RSL3-NH2
RSL3-NH2 is a GPX4 inhibitor and ferroptosis inducer. RSL3-NH2 triggers the iron-dependent cell death pathway associated with lipid peroxidation by inhibiting GPX4 activity. RSL3-NH2 exhibits significant cytotoxicity against colorectal cancer cells and effectively induces their ferroptosis. RSL3-NH2 can serve as a ADC payload for synthesizing antibody-drug conjugates (ADC) and be used in colorectal cancer-related research.
For research use only. We do not sell to patients.
- CAS No.: 3051815-39-2
- Formula: C21H20ClN3O3
- Molecular Weight:397.85
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocols
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Description
IC50 & Target
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GPX4 |
In Vitro
The ADC formed by conjugating RSL3-NH2 with a bispecific antibody, namely CDH17 x GUCY2C-RSL3-NH2 ADC, potently inhibits the proliferation of SW1463 and LS1034 cells and induces significant MDA and lipid ROS accumulation, with IC50 values of 4.97 μg/mL and 2.249 μg/mL at 72 h, respectively[1].
CDH17 x GUCY2C-RSL3-NH2 ADC (5 μg/mL; 72 h) exhibits bystander cytotoxicity against CDH17/GUCY2C-negative HT-29 cells when co-cultured with CDH17/GUCY2C-positive SW1463 or LS1034 cells[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:SW1463/HT-29 co-cultures, LS1034/HT-29 co-cultures
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Concentration:5 μg/mL
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Incubation Time:72 h
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Result:Increased bystander killing of CDH17/GUCY2C-negative HT-29 cells in co-culture with positive cells, compared to the control group.
In Vivo
CDH17 x GUCY2C-RSL3-NH2 ADC (1 mg/kg; intravenous injection; once every 5 days; 7 injections in total) induces significant regression of LS1034 colorectal tumors in nude mice and triggers strong ferroptosis (elevated levels of MDA and 4-HNE)[1].
CDH17 x GUCY2C-RSL3-NH2 ADC (1 mg/kg; intravenous injection; once every 5 days; 7 injections total) exhibits favorable safety in double-humanized BALB/c mice, with no hepatotoxicity or organ damage detected[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:hCDH17/hGUCY2 double-humanized BALB/c (7-week-old female)[1]
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Dosage:1 mg/kg
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Administration:i.v.; once every 5 days; 7 injections total
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Result:Maintained serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) within the normal range, not significantly different from PBS control.
Showed no histopathological signs of toxicity in the heart, liver, spleen, lungs, or kidneys.
Chemical Information
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CAS No. 3051815-39-2
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Molecular Weight 397.85
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Formula C21H20ClN3O3
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SMILES
NC(C=C1)=CC=C1[C@H](N2C(CCl)=O)C(NC3=C4C=CC=C3)=C4C[C@@H]2C(OC)=O
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Please store the product under the recommended conditions in the Certificate of Analysis.
Solvent & Solubility
In Vitro:
Ethanol : 100 mg/mL (251.35 mM; Need ultrasonic)
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% EtOH 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (6.28 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% EtOH 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (6.28 mM); Suspended solution
This protocol yields a suspended solution of ≥ 2.5 mg/mL (saturation unknown). Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Add each solvent one by one: 10% EtOH 90% Corn Oil
Solubility: ≥ 2.5 mg/mL (6.28 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 900 μL Corn oil, and mix evenly.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Working solution concentration: 0.22 mg/mL
Protocols
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Mammalian live/dead viability and cytotoxicity staining
Live/dead viability and cytotoxicity staining assays are based on the simultaneous detection of intracellular esterase activity in metabolically active (viable) cells and membrane integrity loss in non-viable cells. In commonly used dual-staining approaches, membrane-permeant fluorogenic substrates are converted by intracellular esterases into fluorescent products in live cells, while impermeant DNA-binding dyes selectively enter cells with compromised plasma membranes and label nucleic acids in dead or dying cells, enabling discrimination between viable and non-viable populations by fluorescence microscopy or flow cytometry.
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Apoptosis Solutions
Apoptosis is a regulated, generally non-lytic cell-death pathway that removes unwanted, damaged, infected, or abnormal cells through coordinated morphological changes, caspase activation, DNA fragmentation, and membrane remodeling. The intrinsic apoptosis pathway is controlled mainly by mitochondrial outer membrane permeabilization, BCL-2 family proteins, cytochrome c release, apoptosome formation, caspase-9 activation, and downstream executioner caspase-3/7 activation. The extrinsic apoptosis pathway is initiated by death receptors such as Fas, TNFR, and TRAIL receptors, which recruit adaptor proteins and activate caspase-8 before engaging executioner caspases or mitochondrial amplification through BID cleavage. Apoptosis is linked to many phenotypes, including cancer cell killing, tissue homeostasis, immune regulation, neurodegeneration, infection response, and treatment-induced cytotoxicity; unresolved questions include how apoptosis interacts with necroptosis, pyroptosis, ferroptos
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Ferroptosis Solutions
Ferroptosis is an iron-dependent, non-apoptotic form of regulated cell death characterized by lethal lipid peroxidation and sensitivity to suppression by iron chelators or lipophilic radical-trapping antioxidants. The core pathway links cystine uptake through system Xc−, glutathione availability, GPX4-dependent detoxification of phospholipid hydroperoxides, iron-dependent oxidative reactions, and polyunsaturated-phospholipid metabolism into a cell-death program that is biochemically and morphologically distinct from apoptosis, necrosis, and autophagy. The ferroptosis pathway is experimentally linked to phenotype through chemical and genetic perturbation. Erastin induces ferroptosis by inhibiting cystine uptake through system Xc− and weakening antioxidant defenses, while GPX4 inhibition or depletion causes lipid peroxide accumulation and ferroptotic cancer-cell death. ACSL4 and oxidizable arachidonoyl- or adrenoyl-containing phosphatidylethanolamines shape ferroptosis sensitivity by con
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Cell Viability Determination by MTT Colorimetric Assay
The following protocol uses the MTT colorimetric assay as a classic literature-established method for assessing cell viability/metabolic activity in cultured mammalian cells. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by metabolically active cells to a colored formazan product; the amount of formazan is quantified spectrophotometrically and provides an indirect measure of metabolically active viable cells. Importantly, MTT reduction reflects cellular oxidoreductase/metabolic activity rather than an absolute direct count of living cells, so changes in cellular metabolism can alter the signal independently of cell number.
Purity & Documentation
References
Complete Stock Solution Preparation Table
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
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| Ethanol | 1 mM | 2.5135 mL | 12.5676 mL | 25.1351 mL | 62.8378 mL |
| 5 mM | 0.5027 mL | 2.5135 mL | 5.0270 mL | 12.5676 mL | |
| 10 mM | 0.2514 mL | 1.2568 mL | 2.5135 mL | 6.2838 mL | |
| 15 mM | 0.1676 mL | 0.8378 mL | 1.6757 mL | 4.1892 mL | |
| 20 mM | 0.1257 mL | 0.6284 mL | 1.2568 mL | 3.1419 mL | |
| 25 mM | 0.1005 mL | 0.5027 mL | 1.0054 mL | 2.5135 mL | |
| 30 mM | 0.0838 mL | 0.4189 mL | 0.8378 mL | 2.0946 mL | |
| 40 mM | 0.0628 mL | 0.3142 mL | 0.6284 mL | 1.5709 mL | |
| 50 mM | 0.0503 mL | 0.2514 mL | 0.5027 mL | 1.2568 mL | |
| 60 mM | 0.0419 mL | 0.2095 mL | 0.4189 mL | 1.0473 mL | |
| 80 mM | 0.0314 mL | 0.1571 mL | 0.3142 mL | 0.7855 mL | |
| 100 mM | 0.0251 mL | 0.1257 mL | 0.2514 mL | 0.6284 mL |
RSL3-NH2 Related Classifications
HY-175615 Related Classifications
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