SIRT1 0.10 μM (EC50)
SRT 1720 selectively targets SIRT1 (sirtuin 1), a class III histone deacetylase that requires NAD⁺ as a cofactor for its enzymatic activity. SIRT1 deacetylates various substrates including histones, transcription factors (such as PGC-1α, FOXO, and NF-κB), and other proteins involved in cellular metabolism, stress resistance, and longevity. The compound acts as an allosteric activator of SIRT1, enhancing its deacetylase activity. SRT 1720 shows much weaker activity against SIRT2 and SIRT3, with EC50 values of 37 μM and 300 μM respectively, indicating high selectivity for SIRT1. The activation of SIRT1 by SRT 1720 leads to downstream effects on mitochondrial biogenesis, fatty acid oxidation, and inflammatory responses.

SRT 1720 dihydrochloride, which is linked to the histone acetyltransferase p300, efficiently lowers p53 acetylation in cells, even in the absence of SIRT1 [3].

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SRT1720 is a selective activator of SIRT1 with an EC50 of 0.16 μM, which is more than 230 times lower than SIRT2 and SIRT3.
Here, researchers examined the anti-multiple myeloma (MM) activity of a novel oral agent, SRT1720, which targets SIRT1. Treatment of MM cells with SRT1720 inhibited growth and induced apoptosis in MM cells resistant to conventional and bortezomib therapies without significantly affecting the viability of normal cells. Mechanistic studies showed that anti-MM activity of SRT1720 is associated with: 1) activation of caspase-8, caspase-9, caspase-3, poly (ADP) ribose polymerase; 2) increase in reactive oxygen species; 2) induction of phosphorylated ataxia telangiectasia mutated/checkpoint kinase 2 signalling; 3) decrease in vascular endothelial growth factor-induced migration of MM cells and associated angiogenesis; and 4) inhibition of nuclear factor-κB. Blockade of ATM attenuated SRT1720-induced MM cell death. [2]

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To determine whether SRT1460 and SRT1720 bind and activate the enzyme at the same molecular site as resveratrol, an isobologram analysis was performed. A concentration matrix of two compounds, resveratrol versus SRT1720 and SRT1720 versus SRT1460, was examined to determine whether the combination was antagonistic, additive or synergistic. In both cases the compound combination resulted in additivity consistent with the hypothesis that a single allosteric site exists on the SIRT1—substrate complex to which structurally diverse compounds can bind (Fig. 2c). [1]
In vitro, SRT 1720 has been shown to activate SIRT1-mediated deacetylation of various substrates. At 1 μM concentration, the compound reverses UVA-inhibited expression of PGC-1α, NRF2, and TFAM in B4G12 cells. SRT 1720 effectively decreases the levels of cell death and ROS production in cellular models of oxidative stress. The compound promotes mitochondrial biogenesis and enhances fatty acid oxidation in cultured myotubes and adipocytes. In hepatocytes, SRT 1720 reduces lipid accumulation and improves insulin sensitivity through SIRT1-dependent mechanisms. The compound also exhibits anti-inflammatory effects by inhibiting NF-κB signaling and reducing pro-inflammatory cytokine production in macrophages. SRT 1720 shows cytoprotective effects in various cell types under stress conditions.

Lepob/ob mice with SRT 1720 (10, 30, 100 mg/kg, po) dihydrochloride therapy had a considerable reduction in fasting blood glucose to values close to normal [1]. Through downstream targets of SIRT1, such as PGC1α and FOXO1, SRT 1720 dihydrochloride protects mice against the harmful effects of diet-induced obesity and is linked to metabolic adaptation of fatty acid and oxidative metabolism [3]. In wild-type rats, SRT 1720 (50–100 mg/kg, po) dihydrochloride decreases arterial oxygen saturation and attenuates elastase-induced airway expansion and lung function degradation during emphysema development [4].

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In DIO mice SRT1720 mimics several of the effects observed after calorie restriction including improved insulin sensitivity, normalized glucose and insulin levels, and increased mitochondrial capacity. In addition, in diet-induced obese and genetically obese mice, SRT1720 improves insulin sensitivity, lower plasma glucose, and increase mitochondrial capacity. Thus, SRT1720 is a promising new therapeutic agent for treating diseases of ageing such as type 2 diabetes. Consistent with improved glucose tolerance, the glucose infusion rate required to maintain euglycaemia is approximately 35% higher in SRT1720-treated fa/fa rats, and the total glucose disposal rate is increased by approximately 20%. SRT1720 also prevents multiple myeloma tumor growth. SRT1720 increases the cytotoxic activity of bortezomib or dexamethasone.

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SIRT1 activation by both genetic overexpression and a selective pharmacological activator, SRT1720, attenuated stress-induced premature cellular senescence and protected against emphysema induced by cigarette smoke and elastase in mice. Ablation of Sirt1 in airway epithelium, but not in myeloid cells, aggravated airspace enlargement, impaired lung function, and reduced exercise tolerance. These effects were due to the ability of SIRT1 to deacetylate the FOXO3 transcription factor, since Foxo3 deficiency diminished the protective effect of SRT1720 on cellular senescence and emphysematous changes. [4]

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However, whether SIRT1 is a suitable therapeutic target for the treatment of cholestasis is unknown. In the present study, researchers examined the protective effect of SRT1720, which is a specific activator of SIRT1, against 17α-ethinylestradiol (EE)-induced cholestasis in mice. The data demonstrated that SRT1720 significantly prevented EE-induced changes in the serum levels of total bile acids (TBA), total bilirubin (TBIL), γ-glutamyltranspeptidase (γ-GGT) and alkaline phosphatase (ALP). SRT1720 also relieved EE-induced liver pathological injuries as indicated by haematoxylin and eosin (H&E) staining. SRT1720 treatment protected against EE-induced liver injury through the HNF1α/FXR signalling pathway, which up-regulated the expression of hepatic efflux transporter (Bsep and Mrp2) and hepatic uptake transporters (Ntcp and Oatp1b2). Moreover, SRT1720 significantly inhibited the TNF-α and IL-6 levels induced by EE. These findings indicate that SRT1720 exerts a dose-dependent protective effect on EE-induced cholestatic liver injury in mice and that the mechanism underlying this activity is related to the activation of the HNF1α/FXR signalling pathway and anti-inflammatory mechanisms.[5]
In vivo, SRT 1720 has demonstrated beneficial metabolic effects in various animal models. In diet-induced obese mice, oral administration of SRT 1720 improves insulin sensitivity, reduces hepatic steatosis, and enhances glucose tolerance. The compound increases mitochondrial function and fatty acid oxidation in skeletal muscle and liver tissues. In mouse models of aging, SRT 1720 extends healthspan and improves metabolic parameters. The compound has also shown protective effects in cardiovascular disease models, reducing cardiac hypertrophy and improving heart function. In models of neuroinflammation and neurodegeneration, SRT 1720 provides neuroprotective effects through SIRT1 activation. The compound is typically administered orally in preclinical studies at doses ranging from 10 to 100 mg/kg.

SIRT1 Fluorescence Polarization Assay and HTS [1]
In the SIRT1 FP assay, SIRT1 activity was monitored using a 20 amino acid peptide (AcGlu-Glu-Lys(biotin)-Gly-Gln-Ser-Thr-Ser-Ser-His-Ser-Lys(Ac)-Nle-Ser-Thr-Glu-Gly– Lys(MR121 or Tamra)-Glu-Glu-NH2) derived from the sequence of p53. The peptide was N-terminally linked to biotin and C-terminally modified with a fluorescent tag. The reaction for monitoring enzyme activity was a coupled enzyme assay where the first reaction was the deacetylation reaction catalyzed by SIRT1 and the second reaction was cleavage by trypsin at the newly exposed lysine residue. The reaction was stopped and streptavidin was added in order to accentuate the mass differences between substrate and product. In total, 290,000 compounds were screened and 127 hits were confirmed. The sensitivity of the FP assay allowed identification of compounds that exhibited low level activation of SIRT1 (≥17% activation at 20 μM) producing multiple classes of activators representing distinct structural classes. The fluorescence polarization reaction conditions were as follows: 0.5 μM peptide substrate, 150 μM βNAD+ , 0-10 nM SIRT1, 25 mM Tris-acetate pH 8, 137 mM Na-Ac, 2.7 mM K-Ac, 1 mM Mg-Ac, 0.05% Tween-20, 0.1% Pluronic F127, 10 mM CaCl2, 5 mM DTT, 0.025% BSA, and 0.15 mM nicotinamide. The reaction was incubated at 37°C and stopped by addition of nicotinamide, and trypsin was added to cleave the deacetylated substrate. This reaction was incubated at 37o C in the presence of 1 μM streptavidin. Fluorescent polarization was determined at excitation (650 nm) and emission (680 nm) wavelengths.

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Mechanism of action studies [1]
The effect of test compounds on the Km of human SIRT1 enzyme for acetylated peptide substrate was examined using the SIRT1 mass spectrometry assay described above. Using the cell-free MS assay, the Km of SIRT1 enzyme for peptide substrate was determined at nine concentrations of compound (100, 33, 11, 3.7, 1.2, 0.41, 0.14, 0.046, and 0.015 μM) and also in the presence of DMSO vehicle alone. To determine the Km, the linear deacetylation rate was determined at 12 concentrations of acetylated peptide substrate (50, 25, 12.5, 6.25, 3.12, 1.56, 0.78, 0.39, 0.19, 0.098, 0.049, and 0.024 μM) for each of the compound concentrations and for the vehicle control. SIRT1 enzyme, 2 mM NAD+ , and 0-50 μM acetylated peptide substrate were incubated with 0-100 μM compound at 25°C. At 0, 3, 6, 9, 12, 15, 20, and 25 minutes, the reaction was stopped with 10% formic acid with 50 mM nicotinamide and the conversion of substrates to products determined by mass spectrometry.

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Isothermal titration Calorimetry (ITC) [1]
The human SIRT1-E5c protein (41 μM; described above), the mass spectrometry peptide substrate (1.0 mM), and SRT1460 (0.84 mM) stock solutions were used for the ITC. The buffer conditions were 50 mM Tris-HCl (pH 8.0), 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2, 2 mM TCEP, and 5% glycerol. Titrations were carried out at 26ºC on a VP-ITC. SRT1460 was selected for these studies because it is soluble in buffer to the millimolar concentrations required in the experiment.

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Isobologram studies [1]
The effect of the combination of resveratrol versus SRT1720 and SRT1720 versus SRT1460 was determined using the SIRT1 mass spectrometry assay described above. A concentration matrix of the 2 compounds was created and tested against SIRT1 enzyme. The % conversion of acetylated peptide substrate to deacetylated peptide product was determined at each of the combinations present in the matrix. The resulting Isobologram was used to evaluate the effect of the combination. For the analysis, a plot in Cartesian coordinates of a dose combination that produce the same effect level is the basis for an Isobologram. If two compounds have variable potency, a constant relative potency (R) - which is the amount of compound needed to achieve the same fold activity (e.g. EC1.25 for resveratrol vs SRT1720 and EC2.5 for SRT1720 vs SRT1460) - is selected for the X and Y intercepts for Isobologram analysis. The concentration of both compounds which corresponds to the respective EC value is used as an intercept on both the X and Y axes. Using these two intercepts, a theoretical line called the line of additivity is drawn between the two points. Experimental data obtained by the logarithmic titration of the two compounds mixed as a dose pair in a matrix which yield the same effect level (EC value), is plotted on the Isobologram. Statistical comparison of the line of additivity and the curve arising from experimental two drug dose combinations indicates if an effect is additive. Points falling below and above the line of additivity are subjected to regression analysis. Experimental data that is higher than the line of additivity is interpreted antagonistic and experimental data that is lower is interpreted as synergistic, and experimental data that fall on the line of additivity is considered additive.

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p53 Deacetylation Assay [1]
Human osteosarcoma cells (U-2 OS) were plated at 1.5 X 104 cells per well in 96 well plates. Test compounds and controls (all in 100% DMSO) were added to cell plates after 24 h. To demonstrate the SIRT1-dependence of this assay read-out, replicate plate sets were co-treated with both test compounds and a SIRT1-specific small molecule inhibitor, 6-chloro-2,3,4,9-tetrahydro-1-H-carbazole-1-carboxamide. After compound addition, p53 expression and acetylation was induced by the addition of doxorubicin (1 μg/ml final concentration) to each well. Following p53 induction, cells were fixed and then permeablized with PBS-0.1% Triton-X-100. Non-specific protein binding was then blocked by addition of a solution of 5% BSA in PBS-0.1% TWEEN 20 (Block Solution). Primary antibodies, anti-p53-acetyl-lysine-382 (rabbit polyclonal) and anti-beta tubulin (mouse monoclonal), were diluted 1:400 and 1:1000, respectively, in Block Solution and added to wells for an overnight incubation at 4o C. Wells were then washed in PBS-0.1% TWEEN 20 (Wash Buffer). Secondary antibodies, IR800CW Goat anti-rabbit IgG and Alexa-Fluor 680 goat anti-mouse IgG , were diluted in Block Solution and added to wells for a 1 hr, room temperature incubation. Wells were again washed 5 times in Wash Buffer. Plates were then scanned with a Li-Cor Odyssey infrared scanner. Data was extracted using manufacturer’s software. Signals for both Ac-Lys382-p53 and beta-tubulin were background corrected using wells incubated with only secondary antibodies. Each well Ac-Lys382-signal was then normalized to its corresponding beta-tubulin signal to correct for differences in cell number. These values were then normalized to vehicle to generate the % acetylated p53 value for each well.
In vitro enzyme assays for SIRT1 activation by SRT 1720 are performed using purified recombinant SIRT1 enzyme and a fluorogenic peptide substrate. The assay is conducted in a buffer containing NAD⁺, the peptide substrate, and varying concentrations of the compound. The reaction is incubated at 37 °C for a specified period, and the deacetylated product is detected using a fluorescence-based or luminescence-based readout. The EC50 value is determined from the concentration-response curve. For selectivity assessment, similar assays are performed using SIRT2 and SIRT3 enzymes. The assay includes positive controls such as resveratrol and negative controls (vehicle only). All reactions are performed in triplicate with appropriate blanks and standards.

In situ detection of apoptosis using immunohisto-chemistry (IHC) [2]
Tumours from vehicle (control)- and SRT1720-treated mice were excised and preserved in 10% formalin. Apoptotic cells in tumours were identified by IHC staining for caspase-3 activation, as previously described (Chauhan et al, 2010).

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In vitro migration and capillary-like tube structure formation assays [2]
Transwell Insert Assays were utilized to measure migration as previously described (Podar et al, 2001). In vitro angiogenesis was assessed by Matrigel capillary-like tube structure formation assay (Chauhan et al, 2010). For endothelial tube formation assay, human vascular endothelial cells (HUVECs) were obtained from Clonetics and maintained in endothelial cell growth medium-2 (EGM2 MV SingleQuots) containing 5% FBS. After three passages, HUVEC cell viability was measured with the trypan blue exclusion assay, and <5% of cell death was observed with SRT1720 treatment.

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Cell viability and apoptosis assays[2]
Cell viability was assessed with a colorimetric assay using 3-(4, 5-dimethylthiozol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) as described previously (Hideshima et al, 2000). Apoptosis assay was quantified using Annexin V-FITC/Propidium iodide (PI) apoptosis detection kit, as per manufacturer’s instructions , followed by analysis on FACS Calibur.

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Western blotting and protein quantification[2]
Immunoblot analysis was performed using antibodies against caspase-3, caspase-7, caspase-8, caspase-9, poly (ADP) ribose polymerase (PARP), Ace-Lys 382 p53, phosphorylated-ataxia telangiectasia mutated (pATM), phosphorylated- checkpoint kinase 2 (pCHK2), phosphorylated-IκB ser32/36, and GAPDH. Blots were then developed by enhanced chemiluminescence. Densitometry of protein bands was acquired using an AlphaImager EC gel documentation system, and bands were analysed using the spot densitometry analysis tool.
In vitro cell-based assays for SRT 1720 typically use cell lines such as HepG2 hepatocytes, C2C12 myotubes, or 3T3-L1 adipocytes to assess SIRT1 activation and downstream metabolic effects. Cells are treated with SRT 1720 at concentrations ranging from 0.1 to 10 μM for 24-72 hours. SIRT1 activity is measured by assessing the acetylation status of known SIRT1 substrates (e.g., PGC-1α, FOXO1, p53) using Western blotting with specific antibodies. Mitochondrial function is evaluated by measuring oxygen consumption rate (OCR) and ATP production. Gene expression of SIRT1 target genes is assessed by qRT-PCR. Cell viability is monitored using MTT or CCK-8 assays. Apoptosis is evaluated by flow cytometry or caspase activity assays.

Dissolved in 20% PEG400/0.5% Tween80/79.5% deionized water; 200 mg/kg/day; Oral administration

The pharmacokinetic characteristics of SRT1720 (Figure 3a) are applicable to in vivo evaluation in mice (bioavailability = 50%, terminal half-life approximately 5 hours, area under the curve (AUC) = 7,892 ng h−1 ml−1) and rats (bioavailability = 25%, terminal half-life approximately 8.4 hours, AUC = 3,714 ng h−1 ml−1). SRT501 is a modified version of resveratrol with higher bioavailability (bioavailability of 11%, terminal half-life approximately 1 hour, AUC of 10,524 ng h−1 ml−1) and has also been studied in genetically obese mice (Lepob/ob) and diet-induced obese (DIO) mice. [1]
SRT 1720 is orally bioavailable with good pharmacokinetic properties. Following oral administration, the compound is rapidly absorbed and distributed to various tissues including liver, muscle, and adipose tissue. The compound exhibits dose-proportional pharmacokinetics with a terminal half-life suitable for once-daily dosing. SRT 1720 is metabolized primarily in the liver through cytochrome P450 enzymes, with major metabolites being excreted in bile and urine. The compound shows moderate protein binding in plasma. Pharmacokinetic studies in rodents have demonstrated that SRT 1720 achieves sufficient tissue concentrations to activate SIRT1 in target organs. The compound's oral bioavailability and favorable pharmacokinetic profile support its use in in vivo efficacy studies.

SRT 1720 is generally well-tolerated in preclinical studies with no significant toxicity observed at therapeutic doses. In rodent studies, the compound has shown a favorable safety profile with no major organ toxicity or hematological abnormalities. The no-observed-adverse-effect level (NOAEL) has been established at doses several-fold higher than the efficacious doses. No genotoxicity or carcinogenicity has been reported in standard assays. However, as with all research chemicals, appropriate safety precautions should be taken when handling SRT 1720. The compound should be stored at -20°C and protected from light. Safety data sheets recommend using personal protective equipment including gloves and safety goggles. The compound is for research use only and not for human or veterinary use.

[1]. Small molecule activators of SIRT1 as therapeutics for the treatment of type 2 diabetes. Nature. 2007 Nov 29;450(7170):712-6.
[2]. Preclinical evaluation of a novel SIRT1 modulator SRT1720 in multiple myeloma cells. Br J Haematol. 2011 Dec;155(5):588-98.

[3]. Are sirtuins viable targets for improving healthspan and lifespan?,Nat Rev Drug Discov. 2012 Jun 1;11(6):443-61.

[4]. SIRT1 protects against emphysema via FOXO3-mediated reduction of premature senescence in mice.,J Clin Invest. 2012 Jun 1;122(6):2032-45.

[5]. Activation of SIRT1 Attenuates Klotho Deficiency-Induced Arterial Stiffness and Hypertension by Enhancing AMP-Activated Protein Kinase Activity. Hypertension. 2016 Nov;68(5):1191-1199.

[6]. SRT1720 induces lysosomal-dependent cell death of breast cancer cells. Mol Cancer Ther. 2015 Jan;14(1):183-92.

[7]. Protective effects of SRT1720 via the HNF1α/FXR signalling pathway and anti-inflammatory mechanisms in mice with estrogen-induced cholestatic liver injury. Toxicol Lett. 2016 Dec 15;264:1-11.

Caloric restriction can extend lifespan and produce metabolic characteristics beneficial for the treatment of age-related diseases such as type 2 diabetes. SIRT1, an NAD+-dependent deacetylase, is a major regulator of downstream pathways of caloric restriction, which play a beneficial role in glucose homeostasis and insulin sensitivity. Resveratrol, a polyphenolic SIRT1 activator, mimics the anti-aging effects of caloric restriction in lower organisms and mice fed a high-fat diet, improving insulin resistance, increasing mitochondrial content, and extending lifespan. This article describes the identification and characterization of small molecule SIRT1 activators that are structurally independent of resveratrol but exhibit 1000-fold higher activity. These compounds bind to the allosteric site at the N-terminus of the catalytic domain of the SIRT1 enzyme-peptide substrate complex and reduce the Michaelis constant of the acetylated substrate. In diet-induced obese mice and genetically obese mice, these compounds improved insulin sensitivity, reduced plasma glucose levels, and enhanced mitochondrial function. In Zucker fa/fa rats, the hyperinsulin-eukemic clamp test showed that SIRT1 activators improved systemic glucose homeostasis and increased insulin sensitivity in adipose tissue, skeletal muscle and liver. Therefore, SIRT1 activation is a promising new approach for treating age-related diseases such as type 2 diabetes. [1] SIRT1 belongs to the Silent Information Regulator 2 (Sir2) protein family and is an NAD(+)-dependent class III histone deacetylase. This study investigated the anti-multiple myeloma (MM) activity of a novel oral drug, SRT1720, which targets SIRT1. Treatment of multiple myeloma (MM) cells with SRT1720 inhibited the growth of MM cells resistant to conventional therapy and bortezomib and induced their apoptosis, without significantly affecting the viability of normal cells. Mechanistic studies revealed that the anti-MM activity of SRT1720 is associated with the following mechanisms: (i) activation of caspase-8, caspase-9, caspase-3, and poly(ADP)ribose polymerase; (ii) increased reactive oxygen species; (iii) induction of phosphorylated ataxia-telangiectatic mutant protein/checkpoint kinase 2 (ATM/CK2) signaling pathway; (iv) reduction of vascular endothelial growth factor (VEGF)-induced MM cell migration and related angiogenesis; and (v) inhibition of nuclear factor-κB (NF-κB). Blocking ATM attenuated SRT1720-induced MM cell death. In animal tumor models, SRT1720 inhibited MM tumor growth. Finally, SRT1720 enhanced the cytotoxic activity of bortezomib or dexamethasone. Our preclinical studies provide a theoretical basis for novel therapies targeting SIRT1 in multiple myeloma (MM). [2]

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While the increase in human lifespan reflects the success of modern medicine, the risk of developing a variety of diseases also increases exponentially with age. Caloric restriction is known to delay aging, slow functional decline, and delay the onset of most diseases in organisms. Studies have shown that sirtuins (SIRT1-SIRT7) are mediators of the key effects of caloric restriction during aging. Two unrelated molecules—resveratrol and SRT1720—have been shown to enhance SIRT1 activity under certain conditions. They are excellent protectants against metabolic stress in mammals, making SIRT1 a potential and attractive therapeutic target. This review covers the current status and controversies of sirtuins as novel pharmacological targets, with a focus on SIRT1. [3]

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Chronic obstructive pulmonary disease/emphysema (COPD/emphysema) is characterized by chronic inflammation and premature aging of the lungs. The anti-aging protein sirtuin 1 (SIRT1) is an NAD+-dependent protein/histone deacetylase that is reduced in expression in the lungs of COPD patients. However, the molecular mechanisms underlying premature lung aging, and whether SIRT1 can counteract the various pathophysiological changes associated with cellular senescence and emphysema, remain unknown. In this study, we found increased cellular senescence in the lungs of COPD patients. Activation of SIRT1, whether through gene overexpression or the selective pharmacological activator SRT1720, alleviated stress-induced premature cellular senescence and protected mice from cigarette smoke and elastase-induced emphysema. Knockout of SIRT1 in airway epithelial cells (but not myeloid cells) exacerbated alveolar enlargement, impaired lung function, and reduced exercise tolerance. These effects are attributed to SIRT1's ability to deacetylate the FOXO3 transcription factor, as Foxo3 deficiency attenuates the protective effect of SRT1720 against cellular senescence and emphysema changes. Suppressing lung inflammation with NF-κB/IKK2 inhibitors did not benefit emphysema. Therefore, SIRT1 protects against emphysema through FOXO3-mediated reduction of cellular senescence, independent of inflammation. Activation of SIRT1 may be a promising strategy for treating chronic obstructive pulmonary disease/emphysema. [4]

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Sirtuin 1 (SIRT1) is the most conserved NAD+-dependent protein deacetylase in mammals and belongs to the silencing information regulator 2 (Sir2) protein family (also known as Sirtuins). In the liver, hepatic SIRT1 regulates bile acid metabolism by modulating the expression of farnesol X receptor (FXR). FXR is one of the most important nuclear receptors involved in the regulation of bile acid metabolism. SIRT1 regulates FXR expression at multiple levels, including direct deacetylation of the transcription factor and transcriptional regulation via hepatocyte nuclear factor 1α (HNF1α). Therefore, hepatic SIRT1 is an important regulator of the HNF1α/FXR signaling pathway and hepatic bile acid metabolism. However, it is unclear whether SIRT1 is a suitable target for the treatment of cholestasis. This study investigated the protective effect of the SIRT1-specific activator SRT1720 against 17α-ethinylestradiol (EE)-induced cholestasis in mice. The results showed that SRT1720 significantly inhibited changes in serum total bile acid (TBA), total bilirubin (TBIL), gamma-glutamyl transferase (γ-GGT), and alkaline phosphatase (ALP) levels induced by EE. Hematoxylin-eosin (H&E) staining results showed that SRT1720 also alleviated EE-induced liver pathological damage. SRT1720 exerted a protective effect through the HNF1α/FXR signaling pathway, upregulating the expression of hepatic efflux transporters (Bsep and Mrp2) and hepatic uptake transporters (Ntcp and Oatp1b2), thereby alleviating EE-induced liver injury. In addition, SRT1720 significantly inhibited EE-induced TNF-α and IL-6 levels. These results indicate that SRT1720 has a dose-dependent protective effect against EE-induced cholestatic liver injury in mice, and that the mechanism of this effect is related to the activation of the HNF1α/FXR signaling pathway and anti-inflammatory mechanisms. [5]
SRT 1720 is one of the most widely studied SIRT1 activators in the research community and has been cited in numerous publications in high-impact journals. The compound was developed as part of a series of synthetic sirtuin-activating compounds (STACs) that mimic the effects of calorie restriction and resveratrol. SRT 1720 has been instrumental in elucidating the role of SIRT1 in metabolic regulation, aging, and disease pathogenesis. While the compound has shown promising preclinical results, it has not been approved for clinical use. Other SIRT1 activators in the same class have been evaluated in clinical trials for metabolic and aging-related diseases. SRT 1720 remains a valuable research tool for studying SIRT1 biology and developing novel therapeutic strategies targeting sirtuin pathways.

C25H24CLN7OS

542.48

541.121

2468639-77-0

SRT 1720;925434-55-5;SRT 1720 monohydrochloride;2060259-60-9; 2468639-77-0 (2HCl); 1001645-58-4 (x HCl)

139026900

Brown to orange solid powder

4

7

5

36

707

0

C1CN(CCN1)CC2=CSC3=NC(=CN23)C4=CC=CC=C4NC(=O)C5=NC6=CC=CC=C6N=C5.Cl.Cl

YBWQTKUVUFMWOX-UHFFFAOYSA-N

InChI=1S/C25H23N7OS.2ClH/c33-24(22-13-27-20-7-3-4-8-21(20)28-22)29-19-6-2-1-5-18(19)23-15-32-17(16-34-25(32)30-23)14-31-11-9-26-10-12-31;;/h1-8,13,15-16,26H,9-12,14H2,(H,29,33);2*1H

N-[2-[3-(piperazin-1-ylmethyl)imidazo[2,1-b][1,3]thiazol-6-yl]phenyl]quinoxaline-2-carboxamide;dihydrochloride

SRT 1720 dihydrochloride; SRT 1720 (dihydrochloride); 2468639-77-0; HY-15145A; AKOS037515640; DA-78016; MS-29982; N-[2-[3-(piperazin-1-ylmethyl)imidazo[2,1-b][1,3]thiazol-6-yl]phenyl]quinoxaline-2-carboxamide;dihydrochloride;

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 : 85 mg/mL (156.69 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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