KL-50
Based on 1 Customer Validation
KL-50 is an orally active N3-(2-fluoroethyl) imidazotetrazine DNA alkylating agent with selectivity for MGMT-deficient cells. KL-50 introduces a 2-fluoroethyl group at the O6 position of guanine, forming O6-(2-fluoroethyl) guanine (O6FEtG) lesions and further generating DNA interstrand crosslinks (ICLs). KL-50 induces DNA double-strand breaks, activates ATR-CHK1 and ATM-CHK2 DNA damage responses, and causes cell cycle arrest and micronucleus formation. KL-50 can be used for glioblastoma and DNA damage response research.
Nur für Forschungszwecke. Wir verkaufen nicht an Patienten.
- Reinheit : 99.82%
- CAS. Nr.: 1161826-19-2
- Formel: C7H7FN6O2
- Molecular Weight:226.17
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Speicherung:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
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Biologische Aktivität
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Chemical Information
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Lösungsmittel & Löslichkeit
- Protokoll
- Reinheit & Dokumentation
- Verweise
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Help & FAQs
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Biologische Aktivität
Beschreibung
In Vitro
KL-50 (4a) (200 μM; 2-24 h) produces time-dependent DNA interstrand crosslinks in MGMT−/MMR− LN229 cells, with DNA migration in the IR-modified comet assay significantly decreasing from 8 h and showing the greatest change at 8-24 h[1].
KL-50 (20 μM; 2-48 h) activates CHK1/CHK2-related DNA damage response in MGMT-deficient LN229 cells and induces γH2AX, 53BP1, and pRPA DNA damage foci; this response is independent of MMR status[1].
KL-50 (10-500 μM; 5 days) produces cytotoxic activity in post-TMZ, MMR-deficient GBM6R-m185 cells with a cell-death IC50 of 9.4 μM[2].
KL-50 (10-500 μM; 5 days) reduces the number of viable GBM6R-m185 cells, with a live-cell-count IC50 of 6.6 μM[2].
KL-50 (10-500 μM; 5 days) exhibits cell-death IC50 values of 27.2 μM (MMR+) and 13 μM (MMR−) in GBM6 cells, and 53.1 μM (MMR+) and 9.4 μM (MMR−) in GBM12 cells, indicating that MSH6 loss does not confer KL-50 tolerance[2].
KL-50 (10-500 μM; 5 days) exhibits live-cell-count IC50 values of 7.4 μM (MMR+) and 6.5 μM (MMR−) in GBM6 cells, and 9.7 μM (MMR+) and 8.5 μM (MMR−) in GBM12 cells[2].
KL-50 (compound 1) (30 or 200 μM) selectively inhibits the long-term survival of MGMT-deficient cells in clonogenic assays using isogenic LN229 GBM cells; at 30 μM, the surviving fractions of MGMT−/MMR+ and MGMT−/MMR− cells are 6.3 × 10−4 and 4.0 × 10−5, respectively, whereas MGMT+ cells show only approximately 60-70% cell kill at 200 μM[3].
KL-50 (91 nM-200 μM; 7 doses) exhibits MGMT expression-dependent differences in cell viability in the PRISM screen of 902 human cancer cell lines, with MGMT expression positively correlating with cell viability, and MGMT expression is the most prominent determinant of sensitivity in this screen[3].
KL-50 (200 μM; 24 h) reduces the %DNA in tail in the modified alkaline comet assay only in MGMT−/MMR± LN229 cells, supporting that KL-50 forms DNA ICLs in the MGMT-deficient background[3].
KL-50 (1 mM; 5 or 72 h) forms O6FEtdG in calf thymus DNA at both 5 h and 72 h, whereas [dG (N1)-dC (N3)]Et ICL is detected only at 72 h, supporting a mechanism in which O6FEtG forms first and is slowly converted to DNA ICL[3].
KL-50 (200, 500 μM; 8 h) does not form detectable DNA-MGMT cross-links in MGMT+ DLD1 cells, consistent with its lower cross-linking damage in the MGMT-positive background[3].
KL-50 (20 μM; 2-48 h) progressively induces G2 phase arrest from 24-48 h in MGMT−/MMR+ LN229 cells, whereas G2 phase arrest is attenuated in MGMT−/MMR− cells[1].
KL-50 (50 μM; 5 days) induces DNA double-strand break-related p-H2AX and p-p95 signals in both MMR-proficient and MSH6-KO GBM6 and GBM12 cells[2].
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:LN229 MGMT−/MMR±
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Concentration:20 μM
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Incubation Time:2, 8, 24, 48 h
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Result:Progressively increased G2 arrest from 24 to 48 h in MGMT−/MMR+ cells.
Produced attenuated G2 arrest in MGMT−/MMR− cells.
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Cell Line:GBM6R-m185
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Concentration:10, 50, 100, 250, 500 μM
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Incubation Time:5 days
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Result:Induced cell death with an IC50 of 9.4 μM.
Was approximately 3-fold more potent than TMZ by the cell-death endpoint.
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Cell Line:GBM6, GBM12
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Concentration:50 μM
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Incubation Time:5 days
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Result:Increased the DNA DSB-associated signals p-H2AX and p-p95 in both MMR-proficient and MMR-deficient cells.
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Cell Line:LN229 MGMT+/−, MMR+/− cells
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Concentration:20 μM
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Incubation Time:48 h
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Result:Treatment caused cell cycle arrest, with a significant increase in G2 phase cells.
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Cell Line:DLD1
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Concentration:200, 500 μM
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Incubation Time:8 h
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Result:Did not produce detectable DNA-MGMT cross-links.
In Vivo
KL-50 (10 mg/kg; oral gavage; 3 consecutive weeks; starting on day 11 post-engraftment) extends the median survival of intracranial GBM6 PDX NSG mice to 57 days, compared with 32.5 and 34 days for the vehicle and 4 Gy RT groups, respectively[2].
KL-50 (10 mg/kg; oral gavage; 3 consecutive weeks; starting on day 11 post-engraftment) extends the median survival of intracranial GBM12 PDX NSG mice to 71 days, compared with 33 and 36.5 days for the vehicle and 4 Gy RT groups, respectively; in combination with 4 Gy fractionated RT, the median survival further reaches 80 days[2].
KL-50 (10 mg/kg; oral gavage; 3 consecutive weeks; starting on day 11 post-engraftment) extends the median survival of post-TMZ, MMR-deficient GBM6R-m185 intracranial PDX NSG mice to 140 days, compared with 37 and 108 days for the vehicle and TMZ groups, respectively[2].
KL-50 (25-200 mg/kg; single dose) causes >10% body weight loss at 100 and 200 mg/kg in healthy mice, recovering to baseline after approximately 1 week; 2 of 3 mice in the 200 mg/kg group show marked intolerance, whereas blood cell counts reveal no significant toxic changes[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:LN229 MGMT−/MMR+ and LN229 MGMT−/MSH6− flank tumors; starting tumor volume approximately 350-400 mm3[1]
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Dosage:25 mg/kg
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Administration:p.o.; Monday, Wednesday, and Friday; 3 weeks
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Result:Persistently suppressed growth of both MGMT−/MMR+ and MGMT−/MSH6− flank tumors.
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Animal Model:Intracranial LN229 MGMT−/MMR− xenograft[1]
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Dosage:25 mg/kg
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Administration:i.p.; Monday-Friday; 1 week
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Result:Suppressed intracranial tumor growth.
Produced more sustained tumor control than the TMZ comparator.
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Animal Model:8-10-week-old NSG mice; equal male and female numbers; intracranial implantation of 100,000 GBM6 PDX cells into the right frontal lobe[2]
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Dosage:10 mg/kg
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Administration:p.o.; Monday-Friday; 3 consecutive weeks; starting day 11 post-engraftment
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Result:Extended median survival to 57 days versus 32.5 days for vehicle and 34 days for RT.
Combined use with 4 Gy RT produced a median survival of 63 days.
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Animal Model:8-10-week-old NSG mice; equal male and female numbers; intracranial implantation of 100,000 GBM12 PDX cells into the right frontal lobe[2]
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Dosage:10 mg/kg
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Administration:p.o.; Monday-Friday; 3 consecutive weeks; starting day 11 post-engraftment
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Result:Extended median survival to 71 days versus 33 days for vehicle and 36.5 days for RT.
Combined use with 4 Gy RT extended median survival to 80 days.
Chemical Information
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CAS. Nr. 1161826-19-2
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Appearance Solid
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Molecular Weight 226.17
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Formel C7H7FN6O2
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Color Off-white to pink
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SMILES
O=C(C(N=CN12)=C2N=NN(CCF)C1=O)N
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Versand
Room temperature in continental US; may vary elsewhere.
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Speicherung
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Lösungsmittel & Löslichkeit
In Vitro:
DMSO : 100 mg/mL (442.15 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Konzentration (Stammlösung) × Volumen (Stammlösung) = Konzentration (Ziellösung) × Volumen (Ziellösung)
Protokoll
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Flow cytometric DNA-content cell-cycle staining
Flow cytometric DNA-content cell-cycle staining measures the fluorescence intensity of DNA-bound fluorochromes in single cells or nuclei to estimate DNA content distributions, allowing assignment of populations to G0/G1, S, and G2/M phases by DNA histogram deconvolution. Propidium iodide (PI) intercalates into DNA, and PI fluorescence is proportional to cellular DNA content when staining is performed under conditions that make DNA accessible and minimize non-DNA signal. Cells with G2/M DNA content are expected to show approximately twice the fluorescence intensity of G0/G1 cells, while S-phase cells occupy intermediate fluorescence values. PI-based DNA-content analysis can also detect cells with fractional DNA content, often reported as sub-G1, when DNA fragmentation and extraction during staining reduce retained DNA signal in apoptotic cells. DAPI is an alternative DNA fluorochrome for univariate DNA-content analysis, while bivariate approaches combining DNA content with proliferation
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BrdU Incorporation Assay
Bromodeoxyuridine (BrdU) incorporation assay is based on the principle that BrdU, a thymidine analog, is incorporated into newly synthesized DNA during the S phase of the cell cycle, thereby serving as a marker of DNA replication and cellular proliferation. Incorporated BrdU can be detected using anti-BrdU antibodies following DNA denaturation, enabling visualization or quantification of proliferating cells through immunochemical detection methods such as immunofluorescence or immunohistochemistry.
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Protocol for Cell Cycle
Cell-cycle analysis by flow cytometry measures DNA content in single cells to estimate the fraction of cells in G0/G1, S, and G2/M phases. Propidium iodide intercalates into DNA, and after RNA removal with RNase, fluorescence intensity reflects cellular DNA content: 2N cells are assigned to G0/G1, cells between 2N and 4N to S phase, and 4N cells to G2/M. DNA-content analysis alone cannot reliably separate G0 from G1 or G2 from M. Ki-67 can distinguish quiescent G0 cells from cycling cells, EdU or BrdU incorporation marks active DNA synthesis in S phase, and phospho-histone H3 staining identifies mitotic cells within the 4N population.
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Genotoxicity/Mutagenicity Study
The bacterial reverse mutation assay detects point mutations that restore amino-acid prototrophy in auxotrophic Salmonella typhimurium or Escherichia coli tester strains; after exposure to a test article, mutagenic activity is read out as an increased number of revertant colonies on minimal agar compared with the vehicle control. The assay uses tester strains with different mutation targets so that base-substitution and frameshift mutagens can be detected, and testing is performed with and without exogenous mammalian metabolic activation because some chemicals require biotransformation to become mutagenic.
Reinheit & Dokumentation
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Data Sheet (294 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
Verweise
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 4.4215 mL | 22.1073 mL | 44.2145 mL | 110.5363 mL |
| 5 mM | 0.8843 mL | 4.4215 mL | 8.8429 mL | 22.1073 mL | |
| 10 mM | 0.4421 mL | 2.2107 mL | 4.4215 mL | 11.0536 mL | |
| 15 mM | 0.2948 mL | 1.4738 mL | 2.9476 mL | 7.3691 mL | |
| 20 mM | 0.2211 mL | 1.1054 mL | 2.2107 mL | 5.5268 mL | |
| 25 mM | 0.1769 mL | 0.8843 mL | 1.7686 mL | 4.4215 mL | |
| 30 mM | 0.1474 mL | 0.7369 mL | 1.4738 mL | 3.6845 mL | |
| 40 mM | 0.1105 mL | 0.5527 mL | 1.1054 mL | 2.7634 mL | |
| 50 mM | 0.0884 mL | 0.4421 mL | 0.8843 mL | 2.2107 mL | |
| 60 mM | 0.0737 mL | 0.3685 mL | 0.7369 mL | 1.8423 mL | |
| 80 mM | 0.0553 mL | 0.2763 mL | 0.5527 mL | 1.3817 mL | |
| 100 mM | 0.0442 mL | 0.2211 mL | 0.4421 mL | 1.1054 mL |
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