| Size | Price | |
|---|---|---|
| Other Sizes |
| Targets |
KRAS(G12C)
CFL-120 targets the KRAS G12C mutant protein, a frequently occurring oncogenic driver in various cancers, including non-small cell lung cancer (NSCLC). KRAS is a small GTPase that cycles between an inactive GDP-bound and an active GTP-bound state. The G12C mutation (glycine-to-cysteine substitution at codon 12) locks the protein in an active state, driving uncontrolled cell proliferation. CFL-120 is a covalent inhibitor that binds to the mutant cysteine residue (Cys12), trapping KRAS G12C in its inactive GDP-bound conformation and thereby blocking downstream signaling through the MAPK/ERK pathway. |
|---|---|
| ln Vitro |
H1792, SW1573, MiaPaca2, H358, A549, SW480, PANC-1, LCLC-103H, BxPC3, HCA-7, MRC-5, HUVEC-TERT, and CCD-exhibited antiproliferative effects after 72 hours of exposure to CFL-120. The IC50 values of 986Sk cells are 111.0, 23.6, 13.7, 16.9, 44.4, 14.8, 38.0, 30.0, 13.1, 9.8, 47.9, 24.4, 42.7 µM [1].
In vitro, CFL-120 demonstrates antiproliferative activity against a broad panel of cancer cell lines. In a 72-hour cell growth inhibition assay (MTT), it exhibited IC50 values of 11.0, 23.6, 13.7, 16.9, 44.4, 14.8, 38.0, 30.0, 13.1, 9.8, 47.9, 24.4, and 42.7 μM against H1792, SW1573, MiaPaca2, H358, A549, SW480, PANC-1, LCLC-103H, BxPC3, HCA-7, MRC-5, HUVEC-TERT, and CCD-986Sk cells, respectively. Notably, the compound shows activity against cells harboring different KRAS mutations, including G12C, G12D, and G12V. |
| ln Vivo |
CFL-120 (5 mg/kg for 5 treatments; 15 mg/kg for 5 treatments; 30 mg/kg for 3 treatments; ip) subcutaneous human lung carcinoma H1792 (KRasG12C mutant) and LCLC-103H (KRasWT)
In vivo, CFL-120 has demonstrated antitumor efficacy in mouse xenograft models. In a study using NOD/SCID female mice bearing subcutaneous human lung cancer tumors (H1792, KRasG12C mutant), intraperitoneal (i.p.) administration of CFL-120 at 5 mg/kg (5 treatments), 15 mg/kg (5 treatments), and 30 mg/kg (3 treatments) resulted in a 35.8% reduction in tumor growth compared to the control group. This confirms its potential as an in vivo-active compound for treating KRAS G12C-driven tumors. |
| Enzyme Assay |
The covalent binding and inhibitory activity of CFL-120 against KRAS G12C are assessed using biochemical assays. The compound's ability to covalently modify the target protein can be evaluated through mass spectrometry or gel-based assays. Its inhibitory potency is determined using in vitro kinase or GTPase activity assays, where recombinant KRAS G12C protein is incubated with varying concentrations of CFL-120. The inhibition of downstream effector binding (e.g., RAF) can be measured using TR-FRET (time-resolved fluorescence resonance energy transfer) or AlphaScreen technology. IC50 values are calculated from dose-response curves.
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| Cell Assay |
The cellular antiproliferative activity of CFL-120 is evaluated using a panel of cancer cell lines, particularly those with KRAS G12C mutations (e.g., H1792, SW1573, MIA PaCa-2, H358). Cells are seeded in 96-well plates and treated with serial dilutions of CFL-120 for 72 hours. Cell viability is then assessed using a standard colorimetric assay such as MTT or a luminescent assay like CellTiter-Glo. The half-maximal inhibitory concentration (IC50) is calculated by fitting the dose-response data to a sigmoidal curve. This assay confirms the compound's ability to inhibit the growth of KRAS-mutant cancer cells.
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| Animal Protocol |
Animal/Disease Models: NOD/SCID female mice (KRasWT (LCLC-103H) or KRasG12C (H1792) tumors) [1]
Doses: 5 mg/kg, 5 treatments; Growth of mouse tumor model [1]. 15 mg/kg, 5 treatments; 30 mg/kg, 3 treatments: intraperitoneal (ip) injection Experimental Results: Tumor growth was diminished by 35.8% compared to controls in the KRasG12C mutation model. In animal efficacy studies, CFL-120 is typically administered to immunodeficient mice bearing subcutaneous xenografts of human KRAS G12C-mutant cancer cell lines (e.g., H1792). Mice are randomized to receive either vehicle control or CFL-120 via intraperitoneal (i.p.) injection. Dosing schedules may vary; one study used 5 mg/kg for 5 treatments, 15 mg/kg for 5 treatments, and 30 mg/kg for 3 treatments. Tumor volume is measured twice weekly with calipers. At study termination, tumors are harvested for pharmacodynamic analysis, including assessment of target engagement and pathway modulation. |
| ADME/Pharmacokinetics |
The pharmacokinetic (PK) properties of CFL-120 have been characterized in NOD-SCID female mice following intraperitoneal administration at 15 mg/kg. Key parameters include: a maximum plasma concentration (Cmax) of 337 ± 123 ng/mL, time to reach Cmax (Tmax) of 0.25 hours, area under the curve (AUCt) of 169 ± 56 ng/mL*h, terminal half-life (t1/2) of 4.4 ± 0.6 hours, volume of distribution (Vd) of 12,096 ± 4000 mL, and clearance (CL) of 1895 ± 539 mL/h. These data indicate rapid absorption and a moderate half-life, supporting its use in in vivo efficacy studies.
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| Toxicity/Toxicokinetics |
Detailed toxicology studies for CFL-120 are limited in publicly available sources, as it is primarily a research tool. However, the compound is noted to have a favorable safety profile at efficacious doses in preclinical models. For instance, in the H1792 xenograft study, doses up to 30 mg/kg (administered 3 times) were tolerated, as evidenced by the ability to measure antitumor activity without reported overt toxicity. As a covalent inhibitor, its safety profile would need to be carefully evaluated in future development, considering potential off-target reactivity. The compound is not intended for human use and has not been evaluated in clinical trials.
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| References | |
| Additional Infomation |
CFL-120 (4,6-dichloroisatin) is a covalent KRasG12C inhibitor discovered through fragment-based drug discovery. It is a valuable chemical probe for studying the biology of KRAS G12C-mutant cancers and for validating the target in preclinical settings. Its low molecular weight and covalent mechanism of action provide a distinct pharmacological profile compared to other KRAS G12C inhibitors like ARS-1620. CFL-120 has potential for research in lung cancer and other KRAS G12C-driven malignancies. While it serves as a lead compound, its moderate potency (IC50 values in the micromolar range) suggests that further medicinal chemistry optimization is needed to develop a clinical candidate.
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| Molecular Formula |
C8H3CL2NO2
|
|---|---|
| Molecular Weight |
216.0209
|
| Exact Mass |
214.954
|
| CAS # |
18711-15-4
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| Related CAS # |
18711-15-4
|
| PubChem CID |
251908
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| Appearance |
Light yellow to yellow solid
|
| Density |
1.6±0.1 g/cm3
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| Melting Point |
260.5-261°C
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| Index of Refraction |
1.638
|
| LogP |
2.2
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| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
0
|
| Heavy Atom Count |
13
|
| Complexity |
269
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
ClC1=C([H])C(=C([H])C2=C1C(C(N2[H])=O)=O)Cl
|
| InChi Key |
CGCVHJCZBIYRQC-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C8H3Cl2NO2/c9-3-1-4(10)6-5(2-3)11-8(13)7(6)12/h1-2H,(H,11,12,13)
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| Chemical Name |
4,6-dichloro-1H-indole-2,3-dione
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| Synonyms |
CFL-120
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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 |
| 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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.6292 mL | 23.1460 mL | 46.2920 mL | |
| 5 mM | 0.9258 mL | 4.6292 mL | 9.2584 mL | |
| 10 mM | 0.4629 mL | 2.3146 mL | 4.6292 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.
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