Environmental Exposure
Conditions
Keywords
Benzo[a]pyrene, Accelerator Mass Spectrometry, Polycyclic Aromatic Hydrocarbons, cruciferous vegetable, Brussels sprouts, 3,3'-diindolylmethane, DIM, indole-3-carbinol, I3C, PAH
Brief summary
Evaluation of the pharmacokinetics for \[14C\]-benzo\[a\]pyrene (\[14C\]-BaP) and metabolites in plasma and urine over 48 hours following a 50 ng dose (5.4 nCi) alone, following 7 days' consumption of Brussels sprouts, and following 7 days' consumption of a supplement containing 3,3'-diindolylmethane (DIM).
Detailed description
The pharmacokinetics for \[14C\]-BaP and metabolites will be assessed by UHLPC-Accelerator Mass Spectrometry (AMS, Lawrence Livermore National Laboratory) in plasma and urine collected over 48 hours following oral doses of 50 ng dose (5.4 nCi) alone, following 7 days' consumption of Brussels sprouts, and following 7 days' consumption of a supplement containing 3,3'-diindolylmethane (DIM). The investigators hypothesize that pre-administration of Brussels sprouts or DIM will alter \[14C\]-BaP metabolism and increase the rate of elimination consistent with predictions based on a previously developed Physiologically-Based Pharmacokinetic (PBPK) model for BaP. Briefly, this hypothesis will be tested by dosing individuals with 50 ng \[14C\]-BaP alone and, following a 3-week washout period, ingestion of about 50 g Brussels sprouts or 300 mg of 3,3'-diindolylmethane (DIM) supplement for 7 days prior to the \[14C\]-BaP micro-dose. The impact of the supplement and the whole food will be assessed with respect to alterations in uptake from the GI tract, metabolism and rate of elimination. The consumption of cruciferous vegetables will be assessed at the beginning of the study by completion of a dietary questionnaire to examine typical eating patterns in the previous 3 months and by collection and extraction of blood and urine to assay for DIM by LC/ESI-MS/MS-SRM). In addition, for each phase, urine will be assayed for DIM as an estimate of crucifer or DIM supplement intake. In preclinical and clinical studies, administration of Brussels sprouts or DIM impacts the activity of the same enzymes responsible for the phase 1 (CYP1A1 and CYP1B1) and phase 2 enzymes (GSTM1, UGT, SULT). Monitoring changes in β-estradiol metabolites will confirm the mechanism of alteration in the metabolic profile of \[14C\]-BaP. Metabolite profiles and kinetics of elimination are predicted to be consistent with a BaP physiologically based pharmacokinetic (PBPK) model developed by Pacific Northwest National Laboratory (PNNL). A non-smoker, not exposed occupationally, receives 270-700 ng of BaP daily; about 95% dietary. The WHO has set an estimated safe daily lifetime (70 year/70 Kg individual, cancer endpoint) exposure to BaP of 42-350 ng. This protocol represents de minimus risk.
Interventions
Oral micro-dose (50 ng) (5.4 nCi)
DRUGBrussels sprouts before 50 ng dose
Brussels sprouts for 7 days before 50 ng (5.4 nCi) dose of BaP
DRUGDIM supplement before 50 ng dose
DIM supplement for 7 days before 50 ng (5.4 nCi) dose of BaP and coadministration with DIM supplement
Sponsors
National Institute of Environmental Health Sciences (NIEHS)
Lawrence Livermore National Laboratory
Pacific Northwest National Laboratory
Oregon State University
Study design
Allocation
NA
Intervention model
SINGLE_GROUP
Primary purpose
BASIC_SCIENCE
Masking
NONE
Masking description
Deidentified samples will be analyzed by AMS at Lawrence Livermore National Laboratory and the pharmacokinetics determine at Pacific Northwest National Laboratory.
Eligibility
Sex/Gender
ALL
Age
21 Years to 65 Years
Healthy volunteers
Yes
Inclusion criteria
* Age 21-65 (inclusive) * If female, must be post-menopausal or have had surgical sterilization to eliminate any possibility for fetal exposure * Willing to defer blood donation for one month before, throughout, and one month after completion of study activities * Willing to avoid consuming cruciferous vegetables, I3C or DIM supplements, smoked or cured meat or cheeses, or charcoal-grilled meats for 2 weeks prior to and during each study cycle (gas grilled foods acceptable) * Health history review and physical assessment showing general good health, as determined by study physician. Acceptable physical exam may have been conducted as part of protocol 8233 or 8554 if subject has not had significant changes in health status.
Exclusion criteria
* Smoker (tobacco or other substances) or use of smokeless tobacco in past 3 months or living with smoker * Regular use of medications that affect gut motility or nutrient absorption (e.g. cholestyramine, sucralfate, orlistat, pro- or anti-motility agents) * History of gastrointestinal surgery (e.g. bariatric surgery, cholecystectomy) or gastrointestinal disorder (Crohn's disease, celiac disease, IBS, or colitis) * Current or history of kidney or liver disease * Prior high-dose 14C exposure from medical tests. (micro-dose 14C exposure not exclusionary) * Occupational PAH exposure (e.g. roofers, asphalt pavers, fire-fighters, etc.) * Regular use of indole-3-carbinol or DIM dietary supplements * Allergy or intolerance to Brussels sprouts or similar foods
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Peak Plasma Concentration of 14C-BaP Cmax | 0-48 hours for each of 3 dosing cycles, with a washout period of 3 weeks between each dosing cycle | Determination of highest concentration of 14C-BaP in plasma. Blood samples collected at 0 (baseline), 0.25, 0.5, 1, 2, 3, 4, 8, 24, and 48 hour after dosing. All time points were used to determine Cmax. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Time at Highest Plasma Concentration of 14C-BaP Tmax | 0-48 hours for each of 3 dosing cycles, with a washout period of 3 weeks between each dosing cycle | Determination of time at which plasma concentration of 14C-BaP is highest. Blood samples collected at 0 (baseline), 0.25, 0.5, 1, 2, 3, 4, 8, 24, and 48 hour after dosing. All time points were used to determine Tmax. |
| Area Under Plasma Concentration of 14C-BaP Versus Time Curve AUC | 0-48 hours for each of 3 dosing cycles, with a washout period of 3 weeks between each dosing cycle | Integration of concentration of 14C-BaP in plasma over time. Blood samples collected at 0 (baseline), 0.25, 0.5, 1, 2, 3, 4, 8, 24, and 48 hour after dosing. All time points were used to determine AUC. |
| Rate of Elimination of 14C-BaP (Half Life) | 0-48 hours for each of 3 dosing cycles, with a washout period of 3 weeks between each dosing cycle | Determination of constants for rate of elimination of 14C-BaP from plasma. Blood samples collected at 0 (baseline), 0.25, 0.5, 1, 2, 3, 4, 8, 24, and 48 hour after dosing. All time points were used to determine half-life. |
Countries
United States
Participant flow
Participants by arm
| Arm | Count |
|---|---|
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose Cycle 1: Capsule containing 50 ng (5.4 nCi) \[14C\]-benzo\[a\]pyrene (BaP).
Cycle 2: Subjects will consume 50 g (about 1/2 cup) of lightly steamed Brussels sprouts each evening for 7 days prior to taking capsule containing 50 ng (5.4 nCi) \[14C\]-benzo\[a\]pyrene (BaP).
Cycle 3: Subjects will consume 300 mg DIM supplement ( 2 capsules of BioResponse DIM® 150) each evening for 7 days prior to taking capsule containing 50 ng (5.4 nCi) \[14C\]-benzo\[a\]pyrene (BaP). A 300 mg DIM dose will be co-administrated with the 50 ng BaP dose.
At least 3 weeks will pass between cycles as a washout period.
\[14C\]-benzo\[a\]pyrene: Oral micro-dose (50 ng) (5.4 nCi)
Brussels sprouts before 50 ng dose: Brussels sprouts for 7 days before 50 ng (5.4 nCi) dose of BaP
DIM supplement before 50 ng dose: DIM supplement for 7 days before 50 ng (5.4 nCi) dose of BaP and coadministration with DIM supplement | 7 |
| Total | 7 |
Baseline characteristics
| Characteristic | 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose |
|---|---|
| Age, Categorical <=18 years | 0 Participants |
| Age, Categorical >=65 years | 0 Participants |
| Age, Categorical Between 18 and 65 years | 7 Participants |
| Ethnicity (NIH/OMB) Hispanic or Latino | 0 Participants |
| Ethnicity (NIH/OMB) Not Hispanic or Latino | 7 Participants |
| Ethnicity (NIH/OMB) Unknown or Not Reported | 0 Participants |
| Race (NIH/OMB) American Indian or Alaska Native | 0 Participants |
| Race (NIH/OMB) Asian | 0 Participants |
| Race (NIH/OMB) Black or African American | 1 Participants |
| Race (NIH/OMB) More than one race | 0 Participants |
| Race (NIH/OMB) Native Hawaiian or Other Pacific Islander | 0 Participants |
| Race (NIH/OMB) Unknown or Not Reported | 0 Participants |
| Race (NIH/OMB) White | 6 Participants |
| Region of Enrollment United States | 7 participants |
| Sex: Female, Male Female | 1 Participants |
| Sex: Female, Male Male | 6 Participants |
Adverse events
| Event type | EG000 affected / at risk |
|---|---|
| deaths Total, all-cause mortality | 0 / 7 |
| other Total, other adverse events | 0 / 7 |
| serious Total, serious adverse events | 0 / 7 |
Outcome results
Primary
Peak Plasma Concentration of 14C-BaP Cmax
Determination of highest concentration of 14C-BaP in plasma. Blood samples collected at 0 (baseline), 0.25, 0.5, 1, 2, 3, 4, 8, 24, and 48 hour after dosing. All time points were used to determine Cmax.
Time frame: 0-48 hours for each of 3 dosing cycles, with a washout period of 3 weeks between each dosing cycle
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Peak Plasma Concentration of 14C-BaP Cmax | 50 ng [14C] BaP | 3.69 fg [14C]-BaP/mL plasma | Standard Deviation 4.12 |
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Peak Plasma Concentration of 14C-BaP Cmax | Brussels sprouts before 50 ng [14C] BaP | 1.67 fg [14C]-BaP/mL plasma | Standard Deviation 1.03 |
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Peak Plasma Concentration of 14C-BaP Cmax | DIM supplement before 50 ng [14C] BaP | 0.68 fg [14C]-BaP/mL plasma | Standard Deviation 0.51 |
Secondary
Area Under Plasma Concentration of 14C-BaP Versus Time Curve AUC
Integration of concentration of 14C-BaP in plasma over time. Blood samples collected at 0 (baseline), 0.25, 0.5, 1, 2, 3, 4, 8, 24, and 48 hour after dosing. All time points were used to determine AUC.
Time frame: 0-48 hours for each of 3 dosing cycles, with a washout period of 3 weeks between each dosing cycle
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Area Under Plasma Concentration of 14C-BaP Versus Time Curve AUC | 50 ng [14C] BaP | 10.93 fg [14C]-BaP/mL plasma x hour | Standard Deviation 9.19 |
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Area Under Plasma Concentration of 14C-BaP Versus Time Curve AUC | Brussels sprouts before 50 ng [14C] BaP | 9.34 fg [14C]-BaP/mL plasma x hour | Standard Deviation 10.02 |
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Area Under Plasma Concentration of 14C-BaP Versus Time Curve AUC | DIM supplement before 50 ng [14C] BaP | 5.51 fg [14C]-BaP/mL plasma x hour | Standard Deviation 6.17 |
Secondary
Rate of Elimination of 14C-BaP (Half Life)
Determination of constants for rate of elimination of 14C-BaP from plasma. Blood samples collected at 0 (baseline), 0.25, 0.5, 1, 2, 3, 4, 8, 24, and 48 hour after dosing. All time points were used to determine half-life.
Time frame: 0-48 hours for each of 3 dosing cycles, with a washout period of 3 weeks between each dosing cycle
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Rate of Elimination of 14C-BaP (Half Life) | 50 ng [14C] BaP | 3.69 hour | Standard Deviation 3.8 |
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Rate of Elimination of 14C-BaP (Half Life) | Brussels sprouts before 50 ng [14C] BaP | 3.86 hour | Standard Deviation 4.62 |
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Rate of Elimination of 14C-BaP (Half Life) | DIM supplement before 50 ng [14C] BaP | 9.75 hour | Standard Deviation 13.25 |
Secondary
Time at Highest Plasma Concentration of 14C-BaP Tmax
Determination of time at which plasma concentration of 14C-BaP is highest. Blood samples collected at 0 (baseline), 0.25, 0.5, 1, 2, 3, 4, 8, 24, and 48 hour after dosing. All time points were used to determine Tmax.
Time frame: 0-48 hours for each of 3 dosing cycles, with a washout period of 3 weeks between each dosing cycle
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Time at Highest Plasma Concentration of 14C-BaP Tmax | 50 ng [14C] BaP | 1.58 hour | Standard Deviation 1.16 |
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Time at Highest Plasma Concentration of 14C-BaP Tmax | Brussels sprouts before 50 ng [14C] BaP | 1.14 hour | Standard Deviation 0.9 |
| 50 ng Dose; Brussels Sprouts Before 50 ng Dose; DIM Supplement Before 50 ng Dose | Time at Highest Plasma Concentration of 14C-BaP Tmax | DIM supplement before | 14.00 hour | Standard Deviation 19.17 |