| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 50mg |
|
||
| 100mg | |||
| Other Sizes |
Purity: ≥98%
| Targets |
Transglutaminase 2 (TG2) (no IC50/Ki/EC50 values reported) [2]
KCC009 targets tissue transglutaminase 2 (TG2), an enzyme involved in post-translational modification of proteins and cell survival signaling. It is a competitive TG2 inhibitor with an IC50 of 20 μM. |
|---|---|
| ln Vitro |
The inhibition rate of H1299/WT-p53 cells is 15.33±1.46 (%) and the inhibition rate of H1299/M175H-p53 cells is 14.31±1.90 (%) at a dose of 3.91 uM of KCC009 [1].
KCC009 (1.0 mM for 24 h) disrupted the assembly of linear strands of fibronectin in the ECM of U87MG glioblastoma cells as shown by immunofluorescent staining, resulting in small clumps of fibronectin in the cytosol with minimal presence in the ECM. TG2 stained diffusely throughout the cytosol. [2] In vitro, KCC009 disrupts fibronectin assembly in the extracellular matrix and sensitizes cancer cells to chemotherapy by blocking TG2-mediated survival signaling. It disrupts the adhesion plaque complex on the surface of glioblastoma cells, resulting in a decrease in cell migration ability. |
| ln Vivo |
In an orthotopic DBT-FG glioblastoma mouse model, KCC009 (35 mg/kg/day i.p. for 5 days) disrupted fibronectin assembly in the ECM, with diminished strands of fibronectin compared to vehicle-treated tumors. Daily intraperitoneal injections of KCC009 at 50 mg/kg significantly suppressed tumor-associated TG2 activity at 15 min after the last dose (1.2 ± 0.3 vs 2.1 ± 0.2 nmol/h/mg protein in BCNU alone, P<0.02), with full recovery by 24 h (2.1 ± 0.2 vs 2.2 ± 0.3). [2]
In mice bearing orthotopic DBT-FG glioblastomas, KCC009 (50 mg/kg i.p. daily) combined with BCNU (5 mg/kg on day 10, then 2.5 mg/kg every 5 days) significantly decreased relative bioluminescence on days 18 and 21 compared to vehicle, KCC009 alone, or BCNU alone (ANOVA P≤0.004). The combination increased apoptosis as measured by TUNEL staining: 3.2% in combination vs 1.0% in vehicle, 1.5% in KCC009 alone, and 1.4% in BCNU alone (P<0.002 vs control, P<0.01 vs KCC009 alone, P<0.02 vs BCNU alone). KCC009 plus BCNU significantly prolonged median survival from 29 days (control) to 37 days (P<0.0004, log rank). Monotherapy with either agent showed a trend toward increased survival but did not reach statistical significance. [2] In vivo, KCC009 sensitizes orthotopic glioblastomas to chemotherapy. It exhibits potent radiosensitization effects in human lung cancer cells expressing wild-type or mutant p53 with different mechanisms. It induces p53 pathway-dependent sensitization. |
| Enzyme Assay |
TG2 activity was measured using a putrescine incorporation assay. Frozen human tissue samples (non-neoplastic brain, anaplastic astrocytomas, glioblastomas) or tumor samples from mice were assayed. Activity was expressed as pmoles/min/mg protein or nmol/h/mg protein. In mice with intracranial DBT tumors, TG2 activity was measured 15 min or 24 h after the last dose of KCC009 (50 mg/kg i.p. daily) or vehicle. [2]
Cell-free TG2 inhibition assays are performed using purified TG2 enzyme and a substrate such as monodansylcadaverine or biotinylated peptides. KCC009 is incubated with the enzyme and substrate, and the incorporation of the fluorescent or biotinylated probe into the substrate is measured to determine the IC50. |
| Cell Assay |
U87MG glioblastoma cells were treated with vehicle or 1.0 mM KCC009 for 24 h. Immunofluorescent staining was performed for fibronectin (red), TG2 (green), and nuclei (blue). Merged images showed colocalization (yellow-orange). KCC009 treatment prevented assembly of fibronectin strands along the cell surface. [2]
TG2 downregulation was achieved using lentiviral RNAi (Sigma) targeting human TG2. U87MG cells were infected with lentiviral particles for 24 h, selected with puromycin, and clones were examined by Western blot for TG2 expression. Immunofluorescent staining of clones with reduced TG2 showed decreased fibronectin assembly in the ECM similar to KCC009-treated cells. [2] In vitro cellular assays are performed using cancer cell lines such as glioblastoma or lung cancer cells. Cells are treated with KCC009 alone or in combination with chemotherapy or radiation. Cell viability, migration, and apoptosis are measured to assess its chemosensitizing and radiosensitizing effects. |
| Animal Protocol |
Orthotopic DBT-FG glioblastoma model: Balb/C mice (female, 8 weeks old, ~20 g) were stereotactically injected with 10,000 DBT-FG cells (transduced with luciferase/GFP) in the right frontal lobe. For in vivo efficacy, mice were divided into four groups (n=10 per group for survival, n=5 for imaging): vehicle-only, KCC009-only, BCNU-only, and KCC009+BCNU combination. KCC009 (50 mg/kg) was administered intraperitoneally daily starting on day 7 after tumor implantation. BCNU was given intraperitoneally at 5 mg/kg on day 10, then 2.5 mg/kg every 5 days thereafter. For TG2 activity measurement, mice received daily i.p. injections of KCC009 50 mg/kg or vehicle, and were sacrificed 15 min or 24 h after the last dose. For fibronectin assembly studies, mice were treated with KCC009 35 mg/kg i.p. daily for 5 days. Bioluminescence imaging was performed after i.p. injection of D-luciferin (150 µg/g) using a CCD camera system on days 7, 10, 14, 18, and 21. [2]
In vivo efficacy is evaluated in orthotopic glioblastoma models or xenograft models of lung cancer. KCC009 is administered alone or in combination with chemotherapy or radiation. Tumor growth is monitored, and tissues are collected for histopathological examination and analysis of TG2 inhibition and apoptosis markers. |
| ADME/Pharmacokinetics |
KCC009 (C21H22BrN3O5) has a molecular weight of 476.32 g/mol. Its chemical name is benzyl N-[(2S)-1-[(3-bromo-4,5-dihydro-1,2-oxazol-5-yl)methylamino]-3-(4-hydroxyphenyl)-1-oxopropan-2-yl]carbamate. It is an irreversible active-site inhibitor of TG2.
|
| Toxicity/Toxicokinetics |
No drug-related fatality or serious adverse effect was observed in any of the KCC009-treated cohorts. Doses up to 50 mg/kg/day were well tolerated in mice, and repeat daily doses up to 100 mg/kg have been well tolerated in mice and rats (data not shown). [2]
The toxicity of KCC009 is related to its inhibition of TG2. As a sensitizer to chemotherapy and radiation, its toxicity profile may be influenced by the combination with other treatments. Preclinical studies would have evaluated its safety profile, including potential effects on wound healing and tissue remodeling. |
| References |
|
| Additional Infomation |
KCC009 is a selective small molecule irreversible inhibitor of transglutaminase 2 (TG2). It disrupts the assembly of fibronectin in the ECM by inhibiting TG2 activity, thereby interfering with cell-matrix interactions that promote survival and chemoresistance. In glioblastomas, TG2 activity is elevated compared to non-neoplastic brain. KCC009 crosses the blood-tumor barrier in the CNS and sensitizes tumors to BCNU chemotherapy. The mechanism involves blocking the permissive remodeling of fibronectin, which is associated with resistance to apoptosis. [2]
KCC009 is a potent and selective transglutaminase 2 inhibitor. It exhibits potent radiosensitization effects in human lung cancer cells expressing wild-type or mutant p53 with different mechanisms. It disrupts fibronectin assembly in the extracellular matrix and sensitizes orthotopic glioblastomas to chemotherapy. It is also known as KCC-009 and KCC 009. |
| Exact Mass |
475.074
|
|---|---|
| CAS # |
744198-19-4
|
| Related CAS # |
120244-89-5 (R-isomer);744198-19-4 (KCC009);
|
| PubChem CID |
644312
|
| Appearance |
Off-white to light yellow solid powder
|
| LogP |
3.323
|
| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
30
|
| Complexity |
603
|
| Defined Atom Stereocenter Count |
1
|
| SMILES |
OC=1C=CC(=CC1)C[C@@H](C(NCC2CC(Br)=NO2)=O)NC(OCC3=CC=CC=C3)=O
|
| InChi Key |
MRULUIQNANUWTK-ZVAWYAOSSA-N
|
| InChi Code |
InChI=1S/C21H22BrN3O5/c22-19-11-17(30-25-19)12-23-20(27)18(10-14-6-8-16(26)9-7-14)24-21(28)29-13-15-4-2-1-3-5-15/h1-9,17-18,26H,10-13H2,(H,23,27)(H,24,28)/t17?,18-/m0/s1
|
| Chemical Name |
benzyl ((2S)-1-(((3-bromo-4,5-dihydroisoxazol-5-yl)methyl)amino)-3-(4-hydroxyphenyl)-1-oxopropan-2-yl)carbamate
|
| Synonyms |
KCC009 KCC-009 KCC 009.
|
| 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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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) |
DMSO : ~250 mg/mL (~524.86 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.37 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 20.8 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: ≥ 2.08 mg/mL (4.37 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 20.8 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: ≥ 2.08 mg/mL (4.37 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
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
