Dibromoacetic acid   中文名称: 二溴乙酸

当暴露于 DBAA (5–40 μM) 时，体外胸腺细胞增殖显着降低[1]。 DBAA 处理 (5–40 μM) 诱导细胞周期停滞 24 小时。根据数据[1]，用不同浓度的 DBAA 处理的胸腺细胞在 G0 /G1 期至少增加 40%，在 S 期减少 50%。在 24 小时内，DBAA (5–40 μM) 会增加 Fas/FasL 的表达并减少 Bcl-2 的表达[1]。

基于雄性大鼠恶性间皮瘤的发生率较高，有一些迹象表明二溴乙酸具有致癌潜力。在雄性大鼠中观察到的单核细胞白血病发病率升高可能与接触二溴乙酸有关[2]。雌性大鼠中的二溴乙酸活性是基于单核细胞白血病的阳性趋势和较高的频率[2]。根据肝细胞肿瘤和肝母细胞瘤（仅限雄性）发生率的升高，二溴乙酸对雄性和雌性小鼠均具有明显的致癌作用。雄性小鼠的肺部肿瘤发生率也较高，这被认为与暴露有关[2]。

细胞增殖测定[1]
细胞类型： BALB/c 小鼠的胸腺细胞
测试浓度： 0、5、10、20 和 40 μM
孵育持续时间：6、12、24、48 和 72 小时
实验结果：导致细胞对 T- 的增殖反应显着减弱细胞有丝分裂原6小时或更长时间。 6 小时时，仅在 40 μM 浓度下观察到显着抑制，而在 24、48 和 72 小时时，所有浓度均观察到显着抑制。

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蛋白质印迹分析[1]
细胞类型：胸腺细胞
测试浓度：0、5、10、20 和 40 μM
孵育时间：24小时
实验结果：Fas/FasL的表达量从10 μM开始急剧增加，Bcl-2的表达量增加所有浓度均减弱。

Animal/Disease Models: Male and female F344/N rats and B6C3F1 mice[2]
Doses: Groups of five male and five female rats/mice were exposed to 0, 125, 250, 500, 1,000, or 2,000 mg/L Dibromoacetic acid in drinking water for 2 weeks Groups of 10 male and 10 female rats /mice were exposed to 0, 125, 250, 500, 1,000, or 2,000 mg/L Dibromoacetic acid in drinking water for 3 months Groups of 50 male and 50 female rats/mice were exposed to drinking water containing 0, 50, 500, and 1,000 mg/L Dibromoacetic acid for 2 years
Route of Administration: Exposed to Dibromoacetic acid (greater than 99% pure) in drinking water for 2 weeks, 3 months, or 2 years.
Experimental Results: Exposure to Dibromoacetic acid for 2 years caused increased incidences of cystic degeneration of the liver in male rats, increased incidences of alveolar epithelial hyperplasia and nephropathy in female rats, and increased incidences of splenic hematopoiesis in male mice.

Absorption, Distribution and Excretion
In the preliminary reproductive/developmental toxicity studies (rats), dibromoacetic acid (DBA) was added to drinking water. The studies included the absorption and biodistribution of DBA, including its entry into the placenta, amniotic fluid, fetus, or breast milk. Each group in the preliminary reproductive/developmental toxicity studies of DBA consisted of 50 Sprague-Dawley rats (per sex per group). DBA (0, 125, 250, 500, or 1000 ppm) was added to drinking water from 14 days prior to mating until gestation and lactation (63 to 70 days). …Satellite groups (6 male rats and 17 female rats per group per study)…were used for bioanalytical sampling. Rats…drank less water due to a marked taste aversion to DBA, especially in parent animals at the two highest exposure levels (500 and 1000 ppm DBA). DBA intake (mg/kg/day) was slightly higher in female rats than in male rats, especially during pregnancy and lactation; the highest intake (mg/kg/day) was observed in weaned rats. DBA reached detectable and quantifiable concentrations in plasma, placenta, amniotic fluid, and milk. Plasma samples confirmed that rats primarily drank water in the dark; this drinking pattern, rather than accumulation, resulted in sustained DBA concentrations in plasma for 18 to 24 hours…
DBA concentrations were determined in the testicular interstitial fluid of male Sprague-Dawley rats that received 250 mg/kg body weight of DBA via gavage for five consecutive days. Body weight… The concentration of dibromoacetic acid in the testicular fluid peaked at 79 μg/mL (approximately 370 μM) 30 minutes after the last administration, with a half-life of approximately 1.5 hours.
Dibromoacetic acid (DIA) was added to the drinking water of Sprague-Dawley rats at concentrations ranging from 125 to 1000 ppm (mg/L), starting 14 days before cohabitation and continuing into pregnancy and lactation… Quantifiable amounts of DIA were detected in parental and fetal plasma, placental tissue, amniotic fluid, and breast milk. Therefore, DIA can cross the placenta and be absorbed by fetal tissues.
The oral bioavailability of DIA in male F344/N rats has been reported to be 30%… Compared to dichloroacetic acid, the lower bioavailability of DIA is due to its… first-pass metabolism in the liver.
Biological half-life

The DIA content in the testicular interstitial fluid of male Sprague-Dawley rats administered 250 mg/kg body weight of DIA by gavage for five consecutive days was measured… The half-life was approximately 1.5 hours.

Interactions
Studies have found that the disinfection byproduct dibromoacetic acid (DBA) increases the concentrations of circulating estradiol (E2) and estrone (E1) in female rats. This effect is clearly at least partly due to the inhibition of hepatic catabolism. This study aimed to investigate whether DBA could enhance hypothalamic upregulation to trigger a luteinizing hormone (LH) surge by increasing sex hormone levels, or affect the ability of the neurotoxin sodium dimethyl dithiocarbamate (DMDC) to block LH surges. Sprague-Dawley rats were administered DBA (0–150 mg/kg) by gavage for 14 consecutive days, and ovariectomy was performed on day 11, with estradiol capsules implanted to induce daily LH surges. 0.1 mM/kg DMDC was injected at 13:00 on day 14, and blood samples were collected that afternoon. DBA induced a dose-dependent increase in total estrogen levels. For the identified LH peak, the area under the LH curve was divided into two groups, corresponding to two low-dose groups (0 and 37.5 mg/kg DBA) and two high-dose groups (75 and 150 mg/kg DBA). Therefore, comparisons were made between the low-dose and high-dose groups, revealing significant differences between them. In the 150 mg DBA/0.1 mM DMDC group, the time to identifiable LH peak was comparable to that of female mice not treated with DMDC, while the time to peak was delayed in the 37.5 mg DBA/0.1 mM DMDC group. No significant effect was observed with DBA treatment alone. These results indicate that exposure to DBA leads to a dose-dependent increase in total estrogen concentration, while the blocking effect of DMDC on the LH peak is weakened. This effect appears to be attributed to enhanced upregulation of estrogen-related brain mechanisms, thereby stimulating LH peak production. Chlorination of drinking water produces disinfection byproducts (DBPs), and studies have shown that high doses of DBPs disrupt spermatogenesis in rodents, suggesting that DBPs may pose a risk to male reproduction. ...A cohort study aimed to assess semen quality in men with defined DBP exposure. ...The results of this study do not support an association between DBP exposure levels near regulatory limits and poor sperm outcomes, although an association was found between total organohalides and sperm concentration. ...The only association between total organohalides exposure and sperm concentration may support the following findings: total organohalides are more likely to cause poor pregnancy outcomes than any regulated disinfection byproduct (DBP) class or type, and the toxicity of total organohalides is greater than that of individual or subclasses of disinfection byproducts. .../Disinfection Byproducts/

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Non-human Toxicity Values
Oral LD50 in rats: 1737 mg/kg

[1]. Shu-Ying Gao, et al. Dibromoacetic Acid Induces Thymocyte Apoptosis by Blocking Cell Cycle Progression, Increasing Intracellular Calcium, and the Fas/FasL Pathway in Vitro.Toxicol Pathol. 2016 Jan;44(1):88-97.
[2]. National Toxicology Program. Toxicology and carcinogenesis studies of dibromoacetic acid (Cas No. 631-64-1) in F344/N rats and B6C3F1 mice (drinking water studies). Natl Toxicol Program Tech Rep Ser. 2007 Apr;(537):1-320.

According to data from the National Toxicology Program (NTP), dibromoacetic acid (DBA) may be carcinogenic. DBA is a monocarboxylic acid formed when two methyl hydrogen atoms in the acetic acid molecule are replaced by bromine atoms. It is a marine metabolite that induces apoptosis and delays aging. It is a monocarboxylic acid and also a 2-bromocarboxylic acid. Its function is related to acetic acid. DBA has been reported in Asparagopsis taxiformis, and relevant data are available. Mechanism of Action…The ability of dibromoacetic acid (DBA) to induce DNA hypomethylation, glycogen accumulation, and peroxisome proliferation was investigated… Female B6C3F1 mice and male Fischer 344 rats were given DBA solutions at concentrations of 0, 1000, and 2000 mg/L, respectively. Animals were euthanized after 2, 4, 7, and 28 days of exposure. DBA resulted in a dose- and time-dependent decrease in the content of 5-methylcytosine in DNA of 20% to 46%. Hypomethylation of the c-myc gene was observed in mice after 7 days of exposure to dibromoacetic acid (DBA). The methylation level of 24 CpG sites in the insulin-like growth factor 2 (IGF-II) gene decreased from 80.2% ± 9.2% to 18.8% ± 12.9% after 28 days of treatment with 2000 mg/L DBA. Dibromoacetic acid increased the mRNA expression of both c-myc and IGF-II genes in mouse liver. A dose-dependent increase in c-myc mRNA expression was also observed in rats. In both mice and rats, DBA induced dose-dependent glycogen accumulation and increased peroxisome lauroyl-CoA oxidase activity. Therefore, similar to dichloroacetic acid and trichloroacetic acid, DBA induces hypomethylation of DNA and both c-myc and IGF-II genes, increases the mRNA expression of these two genes, and leads to peroxisome proliferation. Similarly, DBA also induces glycogen accumulation. These results indicate that DBA shares common biochemical and molecular activities with dichloroacetic acid and/or trichloroacetic acid, suggesting that it may also be a hepatotoxic carcinogen. Haloacetic acids (HA) are common embryotoxic contaminants in drinking water. The embryotoxic mechanism of HA may be partly mediated by inhibition of protein kinase C (PKC). This study aimed to evaluate the pathogenesis of hyaluronic acid (HA) embryotoxicity and compare these data with those of specific (Bis I) and non-specific (astrosporin) protein kinase C (PKC) inhibitors. Embryos were incubated with various HA, Bis I, asteroidin, or Bis V (negative control) for different durations. Cell cycle analysis was performed by flow cytometry after propidium iodide (PI) staining; apoptosis was assessed by fluorescence microscopy after LysoTracker staining. At concentrations producing 100% embryotoxicity but not lethality, only asteroidin disrupted the cell cycle. However, flow cytometry analysis showed that sub-G1 phase events (an apoptosis marker) accumulated over time after treatment with bromochloroacetic acid, dichloroacetic acid, and astrococcus, while this phenomenon was not observed after treatment with dibromoacetic acid, Bis I, or Bis V. Sub-G1 phase events were particularly pronounced in the head region, while in the heart they remained at control levels. Lysosomal tracer staining confirmed a similar apoptotic pattern in intact embryos; BCA and DCA produced strong staining in the forebrain, but almost no staining in the heart. These data suggest that while cell cycle dysregulation may not be the pathogenic mechanism of HA embryotoxicity, these drugs do induce embryonic cell apoptosis. Furthermore, the lack of apoptosis induced by Bis I suggests that PKC inhibition is unlikely to be the sole mediator of HA embryotoxicity. The relevant mechanisms of HA carcinogenicity include the oncogenic mechanisms of DCA and TCA. Clearly, there is more than one mechanism leading to the effects of these compounds, and these mechanisms vary in importance to the activity of different members of this class of compounds. Some of these mechanistic differences may be related to differences in the induced tumor phenotypes. One phenotype appears to be associated with previous characterizations of peroxisome proliferator-induced tumors and is induced by trichloroacetic acid (TCA). A second phenotype involves tumors with low glycogen content that show strong staining responses to c-Jun and c-Fos antibodies. This phenotype is produced by dichloroacetic acid (DCA). These effects may be due to the selective induction of lesions with different defects in cell signaling pathways that control cell division and cell death. Brominated heterocyclic amines (HAAs) are about 10 times more likely to induce point mutations than their chlorinated analogs. This does not necessarily demonstrate that they induce cancer in vivo through mutagenic mechanisms, but such activity must be considered as data on their carcinogenic activity become more comprehensive. Heterocyclic amines vary considerably in their ability to induce oxidative stress and increase the content of 8-hydroxydeoxyguanosine (8-OH-dG) in liver nuclear DNA. This characteristic is particularly pronounced in brominated compounds. Notably, brominated analogs are not more likely to induce liver tumors than their corresponding chlorinated heterocyclic amines. Therefore, it remains questionable whether this mechanism is the most important factor determining this effect.

C2H2BR2O2

217.84

215.842

631-64-1

12433

Hygroscopic crystals

2.382 g/mL at 25ºC(lit.)

128-130ºC16 mm Hg(lit.)

32-38ºC(lit.)

>230 °F

1.598

1.186

37.3

1

2

1

6

60.6

0

C(C(=O)O)(Br)Br

SIEILFNCEFEENQ-UHFFFAOYSA-N

InChI=1S/C2H2Br2O2/c3-1(4)2(5)6/h1H,(H,5,6)

2,2-dibromoacetic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : 100 mg/mL (459.05 mM)

配方 1 中的溶解度: 2.5 mg/mL (11.48 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液; 超声助溶。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 2 中的溶解度: ≥ 2.5 mg/mL (11.48 mM) (饱和度未知) in 10% DMSO + 90% Corn Oil (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL 澄清 DMSO 储备液添加到 900 μL 玉米油中并混合均匀。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
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10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
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