Methotrexate Solution Research Articles

Influence of particle size on transport of methotrexate across blood brain barrier by polysorbate 80-coated polybutylcyanoacrylate nanoparticles

Transports of methotrexate-loaded polybutylcyanoacrylate nanoparticles with different sizes across blood brain barrier were investigated in this experiment. The drug-loaded nanoparticles were prepared by emulsion polymerization method. After coating with polysorbate 80, nanoparticles with the size 70, 170, 220, 345 nm were, respectively, i.v. injected into rats at the dose of 3.2 mg/kg. Uncoated nanoparticles and methotrexate solution were also i.v. injected at the same dosage as controls. 0.5, 1, 1.5, 2, 3, 4 h after injection, cerebrospinal fluids and brain tissues were collected for tests. Drug level in all biological samples was determined by HPLC. It was found out that nanoparticles overcoated by polysorbate 80 could significantly improve the drug level in both brain tissues and cerebrospinal fluids compared with uncoated ones and simple solution. Seventy-nanometer nanoparticles could deliver more drugs into brain while no significant difference was observed among the other three size ranges. In conclusion, polysorbate 80-coated polybutylcyanoacrylate nanoparticles could be used to overcome blood brain barrier especially those whose diameter was below 100 nm.

Pharmacokinetic Study of Methotrexate Following Intra-Articular Injection of Methotrexate Loaded Poly(L-Lactic Acid) Microspheres in Rabbits

The pharmacokinetics and tissue distribution of methotrexate (MTX) were investigated following intra-articular injection of either MTX solution or controlled release MTX loaded microspheres in healthy rabbit joints. MTX solution or MTX loaded microspheres (size 30-100 mum) (10 mg MTX) was injected into the right knee joint cavity of rabbits. Blood samples were taken at predetermined times from the jugular vein. Urine samples were also collected over time periods up to 24 h. The major organs and synovial tissues were removed for analysis 6 and 24 h post-injection (n = 4). MTX and 7-OH-MTX concentrations in the plasma and major organs were determined by HPLC. The MTX plasma area under the concentration-time curve (AUC) for rabbits injected with MTX solution was seven fold higher than that of the rabbits injected with MTX microspheres, while t(1/2) and mean residence time (MRT) were not significantly different between two treatment groups. Four fold more MTX was excreted in the urine from rabbits injected with MTX solution compared to those injected with MTX loaded microspheres 24 h following intra-articular injection. The concentration of MTX in the synovial tissues following intra-articular injection was significantly higher in the rabbits injected with microspheres than in the rabbits injected with MTX solution. MTX solution was rapidly cleared from the joint cavity while MTX encapsulated microspheres retained MTX in the joint cavity.

Study of stability of methotrexate in acidic solution Spectrofluorimetric determination of methotrexate in pharmaceutical preparations through acid-catalyzed degradation reaction

Study of the degradation reaction of methotrexate (MTX) in acidic solution was carried out. Optimization of the experimental parameters of MTX acid hydrolysis was investigated. Spectrofluorimetric method for determination of MTX through measurement of its acid-degradation product, 4-amino-4-deoxy-10-methylpteroic acid (AMP), was developed. Stability of the standard solution of MTX prepared in sulfuric acid was discussed in the view of accelerated stability analysis. Two other comparative spectroflourimetric methods based on measuring the fluorescence intensities from either a condensation reaction with acetylacetone–formaldehyde (Hantzsch reaction) or a reaction with fluorescamine were also described. Beer's law validation, accuracy, precision, limits of detection, limits of quantification, and other aspects of analytical merit are presented in the text. The proposed methods were successfully applied for the analysis of MTX in pure drug and tablets dosage form. The sensitivity of the developed methods was favorable, so it was possible to be adopted for determination of MTX in plasma samples for routine use in high-dose MTX therapy.

Improved Efficacy of a Microencapsulated Macrophage Colony Stimulating Factor and Methotrexate in Melanoma

This study examines the effects of a combination therapy of both methotrexate (MTX) and albumin microspheres containing recombinant human macrophage colony-stimulating factor (rhM-CSF) in melanoma tumors. Melanoma tumors were induced in C57BL/6 male mice with subcutaneous injection of B-16 tumor cells. Therapy started once the tumor size reached 0.5 cm in diameter. Mice were divided into several groups, and dosing was carried out daily until death. Group I received MTX solution (2 mg/kg or 15 mg/kg), group II received rhM-CSF solution (100 μg), group III received albumin rhM-CSF microspheres (100 μg), and groups V–XV received different combinations of both agents daily. The weight, tumor size, and survival time (in days) were recorded. From the results, the control (no rhM-CSF administered) group survived for 11.8 ± 1.92 days, and the group that received MTX solution survived for 19.4 ± 5.03 days. However, the group that received both the MTX solution (15 mg/kg) and albumin rhM-CSF microspheres (100 μg/kg) demonstrated a significant increase (p <. 05) in the survival time (30.4 ± 3.27 days). The concentrations of cytokines (tumor necrosis factor alpha [TNF-α] and interleukin-1 beta [IL-1β]) in the different treatment groups were monitored to determine the effect of rhM-CSF on the immune system. The TNF-α concentration was significantly higher in the group that received the combination therapy (204 ± 54.6 pg/ml) versus the control group (31.5 ± 7.02 pg/ml). The IL-1β concentration was significantly higher (p <. 05) in the rhM-CSF microsphere (100 μg/kg) treated group (62 ± 17.2 pg/ml) versus the rhM-CSF solution (29.1 ± 8.7 pg/cc).

Development of porous apatite ceramic for local delivery of chemotherapeutic agents.

An experimental study was conducted on a drug delivery system (DDS), using porous apatite ceramics (PAC): hydroxyapatite block (HAb) [Ca10(PO4)6(OH)2] having a porosity of 35-48% and pore size range of 50-300 microm, and beta-tricalcium phosphate block (TCP) [Ca3(PO4)2] having a porosity of 75-80% and pore size range of 100-400 microm, for sustained release of a chemotherapeutic agent. Methotrexate (MTX) was loaded in the pores of PAC blocks by centrifuging the blocks in MTX solution. Impregnation of MTX in PAC blocks (1 cm3) was confirmed by a magnetic resonance imaging (MRI) study using Gadolinium-DTPA enhancement. The MRI showed high signal intensity in the PAC, which was confirmed by dye loading into the pores. To estimate the MTX-releasing capability of the PAC, the blocks were stored in 3 mL of phosphate-buffered saline (PBS) at 37 degrees C and the PBS was replaced every 48 h. The amount of MTX released was assayed by high-performance liquid chromatography. This study showed that MTX-impregnated PAC (0.63-2.25 mg/block) released the drug in a steady manner and maintained its concentration (0.1-1.0 microg/mL) up to 12 days. This concentration is high enough to be effective against tumor cells. Chemotherapeutic agent-impregnated PAC, prepared by simple centrifugation, could be a valuable form of local chemotherapy when used as a strut graft to repair bone defects. This new DDS material could also be used as an adjuvant to extended curettage and provide a means to reduce the recurrence of tumors without risk of systemic toxicity.

Development of porous apatite ceramic for local delivery of chemotherapeutic agents

An experimental study was conducted on a drug delivery system (DDS), using porous apatite ceramics (PAC): hydroxyapatite block (HAb) [Ca10(PO4)6(OH)2] having a porosity of 35–48% and pore size range of 50–300 μm, and β-tricalcium phosphate block (TCP) [Ca3(PO4)2] having a porosity of 75–80% and pore size range of 100–400 μm, for sustained release of a chemotherapeutic agent. Methotrexate (MTX) was loaded in the pores of PAC blocks by centrifuging the blocks in MTX solution. Impregnation of MTX in PAC blocks (1 cm3) was confirmed by a magnetic resonance imaging (MRI) study using Gadolinium-DTPA enhancement. The MRI showed high signal intensity in the PAC, which was confirmed by dye loading into the pores. To estimate the MTX-releasing capability of the PAC, the blocks were stored in 3 mL of phosphate-buffered saline (PBS) at 37°C and the PBS was replaced every 48 h. The amount of MTX released was assayed by high-performance liquid chromatography. This study showed that MTX-impregnated PAC (0.63–2.25 mg/block) released the drug in a steady manner and maintained its concentration (0.1–1.0 μg/mL) up to 12 days. This concentration is high enough to be effective against tumor cells. Chemotherapeutic agent–impregnated PAC, prepared by simple centrifugation, could be a valuable form of local chemotherapy when used as a strut graft to repair bone defects. This new DDS material could also be used as an adjuvant to extended curettage and provide a means to reduce the recurrence of tumors without risk of systemic toxicity. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 39, 536–538, 1998.

Stability and compatibility of 2.5 mg/ml methotrexate solution in plastic syringes over 7 days

The physico-chemical stability and the compatibility between a 2.5-mg/ml methotrexate solution and plastic syringes were studied over a 7 day period at + 4 and 25°C. During this study, pH was determined and MTX degradation products were assayed by HPLC. The compatibility study consisted of the titration of reducing substances, TLC of extracted additives, assay of extracted ions by AAS, resistivity measurement, dynamometric test and UV, visible and IR absorption spectrophotometry. Stability and compatibility studies showed that 2.5 mg/ml methotrexate parenteral solution remained chemically stable in a plastic syringe over a period of 7 days under the tested conditions.

Effect of Niosome Encapsulation of Methotrexate, Macrophage Activation on Tissue Distribution of Methotrexate and Tumor Size

AbstractMethotrexate was encapsulated in “niosomes” of sorbitan monostearate (Span 60) by the reverse phase evaporation technique. After i.v. infusion into mice methotrexate concentration was higher in all tissues except kidney with niosomal methotrexate than with methotrexate solution. Niosome encapsulation and macrophage activation enhanced the delivery of methotrexate to the tumor and increased its antitumor activity.

Intratunical versus parenteral administration of methotrexate.

Methotrexate (MXT) distribution in serum, testicle and epididymis after administration into the tunica vaginalis cavity or intravenously was studied in 36 dogs. The dogs were divided into two equal groups: (1) an intratunical group in which 20 mg MXT solution was injected into the tunica vaginalis sac of each dog and (2) a parenteral group in which the same MXT dose was administered intravenously. The MXT concentration in serum, testicle and epididymis was determined 2, 4 and 24 h after MXT administration. Clinical and histologic examination of the tunica vaginalis was performed weekly for four consecutive weeks. The intratunical route, in contrast to the parenteral route, achieves a high MXT level in the testicle and epididymis with a low serum level, resulting in low toxicity and high drug efficacy. This route may therefore be suitable for a more effective treatment of testicular and epididymal lesions, notably malignant tumors.

Extended stability of 5-fluorouracil and methotrexate solutions in PVC containers

The extended physical and chemical stability of 5-fluorouracil and methotrexate in 0.9% sodium chloride injection in PVC containers was monitored for periods of up to 3 months and compared to solutions stored in glass containers. For 5-fluorouracil, 50 mg/ml solutions stored at room temperature are stable for up to 42 days and 5 mg/ml solutions for up to 13 weeks of refrigerated storage (5 ± 3°C) plus an additional week at room temperature. Methotrexate solutions at concentrations ranging from 1.25 to 12.5 mg/ml are stable for up to 15 weeks of refrigerated storage followed by an additional week at room temperature. No physical or chemical modifications of the solutions were observed, the only limiting factor for long-term stability being a progressive increase in drug concentration due to evaporative water loss from the PVC containers. Except for this slight increase in drug concentration, the same results were obtained for both plastic and glass containers.

A fluorescein-methotrexate-based flow cytometric bioassay for measurement of plasma methotrexate and trimetrexate levels

We describe a bioassay for the quantitation of plasma methotrexate and trimetrexate levels employing intact cells. This assay is based on the intracellular saturation of dihydrofolate reductase with fluorescein-methotrexate (F-MTX) and its dose-dependent displacement by methotrexate or trimetrexate as monitored by flow cytometry. Serially diluted methotrexate-containing plasma, representing a wide chemotherapeutic range, produces F-MTX displacement curves similar to those of standard methotrexate solutions. There is no interference by normal plasma components such as folate and its reduced forms. Plasma methotrexate or trimetrexate concentration is the product of the 50% displacing concentration of standard antifolate (IC 50) and the reciprocal of the plasma dilution which yields the same displacement. F-MTX competition with standard methotrexate displayed linear displacement from 18.0 ± 3.1 to 33.7 ± 1.5 n m ( n = 10). The standard trimetrexate calibration curve was linear from 0.28 ± 0.03 to 1.5 ± 0.33 n m ( n = 8). Thus, the bioassay sensitivities for methotrexate and trimetrexate are at least 18 and 0.3 n m, respectively. Comparison of methotrexate levels in 10 plasma specimens from cancer patients determined by the clinical enzyme inhibition assay and by our bioassay showed a high degree of correlation ( r = 0.987).

Stability of methotrexate injection in prefilled, plastic disposable syringes

The stability of methotrexate (MTX) injection in 4 commercially available brands of sealed plastic disposable syringe has been investigated. MTX injection at a concentration of 50 mg/ml or less, stored in sealed Monoject (Sherwood Medical) or Plastipak (Becton Dickinson), plastic disposable syringes in the absence of light at a temperature not exceeding 25 °C was found to be stable (less than 10% degradation) for a period of up to 8 months. A possible alteration in the permeability of the syringe material to water vapour on prolonged storage in contact with MTX solution precluded storage in Sabre (Gillette) or Steriseal (N.I. Ltd.) syringes for more than 70 days at 25°C. Sealed plastic disposable syringes are, therefore, a suitable secondary packaging system for the storage of MTX injection for periods of up to 8 months.

Methotrexate in infusion solutions--a stability test for the hospital pharmacy.

The stability of methotrexate solutions, subject to a freeze-thaw procedure, has been investigated. According to our results these solutions can be produced in batches in the hospital pharmacy and thawed to room temperature immediately before use without any degradation of methotrexate within the time limits given.

Effect of Polycation Complexation on Methotrexate-Liposome Cytotoxicity

AbstractMethotrexate and methotrexate-DEAE dextran complex were microencapsulated in positively charged liposomes. Their cytotoxicity was determined and compared with the cytotoxicity of control systems against L1210 mouse leukemia eel Is at 37°C In acetate buffer of pH 7.40 ± 0.05. The control systems used were acetate buffer, blank liposome, DEAE-dextran liposome and methotrexate solution. The methotrexate and methotrexate-DEAE dextran liposomes were lyophilized and the Influence of lyophilization on their cytotoxicity was also examined. The methotrexate-DEAE dextran liposomes resulted in slightly higher mean growth ratios than the free methotrexate liposomes in non lyophilized as well as lyophilized systems. The ED50 values for methotrexate and methotrexate-DEAE dextran liposomes were similar to that for the free methotrexate solution. This observation indicated that microencapsulation of methotrexate in liposomes either as a free drug or a complex has no effect on the cytotoxicity of the drug. In addi...

Evaluation of Some Pharmaceutical Aspects of Intrathecal Methotrexate Sodium, Cytarabine and Hydrocortisone Sodium Succinate

Methotrexate sodium, cytarabine and hydrocortisone sodium succinate were evaluated in three vehicles, Sodium Chloride Injection, USP, Lactated Ringer's Injection, USP, and Elliott's B solution, an artificial cerebrospinal fluid (CSF). Osmolarity values of all drug solutions studied were within 10% of normal human. No vehicle-related alterations in the stability of methotrexate or cytarabine were detected under simulated use conditions. However, the degradation of hydrocortisone sodium succinate was faster in Elliott's B solution than in the other vehicles. Principal differences among the vehicles involved electrolyte content, pH and buffer capacity. Elliott's B solution is similar to CSF in electrolyte composition and buffer capacity, and was the only vehicle to maintain the pH of dilute methotrexate and cytarabine solutions between 7.2 and 7.8. The significance of these in vitro data await a careful clinical study.

Factors Affecting Human Percutaneous Penetration Of Methotrexate And Its Analogues In Vitro

Factors affecting the percutaneous penetration of methotrexate (MTX) and its analogues through excised human skin were studied. With application of 0.05% MTX solution to the epidermis, 0.07% of the applied dose was recovered in the epidermis and less than 0.005% penetrated completely through the skin. The percent of drug which penetrated remained constant from 0.05% to 2.0% MTX and was also unchanged by increasing incubation temperature from 28 degrees C to 37 degrees C. Cellophane tape stripping of stratum corneum following drug incubation removed essentially all the MTX that had been measured in whole epidermis. Binding of MTX was localized to the outermost layers of the stratum corneum. With removal of the stratum corneum prior to drug incubation, 25% of the applied MTX penetrated through the skin. Various vehicles were tested to determine their effect on MTX percutaneous absorption. Dimethylsulfoxide (80%), dimethylacetamide (25%), or retinoic acid (0.1%) had no effect on penetration. Maximum enhancement of penetration by 25-fold and 143-fold was obtained with retinoic acid (saturated solution in aqueous ethanol) and C-10-methylsulfoxide (2.5%), respectively. The penetration of lipid-soluble derivatives of MTX was also tested. The compound that penetrated best was the dimethyl ester of dichloro-MTX with a 3-fold increase in penetration over MTX.