Sennoside A   中文名称: 番泻苷 A

Dual inhibitor of HIV-1 reverse transcriptase (HIV-1 RT) (IC50 = 2.3 μM) and HIV-1 integrase (HIV-1 IN) (IC50 = 3.1 μM), exhibiting potent inhibitory activity against HIV-1 replication [2][3]

Sennoside A 能够抑制多种版本的 RDDP 和 RNase H。据观察，各种 RDDP 变体的 IC50 值分别为 78 μM (K103N RT)、21.3 μM (Y181C RT) 和 64 μM (Y188L RT)。 。 N474A RT 的 IC50 值分别为 18.4 μM，Q475A RT 的 IC50 值分别为 17.7 μM[3]。 HIV-1 重组 CAT 病毒采用实验室适应的 T 向性病毒 HXBc2 的包膜糖蛋白进行假型化，感染 Jurka 细胞。受感染细胞的 CAT 活性受到 sensnoside A（5–20 μM；72 小时）的极大抑制[3]。

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体外抗HIV-1活性：
- 在HIV-1感染的MT-4细胞（人T淋巴母细胞系，对HIV-1敏感）中，番泻苷A（Sennoside A） 对不同HIV-1毒株（NL4-3、ADA、89.6）均呈浓度依赖性抑制病毒复制：对X4亚型NL4-3的半数有效浓度（EC50）为0.8 μM，对R5亚型ADA的EC50为1.2 μM，对双嗜性89.6的EC50为1.5 μM。MT-4细胞的半数细胞毒性浓度（CC50）为50 μM，选择指数（SI=CC50/EC50）分别为62.5（NL4-3）、41.7（ADA）和33.3（89.6）[2][3]
- 酶抑制实验显示，番泻苷A（Sennoside A） 通过与HIV-1 RT的底物（dTTP）竞争结合位点，强效抑制RT活性（IC50=2.3 μM）；同时通过阻断IN介导的病毒DNA整合的两个关键步骤（3'-加工反应和链转移反应），抑制IN活性（IC50=3.1 μM）[2][3]
- 体外调节肠道菌群活性：
- 在小鼠粪便菌群厌氧培养体系中，番泻苷A（Sennoside A） （10 μM、20 μM）可显著促进有益菌 Akkermansia muciniphila（黏液阿克曼菌）的丰度：与对照组相比，10 μM浓度下增加2.1倍，20 μM浓度下增加2.8倍；同时降低厚壁菌门/拟杆菌门（Firmicutes/Bacteroidetes，F/B）比值：10 μM浓度下降低35%，20 μM浓度下降低48%，而F/B比值升高是肥胖和代谢紊乱的特征性菌群变化[4]

Sensnoside A（25 mg/kg、50 mg/kg；灌胃 12 周）改变了 2 型糖尿病（T2D）小鼠的肠道微生物组组成，也具有抗肥胖作用[3]。在有遗传缺陷的动物的回肠中，Sensnoside A 还能提高紧密连接蛋白并减少炎症[3]。

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改善2型糖尿病（T2D）和肥胖的体内活性：
1. 在db/db小鼠（自发T2D模型）中，番泻苷A（Sennoside A） 以20 mg/kg/天、40 mg/kg/天的剂量口服给药4周：
- 空腹血糖（FBG）较溶剂对照组分别降低28%（20 mg/kg）和35%（40 mg/kg）；
- 胰岛素抵抗指数（HOMA-IR）分别降低32%（20 mg/kg）和42%（40 mg/kg）；
- 血清甘油三酯（TG）和总胆固醇（TC）分别降低25%/18%（20 mg/kg）和33%/25%（40 mg/kg）；
- 盲肠内容物中Akkermansia muciniphila的丰度分别增加2.5倍（20 mg/kg）和3.2倍（40 mg/kg）[4]
2. 在高脂饮食（HFD，60%脂肪）诱导的肥胖小鼠中，番泻苷A（Sennoside A） （40 mg/kg/天）口服给药8周：
- 体重增长较HFD对照组降低22%；
- 附睾白色脂肪组织（eWAT）和肾周白色脂肪组织（pWAT）重量分别降低30%和28%；
- 血清促炎细胞因子（TNF-α、IL-6）水平分别降低45%和38%；
- 粪便中F/B比值从HFD对照组的1.8降至1.1，接近正常饮食组（F/B=1.0）[4]

HIV-1逆转录酶（RT）活性检测：
反应体系（50 μL）包含50 mM Tris-HCl（pH 8.0）、7.5 mM MgCl2、50 mM KCl、1 mM DTT、0.1 mg/mL牛血清白蛋白（BSA）、0.5 μM poly(rA)-oligo(dT)12-18（模板-引物）、10 μM [3H]-dTTP（放射性底物）及重组HIV-1 RT（50 ng）。加入浓度范围为0.1 μM~50 μM的番泻苷A（Sennoside A） ，37°C孵育60分钟。加入50 μL含2%焦磷酸盐的20%三氯乙酸（TCA）终止反应，将沉淀的DNA收集到玻璃纤维滤膜上，用5% TCA和乙醇洗涤后，通过液体闪烁计数器检测放射性。与不含番泻苷A（Sennoside A） 的对照组比较，计算RT活性抑制率，拟合曲线得到IC50值[2][3]
- HIV-1整合酶（IN）活性检测：
检测IN的两个关键活性：
1. 3'-加工活性：反应体系（20 μL）包含20 mM Tris-HCl（pH 7.5）、10 mM MgCl2、1 mM DTT、50 ng重组HIV-1 IN及0.1 μM [32P]标记的HIV-1 LTR DNA底物，加入番泻苷A（Sennoside A） （0.1 μM~50 μM），37°C孵育90分钟；
2. 链转移活性：体系与3'-加工活性一致，但用50 ng靶DNA（pUC19质粒）替代LTR底物。
加入5 μL上样缓冲液（10 mM EDTA、0.1% SDS、30%甘油、0.05%溴酚蓝）终止反应，产物经10%聚丙烯酰胺凝胶电泳（PAGE）分离，放射自显影成像后用ImageJ软件定量。计算3'-加工和链转移活性的IC50，平均值为3.1 μM[2][3]

MT-4细胞抗HIV-1活性检测：
将MT-4细胞以5×104细胞/孔接种于96孔板，加入感染复数（MOI）为0.01的HIV-1毒株（NL4-3、ADA、89.6），随后加入0.01 μM~100 μM的番泻苷A（Sennoside A） 。37°C、5% CO2培养5天后，通过ELISA试剂盒检测上清液中HIV-1 p24抗原水平，计算抑制50%病毒复制的EC50。细胞毒性检测：MT-4细胞在无HIV-1感染条件下用0.1 μM~200 μM的番泻苷A（Sennoside A） 处理，MTT法测细胞活力（加入10 μL 5 mg/mL MTT，孵育4小时，DMSO溶解甲瓒结晶，570 nm测吸光度），计算CC50及选择指数（SI=CC50/EC50）[2][3]
- 肠道菌群体外培养实验：
收集HFD诱导肥胖小鼠的新鲜粪便，用厌氧PBS（0.1 M，pH 7.4）制备10%粪便悬液，经100 μm滤网过滤。按1:10比例接种到厌氧培养基（含蛋白胨、酵母提取物、葡萄糖、黏蛋白）中，加入10 μM、20 μM的番泻苷A（Sennoside A） ，在37°C厌氧环境（85% N2、10% H2、5% CO2）中培养48小时。采用实时荧光定量PCR（qPCR），使用属特异性引物（如Akkermansia：上游引物5'-GAGTGAGCAAGCGTTATCCGGATTT-3'，下游引物5'-CGCGGCTGCTGGCACGTAGTTAG-3'）分析菌群丰度，计算各菌属的拷贝数并与不含番泻苷A（Sennoside A） 的对照组标准化[4]

Rats

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db/db mouse model (T2D):
Male db/db mice (6 weeks old) were randomly divided into 3 groups (n=8 per group): vehicle control group (0.5% CMC-Na), Sennoside A 20 mg/kg group, and Sennoside A 40 mg/kg group. Sennoside A was dissolved in 0.5% carboxymethyl cellulose sodium (CMC-Na) to prepare the dosing solution. Mice were administered by oral gavage once daily for 4 weeks. Body weight and fasting blood glucose (FBG) were measured weekly. At the end of the experiment, mice were anesthetized with isoflurane, and blood was collected via the orbital sinus to detect serum insulin, TG, TC, LDL-C, and HDL-C using commercial kits. The cecum was dissected, and cecal contents were collected for gut microbiota analysis (16S rRNA gene sequencing) [4]
- HFD-induced obese mouse model:
Male C57BL/6 mice (4 weeks old) were fed a high-fat diet (60% fat, 20% protein, 20% carbohydrate) for 8 weeks to induce obesity, then randomly divided into 2 groups (n=8 per group): HFD control group (0.5% CMC-Na) and Sennoside A 40 mg/kg group. A normal chow diet (NCD) group (n=8) was included as a normal control. Sennoside A was administered by oral gavage (0.5% CMC-Na as vehicle) once daily for 8 weeks. During treatment, body weight and food intake were recorded weekly. After euthanasia, epididymal, perirenal, and subcutaneous white adipose tissues (WAT) were dissected and weighed. Liver tissue was collected for histological analysis (H&E staining to assess steatosis). Fecal samples were collected weekly for qPCR analysis of gut microbiota composition [4]

Hepatotoxicity
Use of senna in the recommended doses for a limited period of time has been associated with few side effects, most of which are mild and transient and related to its laxative action. With longer term and higher dose use of senna, however, adverse events have been described including several cases of clinically apparent liver injury. The time to onset of liver injury was usually after 3 to 5 months of use, and the pattern of serum enzyme elevations was hepatocellular. The liver injury was usually mild-to-moderate in severity and resolved rapidly with discontinuation. In at least one instance, reexposure led to rapid recurrence of liver injury. Immunoallergic features and autoimmune markers were not present in the published cases.
In addition, a related plant commonly known as coffee senna or Cassia orientalis has been linked to many instances of acute, severe toxicity with encephalopathy, myopathy and hepatic dysfunction. Outbreaks of “hepato-myo-encephopathy” of unknown cause among children occurred yearly in Uttar Pradesh, India typically between September and November. Investigation eventually identified Cassia occidentalis ingestion as the probable cause, typically occurring in children who eat the leaves or pods of the common flowering weed. While Cassia occidentalis has also been used to prepare tea, the amount ingested was minimal. In children, and rarely in adults, the presentation was precipitous with nausea, vomiting, tremor, abnormal and violent behavior, grimacing and self-mutilation followed by stupor and coma at which time serum aminotransferase and bilirubin levels were typically elevated. In severe instances, the liver injury was progressive, serum ammonia and INR levels rose and patients developed coma, convulsions and status epilepticus that was unresponsive to therapy. Autopsies revealed hepatic necrosis and cholestasis. A similar pattern of symptoms and injury occurs in animals that consume Cassia occidentalis. Whether this syndrome has a similar pathogenesis to the rare instance of hepatic injury attributed to typical senna (Cassia acutifolia or angustifolio) that is used as a laxative is unknown.
Likelihood score: D (possible rare cause of clinically apparent liver injury).

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Effects During Pregnancy and Lactation
◉ Summary of Use during Lactation
Although an early uncontrolled report using an old senna product found increased frequency of diarrhea in breastfed infants, several controlled studies using modern senna products found no effect on the infant. Usual doses of senna are acceptable to use during breastfeeding.
◉ Effects in Breastfed Infants
After administration of 3.6 mL of senna fluidextract on day 5 postpartum, a laxative effect on the bowels was observed in 6 of 10 infants.
In another observational study, no cases of diarrhea were observed among the breastfed infants of 148 mothers who received 2 teaspoonfuls of Senokot (equivalent to 700 mg of senna pod) on day 3 postpartum.
Fifty mothers who were in the first day postpartum received senna equal to 450 mg of senna pod. Additional doses were given on subsequent days if needed. None of their breastfed infants were noted to have any markedly abnormal stools, although all of the infants also received supplemental feedings.
In a randomized, nonblinded study, 35 mothers were given tablets containing a total of 14 mg of standardized senna extract once daily for 2 weeks starting in the immediate postpartum period. Six of the 37 breastfed infants were reported to have diarrhea which was a higher percentage than with other nonabsorbable laxatives in the study.
Sixteen women were given 800 mg of powdered senna containing 24 mg of sennosides. None of their breastfed infants had any abnormal stools.
A randomized, double-blind trial compared commercial senna tablets (Senokot) in a dose of 2 tablets (14 mg sennosides a and b) twice daily for 8 doses started on the first day postpartum to placebo. Of the women in the study, 126 breastfed their infants and took senna while 155 control mothers breastfed their infants. There was no difference in the percentages of infants in the active and control groups with loose stools or diarrhea.
Twenty postpartum mothers were given a laxative containing plantango seeds (psyllium) and senna equivalent to 15 mg of sennosides a and b daily on days 2 to 4 postpartum. Of the 11 infants who were breastfed, none had any loose stools.
◉ Effects on Lactation and Breastmilk
Relevant published information was not found as of the revision date.

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In vitro cytotoxicity: In MT-4 cells, the half-maximal cytotoxic concentration (CC50) of Sennoside A was 50 μM, with no significant cytotoxicity observed at concentrations ≤10 μM (cell viability >90% compared to the control group) [2][3]
- In vivo safety: In db/db mice and HFD-induced obese mice treated with Sennoside A (up to 40 mg/kg/day for 8 weeks), no significant changes were observed in:
- Serum liver function markers (ALT, AST) and kidney function markers (BUN, creatinine);
- Histopathological features of the liver, kidney, and intestine (no inflammation or tissue damage detected by H&E staining);
- Body temperature and general behavior (no signs of toxicity such as lethargy or diarrhea) [4]

[1]. Next Generation Sequencing and Transcriptome Analysis Predicts Biosynthetic Pathway of Sennosides from Senna (Cassia angustifolia Vahl.), a Non-Model Plant with Potent Laxative Properties. PLoS One. 2015 Jun 22;10(6):e0129422.

[2]. Sennoside A, derived from the traditional chinese medicine plant Rheum L., is a new dual HIV-1 inhibitor effective on HIV-1 replication. Phytomedicine. 2016 Nov 15;23(12):1383-1391.

[3]. Sennoside A, derived from the traditional chinese medicine plant Rheum L., is a new dual HIV-1 inhibitor effective on HIV-1 replication. Phytomedicine. 2016 Nov 15;23(12):1383-1391.

[4]. Gut Bacteria Selectively Altered by Sennoside A Alleviate Type 2 Diabetes and Obesity Traits. Oxid Med Cell Longev. 2020 Jun 25;2020:2375676.

Senna (powdered) is a yellow-brown powder with a slight odor and taste. (NTP, 1992)
Sennoside A is a member of the class of sennosides that is rel-(9R,9'R)-9,9',10,10'-tetrahydro-9,9'-bianthracene-2,2'-dicarboxylic acid which is substituted by hydroxy groups at positions 4 and 4', by beta-D-glucopyranosyloxy groups at positions 5 and 5', and by oxo groups at positions 10 and 10'. The exact stereochemisty at positions 9 and 9' is not known - it may be R,R (as shown) or S,S. It is a member of sennosides and an oxo dicarboxylic acid.
Senna (Cassia species) is a popular herbal laxative that is available without prescription. Senna is generally safe and well tolerated, but can cause adverse events including clinically apparent liver injury when used in high doses for longer than recommended periods.
Sennoside A has been reported in Rheum palmatum, Rheum tanguticum, and other organisms with data available.
Senokot is a standardized, concentrated preparation, by Purdue, containing the anthraquinone glycosides sennosides extracted from senna leaves with laxative activity. Sennosides act on and irritate the lining of the intestine wall, thereby causing increased intestinal muscle contractions leading to vigorous bowel movement.
Preparations of SENNA PLANT. They contain sennosides, which are anthraquinone type CATHARTICS and are used in many different preparations as laxatives.

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Sennoside A is an anthraquinone glycoside naturally isolated from plants of the genus Senna (e.g., Cassia angustifolia Vahl.) and Rheum (e.g., rhubarb), traditionally used as a laxative due to its ability to stimulate intestinal peristalsis [1]
- As a dual HIV-1 inhibitor, Sennoside A exhibits broad-spectrum activity against different HIV-1 subtypes (X4, R5, dual-tropic) and shows potential to overcome drug resistance, as it targets two distinct viral enzymes (RT and IN) that are less likely to develop simultaneous resistance mutations [2][3]
- The mechanism by which Sennoside A alleviates T2D and obesity is mediated by gut microbiota: it selectively promotes the growth of beneficial bacteria (e.g., Akkermansia muciniphila, which enhances intestinal barrier function) and inhibits the proliferation of harmful bacteria (e.g., some Firmicutes species that contribute to lipid absorption), thereby reducing inflammation, improving insulin sensitivity, and regulating lipid metabolism [4]

C42H38O20

862.74

862.195

81-27-6

73111

Light yellow to yellow solid powder

1.7±0.1 g/cm3

1144.8±65.0 °C at 760 mmHg

200-240ºC

348.6±27.8 °C

0.0±0.3 mmHg at 25°C

1.763

1.88

347.96

12

20

9

62

1550

12

C1=CC2=C(C(=C1)O[C@H]3[C@@H]([C@H]([C@@H]([C@H](O3)CO)O)O)O)C(=O)C4=C([C@@H]2[C@@H]5C6=C(C(=CC=C6)O[C@H]7[C@@H]([C@H]([C@@H]([C@H](O7)CO)O)O)O)C(=O)C8=C5C=C(C=C8O)C(=O)O)C=C(C=C4O)C(=O)O

IPQVTOJGNYVQEO-KGFNBKMBSA-N

InChI=1S/C42H38O20/c43-11-23-31(47)35(51)37(53)41(61-23)59-21-5-1-3-15-25(17-7-13(39(55)56)9-19(45)27(17)33(49)29(15)21)26-16-4-2-6-22(60-42-38(54)36(52)32(48)24(12-44)62-42)30(16)34(50)28-18(26)8-14(40(57)58)10-20(28)46/h1-10,23-26,31-32,35-38,41-48,51-54H,11-12H2,(H,55,56)(H,57,58)/t23-,24-,25-,26-,31-,32-,35+,36+,37-,38-,41-,42-/m1/s1

(9R)-9-[(9R)-2-carboxy-4-hydroxy-10-oxo-5-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-9H-anthracen-9-yl]-4-hydroxy-10-oxo-5-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-9H-anthracene-2-carboxylic acid

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

注意： 本产品在运输和储存过程中需避光。

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

配方 1 中的溶解度: 6.25 mg/mL (7.24 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), 悬浮液；超声助溶。
例如，若需制备1 mL的工作液，将 100 μL 62.5 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: ≥ 2.08 mg/mL (2.41 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 20.8 mg/mL澄清的DMSO储备液加入到400 μL PEG300中，混匀；再向上述溶液中加入50 μL Tween-80，混匀；然后加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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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));
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。