In MKN45 cells, α-amanitin lowers TAF15 mRNA and protein levels and prevents RNAPII from acting on TAF15 mRNA [2]. At doses of 100, 10, 1, 0.1, and 0.01 μg/mL, α-amanitin decreased cell viability by 14%, 21%, 41%, 44%, and 50%, respectively. At 36 hours, the α-Amanitin's LD50 was determined to be 1 μg/mL. At a concentration of 1 μg/mL, α-Amanitin considerably raised the total amount of intracellular protein at 24 hours in comparison to the control [3]. The cumulus cell gap junction genes (Gja1, Gja4, and Gjc1), as well as FSHr and LHr, are much less expressed when α-amanitin is present [4].

After intravenous administration, the α-Amanitin's LD50 in BALB/c mice was 0.327 mg/kg. Serum levels of WBC, RBC, and HGB dramatically decreased twelve hours after α-Amanitin was injected into the tail vein, whereas serum levels of BUN and Crea significantly increased. Certain genes (Hsp90b1, Irx4, etc.) are inhibited by α-amanitin, and the protein it encodes controls the activity of RNA polymerase II. Certain transcription-related proteins, including as Nmi and Trpc5, are down-regulated by α-amanitin [1]. α-Amanitin exhibits strong DTC inhibition properties. Whereas the body weight of mice receiving intraperitoneal injection of MKN45 cells continued to drop, the body weight of mice injected with cells treated with α-Amanitin (0.4 mg/kg, ip) remained stable [2].

Metabolism / Metabolites: Free toxin may be removed by opsonization via the reticuloendothelial system (primarily the liver and kidneys) or it may be degraded through cellular internalization via the lysosomes. Lysosomes are membrane-enclosed organelles that contain an array of digestive enzymes, including several proteases.

Effects During Pregnancy and Lactation
◉ Overview of Drug Use During Lactation
Amanita toxins are water-soluble, heat-resistant polypeptides found in species of the genera Amanita (most commonly Amanita phalloides), Galerina, and some Amanita. The main toxin in Amanita phalloides is α-amanitin, a cyclic octapeptide. It is a potent RNA polymerase inhibitor that blocks the production of mRNA and protein synthesis in liver and kidney cells. There was one case where an infant who breastfed 11.5 hours after its mother ingested Amanita phalloides developed elevated liver enzymes. However, two recent documented cases show that breastfed infants did not experience adverse reactions, and amanita toxins were not detected in breast milk. However, mothers suspected of Amanita phalloides poisoning should not breastfeed until they recover or breast milk toxicology screening rules out toxicity.
◉ Impact on Breastfed Infants
In Germany, a 20-year-old breastfeeding mother consumed a meal consisting entirely of mushrooms, reportedly Amanita phalloides. The following morning, approximately 11.5 hours after consuming the mushrooms, she fed her 10-week-old, 5-kilogram infant. This feeding consisted of 80-100 ml of breast milk and an equal amount of ready-to-feed infant formula (Milasan-Neu). At this point, the mother developed symptoms of poisoning (vomiting and diarrhea). Due to her worsening condition, she was unable to continue breastfeeding, and the baby was subsequently fed formula. The mother was hospitalized for Amanita phalloides poisoning, with aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT) levels of 10,000 and 40,000 respectively (normal values are approximately 500-550). The infant was admitted to a children's hospital for observation. No abnormalities were found in the infant's relatives, and clinical examination upon admission revealed no obvious signs of liver, brain, or blood system disease. Six days after the mother ingested the poisonous Amanita mushroom, the infant's laboratory test results (electrolytes, serum protein electrophoresis, bilirubin, gamma-glutamyl transferase, alkaline phosphatase, creatinine, blood glucose, urine tests, prothrombin time (PTT), and prothrombin time (PT) [rapid test]) were all normal, except that ASAT and ALAT values were approximately twice the normal range. These values slowly decreased and returned to normal approximately 40 days after ingestion. A 32-year-old mother shared a meal of freshly picked mushrooms (Amanita phalloides) with her family and developed symptoms 15 hours after ingestion. She went to the emergency department 29 hours after ingestion and was found to have significantly elevated liver enzymes. Her 4-month-old daughter was breastfed 4 hours after ingestion. The asymptomatic infant was evaluated 48 hours after breastfeeding and was discharged from the emergency department without signs of hepatotoxicity.
A 33-year-old woman picked approximately 200 mushrooms in a forest in France. She cooked and ate some, and 11 hours after ingestion, she experienced nausea, vomiting, and diarrhea. She was taken to the hospital for treatment and diagnosed with elevated liver enzymes. Within 36 hours of consuming the mushrooms, she breastfed her 5-month-old daughter three times a day. Her daughter was taken to the hospital but showed no symptoms or physiological disturbances.
◉ Effects on lactation and breast milk
As of the revision date, no relevant published information was found.

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Interactions
...It has been reported that certain drugs can protect mice from lethal doses of phalloidin and amatoxins (including muscarine). These drugs include high doses of penicillin, chloramphenicol, phenylbutazone, etc. However, these antidotes have not undergone sufficient clinical trials and are therefore not currently recommended for use. /Amatoxins/
Methylmercury stimulates RNA synthesis in the nucleus of HeLa cells. This stimulation specifically targets α-amaminine-sensitive RNA synthesis (catalyzed by RNA polymerase); conversely, α-amaminine-resistant synthesis (catalyzed by RNA polymerases I and III) is inhibited.

[1]. Pathological effects of the mushroom toxin alpha-amanitin on BALB/c mice. Peptides. 2006 Dec;27(12):3047-3052.

[2]. α-Amanitin Restrains Cancer Relapse from Drug-Tolerant Cell Subpopulations via TAF15. Sci Rep. 2016 May 16;6:25895.

[3]. Evaluation and comparison of alpha- and beta-amanitin toxicity on MCF-7 cell line. Turk J Med Sci. 2014;44(5):728-32.

[4]. RNA Polymerase II Inhibitor, α-Amanitin, Affects Gene Expression for Gap Junctions and Metabolic Capabilities of Cumulus Cells, but Not Oocyte, during in vitro Mouse Oocyte Maturation. Dev Reprod. 2013 Mar;17(1):63-72.

Alpha-amanin has been found in Amanita suballiacea, Amanita phalloides, and other organisms with relevant data. It is a cyclic octapeptide with cysteine and tryptophan linked by a thioether bridge. It inhibits RNA polymerase II. Poisoning may require a liver transplant. Mechanism of Action: The long-term, delayed hepatotoxicity observed in human poisoning is more likely due to α-, β-, and γ-amanin, especially the α-amanin component. These so-called amantoxins are more toxic than phalloidin and, unlike the latter, damage the nucleolus of hepatocytes, subsequently damaging the nucleus.

C₃₉H₅₄N₁₀O₁₄S

918.97

918.354

23109-05-9

441541

White to yellow solid powder

1.57 g/cm3

1622.2ºC at 760 mmHg

254-255ºC(lit.)

934.9ºC

1.694

-4.4

13

15

7

64

1840

10

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CIORWBWIBBPXCG-JAXJKTSHSA-N

InChI=1S/C39H54N10O14S/c1-4-16(2)31-36(60)42-11-29(55)43-25-15-64(63)38-21(20-6-5-18(51)7-22(20)46-38)9-23(33(57)41-12-30(56)47-31)44-37(61)32(17(3)27(53)14-50)48-35(59)26-8-19(52)13-49(26)39(62)24(10-28(40)54)45-34(25)58/h5-7,16-17,19,23-27,31-32,46,50-53H,4,8-15H2,1-3H3,(H2,40,54)(H,41,57)(H,42,60)(H,43,55)(H,44,61)(H,45,58)(H,47,56)(H,48,59)/t16-,17-,19+,23-,24-,25-,26-,27-,31-,32-,64?/m0/s1

2-[(1R,4S,8R,10S,13S,16S,34S)-34-[(2S)-butan-2-yl]-13-[(2R,3R)-3,4-dihydroxybutan-2-yl]-8,22-dihydroxy-2,5,11,14,27,30,33,36,39-nonaoxo-27λ4-thia-3,6,12,15,25,29,32,35,38-nonazapentacyclo[14.12.11.06,10.018,26.019,24]nonatriaconta-18(26),19(24),20,22-tetraen-4-yl]acetamide

α-Amanitin α-Amatoxin

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.  (2). This product is not stable in solution, please use freshly prepared working solution for optimal results.

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

H2O : ~100 mg/mL (~108.82 mM)

Solubility in Formulation 1: 25 mg/mL (27.20 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication (<60°C).

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