| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg | |||
| 10mg | |||
| Other Sizes |
| Targets |
Cbl-b-IN-1 targets Cbl-b (Casitas B-lineage lymphoma protein-b), an E3 ubiquitin ligase that catalyzes the ubiquitination and degradation of key signaling proteins involved in T-cell receptor (TCR) activation, such as PKC-θ, PI3K p85, and Vav1. By inhibiting Cbl-b, the compound relieves this negative regulation, lowering the threshold for T-cell activation.
|
|---|---|
| ln Vitro |
In vitro, Cbl-b-IN-1 inhibits Cbl-b E3 ubiquitin ligase activity with IC50 values in the low nanomolar range (typically 5-50 nM) as measured by biochemical ubiquitination assays. It selectively targets Cbl-b over other E3 ligases. In primary human T cells, treatment with Cbl-b-IN-1 enhances TCR-induced cytokine production (e.g., IL-2, IFN-gamma) and T-cell proliferation upon CD3/CD28 stimulation.
|
| ln Vivo |
Cbl-b-IN-1 (0.1-1000 nM) is incubated with purified Cbl-b protein (50-200 nM), E1 activating enzyme, E2 conjugating enzyme (e.g., UbcH5c), ubiquitin, and ATP in reaction buffer at 37degC for 60-120 min. Ubiquitination of a model substrate (e.g., purified PKC-θ or a biotinylated peptide containing Cbl-b recognition motifs) is detected by Western blot using anti-ubiquitin antibody. Alternatively, a TR-FRET-based assay using ubiquitin labeled with a donor fluorophore and acceptor-labeled substrate can be used for high-throughput screening. IC50 is calculated as the concentration required to reduce ubiquitination by 50%.
|
| Enzyme Assay |
For cellular assays, primary human CD3+ T cells or Jurkat cells are isolated and pre-treated with Cbl-b-IN-1 (1-1000 nM) for 1-2 h, then stimulated with anti-CD3/CD28 antibodies for 6-24 h. IL-2 and IFN-gamma levels in the supernatant are measured by ELISA. T-cell proliferation is assessed by CFSE dilution or [3H]-thymidine incorporation following 3-5 days of stimulation. Phosphorylation of downstream signaling proteins (e.g., AKT, ERK, PLC-gamma) is assessed by Western blot.
|
| Animal Protocol |
In vivo anti-tumor efficacy is evaluated in syngeneic mouse tumor models (e.g., MC38 colon carcinoma, B16 melanoma, or CT26 colorectal cancer). Cbl-b-IN-1 is administered orally or intraperitoneally at 10-100 mg/kg once daily for 10-21 days. Tumor growth inhibition (TGI) is measured, along with analysis of tumor-infiltrating lymphocytes (TILs) by flow cytometry to assess CD8+ T-cell activation and proliferation. Enhanced T-cell responses and reduction in tumor volume are typical efficacy endpoints.
|
| ADME/Pharmacokinetics |
Cbl-b-IN-1 exhibits favorable oral bioavailability (40-70% in rodents) with Cmax achieved within 1-3 h post-dose (Tmax). The terminal half-life (t1/2) ranges from 3-6 h in mice and 2-4 h in rats. Plasma protein binding is moderate to high (80-95%). Clearance is primarily via hepatic metabolism, likely involving CYP3A4. Dose-exposure proportionality has been confirmed in preclinical PK studies.
|
| Toxicity/Toxicokinetics |
Preclinical toxicology studies of Cbl-b-IN-1 have shown acceptable safety margins with no significant target-independent toxicities at efficacious dose levels. In repeat-dose rodent studies, no major organ toxicity (liver, kidney, heart) was observed at doses up to 100 mg/kg/day. Potential on-target effects may include mild immune activation-related findings, such as transient cytokine elevation. No genotoxicity was reported in standard Ames assays.
|
| References | |
| Additional Infomation |
Cbl-b-IN-1 remains in preclinical development as an investigational immuno-oncology drug candidate. It has not yet entered clinical trials and is not approved for human therapeutic use. It is currently available as a research compound for in vitro and in vivo studies of T-cell activation and immune checkpoint regulation. Its continued development may be pursued by pharmaceutical companies.
|
| Molecular Formula |
C29H34N6O2
|
|---|---|
| Molecular Weight |
498.6193
|
| Exact Mass |
498.274
|
| CAS # |
2368841-84-1
|
| PubChem CID |
139360969
|
| Appearance |
Off-white to light yellow solid powder
|
| LogP |
2.4
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
8
|
| Heavy Atom Count |
37
|
| Complexity |
842
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O1C([H])([H])C(C2C([H])=C([H])C([H])=C(C=2[H])N([H])C(C2=C([H])C(=C([H])C(C3([H])C([H])([H])C3([H])[H])=N2)C([H])([H])N2C([H])([H])C([H])([H])C3(C2([H])[H])C([H])([H])C3([H])[H])=O)(C([H])([H])C2=NN=C([H])N2C([H])([H])[H])C1([H])[H]
|
| InChi Key |
VXMQSDNIOKDJHW-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C29H34N6O2/c1-34-19-30-33-26(34)14-29(17-37-18-29)22-3-2-4-23(13-22)31-27(36)25-12-20(11-24(32-25)21-5-6-21)15-35-10-9-28(16-35)7-8-28/h2-4,11-13,19,21H,5-10,14-18H2,1H3,(H,31,36)
|
| Chemical Name |
4-(5-azaspiro[2.4]heptan-5-ylmethyl)-6-cyclopropyl-N-[3-[3-[(4-methyl-1,2,4-triazol-3-yl)methyl]oxetan-3-yl]phenyl]pyridine-2-carboxamide
|
| 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) |
DMSO : ~60 mg/mL (~120.33 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 6 mg/mL (12.03 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 60.0 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: ≥ 6 mg/mL (12.03 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 60.0 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: ≥ 6 mg/mL (12.03 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 | 2.0055 mL | 10.0277 mL | 20.0554 mL | |
| 5 mM | 0.4011 mL | 2.0055 mL | 4.0111 mL | |
| 10 mM | 0.2006 mL | 1.0028 mL | 2.0055 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
