RESUMEN
Hematide is a synthetic PEGylated peptidic erythropoiesis stimulating agent (ESA) that is presently being developed for the correction of anemia in patients with chronic renal failure. Unlike currently marketed ESAs, Hematide does not possess any sequence homology to erythropoietin (EPO) and has not elicited moribund immune responses in animal safety studies thereby allowing the generation of a robust safety package. Animals administered marketed ESAs develop anti-EPO antibodies that null the effect of the administered ESA and neutralize endogenous EPO, resulting in severe anemia that precludes the interpretation of chronic safety studies. The primary objective of this study is to determine whether Hematide-specific antibodies are generated when male monkeys are exposed to high Hematide doses (10 mg/kg, intravenous [IV] and subcutaneous [SC]) administered at frequent dosing intervals (every two weeks) for a total of 9 doses; secondary objectives are to evaluate whether developed antibodies impact pharmacokinetics (PK) and pharmacology. In this study, no Hematide-specific antibodies were detected. Hematide exhibits a prolonged plasma half-life and slow clearance by either IV or SC administration. Hematide induced significant erythropoiesis with reticulocytosis and subsequent increases in red blood cells, hematocrit and hemoglobin (Hgb) levels. No erythropoietic differences were noted between the IV and the SC dosed groups with mean +/- SD Hgb levels of 20.9 +/- 2.5 and 20.3 +/- 2.1 g/dL, respectively, occurring on Day 48, corresponding to Hgb increases of 6.5 and 6.7 g/dL, respectively, over pre-dose levels. In conclusion, Hematide is a potent erythropoiesis stimulating agent that exhibits plasma persistence in monkeys. Similar erythropoietic responses were produced following IV and SC administration. The absence of antibody development suggests that Hematide, at the doses and regimen described, has a low immunogenic potential in cynomolgus monkeys.
Asunto(s)
Eritropoyesis/efectos de los fármacos , Péptidos/farmacología , Polietilenglicoles/farmacología , Animales , Eritropoyetina/farmacología , Hemoglobinas/análisis , Inyecciones Intravenosas , Inyecciones Subcutáneas , Macaca fascicularis , Masculino , Péptidos/administración & dosificación , Péptidos/inmunología , Péptidos/farmacocinética , Polietilenglicoles/administración & dosificación , Polietilenglicoles/farmacocinética , Proteínas Recombinantes , Reticulocitos/efectos de los fármacosRESUMEN
A covalent conjugate (NR-LU-10/SA) was prepared between streptavidin (SA) and NR-LU-10, a mAb that binds an antigen expressed on the surface of most human carcinomas. NR-LU-10/SA was injected into nude mice bearing human tumor xenografts. Injection of biotinylated galactosyl-human serum albumin reduced the circulating levels of conjugate by 95%. Subsequent administration of (90)Y-1,4,7, 10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-biotin achieved peak uptake at the tumor within 2 hr while >80% of the radioactivity was eliminated in the urine. A single dose of 600-800 microCi of (90)Y-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-biotin produced cures in 10/10 mice with established (>200 mm(3)) s.c. human small cell lung or colon cancer xenografts and 8/10 cures in mice with human breast cancer xenografts without significant toxicity.
Asunto(s)
Carcinoma/metabolismo , Neoplasias del Colon/tratamiento farmacológico , Trasplante de Neoplasias , Trasplante Heterólogo , Radioisótopos de Itrio , Radioisótopos de Itrio/uso terapéutico , Animales , Anticuerpos Monoclonales , Antígenos de Neoplasias/inmunología , Antineoplásicos/sangre , Antineoplásicos/farmacología , Moléculas de Adhesión Celular/inmunología , Quelantes/química , Neoplasias del Colon/inmunología , Molécula de Adhesión Celular Epitelial , Compuestos Heterocíclicos con 1 Anillo/química , Humanos , Ratones , Ratones Desnudos , Estructura Molecular , Radioinmunoterapia/métodos , Radiofármacos/uso terapéutico , Radiofármacos/toxicidad , Radioisótopos de Itrio/toxicidadRESUMEN
UNLABELLED: Pretargeted radioimmunotherapy (PRIT) was evaluated using an antibody-streptavidin conjugate, followed by a biotin-galactose-human serum albumin clearing agent and 90Y-dodecane tetraacetic acid (DOTA)-biotin as the final step for therapy. The objective was to develop a clinical protocol that could show an improved tumor-to-red marrow therapeutic ratio compared with conventional radioimmunotherapy (RIT) and at the same time preserve the efficiency of tumor targeting. METHOD: Forty-three patients with adenocarcinomas reactive to NR-LU-10 murine monoclonal antibody received the 3 components. Doses and timing parameters were varied to develop an optimized schema. In some patients, the conjugate was radiolabeled with 186Re as an imaging tracer to assess biodistribution of the conjugate and effectiveness of the clearing agent. 111In-DOTA-biotin was coinjected with 90Y-DOTA-biotin for quantitative imaging. Safety, biodistribution, pharmacokinetics, dosimetry, and antiglobulin formation were evaluated. RESULTS: The optimal schema was defined as a conjugate dose of 125 microg/mL plasma volume followed at 48 h by a clearing agent in a 10:1 molar ratio of clearing agent to serum conjugate. The therapeutic third step was 0.5 mg radiobiotin administered 24 h later. No significant adverse events were observed after administration of any of the components. The mean tumor-to-marrow absorbed dose ratio when using the optimized PRIT schema was 63:1, compared with a 6:1 ratio reported previously for conventional RIT. Antiglobulin to murine antibody and to streptavidin developed in most patients. CONCLUSION: This initial study confirmed that the PRIT approach is safe and feasible and achieved a higher therapeutic ratio than that achieved with conventional RIT using the same antibody.
Asunto(s)
Adenocarcinoma/radioterapia , Radioinmunoterapia/métodos , Radioisótopos de Itrio/uso terapéutico , Estudios de Factibilidad , Femenino , Humanos , Masculino , Factores de TiempoRESUMEN
A simplified kiet has been developed for 99mTc protein radiolabeling using an N3S triamide mercaptide bifunctional chelating agent and the preformed chelate approach. The process combined N3S chelating agent, gluconate intermediate transfer agent, stannous reducing agent, and gentisic acid stabilizer into a lyophilized formulation. With sulfur donor atom hemithioacetal protection of the ligand, delta-2,3,5,6-tetrafluorothiophenyl alpha-S-(1-ethoxyethyl)mercaptoacetamido-L-adipoylglycylglycine , optimum 99mTc chelation was achieved in a single step. Subsequent reaction with NR-LU-10 antibody Fab fragment followed by purification via QAE Sephadex anion exchange resin filter afforded 99mTc-N3S-NR-LU-10 Fab conjugate with retained immunoreactivity and effective tumor targeting properties.
Asunto(s)
Quelantes/química , Gentisatos , Marcaje Isotópico/métodos , Compuestos de Organotecnecio , Animales , Quelantes/farmacocinética , Neoplasias del Colon/metabolismo , Gluconatos , Hidroxibenzoatos , Fragmentos Fab de Inmunoglobulinas , Inmunotoxinas/química , Indicadores y Reactivos , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Compuestos de Organotecnecio/farmacocinética , Distribución TisularRESUMEN
Variant isoforms of CD44, a family of cell-surface glycoproteins generated by alternative splicing and post-translational modifications, are expressed in a variety of human tumors and play important roles in tumor progression and metastasis formation. The murine monoclonal IgG1 antibody VFF18, specific for an epitope encoded by human CD44 variant exon 6, binds with high affinity to the recombinant protein (Kd = 1.7 x 10(-10) M) as well as to tumor cell lines in vitro, and is suitable for immunohistochemical analysis of human tumors. Screening of more than 500 tumor samples of different histogenesis showed that VFF18 most strongly and uniformly reacts with squamous cell carcinomas (SCC). Detailed analysis of 185 SCC (head and neck, lung, skin) confirmed reactivity of the antibody with 99% of the samples, with intense and homogeneous staining of the tumor cells in the majority of cases, whereas reactivity of VFF18 with normal tissues is limited to certain epithelia and activated lymphocytes. When radiolabelled VFF18 was administered to nude mice bearing human epidermoid carcinoma (A-431) xenograft, fast and selective tumor uptake of the radioimmunoconjugate with a maximum of 18% of the injected dose per gram of tissue was observed. Taken together, these data suggest that mAb VFF18 is a promising targeting vehicle for radioimmunotherapy of squamous cell carcinomas in humans.
Asunto(s)
Anticuerpos Monoclonales/metabolismo , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/terapia , Receptores de Hialuranos/metabolismo , Neoplasias/metabolismo , Neoplasias/terapia , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacocinética , Especificidad de Anticuerpos , Carcinoma de Células Escamosas/inmunología , Epitelio/inmunología , Epitelio/metabolismo , Femenino , Humanos , Receptores de Hialuranos/inmunología , Inmunohistoquímica , Inmunoterapia , Radioisótopos de Yodo , Isomerismo , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Datos de Secuencia Molecular , Neoplasias/inmunología , Radioinmunodetección , Trasplante HeterólogoRESUMEN
A mouse-human chimeric monoclonal antibody (NR-LU-13), with the same pancarcinoma antigen recognition site as a previously studied murine monoclonal antibody (NR-LU-10), was radiolabeled with 186Re using a bifunctional chelate. Nine patients (ages 31-81 yr) with metastatic adenocarcinoma received 186Re NR-LU-13. A single intravenous dose of 42 mg NR-LU-13 labeled with 25 mCi/m2 (two patients) or 60 mCi/m2 (seven patients) was administered. Mean serum disappearance half-time values for the chimeric 186Re NR-LU-10). Fifty percent of the radiolabel was excreted in the urine by 6 days. Tumor localization was demonstrated by gamma camera imaging in seven of nine patients. The percent injected dose per gram in a single tumor biopsy specimen was 0.003% at 72 hr postinjection. Absorbed dose to bone marrow was 1.5 +/- 0.7 rads/mCi and resulted in reversible myelosuppression in five of six evaluable patients who received 60 mCi/m2: median WBC nadir = 2500/microliters; median platelet nadir = 85,500/microliters. Low grade fever, nausea, slight elevations of liver function tests and mild allergic reactions were seen in some patients. The chimeric antibody elicited low levels of anti-NR-LU-13 antibody in six of eight evaluable patients (75%), in contrast to NR-LU-10 which elicited higher levels of human anti-mouse antibody in all patients. This pilot study demonstrates the ability of the chimeric antibody to target tumors with reduced (but not absent) immunogenicity and delayed clearance relative to the murine antibody.
Asunto(s)
Anticuerpos Monoclonales/metabolismo , Radioinmunoterapia , Radioisótopos/farmacocinética , Proteínas Recombinantes de Fusión/farmacocinética , Renio/farmacocinética , Adenocarcinoma/diagnóstico por imagen , Adenocarcinoma/inmunología , Adenocarcinoma/metabolismo , Adenocarcinoma/radioterapia , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales/efectos adversos , Anticuerpos Monoclonales/inmunología , Anticuerpos Antineoplásicos/inmunología , Formación de Anticuerpos , Semivida , Humanos , Persona de Mediana Edad , Radioisótopos/efectos adversos , Cintigrafía , Proteínas Recombinantes de Fusión/inmunología , Renio/efectos adversos , Renio/inmunología , Distribución TisularRESUMEN
Rhenium-186 is a beta-emitting radionuclide that has been studied for applications in radioimmunotherapy. Its 137 keV gamma photon is ideal for imaging the biodistribution of the immunoconjugates and for obtaining gamma camera data for estimation of dosimetry. Methods used for determining radiation absorbed dose are described. We have estimated absorbed dose to normal organs and tumors following administration of two different 186Re-labeled immunoconjugates, intact NR-LU-10 antibody and the F(ab')2 fragment of NR-CO-02. Tumor dose estimates in 46 patients varied over a wide range, 0.4-18.6 rads/mCi, but were similar in both studies. Accuracy of activity estimates in superficial tumors was confirmed by biopsy. Prediction of 186Re dosimetry from a prior 99mTc imaging study using a tracer dose of antibody was attempted in the NR-CO-02 (Fab')2 study. Although 99mTc was an accurate predictor of tumor localization and the mean predicted and observed radiation absorbed doses to normal organs compared favorably, 186Re dosimetry could not be reliably predicted in individual patients. The methods described nevertheless provide adequate estimates of 186Re dosimetry to tumor and normal organs.
Asunto(s)
Radioinmunodetección , Radioinmunoterapia , Radioisótopos/uso terapéutico , Renio/uso terapéutico , Humanos , Radioisótopos de Yodo/uso terapéutico , Neoplasias Hepáticas/diagnóstico por imagen , Neoplasias Hepáticas/radioterapia , Neoplasias Pulmonares/diagnóstico por imagen , Neoplasias Pulmonares/radioterapia , Radiometría , Tecnecio , Distribución TisularRESUMEN
The variability in plasma and urine equine procaine measurement between three independent laboratories using current methods led to the development of a sensitive, reliable, and reproducible high-performance liquid chromatographic method. Standardbred mares were administered either a penicillin G procaine preparation intramuscularly or procaine hydrochloride subcutaneously, and blood and urine were collected at defined time intervals. By HPLC the detection limits for procaine in plasma and urine were 1 and 10 ng/mL, respectively. In contrast procaine in plasma could not be detected by GC-NPD, while the urinary detection limit was 50 ng/mL. The concentration of fluoride in the collection tubes and repetitive freeze-thawing modified plasma procaine measurement. Urinary pH was a factor in estimation of urine procaine levels with greater recovery and reproducibility of results at pH 5 as compared to pH 7. This HPLC method provides a simple, sensitive, and reliable quantitation of procaine in equine plasma and urine.
Asunto(s)
Cromatografía Líquida de Alta Presión/veterinaria , Caballos/metabolismo , Procaína/análisis , Animales , Doping en los Deportes , Procaína/sangre , Procaína/orinaRESUMEN
The Oyen et al. study is valuable in that it systematically evaluates several of the factors involved in radiolabeled protein uptake and retention in infectious foci. The role of particular proteins and their receptor specific interactions seems to be inconsequential in agreement with the findings of other. However, the role of the radiolabel was shown to be important and significant differences were delineated from comparisons of the radionuclides and their associated chemistries. The conclusion implicating radionuclide chemistry and associated linkages underscores the need to optimize the attachment and labeling chemical modifications of protein carriers. Evaluation criteria should include serum stability, determination and assessment of the effect of molar substitution ratio, and potential for improving blood clearance without reducing the target-to-non-target ratio. Important areas for future study include characterization of radioactive metabolites and the design and synthesis of new ligands which direct the disposition of metabolites reducing retention in normal organs or accelerating renal excretion. Additionally, intracellular processing of radiolabel, compartmental distribution and strategies for augmenting internalization and retention within the target cell merit detailed exploration. For each radionuclide of interest, 111In, radioiodines, 99mTc and others, improved chemical moieties exist for controlling radiolabel fate. When carrying out mechanistic and evaluative studies, clear-cut conclusions will only be reached when defined and controlled chemistry is used. Having established a "gold standard," simplifications in radiolabeling and other chemical refinements can then be pursued with a quantitative understanding of the trade-offs in targeting agent performance versus other considerations such as cost reduction, simplicity, and convenience.
Asunto(s)
Infecciones Bacterianas/metabolismo , Inmunoglobulina A/metabolismo , Inmunoglobulina G/metabolismo , Radioisótopos de Indio/farmacocinética , Radioisótopos de Yodo/farmacocinética , Albúmina Sérica/farmacocinética , Tecnecio/farmacocinética , Infecciones Bacterianas/diagnóstico por imagen , Cintigrafía , Distribución TisularRESUMEN
Plasma and urinary concentrations of procaine were examined in Standardbred mares after subcutaneous administration of various doses (80 mg to 1600 mg) of procaine hydrochloride. Regardless of dose, peak plasma procaine values occurred within 1 h, but remained detectable in a dose-dependent manner, with procaine present at 1 h with the 80 mg dose and 6 h at the 1600 mg dose. Similarly, peak urinary procaine concentrations were attained within 1.5 to 3 h, irrespective of dose, while detection time was dose-dependent, being 23 h for 80-200 mg doses but as long as 30-54 h with the 1600 mg dose. When mares were given a single intramuscular injection of a penicillin G-procaine preparation (Ethacillin, Cillimycin, Penamycin, Derapen A, Azimycin or Diathal), peak plasma procaine concentrations varied and were reached from 10 min to 3 h in all cases, with detection from 3 to 20 h after drug administration. Although the peak urinary levels of procaine occurred between 30 mins and 6 h, detection in urine in most cases was as long as 78-120 h except for Diathal for which detection was limited to 54 h. Daily administration of a penicillin G-procaine preparation (Pen-Di-Strep) for 5 days produced a biphasic peak in plasma procaine at 3 and at 6-9 h with detection from 16 to 23 h after drug treatment. Although peak urinary procaine values were reached at similar times after single or multiple injections, the duration of detection was markedly longer (425 h) after the multiple-dose regimen.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Doping en los Deportes , Caballos/metabolismo , Procaína/farmacocinética , Animales , Cruzamiento , Relación Dosis-Respuesta a Droga , Femenino , Inyecciones Intramusculares/veterinaria , Inyecciones Subcutáneas/veterinaria , Procaína/administración & dosificación , Procaína/sangre , Procaína/orinaRESUMEN
Glycopyrrolate (Robinul) is a quaternary ammonium salt which serves as a respiratory enhancing drug. It is reportedly used in horse racing to improve breathing. Extraction of glycopyrrolate from equine urine employing unique solid-phase extraction techniques gave a residue suitable for liquid chromatography-tandem mass spectrometry (LC-MS-MS) and gas chromatography-mass spectrometry (GC-MS). LC-MS-MS analysis employed an extract derived from 5 ml of urine subjected to cation-exchange chromatography. The daughter ion of m/z 318 monitored in the positive-ion mode was m/z 116. Recovery of glycopyrrolate was 99.5% and the within-run coefficient of variation of two quality control samples (1.0 and 10 ng/ml) was less than 5%. The between-run coefficient of variation for the same two quality control samples was less than 6.5%. The minimal detectable concentration for the assay was 250 pg/ml. Due to the extremely low concentration of glycopyrrolate in urine, qualitative detection via full-scan GC-MS required XAD-2 extraction of 50 ml of urine, cation-exchange chromatography clean-up and a tandem hydrolysis-derivatization procedure. The target analyte for GC-MS qualitative analysis was the methyl ester of hydrolyzed glycopyrrolate. Glycopyrrolate could be detected in post-administration (1 mg intravenously) urine samples for up to 9 h by both LC-MS-MS and GC-MS. The success of the method was due to a combination of the extreme sensitivity of the LC-MS-MS method and the very selective extraction process for quaternary ammonium salts.
Asunto(s)
Glicopirrolato/orina , Caballos/orina , Animales , Cromatografía por Intercambio Iónico , Cromatografía Liquida , Cromatografía de Gases y Espectrometría de Masas , Indicadores y Reactivos , Espectrometría de Masas , Estándares de ReferenciaRESUMEN
A kit has been developed for 99mTc antibody radiolabeling via defined chemistry using an N2S2 diamide dimercaptide bifunctional chelating agent and the performed chelate method. The process involved efficient transchelation of 99mTc from gluconate to 2,3,5,6-tetrafluorophenyl 4,5-bis-S-(1-ethoxyethyl) mercaptoacetamidopentanoate as an active ester ligand and subsequent conjugation to antibody lysine amine functional groups. The use of the ethoxyethyl group for sulfur protection allowed optimum yields of 99mTc N2S2 chelate formation with complete retention of the active ester. Subsequent addition of antibody Fab fragment gave 99mTc chelate conjugates indistinguishable from the stepwise in situ esterification and purification of the 99mTc N2S2 complex followed by conjugation as previously shown to give stable 99mTc antibody fragments with retained immunoreactivity and tumor-targeting properties.
Asunto(s)
Aminoácidos Diaminos , Aminoácidos Sulfúricos , Fragmentos Fab de Inmunoglobulinas , Juego de Reactivos para Diagnóstico , Tecnecio , Aminoácidos Diaminos/síntesis química , Aminoácidos Diaminos/farmacocinética , Aminoácidos Sulfúricos/síntesis química , Aminoácidos Sulfúricos/farmacocinética , Animales , Estudios de Evaluación como Asunto , Fragmentos de Inmunoglobulinas , Marcaje Isotópico , Ratones , Ratones Desnudos , Distribución TisularRESUMEN
A theoretical comparison was made of radioimmunotherapy (RIT) dosimetry estimates for eight radionuclides (90Y, 105Rh, 131I, 153Sm, 186Re, 188Re, 198Au, 211At) conjugated to IgG, F(ab')2, and Fab antibody forms. Antibody pharmacokinetics, derived from a nude mouse animal model were combined with appropriate physical data and S values to evaluate absorbed dose to a 0.5 kg centrally located tumor, total body and kidney. Radioimmunoconjugates of F(ab')2 with 90Y, 153Sm and 186Re were predicted to be the most promising for RIT.
Asunto(s)
Anticuerpos , Radioinmunoterapia , Radioisótopos , Animales , Anticuerpos Monoclonales/inmunología , Médula Ósea/metabolismo , Femenino , Semivida , Hematopoyesis/fisiología , Humanos , Fragmentos Fab de Inmunoglobulinas/inmunología , Inmunoglobulina G/inmunología , Riñón/metabolismo , Ratones , Ratones Desnudos , Modelos Biológicos , Trasplante de Neoplasias/inmunología , Radioisótopos/farmacocinéticaRESUMEN
A 186Re-labeled monoclonal antibody (MAb), NR-LU-10, was used for the radioimmunotherapy of a subcutaneous human small cell lung carcinoma xenograft, SHT-1, in nude mice. Biodistribution with specific and irrelevant labeled MAb demonstrated peak tumor uptake of 8% and 3% of the injected dose/g at 2 days, respectively. Dosimetry analysis predicted tumor:whole-body radiation-absorbed dose ratios of 2.43:1 for NR-LU-10 and 0.62:1 for irrelevant MAb. Single-dose toxicity screening estimated a 50% lethal dose within 30 days of 600 microCi (880 cGy of whole-body radiation). As anticipated, a multiple-dose regimen of 490 microCi in four doses over 10 days (720 cGy of whole-body radiation, eight of eight surviving greater than 30 days) was less toxic than a single bolus dose of 430 microCi (644 cGy of whole-body radiation), six of eight surviving greater than 30 days). A multidose radioimmunotherapy regimen was initiated in nude mice bearing 66-mm3 tumors (total dose, 500 to 600 microCi). Complete remissions (greater than 140 days) were achieved in three of 16 mice, and the remainder showed a mean tumor growth delay of 53 days. Matched doses with irrelevant MAb produced one remission, one treatment-related death, and a mean growth delay of only 20 days in six of eight mice. Thus, in this nonoptimal radioimmunotherapy model, significant antitumor responses were observed using a mildly toxic multiple dosing regimen.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Carcinoma de Células Pequeñas/terapia , Neoplasias Pulmonares/terapia , Radioisótopos/uso terapéutico , Renio/uso terapéutico , Animales , Autorradiografía , Carcinoma de Células Pequeñas/patología , Carcinoma de Células Pequeñas/radioterapia , Línea Celular , Femenino , Humanos , Inmunoterapia , Neoplasias Pulmonares/patología , Neoplasias Pulmonares/radioterapia , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Renio/farmacocinética , Distribución Tisular , Trasplante HeterólogoRESUMEN
A study of the effects of intravenous administration of either 150 mg or 250 mg of furosemide to standardbred mares pre-treated with other drugs was undertaken to determine whether a unique pattern of drug elimination into urine and from plasma for each compound occurred. Furosemide significantly reduced the plasma concentrations of codeine compared to control 2-6 h after furosemide administration. In contrast, the plasma concentrations of theophylline, phenylbutazone, pentazocine, guaifenesin and flunixin were not markedly altered by furosemide. In the case of acepromazine, clenbuterol and fentanyl, the data generated were insufficient to state with certainty whether or not furosemide affected the plasma concentrations of these three drugs. A significant reduction was noted in the urinary concentrations of guaifenesin, acepromazine, clenbuterol, phenylbutazone, flunixin, fentanyl and pentazocine within 1-4 h of furosemide administration. The urinary concentrations of theophylline remained reduced as long as 8 h after furosemide injection. Furosemide administration to horses pre-treated with codeine resulted in depression of urinary morphine concentrations 2-4 h and 9-12 h after furosemide injection. A lower furosemide dose (150 mg) produced changes in drug urinary excretion and plasma elimination equivalent to the higher dose (250 mg). It is evident that furosemide affects the urinary and plasma concentrations of other co-administered drugs but not in a predictable fashion, which limits the extrapolation of these results to as yet untested drugs.
Asunto(s)
Furosemida/farmacología , Caballos/metabolismo , Premedicación/veterinaria , Acepromazina/metabolismo , Animales , Clenbuterol/metabolismo , Clonixina/análogos & derivados , Clonixina/metabolismo , Codeína/metabolismo , Interacciones Farmacológicas , Femenino , Fentanilo/metabolismo , Furosemida/administración & dosificación , Caballos/orina , Inyecciones Intravenosas/veterinaria , Morfina/metabolismo , Pentazocina/metabolismo , Fenilbutazona/metabolismo , Esfuerzo Físico , Teofilina/metabolismoRESUMEN
Conjugates of monoclonal antibodies and Pseudomonas exotoxin A (PE) were formed with disulfide or thioether bonds. Thioether conjugates which formed with succinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (SMCC) modified PE and reduced antibody formed with an 80% yield of equimolar conjugate within 30 min with an offering of one to one (toxin:antibody). The efficiency and kinetics of thioether formation were much higher with SMCC than with other maleimide reagents as well as more efficient than disulfide linkers. Thioether linkage resulted in immunotoxin consistently more potent and more selective in vitro than disulfide bonded conjugate. Thioether bonded conjugates also proved to have other favorable in vivo properties compared to disulfide conjugates: (1) a longer half-life in serum; (2) increased tumor localization; and (3) reduced toxicity. Toxicity of thioether linked holotoxin conjugates was directed at the liver hepatocyte but was easily monitored by serum liver enzymes. The conjugates are currently undergoing clinical evaluation for treatment of ovarian cancer with intraperitoneal administration. Research is ongoing to further decrease residual toxicity without reducing the potency of the conjugate.
Asunto(s)
ADP Ribosa Transferasas , Toxinas Bacterianas , Exotoxinas/metabolismo , Inmunotoxinas/inmunología , Factores de Virulencia , Anticuerpos Monoclonales/metabolismo , Línea Celular , Fenómenos Químicos , Química , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Exotoxinas/toxicidad , Humanos , Inmunotoxinas/metabolismo , Inmunotoxinas/farmacología , Células Tumorales Cultivadas/patología , Exotoxina A de Pseudomonas aeruginosaAsunto(s)
Anticuerpos Antineoplásicos/uso terapéutico , Inmunoterapia , Melanoma/terapia , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/uso terapéutico , Antígenos de Neoplasias/inmunología , Diseño de Fármacos , Evaluación de Medicamentos , Ensayos de Selección de Medicamentos Antitumorales , Femenino , Gangliósidos/inmunología , Humanos , Fragmentos Fab de Inmunoglobulinas/uso terapéutico , Masculino , Melanoma/diagnóstico por imagen , Melanoma/inmunología , Melanoma Experimental/diagnóstico por imagen , Melanoma Experimental/inmunología , Melanoma Experimental/terapia , Antígenos Específicos del Melanoma , Ratones , Ratones Desnudos , Proteínas de Neoplasias/inmunología , Proteoglicanos/inmunología , Cintigrafía , Trasplante HeterólogoRESUMEN
L6 is a murine monoclonal antibody (MAb) binding to cells of most human carcinomas, mediating antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity, and inhibiting tumor growth in nude mice [10]. Fab and F(ab')2 fragments of L6, as well as intact MAb, have been evaluated for immunospecific localization in nude mice xenografted with a human lung carcinoma. They were compared with preparations of an isotype-matched control immunoglobulin, P1.17, after labelling with 125I or 131I. L6 Fab fragments prepared from MAb L6 and labelled with 67Ga via desferrioxamine were also tested. The data suggest that MAb L6 may be useful for in vivo detection of human carcinomas.
Asunto(s)
Anticuerpos Monoclonales , Neoplasias Experimentales/metabolismo , Animales , Femenino , Radioisótopos de Galio , Fragmentos Fab de Inmunoglobulinas , Radioisótopos de Yodo , Tasa de Depuración Metabólica , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Neoplasias , Neoplasias Experimentales/inmunología , Distribución TisularRESUMEN
A method of radioiodinating monoclonal antibodies such that the labeled antibodies do not undergo in vivo deiodination has been studied. The method utilizes conjugation of succinimidyl para-iodobenzoate to the antibody. The iodobenzoate was radiolabeled by using an organometallic intermediate to facilitate the reaction. Thus, succinimidyl para-tri-n-butylstannylbenzoate was radiolabeled in 60-90% radiochemical yield and subsequently conjugated to the antibody in 80-90% yield. Animal biodistribution studies were carried out with two separate anti-melanoma antibodies (9.2.27 and NR-M1-05) labeled by this method, and examined in nude mice bearing human melanoma tumor xenografts. Very large differences in the localization of radioactivity were observed in the thyroids and stomachs of mice when the iodobenzoyl-labeled antibodies were compared with the same antibodies labeled using the chloramine-T method of radioiodination. Few other significant differences in the tissue distribution of the radioiodinated antibodies were seen.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Radioisótopos de Yodo/uso terapéutico , Marcaje Isotópico/métodos , Neoplasias/radioterapia , Animales , Estabilidad de Medicamentos , Humanos , Indicadores y Reactivos , Melanoma/radioterapia , Ratones , Ratones Desnudos , Trasplante de Neoplasias , Trasplante HeterólogoRESUMEN
The data presented demonstrate that a human-murine chimeric antibody has been generated that retains the immunoreactivity and has similar pharmacokinetic properties of its parent murine monoclonal antibody NRML-05. A competitive ELISA assay demonstrated that antigen reactivity of both parent and chimera were nearly identical. In a direct cell binding assay, NRML-05 and chimeric antibodies were immunoreactive, 81% and 83%, respectively. Scatchard Analysis of the antibodies indicate very similar affinities for NRML-05 (4.4 x 10(9) M-1) and chimera (2.7 x 10(9) M-1). Localization of the two antibodies to tumor xenografts in nude mice were very similar in biodistribution studies. The chimeric antibody is not as well recognized by antiglobulin from patients who have responded to non-idiotypic murine antibody determinants. Although this data does not predict how immunogenic a chimera would be in the clinical setting, it does suggest that patients without significant anti-idiotypic antiglobulin response could benefit from second and subsequent administrations with chimeric antibody. Even though anti-idiotypic responses may occur with the chimera, these may be reduced as a result of the presentation of these epitopes on the less immunogenic human constant domains.