RESUMEN
The Bowman-Birk inhibitor (BBI) found in soybeans is a serine protease inhibitor with anticarcinogenic activity. In the present study, an ELISA for BBI was developed with the use of a monoclonal antibody against a reduced form of BBI. This newly developed ELISA method was used to measure the urinary levels of BBI metabolites in nine human subjects after consumption of 36-oz or 60-oz soymilk (containing 105 or 175 mg of BBI) at two time points 36 h apart. The results demonstrate that urinary BBI excretion rates peaked within 6 h and decreased to baseline levels within 12-24 h after soymilk ingestion. The changes in BBI:creatinine ratios in urine closely paralleled the changes in urinary BBI excretion rates after soymilk consumption. These data suggest that BBI ingested p.o. is absorbed and could be bioavailable for cancer chemoprevention in other organs in addition to those in the gastrointestinal tract.
Asunto(s)
Glycine max/química , Inhibidor de la Tripsina de Soja de Bowman-Birk/orina , Inhibidores de Tripsina/orina , Adulto , Anticuerpos Monoclonales , Disponibilidad Biológica , Creatinina/orina , Dieta , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Inhibidor de la Tripsina de Soja de Bowman-Birk/farmacocinética , Inhibidores de Tripsina/farmacocinéticaRESUMEN
The Bowman-Birk inhibitor (BBI) is a soybean-derived serine protease inhibitor and a potential cancer chemopreventive agent for humans. In this Phase I clinical trial, BBI concentrate was administered as a single oral dose to 24 subjects with oral leukoplakia. Pharmacokinetics of BBI was analyzed, and subjects were monitored clinically for toxic effects. Subjects received between 25 and 800 chymotrypsin inhibitor units (CIU) of the compound in a dose escalation trial. BBI was taken up rapidly, and a metabolic product of BBI was excreted in the urine within 24-48 h. No clinical or laboratory evidence of toxicity was observed in the study. Protease activity was also measured in buccal cells to evaluate usefulness as a biomarker. Single-dose BBI concentrate administered up to 800 CIU was well tolerated and appeared to be nontoxic. Further investigation in Phase II clinical trials is being done.
Asunto(s)
Anticarcinógenos/uso terapéutico , Leucoplasia Bucal/tratamiento farmacológico , Inhibidor de la Tripsina de Soja de Bowman-Birk/uso terapéutico , Inhibidores de Tripsina/uso terapéutico , Administración Oral , Anciano , Anticarcinógenos/administración & dosificación , Anticarcinógenos/efectos adversos , Anticarcinógenos/farmacocinética , Anticarcinógenos/orina , Biomarcadores/análisis , Quimioprevención , Quimotripsina/antagonistas & inhibidores , Endopeptidasas/análisis , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Mucosa Bucal/enzimología , Inhibidor de la Tripsina de Soja de Bowman-Birk/administración & dosificación , Inhibidor de la Tripsina de Soja de Bowman-Birk/efectos adversos , Inhibidor de la Tripsina de Soja de Bowman-Birk/farmacocinética , Inhibidor de la Tripsina de Soja de Bowman-Birk/orina , Inhibidores de Tripsina/administración & dosificación , Inhibidores de Tripsina/efectos adversos , Inhibidores de Tripsina/farmacocinética , Inhibidores de Tripsina/orinaRESUMEN
Bowman-Birk protease inhibitor (BBI) is a potent anticarcinogen that suppresses malignant transformation at nanomolar concentrations. Small amounts of BBI in its native form can be measured by immunoassay using specific monoclonal antibodies (MAbs); however, the MAbs currently available are not capable of detecting BBI metabolites in human body fluids. To develop new reagents for the study of BBI exposure and pharmacokinetics, we produced four MAbs, designated 3B6, 3E3, 4H8 and 5G2, from hybridomas derived from a mouse immunized with reductively modified BBI. The epitopes recognized by the four MAbs were characterized using BBI in its native form or modified by different methods. MAb 3B6 reacted with native BBI. Partial reduction of BBI with 720 Gy of gamma radiation in an oxygen-free solution of 100 mM formate increased the reactivity of BBI with 3B6; however, extensive reduction of BBI with 100 mM DL-dithiothreitol (DTT) completely abolished this antigenic reactivity. In contrast, the other three MAbs reacted with BBI molecules that had been reduced either with 720 Gy of radiation in formate solution or with DTT. Alkylation of the radiochemically reduced BBI with N-ethylmaleimide further increased the reactivity of BBI with 3E3, 4H8 and 5G2, possibly by preventing the formation of new disulfide bonds within the BBI molecules. The binding of 4H8 and 5G2 to BBI antigen was inhibited by the binding of 3E3, and vice versa. Thus, the epitopes recognized by 3E3, 4H8 and 5G2 are probably located close to one another on the reduced BBI molecules. These three MAbs were able to react with BBI metabolites in urine samples collected from volunteers after oral administration of BBI. The ability of these MAbs to detect BBI metabolites indicates that BBI may be reductively modified in vivo and these MAbs may be useful reagents for monitoring the uptake of BBI into human tissues in cancer chemoprevention studies with BBI.
Asunto(s)
Anticuerpos Monoclonales/análisis , Inhibidor de la Tripsina de Soja de Bowman-Birk/inmunología , Animales , Anticuerpos Monoclonales/biosíntesis , Western Blotting , Ensayo de Inmunoadsorción Enzimática , Epítopos/inmunología , Humanos , Hibridomas , Ratones , Ratones Endogámicos C57BL , Inhibidor de la Tripsina de Soja de Bowman-Birk/efectos de la radiación , Inhibidor de la Tripsina de Soja de Bowman-Birk/orinaRESUMEN
In this study, the distribution of the soybean-derived Bowman Birk inhibitor (BBI) in mice was examined. Mice received [125I]BBI by oral gavage and three hours later, the mice were sacrificed, the organs of interest were carefully removed and the distribution of the inhibitor was determined. The bulk of labeled BBI was present in the luminal contents of the small and large bowel, urine and feces. Significant amounts of label were also observed in the serum, esophagus, stomach and intestine, kidney, liver and lung. Analysis of tissue homogenates by gel filtration chromatography revealed that the radioactivity eluted from the column at the same position as the BBI standard indicating that the iodinated BBI was still intact. The chromatographically purified BBI was able to inhibit chymotrypsin indicating that functional protease inhibitory activity was present. These results indicate that the BBI becomes widely distributed in mice 3 h after oral administration and that intact protease inhibitor is present in internal organs.
Asunto(s)
Inhibidor de la Tripsina de Soja de Bowman-Birk/farmacocinética , Animales , Cromatografía en Gel , Quimotripsina/antagonistas & inhibidores , Heces/química , Intestino Grueso/química , Intestino Delgado/química , Radioisótopos de Yodo , Ratones , Distribución Tisular , Inhibidor de la Tripsina de Soja de Bowman-Birk/administración & dosificación , Inhibidor de la Tripsina de Soja de Bowman-Birk/orinaRESUMEN
BACKGROUND: The Bowman-Birk inhibitor is a soybean-derived protease inhibitor that has anti-inflammatory and anticarcinogenic activities. METHODS: A Phase I trial of Bowman-Birk inhibitor concentrate (BBIC) in 19 male subjects with benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTS) has been performed. RESULTS: The results of the trial indicated that there was no dose-limiting toxicity of BBIC. There was a statistically significant decrease in serum PSA levels in all BBIC-treated patients. Some BBIC-treated patients exhibited a relatively large reduction in serum PSA levels, ranging up to a 43% reduction. There was also a statistically significant decrease in serum triglyceride levels and a decrease in prostate volume in the treated patients. The scores recorded in response to a urinary symptom questionnaire indicated improved urinary activities in the BBIC-treated patients; however, the control subjects exhibited similar improvements in urinary activities during the course of the trial. CONCLUSIONS: The data obtained in this trial, particularly the data suggesting that BBIC treatment may lead to reduced serum PSA levels and reduced prostate volumes, suggest that a Phase II clinical trial of BBIC for the therapy of BPH is warranted.