1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEAndrographolideCat. No.: HY-N0191CAS No.: 5508-58-7Synonyms: Andrographis分式: CHO分量: 350.45作靶點(diǎn): NF-B; Autophagy作通路: NF-B; Autophagy儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 50 mg/mL (1

2、42.67 mM; Need ultrasonic)H2O : 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (7.13 mM); Clear solution2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (7.13 mM); Clear solution1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubili

3、ty: 2.5 mg/mL (7.13 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 Andrographolide種 NF-B 抑制劑，通過共價(jià)修飾內(nèi)細(xì)胞中 p50 的半胱氨酸殘 抑制 NF-B 活化，不影響 IB 降解或 p50/p65 核易位。IC50 & Target p50體外研究 Andrographolide (AP) concentration-dependently suppresses receptor activator of nuclear factor kappa Bligand (RANKL)-mediated osteocla

4、st differentiation and bone resorption in vitro and reduces the expressionof osteoclast-specific markers. Andrographolide attenuates inflammation by inhibition of TNF-induced NF-B activation through covalent modification of reduced Cys62 of p50, without affecting IB degradation orp50/p65 nuclear tra

5、nslocation. Andrographolide also inhibits the ERK/MAPK signalling pathway withoutaffecting p38 or JNK signalling. Andrographolide inhibits osteoclast differentiation of RAW 264.7 cells in aconcentration-dependent manner. Andrographolide suppresses osteoclast formation in a concentration-dependent ma

6、nner without any obvious cytotoxic effects, in both BMMs and RAW 264.7 cells.Andrographolide treatment substantially reduces the area of bone resorption. Only approximately 30% of thebone resorption observed in the control group is achieved after treatment with 2.5M Andrographolide.Osteoclastic bone

7、 resorption is almost completely inhibited after treatment with 10M Andrographolide 1.體內(nèi)研究 Treatment with Andrographolide (5 or 30mg/kg) reduces the extent of bone loss induced by LPS. Moreover,Andrographolide slightly increases the BMD and cortex thickness compared to LPS treatment. Histologicalexa

8、mination confirms the protective effects of Andrographolide on LPS-induced bone loss. LPS injectionleads to inflammatory bone erosion and increased numbers of TRAP-positive osteoclasts 1.PROTOCOLKinase Assay 1 In vitro osteoclastogenesis assays are preformed to examine the effects of Andrographolide

9、 on osteoclastdifferentiation. Bone marrow macrophages (BMM) cells are prepared. Briefly, cells extracted from the femurand tibiae of a 6-week-old C57/BL6 mouse are incubated in complete cell culture media and 30ng/mLM-CSF in a T-75cm2 flask for proliferation. When changing the medium, the cells are

10、 washed in order todeplete residual stromal cells. After reaching 90% confluence, cells are washed with PBS three times andtrypsinized for 30min to harvest BMMs. Cells adhering to the bottom of the dish are classified as BMMs;these BMMs are plated in 96-well plates at a density of 8103 cells per wel

11、l in triplicate and incubated in ahumidified incubator containing 5% CO2 at 37C for 24h. The cells are then treated with variousconcentrations of Andrographolide (0, 2.5, 5, or 10M) plus M-CSF (30ng/mL) and RANKL (50ng/mL). After5 days, cells are fixed and stained for tartrate-resistant acid phospha

12、tase (TRAP) activity. TRAP-positivemultinucleated cells with more than five nuclei are counted as osteoclasts 1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemECell Assay 1 Effects of Andrographolide on c

13、ell proliferation are determined with a CCK-8. BMMs are plated in 96-wellplates at a density of 3103 cells per well in triplicate. Twenty-four hours later, the cells are treated withincreasing concentrations of Andrographolide (0, 2.5, 5, 10 or 20M) for 2 days. Next, 10L CCK-8 is addedto each well,

14、and the plates are then incubated at 37C for an additional 2h. The optical density (OD) is thenmeasured with an ELX800 absorbance microplate reader at a wavelength of 450nm (650nm reference). Thecell viability is calculated 1.MCE has not independently confirmed the accuracy of these methods. They ar

15、e for reference only.Animal Mice 1Administration 1 C57BL/6 mice (8 weeks old) are divided into four groups of seven mice each. Mice are injected i.p. withAndrographolide (5 or 30mg/kg body weight) or PBS as a control 1 day before injection of LPS (5g/g bodyweight). Andrographolide or PBS is injected

16、 intraperitoneally every other day for 8 days. LPS is injectedintraperitoneally on days one and four. All mice are killed 8 days after the initial LPS injection, and the leftfemurs of all animals are scanned with a high-resolution micro-CT at a resolution of 9m.MCE has not independently confirmed th

17、e accuracy of these methods. They are for reference only.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) J Cell Mol Med. 2019 Jun 26. Front Microbiol. 2018 Oct 8;9:2407.See more customer validations on HYPERLINK / www.MedChemEREFERENCES1. Zhai ZJ, et al. Andrographolide suppresses RANKL-induced osteoclastogenesis in vitro and prevents inflammatory