BACKGROUND: Microsampling of biological fluids followed by innovative sample pre-treatment reflects trends in bioanalytical chemistry. Volumetric absorptive microsampling (VAMS) enables exact whole blood volume collection and reduces the impact of hematocrit on the assay. In animal studies, it complies with the 3R principles (refine, reduce, replace). It allows for a gentle bleeding technique and a reduction in the number of laboratory animals by enabling ethically acceptable repeated blood collection from a single animal. Treating VAMS tips with electromembrane extraction (EME) in 96-well format offers a smart combination of non-invasive, low-volume blood collection with effective, environmentally friendly sample clean-up. RESULTS: This study introduces the first application of EME in 96-well format for direct isolation of analytes from 10 μL of whole blood collected onto a VAMS device. Doxorubicin, a clinically used anticancer drug also utilized in cancer/cardio-oncology research involving rodents, where microsampling offers important advantages, and its metabolite doxorubicinol, were selected as relevant analytes. The optimized EME yielded reproducible recoveries for both analytes regardless of hematocrit levels, different anticoagulants, or free multivalent ions in the sample. Compared to conventional VAMS tips treatment, EME reduced matrix effects, increased throughput, and an environmental friendliness of the extraction. The EME followed by the UHPLC-MS/MS assay was validated for both analytes in whole blood absorbed onto VAMS tips. The same protocol was implemented to treat plasma to determine the blood-to-plasma ratio of the analytes in the same experiments. The practical utility was demonstrated by analyzing real samples collected from the doxorubicin-treated nude mice. SIGNIFICANCE: The study offers a novel assay combining whole blood microsampling and sample clean-up in microextraction scale for preclinical pharmacokinetic studies with doxorubicin in rodents and for pharmacokinetic/pharmacodynamic modeling. This advancement in bioanalytical chemistry promotes scalable environmentally friendly procedures compatible with the 3R ethical principles in animal studies. Moreover, the concept of direct VAMS tips treatment with EME may also be easily translatable to clinical settings.

• Klíčová slova
• Anthracyclines, Doxorubicin, Doxorubicinol, Electromembrane extraction, Microextraction, Volumetric absorptive microsampling,
• MeSH
• doxorubicin * krev   analogy a deriváty   chemie   MeSH
• elektrochemické techniky metody   MeSH
• membrány umělé MeSH
• odběr vzorku krve metody   MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• doxorubicin * MeSH
• membrány umělé MeSH

INTRODUCTION: Nanoparticle-based drug delivery systems can overcome the dose-limited toxicity of cytostatics. Pegylated doxorubicin-containing liposomes (PLD) are able to reduce cardiotoxicity. PLD quickly (in 2 days) attains therapeutic concentration in tumorous tissue (kinetic targeting), while its distribution in normal tissue, which is a cause of mucocutaneous toxicity (MCT), is delayed. We examined PLD extracorporeal removal effectivity, using plasma filtration (PF) to determine whether the drug could be withheld prior to its organ distribution responsible for MCT toxicity. METHODS: Nine patients suffering from platinum-resistant ovarian cancer were treated with a infusion of 50 mg/m2 of PLD/cycle - for four cycles q4w. Over 44 (46)-47 (49) hours postinfusion, the patients (14 cycles in total) underwent PF using the cascade method. Doxorubicin blood concentration was monitored by the HPLC method during 116 h. Individual pharmacokinetic parameters of doxorubicin were estimated. RESULTS: Over 44 (46)-47 (49) hours postinfusion, a single one-volume plasma filtration removed 35 (22-45) % of the remaining doxorubicin amount in the body. Symptoms of MCT - PPE-like syndrome (grade 3) appeared in one patient. Only one adverse reaction (1/14-7%) - short-term malaise and nausea - was reported as being related to PF. CONCLUSION: PF does remove a clinically important amount of doxorubicin in a kinetic targeting approach, which can be a useful tool for the increased efficacy and tolerability of therapy with PLD. There were no serious signs of drug toxicity and/or PF-related adverse events.

• Klíčová slova
• Kinetic targeting, Ovarian cancer, Pegylated doxorubicin, Pharmacokinetic, Plasma filtration,
• MeSH
• chemorezistence MeSH
• dospělí MeSH
• doxorubicin aplikace a dávkování   škodlivé účinky   analogy a deriváty   krev   farmakologie   MeSH
• intravenózní infuze MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory vaječníků krev   diagnóza   farmakoterapie   MeSH
• polyethylenglykoly aplikace a dávkování   škodlivé účinky   farmakologie   MeSH
• příprava léků MeSH
• protinádorová antibiotika aplikace a dávkování   škodlivé účinky   krev   farmakokinetika   MeSH
• senioři MeSH
• tkáňová distribuce MeSH
• výměna plazmy metody   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• doxorubicin MeSH
• liposomal doxorubicin MeSH Prohlížeč
• polyethylenglykoly MeSH
• protinádorová antibiotika MeSH

BACKGROUND: One approach to improve effect of chemotherapy is combination of classical cytostatic drugs with natural compounds, e. g. sesquiterpenes. In our previous study, sesquiterpenes β-caryophyllene oxide (CAO) and trans-nerolidol (NER) improved the anti-proliferative effect of doxorubicin (DOX) in intestinal cancer cell lines. PURPOSE: The present study was designed to evaluate the effect of CAO and NER on DOX efficacy, focusing on cell proliferation, migration, apoptosis and DOX accumulation in breast cancer cells MDA-MB-231 and MCF7 in vitro and in mice bearing solid Ehrlich tumors (EST) in vivo. METHODS: The impact of cytotoxic effect was assessed by the neutral red uptake test. The ability to migrate was tested using real-time measurement in x-CELLigence system. Expressions of molecules were examined using western blot analysis. The accumulation of DOX inside the cells using time lapse microscopy was observed. The mice with inoculated EST cells were treated repeatedly with DOX and DOX+CAO or DOX+NER and the growth of tumors were monitored. DOX concentrations in plasma and tumor were assayed using HPLC. RESULTS: In MDA-MB-231, combination of DOX with CAO enhanced anti-proliferative effect and acted strongly synergistic. NER increased accumulation of DOX inside the cells; moreover combination DOX with NER suppressed migration ability in vitro. In vivo, apoptosis was activated especially in group treated with DOX and CAO. However, none of tested sesquiterpenes was able to improve DOX accumulation in tumors and DOX-mediated inhibition of tumor growth. CONCLUSION: In conclusion, sesquiterpenes CAO and NER increased the efficacy of DOX in breast cancer cells in vitro, but did not improve its effect in vivo, in Ehrlich solid tumor bearing mice.

• Klíčová slova
• Anti-apoptotic effect, Breast cancer cells, Doxorubicin, Trans-nerolidol, Tumor bearing mice, β-Caryophyllene oxide,
• MeSH
• doxorubicin krev   terapeutické užití   MeSH
• inhibiční koncentrace 50 MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory mléčné žlázy u zvířat farmakoterapie   patologie   MeSH
• nádory prsu krev   farmakoterapie   patologie   MeSH
• pohyb buněk účinky léků   MeSH
• polycyklické seskviterpeny MeSH
• proliferace buněk účinky léků   MeSH
• seskviterpeny chemie   farmakologie   terapeutické užití   MeSH
• výsledek terapie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• caryophyllene oxide MeSH Prohlížeč
• doxorubicin MeSH
• nerolidol MeSH Prohlížeč
• polycyklické seskviterpeny MeSH
• seskviterpeny MeSH

PURPOSE: To examine the removal of pegylated liposomal doxorubicin (PLD) during plasmafiltration (PF) and determine whether the drug could be withheld prior to its organ distribution responsible for mucocutaneous toxicity. METHODS: Six patients suffering from platinum-resistant ovarian cancer were treated with a 1-h IV infusion 50 mg/m(2) of PLD/cycle-for three cycles q4w. Over 44 (46)-47(49) h postinfusion, five patients (14 cycles in total) underwent PF using a cascade PF method consisted of plasma separation by centrifugation and plasma treatment using filtration based one volume of plasma treatment, i.e., 3.18 L (±0.6 L) and plasma flow 1.0 L/h (0.91-1.48 L/h). Doxorubicin concentration in blood was monitored by a high-performance liquid chromatography method for 116 h postinfusion. Pharmacokinetic parameters determined from plasma concentration included volume of distribution, total body clearance, half-life of elimination, and area under the plasma concentration versus time. The amount of doxorubicin in the body eliminated by the patient and via extracorporeal treatment was evaluated. Toxicity was tested using CTCAE v4.0. RESULTS: The efficacy of PF and early responses to PLD/PF combination strategy were as follows: over 44(46) h postinfusion considered necessary for target distribution of PLD to tumor, patients eliminated 46 % (35-56 %) of the dose administered. Over 44(46)-47(49) h postinfusion, a single one-volume plasma filtration removed 40 % (22-45 %) (Mi5) of the remaining doxorubicin amount in the body. Total fraction eliminated attained 81 % (75-86 %). The most common treatment-related adverse events (grade 1-2) such as nausea (4/14 cycles-28 %) and vomiting (3/14 cycles-21 %) appeared during 44 h postinfusion. Hematological toxicity-anemia (5/14 cycles-35 %) was reported after cycle II termination. Symptoms of PPE-like syndrome (grade 1-2) appeared in one patient concomitantly with thrombophlebitis and malignant effusion. In this study, only one adverse reaction (1/14-7 %) as short-term malaise and nausea was reported by the investigator as probably related to PF. CONCLUSION: A single one-volume PF does remove a clinically important amount of doxorubicin in a kinetic targeting approach. There were no serious signs of drug toxicity and/or PF-related adverse events. Kinetically guided therapy with pegylated liposomal doxorubicin combined with PF may be a useful tool to the higher efficacy and tolerability of therapy with PLD.

• Klíčová slova
• Kinetic targeting, Pegylated liposomal doxorubicin, Plasmafiltration,
• MeSH
• dospělí MeSH
• doxorubicin aplikace a dávkování   škodlivé účinky   analogy a deriváty   krev   farmakokinetika   MeSH
• hemofiltrace škodlivé účinky   metody   MeSH
• kritické orgány MeSH
• lidé středního věku MeSH
• lidé MeSH
• metabolická clearance MeSH
• nádory vaječníků * farmakoterapie   patologie   MeSH
• nádory vejcovodů * farmakoterapie   patologie   MeSH
• nežádoucí účinky léčiv etiologie   prevence a kontrola   MeSH
• plocha pod křivkou MeSH
• poločas MeSH
• polyethylenglykoly aplikace a dávkování   škodlivé účinky   farmakokinetika   MeSH
• progrese nemoci MeSH
• protinádorová antibiotika aplikace a dávkování   škodlivé účinky   krev   farmakokinetika   MeSH
• senioři MeSH
• staging nádorů MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• doxorubicin MeSH
• liposomal doxorubicin MeSH Prohlížeč
• polyethylenglykoly MeSH
• protinádorová antibiotika MeSH

Ovarian cancer is the fifth most common malignancy in the world's female population and with the highest lethality index among gynecological tumors. The prognosis of metastatic disease is usually poor, especially in platinum-resistant cases. There are several options for the treatment of metastatic disease resistant to platinum derivates (e.g. paclitaxel, topotecan and pegylated liposomal doxorubicin), all of which are considered equipotent. Pegylated liposomal doxorubicin (PLD) is a liposomal form of the anthracycline antibiotic doxorubicin. It is characterized by more convenient pharmacokinetics and a different toxicity profile. Cardiotoxicity, the major adverse effect of conventional doxorubicin, is reduced in PLD as well as hematotoxicity, alopecia, nausea and vomiting. Skin toxicity and mucositis, however, emerge as serious issues since they represent dose and schedule-limiting toxicities. The pharmacokinetics of PLD (prolonged biological half-life and preferential distribution into tumor tissue) provide new possibilities to address these toxicity issues. The extracorporeal elimination of circulating liposomes after PLD saturation in the tumor tissue represents a novel and potent strategy to diminish drug toxicity. This article intends to review PLD characteristics and the importance of extracorporeal elimination to enhance treatment tolerance and benefits.

• Klíčová slova
• Palmar-plantar erythrodysesthesia, Pegylated liposomal doxorubicin, Rheopheresis, Toxicity,
• MeSH
• chemorezistence MeSH
• cytostatické látky MeSH
• doxorubicin analogy a deriváty   krev   terapeutické užití   MeSH
• lidé MeSH
• mimotělní oběh * MeSH
• nádory vaječníků krev   farmakoterapie   MeSH
• polyethylenglykoly terapeutické užití   MeSH
• protinádorová antibiotika krev   terapeutické užití   MeSH
• sloučeniny platiny terapeutické užití   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Názvy látek
• cytostatické látky MeSH
• doxorubicin MeSH
• liposomal doxorubicin MeSH Prohlížeč
• polyethylenglykoly MeSH
• protinádorová antibiotika MeSH
• sloučeniny platiny MeSH

Alcoholic liver disease may be frequently complicated by mesangial proliferation with the deposition of IgA in glomeruli and glomerulosclerosis, but these glomerular lesions are usually mild and without greater impact on renal function. To evaluate the putative role of ethanol in glomerular pathology we studied the influence of chronic ethanol administration on the development of experimental adriamycin nephropathy in rats. Nephrotic syndrome was induced by a single i.v. dose of adriamycin (5 mg/kg body wt) both in rats given ethanol at a dose of 4 g/day for 3 months and control rats given standard chow. Further controls on both diets without adriamycin administration were also studied. Blood and urine were examined before and 3 and 6 weeks after adriamycin administration. All rats were killed and examined histologically 6 weeks after adriamycin administration. Ethanol fed nephrotic rats were more catabolic than control nephrotic rats (with higher free fatty acids, lower glycaemia, higher urea with similar creatinine) and had lower proteinuria (0.55 +/- 0.34 versus 5.79 +/- 3.15 g of protein/nmol of creatinine, P < 0.05), higher albuminaemia (5.41 +/- 2.62 versus 1.92 +/- 1.94 g/l, P < 0.01), lower plasma cholesterol (6.54 +/- 2.6 versus 10.57 +/- 2.92 mmol/l, P < 0.01) and triglycerides. The development of nephrotic syndrome and renal morphological changes after adriamycin administration in rats seemed to be ameliorated, or at least delayed by chronic ethanol feeding with much milder and focal glomerulosclerosis as compared with more severe and diffuse glomerulosclerosis in control nephrotic animals. The mechanism of this effect of chronic ethanol feeding remains to be elucidated.(ABSTRACT TRUNCATED AT 250 WORDS)

• MeSH
• aspartátaminotransferasy metabolismus   MeSH
• cholesterol krev   MeSH
• doxorubicin škodlivé účinky   krev   moč   MeSH
• ethanol aplikace a dávkování   farmakologie   MeSH
• glomerulus MeSH
• HDL-cholesterol MeSH
• hyperglykemie MeSH
• kreatin krev   moč   MeSH
• krysa rodu Rattus MeSH
• ledviny cytologie   účinky léků   patofyziologie   MeSH
• nefrotický syndrom chemicky indukované   patofyziologie   MeSH
• potkani Wistar * MeSH
• vztah mezi dávkou a účinkem léčiva * MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• aspartátaminotransferasy MeSH
• cholesterol MeSH
• doxorubicin MeSH
• ethanol MeSH
• HDL-cholesterol MeSH
• kreatin MeSH

A method for the determination of doxorubicin and daunorubicin in plasma is described. The plasma is injected directly into a loop column and then washed with water. After switching the injection valve, the sample is separated on a phenyl column using detection at 254 nm. The detection limit is 10 ng/mL, the coefficient of variation is 7% for 100 ng/mL of doxorubicin and 4% for 200 ng/mL of daunorubicin.

• MeSH
• daunomycin krev   MeSH
• doxorubicin krev   MeSH
• lidé MeSH
• spektrofotometrie ultrafialová MeSH
• vysokoúčinná kapalinová chromatografie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• daunomycin MeSH
• doxorubicin MeSH