| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 500mg | |||
| 1g | |||
| Other Sizes |
| Targets |
KRAS(G12C)
ARS-1323-alkyne targets the switch-II pocket (S-IIP) of the KRAS G12C mutant protein. The switch-II pocket is a shallow, hydrophobic groove on the surface of KRAS that is only exposed when the protein is in the inactive GDP-bound conformation. This pocket has been exploited for the development of covalent KRAS G12C inhibitors such as sotorasib and adagrasib, which bind to the mutant cysteine residue (Cys12) within this pocket. ARS-1323-alkyne covalently binds to Cys12 of KRAS G12C, trapping the protein in the inactive GDP-bound state. The alkyne functionality of the compound allows for the conjugation of various tags (fluorescent probes, biotin, affinity handles) via click chemistry, enabling the detection and visualization of KRAS G12C occupancy and activity in cells. |
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| ln Vitro |
In vitro, ARS-1323-alkyne demonstrates covalent binding to KRAS G12C with high specificity. The compound selectively modifies the mutant cysteine residue (Cys12) in KRAS G12C, with minimal labeling of wild-type KRAS or other cysteine-containing proteins. The compound's ability to serve as a chemical reporter is demonstrated by its use in competitive labeling experiments, where the binding of ARS-1323-alkyne to KRAS G12C is reduced in the presence of other KRAS G12C inhibitors, providing a measure of target occupancy. The compound does not exhibit significant cytotoxicity in cell-based assays at concentrations used for labeling experiments (typically 0.1-10 μM). The clickable alkyne group enables the conjugation of fluorophores or affinity tags for detection by fluorescence microscopy, flow cytometry, or mass spectrometry.
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| ln Vivo |
In vivo, ARS-1323-alkyne is used as a chemical probe to study KRAS G12C occupancy and activity in tumor tissues. Following administration to mice bearing KRAS G12C-mutant xenografts, the compound covalently labels KRAS G12C in tumor tissue, allowing for the assessment of target engagement. The labeled protein can be detected by conjugation with azide-functionalized probes ex vivo, enabling quantification of KRAS G12C occupancy. This approach has been used to validate the target engagement of KRAS G12C inhibitors in preclinical models and to study the pharmacodynamics of KRAS G12C inhibition. The compound's utility in vivo is limited by its role as a probe rather than a therapeutic agent, and it is primarily used for mechanistic studies rather than efficacy evaluation.
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| Enzyme Assay |
The binding of ARS-1323-alkyne to KRAS G12C is assessed using biochemical assays. Recombinant KRAS G12C protein (GDP-bound) is incubated with varying concentrations of ARS-1323-alkyne (0.01-10 μM) in assay buffer. The covalent labeling of KRAS G12C is detected by SDS-PAGE followed by in-gel fluorescence after conjugation with an azide-fluorophore via click chemistry. The extent of labeling is quantified by densitometry or fluorescence scanning. The selectivity of labeling is assessed by testing the compound against wild-type KRAS, other RAS isoforms (HRAS, NRAS), and a panel of cysteine-containing proteins. The compound's ability to compete with other KRAS G12C inhibitors (e.g., sotorasib, adagrasib) for binding to KRAS G12C is assessed in competitive labeling assays.
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| Cell Assay |
Cellular activity of ARS-1323-alkyne is evaluated in KRAS G12C-mutant cancer cell lines such as NCI-H358 and MIA PaCa-2. Cells are treated with ARS-1323-alkyne at concentrations ranging from 0.1 to 10 μM for 1-24 hours. After treatment, cells are lysed, and the lysates are subjected to click chemistry conjugation with azide-functionalized fluorescent probes or biotin. Labeled KRAS G12C is detected by in-gel fluorescence, Western blot (with streptavidin-HRP for biotin conjugates), or mass spectrometry. The compound is used in competitive labeling assays to assess the target occupancy of other KRAS G12C inhibitors, where cells are pre-treated with the inhibitor of interest followed by ARS-1323-alkyne labeling. The reduction in labeling indicates the degree of target occupancy by the inhibitor. Cell viability is assessed to ensure that labeling experiments are performed at non-cytotoxic concentrations.
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| Animal Protocol |
In animal studies, ARS-1323-alkyne is administered to immunodeficient mice bearing subcutaneous xenografts of KRAS G12C-mutant NSCLC cell lines. Mice are treated with ARS-1323-alkyne at doses of 10-50 mg/kg via intraperitoneal or intravenous injection. After 1-6 hours, tumors are harvested and lysed. The lysates are subjected to click chemistry conjugation with azide-functionalized fluorescent probes, and labeled KRAS G12C is detected by in-gel fluorescence or Western blot. The compound is used in competitive labeling experiments in vivo, where mice are pre-treated with a KRAS G12C inhibitor of interest followed by ARS-1323-alkyne to assess target occupancy in tumor tissue. This approach provides valuable pharmacodynamic data for the development of KRAS G12C inhibitors.
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| ADME/Pharmacokinetics |
Pharmacokinetic studies of ARS-1323-alkyne are limited as the compound is a research probe rather than a therapeutic candidate. Following intraperitoneal or intravenous administration in rodents at 10-50 mg/kg, the compound achieves sufficient exposure in tumor tissue to enable covalent labeling of KRAS G12C. The compound is rapidly cleared from circulation with a half-life of 1-3 hours, consistent with its use as a short-acting labeling reagent. The compound's distribution to tissues including tumor tissue is sufficient for labeling applications. As a research probe, comprehensive PK studies are not typically performed, and the compound is used primarily for mechanistic studies.
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| Toxicity/Toxicokinetics |
Toxicology studies of ARS-1323-alkyne are limited as the compound is a research tool rather than a therapeutic agent. In short-term studies at doses used for labeling experiments (10-50 mg/kg), the compound is generally well-tolerated with no significant adverse effects on body weight, food consumption, or general health. The compound is not intended for human use and has not been evaluated in clinical trials. Researchers should follow standard safety precautions when handling the compound, including the use of appropriate personal protective equipment and adherence to institutional safety guidelines.
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| References | |
| Additional Infomation |
ARS-1323-alkyne is a versatile chemical tool for studying KRAS G12C biology and for validating the target engagement of KRAS G12C inhibitors. The compound's combination of covalent binding to KRAS G12C and clickable alkyne functionality makes it a valuable probe for a range of applications, including: (1) assessing target occupancy of KRAS G12C inhibitors in cells and tissues; (2) visualizing the localization and dynamics of KRAS G12C in living cells; (3) identifying KRAS G12C-interacting proteins and downstream signaling components; and (4) studying the mechanisms of resistance to KRAS G12C inhibitors. ARS-1323-alkyne is widely used in KRAS drug discovery programs and has contributed to the development of clinically approved KRAS G12C inhibitors. The compound is available from commercial suppliers for research purposes and is cited in numerous peer-reviewed publications in the RAS field.
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| Molecular Formula |
C28H27CLF2N6O3
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|---|---|
| Exact Mass |
568.18
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| Elemental Analysis |
C, 59.10; H, 4.78; Cl, 6.23; F, 6.68; N, 14.77; O, 8.44
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| CAS # |
2436544-27-1
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| Related CAS # |
ARS-1323;1698024-73-5
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| PubChem CID |
138454781
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| Appearance |
Off-white to light brown solid powder
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| LogP |
4.4
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
40
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| Complexity |
960
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN(CC#C)C(=O)CCNC1=NC2=C(C(=C(C=C2C(=N1)N3CCN(CC3)C(=O)C=C)Cl)C4=C(C=CC=C4F)O)F
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| InChi Key |
FCCFZQSNDSLLMI-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C28H27ClF2N6O3/c1-4-11-35(3)22(40)9-10-32-28-33-26-17(27(34-28)37-14-12-36(13-15-37)21(39)5-2)16-18(29)23(25(26)31)24-19(30)7-6-8-20(24)38/h1,5-8,16,38H,2,9-15H2,3H3,(H,32,33,34)
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| Chemical Name |
3-[[6-chloro-8-fluoro-7-(2-fluoro-6-hydroxyphenyl)-4-(4-prop-2-enoylpiperazin-1-yl)quinazolin-2-yl]amino]-N-methyl-N-prop-2-ynylpropanamide
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| Synonyms |
ARS1323-Alkyne; ARS-1323 Alkyne; ARS 1323-Alkyne; ARS-1323-Alkyne
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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) |
DMSO: ~250 mg/mL (~439.4 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (3.66 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 (3.66 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
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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