Int 747

Manufactured by MedChemExpress

Sourced in United States

INT-747 is a small molecule compound that functions as an agonist for the farnesoid X receptor (FXR). FXR is a nuclear receptor that plays a key role in bile acid, lipid, and glucose homeostasis. INT-747 can be used as a tool compound to study the biological functions of FXR.

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Lab products found in correlation

Int 777, by MedChemExpress (4 mentions) Human cd14 microbeads, by Miltenyi Biotec (1 mentions) Mouse cd4 microbeads, by Miltenyi Biotec (1 mentions) Gm csf, by ACROBiosystems (1 mentions) Interleukin 4 (il 4), by ACROBiosystems (1 mentions) Lipopolysaccharides (lps), by Merck Group (1 mentions) Flagellin, by Thermo Fisher Scientific (1 mentions) Protease and phosphatase inhibitor, by Thermo Fisher Scientific (1 mentions) Pam3csk4, by Thermo Fisher Scientific (1 mentions) Gw4064, by Bio-Techne (1 mentions) Lipopolysaccharides (lps), by Thermo Fisher Scientific (1 mentions) Radioimmunoprecipitation assay (ripa), by Merck Group (1 mentions)

4 protocols using int 747

Modulation of BMDC and T-cell responses by bile acids

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BMDCs from the mice were extracted and cultured as previously described 26 (link). Briefly, 5×10⁵ BMDCs were stimulated using lipopolysaccharides (LPS) (1 μg/mL, Sigma-Aldrich) and bile acids (10 µM, Sigma-Aldrich) or INT-747(100 µM, MedChem Express, USA) or INT-777 (100 µM, MedChem Express) for 24 h.
Mice CD4+ T cells were isolated using mouse CD4 microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany) and cocultured with BMDCs pretreated with LCA or INT-777 at a ratio of 5:1 (CD4+ T cells: BMDCs) for 5 days.
Human CD14+ monocytes in PBMCs were isolated using Human CD14 microbeads (Miltenyi Biotec). CD14+ monocytes were incubated with 50 ng/mL IL-4 (AcroBiosystems, Newark, NJ, USA) and 100 ng/mL GM-CSF (AcroBiosystems) to induce DC maturation. 7 days later, 5×10⁵ MD-DCs were treated with 100 ng/ml LPS and 10 µM LCA or 100 µM INT-777 for 24 h.

Hu J., Zhang Y., Yi S., Wang C., Huang X., Pan S., Yang J., Yuan G., Tan S, & Li H. (2022). Lithocholic acid inhibits dendritic cell activation by reducing intracellular glutathione via TGR5 signaling. International Journal of Biological Sciences, 18(11), 4545-4559.

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Bile Acid Administration in Mice

Cited 1 time

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BAs, INT-747 and INT-777 (MedChemExpress, Monmouth Junction, NJ), were dissolved in DMSO to a stock concentration of 1 mM. For in vivo study, BAs were dissolved by adding each solvent sequentially: 10% DMSO, >90% corn oil for oral administration, and 5% DMSO >95% PBS for i.v. administration. Mice received oral gavage of 0.6-mg BAs once daily (~30 mg/kg) dissolved in a 200-μl vehicle (Hang et al., 2019 (link)), or i.v. injections of 80-μg LCA through tail vein every other day (~4 mg/kg) in 100 μl vehicle.

Shi Z., Wu X., Wu C.Y., Singh S.P., Law T., Yamada D., Huynh M., Liakos W., Yang G., Farber J.M., Wan Y.J, & Hwang S.T. (2021). Bile Acids Improve Psoriasiform Dermatitis through Inhibition of IL-17A Expression and CCL20-CCR6–Mediated Trafficking of T Cells. The Journal of investigative dermatology, 142(5), 1381-1390.e11.

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Intrathecal Drug Injection Protocol

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For single intrathecal injection, direct transcutaneous intrathecal injection was performed following previous work.²⁸, ²⁹ Briefly, a 25‐Ga 3 10 needle connected to a Hamilton syringe was inserted into the tissues between the dorsal aspects of L5 and L6 of mice, perpendicular to the vertebral column. The injection was considered successful if a tail flick was observed immediately. Then, 5 μL INT‐777, INT‐747, bicuculline SBI‐115, or Z‐guggulsterone (all the above drugs were purchased from MedChemExpress) was administered. For repetitive INT‐777 injection, the drug was injected intrathecally via a polyethylene 10 (PE‐10) tube, the tip of which was placed on the spinal lumbar enlargement level 1 week before. Whether the tube was correctly placed in the intervertebral space was confirmed by paralysis in the bilateral hind limb after a 2% lidocaine injection (3 μL) through the catheter within 30 s.

Wu Y., Qiu Y., Su M., Wang L., Gong Q, & Wei X. (2023). Activation of the bile acid receptors TGR5 and FXR in the spinal dorsal horn alleviates neuropathic pain. CNS Neuroscience & Therapeutics, 29(7), 1981-1998.

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Metabolite-modulated PGE2 secretion in cMSCs

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In stimulation experiments, cMSCs were plated in 96-well plates at 20,000 cells/well in low glucose DMEM (containing 10mM HEPES, penicillin-streptomycin, 10% FBS) and allowed to adhere overnight. The following morning, cells were fed fresh media, stimulated with ligands and incubated at 37°C for 24 hours. For TLR screen, cMSCs were individually stimulated with Pam3CSK4, LPS, flagellin, Pam2CSK4, poly IC (Invitrogen) at indicated doses or vehicle (endotoxin free water, Invitrogen). For the metabolite screen, cMSCs were stimulated with 100ng/ml of Pam3CSK4 in the presence or absence of bacterial metabolites (100 µM). For secondary validation, metabolites at 100µM, 10µM and 1µM were used. For receptor agonists, cMSCs were stimulated with 100ng/ml Pam3CSK4 in the presence or absence of GW4064 (Tocris), INT747 (MedChem Express) or INT-777 (MedChem Express). 24-hour culture supernatants were collected from the treatments and PGE2 concentration was assayed. In some experiments, cells were washed with PBS and lysed in RIPA (Sigma) buffer containing protease and phosphatase inhibitors (Thermo Fisher Scientific). These lysates were then assayed for PGE2. All the experiments utilized cMSCs between passage number 5 and 8.

Jain U., Lai C.W., Xiong S., Goodwin V.M., Lu Q., Muegge B.D., Christophi G.P., VanDussen K.L., Cummings B.P., Young E., Hambor J, & Stappenbeck T.S. (2018). Temporal regulation of the bacterial metabolite deoxycholate during colonic repair is critical for crypt regeneration. Cell host & microbe, 24(3), 353-363.e5.

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