ABSTRACT
To overcome C797S mutation, the latest and most common resistance mechanism in the clinical treatment of third-generation EGFR inhibitor, a novel series of substituted 6-(2-aminopyrimidine)-indole derivatives were designed and synthesized. Through the structure-activity relationship (SAR) study, compound 11eg was identified as a novel and potent EGFR L858R/T790M/C797S inhibitor (IC50 = 0.053 µM) but had a weak effect on EGFRWT (IC50 = 1.05 µM). 11eg significantly inhibited the proliferation of the non-small cell lung cancer (NSCLC) cells harboring EGFRL858R/T790M/C797S with an IC50 of 0.052 µM. 11eg also showed potent inhibitory activity against other NSCLC cell lines harboring main EGFR mutants. Furthermore, 11eg exhibited much superior activity in arresting cell cycle and inducing apoptosis of NSCLC cells with mutant EGFRC797S. It blocked cellular EGFR signaling. Importantly, 11eg markedly suppressed the tumor growth in in vivo xenograft mouse model with good safety. Additionally, 11eg displayed good microsomal stability. These results demonstrated the potential of 11eg with novel scaffold as a promising lead compound targeting EGFRC797S to guide in-depth structural optimization.
ABSTRACT
In this study, we identified a newly synthesized compound 7o with potent inhibition on EGFR primary mutants (L858R, Del19) and drug-resistant mutant T790M with nanomolar IC50 values. 7o showed strong antiproliferative effects against EGFR mutant-driven non-small cell lung cancer (NSCLC) cells such as H1975, PC-9 and HCC827, over cells expressing EGFRWT. Molecular docking was performed to investigate the possible binding modes of 7o inside the binding site of EGFRL858R/T790M and EGFRWT. Analysis of cell cycle evidenced that 7o induced cell cycle arrest in G1 phases in the EGFR mutant cells, H1975 and PC-9, which resulted in decreased S-phase populations. Moreover, compound 7o induced cancer cell apoptosis in in vitro assays. In addition, 7o inhibited cellular phosphorylation of EGFR. In vivo, oral administration of 7o caused rapid tumor regression in H1975 xenograft model. Therefore, 7o might deserve further optimization as cancer treatment agent for EGFR mutant-driven NSCLC.
Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Humans , Lung Neoplasms/metabolism , Carcinoma, Non-Small-Cell Lung/metabolism , ErbB Receptors , Molecular Docking Simulation , Cell Proliferation , Protein Kinase Inhibitors , Mutation , Cell Line, Tumor , Drug Resistance, NeoplasmABSTRACT
Efforts toward finding potent CDK4 inhibitor for cancer therapy, a series of fluorine substituted pyrrolo[2,3-d]pyrimidine derivatives were designed, synthesized, and evaluated. Among them, the optimal lead compound 18i was discovered with potent activity against CDK4 at the nanomolar level (IC50 = 2.5 nM) and exquisite selectivity which demonstrated only modest activity against 3 out of the 394 protein kinases. 18i exhibited a much greater in vitro antiproliferative activity against several human cancer cell lines than that of the approved drug ribociclib. Further mechanism studies revealed that 18i effectively stimulated cancer cell cycle arrest in G1 phase and induced tumor cell apoptosis. In the comparison of in vivo therapeutic effects in xenograft mouse models of breast cancer, oral administration of 18i showed a significantly better degree of inhibitory effect to ribociclib without obvious toxicity. All of the results indicated that 18i could be a promising CDK4 inhibitor for treating malignancies.
Subject(s)
Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/therapeutic use , Cell Survival/drug effects , Cyclin-Dependent Kinase 4/antagonists & inhibitors , Drug Design , Animals , Antineoplastic Agents/chemistry , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , Cyclin-Dependent Kinase 4/genetics , Cyclin-Dependent Kinase 4/metabolism , Gene Expression Regulation, Neoplastic/drug effects , Humans , Mice , Mice, Nude , Molecular Structure , Neoplasms, Experimental/drug therapy , Phylogeny , Random Allocation , Signal TransductionABSTRACT
A novel series of pyrimidine-benzotriazole derivatives have been synthesized and evaluated for their anticancer activity against human solid tumor cell lines. The most promising molecule 12O was identified for its excellent antiproliferative activities, especially against the SiHa cell line with IC50 value as 0.009 µM. Kinase inhibition assay assessed 12O was a potential multi-kinase inhibitor, which possessed potent inhibitory activities against cyclin-dependent kinases (CDKs) and fms-like tyrosine kinase (FLT) with IC50 values in the nanomolar range. Molecular docking studies illustrated that the introduction of triazole moiety in 12O was critical for CDKs inhibition. In addition, 12O inhibited cancer cell proliferation, colony-formation, and cell cycle progression and provoked apoptotic death in vitro. In an SiHa xenograft mouse model, a once-daily dose of compound 12O at 20 mg/kg significantly suppressed the tumor growth without obvious toxicity. Taken together, 12O provided valuable guide for further structural optimization for CDKs and FLT inhibitors.
Subject(s)
Antineoplastic Agents/pharmacology , Cyclin-Dependent Kinases/antagonists & inhibitors , Neoplasms/drug therapy , Protein Kinase Inhibitors/pharmacology , Vascular Endothelial Growth Factor Receptor-1/antagonists & inhibitors , Administration, Oral , Animals , Antineoplastic Agents/administration & dosage , Cell Line, Tumor , Cell Proliferation , Drug Design , Drug Screening Assays, Antitumor , Female , HeLa Cells , Humans , Inhibitory Concentration 50 , MCF-7 Cells , Mice , Mice, Inbred BALB C , Mice, Nude , Molecular Docking Simulation , Neoplasm Transplantation , Protein Kinase Inhibitors/administration & dosage , Pyrimidines/chemistryABSTRACT
A novel series of cyclin-dependent kinases (CDKs) inhibitors, which play critical roles in the cell cycle control and regulation of cell transcription, were synthesised. A systematic study of enzymatic and cellular assays led to the identification of compound X22 with a nanomolar potency against CDK4 and CDK9 and potent antiproliferative activities against a panel of tumour cell lines. X22 could induce cell cycle arrest and cell apoptosis in cancer cell lines. X22 dose-dependently inhibits signalling pathways downstream of CDKs in cancer cells. In vivo antitumor activity assays, oral administration of X22 led to significant tumour regression in mouse model without obvious toxicity. Superior anti-cancer efficacy in vitro and in vivo of X22 demonstrated combined depletion of cell cycle and transcriptional CDK all contributed to antitumor activity. Taken together, concomitant inhibition of cell cycle and transcriptional CDK activities provided valuable guide for further structural optimisation.
Subject(s)
Antineoplastic Agents/pharmacology , Cell Cycle/drug effects , Cyclin-Dependent Kinases/antagonists & inhibitors , Neoplasms/drug therapy , Protein Kinase Inhibitors/pharmacology , Transcription, Genetic/drug effects , Animals , Antineoplastic Agents/therapeutic use , Cell Line, Tumor , Drug Screening Assays, Antitumor , Female , Humans , Mice , Mice, Inbred BALB C , Neoplasms/enzymology , Neoplasms/pathology , Protein Kinase Inhibitors/chemistry , Protein Kinase Inhibitors/therapeutic use , Structure-Activity Relationship , Xenograft Model Antitumor AssaysABSTRACT
BACKGROUND/PURPOSE: To evaluate the ability of quadruple Taqman probe real-time PCR to the detection of vanA, vanB and vanM in enterococcal isolates. METHODS: A total of 343 strains, including 253 vancomycin-resistant enterococcus (VRE) strains and 90 non-VRE strains, were tested by both quadruple Taqman probe real-time PCR and gel-based PCR assay. RESULTS: When differentiating among three genotypes of vanA, vanB and vanM in VRE strains, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), diagnostic accuracy and consistency of the quadruple Taqman probe real-time PCR were all 100%. Minimum. Inhibitory concentration (MIC) results showed that there was a wide MIC range of vancomycin and teicoplanin for the strains that harboring vanA/vanM gene respectively or harboring vanA and vanM genes simultaneously. However, the VRE strains with vanB genotype all were sensitive to teicoplanin. CONCLUSION: Considering the excellent PPV and low NPV, real-time PCR would be useful to monitor VRE-colonized or infected patients. However, further evaluation of the assay's performance in the clinical specimens is required, especially when considering that high level of PCR inhibitors present in these samples.
Subject(s)
Bacterial Proteins/genetics , Bacterial Proteins/isolation & purification , Carbon-Oxygen Ligases/genetics , Carbon-Oxygen Ligases/isolation & purification , Real-Time Polymerase Chain Reaction/methods , Acinetobacter baumannii , Drug Resistance, Microbial/genetics , Enterococcus/drug effects , Enterococcus/genetics , Enterococcus faecium/genetics , Genotype , Klebsiella pneumoniae/genetics , Microbial Sensitivity Tests , Pseudomonas aeruginosa , Staphylococcus aureus/genetics , Teicoplanin/pharmacology , Vancomycin/pharmacology , Vancomycin-Resistant Enterococci/geneticsABSTRACT
A series of novel, highly potent, selective CDK9 inhibitors with cancer stem cells (CSCs) inhibition activity were designed and synthesized for non-small-cell lung cancer (NSCLC) therapy. Structure-activity relationship analysis based on enzymatic and cellular activities led to the discovery of a promising inhibitor 21e. 21e potently inhibited CDK9 with IC50 value of 11â¯nM and suppressed the stemness properties of NSCLC effectively. It could decrease the stemness phenotypes of NSCLC cells, including tumor sphere formation, side-population and stemness markers abundance. 21e displayed good selectivity over the CDK family kinases and kinase profiling assay against 381 kinases. In addition, 21e inhibited cell proliferation, colony-formation, and cell cycle progression and induced apoptosis in NSCLC. In H1299 xenograft mouse model, a once-daily dose of compound 21e at 20â¯mg/kg significantly suppressed the tumor growth without obvious toxicity. Studies of mechanisms of action indicated that 21e efficiently inhibited CDK9 signaling pathway and stemness both in vitro and in vivo. Collectively, 21e as a novel CDK9 inhibitor with CSCs inhibition properties could be a promising agent for the treatment of NSCLC.
Subject(s)
Antineoplastic Agents/pharmacology , Carcinoma, Non-Small-Cell Lung/drug therapy , Cyclin-Dependent Kinase 9/antagonists & inhibitors , Isothiocyanates/pharmacology , Lung Neoplasms/drug therapy , Protein Kinase Inhibitors/pharmacology , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/chemistry , Apoptosis/drug effects , Carcinoma, Non-Small-Cell Lung/metabolism , Cell Proliferation/drug effects , Cyclin-Dependent Kinase 9/metabolism , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Humans , Isothiocyanates/chemical synthesis , Isothiocyanates/chemistry , Lung Neoplasms/metabolism , Molecular Structure , Protein Kinase Inhibitors/chemical synthesis , Protein Kinase Inhibitors/chemistry , Structure-Activity Relationship , Sulfoxides , Tumor Cells, CulturedABSTRACT
In this paper, to achieve the targeted ability of anti-tumor drug doxorubicin (DOX), enhance the treatment effect and reduce the side effect, a novel pH-sensitive and charge-convertible prodrug nanogel was prepared. Firstly, cis-aconitic anhydride-doxorubicin prodrug (CAD) and Pluronic F127-chitosan-CAD (F127-CS-CAD) conjugates were synthesized. Then the DOX loaded polyion complex micelles (F127-CS-CAD/CAD) were prepared by self-assembling, thus CAD was incorporated into micelles via electrostatic interactions between electronegative CAD and positively charged F127-CS-CAD and hydrophobic interactions. Finally a pH-responsive charge-convertible copolymer, folic acid modified gelatin (Gel-FA) was shielded on the surface of micelles and the Gel-FA/F127-CS-CAD/CAD nanogel was formed, the charge-convertible capability was evaluated through changes of the morphology and Zeta potential under different pH value environment by transmission electron microscopy (TEM) and Zeta potential analyzer. And in vitro pH-dependent and two-phase drug release from nanogel was also evaluated. In vitro anti-tumor activity of Gel-FA/F127-CS-CAD/CAD nanogel was performed on HeLa cells and HepG2 cells to prove the strong cell toxicity of nanogels. Finally, the in vivo safety experiments showed that the nanogel achieved the reducing the toxic side effects of DOX significantly.
Subject(s)
Doxorubicin/chemistry , Nanoparticles/chemistry , Prodrugs/chemistry , HeLa Cells , Humans , Hydrogen-Ion Concentration , Micelles , Microscopy, Electron, TransmissionABSTRACT
Despite initial progress in preclinical models, most known histone deacetylase inhibitors (HDACis) used as a single agent have failed to show clinical benefits in nearly all types of solid tumours. Hence, the efficacy of HDACis in solid tumours remains uncertain. Herein, we developed a hybrid HDAC inhibitor that sensitized solid tumours to HDAC-targeted treatment. Methods: A hybrid molecule, Roxyl-zhc-84 was designed and synthesized with novel architecture. The pharmacokinetics and toxicity of Roxyl-zhc-84 were analysed. The antitumour effects of Roxyl-zhc-84 on solid tumours were investigated by assessing cell growth, apoptosis and cell cycle in vitro and in three in vivo mouse models and compared to those of corresponding control inhibitors alone or in combination. Gene set enrichment analysis was performed, and relevant JAK1-STAT3-BCL2 signalling was identified in vitro and in vivo in mechanistic studies. Results: Roxyl-zhc-84 showed excellent pharmacokinetics and low toxicity. The novel hybrid inhibitor Roxyl-zhc-84 induced cell apoptosis and G1-phase arrest in breast cancer and ovarian cancer cell lines. In three mouse models, oral administration of Roxyl-zhc-84 led to significant tumour regression without obvious toxicity. Moreover, Roxyl-zhc-84 dramatically improved the limited response of traditional HDAC inhibitors in solid tumours via overcoming JAK1-STAT3-BCL2-mediated drug resistance. Roxyl-zhc-84 treatment exhibited vastly superior efficacy than the combination of HDAC and JAK1 inhibitors both in vitro and in vivo. Conclusion: Concurrent inhibition of HDAC and CDK using Roxyl-zhc-84 with additional JAK1 targeting resolved the limited response of traditional HDAC inhibitors in solid tumours via overcoming JAK1-STAT3-BCL2-mediated drug resistance, providing a rational multi-target treatment to sensitize solid tumours to HDACi therapy.
Subject(s)
Antineoplastic Agents/administration & dosage , Breast Neoplasms/drug therapy , Histone Deacetylase Inhibitors/administration & dosage , Ovarian Neoplasms/drug therapy , Signal Transduction/drug effects , Animals , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/toxicity , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Proliferation/drug effects , Disease Models, Animal , Female , Gene Expression Profiling , Janus Kinase 1/metabolism , Mice , Models, Theoretical , Proto-Oncogene Proteins c-bcl-2/metabolism , STAT3 Transcription Factor/metabolism , Treatment OutcomeABSTRACT
In order to solve the problem of the short lifespan of the neural electrode caused by micro motion, this study designed a novel neural electrode based on lumped compliance compliant mechanism to control different modes of micro-motion in a more effective way. According to the mathematical modeling of the novel neural electrode, the equivalent bending stiffness and equivalent tensile (compression) stiffness were calculated. The results of the finite element analysis based on the mathematical modeling revealed that the novel neural electrode showed excellent micro-motion-attenuation capability. The static analysis results showed that the novel design dramatically reduced the maximum displacement of the brain in 51% and the maximum stress in 41% under longitudinal micro-motion environment. It also effectively reduced the 5.1% maximum stress while maintaining the maximum displacement under lateral micro-motion environment. The experimental results based on the tissue injury evaluation system also confirmed that the novel electrode is more effective in micro-motion attenuation than the reference one. In detail, the strain of the brain tissue caused by the implantation of the neural electrode was decreased by 1.26 to 27.84% at the insertion depth of 3 mm, and 0.522 to 17.24% at the insertion depth of 4.5 mm, which has convinced the effectiveness of the design. Graphical abstract The schematic of the novel neural electrode and evaluationsystem of tissue injury.
Subject(s)
Electrodes, Implanted , Motion , Neurons/physiology , Brain/physiology , Computer Simulation , Finite Element Analysis , Imaging, Three-Dimensional , Silicon/chemistry , Stress, MechanicalABSTRACT
On the basis of novel pyrazino[2,3-c]quinolin-2(1H)-one scaffold, we designed and identified a highly selective, potent and oral mTOR inhibitor, 9m. Compound 9m showed low nanomolar activity against mTOR (IC50 = 7 nM) and greater selectivity over the related PIKK family kinases, which demonstrated only modest activity against 3 out of the 409 protein kinases. In vitro assays, compound 9m exhibited high potency against human breast and cervical cancer cells and induced tumor cell cycle arrest and autophagy. 9m inhibited cellular phosphorylation of mTORC1 (pS6 and p4E-BP1) and mTORC2 (pAKT (S473)) substrates. In T-47D xenograft mouse model, oral administration of compound 9m led to significant tumor regression without obvious toxicity. In addition, this compound showed good pharmacokinetics. Collectively, due to its high potency and selectivity, compound 9m could be used as a mTOR drug candidate.
Subject(s)
Antineoplastic Agents/administration & dosage , Antineoplastic Agents/pharmacology , Autophagy/drug effects , Protein Kinase Inhibitors/administration & dosage , Protein Kinase Inhibitors/pharmacology , TOR Serine-Threonine Kinases/antagonists & inhibitors , Administration, Oral , Cell Line, Tumor , Humans , Models, Molecular , Phosphorylation/drug effects , Protein Domains , TOR Serine-Threonine Kinases/chemistryABSTRACT
Nanogel-based multifunctional drug delivery systems, especially hybrid nanogels and multicompartment nanogels have drawn more and more extensive attention from the researchers in pharmacy because it can result in achieving a superior functionality through the synergistic property enhancement of each component. The unique hybrid and compartmentalized structures provide the great potential for co-delivery of multiple agents even the multiple agents with different physicochemical properties. Otherwise the hybrid nanogel encapsulating optical and magnetic resonance imaging contrast can be utilized in imaging technique for disease diagnosis. More importantly through nanogel-based multifunctional drug delivery systems the stimuli-responsive features might be easily employed for the design of targeted release of drug. This review summarizes the construction of diverse hybrid nanogels and multicompartment nanogels. The application in co-delivery of multiple agents and imaging agents for diagnosis as well as the application in the design of stimuli-responsive multifunctional nanogels as drug delivery are also reviewed and discussed. The future prospects in application of multifunctional nanogels will be also discussed in this review.
Subject(s)
Drug Delivery Systems/methods , Magnetic Resonance Imaging/methods , Nanostructures , Tomography, Optical Coherence/methods , Animals , Gels/chemical synthesis , Gels/chemistry , Gels/therapeutic use , Humans , Nanostructures/chemistry , Nanostructures/therapeutic useABSTRACT
Single drug therapy that leads to the multidrug resistance of cancer cells and severe side-effect is a thing of the past. Combination therapies that affect multiple signaling pathways have been the focus of recent active research. Due to the successful development of prodrug-based nano-drug delivery systems (P-N-DDSs), their use has been extended to combination therapy as drug delivery platforms. In this review, we focus specifically on the P-N-DDSs in the field of combination therapy including the combinations of prodrugs with different chemotherapeutic agents, other therapeutic agents, nucleic acid or the combination of different types of therapy (e.g. chemotherapy and phototherapy). The relevant examples of prodrug-based nanoparticulate drug delivery strategy in combination cancer therapy from the recent literature are discussed to demonstrate the feasibilities of relevant technology.
Subject(s)
Antineoplastic Agents/administration & dosage , Drug Delivery Systems , Nanoparticles/administration & dosage , Neoplasms/therapy , Prodrugs/administration & dosage , Combined Modality Therapy , Humans , Nanoparticles/chemistryABSTRACT
PURPOSE: The application of neural probes in clinic has been challenged by probes' short lifetime when implanted into brain tissue. The primary goal is to develop an evaluation system for testing brain tissue injury induced by neural probe's insertion using microscope based digital image correlation method. METHODS: A brain tissue phantom made of silicone rubber with speckle pattern on its surface was fabricated. To obtain the optimal speckle pattern, mean intensity gradient parameter was used for quality assessment. The designed testing system consists of three modules: (a) load module for simulating neural electrode implantation process; (b) data acquisition module to capture micrographs of speckle pattern and to obtain reactive forces during the insertion of the probe; (c) postprocessing module for extracting tissue deformation information from the captured speckle patterns. On the basis of the evaluation system, the effects of probe wedge angle, insertion speed, and probe streamline on insertion induced tissue injury were investigated. RESULTS: The optimal quality speckle pattern can be attained by the following fabrication parameters: spin coating rate-1000 r/min, silicone rubber component A: silicone rubber component B: softener: graphite = 5 ml: 5 ml: 2 ml: 0.6 g. The probe wedge angle has a significant effect on tissue injury. Compared to wedge angle 40° and 20°, maximum principal strain of 60° wedge angle was increased by 40.3% and 87.5%, respectively; compared with a relatively higher speed (500 µm/s), the maximum principle strain within the tissue induced by slow insertion speed (100 µm/s) was increased by 14.3%; insertion force required by probe with convex streamline was smaller than the force of traditional probe. Based on the experimental results, a novel neural probe that has a rounded tip covered by a biodegradable silk protein coating with convex streamline was proposed, which has both lower insertion and micromotion induced tissue injury. CONCLUSIONS: The established evaluation system has provided a simulation environment for testing brain tissue injury produced by various insertion conditions. At the same time, it eliminates the adverse effect of biological factors on tissue deformation during the experiment, improving the repeatability of measurement results. As a result, the evaluation system will provide support on novel neural probe design that can reduce the acute tissue injury during the implantation of the probe.
Subject(s)
Brain Injuries/etiology , Brain Injuries/pathology , Electrodes, Implanted/adverse effects , Microscopy/methods , Artifacts , Microscopy/instrumentation , Phantoms, Imaging , Silicones , Stress, Mechanical , Time FactorsABSTRACT
A doxorubicin conjugated prodrug incorporated acid-sensitive linkage between drug and Pluronic F127-chitosan (F127-CS) polymer was successfully synthesized. Subsequently a pH-sensitive polymeric micelle system was designed based on the conjugated prodrugs (F127-CS-DOX) to co-deliver doxorubicin and paclitaxel. Paclitaxel (PTX) was physically entrapped in the hydrophobic inner core of the micelles simultaneously. The structures of conjugates were analyzed by means of (1)H NMR and UV-vis spectrum. Size distribution and morphology of the micelles were observed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The results indicated that obtained micelles had good dispersity and the diameter was between 56.3 and 403.4 nm. The loading of PTX into the micelle increased with higher DOX content. DOX and PTX release from polymeric micelles followed an acid-triggered manner. Furthermore, in vivo pharmacokinetic study also showed that the area under the plasma concentration time curve (AUC0-∞) values of PTX and DOX for PTX-loaded F127-CS-DOX micelles in rats were 3.97 and 4.38-fold higher than those for PTX plus DOX solution. These results suggested the PTX-loaded F127-CS-DOX micelles would be a promising carrier for co-delivering DOX and PTX.
Subject(s)
Doxorubicin/chemistry , Paclitaxel/chemistry , Polymers/chemistry , Animals , Hydrogen-Ion Concentration , Magnetic Resonance Spectroscopy , Male , Micelles , Microscopy, Electron, Transmission , Rats , Rats, Wistar , Spectroscopy, Fourier Transform InfraredABSTRACT
Given the multiple terrorist attacks that have occurred in recent years in China, medical rescue teams and specialized incident assessment teams have been established by the government; however, medical rescue after nuclear, biological, and chemical incidents remains challenging and is often inefficient. In the present article, problems were analyzed regarding the assessment of responder countermeasures, training of professionals and the management of emergency medical incidents related to nuclear, biological and chemical attacks. Countermeasures, the establishment of response coordination, public education, practical training and exercise, and a professional consultant team or system should be the focus of emergency medical response facilities. Moreover, the government was offered professionals who are involved in managing nuclear, biological and chemical incidents.