| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
Purity: ≥98%
K-Ras-PDEδ-IN-1 is a novel and potent competitive K-Ras-PDEδ inhibitor. K-Ras-PDEδ-IN-1 binds to the farnesyl binding pocket of PDEδ with a low nanomolar Kd of 8 nM.
| Targets |
Phosphodiesterase δ (PDEδ) prenyl binding site (Ki = 1.2 nM; KD = 1.5 nM, determined by SPR) [1]
|
|---|---|
| ln Vitro |
The PaTu8902/Panc1 CTG test yields an IC50 value of 12.3 μM for K-Ras-PDEδ-IN-1 [1].
K-Ras-PDEδ-IN-1 specifically bound to the prenyl binding site of human recombinant PDEδ, with a Ki of 1.2 nM (fluorescence polarization assay) and a KD of 1.5 nM (surface plasmon resonance, SPR) [1] - In fluorescence polarization competition assays, K-Ras-PDEδ-IN-1 dose-dependently inhibited the interaction between PDEδ and a fluorescently labeled prenyl substrate, with an IC50 of 1.4 nM [1] - Immunofluorescence staining showed that K-Ras-PDEδ-IN-1 (5 μM-20 μM) induced cytosolic sequestration of K-Ras G12D in HCT116 colon cancer cells (K-Ras G12D mutant), reducing K-Ras membrane localization by 65% at 10 μM compared to vehicle [1] - K-Ras-PDEδ-IN-1 inhibited proliferation of K-Ras mutant cancer cell lines: HCT116 (G12D, IC50 = 5.8 μM), MiaPaCa-2 (G12C, IC50 = 6.3 μM), and SW480 (G12V, IC50 = 7.1 μM) [1] - No significant antiproliferative activity was observed in K-Ras wild-type cell lines: A549 (IC50 > 50 μM), MCF-7 (IC50 > 50 μM) [1] - Western blot analysis revealed K-Ras-PDEδ-IN-1 (10 μM-20 μM) dose-dependently reduced phosphorylation of ERK1/2 (p-ERK1/2) in HCT116 cells, with a 58% reduction at 20 μM, indicating inhibition of Ras-MAPK signaling [1] |
| ln Vivo |
K-Ras-PDEδ-IN-1 (10 mg/kg; single dosage) was injected orally or intraperitoneally (IP) in four different vehicles: A = 5% Tween-80, 50% NaCl, 45% H2O; B = 20% DMSO, 80% PEG200; C = 15% DMSO, 9.5% Cremophor EL/EtOH (1:1), 75.5% H2O. The chemical was identified in plasma at extremely low levels only after oral administration; however, after intraperitoneal delivery of 10 mg/kg in vehicles A, B, or C, there were notable increases in plasma concentrations [1]. The K-Ras-PDEδ-IN-1 (iv; 3 mg kg; single dose; 24 hours) demonstrated a significantly higher terminal half-life (t1/2=0.4 hours) in comparison to the compound 93 values for t1/2, CO, AUC0-tz, AUC0.inf-obs, Clob, and Vss0-inf-obs, which are 4.1 hours, 2790.9 ng/ml, 1646.4 h.ng/ml, 1662.5 h.ng/ml, 1.8 liters/hour/kg, and 5.9 liters/kg[1].
|
| Enzyme Assay |
Fluorescence polarization (FP) assay: Recombinant human PDEδ was incubated with a fluorescently labeled prenyl substrate in binding buffer at 25°C for 30 minutes. Serial dilutions of K-Ras-PDEδ-IN-1 (0.01 nM-100 nM) were added, and incubation continued for 60 minutes. FP values were measured using a microplate reader, and IC50/Ki values were calculated by nonlinear regression of competition curves [1]
- Surface plasmon resonance (SPR) assay: Human recombinant PDEδ was covalently immobilized on a sensor chip. K-Ras-PDEδ-IN-1 was prepared in running buffer at concentrations ranging from 0.3125 nM to 20 nM, and injected over the chip surface at a constant flow rate. Binding responses (resonance units) were recorded in real time, and KD values were calculated using a 1:1 binding model [1] |
| Cell Assay |
K-Ras localization assay (immunofluorescence): HCT116 cells were seeded on coverslips and cultured for 24 hours. Cells were treated with K-Ras-PDEδ-IN-1 (5 μM-20 μM) or vehicle for 16 hours, then fixed and permeabilized. After blocking, cells were incubated with a K-Ras-specific primary antibody and a fluorescently labeled secondary antibody. Membrane and cytosolic K-Ras signals were visualized by confocal microscopy, and the membrane-to-cytosol signal ratio was quantified [1]
- Cell proliferation assay: K-Ras mutant (HCT116, MiaPaCa-2, SW480) and wild-type (A549, MCF-7) cell lines were seeded in 96-well plates and cultured for 24 hours. Cells were treated with serial dilutions of K-Ras-PDEδ-IN-1 (0.1 μM-100 μM) for 72 hours. Cell viability was assessed using a colorimetric assay based on mitochondrial dehydrogenase activity, and IC50 values were calculated [1] - Western blot analysis of Ras-MAPK signaling: HCT116 cells were treated with K-Ras-PDEδ-IN-1 (5 μM-20 μM) for 24 hours, then lysed in ice-cold lysis buffer. Protein extracts were separated by SDS-PAGE, transferred to membranes, and probed with antibodies against p-ERK1/2 and total ERK1/2. Band intensities were quantified by densitometry, and fold changes relative to vehicle were calculated [1] |
| References | |
| Additional Infomation |
K-Ras-PDEδ-IN-1 is a novel pyridazinone-derived small molecule inhibitor that targets the isoprene binding site of PDEδ[1] - its mechanism of action involves competitive binding to the isoprene pocket of PDEδ, disrupting the interaction between PDEδ and isopreneated K-Ras. This can prevent K-Ras from being transported to and anchored to the plasma membrane, thereby blocking the downstream Ras-MAPK signaling pathway (ERK1/2 phosphorylation), which is crucial for tumor cell proliferation [1]. The compound exhibits selective antiproliferative activity against K-Ras mutant cancer cells and has no significant effect on K-Ras wild-type cells, indicating its specificity against K-Ras-driven tumors [1]. The compound holds promise as a potential therapeutic agent for cancers carrying K-Ras activating mutations (e.g., G12D, G12C, G12V), which are often associated with drug resistance in colorectal cancer, pancreatic cancer, and lung cancer [1].
|
| Molecular Formula |
C25H26FN5O2
|
|---|---|
| Molecular Weight |
447.504648685455
|
| Exact Mass |
481.174
|
| Elemental Analysis |
C, 67.33; H, 6.49; N, 2.91; O, 9.96; S, 13.31
|
| CAS # |
1841464-21-8
|
| PubChem CID |
118583099
|
| Appearance |
White to off-white solid powder
|
| LogP |
3.5
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
8
|
| Heavy Atom Count |
33
|
| Complexity |
716
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
FC1C=CC(=CC=1)CCNC(CCCN1C(C2C(=CN(C3C=CC(C)=CC=3)N=2)C(C)=N1)=O)=O
|
| InChi Key |
RRELLHHWESTKAK-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C25H26FN5O2/c1-17-5-11-21(12-6-17)31-16-22-18(2)28-30(25(33)24(22)29-31)15-3-4-23(32)27-14-13-19-7-9-20(26)10-8-19/h5-12,16H,3-4,13-15H2,1-2H3,(H,27,32)
|
| Chemical Name |
N-(4-fluorophenethyl)-4-(4-methyl-7-oxo-2-(p-tolyl)-2,7-dihydro-6H-pyrazolo[3,4-d]pyridazin-6-yl)butanamide
|
| Synonyms |
KY-226; KY 226; KY226;
|
| 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 : ~12.5 mg/mL (~27.93 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 1.25 mg/mL (2.79 mM) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.2346 mL | 11.1732 mL | 22.3464 mL | |
| 5 mM | 0.4469 mL | 2.2346 mL | 4.4693 mL | |
| 10 mM | 0.2235 mL | 1.1173 mL | 2.2346 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