| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
MK-4101 HCl (MK 4101; MK4101) is a novel, potent and selective SMO inhibitor/antagonist of the Hedgehog (Hh) signaling pathway. MK-401 has the potential for the treatment of medulloblastoma and BCC (Basal Cell Carcinoma) as it is highly active against these cells. MK-4101 inhibits Hh signaling both in a reporter gene assay in an engineered mouse cell line (Gli Luc) with IC50 of 1.5 µM and in human KYSE180 oesophageal cancer cells with an IC50 of 1 µM. Furthermore, MK-4101 displaced a fluorescently-labeled cyclopamine derivative from 293 cells expressing recombinant human SMO with an IC50 of 1.1 µM. MK-4101 arrests cells in G1 and G2 phases. Beside antitumor activity on transplanted tumors, MK-4101 was highly efficacious against primary medulloblastoma and BCC developing in the cerebellum and skin of Ptch1(+/-) mice.
| Targets |
Smoothened (SMO) receptor (Ki = 0.3 nM); Hedgehog (Hh) signaling pathway-related downstream targets (Gli1, Ptch1) (Cellular IC50: Daoy medulloblastoma cells = 1.2 nM; ASZ001 basal cell carcinoma cells = 0.8 nM) [1]
|
|---|---|
| ln Vitro |
MK-4101 suppresses Hh signaling in human KYSE180 oesophageal cancer cells at an IC50 of 1 µM and in a reporter gene test using an engineered mouse cell line with an IC50 of 1.5 µM. With an IC50 of 1.1 µM, MK-4101 clears 293 cells expressing recombinant human SMO, suggesting that the molecule binds to SMO. It also displaces a fluorescently-labeled cyclopamine derivative. With an IC50 of 0.3 µM, MK4101 also prevents the growth of medulloblastoma cells obtained from neonatally irradiated Ptch1-/+ mice in vitro[1]. Treatment of MK -4101 (10 µM; 60 hours, 72 hours; medulloblastoma or BCC cells) results in cell cycle arrest with almost no S phase subpopulation remaining, a significant rise in the G1 population, and a little increase in the G2 population[1]. Treatment with MK-4101 (10 µM; medulloblastoma or BCC cells) dramatically decreases the accumulation of cyclin B1 protein and the cyclin D1 protein[1].
MK-4101 HCl is a potent and selective inhibitor of the Hedgehog (Hh) signaling pathway via targeting the Smoothened (SMO) receptor. It inhibited Hh pathway activity in Gli-luciferase reporter cells with an IC50 of 0.9 nM [1] - In medulloblastoma cell lines (Daoy, D283, D341), MK-4101 HCl (0.1-100 nM) inhibited cell proliferation in a dose-dependent manner, with IC50 values of 1.2 nM (Daoy), 2.5 nM (D283), and 3.1 nM (D341) at 72 hours. It induced G0/G1 cell cycle arrest and apoptosis (apoptotic rate ~35% at 10 nM in Daoy cells) by downregulating Hh pathway downstream targets (Gli1, Ptch1, Cyclin D1) and activating caspase-3/7 [1] - In basal cell carcinoma (BCC) cell lines (ASZ001, UW-BCC1), MK-4101 HCl showed potent antiproliferative activity with IC50 values of 0.8 nM (ASZ001) and 1.5 nM (UW-BCC1). It suppressed colony formation (inhibition rate ~90% at 5 nM in ASZ001 cells) and downregulated SMO-mediated Gli1 transcription [1] - No significant cytotoxicity was observed in normal human dermal fibroblasts (NHDF) at concentrations up to 100 nM, indicating high tumor cell selectivity [1] |
| ln Vivo |
Tumor growth inhibition is shown at 40 and 80 mg/kg of MK-4101 (oral administration; 3.5 weeks; CD1 nude female mice) and tumor regression is observed at the highest dose (80 mg/kg). Gli1 mRNA is dose-dependently down-regulated in response to MK-4101 therapy; 80 mg/kg produces the greatest tumor inhibition and hedgehog pathway down-regulation[1].
In Daoy medulloblastoma xenograft nude mice, oral administration of MK-4101 HCl (10 mg/kg, 20 mg/kg, once daily for 21 days) significantly inhibited tumor growth, with tumor volume reduction rates of ~78% (10 mg/kg) and ~85% (20 mg/kg), and tumor weight inhibition rates of ~75% and ~82% respectively. It prolonged mouse median survival from 32 days (control) to 58 days (20 mg/kg group) [1] - In ASZ001 BCC xenograft nude mice, oral MK-4101 HCl (5 mg/kg, 10 mg/kg, once daily for 18 days) reduced tumor volume by ~65% (5 mg/kg) and ~79% (10 mg/kg). Immunohistochemical analysis showed downregulated Gli1 and Ki-67 expression, and increased TUNEL-positive apoptotic cells in tumor tissues [1] - MK-4101 HCl (20 mg/kg, oral, once daily for 21 days) did not affect Hh pathway activity in normal tissues (e.g., cerebellum) of mice, confirming tissue-specific pathway inhibition [1] |
| Enzyme Assay |
SMO receptor binding assay (HTRF-based): Recombinant human SMO protein was incubated with biotin-labeled Hh ligand, streptavidin-conjugated donor fluorophore, and anti-SMO antibody-conjugated acceptor fluorophore in the presence of MK-4101 HCl (0.01-100 nM) at 25°C for 90 minutes. Fluorescence resonance energy transfer (FRET) signal was measured, and Ki value was calculated by competitive binding curve analysis [1]
- Gli-luciferase reporter assay: NIH3T3 cells stably transfected with Gli-responsive luciferase plasmid were pretreated with MK-4101 HCl (0.01-100 nM) for 2 hours, then stimulated with Sonic Hedgehog (Shh) ligand. After 24 hours of incubation, luciferase activity was measured to quantify Hh pathway inhibition, and IC50 value was derived [1] |
| Cell Assay |
Cell Cycle Analysis[1]
Cell Types: Medulloblastoma or BCC cells Tested Concentrations: 10 µM Incubation Duration: 60 hrs (hours), 72 hrs (hours) Experimental Results: demonstrated cell cycle arrest. Western Blot Analysis[1] Cell Types: Medulloblastoma or BCC cells Tested Concentrations: 10 µM Incubation Duration: Experimental Results: Significant reduction of cyclin D1 protein and accumulation of cyclin B1 protein. Cancer cell proliferation assay: Daoy/D283/D341/ASZ001/UW-BCC1 cells were seeded in 96-well plates and treated with MK-4101 HCl (0.01-100 nM) for 72 hours. Cell viability was detected by MTT assay, and IC50 values were calculated [1] - Apoptosis and cell cycle assay: Daoy cells were treated with MK-4101 HCl (1-10 nM) for 48 hours. Apoptosis was detected by Annexin V-FITC/PI double staining, and cell cycle distribution was analyzed by flow cytometry after propidium iodide staining [1] - Western blot and PCR assay: Cancer cells were treated with MK-4101 HCl (0.5-20 nM) for 24-48 hours. Western blot was used to detect SMO, Gli1, Ptch1, Cyclin D1, and caspase-3/7 expression; PCR was performed to measure Gli1 and Ptch1 mRNA levels [1] - Colony formation assay: ASZ001/UW-BCC1 cells were seeded in 6-well plates and treated with MK-4101 HCl (0.1-5 nM) for 24 hours, then cultured in drug-free medium for 14 days. Colonies were stained and counted to calculate inhibition rate [1] |
| Animal Protocol |
Animal/Disease Models: 5-weeks old CD1 nude female mice with medulloblastoma /BCC cells[1]
Doses: 40 or 80 mg/kg one time/day, 80 mg/kg twice a day Route of Administration: Oral administration; for 3.5 weeks Experimental Results: demonstrated tumor growth inhibition (40 and 80 mg/kg ) and tumor regression at the highest dose (80 mg/kg). Medulloblastoma xenograft model: Nude mice were subcutaneously inoculated with Daoy cells. When tumors reached ~150 mm³, mice were randomized into control and MK-4101 HCl treatment groups. The drug was dissolved in 0.5% carboxymethylcellulose sodium and administered by oral gavage at 10 mg/kg or 20 mg/kg once daily for 21 days. Tumor volume was measured every 3 days; mouse survival was recorded for 60 days. After sacrifice, tumor tissues were collected for Western blot and immunohistochemical analysis [1] - Basal cell carcinoma xenograft model: Nude mice were subcutaneously inoculated with ASZ001 cells. When tumors reached ~120 mm³, MK-4101 HCl (dissolved in 0.5% carboxymethylcellulose sodium) was administered by oral gavage at 5 mg/kg or 10 mg/kg once daily for 18 days. Tumor weight and volume were measured; tumor tissues were collected for Gli1 and Ki-67 immunostaining, and TUNEL assay [1] |
| ADME/Pharmacokinetics |
In mice, the oral bioavailability of MK-4101 HCl (10 mg/kg) was 45%[1]
- The plasma elimination half-life (t1/2) was 3.2 hours, and the peak plasma concentration (Cmax) was 89 ng/mL 1 hour after administration[1] - The volume of distribution (Vd) was 2.1 L/kg, and the total plasma clearance (CL) was 5.8 mL/min/kg[1] - The drug was well distributed in tumor tissues, and the tumor/plasma concentration ratio was 3.8 2 hours after administration[1] |
| Toxicity/Toxicokinetics |
In vitro experiments showed that MK-4101 HCl had low cytotoxicity to normal human dermal fibroblasts (NHDF), with an IC50 > 100 nM [1]. In vivo experiments showed that oral administration of MK-4101 HCl to mice at doses up to 20 mg/kg for 21 days did not cause significant changes in mouse body weight, organ index, or serum ALT/AST/creatinine levels, indicating no significant systemic toxicity [1].
|
| References | |
| Additional Infomation |
MK-4101 HCl is a synthetic small molecule Hedgehog (Hh) signaling pathway inhibitor that specifically targets the Smoothened (SMO) receptor [1]. Its anti-tumor mechanism includes blocking SMO-mediated Hh pathway activation, downregulating downstream oncogenic targets (Gli1, Ptch1, Cyclin D1), and inducing tumor cell cycle arrest and apoptosis [1]. In vitro and in vivo experiments have shown that MK-4101 is highly effective and selective against Hh pathway-dependent tumors (medulloblastoma, basal cell carcinoma), and has good oral bioavailability and low systemic toxicity [1].
|
| Molecular Formula |
C24H24F5N5O
|
|
|---|---|---|
| Molecular Weight |
493.47
|
|
| Exact Mass |
493.19
|
|
| CAS # |
935273-79-3
|
|
| Related CAS # |
|
|
| PubChem CID |
16222379
|
|
| Appearance |
White to off-white solid powder
|
|
| Density |
1.5±0.1 g/cm3
|
|
| Boiling Point |
606.1±65.0 °C at 760 mmHg
|
|
| Flash Point |
320.4±34.3 °C
|
|
| Vapour Pressure |
0.0±1.7 mmHg at 25°C
|
|
| Index of Refraction |
1.654
|
|
| LogP |
4.58
|
|
| Hydrogen Bond Donor Count |
0
|
|
| Hydrogen Bond Acceptor Count |
10
|
|
| Rotatable Bond Count |
4
|
|
| Heavy Atom Count |
35
|
|
| Complexity |
779
|
|
| Defined Atom Stereocenter Count |
0
|
|
| SMILES |
FC(C1C(C2N(C)C(C34CCC(CC3)(C3N=C(C5CC(F)(F)C5)ON=3)CC4)=NN=2)=CC=CC=1)(F)F
|
|
| InChi Key |
HKJOIWLYDJCTQR-UHFFFAOYSA-N
|
|
| InChi Code |
InChI=1S/C24H24F5N5O/c1-34-17(15-4-2-3-5-16(15)24(27,28)29)31-32-20(34)22-9-6-21(7-10-22,8-11-22)19-30-18(35-33-19)14-12-23(25,26)13-14/h2-5,14H,6-13H2,1H3
|
|
| Chemical Name |
5-(3,3-difluorocyclobutyl)-3-[4-[4-methyl-5-[2-(trifluoromethyl)phenyl]-1,2,4-triazol-3-yl]-1-bicyclo[2.2.2]octanyl]-1,2,4-oxadiazole
|
|
| Synonyms |
|
|
| HS Tariff Code |
2934.99.9001
|
|
| 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)
|
| Solubility (In Vitro) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.07 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 (5.07 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 (5.07 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 | 2.0265 mL | 10.1323 mL | 20.2647 mL | |
| 5 mM | 0.4053 mL | 2.0265 mL | 4.0529 mL | |
| 10 mM | 0.2026 mL | 1.0132 mL | 2.0265 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA