alpha-Amanitin   中文名称: α-鹅膏毒素

在 MKN45 细胞中，α-鹅膏蕈碱可降低 TAF15 mRNA 和蛋白质水平，并阻止 RNAPII 作用于 TAF15 mRNA [2]。在 100、10、1、0.1 和 0.01 μg/mL 剂量下，α-鹅膏蕈碱使细胞活力分别降低 14%、21%、41%、44% 和 50%。 36 小时时，α-鹅膏蕈碱的 LD50 测定为 1 μg/mL。与对照相比，浓度为 1 μg/mL 的 α-鹅膏菌素在 24 小时显着提高了细胞内蛋白质的总量 [3]。当 α-鹅膏蕈碱存在时，卵丘细胞间隙连接基因（Gja1、Gja4 和 Gjc1）以及 FSHr 和 LHr 的表达量要少得多 [4]。

静脉注射后，α-鹅膏蕈碱在BALB/c小鼠体内的LD50为0.327 mg/kg。尾静脉注射α-鹅膏蕈碱后12小时，血清WBC、RBC和HGB水平显着下降，而血清BUN和Crea水平显着升高。某些基因（Hsp90b1、Irx4 等）被 α-鹅膏蕈碱抑制，其编码的蛋白质控制 RNA 聚合酶 II 的活性。某些转录相关蛋白，包括 Nmi 和 Trpc5，会被 α-鹅膏蕈碱下调 [1]。 α-Amanitin 具有很强的 DTC 抑制特性。腹腔注射 MKN45 细胞的小鼠体重持续下降，而注射 α-鹅膏蕈碱 (0.4 mg/kg, ip) 处理的细胞的小鼠体重保持稳定 [2]。

MTT测定用于评估培养细胞的整体功能完整性和活力。将MCF-7细胞放入96孔板中（每孔2×104），孵育24小时。将特定浓度的 α-鹅膏菌素和 β-鹅膏菌素添加到细胞培养基中，并将板再孵育 36 小时。然后将 MTT 溶液（1:10 比例）和二甲基亚砜 (DMSO) (100 µL) 添加到细胞培养基中，并将板孵育过夜。在读板器上在 570 nm 处测量吸光度。该实验重复3次。吸光度数据根据对照组计算为百分比。

Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Amatoxins are water soluble, heat stable polypeptides found in Amanita (most often Amanita phalloides ), Galerina and some Lepiota species. The main toxin from the species A. phalloides is alpha-amanitin, a cyclic octapeptide. It is a potent inhibitor of RNA polymerases that blocks the production of mRNA and protein synthesis in liver and kidney cells. In one case, an infant developed elevated liver enzymes after nursing once 11.5 hours after maternal ingestion of Amanita phalloides. However, two recent, well-documented cases found no adverse effects in breastfed infants and no amatoxin in the milk. Nevertheless, mothers suspected of having Amanita mushroom poisoning probably should not breastfeed until they have recovered or toxicologic screening of the breastmilk has ruled out toxicity.
◉ Effects in Breastfed Infants
In Germany, a 20-year-old nursing mother ate a meal of solely mushrooms identified in the report as green tuberous mushroom (Amanita phalloides). The next morning around 11.5 hours after mushroom ingestion, she nursed her 10-week-old infant who weighed 5 kg. The meal consisted of 80 to 100 mL of breastmilk and the same amount of ready-to-use infant formula (Milasan-Neu). At this time the mother already had symptoms of intoxication (vomiting and diarrhea). Because of the deterioration in her condition she was unable to continue breastfeeding the child, so the child received only formula thereafter. After the mother was admitted to the hospital for Amanita phalloides poisoning and had ASAT and ALAT values of 10,000 and 40,000, respectively (normal values about 500-550). The infant was placed under inpatient observation at children’s hospital. The infant’s relatives noticed nothing unusual about the child and the clinical admission examination revealed no visible evidence of a hepatic, cerebral or hematological disease. Six days after maternal mushroom ingestion, the infant’s laboratory values (electrolytes, serum electrophoresis, bilirubin, gamma-GT, alkaline phosphatase, creatinine, blood sugar, urine status, PTT, and PT [Quick test]) were normal except for ASAT and ALAT, which were about double the normal value. These values slowly decreased and became normal at about day 40 after ingestion.
A 32-year-old mother shared a meal of foraged mushrooms (Amanita bisporigera), and developed symptoms 15 hours post-ingestion. She presented to the emergency department 29 hours post-ingestion and was found to have markedly elevated liver enzymes. Her 4 month-old-daughter had breastfed 4 hours post-ingestion. The asymptomatic infant was evaluated 48 hours after breastfeeding and discharged from the emergency department with no evidence of hepatotoxicity.
A 33-year-old woman picked about 200 mushrooms in a forest in France. She cooked and ate some of them, and developed nausea, vomiting and diarrhea 11 hours post-ingestion. She was admitted to the hospital for treatment and had elevated liver enzymes. She had breastfed her 5-month-old daughter 3 times a day over the 36 hours after mushroom ingestion. Her daughter was hospitalized, but did not present any symptoms, nor any biological disturbance.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

---

Interactions
...PHARMACOLOGICAL AGENTS.../HAVE/ BEEN REPORTED TO PROTECT MICE AGAINST LETHAL DOSES OF PHALLOTOXINS & AMATOXINS /INCLUDING THE AMANITINS/. THE LIST INCLUDES HIGH DOSES OF PENICILLIN, CHLORAMPHENICOL, PHENYLBUTAZONE, & MANY OTHERS... NONE OF THESE ANTIDOTES HAS RECEIVED AN ADEQUATE CLINICAL TRIAL & SO NONE CAN BE RECOMMENDED TODAY. /AMANITINS/
RNA SYNTHESIS IN HELA NUCLEI WAS STIMULATED BY METHYLMERCURY. THIS STIMULATION IS SPECIFIC FOR ALPHA-AMANITIN-SENSITIVE RNA SYNTHESIS (CATALYZED BY RNA POLYMERASE); IN CONTRAST, ALPHA-AMANITIN-RESISTANT SYNTHESIS (CATALYZED BY RNA POLYMERASES I & III) WAS 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-Amanitin has been reported in Amanita suballiacea, Amanita phalloides, and other organisms with data available.
A cyclic octapeptide with a thioether bridge between the cystine and tryptophan. It inhibits RNA POLYMERASE II. Poisoning may require LIVER TRANSPLANTATION.

---

Mechanism of Action
/IN COMPARISON WITH THE PHALLOTOXINS/...THE LONG DELAYED HEPATOTOXIC RESPONSE SEEN IN HUMAN POISONINGS...IS MORE LIKELY DUE TO...ALPHA-, BETA-, & GAMMA-AMANITIN, ESPECIALLY THE ALPHA COMPONENT. THESE SO-CALLED AMATOXINS...ARE MORE TOXIC THAN THE PHALLOTOXINS, &, UNLIKE THE LATTER, THEY DAMAGE THE NUCLEOLUS & LATER THE NUCLEUS OF LIVER CELLS.

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

400.09

13

15

7

64

1840

10

CC[C@H](C)[C@H]1C(=O)NCC(=O)N[C@H]2CS(=O)C3=C(C[C@@H](C(=O)NCC(=O)N1)NC(=O)[C@@H](NC(=O)[C@@H]4C[C@H](CN4C(=O)[C@@H](NC2=O)CC(=O)N)O)[C@@H](C)[C@H](CO)O)C5=C(N3)C=C(C=C5)O

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

注意： (1). 本产品在运输和储存过程中需避光。  (2). 该产品在溶液状态不稳定，请现配现用。

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)

配方 1 中的溶解度: 25 mg/mL (27.20 mM) in PBS (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。 (<60°C).

---

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));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
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；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。