| Size | Price | Stock | Qty |
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| 10mg |
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Purity: ≥98%
JK184 is a novel and potent Hedgehog (Hh) pathway inhibitor with IC50 of 30 nM in mammalian cells. JK184 can specially inhibit glioma (Gli)-dependent transcriptional activity in the Hedgehog (Hh) pathway in a dose dependent manner and showed great promise for cancer therapeutics. JK184 significantly inhibits proliferation of HUVECs with IC50 of 6.3 μg/mL after three days incubation. To evaluate anti-tumor effect of JK184, MTT assay is conducted in Panc-1 and BxPC-3 cells after administration with indicated concentrations of compounds, half maximal inhibitory concentration (IC50) of JK184 (23.7 ng/mL in anc-1 and 34.3 ng/mL in BxPC-3).
| Targets |
JK184 targets Gli (effector molecule of Hedgehog (Hh) signaling pathway); no IC50/Ki/EC50 values provided [1]
JK184 targets GLI1 (glioma-associated oncogene homolog 1, effector of Hedgehog pathway); IC50 values for inhibiting proliferation of EMT cell lines (HMLE-shEcad) lower than non-EMT cell lines (HMLE-shGFP) [2] |
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| ln Vitro |
JK184 is intended to counteract Hh signaling by dose-dependently suppressing transcriptional activity that is glioma (Gli)-dependent. After three days incubation, JK184 significantly inhibits HUVEC proliferation, with an IC50 of 6.3 μg/mL. After administering the indicated concentrations of compounds to Panc-1 and BxPC-3 cells, the half maximal inhibitory concentration (IC50) of JK184 is measured (23.7 ng/mL in Anc-1 and 34.3 ng/mL in BxPC-3)[1]. This assay is used to assess the anti-tumor effect of JK184. Compared to MCF10a, MTSV1-7, HMLE-shGFP, and HMLE-pBP cells, claudin-low cell lines are more susceptible to JK184 treatment. In these cell lines, JK184 caused a dose-dependent reduction in the levels of GLI1 (glioma-associated oncogene homolog 1) transcript and protein. The percentage of HMLE-shEcad cells that stained with Annexin-V but tested negative for propidium iodide (PI) (P<0.0001, t test) increases when treated with the IC50 dose of JK184[2].
1. JK184 encapsulated in MPEG-PCL micelles shows increased cytotoxicity and cellular uptake compared with free JK184 in Panc-1 and BxPC-3 pancreatic cancer cells; micellar JK184 induces more apoptosis and blocks cell proliferation in these cell lines [1] 2. JK184 micelles exhibit sustained in vitro release behavior and stronger inhibitory effect on proliferation, migration and invasion of HUVECs than free JK184 [1] 3. JK184 (0.002/0.004 μM, 72 h treatment) downregulates GLI1 protein and mRNA levels in HMLE-shEcad (EMT) cells (immunoblot and real-time PCR, P≤0.05, P≤0.005) [2] 4. JK184 (0.002/0.004 μM, 72 h) reduces GLI1 transcript levels in claudin-low breast cancer cell lines (MDA.MB.157, MDA.MB.436, BT549) (real-time RT-PCR, P≤0.05, P≤0.005) [2] 5. JK184 (0.02 μM, 4 days) induces apoptosis in claudin-low breast cancer cell lines (Annexin-V/PI flow cytometry) [2] 6. JK184 is more effective at inhibiting proliferation of EMT (HMLE-shEcad, HMLE-Snail) cells than non-EMT (HMLE-shGFP, HMLE-pBP) cells (dose-response curve analysis) [2] 7. JK184 (1 μM, 16 h) downregulates GLI1 transcript levels in HMLE-shEcad cells (real-time PCR, P<0.05, P<0.005) [2] |
| ln Vivo |
JK184 (5 mg/kg, intravenously injected) is a promising candidate for an antitumor drug that targets Hh signaling because it shows good anti-proliferative activity in subcutaneous Panc-1 and BxPC-3 tumor models. JK184, however, has a subpar bioavailability and pharmacokinetic profile [1].
1. JK184 micelles show stronger tumor growth inhibition than free JK184 in subcutaneous Panc-1 and BxPC-3 pancreatic tumor models in nude mice; histological analysis reveals more apoptosis, decreased microvessel density, and reduced expression of CD31, Ki67, and VEGF in tumor tissues; micellar JK184 more potently inhibits Gli expression in Hh signaling pathway in tumor tissues [1] 2. No direct in vivo data of free JK184 on breast cancer models; GLI1 knockdown (mimicking JK184 effect) reduces tumor growth of orthotopic MDA.MB.436 (claudin-low breast cancer) xenografts in mice (n=6 animals, reduced tumor volume over time and lower final tumor weight at 7 weeks post-injection; replicate study with n=4 animals confirms reduced tumor weight) [2] |
| Cell Assay |
1. Cytotoxicity/uptake assay (Panc-1/BxPC-3): Panc-1 and BxPC-3 cells were treated with free JK184 or JK184 micelles; cell viability was assessed to measure cytotoxicity, and cellular uptake of the drug was quantified to compare free vs micellar formulations [1]
2. Apoptosis/proliferation assay (Panc-1/BxPC-3): Cells were treated with free JK184 or JK184 micelles; apoptosis was detected by relevant assays (not specified), and cell proliferation was evaluated by measuring cell number/viability over time to compare the effects of free and micellar drug [1] 3. HUVEC functional assay: HUVECs were treated with free JK184 or JK184 micelles; cell proliferation was assessed by viability assays, migration by wound healing/transwell assay, and invasion by matrigel transwell assay to compare inhibitory effects [1] 4. Proliferation assay (EMT/claudin-low breast cancer cells): HMLE-shEcad, HMLE-shGFP, HMLE-Snail, HMLE-pBP (EMT/non-EMT) and claudin-low cell lines (MDA.MB.157, MDA.MB.436, BT549) were seeded in 96-well plates and treated with serial concentrations of JK184; cell proliferation was measured to generate dose-response curves [2] 5. GLI1 expression assay (HMLE/claudin-low cells): HMLE-shEcad and claudin-low cell lines were treated with 0.002/0.004 μM JK184 for 72 h or 1 μM JK184 for 16 h; total protein was extracted for immunoblotting to detect GLI1 levels (GAPDH as loading control), and total RNA was extracted for real-time RT-PCR to quantify GLI1 mRNA levels [2] 6. Apoptosis assay (claudin-low cells): Claudin-low breast cancer cells were treated with 0.02 μM JK184 for 4 days; cells were stained with Annexin-V and propidium iodide (PI), and apoptosis was analyzed by flow cytometry [2] 7. Migration/clonogenicity assay (claudin-low cells): BT549/MDA.MB.436 cells were treated with JK184 (or GLI1 shRNA as mimic); migration was assessed by Boyden chamber assay (cells plated in top chamber with 1% FBS, lower chamber with 10% FBS, migrated cells stained/counted after 16 h), and clonogenicity by colony formation assay (cells plated in soft agar, colonies counted after 2–3 weeks) [2] |
| Animal Protocol |
5 mg/kg, injected intravenously
Mice with Panc-1 and BxPC-3 tumor models 1. Pancreatic tumor xenograft assay (nude mice): Nude mice were subcutaneously inoculated with Panc-1 or BxPC-3 cells to establish tumor models; after tumors became palpable, mice were randomized into groups and treated with free JK184 or JK184 micelles (administration route/frequency/dissolution formula not specified); tumor volume was measured regularly, and at the end of the study, tumors were harvested for histological analysis (H&E staining, immunohistochemistry for CD31, Ki67, VEGF, Gli) [1] 2. Breast cancer orthotopic xenograft assay (mice): MDA.MB.436 cells (claudin-low breast cancer) were infected with non-targeting shRNA or GLI1 shRNA (mimicking JK184 effect); cells were orthotopically injected into mammary fat pads of mice; tumor volume was measured over time, and mice were euthanized at 7 weeks post-injection to weigh tumors (replicate study with independent cell injection and tumor weight measurement) [2] |
| ADME/Pharmacokinetics |
Compared to free JK184, JK184 encapsulated in MPEG-PCL micelles has a longer circulation time in the blood; however, specific parameters (half-life, oral bioavailability, absorption/distribution/metabolism/excretion) are not provided [1].
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| References |
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| Additional Infomation |
1. JK184 is a specific inhibitor of Gli in the Hedgehog (Hh) pathway and has the potential for cancer treatment. JK184 MPEG-PCL micelles were prepared by solid-phase dispersion method without the need for surfactants or toxic organic solvents to develop aqueous formulations and improve antitumor activity[1]. 2. JK184 targets GLI1, an effector molecule of the Hh pathway. GLI1 is highly expressed in claudin-low breast cancer (triple-negative, poor prognosis) and EMT cell lines. GLI1 is essential for maintaining the stem cell-like/mesenchymal characteristics (viability, motility, clonogenic ability, self-renewal ability) of claudin-low breast cancer cells. JK184 acts on non-canonical GLI1 activation in claudin-low/EMT cells, and GLI1 interacts with the NFκB pathway (p65 binds to the GLI1 promoter)[2].
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| Molecular Formula |
C19H18N4OS
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|---|---|---|
| Molecular Weight |
350.44
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| Exact Mass |
350.12
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| CAS # |
315703-52-7
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| Related CAS # |
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| PubChem CID |
1069686
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| Appearance |
Off-white to pink solid powder
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| Density |
1.3±0.1 g/cm3
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| Index of Refraction |
1.683
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| LogP |
4.04
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
25
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| Complexity |
432
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
ROYXIPOUVGDTAO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H18N4OS/c1-3-24-15-9-7-14(8-10-15)21-19-22-16(12-25-19)18-13(2)20-17-6-4-5-11-23(17)18/h4-12H,3H2,1-2H3,(H,21,22)
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| Chemical Name |
N-(4-ethoxyphenyl)-4-(2-methylimidazo[1,2-a]pyridin-3-yl)-1,3-thiazol-2-amine
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| Synonyms |
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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 |
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| 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) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.13 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.13 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.13 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.8536 mL | 14.2678 mL | 28.5356 mL | |
| 5 mM | 0.5707 mL | 2.8536 mL | 5.7071 mL | |
| 10 mM | 0.2854 mL | 1.4268 mL | 2.8536 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.
JK184 is more effective at inhibiting EMT cell proliferation.Breast Cancer Res.2014 Sep 25;16(5):444. |
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JK184 inhibits growth of claudin-low cell lines.Breast Cancer Res.2014 Sep 25;16(5):444. |
Decrease inGLI1expression inhibits cell migration and anchorage-independent growth.Breast Cancer Res.2014 Sep 25;16(5):444. |
EMT and claudin-low cells are insensitive to Hedgehog (Hh) pathway inhibitors.Breast Cancer Res.2014 Sep 25;16(5):444. |
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Crosstalk between NFκB and GLI1 signaling pathways.Breast Cancer Res.2014 Sep 25;16(5):444. |
Claudin-low cell lines express higher transcript and protein levels ofGLI1.Breast Cancer Res.2014 Sep 25;16(5):444. |
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