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 has the ability to inhibit several versions of RDDP and RNase H. It has been observed that IC50s of 78 μM (K103N RT), 21.3 μM (Y181C RT), and 64 μM (Y188L RT) for various variants of RDDP are inhibited. 18.4 μM for N474A RT and 17.7 μM for Q475A RT, respectively, were the IC50s[3]. HIV-1 recombinant CAT virus, pseudotyped with the envelope glycoprotein of the laboratory-adapted T-tropic virus HXBc2, infects Jurka cells. Infected cells' CAT activity is greatly inhibited by sensnoside A (5–20 μM; 72 h)[3].

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Anti-HIV-1 activity in vitro:
- In HIV-1-infected MT-4 cells (infected with HIV-1 strains NL4-3, ADA, 89.6), Sennoside A showed concentration-dependent inhibition of viral replication. The half-maximal effective concentration (EC50) values were 0.8 μM (NL4-3, X4-tropic), 1.2 μM (ADA, R5-tropic), and 1.5 μM (89.6, dual-tropic). The half-maximal cytotoxic concentration (CC50) in MT-4 cells was 50 μM, resulting in selectivity indices (SI = CC50/EC50) of 62.5 (NL4-3), 41.7 (ADA), and 33.3 (89.6) [2][3]
- In enzyme inhibition assays, Sennoside A potently inhibited HIV-1 RT activity (IC50 = 2.3 μM) by competing with the enzyme’s substrate (dTTP), and inhibited HIV-1 IN activity (IC50 = 3.1 μM) by blocking both the 3'-processing and strand transfer steps of IN-mediated viral DNA integration [2][3]
- Modulation of gut microbiota in vitro:
- In anaerobic cultures of mouse fecal microbiota, Sennoside A (at concentrations of 10 μM and 20 μM) significantly increased the abundance of Akkermansia muciniphila (a beneficial bacterium) by 2.1-fold (10 μM) and 2.8-fold (20 μM) compared to the control group. It also reduced the Firmicutes/Bacteroidetes (F/B) ratio by 35% (10 μM) and 48% (20 μM), which is associated with improved metabolic health [4]

Type 2 diabetes (T2D) mice's gut microbiome composition is changed by sensnoside A (25 mg/kg, 50 mg/kg; intragastric gavage for 12 weeks), which also has anti-obesity effects[3]. In the ileum of genetically faulty animals, sensnoside A also raises tight junction proteins and decreases inflammation[3].

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Efficacy in type 2 diabetes (T2D) and obesity models:
1. In db/db mice (a spontaneous T2D model), oral administration of Sennoside A at doses of 20 mg/kg/day and 40 mg/kg/day for 4 weeks:
- Fasting blood glucose (FBG) decreased by 28% (20 mg/kg) and 35% (40 mg/kg) compared to the vehicle group;
- Homeostasis model assessment of insulin resistance (HOMA-IR) was reduced by 32% (20 mg/kg) and 42% (40 mg/kg);
- Serum triglycerides (TG) and total cholesterol (TC) decreased by 25%/18% (20 mg/kg) and 33%/25% (40 mg/kg), respectively;
- The abundance of Akkermansia muciniphila in the cecum increased by 2.5-fold (20 mg/kg) and 3.2-fold (40 mg/kg) [4]
2. In high-fat diet (HFD)-induced obese mice (60% fat diet for 8 weeks), oral administration of Sennoside A (40 mg/kg/day) for 8 weeks:
- Body weight gain was reduced by 22% compared to the HFD control group;
- Epididymal white adipose tissue (eWAT) and perirenal white adipose tissue (pWAT) weights decreased by 30% and 28%, respectively;
- Serum pro-inflammatory cytokines (TNF-α, IL-6) levels decreased by 45% and 38%, respectively;
- The Firmicutes/Bacteroidetes (F/B) ratio in feces was reduced from 1.8 (HFD control) to 1.1 (Sennoside A group), approaching the normal chow diet group (F/B = 1.0) [4]

HIV-1 reverse transcriptase (RT) activity assay:
The reaction system (50 μL) contained 50 mM Tris-HCl (pH 8.0), 7.5 mM MgCl2, 50 mM KCl, 1 mM DTT, 0.1 mg/mL BSA, 0.5 μM poly(rA)-oligo(dT)12-18 (template-primer), 10 μM [3H]-dTTP (radioactive substrate), and recombinant HIV-1 RT (50 ng). Sennoside A was added at concentrations ranging from 0.1 μM to 50 μM, and the mixture was incubated at 37°C for 60 minutes. The reaction was terminated by adding 50 μL of 20% trichloroacetic acid (TCA) containing 2% pyrophosphate. The precipitated DNA was collected on a glass fiber filter, washed with 5% TCA and ethanol, and the radioactivity was measured using a liquid scintillation counter. The inhibition rate of RT activity was calculated by comparing with the control group (without Sennoside A), and the IC50 value was obtained by curve fitting [2][3]
- HIV-1 integrase (IN) activity assay:
Two key activities of HIV-1 IN were measured:
1. 3'-processing activity: The reaction system (20 μL) contained 20 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 1 mM DTT, 50 ng recombinant HIV-1 IN, and 0.1 μM [32P]-labeled HIV-1 LTR DNA substrate. Sennoside A (0.1 μM–50 μM) was added, and incubation was performed at 37°C for 90 minutes.
2. Strand transfer activity: The system was similar to 3'-processing, but included 50 ng target DNA (pUC19 plasmid) instead of the LTR substrate.
Reactions were terminated by adding 5 μL of loading buffer (10 mM EDTA, 0.1% SDS, 30% glycerol, 0.05% bromophenol blue). Products were separated by 10% polyacrylamide gel electrophoresis (PAGE), visualized by autoradiography, and quantified using ImageJ software. The IC50 values for 3'-processing and strand transfer activities were calculated, with the average IC50 determined as 3.1 μM [2][3]

Anti-HIV-1 activity in MT-4 cells:
MT-4 cells (a human T lymphoblastoid cell line susceptible to HIV-1) were seeded in 96-well plates at 5×104 cells/well. HIV-1 strains (NL4-3, ADA, 89.6) were added at a multiplicity of infection (MOI) of 0.01, followed by Sennoside A at concentrations of 0.01 μM–100 μM. The plates were incubated at 37°C in a 5% CO2 incubator for 5 days. Viral replication was detected by measuring HIV-1 p24 antigen levels in the supernatant using an ELISA kit. The EC50 (concentration inhibiting 50% of viral replication) was calculated. For cytotoxicity assessment, MT-4 cells were treated with Sennoside A (0.1 μM–200 μM) without HIV-1 infection, and cell viability was measured by the MTT assay (adding 10 μL of 5 mg/mL MTT, incubating for 4 hours, dissolving formazan with DMSO, and reading absorbance at 570 nm). The CC50 (concentration causing 50% cell death) and selectivity index (SI = CC50/EC50) were determined [2][3]
- Gut microbiota in vitro culture assay:
Fresh fecal samples were collected from HFD-induced obese mice, suspended in anaerobic PBS (0.1 M, pH 7.4) to prepare a 10% fecal slurry, and filtered through a 100 μm mesh. The slurry was inoculated into anaerobic culture medium (containing peptone, yeast extract, glucose, and mucin) at a 1:10 ratio, and Sennoside A was added at 10 μM and 20 μM. Cultures were incubated at 37°C under anaerobic conditions (85% N2, 10% H2, 5% CO2) for 48 hours. The abundance of gut bacteria was analyzed by quantitative real-time PCR (qPCR) using genus-specific primers (e.g., Akkermansia: forward 5'-GAGTGAGCAAGCGTTATCCGGATTT-3', reverse 5'-CGCGGCTGCTGGCACGTAGTTAG-3'). The copy number of each bacterial genus was calculated and normalized to the control group (without Sennoside A) [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
Short-term use of senna at recommended doses generally results in few side effects, most of which are mild and transient, related to its laxative effect. However, long-term or high-dose use of senna can cause adverse reactions, including several cases of clinically significant liver damage. Liver damage typically appears 3 to 5 months after use, with elevated serum enzymes indicating hepatocellularity. Liver damage is usually mild to moderate and recovers rapidly after discontinuation of the drug. At least one patient experienced a rapid recurrence of liver damage after re-use of senna. No immune hypersensitivity features or autoimmune markers were found in the published cases. Furthermore, a related plant called coffee senna or Oriental Cassia (Cassia orientalis) has also been associated with several cases of acute severe poisoning, including encephalopathy, myopathy, and liver dysfunction. Every year from September to November, Uttar Pradesh, India, experiences outbreaks of unexplained hepatomus-encephalopathy in children. Investigations ultimately determined that ingestion of Cassia occidentalis was likely the cause, typically occurring in children who consumed the leaves or pods of this common flowering weed. Although cassia seeds are also used to make tea, the intake is extremely small. Children (and very rarely adults) present with sudden onset of nausea, vomiting, tremors, unusually violent behavior, facial contortions, and self-harm, followed by coma, at which point serum transaminase and bilirubin levels are usually elevated. In severe cases, liver damage progressively worsens, serum ammonia and international normalized ratio (INR) increase, and patients develop coma, seizures, and status epilepticus, unresponsive to treatment. Autopsy reveals liver necrosis and cholestasis. Animals that ingest senna (Cassia occidentalis) exhibit similar symptoms and damage. Whether the pathogenesis of this syndrome is similar to that of rare liver damage caused by typical senna leaves (Cassia acutifolia or Cassia angustifolio, used as a laxative) is currently unclear. Probability score: D (Possibly a rare cause of clinically significant liver damage).

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Effects during pregnancy and lactation
◉ Overview of medication use during lactation
While an early uncontrolled study using older senna products found an increased incidence of diarrhea in breastfed infants, multiple controlled studies using modern senna products have found no effect on infants. Breastfeeding women can safely use the usual doses of senna.
◉ Effects on breastfed infants
After taking 3.6 ml of senna fluid extract on day 5 postpartum, 6 out of 10 infants developed diarrhea.
In another observational study, 148 mothers took 2 teaspoons of senna extract (equivalent to 700 mg of senna pods) on day 3 postpartum, and none of their breastfed infants experienced diarrhea.
50 mothers took the equivalent of 450 mg of senna pods on day 1 postpartum. Supplemental doses could be given in subsequent days if necessary. No significantly abnormal stools were observed in any of the breastfed infants, although all infants were also supplemented with other feedings. In a randomized, open-label study, 35 mothers began taking tablets containing 14 mg of standardized senna leaf extract once daily for two weeks immediately postpartum. Six out of 37 breastfed infants reported diarrhea, a higher rate than in other non-absorbable laxative groups in the study. Sixteen women took 800 mg of senna leaf powder containing 24 mg of sennosides. None of their breastfed infants experienced abnormal stools. A randomized, double-blind trial compared the effects of commercially available senna leaf tablets (Senokot) (2 tablets twice daily, containing 14 mg of sennosides a and b, for a total of 8 doses, starting from the first postpartum day) with a placebo. Of the participants, 126 mothers breastfed their infants and took senna leaf tablets, while 155 mothers in the control group breastfed their infants. There was no difference in the rate of loose stools or diarrhea between the two groups.
Twenty postpartum mothers took a laxative containing psyllium husk and senna leaves daily from day 2 to 4 postpartum, equivalent to 15 mg of sennosides A and B. None of the 11 breastfed infants experienced loose stools.
◉ Effects on lactation and breast milk
As of the revision date, no relevant published information was found.

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In vitro cytotoxicity: In MT-4 cells, the half-maximal cytotoxic concentration (CC50) of sennoside A was 50 μM, and no significant cytotoxicity was observed at concentrations ≤10 μM (cell viability >90% compared to the control group) [2][3]
In vivo safety: In db/db mice and high-fat diet-induced obese mice, no significant changes were observed in the following after treatment with sennoside A (up to 40 mg/kg/day for 8 weeks):
- Serum liver function indicators (ALT, AST) and kidney function indicators (BUN, creatinine);
- Histopathological features of the liver, kidneys and intestines (no inflammation or tissue damage detected by H&E staining);
- Body temperature and general behavior (no signs of poisoning 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 leaf (powder) is a yellowish-brown powder with a slight odor and taste. (NTP, 1992)
Senna glycoside A belongs to the sennoside class of compounds. Its chemical name is rel-(9R,9'R)-9,9',10,10'-tetrahydro-9,9'-bianthra-2,2'-dicarboxylic acid, in which hydroxyl groups are substituted at the 4 and 4' positions, β-D-glucopyranosyl groups are substituted at the 5 and 5' positions, and oxy groups are substituted at the 10 and 10' positions. The exact stereochemical configuration at the 9 and 9' positions is unknown, and may be R,R (as shown in the figure) or S,S. It belongs to the sennoside class of compounds and is also an oxodicarboxylic acid.
Senna leaf (a plant in the genus Cassia) is a commonly used herbal laxative and can be purchased without a prescription. Senna leaf is generally safe and well-tolerated, but long-term use at high doses may cause adverse reactions, including clinically significant liver damage.
It has been reported that sennoside A is found in rhubarb palmatum, rhubarb tangutica, and other organisms with relevant data.
Senokot is a standardized concentrate produced by Purdue Pharma, containing sennoside, an anthraquinone glycoside extracted from senna leaves, which has laxative activity. Sennoside acts on the intestinal wall and stimulates the intestinal wall lining, thereby causing increased intestinal muscle contractions, ultimately leading to violent defecation.
Senna plant preparations. They contain sennoside, an anthraquinone laxative that is widely used as a laxative in a variety of preparations.

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Sennoside A is an anthraquinone glycoside naturally isolated from plants of the genus Senna (e.g., senna palmatum) and rhubarb (e.g., rhubarb), and has traditionally been used as a laxative because of its ability to stimulate intestinal peristalsis[1].
- As a dual HIV-1 inhibitor, sennoside A has broad-spectrum activity against different HIV-1 subtypes (X4, R5, and bitropic) and shows potential to overcome resistance because it targets two different viral enzymes (reverse transcriptase and integrase), which are unlikely to produce resistance mutations simultaneously [2][3].
- The mechanism by which sennoside A alleviates type 2 diabetes and obesity is mediated by the gut microbiota: it selectively promotes the growth of beneficial bacteria (e.g., Akkermansia mucosae). It enhances intestinal barrier function, inhibits the proliferation of harmful bacteria (e.g., certain Firmicutes that promote 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

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

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

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

Solubility in Formulation 1: 6.25 mg/mL (7.24 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 62.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (2.41 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
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.)