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Relative bioavailability of different DIM formulations

Relative bioavailability of different DIM formulations

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3,3'-Diindolylmethane (DIM) is known as an agent of natural origin that provides protection against different cancers due to the broad spectrum of its biological activities in vivo. However, this substance has a very poor biodistribution and absorption in animal tissues. This preclinical trial was conducted to evaluate the pharmacokinetics and bioa...

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... addition to the main pharmacokinetic parameters, relative DIM bioavailability, change in the relative bioavail- ability, and absolute DIM bioavailability for each substance have been estimated (Table 1). ...

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... Therefore, the dosing interval in this study was greater than the elimination half-life reported, and perhaps the plasma levels of DIM did not achieve the stationary state for obtaining an optimal therapeutic response. Some authors have established experimentally that plasma levels of DIM must be significantly more than 200 ng/ mL to achieve the therapeutic response; some other studies show that for a single dose, these plasma levels are reached with a dose of 200 mg of DIM-BR® (35). ...
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The Estrogen Metabolites (2-hydroxyestrogens: 16α-hydroxyestrone) Urine Ratio (EMUR) has been negatively associated with breast cancer; Mexican women have a lower EMUR than other populations. We evaluated the effectiveness of 3,3′-diindolylmethane (DIM) supplementation on increasing EMUR in premenopausal women. A randomized, double-blind clinical trial (NCT02525159 at ClinicalTrial.gov) was carried out on 60 women with an EMUR below 0.9. Patients were assigned randomly to receive a placebo or 75 mg of DIM a day (administered as 300 mg of DIM-BR®) for 30 day. Urine samples were obtained at baseline, at 30 day of supplementation, and 30 day after finishing supplementation. A Mann-Whitney U test was used to compare the EMUR; an ANOVA was used to analyze differences in body composition. EMUR was analyzed using ESTRAMET™ kits. While DIM-treated subjects did not increase their EMUR at 30 day of supplementation (p > 0.05), there was a non-significant positive trend 30 day once supplementation ended (p = 0.06). The DIM group saw a more significant decrease in body fat percentage than the placebo group (p = 0.04). In premenopausal Mexican women, 75 mg of the daily DIM supplement was ineffective in increasing EMUR; further studies are needed to evaluate the effective dosage, time frames, and effect on body fat.
... Hauder et al. (2011) determined the LOD and LOQ values for DIM in human plasma and urine samples by HPLC-MS/MS to be 2.5 and 7.4 and 12 and 37 pg, respectively. In Paltsev et al. (2013), the LOD for the DIM in rat plasma was set to 30 ng/ml. The authors used the HPLC-UV method. ...
... The intra-and inter-day performances of the assay were determined Also, DIM (20 mg/kg) was orally administered to rabbits (n = 3). (Reed et al., 2006), 4,4 0 -dichloro-DIM (Sepkovic et al., 2001) and 2 H-labeled DIM (Fujioka et al., 2014) were used as the IS or there was no IS at all (Paltsev et al., 2013). ...
... In the currently available literature, the authors used methyl tertbutyl ether to extract DIM from human and animal biological fluids (the proteins were previously precipitated with ice-cold ACN; Anderton et al., 2003;Fujioka et al., 2014;Hauder et al., 2011;Moussata et al., 2014) as well as diethyl ether (Reed et al., 2006), ethyl acetate (Paltsev et al., 2013) and chloroform (Sepkovic et al., 2001). It is noteworthy that the sample preparation methods described in these papers were labor-intensive and required a large consumption of extractant. ...
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Indole‐3‐carbinol is the subject of ongoing biomedical research due to its potential antiatherogenic, anticarcinogenic, and antioxidant effects. The antitumor properties are mainly associated with its major metabolite, i.e. 3,3′‐diindolylmethane (DIM). Typically, the biological activity of the chemical compound is manifested in the ng/mL concentration range. Consequently, the development of highly sensitive analytical methods to determine the DIM in various biological samples is an urgent issue. In this study, an HPLC–MS/MS method was established for the DIM quantification in human plasma. The developed method was validated according to the EMA guidelines. Sensitivity, selectivity, accuracy, and precision were found good allowing the DIM quantification in the concentration range of 5–500 ng/mL. The limit of detection and the lower limit of quantification were 1 and 5 ng/mL, respectively. 4‐Methoxy‐1‐methylindole was used as an internal standard (IS). The analytes were taken from the human plasma by the acetonitrile‐induced protein precipitation method with the addition of 3 mol/L ammonium sulfate as a salting‐out agent, which is a facile and efficient approach for high‐throughput bioanalysis. The chromatographic separation was performed on the Synergi Fusion‐RP C18 column (50 × 2.0 mm, 4 μm, 80 Å) under isocratic elution at 40°C. The mobile phase consisting of acetonitrile and water (0.1% formic acid) (85:15, v/v) was delivered at a flow rate of 0.20 mL/min. The DIM and IS were eluted at 2.36 ± 0.04 and 2.43 ± 0.03 min, respectively. The total analysis time was 3.20 min. Atmospheric pressure chemical ionization was carried out using multiple reaction monitoring in the positive polarity mode. The ion transitions were set to m/z 247.1 → 130.1 (DIM) and 162.1 → 147.1 (IS). The method was successfully applied to the analysis of plasma samples after a single oral administration of the Indinol® Forto drug (200 mg) to Russian female healthy volunteers. Also, the developed method was used for the analysis of rabbit plasma samples after a single oral dose of DIM (20 mg/kg).
... Co-incubation with isoform-selective CYP inhibitors suggested that CYP1A2 played the primary role in metabolism of DIM in MCF-7 cells. Surprisingly, in vivo pharmacokinetic studies in both mice, rats and humans have failed to report the formation of any phase 1 or phase 2 metabolite in plasma or urine (52, 103,106,108,109,[116][117][118][119]. In a recently published study with humans taking BioResponse DIM R , a formulation with clinically demonstrated enhanced absorption, we found extensive and rapid appearance of metabolites in both plasma (Figure 2) and urine (Figure 3) (20). ...
... Interestingly, 3-methylenehydroxy-DIM was not stable and spontaneously rearranged to a putative pyrano metabolite, 5a,6a,11b-tetrahydro-5-H-pyrano [,2,3-b:6,5-b'] diindole (pyrano-DIM). As has been observed in previous pharmacokinetic studies with I3C or DIM in humans (52, 103,106,108,109,[116][117][118][119], there was significant inter-individual variability (Figure 2). In MCF-7 cells the major mono-hydroxylated metabolite of DIM, 2-ox-DIM, failed to demonstrate any estrogenic activity. ...
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Hydrolysis of glucobrassicin by plant or bacterial myrosinase produces multiple indoles predominantly indole-3-carbinol (I3C). I3C and its major in vivo product, 3,3'-diindolylmethane (DIM), are effective cancer chemopreventive agents in pre-clinical models and show promise in clinical trials. The pharmacokinetics/pharmacodynamics of DIM have been studied in both rodents and humans and urinary DIM is a proposed biomarker of dietary intake of cruciferous vegetables. Recent clinical studies at Oregon State University show surprisingly robust metabolism of DIM in vivo with mono- and di-hydroxylation followed by conjugation with sulfate or glucuronic acid. DIM has multiple mechanisms of action, the most well-characterized is modulation of aryl hydrocarbon receptor (AHR) signaling. In rainbow trout dose-dependent cancer chemoprevention by dietary I3C is achieved when given prior to or concurrent with aflatoxin B1, polycyclic aromatic hydrocarbons, nitrosamines or direct acting carcinogens such as N-methyl-N'-nitro-nitrosoguanidine. Feeding pregnant mice I3C inhibits transplacental carcinogenesis. In humans much of the focus has been on chemoprevention of breast and prostate cancer. Alteration of cytochrome P450-dependent estrogen metabolism is hypothesized to be an important driver of DIM-dependent breast cancer prevention. The few studies done to date comparing glucobrassicin-rich crucifers such as Brussels sprouts with I3C/DIM supplements have shown the greater impact of the latter is due to dose. Daily ingestion of kg quantities of Brussels sprouts is required to produce in vivo levels of DIM achievable by supplementation. In clinical trials these supplement doses have elicited few if any adverse effects. Sulforaphane from glucoraphanin can act synergistically with glucobrassicin-derived DIM and this may lead to opportunities for combinatorial approaches (supplement and food-based) in the clinic.
... We performed UPLC-MS/MS analysis of plasma and urine prior to supplementation (T: À7 days), the morning (overnight fast) of [ 14 C]BaP microdosing (T: 0), and at subsequent timed intervals (plasma, 0.25, 0.50, 1.0, 1.5, 2.0, 3.0, 4.0, 8.0, 24, and 48 hours; urine 0-6, 6-12, 12-24, and 24-48 hours). In contrast to previously published studies (Sepkovic et al., 2001;Anderton et al., 2004;Reed et al., 2006Reed et al., , 2008Paltsev et al., 2013), we found rapid and significant sulfate and glucuronide conjugates of DIM in plasma and urine characterized by a marked interindividual variability. These conjugates were derived from 3-methylenehydroxy-DIM (tentatively identified as the breakdown product pyrano-DIM) with lesser amounts of 2ox-DIM and 3-methylenehydroxy-2-ox-DIM. ...
... The mean C max for DIM and the monohydroxylated metabolites and their respective conjugates are 423 ± 610 and 1910 ± 3190 pmol/ml plasma, respectively (Table 3). The T max and C max (average $0.4 mM) for DIM is consistent with previous studies (Reed et al., 2008;Paltsev et al., 2013). Mean noncompartmental half-life, apparent clearance, and apparent volume of distribution for DIM were 4.29 ± 2.48 hours, 161 ± 132 L x hr À1 , and 1010 ± 693 L, respectively. ...
Article
3,3'-Diindolylmethane (DIM), a major phytochemical derived from ingestion of cruciferous vegetables, is also a dietary supplement. In preclinical models, DIM is an effective cancer chemopreventive agent and has been studied in a number of clinical trials. Previous pharmacokinetic studies in preclinical and clinical models have not reported DIM metabolites in plasma or urine following oral dosing and the pharmacological actions of DIM on target tissues is assumed to be solely via the parent compound. Seven subjects (6 males and 1 female) ranging from 26-65 years of age, on a cruciferous vegetable-restricted diet prior to and during the study, took 2 BioResponse-DIM ® 150 mg capsules (45.3 mg DIM/capsule) every evening for one week with a final dose the morning of the first blood draw. A complete time course was performed with plasma and urine collected over 48 hours and analyzed by UPLC-MS/MS. In addition to parent DIM, two mono-hydroxylated metabolites and 1 di-hydroxylated metabolite, along with their sulfate and glucuronide conjugates, were present in both plasma and urine. Results reported here are indicative of significant phase 1 and phase 2 metabolism and differ from previous pharmacokinetic studies in rodents and humans which reported only parent DIM present following oral administration. 2-Ox-DIM, identified as one of the mono-hydroxylated products, exhibited greater potency and efficacy as an AHR agonist when tested in an XRE-luciferase reporter assay using Hepa1 cells. In addition to competitive phytochemical-drug adverse reactions, additional metabolites may exhibit pharmacological activity highlighting the importance of further characterization of DIM metabolism in humans. Significance Statement 3,3'-Diindolylmethane (DIM), derived from indole-3-carbinol in cruciferous vegetables, is an effective cancer chemopreventive agent in pre-clinical models and a popular dietary supplement currently in clinical trials. Pharmacokinetic studies to date found little or no metabolites in plasma or urine. In marked contrast, we demonstrate rapid appearance of mono- and di-hydroxylated metabolites in human plasma and urine as well as sulfate and glucuronide conjugates. 2-Ox-DIM exhibited significant AHR agonist activity emphasizing the need for characterization of pharmacological properties of DIM metabolites.
... Previously Heath et al. reported DIM peak plasma concentration to be 1 mM [79]. However, the administration of liquid DIM formulation with better bioavailability resulted in a 10-fold increase in С max [80]. Thus 10 mM was selected as potentially maximum tolerable clinical concentration. ...
Article
Epigenetic alterations represent promising therapeutic targets in cancer treatment. Recently it was revealed that small molecules have the potential to act as microRNA silencers. Capacity to bind the discrete stem-looped structure of pre-miR-21 and prevent its maturation opens opportunities to utilize such compounds for the prevention of initiation, progression, and chemoresistance of cancer. Molecular simulations performed earlier identified 3,3′-diindolylmethane (DIM) as a potent microRNA-21 antagonist. However, data on DIM and microRNA-21 interplay is controversial, which may be caused by the limitations of the cell lines.
... The natural indole glucosinolates decompose into I3C and form DIM in the stomach upon consumption. However, the bioavailability of DIM is poor and formulations are often necessary for in vivo tests [11,16] . Synthetic derivatives of DIM have been prepared by various synthetic methods [17,18] . ...
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Indoles of cruciferous vegetables are promising anti-tumor agents. Studies with indole-3-carbinol and its dimeric product, 3,3’-diindolylmethane (DIM), suggest that these compounds have the ability to deregulate multiple cellular signaling pathways that are essential for tumor growth and spread. These natural compounds are also effective modulators of transcription factors and non-coding RNAs. These effects explain their ability to inhibit tumor spread and to overcome drug resistance. In this work, pertinent literature on the effects of DIM and its synthetic derivatives on resistant tumors and resistance mechanisms in tumors is highlighted.
... The anti-cancer effects of DIM coincide with a broad spectrum of biological activities (Paltsev et al. 2013). In vitro studies demonstrate that DIM suppresses cancer cell proliferation, migration, and metastasis, and instead induces apoptosis in some cancer cell types (Azmi et al. 2008, Heo et al. 2018, Li et al. 2015. ...
Article
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The molecule 3,3'-diindolylmethane (DIM) is small, a major bioactive metabolite of indole-3 carbinol (13C), and a phytochemical compound from cruciferous vegetables released upon exposure to the gut acid environment. DIM is a proposed anti-cancer agent and was previously demonstrated to prevent radiation damage in the bone marrow and the gastrointestinal tract. Here we investigated the effect of DIM on radiation-induced injury to the lung in a murine model through untargeted metabolomics and gene expression studies of select genes. CBA mice were exposed to thoracic irradiation (17.5 Gy). Mice were treated with vehicle or DIM (250 mg kg, subcutaneous injection) on days -1 pre-irradiation through +14 post-irradiation. DIM induced a significant improvement in survival by day 150 post-irradiation. Fibrosis-related gene expression and metabolomics were examined using lung tissue from days 15, 45, 60, 90, and 120 post-irradiation. Our qRT-PCR experiments showed that DIM treatment reduced radiation-induced late expression of collagen Iα and the cell cycle checkpoint proteins p21/waf1 (CDKN1A) and p16ink (CDKN2A). Metabolomic studies of lung tissue demonstrated a significant dampening of radiation-induced changes following DIM treatment. Metabolites associated with pro-inflammatory responses and increased oxidative stress, such as fatty acids, were suppressed by DIM treatment compared to irradiated samples. Together these data suggest that DIM reduces radiation-induced sequelae in the lung.
... Met was used at 40 μM because this is near peak blood levels and near the dose previously tested by the Falcone lab [37]. BR-DIM was used at 20 μM because this dose slowed growth or killed cultured prostate cancer cells [26], and a peak of~7 μM BR-DIM was reported during in vivo exposure to mice [38]. Asa (salicylate) was used at 10 μM because this is a standard plasma level after enteric pill [39] and was the dose used in our previous study [3]. ...
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Purpose: This study tests whether metformin or diet supplement BR-DIM-induced AMP-activated protein kinase (AMPK) mediated effects on development are more pronounced in blastocysts or 2-cell mouse embryos. Methods: Culture mouse zygotes to two-cell embryos and test effects after 0.5-1 h AMPK agonists' (e.g., Met, BR-DIM) exposure on AMPK-dependent ACCser79P phosphorylation and/or Oct4 by immunofluorescence. Culture morulae to blastocysts and test for increased ACCser79P, decreased Oct4 and for AMPK dependence by coculture with AMPK inhibitor compound C (CC). Test whether Met or BR-DIM decrease growth rates of morulae cultured to blastocyst by counting cells. Result(s): Aspirin, metformin, and hyperosmotic sorbitol increased pACC ser79P ~ 20-fold, and BR-DIM caused a ~ 30-fold increase over two-cell embryos cultured for 1 h in KSOMaa but only 3- to 6-fold increase in blastocysts. We previously showed that these stimuli decreased Oct4 40-85% in two-cell embryos that was ~ 60-90% reversible by coculture with AMPK inhibitor CC. However, Oct4 decreased only 30-50% in blastocysts, although reversibility of loss by CC was similar at both embryo stages. Met and BR-DIM previously caused a near-complete cell proliferation arrest in two-cell embryos and here Met caused lower CC-reversible growth decrease and AMPK-independent BR-DIM-induced blastocyst growth decrease. Conclusion: Inducing drug or diet supplements decreased anabolism, growth, and stemness have a greater impact on AMPK-dependent processes in two-cell embryos compared to blastocysts.
... DIM has been reported to inhibit proliferation and induce programmed cell death in many types of cancer cells including breast, endometrium, colon, and prostate [9][10][11][12]. However, low oral bioavailability and hydrophobicity of DIM limit its passage through the tight junctions of the brain, thereby displaying poor anticancerous effects against high grade brain cancers [13][14][15][16]. In the present study we developed PLGA nanoencapsulated DIM with an aim to attain the following advantages: (i) to surmount the drug solubility problem, (ii) to enhance passaging of drugs selectively across the BBB, (iii) sustained drug release and (iv) to achieve high biocompatibility and tumor targeting efficiency. ...
Article
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GTSE1 over-expression has been reported as a potential marker for metastasis in various types of malignancies, including breast cancer. Despite this, the transcriptional regulation of this protein and the causes of its misregulation in tumors remain largely unknown. The aims of this work were to elucidate how GTSE1 is regulated at the transcriptional level and to clarify the mechanism underlying GTSE1-dependent cell functions in triple-negative breast cancer (TNBC).Here, we identified GTSE1 as a novel target gene of the TEAD4 transcription factor, highlighting a role for the YAP and TAZ coactivators in the transcriptional regulation of GTSE1.Moreover, we found that TEAD4 controls the formation of cell protrusions required for cell migration through GTSE1, unveiling a relevant effector role for this protein in the TEAD-dependent cellular functions and confirming TEAD4 role in promoting invasion and metastasis in breast cancer.Finally, we highlighted a role for the pRb-E2F1 pathway in the control of GTSE1 transcription and observed that treatment with drugs targeting the pRb-E2F1 or YAP/TAZ-TEAD pathways dramatically downregulated the expression levels of GTSE1 and of other genes involved in the formation of metastasis, suggesting their potential use in the treatment of TNBC.
... DIM has been reported to inhibit proliferation and induce programmed cell death in many types of cancer cells including breast, endometrium, colon, and prostate [9][10][11][12]. However, low oral bioavailability and hydrophobicity of DIM limit its passage through the tight junctions of the brain, thereby displaying poor anticancerous effects against high grade brain cancers [13][14][15][16]. In the present study we developed PLGA nanoencapsulated DIM with an aim to attain the following advantages: (i) to surmount the drug solubility problem, (ii) to enhance passaging of drugs selectively across the BBB, (iii) sustained drug release and (iv) to achieve high biocompatibility and tumor targeting efficiency. ...
Article
Current therapy for Glioblastoma is insufficient because of the presence of blood brain barrier. It limits the transport of essential drugs to the tumor sites. To overcome this limitation we strategized the delivery of an anticancer compound 3,3'-diindolylmethane by encapsulation in poly (lactic-co-glycolic acid) nanoparticles. These nanoparticles were tagged with a novel peptide against somatostatin receptor 2 (SSTR2), a potential target in glioma. The nanoformulation (27-87nm) had loading and encapsulation efficiency of 7.2% and 70% respectively. It was successfully internalized inside the glioma cells resulting in apoptosis. Furthermore, an in vivo bio-distribution study revealed the selective accumulation of the nanoformulation into rat brain tumor sites by crossing the blood brain barrier. This resulted in abrogation of epidermal growth factor receptor pathway activation in glioma cells. Our novel nanopreparation therefore shows great promise to serve as a template for targeted delivery of other therapeutics in treating GBM.