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| 5mg |
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Purity: ≥98%
SBI-115 (SBI115) is a novel and potent TGR5 (GPCR19) antagonist with the potential to be used for polycystic liver disease. In polycystic liver diseases, it reduces hepatic cystogenesis by blocking TGR5. By boosting cholangiocyte proliferation and cAMP levels, TGR5 aids in the process of hepatic cystogenesis1. A TGR5 antagonist, either by itself or in combination with somatostatin receptor agonists, may be a useful treatment option for polycystic liver disease. TGR5 promotes hepatic cystogenesis by elevating cAMP and cholangiocyte proliferation.
| Targets |
TGR5
TGR5 (G protein-coupled bile acid receptor). [1] |
|---|---|
| ln Vitro |
SBI-115 (100-200 μM, 24 hours) prevents the proliferation of shRNA-transfected ADPKD cholangiocytes that is caused by pre-treating cystic cholangiocytes with taurolithocholic acid (TLCA)[1].
In cystic cholangiocytes derived from ADPKD patients, SBI-115 (100 and 200 µM) dose-dependently inhibited TLCA-induced cell proliferation by 32-48%, as assessed by MTS assay and cell counting. [1] SBI-115 (200 µM) reduced the growth of cholangiocyte spheroids formed by ADPKD cholangiocytes in 3-D hanging drop culture by approximately 30%. [1] SBI-115 (200 µM) decreased cAMP levels in ADPKD cystic cholangiocytes that had been pre-stimulated with the TGR5 agonist TLCA by approximately 30%. This effect was absent in TGR5-depleted (via shRNA) cholangiocytes, confirming TGR5-dependent action. [1] Concurrent treatment of ADPKD cystic cholangiocytes with SBI-115 (200 µM) and pasireotide (20 µM, an SSTR agonist) led to a greater reduction in cell proliferation (45-55% inhibition), spheroid growth, and cAMP levels compared to each drug alone. [1] SBI-115 alone had no effect on cell proliferation, spheroid growth, or cAMP levels in the absence of TGR5 agonist stimulation. [1] SBI-115 had no effect on forskolin-induced cAMP production in cystic cholangiocytes, indicating its action is specific to TGR5-mediated cAMP signaling. [1] |
| ln Vivo |
In order to further investigate TGR5's role in PLD, we produced double-mutant mice with the TGR5−/−;Pkhd1del2/del2 phenotype. TGR5−/− mice demonstrated good health and reproducibility, in line with an earlier report. 29, Overall morphology (Supporting Fig. 4), liver, cholangiocyte, and primary cilium morphology (Supporting Table 4), and serum biochemistries (Supporting Table 3) were similar in WT and TGR5−/− mice. More than one hepatic cyst is present in Pkhd1del2/del2 rodents. 30, TGR5 was overexpressed in Pkhd1del2/del2 mice relative to WT, while it was not seen in TGR5−/− mice or TGR5−/−;Pkhd1del2/del2 mice, which is in line with our findings in other animal models of PLD (Supporting Fig. 5). Compared to Pkhd1del2/del2 littermates, we observed reductions in liver weight (30%), hepatic cystic areas (31%), and hepatic fibrotic areas (33%), in double mutant TGR5−/−;Pkhd1del2/del2 mice (both males and females). Supporting Table 4 and Figure 5A. The mouse serum biochemistries in each group were comparable (Supporting Table 3). Reduced quantity of PCNA-positive nuclei by three times was linked to attenuated hepatic cystogenesis in double mutant TGR5−/−;Pkhd1del2/del2 mice (Fig. 5B).
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| Cell Assay |
Cell Line: shRNA-transfected ADPKD cholangiocytes
Concentration: 100, 200 µM Incubation Time: 24 hours Result: Inhibited proliferation (by 32-48%) triggered by pre-treatment of cystic cholangiocytes with TLCA. Cell Proliferation Assay: Control and ADPKD human cystic cholangiocytes were plated at 2500 cells/well and grown for 24-48 hours. Cells were then treated with SBI-115 (100 or 200 µM), pasireotide (20 µM), or a combination, for an additional 24 hours. Cell proliferation was assessed using the CellTiter 96 Aqueous One Solution Cell Proliferation Assay (MTS) and by direct cell counting using an automated cell counter. For TGR5 antagonism studies, cells were often pre-treated with the TGR5 agonist TLCA (25 µM) to stimulate proliferation before adding SBI-115. [1] cAMP Detection Assay: Cholangiocytes (10,000 cells/well) were incubated with SBI-115 (100 or 200 µM), pasireotide (20 µM), or a combination, for 15-30 minutes. In some experiments, cells were first stimulated with TLCA (25 µM). Intracellular cAMP levels were measured using the Bridge-It cAMP designer cAMP assay. [1] 3-D Cholangiocyte Spheroid Growth Assay: Cholangiocyte spheroids were generated using a hanging drop method. Drops containing 10,000 cholangiocytes in suspension were formed in a 96-well hanging drop plate. Spheroids formed within 24 hours and were grown for an additional 96 hours before treatment with SBI-115 (200 µM), pasireotide (20 µM), or both. Microphotographs were taken before and after treatment, and the circumference of the spheroids was measured to assess growth. [1] |
| References | |
| Additional Infomation |
SBI-115 (m-Tolyl-5-chloro-2-(ethanesulfonyl)pyrimidine-4-carboxylic acid ester) is a novel small molecule TGR5 antagonist. [1] It was discovered by high-throughput screening of a library of 50,000 compounds using CHO-K1 cells expressing TGR5. [1] The compound was found to be >95% pure by HPLC. [1] This study suggests that SBI-115 may be a novel potential therapy for polycystic liver disease (PLD) by inhibiting TGR5 and reducing cAMP levels in cystic bile duct cells. [1] Data suggest that in PLD, the simultaneous use of a TGR5 antagonist (SBI-115) and a somatostatin receptor agonist (paretide) to target elevated cAMP levels may be more effective than either drug alone in inhibiting bile duct cell proliferation and cyst growth. [1]
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| Molecular Formula |
C14H13CLN2O4S
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|---|---|
| Molecular Weight |
340.782021284103
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| Exact Mass |
340.03
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| Elemental Analysis |
C, 49.34; H, 3.85; Cl, 10.40; N, 8.22; O, 18.78; S, 9.41
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| CAS # |
882366-16-7
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| PubChem CID |
18879973
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| Appearance |
White to off-white solid powder
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| LogP |
2.9
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
22
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| Complexity |
494
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
IJPXOPBVXVPPEW-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C14H13ClN2O4S/c1-3-22(19,20)14-16-8-11(15)12(17-14)13(18)21-10-6-4-5-9(2)7-10/h4-8H,3H2,1-2H3
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| Chemical Name |
(3-methylphenyl) 5-chloro-2-ethylsulfonylpyrimidine-4-carboxylate
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| Synonyms |
SBI115; SBI 115; SBI-115
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO: 68~150 mg/mL (199.5~440.2 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 3 mg/mL (8.80 mM) in 2% DMSO + 40% PEG300 + 5% Tween80 + 53% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.34 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 25.0 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.34 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 5 mg/mL (14.67 mM) in Corn Oil (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.9344 mL | 14.6722 mL | 29.3444 mL | |
| 5 mM | 0.5869 mL | 2.9344 mL | 5.8689 mL | |
| 10 mM | 0.2934 mL | 1.4672 mL | 2.9344 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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