ABSTRACT
Doxorubicin (DOX) is an important drug for cancer treatment, but its clinical application is limited due to its toxicity and side effects. Therefore, detecting the concentration of DOX during treatment is crucial for enhancing efficacy and reducing side effects. In this study, the authors developed a biophotonic fiber sensor based on localized surface plasmon resonance (LSPR) with the multimode fiber (MMF)-four core fiber (FCF)-seven core fiber (SCF)-MMF-based direct-taper and anti-taper structures for the specific detection of DOX. Compared to other detection methods, it has the advantages of high sensitivity, low cost, and strong anti-interference ability. In this experiment, multi-walled carbon nanotubes (MWCNTs), cerium-oxide nanorods (CeO2-NRs), and gold nanoparticles (AuNPs) were immobilized on the probe surface to enhance the sensor's biocompatibility. MWCNTs and CeO2-NRs provided more binding sites for the fixation of AuNPs. By immobilizing AuNPs on the surface, the LSPR was stimulated by the evanescent field to detect DOX. The sensor surface was functionalized with DOX aptamers for specific detection, enhancing its specificity. The experiments demonstrated that within a linear detection range of 0-10 µM, the sensitivity of the sensor is 0.77 nm/µM, and the limit of detection (LoD) is 0.42 µM. Additionally, the probe's repeatability, reproducibility, stability, and selectivity were evaluated, indicating that the probe has high potential for detecting DOX during cancer treatment.
Subject(s)
Doxorubicin , Gold , Metal Nanoparticles , Surface Plasmon Resonance , Doxorubicin/pharmacology , Humans , Surface Plasmon Resonance/instrumentation , Gold/chemistry , Metal Nanoparticles/chemistry , Neoplasms/drug therapy , Nanotubes, Carbon/chemistry , Biosensing Techniques/instrumentation , Optical Fibers , Equipment Design , Antibiotics, Antineoplastic/analysis , Cerium/chemistry , Fiber Optic Technology/instrumentationABSTRACT
Nanomaterial selection is critical for photoelectrochemical (PEC) sensing. In this report, a novel cathodic photoelectrochemical (PEC) strategy was proposed for the detection of doxorubicin hydrochloride (Dox) and gentamicin sulfate (CN). The photocathode was synthesized by noncovalently coupling cadmium sulfide (CdS) to the porphyrin-derived metal-organic framework (CdS@PCN-224). This type of assembly created a pleasant interface for the combination of doxorubicin hydrochloride and gentamicin sulfate, resulting in a good CdS@PCN-224 donor-acceptor system. When compared to a single optoelectronic material, its photocurrent is enhanced by unprecedented nine times. This research could pave the way for the realization of PCN-224's enormous potential in PEC sensing.
Subject(s)
Biomimetic Materials/chemistry , Cadmium Compounds/chemistry , Doxorubicin/analysis , Gentamicins/analysis , Metal-Organic Frameworks/chemistry , Sulfides/chemistry , Anti-Bacterial Agents/analysis , Antibiotics, Antineoplastic/analysis , Biosensing Techniques , Electrochemical Techniques , Materials Testing , Molecular Structure , Particle Size , Photochemical ProcessesABSTRACT
BACKGROUND: Pressurized Intra-Peritoneal Aerosol Chemotherapy (PIPAC) is an innovative treatment against peritoneal carcinomatosis. Doxorubicin is a common intra-venous chemotherapy used for peritoneal carcinomatosis and for PIPAC. This study evaluated the impact of increased PIPAC intraperitoneal pressure on the distribution and cell penetration of doxorubicin in a sheep model. METHODS: Doxorubicin was aerosolized using PIPAC into the peritoneal cavity of 6 ewes (pre-alpes breed): N = 3 with 12 mmHg intraperitoneal pressure ("group 12") and N = 3 with 20 mmHg ("group 20"). Samples from peritoneum (N = 6), ovarian (N = 1), omentum (N = 1) and caecum (N = 1) were collected for each ewe. The number of doxorubicin positive cells was determined using the ratio between doxorubicine fluorescence-positive cell nuclei (DOXO+) over total number of DAPI positive cell nuclei (DAPI+). Penetration depth (µm) was defined as the distance between the luminal surface and the location of the deepest DOXO+ nuclei over the total number of cell nuclei that were stained with DAPI. Penetration depth (µm) was defined as the distance between the luminal surface and the location of the deepest DOXO+ nuclei. RESULTS: DOXO+ nuclei were identified in 87% of samples. All omental samples, directly localized in front of the nebulizer head, had 100% DOXO+ nuclei whereas very few nuclei were DOXO+ for caecum. Distribution patterns were not different between the two groups but penetration depth in ovary and caecum samples was significantly deeper in group 20. CONCLUSIONS: This study showed that applying a higher intra-peritoneal pressure during PIPAC treatment leads to a deeper penetration of doxorubicin in ovarian and caecum but does not affect distribution patterns.
Subject(s)
Antibiotics, Antineoplastic/pharmacokinetics , Doxorubicin/pharmacokinetics , Drug Delivery Systems/methods , Peritoneal Neoplasms/metabolism , Aerosols , Animals , Antibiotics, Antineoplastic/administration & dosage , Antibiotics, Antineoplastic/analysis , Cecum/chemistry , Cecum/metabolism , Cell Nucleus/chemistry , Doxorubicin/administration & dosage , Doxorubicin/analysis , Female , Omentum/chemistry , Omentum/metabolism , Ovary/chemistry , Ovary/metabolism , Peritoneal Neoplasms/drug therapy , Peritoneum/chemistry , Peritoneum/metabolism , Pressure , Sheep , Tissue DistributionABSTRACT
The recent development of an in vivo solid-phase microextraction (SPME) method capable of analyzing drugs and metabolic products in biofluids and living tissues holds great promise. The standard in vivo SPME protocol based on mass spectrometry is a very powerful analytical approach, but it is not practical for on-site analysis in many cases. In this paper, we present a fluorescence-based SPME method and a prototype of a portable fluorometer that is capable of quickly quantifying concentrations of the anticancer drug, doxorubicin (DOX). The instrument uses thin coated, biocompatible SPME fibers, which we have previously presented as a chemical biopsy tool for use during in vivo lung perfusion (IVLP) procedures within a hospital setting. In this research, we test SPME fibers with C8-SCX, C18, and HLB coatings with our fluorometer. The mixed-mode C8-SCX fibers showed the best sensitivity of the three and were therefore used to examine DOX extraction from perfusate solution and a homogenized lamb lung tissue. The maximum concentration of free active sites in the C8-SCX fiber and the adsorption equilibrium constant were determined to be (9.1 ± 0.3) × 10-7 mol m-2 and 420 ± 30 m3 mol-1, respectively. Finally, the detection limits for DOX extracted from buffer, perfusate, and lung tissue were 40, 100, and 3700 µg L-1, respectively.
Subject(s)
Antibiotics, Antineoplastic/analysis , Body Fluids/chemistry , Doxorubicin/analysis , Fluorometry , Lung/chemistry , Solid Phase Microextraction , Antibiotics, Antineoplastic/metabolism , Doxorubicin/metabolism , Humans , Perfusion , Solutions , Spectrometry, FluorescenceABSTRACT
The occurrence of pharmaceutical residues in surface water is raising environmental concern. To accompany the evolution of measures for natural resources protection, sensing methods enabling sensitive and rapid water quality monitoring are needed. We recently managed the parallelization of the Tethered Particle Motion (TPM), a single molecule technique, sensitive to the conformational changes of DNA. Here, we investigate the capacity of high throughput TPM (htTPM) to detect drugs that intercalate into DNA. As a proof-of-concept we analyze the htTPM signal for two DNA intercalating dyes, namely, YOYO-1 and SYTOX orange. The efficient detection of intercalating drugs is then demonstrated with doxorubicin. We further evaluate the possibility to detect carbamazepine, an antiepileptic massively prescribed and persistent in water, which had been described to interact with DNA through intercalation. Our results corroborated by other techniques show that, in fact, carbamazepine is not a DNA intercalator. The comparison of the results obtained with different aqueous buffers and solutions allows us to identify optimal conditions for the monitoring of intercalation compounds by htTPM.
Subject(s)
Antibiotics, Antineoplastic/analysis , Benzoxazoles/chemistry , DNA/chemistry , Doxorubicin/analysis , Fluorescent Dyes/chemistry , Quinolinium Compounds/chemistry , Organic Chemicals/chemistry , Water/chemistryABSTRACT
Objectives: In recent years, various formulations containing rapamycin, mainly petrolatum-based, have been tested on facial angiofibromas in tuberous sclerosis. They are often poorly tolerated due to irritation and bleeding. In addition, their effectiveness was insufficient in young adults. The objective of this study was to develop and characterise a hydro-alcoholic gel containing solubilised rapamycin. The stability of the product stored at 4°C was evaluated over 1 year. Methods: Two different 0.1% rapamycin gels were formulated with or without α-tocopherol and urea. Different methods were used to characterise the gels: HPLC, gas chromatography, pH, visual observation and optical microscopy. A physico-chemical and microbiological stability study was also conducted for 1 year at 4°C. Results: Gels were physically and microbiologically stable after 1 year at 4°C: organoleptic characteristics and pH unchanged, no significant decrease in rapamycin was observed, tocopherol droplet size was constant and rheological behaviour was not altered. Conclusions: This study describes a new gel formulation to improve skin penetration using various excipients to promote skin tolerance. This study provides, for the first time, detailed stability data for a hydro-alcoholic rapamycin gel.
Subject(s)
Angiofibroma/drug therapy , Antibiotics, Antineoplastic/chemistry , Drug Compounding/trends , Facial Neoplasms/drug therapy , Sirolimus/chemistry , Administration, Topical , Antibiotics, Antineoplastic/administration & dosage , Antibiotics, Antineoplastic/analysis , Chromatography, High Pressure Liquid/methods , Chromatography, High Pressure Liquid/trends , Drug Compounding/methods , Drug Stability , Gels , Humans , Hydrophobic and Hydrophilic Interactions , Sirolimus/administration & dosage , Sirolimus/analysis , Treatment OutcomeABSTRACT
An innovative and portable design to fabricate an electrochemical sensor based on metallic phase MoS2 (1T-MoS2) decorated with shape-dependent gold nanostructures for the determination of doxorubicin (DOX) is presented. In this context, homogenous and uniform single-crystal gold nanospheres (AuNSPs) and nanorods (AuNRDs) were firstly synthesized by seeded growth approach. Afterwards, AuNSPs and AuNRDs were anchored on 1T-MoS2 surfaces to construct the desired electrochemical sensing platform towards the DOX assay. 1T-MoS2 was exfoliated by metal intercalation process using NaK metal alloys. The structure and surface morphology of 1T-MoS2, AuNSPs, and AuNRDs were characterized by XPS, Raman, UV-vis, TEM, and SEM. The electrochemical behavior of DOX using various MoS2-based electrochemical sensors prepared on screen-printed electrode (SPE) was examined by cyclic voltammetry and adsorptive stripping differential pulse voltammetry. The electrocatalytic efficiency of AuNRDs on 1T-MoS2 was also compared with that of AuNSPs on 1T-MoS2, and it showed much better electrocatalytic activity towards the DOX. A nanocomposite prepared with AuNRDs and 1T-MoS2 on SPE (AuNRDs/1T-MoS2/SPE) exhibited a linear relationship between peak current and DOX concentration in the range 0.01-9.5 µM with a detection limit of 2.5 nM. The AuNRDs/1T-MoS2/SPE was successfully applied to the sensitive and rapid determination of DOX in spiked human serum samples with satisfactory recoveries in the range 99.2-100.8%. Graphical abstract Schematic representation of a portable design for electrochemical sensor based on shape-controlled gold nanostructures decorated on metallic phase molybdenum disulfide (1T-MoS2) towards the sensitive determination of doxorubicin.
Subject(s)
Antibiotics, Antineoplastic/analysis , Biosensing Techniques , Disulfides/chemistry , Doxorubicin/analysis , Electrochemical Techniques , Gold/chemistry , Molybdenum/chemistry , Nanostructures/chemistry , HumansABSTRACT
Doxorubicin (DOX) ranks among the most effective anticancer agents. Increasing the formation of covalent DOX-DNA interstrand cross-links can improve the anticancer activity of DOX. However, due to the low stability of the DOX-DNA cross-links to heat and alkali, DOX can be extensively lost during isolation procedures of biochemical methods, thus reducing the apparent clinical relevance of this mechanism. Here, we developed a drug label-free, single-molecule magnetic tweezers assay that can detect a single DOX-DNA cross-link on the basis of the significant increase of the unzipping forces of DNA hairpins upon drug binding. Using this assay, we measured the DOX concentration-dependent cross-linking rates at clinically relevant concentrations of DOX. We report an â¼26-fold higher formaldehyde concentration dependence of cross-linking rates than previously reported and 0.9 ± 0.8 cross-links/103 bp at the clinically relevant concentrations of 70 nM DOX and 50 µM formaldehyde. Our results suggest a much higher cross-link formation ability than previous bulk measurements have reported and suggest that the cross-linking mechanism has promising therapeutic potential. This general method can be used to detect the formation kinetics of other DNA lesions or DNA adducts that affect DNA duplex stability.
Subject(s)
Antibiotics, Antineoplastic/analysis , Cross-Linking Reagents/chemistry , DNA/chemistry , Doxorubicin/analysis , Single Molecule Imaging , Antibiotics, Antineoplastic/pharmacology , Cell Survival/drug effects , Dose-Response Relationship, Drug , Doxorubicin/pharmacology , Formaldehyde/analysis , Formaldehyde/pharmacology , Humans , Kinetics , MCF-7 Cells , Molecular Structure , Tumor Cells, CulturedABSTRACT
A nanocomposite was prepared with reduced graphene oxide, gold nanoparticles and an electropolymerized film made from 2-amino-5-mercapto-1,3,4-thiadiazole. An electrochemical sensor for doxorubicin (DOX) was constructed by modifying a glassy carbon electrode (GCE) with the nanocomposite. The modified GCE was studied by electrochemical techniques which showed it to enable highly sensitive sensing of DOX. Response (typically measured at a typical working potential of -0.56 V vs. Ag/AgCl) is linear in the 30 pM to 30 nM and 30 nM to 30 µM DOX concentration ranges, with a limit of detection (LOD) of 9 pM (at an S/N ratio of 3). The method was applied to the determination of DOX in serum and gave recoveries that ranged between 92 and 108%. Graphical abstract A combination of materials consisting of reduced graphene oxide (rGO), gold nanoparticles (AuNPs) and an electropolymerized film of 2-amino-5-mercapto-1,3,4-thiadiazole (poly-AMT, PAMT) is described. The nanocomposite was placed on a glassy carbon elkectrode (GCE) in order to fabricate an electrochemical sensor for doxorubicin (DOX).
Subject(s)
Antibiotics, Antineoplastic/analysis , Biosensing Techniques , Doxorubicin/analysis , Electrochemical Techniques , Nanocomposites/chemistry , Gold/chemistry , Graphite/chemistry , Metal Nanoparticles/chemistry , Molecular Conformation , Oxidation-Reduction , Particle Size , Polymers/chemistry , Surface Properties , Thiadiazoles/chemistryABSTRACT
Peptide-based small molecule drug conjugates for targeted tumor therapy are currently in the focus of intensive research. Anthracyclines, like daunomycin, are commonly used anticancer drug molecules and are also often applied in peptide-drug conjugates. However, lability of the O-glycosidic bond during electrospray ionization mass spectrometric analysis hinders the analytical characterization of the constructs. "Overprotonation" can occur if daunomycin is linked to positively charged peptide carriers, like tuftsin derivatives. In these molecules, the high number of positive charges enhances the in-source fragmentation significantly, leading to complex mass spectra composed of mainly fragment ions. Therefore, we investigated different novel tuftsin-daunomycin conjugates to find an appropriate condition for mass spectrometric detection. Our results showed that shifting the charge states to lower charges helped to keep ions intact. In this way, a clear spectrum could be obtained containing intact protonated molecules only. Shifting of the protonation states to lower charges could be achieved with the use of appropriate neutral volatile buffers and with tuning the ion source parameters.
Subject(s)
Antibiotics, Antineoplastic/analysis , Daunorubicin/analysis , Glycoconjugates/analysis , Immunologic Factors/analysis , Tuftsin/analysis , Antibiotics, Antineoplastic/chemistry , Daunorubicin/chemistry , Glycoconjugates/chemistry , Humans , Immunologic Factors/chemistry , Molecular Structure , Protons , Spectrometry, Mass, Electrospray Ionization , Static Electricity , Tuftsin/chemistryABSTRACT
A metal-free catalyst is described that consists of a composite that can be prepared from mesoporous carbon spheres (MCS) and graphene oxide (GO) under mild aqueous synthetic conditions. The reduced graphene oxide (rGO) sheets tend to aggregate, but due to the insertion of MCS, the aggregation is prevented. This leads to a larger surface area and more adsorption sites for the cancer drug doxorubicin (DOX). The π-interaction between DOX and rGO is also beneficial for the adsorption of DOX. A glassy carbon electrode (GCE) was modified with the composite and used to detect low levels of DOX, typically at a peak potential near -0.45 V (vs. Ag/AgCl). The modified GCE has a wide linear response range (10 nM - 10 µM), a low limit of detection (1.5 nM; at S/N = 3), excellent selectivity, long-term storage stability and reproducibility. It was applied to the determination of DOX in spiked serum where it gave reliable results. Graphical abstract Schematic representation of the preparation of mesoporous carbon spheres/reduced graphene oxide (MCS/rGO) sample, and the CV scan of doxorubicin (DOX) on MCS/rGO based nanoprode.
Subject(s)
Antibiotics, Antineoplastic/analysis , Biosensing Techniques , Doxorubicin/analysis , Electrochemical Techniques , Nanocomposites/chemistry , Carbon/chemistry , Humans , Nanospheres/chemistry , Particle Size , Porosity , Surface PropertiesABSTRACT
Glutathione (GSH) is an important tripeptide that plays an important role in preventing damage to reactive oxygen species. An electrochemical assay was fabricated for this purpose by modification of a carbon paste electrode (CPE) with bis(1,10-phenanthroline)(1,10-phenanthroline-5,6-dione)nickel(II) hexafluorophosphate (BPPDNi) as new electro-catalyst and Pt:Co nanoparticle (Pt:CO-NPs) as highly conductive mediator. The analyses were performed at a scan rate of 10 mV/s and at a pH value of 7.4. The BPPDNi/Pt:CO-NPs/CPE showed a high sensitivity and good selectivity for electro-catalytic determination of glutathione (GSH) in nano-molar concentration range. In addition, the BPPDNi/Pt:CO-NPs/CPE was used for the determination of glutathione in the presence of doxorubicin (DOX) and tyrosine (Tyr) with three separated oxidation signals ~160 mV, ~385 mV and ~790 mV vs. Ag/AgCl/KClsat, respectively. The peak currents of the square wave voltammetric analyses were linearly dependent on glutathione, doxorubicin and tyrosine concentrations in the respective ranges of 0.001-450, 0.5-300 and 1.0-650 µM, with detection limits of 0.5 nM, 0.1 µM and 0.6 µM, respectively. Graphical abstract The first analytical sensor for simultaneous determination of glutathione, doxorubicin and tyrosine.
Subject(s)
Antibiotics, Antineoplastic/analysis , Doxorubicin/analysis , Glutathione/analysis , Tyrosine/analysis , Antibiotics, Antineoplastic/blood , Antibiotics, Antineoplastic/chemistry , Catalysis , Cobalt/chemistry , Doxorubicin/blood , Doxorubicin/chemistry , Electrochemical Techniques , Glutathione/blood , Glutathione/chemistry , Humans , Nanoparticles/chemistry , Nickel/chemistry , Platinum/chemistry , Tyrosine/blood , Tyrosine/chemistryABSTRACT
Intracellular Doppler spectroscopy is a form of low-coherence digital holography based upon Doppler detection of scattered light that measures drug response/resistance in tumor spheroids, xenografts, and clinical biopsies. Multidrug resistance (MDR) is one of the main causes of ineffective cancer treatment. One MDR mechanism is mediated by the MDR1 gene that encodes the drug efflux pump P-glycoprotein (Pgp). Overexpression of Pgp in some cancers is associated with poor chemotherapeutic response. This paper uses intracellular Doppler spectroscopy to detect Pgp-mediated changes to drug response in 3D tissues grown from an ovarian cancer cell line (SKOV3). The SKOV3 cell line was incrementally exposed to cisplatin to create a cell line with increased Pgp expression (SKOV3cis). Subsequently, MDR1 in a subset of these cells was silenced in SKOV3cis using shRNA to create a doxycycline inducible, Pgp-silenced cell line (SKOV3cis-sh). A specific Pgp inhibitor, zosuquidar, was used to study the effects of Pgp inhibition on the Doppler spectra. Increased drug sensitivity was observed with Pgp silencing or inhibition as determined by drug IC50s of paclitaxel-response of silenced Pgp and doxorubicin-response of inhibited Pgp, respectively. These results indicate that intracellular Doppler spectroscopy can detect changes in drug response due to silencing or inhibition of a single protein associated with drug resistance with important consequences for personalized medicine.
Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/antagonists & inhibitors , Antibiotics, Antineoplastic/pharmacology , Dibenzocycloheptenes/pharmacology , Doxorubicin/pharmacology , Laser-Doppler Flowmetry , Ovarian Neoplasms/drug therapy , Quinolines/pharmacology , Spheroids, Cellular/drug effects , ATP Binding Cassette Transporter, Subfamily B/antagonists & inhibitors , ATP Binding Cassette Transporter, Subfamily B/genetics , ATP Binding Cassette Transporter, Subfamily B, Member 1/analysis , ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , Antibiotics, Antineoplastic/analysis , Cell Proliferation/drug effects , Cell Survival/drug effects , Dibenzocycloheptenes/chemistry , Doxorubicin/analysis , Drug Screening Assays, Antitumor , Female , Gene Silencing/drug effects , Humans , Ovarian Neoplasms/diagnostic imaging , Quinolines/chemistry , Tumor Cells, CulturedABSTRACT
Anticancer drugs are prescribed and administrated to an increasing number of patients on a daily basis. As a consequence, a number of concerns have been raised about the patient health and safety in the case that the drugs administered are not at the required concentration or even worse not the correct ones. Quality control of therapeutic solutions has therefore been extensively implemented in hospital environments, in order to avoid any failure in the intense workflow faced by administering pharmacists. In the present study, infrared (IR) and Raman spectroscopy have been employed for the analysis of 3 commercially available therapeutic solutions TEVA®, MYLAN®, CERUBIDINE®, respectively containing doxorubicin, epirubicin and daunorubicin. They perfectly illustrate the analytical difficulties encountered, as these 3 chemotherapeutic drugs are isomers, hardly distinguishable with conventional approaches such as UV/VIS spectrometry. Any analytical failure to identify these molecules can lead to delays in patient treatment. While Partial Least Squares Regression analysis demonstrates that both Raman and IR can deliver satisfactory quantitative analysis in the clinical range, with respective Root Mean Square Error of Cross Validation (RMSECV) between 0.0127 - 0.0220â¯g·L-1 and 0.0573 - 0.0759â¯g·L-1, the identification rate between the 2 techniques differs substantially. Indeed, Principal Component Analysis - Factorial Discriminant Analysis (PCA-FDA) highlights that, depending on the data preprocessing applied to Raman spectra, the discrimination between the 3 drugs is decreased, with in some cases specificity and sensitivity below 50%. However, IR analysis displays encouraging results with an overall specificity and sensitivity between 99 and 100%, suggesting that reliable validation of the therapeutic solution for administration to patients can be achieved. IR and Raman spectroscopy could assist and support quality control of chemotherapeutic solutions prepared in personalised concentrations for each patient. The effective and reliable characterisation of therapeutic solutions could have a lot to offer to improve current practices in a near future.
Subject(s)
Antibiotics, Antineoplastic/analysis , Daunorubicin/analysis , Doxorubicin/analysis , Epirubicin/analysis , Spectrophotometry, Infrared/methods , Spectrum Analysis, Raman/methods , Discriminant Analysis , Principal Component Analysis , SolutionsABSTRACT
MHC-I peptides are intracellular-cleaved peptides, usually 8-11 amino acids in length, which are presented on the cell surface and facilitate CD8+ T cell responses. Despite the appreciation of CD8+ T-cell antitumor immune responses toward improvement in patient outcomes, the MHC-I peptide ligands that facilitate the response are poorly described. Along these same lines, although many therapies have been recognized for their ability to reinvigorate antitumor CD8+ T-cell responses, whether these therapies alter the MHC-I peptide repertoire has not been fully assessed due to the lack of quantitative strategies. We develop a multiplexing platform for screening therapy-induced MHC-I ligands by employing tandem mass tags (TMTs). We applied this approach to measuring responses to doxorubicin, which is known to promote antitumor CD8+ T-cell responses during its therapeutic administration in cancer patients. Using both in vitro and in vivo systems, we show successful relative quantitation of MHC-I ligands using TMT-based multiplexing and demonstrate that doxorubicin induces MHC-I peptide ligands that are largely derived from mitotic progression and cell-cycle proteins. This high-throughput MHC-I ligand discovery approach may enable further explorations to understand how small molecules and other therapies alter MHC-I ligand presentation that may be harnessed for CD8+ T-cell-based immunotherapies.
Subject(s)
Antibiotics, Antineoplastic/analysis , Colonic Neoplasms/therapy , Doxorubicin/analysis , Histocompatibility Antigens Class I/analysis , Lymphoma/therapy , Animals , Antibiotics, Antineoplastic/pharmacology , CD8-Positive T-Lymphocytes/drug effects , CD8-Positive T-Lymphocytes/immunology , Colonic Neoplasms/immunology , Doxorubicin/pharmacology , Drug Discovery , HCT116 Cells , Histocompatibility Antigens Class I/immunology , Humans , Immunotherapy , Ligands , Lymphoma/immunology , Mass Spectrometry , Mice , Mice, Inbred C57BL , Tumor Cells, CulturedABSTRACT
A simple, straightforward analytical method based on liquid chromatography has been optimized to quantify total, internal, and external ions in drug-loaded liposomal products. The quantification of ammonium and sulfate ions in Doxil is detailed; although, the methodology has been extrapolated to quantitate a variety of ions, including calcium, acetate, and others in several different liposomal formulations. Total ion concentrations were measured after disruption of the liposome via lyophilization, to liberate all components. External ion concentrations were made following membrane centrifugation, without disruption of the liposome structure, where the permeate fraction was analyzed for external ion quantities. The internal ion fraction was derived from mass balance of the total and external ion measurements. High performance liquid chromatography (HPLC), equipped with different separation columns, and coupled to a charged aerosol detector, was employed for all ion quantifications. The analytical measurements were confirmed using simple stoichiometry based on the drug crystallization of doxorubicin within the liposome interior. The method presented herein is quick, highly accurate, and has significantly improved lower limits of detection and quantification over other traditional methods. As more follow-on versions of Doxil are being developed, this facile approach to ion quantitation can be used to help establish compositional similarity to the reference listed drug.
Subject(s)
Antibiotics, Antineoplastic/analysis , Chromatography, High Pressure Liquid/methods , Doxorubicin/analogs & derivatives , Antibiotics, Antineoplastic/chemistry , Crystallization , Doxorubicin/analysis , Doxorubicin/chemistry , Drug Delivery Systems , Freeze Drying , Ions , Limit of Detection , Liposomes , Polyethylene Glycols/analysis , Polyethylene Glycols/chemistry , Reproducibility of ResultsABSTRACT
Doxorubicin (DOX) is a highly effective anthracycline antibiotic. Unfortunately, the clinical use of DOX is limited by the risk of deleterious effects to cardiac and respiratory (i.e. diaphragm) muscle, resulting from mitochondrial reactive oxygen species (ROS) production. In this regard, exercise is demonstrated to protect against DOX-induced myotoxicity and prevent mitochondrial dysfunction. However, the protective mechanisms are currently unclear. We hypothesized that exercise may induce protection by increasing the expression of mitochondria-specific ATP-binding cassette (ABC) transporters and reducing mitochondrial DOX accumulation. Our results confirm this finding and demonstrate that two weeks of exercise preconditioning is sufficient to prevent cardiorespiratory dysfunction.
Subject(s)
Antibiotics, Antineoplastic/analysis , Diaphragm/chemistry , Doxorubicin/analysis , Mitochondria/chemistry , Myocardium/chemistry , Physical Conditioning, Animal , Animals , Antibiotics, Antineoplastic/pharmacokinetics , Doxorubicin/pharmacokinetics , Female , Rats, Sprague-DawleyABSTRACT
A simple, rapid, reliable and sensitive method based on liquid chromatography with fluorescence detection (LC-FL) for the quantification of doxorubicin (DOX) in human plasma and urine samples was developed. The assay was carried out after the solid-phase extraction procedure (SPE) with hydrophilic-lipophilic balance (HLB) cartridges, and with daunorubicin hydrochloride (DAU) used as the internal standard. Chromatographic separation was performed on a Discovery HS C18 column in isocratic elution mode, and the detection of the analytes set at excitation and emission wavelengths of 487 and 555â¯nm, respectively. The developed LC-FL method has been validated for accuracy, precision, selectivity, linearity, recovery and stability. The limits of detection and quantification for DOX were 0.5 and 1â¯ng/mL in both biological fluids, respectively. Linearity was confirmed in the range of 1-1000â¯ng/mL and 0.001-25⯵g/mL in plasma and urine samples, respectively, with a correlation coefficient greater than 0.9994. The proposed LC-FL method is selective, precise and accurate, and has been successfully applied for drug monitoring in pediatric cancer patients treated with DOX as a component of OEPA (Oncovin (Vincristine)-Etoposide-Prednisone-Adriamycin) and IOA (Ifosfamide-Oncovin-Adriamycin) chemotherapeutic schemes. Moreover, real exposure of hospital personnel to the anthracycline drugs in plasma and urine was evaluated in clinical practice.
Subject(s)
Antibiotics, Antineoplastic/analysis , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Doxorubicin/analysis , Drug Monitoring/methods , Neoplasms/drug therapy , Adolescent , Antibiotics, Antineoplastic/therapeutic use , Antibiotics, Antineoplastic/toxicity , Chromatography, High Pressure Liquid/instrumentation , Chromatography, High Pressure Liquid/methods , Daunorubicin/analysis , Doxorubicin/pharmacokinetics , Doxorubicin/therapeutic use , Doxorubicin/toxicity , Etoposide/therapeutic use , Fluorescence , Humans , Limit of Detection , Male , Neoplasms/blood , Neoplasms/urine , Occupational Exposure/adverse effects , Personnel, Hospital , Prednisone/therapeutic use , Reproducibility of Results , Solid Phase Extraction , Vincristine/therapeutic useABSTRACT
Fluorescent copper nanoparticles (CuNPs) have received great attention due to their distinct characteristics of facile synthesis, tunable fluorescence emission, high photostability, and biological compatibility, and they have been widely used for chemical and biological analyses. Bleomycins (BLMs) are widely used antitumor agents for the clinical treatment of various cancers. Here, we develop a sensitive and label-free fluorescence method for the quantitative detection of BLM on the basis of BLM-initiated enzymatic polymerization-mediated synthesis of fluorescent CuNPs. We design two hairpin DNAs: one (Hp1) for the recognition of BLM and the other (Hp2) for signal amplification. In the presence of BLM, it may recognize and cleave the 5'-GC-3' site of the Hp1 stem, releasing the 8-17 DNAzyme fragment. The resultant 8-17 DNAzyme fragments may bind with the loop of Hp2 to form a partial double-stranded DNA (dsDNA) duplex, initiating the cyclic cleavage of Hp2 in the presence of Zn2+-dependent DNAzymes and generating numerous new DNA fragments with the free 3'-OH terminal, which can induce the formation of a poly(thymine) (poly-T) sequence with the assistance of terminal deoxynucleotidyl transferase (TdTase). Subsequently, the ploy-T sequence may function as the template for the synthesis of CuNPs with strong fluorescence emission. This method shows good selectivity and high sensitivity with a detection limit as low as 8.1 × 10-16 M, and it exhibits good performance in serum samples. Moreover, this method has distinct advantages of simplicity and low cost, holding great potential in clinical diagnosis and biomedical research.
Subject(s)
Bleomycin/analysis , Copper , DNA, Catalytic/chemistry , Metal Nanoparticles , Poly T/chemistry , Antibiotics, Antineoplastic/analysis , DNA Nucleotidylexotransferase/chemistry , Spectrometry, FluorescenceABSTRACT
In this work, a simple room-temperature phosphorescence (RTP) method was first proposed to detect adriamycin, by using a functional material of Mn doped ZnS quantum dots (Mn-ZnS QDs) composited with poly(diallyldimethylammonium chloride) (PDDA). After PDDA was associated to the Mn-ZnS QDs, the RTP intensity was significantly enhanced. As a result, Mn-ZnS QDs/PDDA nanoassemblies greatly improve the adriamycin detection ability of QDs and provide an important method for developing more effective and sensitive adriamycin detection sensor. The method based upon RTP has a linear range from 0.5 to 64.0 µM with a detection limit (3σ/s) for adriamycin of 0.45 µM. The developed method was further successfully applied to detect adriamycin in human serum samples (diluted 50 times) with satisfactory results, and the recovery ranged from 98.3 to 101.3.