mL. The essential oil was further tested on breast (AU 565) and cervical (HeLa) cancer cell lines at 50 μg /mL using MTT assay. Column chromatography of the oil was carried out and the resulting fractions subjected to biological testing. GCMS analysis was carried out on the oil and the most active fraction.Results: The oil produced concentration- dependent activity with an LC50 of 528.48 μg/mL in the brine shrimp mortality assay. The oil also produced -7.60 and +11.80 % inhibitions against HeLa and AU 565 cells respectively. Column fraction F1 produced the highest activity against AU 565 cells with 70.44 % inhibition and an IC50 of 15.96 μg/mL. C. citrinus oil revealed the presence of cineole (36.06 %) and α-pinene (21.41 %) as the major components while 1,1'-(5-hydroxy-2,2-dimethylbicyclo[4.1.0]heptane-1,7-diyl)bis-, (1α,5β,6α,7α)-ethanone (24.89 %.) was the most abundant in the active fraction.Conclusion: C. citrinus volatile oil has cytotoxic potential and is a good candidate for further in vivo studies.

In Vitro Evaluation of Antioxidant, Anticancer, and Anti-Inflammatory Activities of Ethanolic Leaf Extract of Adenium obesum.

Adenium obesum commonly known as “desert rose” belongs to the family Apopcynaceae and has previously been reported for its anti-influenza, antimicrobial, and cytotoxic efficacies and well-known for their ethno-medicinal applications. In the present study, ethanolic extracts of A. obesum (AOE) were analyzed by gas chromatography-mass spectrometry (GC–MS) to identify the important phytochemical compounds. The GC–MS analysis of AOE detected the presence of 26 phytochemical compounds. This plant is traditionally used for the treatment of various diseases. In this report, the antioxidant, anti-inflammatory, and anticancer activities of ethanolic leaf extract from A. obesum (AOE) were studied. The antioxidant potential of ethanolic extract of AOE was examined by different antioxidant assays, such as antioxidant capacity by the DPPH, ABTS, superoxide, hydroxyl radical scavenging, and lipid peroxidation inhibition assays. The antioxidant activities of various reaction mixtures of AOE were compared with a reference or standard antioxidant (ascorbic acid). In addition, we also evaluated the anticancer activity of AOE, and it was observed that AOE was found to be cytotoxic against A549 lung cancer cells. It was found that AOE inhibited the viability of A549 lung cancer cells by inducing nuclear condensation and fragmentation. Furthermore, ethanolic AOE demonstrated the anti-inflammatory potential of AOE in murine alveolar macrophages (J774A.1) as an in vitro model system. AOE showed its potential in reducing the levels of inflammatory mediators including the proinflammatory cytokines and TNF-α. The results obtained in the present investigation established the antioxidant, anticancer, and anti-inflammatory potency of AOE, which may account for subsequent studies in the formulation of herbal-based medicine.

Triterpenoids from Stem Bark of <i>Dysoxylum excelsum</i> and Their Cytotoxic Activity against MCF-7 Breast Cancer Cells

The diversity of structures of triterpenoids provides a variety of interesting pharmacological effects, one of them being anticancer. Sources of triterpenoids are found in Dysoxylum excelsum, it is still rarely studied, and it potentially contains triterpenoids with cytotoxic activity. Therefore, the research aimed to isolate the triterpenoids from the stem bark of D. excelsum and determine their cytotoxic efficacy against the breast cancer cells of MCF-7. The dried stem bark of D. excelsum was macerated using methanol and then fractionated with n-hexane and ethyl acetate to obtain the corresponding fractions. Two triterpenoids, 3α,7α-dihydroxyapotirucalla-14,24-Z-dien-26-oic acid (1) and masticadienonic acid or 3-oxo-tirucalla-7,24-Z-dien-26-oic acid (2) were isolated in the first place from the Dysoxylum genus. Compounds 1 and 2 were assayed for their cytotoxic potency anti- MCF-7 using the MTT assay and gave moderate IC50 values of 60.87 and 72.86 µM, respectively. Based on their analogs, partial structure modification of the compounds might increase the activity; thus, it can be further utilized as better lead compounds for anticancer drugs candidate.

Interleukin Inhibitors in Cytokine Release Syndrome and Neurotoxicity Secondary to CAR-T Therapy.

Introduction: Chimeric antigen receptor T-cell (CAR-T) therapy is an innovative therapeutic option for addressing certain recurrent or refractory hematological malignancies. However, CAR-T cells also cause the release of pro-inflammatory cytokines that lead to life-threatening cytokine release syndrome and neurotoxicity. Objective: To study the efficacy of interleukin inhibitors in addressing cytokine release syndrome (CRS) and neurotoxicity secondary to CAR-T therapy. Methodology: The authors conducted a bibliographic review in which 10 articles were analyzed. These included cut-off studies, case reports, and clinical trials involving 11 cancer centers and up to 475 patients over 18 years of age. Results: Tocilizumab is the only interleukin inhibitor approved to address CRS secondary to CAR-T therapy due to its efficacy and safety. Other inhibitors, such as siltuximab and anakinra, could be useful in combination with tocilizumab for preventing severe cytokine release and neurotoxicity. In addition, the new specific inhibitors could be effective in mitigating CRS without affecting the cytotoxic efficacy of CAR-T therapy. Conclusion: More lines of research should be opened to elucidate the true implications of these drugs in treating the side effects of CAR-T therapy.

Miniaturized and multiplexed high-content screening of drug and immune sensitivity in a multichambered microwell chip

SummaryHere, we present a methodology based on multiplexed fluorescence screening of two- or three-dimensional cell cultures in a newly designed multichambered microwell chip, allowing direct assessment of drug or immune cell cytotoxic efficacy. We establish a framework for cell culture, formation of tumor spheroids, fluorescence labeling, and imaging of fixed or live cells at various magnifications directly in the chip together with data analysis and interpretation. The methodology is demonstrated by drug cytotoxicity screening using ovarian and non-small cell lung cancer cells and by cellular cytotoxicity screening targeting tumor spheroids of renal carcinoma and ovarian carcinoma with natural killer cells from healthy donors. The miniaturized format allowing long-term cell culture, efficient screening, and high-quality imaging of small sample volumes makes this methodology promising for individualized cytotoxicity tests for precision medicine.

Natural Killer Cell Recognition and Control of Epithelial Cancers.

Natural killer (NK) cells possess an innate ability to recognize cancer and are key mediators of cytotoxic efficacy for anticancer antibodies. Recent advances in the ability to generate, qualify, and safely infuse NK cells have led to a wide variety of clinical trials in oncology. Although their efficacy is best established for liquid cancers, their potential application in solid cancers has received increased attention. Here, we provide general background across a disparate group of exemplary solid tumors for which there is evidence for an NK cell role, discuss NK cell recognition motifs specific to each and murine and human studies of each that are supportive of NK cell adoptive immunotherapy, and end with special considerations relevant to the solid tumor microenvironment.

Role of Fc Core Fucosylation in the Effector Function of IgG1 Antibodies.

The presence of fucose on IgG1 Asn-297 N-linked glycan is the modification of the human IgG1 Fc structure with the most significant impact on FcɣRIII affinity. It also significantly enhances the efficacy of antibody dependent cellular cytotoxicity (ADCC) by natural killer (NK) cells in vitro, induced by IgG1 therapeutic monoclonal antibodies (mAbs). The effect of afucosylation on ADCC or antibody dependent phagocytosis (ADCP) mediated by macrophages or polymorphonuclear neutrophils (PMN) is less clear. Evidence for enhanced efficacy of afucosylated therapeutic mAbs in vivo has also been reported. This has led to the development of several therapeutic antibodies with low Fc core fucose to treat cancer and inflammatory diseases, seven of which have already been approved for clinical use. More recently, the regulation of IgG Fc core fucosylation has been shown to take place naturally during the B-cell immune response: A decrease in α-1,6 fucose has been observed in polyclonal, antigen-specific IgG1 antibodies which are generated during alloimmunization of pregnant women by fetal erythrocyte or platelet antigens and following infection by some enveloped viruses and parasites. Low IgG1 Fc core fucose on antigen-specific polyclonal IgG1 has been linked to disease severity in several cases, such as SARS-CoV 2 and Dengue virus infection and during alloimmunization, highlighting the in vivo significance of this phenomenon. This review aims to summarize the current knowledge about human IgG1 Fc core fucosylation and its regulation and function in vivo, in the context of both therapeutic antibodies and the natural immune response. The parallels in these two areas are informative about the mechanisms and in vivo effects of Fc core fucosylation, and may allow to further exploit the desired properties of this modification in different clinical contexts.

Anti-EpCAM Functionalized I-131 Radiolabeled Biomimetic Nanocarrier Sodium/Iodide-Symporter-Mediated Breast-Cancer Treatment.

Currently, breast-cancer treatment has a number of adverse side effects and is associated with poor rates of progression-free survival. Therefore, a radiolabeled anti-EpCAM targeted biomimetic coated nanocarrier (EINP) was developed in this study to overcome some of the treatment challenges. The double emulsion method synthesized the poly(lactic-co-glycolic acid) (PLGA) nanoparticle with Na131I entrapped in the core. The PLGA nanoparticle was coated in human red blood cell membranes and labeled with epithelial cell adhesion molecule (EpCAM) antibody to enable it to target EpCAM overexpression by breast-cancer cells. Characterization determined the EINP size as 295 nm, zeta potential as −35.9 mV, and polydispersity as 0.297. EINP radiochemical purity was >95%. Results determined the EINP efficacy against EpCAM positive MCF-7 breast cancer at 24, 48, and 72 h were 69.11%, 77.84%, and 74.6%, respectively, demonstrating that the EINPs achieved greater cytotoxic efficacy supported by NIS-mediated Na131I uptake than the non-targeted 131INPs and Na131I. In comparison, fibroblast (EpCAM negative) treated with EINPs had significantly lower cytotoxicity than Na131I and 131INPs (p < 0.05). Flow cytometry fluorescence imaging visually signified delivery by EINPs specifically to breast-cancer cells as a result of anti-EpCAM targeting. Additionally, the EINP had a favorable safety profile, as determined by hemolysis.

Preparation of carrageenan/ chitosan-based (N,N,N-trimeth(yl chitosan chloride) silver nanocomposites as pH sensitive carrier for effective controlled curcumin delivery in cancer cells

PurposeThis work aims to formulate a novel pH-sensitive biocompatible metallic nanocomposite as a carrier for hydrophobic drugs with efficient loading and controlled drug release. The proposed carrier consists of a polyelectrolyte complex of chitosan-based, N,N,N-trimethyl chitosan chloride (TMC), and carrageenan (CAR) biopolymers with different silver nanoparticles (AgNPs) molar ratios forming CAR/TMC-Ag nanocomposites that investigated to be used as a carrier for controlled gastro-intestinal tract (GIT) release of curcumin (CUR) as a model of lipophilic drugs. MethodsThe structure of nanocomposites was scrutinized via FTIR, TEM, SEM, XRD and EDX, whereas the potential cytotoxicity and antimicrobial efficacy of loaded nanocomposite with 3% AgNPs (NC-3% Ag) were probed, as well. ResultsSuccessful loading and release of CUR were detected from the prepared nanocomposites without change in its functionality with a higher releasing profile in pH 7.4 (simulated intestinal fluid) than pH 1.2 (simulated gastric fluid). In vitro release study of CUR loaded NC-3% Ag showed the favored sustained drug release reaching 98.9% ± 0.9 within 24 h in pH 7.4, following Korsmeyer−Peppas model. Additionally, successful release from loaded CAR/TMC nanocomposite with 3% AgNPs (NC-3% Ag), revealed high cytotoxic effect with apoptotic induction against human colon cancerous cells (Caco-2) through G2/M cell cycle arrest and antibacterial potency against different bacterial strains. Moreover, cell viability results of NC-3% Ag against human normal cells exhibited good biocompatibility. ConclusionAccordingly, we suggested NC-3% Ag as a promising controlled drug delivery vehicle for hydrophobic drugs with preferred intestinal targeting effectiveness.

An efficient p‐TSA catalyzed synthesis of some new substituted‐(5‐hydroxy‐3‐phenylisoxazol‐4‐yl)‐1,3‐dimethyl‐1H‐chromeno[2,3‐d]pyrimidine‐2,4(3H,5H)‐dione/3,3‐dimethyl‐2H‐xanthen‐1(9H)‐one scaffolds and evaluation of their pharmacological and computational investigations

We have developed an easy and efficient protocol for the synthesis of a series of some novel substituted-(5-hydroxy-3-phenylisoxazol-4-yl)-1,3-dimethyl-1H-chromeno[2,3-d]pyrimidine-2,4(3H,5H)-dione/3,3-dimethyl-2H-xanthen-1(9H)-one derivatives (4a–j) via p-TSA catalyzed reaction of 1,3-dimethylbarbituric acid/dimedone, substituted salicylaldehyde/2-hydroxy-nepthaldehyde and 3-phenyl-5-isoxazolone was administered in refluxed temperature in the presence of aqueous ethanol. All the obtained compounds were evaluated for their therapeutic effect using pharmacological and computational investigations, and structures were confirmed using analytical and spectroscopic techniques. Absorption spectra were recorded in six different solvents such as polar protic, polar aprotic, and nonpolar solvents, λmax of all the compounds are appeared at bathochromic shift towards the longer wavelength due to the π-π* & n-π* transitions. The results of pharmacological investigations revealed that the compounds 4c, 4e, and 4h possess excellent cytotoxicity efficacy, and compounds 4c, 4d, 4h, and 4i have shown better anti-TB efficiency. The SAR study shows the importance of the electron-withdrawing group and additional phenyl nucleus enhancing the biological potency of the compounds. The results of the in silico molecular docking studies revealed that the compounds 4c and 4h effectively interacted with P38 MAP kinase (-10.9 kcal/mol) and InhA-Enoyl-Acyl Carrier Protein Reductase (-11.0 kcal/mol), respectively. Further, the molecular dynamics (MD) simulation studies revealed that the compounds 4c and 4h stabilized the macromolecular structures in terms of RMSD, RMSF, the Radius of gyration, and SASA compared to their unbound and standard compound bound structures. DFT study suggested that the compounds are chemically and biologically more reactive due to less energy gap.

Abstract 5473: Isoform-selective HDAC inhibition up-regulates CD26 expression on multiple myeloma cells and augments cytotoxic efficacy by humanized monoclonal antibody

Abstract Objective: CD26, a 110-kDa transmembrane glycoprotein, which is expressed on several tumor cells including malignant lymphoma, has been implicated in tumorigenesis, whereas, its roles in plasma cell malignancies remain elusive. Recently, we have identified that CD26 is uniformly and intensely expressed in osteoclasts, while its expression in plasma cells of patients with multiple myeloma (MM) reveals heterogenous. Decreased expression levels of CD26 in MM cells are one of the mechanisms underlying resistance to humanized anti-CD26 monoclonal antibody (mAb) therapy. In the present study, we clarify the impact of epigenetic modification by HDAC inhibition (HDACi) with isoform-selective or broad inhibitors on the regulation of CD26 in MM cells and its mechanisms, thereby affecting the performance of humanized CD26mAb. Methods and Results: Immunostaining of bone marrow tissues of MM showed that CD26pos/CD138pos plasma cells were detected in several patients but not in others. So, first, we investigated the impact of HDACi on CD26 expression of MM cells. Although the cell surface expression of CD26 was relatively low or not detected on 5 MM cell lines (KMS26, KMS27, KMS28, KMS11, RPMI8226), the increased expression in CD26 levels was detectable within 24 h of the treatment with HDAC1i; FK228, HDAC3i; BG45, MS-275, RG2833 or HDAC6i; nexturastat A, tubastatin A, ACY-1215 as well as broad HDACi; LBH-589, SAHA. It increased further and maximum increase was observed at 72 h of the treatment by each HDACi. Then, subsequent removal of HDACi resulted in a decline of CD26 expression on MM cells to pre-treated levels. In addition, the levels of CD26mRNA were concomitantly enhanced, which was paralleled with an increase in the induction of CD26 protein in MM cells. Next, we examined the effect of HDACi on the viability of CD26neg MM cells in the absence or presence of CD26mAb. The mAb alone did not induce lysis of CD26neg MM cell lines at any doses, whereas, combining HDACi plus CD26mAb (10 μg/ml) synergistically facilitated lysis of CD26neg MM cells via direct effects as well as NK cell-mediated ADCC by mAb. Of note, HDACi plus CD26mAb in combination readily augmented lysis of CD26neg cell populations, refractory to HDACi or mAb alone. Furthermore, to elucidate the mechanisms of CD26 up-regulation in MM cells by HDACi, we performed chromatin immunoprecipitation assay on CD26 gene promoter. Each HDACi increased acetylation of histone 3 lysine 27 (H3K27Ac), concomitant with enhanced binding of Sp1 at the 5’ flanking region of the CD26 gene containing Sp1 binding sites in CD26neg MM cells, which is suggestive of transactivation of CD26 gene by HDACi targeting HDAC1, 3 or 6. Conclusion: Combination with isoform-selective HDACi not only shows anti-MM activity but supports as immunopotentiators by sensitizing CD26neg MM cells to CD26mAb and augment its cytotoxicity against MM cells. Citation Format: Hiroko Nishida, Mari Fujiwara, Mutsumi Hayashi, Michiie Sakamoto. Isoform-selective HDAC inhibition up-regulates CD26 expression on multiple myeloma cells and augments cytotoxic efficacy by humanized monoclonal antibody [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5473.

Factors Influencing the Therapeutic Efficacy of the PSMA Targeting Radioligand 212Pb-NG001.

Simple SummaryProstate-specific membrane antigen (PSMA) is a protein overexpressed in metastatic castration-resistant prostate cancer and a promising target for targeted radionuclide therapy. PSMA-targeted alpha therapy is of growing interest due to the high-emission energy and short range of alpha particles, resulting in a prominent cytotoxic potency. This study assesses the influence of various factors on the in vitro and in vivo therapeutic efficacy of the alpha particle generating PSMA-targeting radioligand 212Pb-NG001.This study aimed to determine the influence of cellular PSMA expression, radioligand binding and internalization, and repeated administrations on the therapeutic effects of the PSMA-targeting radioligand 212Pb-NG001. Cellular binding and internalization, cytotoxicity, biodistribution, and the therapeutic efficacy of 212Pb-NG001 were investigated in two human prostate cancer cell lines with different PSMA levels: C4-2 (PSMA+) and PC-3 PIP (PSMA+++). Despite 10-fold higher PSMA expression on PC-3 PIP cells, cytotoxicity and therapeutic efficacy of the radioligand was only 1.8-fold better than for the C4-2 model, possibly explained by lower cellular internalization and less blood-rich stroma in PC-3 PIP xenografts. Mice bearing subcutaneous PC-3 PIP xenografts were treated with 0.2, 0.4, and 0.8 MBq of 212Pb-NG001 that resulted in therapeutic indexes of 2.7, 3.0, and 3.5, respectively. A significant increase in treatment response was observed in mice that received repeated injections compared to the corresponding single dose (therapeutic indexes of 3.6 for 2 × 0.2 MBq and 4.4 for 2 × 0.4 MBq). The results indicate that 212Pb-NG001 can induce therapeutic effects at clinically transferrable doses, both in the C4-2 model that resembles solid tumors and micrometastases with natural PSMA expression and in the PC-3 PIP model that mimics poorly vascularized metastases.

Anticancer activity of bioactive compound chavicol as potential toxic against human lung cancer A549 cells

Cellular breakdown in the lungs is the most well-known human disease, and it is also the leading cause of death. The majority of anticancer therapies have substantial toxicity and serious health issues because of the multidrug resistance upon treatment with various drugs. Thus development of successful therapeutic solutions remains as a key factor in the pharmaceuticals. For the effective treatment of cancer, bioactive substances are the most essential in the domain of therapeutic use as they have minimal or no side effects to the host. In this study, we explore the in vitro anti-cancer efficacy of bioactive compound chavicol against human lung cancer cells (A-549). The treatment with chavicol compound inhibited cancer cell growth, survival, migration, and invasion. In A-549 cells, the cytotoxic efficacy of chavicol was shown to be significant with about 31 μg/mL while the normal cells had not shown any significant adverse effect. The morphological alterations related with apoptosis were observed using acridine orange and ethidium bromide staining (AO/EtBr) with various doses of chavicol. Following that, using fluorescence microscope analysis, the effect of chavicol on cell cycle stage analysis and mitochondrial membrane potential was studied. According to the current findings, chavicol therapy led to loss of mitochondrial membrane potential stability in A-549 cells. Overall, this study had clearly shown the potential exploration of bioactive compound chavicol in the therapeutic use against lung cancer.

Construction, pH-responsive drug release and in vitro cytotoxicity of Ni@C nanotubes

Ni@C magnetic nanotubes surrounded by a carbon shell with thick of 10 nm were constructed using nickel nanotubes as core by glucose-solvothermal-method. The morphology, structure, composition, magnetic property, drug release behavior, in-vitro cytotoxicity and antitumor efficacy of the as-prepared Ni@C nanotubes were studied by scanning electron microscopy, transmission electron microscopy, X-ray diffraction, Raman spectrum, energy dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, vibrating-sample magnetometer, UV–vis spectroscopy, CCK-8 assays and flow cytometry. The results indicated that the as-prepared Ni@C nanotubes had typical hollow core-shell structure, good magnetic property, pH-responsive drug-release functionality and better antitumor activity, compared with nickel nanotubes. It presented low in-vitro cytotoxicity originated in cell apoptosis by S-phase arrest, which made the magnetic Ni@C nanotubes potentially application in biomedicine.

Application of blood brain barrier models in pre-clinical assessment of glioblastoma-targeting CAR-T based immunotherapies

Human blood brain barrier (BBB) models derived from induced pluripotent stem cells (iPSCs) have become an important tool for the discovery and preclinical evaluation of central nervous system (CNS) targeting cell and gene-based therapies. Chimeric antigen receptor (CAR)-T cell therapy is a revolutionary form of gene-modified cell-based immunotherapy with potential for targeting solid tumors, such as glioblastomas. Crossing the BBB is an important step in the systemic application of CAR-T therapy for the treatment of glioblastomas and other CNS malignancies. In addition, even CAR-T therapies targeting non-CNS antigens, such as the well-known CD19-CAR-T therapies, are known to trigger CNS side-effects including brain swelling due to BBB disruption. In this study, we used iPSC-derived brain endothelial-like cell (iBEC) transwell co-culture model to assess BBB extravasation of CAR-T based immunotherapies targeting U87MG human glioblastoma (GBM) cells overexpressing the tumor-specific mutated protein EGFRvIII (U87vIII). Two types of anti-EGFRvIII targeting CAR-T cells, with varying tonic signaling profiles (CAR-F263 and CAR-F269), and control Mock T cells were applied on the luminal side of BBB model in vitro. CAR-F263 and CAR-F269 T cells triggered a decrease in transendothelial electrical resistance (TEER) and an increase in BBB permeability. CAR-T cell extravasation and U87vIII cytotoxicity were assessed from the abluminal compartment using flow cytometry and Incucyte real-time viability imaging, respectively. A significant decrease in U87vIII cell viability was observed over 48 h, with the most robust cytotoxicity response observed for the constitutively activated CAR-F263. CAR-F269 T cells showed a similar cytotoxic profile but were approximately four fold less efficient at killing the U87vIII cells compared to CAR-F263, despite similar transmigration rates. Visualization of CAR-T cell extravasation across the BBB was further confirmed using BBTB-on-CHIP models. The described BBB assay was able to discriminate the cytotoxic efficacies of different EGFRvIII-CARs and provide a measure of potential alterations to BBB integrity. Collectively, we illustrate how BBB models in vitro can be a valuable tool in deciphering the mechanisms of CAR-T–induced BBB disruption, accompanying toxicity and effector function on post-barrier target cells.

In-silico predicting as a tool to develop plant-based biomedicines and nanoparticles: Lycium shawii metabolites

In silico approach helps develop biomedicines and is useful for exploring the pharmacology of potential therapeutics using computer-simulated models. In vitro assays were used to determine the anti-microbial and cytotoxic efficacies of silver nanoparticles (AgNPs) synthesized with the shrub Lycium shawii. In silico predicting was performed to assess the L. shawii metabolites identified using QTOF-LCMS for their pharmacological properties. L. shawii mediated AgNPs were synthesized and characterized (FTIR, TEM, SEM, DLS and EDX). The anti-bacterial efficacies of L. shawii extract, AgNPs, and penicillin-conjugated AgNPs (pen-AgNPs) were determined. The cytotoxicity of the AgNPs was measured against colorectal cancer cell line (HCT116), normal breast epithelium (MCF 10A), and breast cancer cell line (MDA MB 231). Five molecules (costunolide, catechin, emodin, lyciumaside, and aloe emodin 11-O-rhamnoside) were detected in the L. shawii extract. AgNPs (69nm) were spherical with crystallographic structure. All three agents prepared showed inhibitory activity against the tested bacteria, the most efficacious being pen-AgNPs. High cytotoxicity of AgNPs (IC50 62μg/ml) was observed against HCT116, IC50 was 78μg/ml for MCF 10A, and 250μg/ml for MDA MB 231, of which cells showed apoptotic features under TEM examination. The in silico approach indicated that the carbonic anhydrase IX enzyme was the target molecule mediating anti-cancer and anti-bacterial activities and that emodin was the metabolite in action. Combining in vitro studies and in silico molecular target prediction helps find novel therapeutic agents. Among L. shawii metabolites, emodin is suggested for further studies as an agent for drug development against pathogenic bacteria and cancer.

Enhanced in vitro cytotoxicity and antitumor activity in vivo of iridium(III) complexes liposomes targeting endoplasmic reticulum and mitochondria

In this paper, two new iridium(III) complexes [Ir(ppy)2(CBIP)](PF6) (ppy = 2-phenylpyridine, CBIP = 2-(4′-chloro-(1,1′-biphenyl))-1H-imidazo[4,5-f][1,10]phenanthroline) (Ir1) and [Ir(piq)2(CBIP)](PF6) (piq = 1-phenylisoquinoline) (Ir2) were synthesized and characterized. The anticancer activity of the complexes against cancer A549, HepG2, SGC-7901, BEL-7402, HeLa and LO2 cells was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Unexpectedly, the complexes exhibit no or low cytotoxic activity toward the selected cancer cells. To increase the anticancer activity, complexes Ir1 and Ir2 were encapsulated into the liposome to form Ir1lipo and Ir2lipo, while Ir1lipo and Ir2lipo show high cytotoxic efficacy against BEL-7402, SGC-7901 and HeLa cells and Ir2lipo displays moderate cytotoxic activity against A549 and HepG2. The anticancer mechanism was explored through wound healing, cell cycle arrest, apoptosis, the change of mitochondrial membrane potential and antitumor activity in vivo. The antitumor in vivo showed that Ir1Lipo (3.9 mg/kg) exhibited significant antitumor activity with an inhibitory rate of 62.16%. Additionally, the expression of B-cell lymphoma-2 family proteins was studies by western blotting analysis. The results demonstrate that Ir1lipo and Ir2lipo induce apoptosis in BEL-7402 cells via endoplasmic reticulum stress-mitochondrial pathway.