In HepG2 and C2C12 cells, dantrofrine (0.1, 1, and 10 μM) dose-dependently increased AMPK and acetyl-CoA carboxylase (ACC) phosphorylation. Simultaneously, the administration of Danthron resulted in a considerable decrease in the expression of the fatty acid synthase (FAS) and sterol regulatory element binding protein 1c (SREBP1c) genes, as well as in the levels of total cholesterol (TC) and triglycerides (TG). Furthermore, the use of Danthron effectively raises glucose consumption. Danthron activates the AMPK signaling system, which efficiently lowers intracellular lipid content and promotes in vitro glucose intake. For HepG2 cells, 10 μM Danthron/24 hours is safe. HepG2 cells were cultured with Danthron (0.1–10 μM) in FBS-free media for 8 hours after they reached 80% confluence. Cells were then extracted in preparation for the Western blot test. While there are no changes in t-AMPK protein, there is a dose-dependent rise in p-AMPK protein while using danthron [1]. With an IC50 of 0.11 μM, danthron inhibits the transactivation of retinoic acid X receptor alpha (RXRα) produced by 9-cis retinoic acid (9cRA). Using isothermal titration calorimetry (ITC) assays, the stoichiometry of Danthron binding to the RXRα-ligand binding domain (LBD) was further clarified. The ITC experiment yielded a KD value of 7.5 μM for Danthron binding to RXRα-LBD[2].

Danthron functions as an insulin sensitizer in the body. Danthron enhances insulin sensitivity in mice with diet-induced obesity (DIO). Insulin tolerance test results demonstrated that Danthron (5 mg/kg)-treated diet-induced obese mice displayed lower blood glucose levels after insulin challenge compared with the control group [2].

Absorption, Distribution and Excretion
Following its administration within 24 hr of the induction of labor in 12 women, dantron was found in maternal urine, neonatal urine and amniotic fluide. Most of the drug appeared as a glucuronide in both mothers and babies.
Male Wistar rats were given the sodium salt of dantron intravenously at 4.8, 22 or 58 umol/kg (1.2, 5.3 or 14 mg/kg) bw or at 12 umol/kg (28.8 mg/kg) bw by gastric tube. ... Following intravenous administration, about 80% of the dantron conjugates in bile were excreted after 1 hr; the dose fractions found after 5 hr represented about 20%, 30% and 40% of the low-; intermediate- and high-dose levels, respectively. The corresponding fractions in urine were 16%, 12% and 10%, giving rise to bile:urine excretion ratios of 1.3, 2.7 and 4.0, respectively. Only 30-50% of the dose could be accounted for by conjugates. Earlier studies also showed that after oral administration of dantron only 30-40% of the total dose administered could be recovered in feces and urine, mostly during the first 24 hr. /Dantron sodium salt/
Like other anthraquinone compounds, dantron is partially absorbed from the small intestine.
Rats were infused with danthron (I) at doses of 0.48, 2.2 and 5.8 umol/100 g body weight, or given 12 umol/100 g with gastric tube. TLC of bile and urine demonstrated a number of metabolites, at both administration routes. These included danthron monosulfate (II) and -glucuronide (III), two other phase 2 metabolites which behaved as the corresponding diconjugates, and several phase 1 metabolites (IV) in conjugated form. ... Following infusion, about 80% of the danthron conjugates in bile were excreted after 1 hour; the dose fractions found after 5 hours represented about 20%, 30%, and 40% at the low, intermediate and high dose level, respectively. The corresponding fractions in urine were 16%, 12% and 10%, giving rise to bile:urine excretion ratios of 1.3, 2.7 and 4.0, respectively. This change in excretion pattern was associated with changes in metabolite muster, which involved a decrease in the balance of IV:I conjugates, as well as an increase in III:II ratio. IV was more abundantly present in bile than in urine, and showed a more sustained excretion than the danthron conjugates. By intragastric administration, the cumulated excretion (bile + urine) of I conjugates were only 6%, 8% and 5% of dose, in three consecutive 6 hours' periods (0-6, 6-12 and 12-18 hours after dosing). The bile:urine excretion ratios seemed to decrease with time, as did the III:II ratio...

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Metabolism / Metabolites
In vitro, rat jejunum and colon transformed dantron into its monoglucuronide and monosulfate, the monoglucuronide being the major metabolite.
Male Wistar rats were given the sodium salt of dantron intravenously at 4.8, 22 or 58 umol/kg (1.2, 5.3 or 14 mg/kg) bw or at 12 umol/kg (28.8 mg/kg) bw by gastric tube. Metabolites identified in the bile and urine following administration by either route included the monosulfate, beta-glucuronide and other unidentified metabolites. /Dantron sodium salt/
Danthron infused intravenously in rats shows a complex dose-dependent pattern of metabolism and excretion. The metabolites, particularly the more polar ones, are in general excreted predominantly in bile, to a lesser extent in urine. ... /This/ paper describes a further study within a bile-derived metabolite group, which proved to be particularly heterogeneous. It contained more than a dozen metabolites, which were conjugates of four different aglycons including the parent danthron...
... Everted sacs of rat jejunum and stripped colon were filled with Krebs-Henseleit solution (K-H) on the serosal (BL) side, and bathed at the mucosal (LU) side with K-H containing either danthron (3-4 nmol/mL) or rhein (10 nmol/mL). After 60 min incubation at 37 degrees C, LU and BL solutions and gut tissue were analysed for parent diphenol and metabolites by reverse-phase high-pressure liquid chromatography. Reference metabolites were isolated and purified from urine and bile of rats infused with danthron or rhein. The studies showed: (1) only small amounts of unchanged drug were present on the contraluminal side; (2) in both tissues, danthron was transformed into its monoglucuronide (G) and monosulfate (S); the ratio G:S was 6-8:1 in jejunum, and even greater in colon; (3) in jejunum, G and S were mainly secreted (LU:BL distribution ratios greater than 10:1); (4) in the colon, however, the main G fraction was absorbed (BL:LU ratios of 3:1), whereas a slight net secretion of S seemed to take place; (5) residuals (%) in gut tissue were small; (6) rhein was more slowly taken up and metabolized, but seemed otherwise to behave as danthron...

Interactions
The modifying effects of chrysazin on 1,2-dimethylhydrazine (DMH)-induced colon and liver carcinogenesis were examined in male ICR/CD-1 mice. Starting at 6 weeks of age, mice were divided into four groups, two of which were treated with sc injections of DMH (20 mg/kg body wt) once a week for 12 weeks. A week after the final injection of DMH, one group was kept on the basal diet throughout the study (group I), and the other group was fed the diet containing chrysazin (mixed in basal diet at 0.2% concentration) alone for 42 weeks (group II). The other two groups were injected with normal saline and given the diet containing 0.2% chrysazin for 42 weeks (group III), or the basal diet during the experiment (group IV). The incidence and multiplicity of colon tumors of group II were significantly greater than those of group I (P < 0.05, P < 0.01). The incidence and multiplicity of the hepatocellular neoplasms of group II were larger than those of group I (P < 0.002, P < 0.02 respectively). In group III, colon tumors were not found, though a few liver neoplasms and severe inflammatory lesions of the colon were observed. The activity of ornithine decarboxylase of the colonic mucosa in mice exposed to chrysazin was stronger than that of animals without chrysazin. The results suggest that the promoting effect of chrysazin is probably related to an increase of cell proliferation in the target organ. A synergistic effect of DMH with chrysazin was also observed in liver tumorigenesis.
When danthron was administered in the feed to mice that also received 1,2- dimethylhydrazine, the incidence and multiplicity of adenomas of the colon and liver were significantly increased.
The tumor-promoting activity of the anthraquinone laxative danthron was studied by giving 3 groups of male rats a single subcutaneous injection of the colon tumor-inducing agent 1,2-dimethylhydrazine (DMH). After 1 week, the animals were fed diets containing 0, 600 or 2400 ppm of danthron for 26 weeks. Two other groups of rats were included in the study; one received no treatment while the other was given danthron only. Altogether 9 tumors were observed among animals given DMA with or without danthron. The incidence of colon tumors was higher in animals receiving DMH and danthron than in those given DMH only (5/60 vs. 0/30), but this difference was not statistically significant. The kidneys and lymph nodes of mesocolon were enlarged and showed a yellowish-red and brown discoloration, respectively. The pigment mostly displayed a PAS-positive reaction but contained no lipid as determined by several staining procedures. The available evidence suggests that the pigment is drug-derived.

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Non-Human Toxicity Values
LD50 Mouse oral < 7 g/kg[The Merck Index, Fourteenth Edition (2006)
LD50 Mouse ip 500 mg/kg

[1]. Danthron activates AMP-activated protein kinase and regulates lipid and glucose metabolism in vitro. Acta Pharmacol Sin. 2013 Aug;34(8):1061-9.

[2]. Danthron functions as a retinoic X receptor antagonist by stabilizing tetramers of the receptor. J Biol Chem. 2011 Jan 21;286(3):1868-75.

Dantron (Chrysazin; 1,8-Dihydroxyanthraquinone) can cause cancer according to The World Health Organization's International Agency for Research on Cancer (IARC).
Danthron is an orange crystalline powder. Almost odorless and tasteless. (NTP, 1992)
Chrysazin is a dihydroxyanthraquinone that is anthracene-9,10-dione substituted by hydroxy groups at positions 1 and 8. It has a role as an apoptosis inducer and a plant metabolite.
Withdrawn from the Canadian, US, and UK markets in 1998 due to genotoxicity.
Danthron has been reported in Senna obtusifolia, Asphodelus fistulosus, and other organisms with data available.
Danthron is a reddish, synthetic anthraquinone derivative. Danthron has been widely used as a laxative, but is no longer used to treat constipation and is currently used as an antioxidant in synthetic lubricants, in the synthesis of experimental antitumor agents, as a fungicide and as an intermediate for making dyes. This substance is a suspected mutagen and is reasonably anticipated to be a human carcinogen based on evidence of carcinogenicity in experimental animals. (NCI05)

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Mechanism of Action
The mechanism of metal-mediated DNA damage by carcinogenic danthron (1,8-dihydroxyanthraquinone) and anthraquinone was investigated by the DNA sequencing technique using 32P-labeled human DNA fragments obtained from the human c-Ha-ras-1 protooncogene and the p53 tumor suppressor gene. Danthron caused DNA damage particularly at guanines in the 5'-GG-3', 5'-GGGG-3', 5'-GGGGG-3' sequences (damaged bases are underlined) in the presence of Cu(II), cytochrome P450 reductase and the NADPH-generating system. The DNA damage was inhibited by catalase and bathocuproine, suggesting the involvement of H2O2 and Cu(I). The formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine increased with increasing concentration of danthron. On the other hand, carcinogenic anthraquinone induced less oxidative DNA damage than danthron. Electron spin resonance study showed that the semiquinone radical could be produced by P450 reductase plus NADPH-mediated reduction of danthron, while little signal was observed with anthraquinone. These results suggest that danthron is much more likely to be reduced by P450 reductase and generate reactive oxygen species through the redox cycle, leading to more extensive Cu(II)-mediated DNA damage than anthraquinone. In the case of anthraquinone, its hydroxylated metabolites with similar reactivity to danthron may participate in DNA damage in vivo. /It was concluded/ that oxidative DNA damage by danthron and anthraquinone seems to be relevant for the expression of their carcinogenicity.
... All three tested anthraquinones, emodin, aloe-emodin, and danthron, showed capabilities to inhibit the non-covalent binding of bisbenzimide Hoechst 33342 to isolated DNA and in mouse lymphoma L5178Y cells comparable to the topoisomerase II inhibitor and intercalator m-amsacrine. In a cell-free decatenation assay, emodin exerted a stronger, danthron a similar and aloe-emodin a weaker inhibition of topoisomerase II activity than m-amsacrine. Analysis of the chromosomal extent of DNA damage induced by these anthraquinones was performed in mouse lymphoma L5178Y cells. Anthraquinone-induced mutant cell clones showed similar chromosomal lesions when compared to the topoisomerase II inhibitors etoposide and m-amsacrine, but were different from mutants induced by the DNA alkylator ethyl methanesulfonate. These data support the idea that inhibition of the catalytic activity of topoisomerase II contributes to anthraquinone-induced genotoxicity and mutagenicity.

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Therapeutic Uses
Danthron has been widely used since the beginning of this century as a laxative. In 1987, the FDA ordered its withdrawal from the market for its use as a laxative, and U.S. manufacturers voluntarily withdrew production of all human drug products containing the compound. /Former use in US/
Therapeutic Indications: Constipation in terminally ill patients.

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Drug Warnings
Contraindications: In common with other gastro-intestinal evacuants, Co-danthramer capsules should not be given when acute or painful conditions of the abdomen are present or when the cause of the constipation is thought to be an intestinal obstruction. Hypersensitivity to any of the constituents of the product. Peanut or soya allergies.
Dantron may cause temporary harmless pink or red coloring of the urine and peri-anal skin. With prolonged high dosage the mucosa of the large intestine may become colored.
Co-danthramer capsules are contraindicated in pregnant women and nursing mothers.
A woman developed deep discoloration of the skin following ingestion of large amounts of a laxative containing dantron. Such staining was also found in other studies, predominantly in elderly subjects, and was localized to the buttocks and thighs, with minor inflammatory symptoms. Contact of skin with feces or urine containing the drug seems to be a prerequisite for discoloration. Inflammation, when present, may result from reduction of the parent compound in the colon to the diol derivative, which irritates both the gut and skin, while the parent compound does not.
For more Drug Warnings (Complete) data for 1,8-Dihydroxyanthraquinone (6 total), please visit the HSDB record page.

C14H8O4

240.2109

240.042

117-10-2

2950

Brown to breen solid powder

1.5±0.1 g/cm3

452.7±35.0 °C at 760 mmHg

191-193 °C(lit.)

241.7±22.4 °C

0.0±1.1 mmHg at 25°C

1.733

4.57

2

4

0

18

346

0

QBPFLULOKWLNNW-UHFFFAOYSA-N

InChI=1S/C14H8O4/c15-9-5-1-3-7-11(9)14(18)12-8(13(7)17)4-2-6-10(12)16/h1-6,15-16H

1,8-dihydroxyanthracene-9,10-dione

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 : ~5 mg/mL (~20.82 mM)

Solubility in Formulation 1: 10 mg/mL (41.63 mM) in 50% PEG300 +50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication (<60°C).
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)