Survival Time Of Animals Research Articles

Emodin Enhances the Chemosensitivity of Endometrial Cancer by Inhibiting ROS-Mediated Cisplatin-resistance.

Endometrial cancer is a common cause of death in gynecological malignancies. Cisplatin is a clinically chemotherapeutic agent. However, drug-resistance is the primary cause of treatment failure. Emodin is commonly used clinically to increase the sensitivity of chemotherapeutic agents, yet whether Emodin promotes the role of Cisplatin in the treatment of endometrial cancer has not been studied. CCK-8 kit was utilized to determine the growth of two endometrial cancer cell lines, Ishikawa and HEC-IB. The apoptosis level of Ishikawa and HEC-IB cells was detected by Annexin V / propidium iodide double-staining assay. ROS level was detected by DCFH-DA and NADPH oxidase expression. Expressions of drug-resistant genes were examined by real-time PCR and Western blotting. Emodin combined with Cisplatin reduced cell growth and increased the apoptosis of endometrial cancer cells. Co-treatment of Emodin and Cisplatin increased chemosensitivity by inhibiting the expression of drugresistant genes through reducing the ROS levels in endometrial cancer cells. In an endometrial cancer xenograft murine model, the tumor size was reduced and animal survival time was increased by co-treatment of Emodin and Cisplatin. This study demonstrates that Emodin enhances the chemosensitivity of Cisplatin on endometrial cancer by inhibiting ROS-mediated expression of drug-resistance genes.

Systematic review and meta-analysis of the protective effect of resveratrol on multiple organ injury induced by sepsis in animal models.

Sepsis may directly lead to multiple organ failure, which is among the leading causes of mortality in critically ill patients. According to data released by the Global Sepsis Alliance, the number of mortalities due to sepsis exceeded the combined number for prostate cancer, breast cancer and AIDS in 2012. To date, studies have reported that resveratrol has marked positive effects including anti-inflammatory, anti-oxidative and pro-microcirculatory functions in sepsis-induced organ injury, significantly improving the survival time and mortality of sepsis animals. The present systematic review sought to further clarify the efficacy and safety of resveratrol in the treatment of sepsis. Studies on resveratrol application in the treatment of sepsis-induced organ injury in animal models were reviewed by searching various Chinese and other language databases (PubMed, Embase, CNKI, WanFang and WeiPu) and by manually searching the references of related articles. The selection and evaluation of the studies was performed by two independent reviewers. A total of 260 related studies were initially identified. Following application of the exclusion factors and inclusion criteria, 11 studies were included. Meta-analysis revealed that resveratrol exerted significant protective effect in sepsis-induced animal models of organ injury, through anti-inflammatory, anti-oxidant and pro-microcirculatory functions compared with in the placebo group. While nuclear factor κB (NF-κB) and nuclear factor E2-related factor 2 (NRF-2) are the two major signaling pathways to have been associated with the anti-inflammatory and anti-oxidative effects of resveratrol, these factors were not quantified for mean values, therefore not suitable for systematic evaluation. For related factors, the results of meta-analysis were as follows: For tumor necrosis factor-α (TNF-α), the standardized mean difference (SMD) was -13.50 [95% confidence interval (CI): -22.08, -4.91; P=0.002]; for malondialdehyde (MDA), the SMD was -3.10 (95% CI: -5.27, -0.93; P=0.005); for mean arterial pressure the SMD was 1.34 (95% CI: 0.07, 2.62; P=0.04); for interleukin (IL)-6 the SMD was -9.57 (95% CI: -20.90, 1.75; P=0.10); and for IL-10 the SMD was 0.80 (95% CI: -0.73, 2.34; P=0.31). It was concluded that resveratrol exerted significant anti-inflammatory and anti-oxidative effects through NF-κB and NRF-2 signaling pathways in animal models of sepsis-induced multiple organ injury, manifesting as significant downregulation of TNF-α and MDA expression and improved microcirculation, therefore ameliorating septic damage to the body, which may ultimately improve survival ratios.

Curative effect of radiofrequency ablation for firearm injury in livers combined with seawater immersion

Objective To observe the curative effect of radiofrequency ablation for fireman injury in livers combined with seawater immersion. Methods Ten adult male beagles were selected as the research objects. The method of surgical operation exposure combined with shooting by QSZ-92 training pistol was used to cause fireman injury in the left lobe of liver, and then animals were soaked in the seawater for 5 min. Radiofrequency ablation was applied to control hemorrhage of injured liver following animal was salvaged from seawater. The survival time of animals was recorded and the survival rate was statistically analyzed; the rectal temperature was monitored and compared before and after the seawater immersion. Measurements of complete blood count and blood chemistries were performed at baseline, 3rd, 7th and 14th postoperative day. Results All animals survived to 14th postoperative day, with a survival rate of 100%. The rectal temperature decreased significantly after seawater immersion (P=0.000). There was no significant difference in erythrocyte count, hemoglobin concentration and platelet count at baseline, 3rd, 7th and 14th postoperative day (P=0.081, 0.264, 0.070). The level of alanine aminotransferase (ALT) at 3rd postoperative day was significantly higher than baseline (P=0.000); the level of ALT at 7th postoperative day was significantly lower than that at 3rd postoperative day (P=0.000), but still higher than baseline (P=0.000); the level of ALT restored to normal till 14th postoperative day (P=0.218). The level of aspartate aminotransferase (AST) at 3rd postoperative day was significantly higher than baseline (P=0.002), and the level of AST restored to normal at 7th postoperative day (P=0.290). The concentration of serum albumin at 3rd postoperative day was significantly lower than baseline (P=0.005), and didn’t restore to normal at 14th postoperative day (P=0.010). Conclusion In this study, a large-scale animal model of firearm injury in liver combined with seawater immersion was successfully established, and the hemostasis efficacy of radiofrequency ablation on such trauma was confirmed. Key words: Liver; Naval warfare injury; Radiofrequency ablation; Curative effect

Highlights of This Issue

Sequential treatment with targeted therapies can result in complex combinations of resistance mutations in drug targets. To establish the origins of resistance mutations, Schmitt and colleagues integrated single-molecule duplex sequencing of Philadelphia-positive leukemias with computational simulations. A combination of drug target mutation burden and tumor-initiating cell fraction predicts the likelihood of preexisting resistance mutations. Compound resistance is unlikely to exist before therapy, but rather is predicted to arise following the failure of first-generation therapies. The results predict that front-line use of pan-target therapies (i.e., therapies that can overcome any single resistance mutation) could improve outcomes.Correlation of HER2 expression to therapeutic resistance was previously investigated on ex vivo models by others. To explore the role of dynamic status of HER2 on tumor cells, Li and colleagues performed HER2-iFISH to longitudinally examine constant variation of HER2 phenotype and chromosome aneuploidy in CTCs in gastric cancer patients. Comparing to low positivity of histopathologic detection of HER2 in tumor mass, significantly higher positivity (>70%) of HER2 expression on aneuploid CTCs was demonstrated, which was relevant to developing resistance to the targeted and/or chemotherapy and thus shows promise to monitor developing therapeutic resistance in real time.Anti-PD-1/PD-L1-based immunotherapies can be transformative in a subset of metastatic cancer patients, but reliable biomarkers of response have been elusive. To integrate biomarkers of efficacy in clinical practice, Johnson and colleagues performed quantitative, multiplexed immunofluorescence tests comprised of HLA-DR, IDO-1, PD-1, and PD-L1 utilizing novel AQUA algorithms on archival, pre-treatment samples from 166 metastatic melanoma patients treated with PD-1 blockers across ten cancer centers. Biomarker signature-positive patients experienced superior survival and response rates in both discovery and validation cohorts. This two-slide assay on archival tissue may help identify patients who will have excellent outcomes with anti-PD-1 therapy.Pancreatic cancer is one of the biggest clinical challenges. Zhou and colleagues used a combination of local photothermal therapy and intratumoral application of an immunoadjuvant to treat a metastatic pancreatic tumor in mice, both subcutaneously and orthotopically. This unique combination significantly prolonged the animal survival time and slowed and, in many cases, prohibited the distant metastases. It also induced a long-term, tumor-specific immunity by increasing the quantity and quality of splenic and tumor-infiltrating antitumor T cells. This novel approach with its successful treatment results, particularly on orthotopic pancreatic tumors, shows promise as a clinical treatment modality for pancreatic cancer patients.

PO-177 Effect of rational exercise on delaying brain aging and its mechanism analysis

PO-177 Effect of rational exercise on delaying brain aging and its mechanism analysis

Design of multifunctional peptide collaborated and docetaxel loaded lipid nanoparticles for antiglioma therapy

Glioblastoma multiforme (GBM) is one of the most encountered gliomas of the central nervous system. The chemotherapeutic drugs used in the treatment of GBM suffer from poor blood brain barrier penetration, severe systemic toxicities and lack of specificity towards tumor cells. There is an urgent need to explore novel drug delivery systems specifically designed for targeting GBM. Solid lipid nanoparticles (SLN) are biocompatible vehicle with less toxicity issues compared to other drug delivery systems and serve the purpose of obviating the limitations posed by existing anti-cancer drugs for GBM. In this study, angiopep-2, a ligand for the lipoprotein receptor related protein 1 (LRP 1) receptor over expressed in endothelial cells of both brain and glioma, was grafted on the surface of solid lipid nanoparticles for the delivery of docetaxel. The peptide grafted nanoparticles (A-SLN) showed increased cytotoxicity, enhanced cellular internalization and prominent apoptosis than that of unconjugated nanoparticles against U87MG human glioblastoma and GL261 mouse glioma cells. A significant dual targeting effect of A-SLN (p < 0.0001) was confirmed in in-vivo studies by real time fluorescence imaging studies in glioblastoma induced C57BL/6 mice model. Pharmacokinetic and tissue distribution studies showed selective targeting with higher accumulation of A-SLN in brain compared to Taxtotere, a marketed formulation of docetaxel. After treatment with A-SLN, the mean animal survival time of the animals was significantly enhanced to 39 days from 24 days of plain docetaxel. Collectively, this study indicated that solid lipid nanoparticles decorated with angiopep-2 can be an excellent option as targeted drug delivery system for antiglioma therapy.

Development of a magnetic nano-graphene oxide carrier for improved glioma-targeted drug delivery and imaging: In vitro and in vivo evaluations

To overcome the obstacles inflicted by the BBB in Glioblastoma multiforme (GBM) we investigated the use of Multifunctional nanoparticles that designed with a Nano-graphene oxide (NGO) sheet functionalized with magnetic poly (lactic-co-glycolic acid) (PLGA) and was used for glioma targeting delivery of radiosensitizing 5-iodo-2-deoxyuridine (IUdR). In vitro biocompatibility of nanocomposite has been studied by the MTT assay. In vivo efficacy of magnetic targeting on the amount and selectivity of magnetic nanoparticles accumulation in glioma-bearing rats under an external magnetic field (EMF) density of 0.5 T was easily monitored with MRI. IUdR-loaded magnetic NGO/PLGA with a diameter of 71.8 nm, a zeta potential of −33.07 ± 0.07 mV, and a drug loading content of 3.04 ± 0.46% presented superior superparamagnetic properties with a saturation magnetization (Ms) of 15.98 emu/g. Furthermore, Prussian blue staining showed effective magnetic targeting, leading to remarkably improved tumor inhibitory efficiency of IUdR. The tumor volume of rats after treatment with IUdR/NGO/SPION/PLGA + MF was decreased significantly compared to the rats treated with buffer saline, IUdR and SPION/IUdR/NGO/PLGA. Most importantly, our data demonstrate that IUdR/NGO/SPION/PLGA at the present magnetic field prolongs the median survival time of animals bearing gliomas (38 days, p < 0.01). Nanoparticles also had high thermal sensitivities under the alternating magnetic field. In conclusion, we developed magnetic IUdR/NGO/PLGA, which not only achieved to high accumulation at the targeted tumor site by magnetic targeting but also indicated significantly enhanced therapeutic efficiency and toxicity for glioma both in vitro and in vivo. This innovation increases the possibility of improving clinical efficiency of IUdR as a radiosensitizer, or lowering the total drug dose to decrease systemic toxicity.

Prophylactic efficacy of some chemoprotectants against abrin induced lethality.

Abrin is a highly toxic protein produced by Abrus precatorius. Exposure to abrin, either through accident or by act of terrorism, poses a significant risk to human health and safety. Abrin functions as a ribosome-inactivating protein by depurinating the 28S rRNA and inhibits protein synthesis. It is a potent toxin warfare agent. There are no antidotes available for abrin intoxication. Supportive care is the only option for treatment of abrin exposure. It is becoming increasingly important to develop countermeasures for abrin by developing pre- and post-exposure therapy. The aim of this study is to screen certain pharmaceutical compounds for their chemoprotective properties against abrin toxicity in vivo in BALB/c male mice. Twenty-one compounds having either antioxidant, anti-inflammatory and cyto-protective properties or combination of them, were screened and administered as 1h pre-treatment followed by exposure of lethal dose (2×LD50, intraperitoneally) of abrin. To assess the protective efficacy of the compounds, survival and body weight was monitored. Fifteen compounds extended the survival time of animals significantly, as compared to abrin. The following five of these compounds, namely: Epicatechin-3-gallate, Gallic Acid, Lipoic Acid, GSH and Indomethacin extended the life time ranging from 6 to 9 days. These compounds also attenuated the abrin induced inflammation and enzymes associated with liver function, but none of them could prevent abrin induced lethality. The compounds offering extension of life could be useful to provide a time-window for other supportive treatment and could also be used as combinatorial therapy with other medical countermeasures against abrin induced lethality.

Antihypoxic activities of Crataegus pentaegyn and Crataegus microphylla fruits-an in vivo assay

The aim of this study was to evaluate anti-hypoxia activity of polyphenolic extracts of Crataegus microphylla and Crataegus pentaegyn on mice. Three experimental models of hypoxia were considered, including asphyctic hypoxia, haemic hypoxia, and circulatory hypoxia. Polyphenolic extract of both plants exhibited significant anti-hypoxic activity and prolonged animal survival time. Anti-hypoia activity of C. pentaegyn was found to be superior to that of C. microphylla in circulatory and asphyctic hypoxia. Antihypoxic activity of these extracts may be attributed to their phenolic compounds.

Metal nanoparticles restrict the growth of protozoan parasites

The Trypanosoma and Toxoplasma spp, are etiological agents of diseases capable of causing significant morbidity, mortality and economic burden, predominantly in developing countries. Currently, there are no effective vaccines for the diseases caused by these parasites; therefore, therapy relies heavily on antiprotozoal drugs. However, the treatment options for these parasitic diseases are limited, thus underscoring the need for new anti-protozoal agents. Here, we investigated the anti-parasite action of nanoparticles. We found that the nanoparticles have strong and selective in vitro activity against T. b. brucei but moderate in vitro activity against T. congolense and T. evansi. An estimation of the in vitro anti-Trypanosoma efficacy showed that the nanoparticles had ≥200-fold selective activity against the parasite versus mammalian cells. Moreover, the nanoparticle alloys moderately suppressed the in vitro growth of T. gondii by ≥60%. In our in vivo study, the nanoparticles appeared to exhibit a trypanostatic effect, but did not totally suppress the rat parasite burden, thereby failing to appreciably extend the survival time of infected animals compared with the untreated control. In conclusion, this is the first study to demonstrate the selective in vitro anti-Trypanosoma action of nanoparticles and thus supports the potential of nanoparticles as alternative anti-parasitic agents.

Differential In Vitro Cultivation of Francisella tularensis Influences Live Vaccine Protective Efficacy by Altering the Immune Response.

Francisella tularensis (Ft) is a biothreat agent for which there is no FDA-approved human vaccine. Currently, there are substantial efforts underway to develop both vaccines and improved tools to assess these vaccines. Ft expresses distinct sets of antigens (Ags) in vivo as compared to those expressed in vitro. Importantly, Ft grown in brain-heart infusion medium (BHIM) more closely mimics the antigenic profile of macrophage-grown Ft when compared to Mueller-Hinton medium (MHM)-grown Ft. Thus, we predicted that when used as a live vaccine BHIM-grown Ft (BHIM-Ft) would provide better protection, as compared to MHM-Ft. We first determined if there was a difference in growth kinetics between BHIM and MHM-Ft. We found that BHIM-Ft exhibited an initial growth advantage ex vivo that manifests as slightly hastened intracellular replication as compared to MHM-Ft. We also observed that BHIM-Ft exhibited an initial growth advantage in vivo represented by rapid bacterial expansion and systemic dissemination associated with a slightly shorter mean survival time of naive animals. Next, using two distinct strains of Ft LVS (WT and sodB), we observed that mice vaccinated with live BHIM-Ft LVS exhibited significantly better protection against Ft SchuS4 respiratory challenge compared to MHM-Ft-immunized mice. This enhanced protection correlated with lower bacterial burden, reduced tissue inflammation, and reduced pro-inflammatory cytokine production late in infection. Splenocytes from BHIM-Ft sodB-immunized mice contained more CD4+, effector, memory T-cells, and were more effective at limiting intracellular replication of Ft LVS in vitro. Concurrent with enhanced killing of Ft LVS, BHIM-Ft sodB-immune splenocytes produced significantly higher levels of IFN-γ and IL-17A cytokines than their MHM-Ft sodB-immunized counterparts indicating development of a more effective T cell memory response when immunizing mice with BHIM-Ft.

Abstract LB-167: Novel LSD-1 inhibitor validation in new established clinically PFA patient-derived orthotopic xenograft (PDOX) models of ependymoma

Abstract Background: Ependymoma (EPN) is one of the most malignant tumors and surgical resection is the mainstay of treatment followed by re-irradiation and chemotherapy. However the benefit of this modality have not been clearly demonstrated. Posterior fossa A (PFA) EPN have the worst prognosis and driven by epigenetic changes, suggesting targeting epigenetic changes in PFA EPN can potentially be effective. The limited availability of a reliable translational platform has hampered efforts to understand tumor biology and the preclinical and clinical trials for EPN. In this study, we established a series of orthotopic EPN xenograft mouse models from fresh surgical specimens. We also examined the therapeutic efficacy of SYC-836, a novel LSD-1 inhibitor developed at Baylor College of Medicine, both in vitro and in vivo in these PDOX models. Material and Methods: Clinically and pathologically available PFA EPN samples were obtained from patients who had treatment in Texas Children's Hospital. Parts of the surgical samples were dissociated into single cells and implanted into the cerebellum of SCID mice (1x105 cells /2 uL FBS). Once tumorigenicity confirmed, xenograft tumor would be serially subtransplanted in mouse brains and be cultured in vitro to develop the permanent cell line. To test if xenograft tumors precisely reflect morphologic and pathologic characteristics of their parental tumors, we compared histology, and several key parameters in invasiveness, growth rate and microvessel density by IHC. To examine in vitro anti-tumor activities, primary cultured cells from an established PDOX model of PFA EPN (4423EPN) were subjected to SYC-836 at various concentrations (0-25uM). Cell viability was measured at 5 different time points over 14 days. To validate the drug's in vivo efficacy, two established PFA EPN PDOX models, 4423EPN and 2002EPN, were utilized. 40 mice per model were implanted and divided into 4 treatment groups: 1) control , 2) radiation/standard therapy (2 Gy XRT x 5 days), 3) SYC-836 only (15mg/kg IP x 28 days), and 4) combination (radiation + SYC-836 per regimen above). Animal survival times were analyzed using log rank analysis. Results: By now we have successfully developed 6 intracerebellum xenografts. It has since been serially subtransplanted in mouse for more than 3 generations and can be cryopreserved for long-term maintenance of tumorigenicity. We showed xenograft tumors precisely replicated morphologic and pathologic characteristics of their parental tumors. SYC-836 demonstrated effective cell killing in vitro in both time- and dose-dependent manner. IC50 was ~7.5uM. In vivo experiment was completed in 2 PFA EPN PDOX models. There were no survival benefit with either XRT only or SYC-836 only compared to control group; however, the combination treatment lead to significant improvement in animal survival. SYC-836 was well tolerated in mice. Conclusion: We have successfully established a series of clinically orthotopic EPN xenograft models and they well-matched with those of parental EPNs both preclinically and clinically. Our data showed that combining SYC-836 with radiation synergistically prolongs animal survival significantly, suggesting that SYC-836 may have a role in the clinical setting by either reducing radiation dosages, or be a potential adjuvant agent to other chemotherapy drugs in treatment for EPN. Citation Format: Huiyuan Zhang, Sibo Zhao, Yuchen Du, Lin Qi, Frank K. Braun, Mari Kogiso, Holly B. Lindsay, Sarah G. Injac, Laszlo Perlaky, Patricia Baxter, Jack M. Su, Xiao-Nan Li. Novel LSD-1 inhibitor validation in new established clinically PFA patient-derived orthotopic xenograft (PDOX) models of ependymoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-167.

Abstract LB-069: Identification of personalized active agents in pediatric medulloblastoma through high-throughput drug screening in matching pairs of patient derived orthotopic xenograft neurosphere and monolayer cells

Abstract Objective: Glioblastoma and Medulloblastoma are deadliest brain tumors both in adults and children. While next-generation sequencing rapidly identifies whole exome/genome gene mutations, the efficacy of targeting mutated genes/pathways has not been systematically analyzed. Additionally, it remains unknown if neurospheres (enriched with cancer stem cells) and non-stem monolayer tumor cells will respond equally to the same target therapies. Methods: Matching pairs of neurosphere and monolayer cell cultures from 4 molecularly characterized PDOX mouse models established from distinct clinical stages of pediatric GBM and MB were treated with 7,913 drugs (mostly in clinical use or trials) at 4 doses (0-10 uM) for 3 and 7 days. Dose-response curves and active area under the curve were constructed to identify active drugs and correlate with model-specific gene mutations. Results: Relatively long-term treatment (7 day screening) was achieved for high-throughput drug screening and revealed time- and dose-dependent effects in most of the 4,629 compounds that were active in at least one culture at one time point. When gene mutations were druggable, not all inhibitors of the same family were active. In IC-4687GBM (a treatment-naïve GBM), high NF1 mutation (&amp;gt;76-99% allele) frequency was found in patient tumor, xenografts, and cultured cells, but only 3/17 MEK inhibitors were active in neurospheres and 2/17 in monolayer cells (GDC0980 and PI-103 were active in both) on day 7. In IC-R0315GBM (from an autopsied terminal GBM) that carried PI3KCA mutation (allele frequency 22-27%), 5/33 PI3K inhibitors were active in neurospheres and 8/33 in monolayer cells (only Tremetinib was active in both) after 7 days. In IC-3752GBM (recurrent GBM) and ICb-1127AA (radiation-induced anaplastic astrocytoma), no druggable mutations were detected. The number of active drugs on day 7 was 366 in IC-4768GBM, 406 in IC-3752GBM, 284 in IC-R0315GBM, and 305 in ICb-1277AA. When the 4 matching pairs of neurospheres and monolayers were compared, the agents active in both cultures ranged from 36% to 60%, active only in neurosphere from 10.3% to 25%, and active only in monolayer cells from 14.8% to 53%. The response of MB to the drugs is on the way for analysis. Conclusion: We showed that long-term treatment is feasible for high-throughput drug screening. Targeting druggable mutations can be achieved but only by a fraction of specific agents. Neurospheres and monolayer cells do not always respond equally toward the same drugs, and effective targeting of both subpopulations is needed to generate prolonged animal survival times. Citation Format: Lin Qi, Yuchen Du, Goeun Bae, Mari Mari, Frank Braun, Holly Lindsay, Huiyuan Zhang, Sibo Zhao, Sarah Injac, Patricia Baxter, Jack Su, Michael Mancini, Oliver Hampton, William Parsons, Mural Chintagumpala, Clifford Stephan, Peter Davies, Xiaonan Li. Identification of personalized active agents in pediatric medulloblastoma through high-throughput drug screening in matching pairs of patient derived orthotopic xenograft neurosphere and monolayer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-069.

The Improved Delivery to Breast Cancer Based on a Novel Nanocarrier Modified with High-Affinity Peptides Discovered by Phage Display.

Ligand-targeted nanosystems have the potential to realize site-specific tumor therapy and alleviate unwanted side effects of many chemotherapeutic agents, and one of the most key issues seems to be the construction of an effective nanocarrier. Based on different processes of phage display techniques, 38 cell-binding peptides and 32 cell-internalizing peptides are discovered. Four of these ligand peptides [FIPFDPMSMRWE (FIP), NASSFPTNSRWA (NAS), GLHTSATNLYLH (GLH), and ALAVAPSRWWNE (ALA), respectively] exhibit high affinity to MCF7 human breast cancer cells. Among them, NAS and ALA are reported for the first time, whose affinities are 20.6 and 76.3 times that of the random peptide control, respectively. Both NAS and ALA modifications to doxorubicin-loaded lipid nanosytems [LP(DOX)] show stronger tumor inhibition, longer animal survival time, and less body weight loss, compared to unmodified or control peptide modified nanosystems, on an MCF7 tumor-bearing mouse model. In conclusion, the cell-binding peptide NAS and cell-internalizing peptide ALA can be used for ligand-targeted delivery of antitumor drugs. It seems that the in vivo antitumor effect of these ligand-targeted nanosystems is closely related to their ligand-cell affinity, but fairly tolerant of the ligand types.

A highly potent CDK4/6 inhibitor was rationally designed to overcome blood brain barrier in gliobastoma therapy

Glioblastoma multiforme (GBM) is the most common and deadliest of malignant brain tumors in adults. Disease development is associated with dysregulation of the cyclin D-CDK4/6-INK4-Rb pathway, resulting in increased proliferation; thus, CDK4/6 kinase inhibitors are promising candidates for GBM treatment. The recently developed CDK4/6 inhibitors, palbociclib, ribociclib, and abemaciclib, are effective in subcutaneous glioma models, but their blood-brain barrier (BBB) permeability is poor, limiting drug delivery to the central nervous system. Here, we designed and synthesized a series of novel CDK4/6 inhibitors with favorable BBB permeability for the treatment of GBM. Compound 11 exhibited a favorable pharmacological profile and significant penetration of the BBB with the Kp value of 4.10 and the Kp,uu value of 0.23 in mice after an oral dose of 10 mg/kg. IC50 values for CDK4/cyclin D1 and CDK6/cyclin D3 were 3 nM and 1 nM, respectively. In vivo studies with an orthotopic xenograft mouse model of GBM showed that 11 had tumor growth inhibition values ranging from 62% to 99% for doses ranging from 3.125 to 50 mg/kg, and no significant body weight loss was observed. The increase in life span based on the median survival time of vehicle-treated animals in mice administered a dose of 50 mg/kg was significant at 162% (p < 0.0001). These results suggest that compound 11 is a promising candidate for further investigation as an effective drug for the treatment of GBM.

The earlier, the better: the effects of different administration timepoints of sorafenib in suppressing the carcinogenesis of VEGF in rats

PurposeTo investigate the optimal starting time point of sorafenib therapy in suppressing the tumor-promoting effects of VEGF up-regulation, which is frequently found after local therapy in clinical practice.MethodsVEGF was intravenously injected to imitate the evaluated expression after local tumor therapy, such as TACE. A total of 40 SD rats bearing hepatic tumors were randomly divided into four groups and sorafenib was administered at different timepoints: (A) control group: VEGF injection only; (B) initiating sorafenib 72 h prior to VEGF injection; (C) initiating sorafenib simultaneously with VEGF injection; (D) initiating sorafenib 72 h post-VEGF injection. The rate of tumor growth, median survival time, expression of VEGF, and microvessel density (MVD), as determined by immunohistochemical (IHC) examination, were compared.ResultsThe results revealed that the tumor size and median survival time were significantly different between the three sorafenib groups compared to the control group (p < 0.05). Median survival times were 19.6 ± 1.78, 31.2 ± 6.99, 27.4 ± 4.9, and 26.5 ± 4.6 days in group A, B, C, and D, respectively. Furthermore, there was a difference in statistical significance between the two sorafenib groups B and D (p = 0.04). Tumors were collected for HE staining and IHC examination. The expression levels of VEGF in B, C, and D were 42.8 ± 7.96, 71.9 ± 15.73, and 73.6 ± 13.73, and all of them were significantly lower than that in the control group (88.3 ± 13.61). Furthermore, the level of MVD was 109.2 ± 8.98 in the control group, which was significantly higher than in the three sorafenib groups (45.7 ± 16.92, 77.1 ± 16.29, and 93.6 ± 12.87, all p < 0.05).ConclusionsAccording to our results, the most suitable regimen for the administration of sorafenib is before the increased expression of VEGF, which showed a potential advantage for controlling the tumor growth and prolonging the survival time of test animal via inhibiting VEGF-receptor expression through the bifunction of VEGF, and the reduction of tumor angiogenesis.

In vivo antitumor potential of extracts from different parts of Bauhinia variegata linn. Against b16f10 melanoma tumour model in c57bl/6 mice

BackgroundMelanoma is a metastatic type of skin cancer that is difficult to treat and the majority of efforts are directed to the design of new drugs. Medicinal Plants have been the primary source of medicines since life on earth; more than 50% of existing cancer treatments is derived from plants. Bauhinia variegata is well-known medicinal plant used from the ancient era to till date for their medicinal values. Scientific literatures have not documented any evidence of the antitumour potential of Bauhinia variegata against B16F10 melanoma tumor model in C57BL mice. The present investigation was undertaken to explore the antitumour activity of Leaf, stem bark and flower extract of Bauhinia variegata against B16F10 melanoma tumour model in C57BL mice.MethodsHydro-methanolic extract prepared from the leaf, stem bark and flower of Bauhinia variegata were assessed for their antitumor activity. The extracts at doses of 500 and 750 mg/kg b.wt. were given orally along with cyclophosphamide (chemotherapeutic drug) for 40 days for exploring antitumor activity against melanoma tumor (B16F10) in C57BL mice. Inhibition of tumor growth, increase in survival time of animal with treatment, histopathological studies and antioxidant parameter were determined.ResultsThe Present investigation showed significant effect of the B. variegata L. in preventing melanoma tumor by B16F10 cell line in C57BL/6 mice. As compared with the tumour control group, the remarkable results especially in the group which received B. variegata extract and cyclophosphamide together were obtained for all of the measured parameters. Dose dependent response was observed in tumor volume, inhibition rate, life span time and antioxidant parameter of extracts. Combination treatment of cyclophosphamide and B. variegata extracts showed more pronounced effect.ConclusionsThese findings suggest that B. variegata hydromethanolic extract may contain bioactive compounds of potential therapeutic significance which are relatively safe from toxic effects, and can compromise the medicinal use of this plant in folk medicine.

The Combined Antitumor Effects of 125I Radioactive Particle Implantation and Cytokine-Induced Killer Cell Therapy on Xenograft Hepatocellular Carcinoma in a Mouse Model.

The combination of radiotherapy and immunotherapy has shown great promise in eradicating tumors. For example, 125I radioactive particle implantation and cytokine-induced killer cell therapies have demonstrated efficacy in treating hepatocellular carcinoma. However, the mechanism of this combination therapy remains unknown. In this study, we utilized cytokine-induced killer cells obtained from human peripheral blood mononuclear cells along with 125I radioactive particle implantation to treat subcutaneous hepatocellular carcinoma xenograft tumors in BALB/c nude mice. The effects of combination therapy on tumor growth, tumor cell apoptosis and proliferation, animal survival, and immune indexes were then assessed. The results indicated that 125I radioactive particle implantation combined with cytokine-induced killer cells shows a much greater antitumor therapeutic effect than either of the therapies alone when compared to control treatments. Mice treated with a combination of radiotherapy and immunotherapy displayed significantly reduced tumor growth. 125I radioactive particle implantation upregulated the expression of major histocompatibility complex (MHC) class I chain-related gene A in hepatocellular carcinoma cells and enhanced cytokine-induced killer cell–mediated apoptosis through activation of caspase-3. Furthermore, cytokine-induced killer cells supplied immune substrates to induce a strong immune response after 125I radioactive particle implantation therapy. In conclusion, 125I radioactive particle implantation combined with cytokine-induced killer cell therapy significantly inhibits the growth of human hepatocellular carcinoma cells in vivo and improves animal survival times through mutual promotion of antitumor immunity, presenting a promising therapy for hepatocellular carcinoma.

Polymer Cancerostatics Targeted with an Antibody Fragment Bound via a Coiled Coil Motif: In Vivo Therapeutic Efficacy against Murine BCL1 Leukemia.

A BCL1 leukemia-cell-targeted polymer-drug conjugate with a narrow molecular weight distribution consisting of an N-(2-hydroxypropyl)methacrylamide copolymer carrier and the anticancer drug pirarubicin is prepared by controlled radical copolymerization followed by metal-free click chemistry. A targeting recombinant single chain antibody fragment (scFv) derived from a B1 monoclonal antibody is attached noncovalently to the polymer carrier via a coiled coil interaction between two complementary peptides. Two pairs of coiled coil forming peptides (abbreviated KEK/EKE and KSK/ESE) are used as linkers between the polymer-pirarubicin conjugate and the targeting protein. The targeted polymer conjugate with the coiled coil linker KSK/ESE exhibits 4× better cell binding activity and 2× higher cytotoxicity in vitro compared with the other conjugate. Treatment of mice with established BCL1 leukemia using the scFv-targeted polymer conjugate leads to a markedly prolonged survival time of the experimental animals compared with the treatment using the free drug and the nontargeted polymer-pirarubicin conjugate.

PH-Sensitive star-shaped poly(glutamic acid) with a porphyrin core as highly efficient photosensitizers for photodynamic therapy and molecular imaging

Recent efforts to develop tumor-targeted photodynamic therapy (PDT) photosensitizers (PSs) have greatly advanced the potential of PDT in cancer therapy, although complete eradication of tumor cells by PDT alone remains challenging. As a way to improve PDT efficacy, we report a new combinatory PDT therapy technique that specifically targets multilayers of cells. Simply mixing different PDT PSs, even those that target distinct receptors (this may still lead to similar cell-killing pathways), may not achieve ideal therapeutic outcomes. Instead, significantly improved outcomes likely require synergistic therapies that target various cellular pathways. In this study, we target two proteins upregulated in cancers: the cannabinoid CB2 receptor (CB2R, a G-protein coupled receptor) and translocator protein (TSPO, a mitochondria membrane receptor). We found that the CB2R-targeted PS, IR700DX-mbc94, triggered necrotic cell death upon light irradiation, whereas PDT with the TSPO-targeted IR700DX-6T agent led to apoptotic cell death. Both PSs significantly inhibited tumor growth in vivo in a target-specific manner. As expected, the combined CB2R- and TSPO-PDT resulted in enhanced cell killing efficacy and tumor inhibition with lower drug dose. The median survival time of animals with multilayer PDT treatment was extended by as much as 2.8-fold over single PDT treatment. Overall, multilayer PDT provides new opportunities to treat cancers with high efficacy and low side effects.Photodynamic therapy (PDT) is increasingly used as a minimally invasive, controllable and effective therapeutic procedure for cancer treatment. However, complete eradication of tumor cells by PDT alone remains challenging. In this study, we investigate the potential of multilayer PDT in cancer treatment with high efficacy and low side effects. Through PDT targeting two cancer biomarkers located at distinct subcellular localizations, remarkable synergistic effects in cancer cell killing and tumor inhibition were observed in both in vitro and in vivo experiments. This strategy may be widely applied to treat various cancer types by using strategically designed PDT photosensitizers that target corresponding upregulated receptors at tactical subcellular localization.