FM213
FM213 is a PD-L1 inhibitor with a human IC50 of 323 nM. FM213 blocks PD-1/PD-L1 protein-protein interactions, and promotes endocytosis and lysosome-dependent degradation of PD-L1 on the surface of cancer cells. FM213 binds to PD-L1 on cancer cells and exosomes, and enhances the recognition and killing of cancer cells by human PBMC. Combination of FM213 with the TIGIT inhibitor DTBP-3 (HY-P11903) enhances anti-tumor immunity. FM213 can be used in research related to non-small cell lung cancer.
For research use only. We do not sell to patients.
- Formula: C73H86Cl3N13O12
- Molecular Weight:1443.90
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Storage:
Please store the product under the recommended conditions in the Certificate of Analysis.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
- Protocols
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Description
IC50 & Target
[1]|
PD-L1 323 nM (IC50) |
Cellular Effect
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Cell Line
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Type | Value | Description | References |
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| NCI-H1299 | IC50 |
10 μM
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Inhibition of cell viability against human H1299 NSCLC cells assessed by CCK-8 assay after 48-hour incubation.
Inhibition of cell viability against human H1299 NSCLC cells assessed by CCK-8 assay after 48-hour incubation.
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41729103 |
| H1975 | IC50 |
10 μM
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Inhibition of cell viability against human H1975 NSCLC cells assessed by CCK-8 assay after 48-hour incubation.
Inhibition of cell viability against human H1975 NSCLC cells assessed by CCK-8 assay after 48-hour incubation.
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41729103 |
| HaCaT | IC50 |
10 μM
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Inhibition of cell viability against human HaCaT normal immortalized keratinocyte cells assessed by CCK-8 assay after 48-hour incubation.
Inhibition of cell viability against human HaCaT normal immortalized keratinocyte cells assessed by CCK-8 assay after 48-hour incubation.
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41729103 |
| PBMC | IC50 |
10 μM
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Inhibition of cell viability against human peripheral blood mononuclear cells assessed by CCK-8 assay after 48-hour incubation.
Inhibition of cell viability against human peripheral blood mononuclear cells assessed by CCK-8 assay after 48-hour incubation.
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41729103 |
In Vitro
FM213 (1 h) potently inhibits the interaction between human recombinant PD-1 and PD-L1 proteins in HTRF assays, with an IC50 of 323 nM[1].
FM213 (10 μM) specifically binds to human recombinant PD-L1 and PD-L1 protein, and does not interact with mouse recombinant PD-L1 and PD-1 protein[1].
FM213 (6.25-50 μM) binds to specific surface residues on human recombinant PD-L1 protein, including those at the PD-1 interaction site[1].
FM213 (1-100 μM; 48 h) inhibits the viability of H1299 non-small cell lung cancer cells, H1975 non-small cell lung cancer cells, HaCaT normal keratinocytes, and peripheral blood mononuclear cells (PBMCs)[1].
FM213 (2 μM; 48 h) significantly increases the level of IL-2 and decreases the levels of IL-6 and TNF-α in the co-culture system of H1975 non-small cell lung cancer cells and activated peripheral blood mononuclear cells[1].
FM213 (10 μM; 2 h) thermostabilizes PD-L1 in IFN-γ-stimulated H1299 cells and basal-state H1975 non-small cell lung cancer cells[1].
FM213 (5 μM; 2 h) protects PD-L1 from proteolytic degradation in IFN-γ-stimulated H1299 cells and basal-state H1975 non-small cell lung cancer cells[1].
FM213 (2 μM; 24 h) significantly reduces PD-L1 levels on the surface of IFN-γ-stimulated H1299 cells as well as basal-state H1975 non-small cell lung cancer (NSCLC) cells[1].
FM213 (2 μM; 18-24 h) induces PD-L1 endocytosis and increases its colocalization with lysosomes in IFN-γ-stimulated H1299 cells and basal-state H1975 non-small cell lung cancer (NSCLC) cells[1].
FM213 (2 μM; 24 h) shortens the half-life of PD-L1 in IFN-γ-stimulated H1299 cells and basal-state H1975 non-small cell lung cancer (NSCLC) cells via a lysosome-dependent pathway[1].
FM213 (500 μM) specifically binds to PD-L1 on the surface of ex vivo exosomes derived from A549 non-small cell lung cancer cells[1].
FM213 (2 μM; 48 h) significantly enhances apoptosis in H1299 and H1975 non-small cell lung cancer (NSCLC) cells and increases IFN-γ secretion levels in stimulated peripheral blood mononuclear cells (PBMCs)[1].
The combination of FM213 (2 μM; 48 h) and 2.5 μM DTBP-3 (HY-P11903) synergistically significantly enhances apoptosis in H1299 and H1975 non-small cell lung cancer (NSCLC) cells and increases IFN-γ secretion levels in stimulated peripheral blood mononuclear cells (PBMCs)[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:human NSCLC cell lines (H1299, H1975), normal immortalized human keratinocyte (HaCaT) cells, human peripheral blood mononuclear cells (PBMCs)
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Concentration:1, 2, 5, 10, 25, 50 and 100 μM
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Incubation Time:48 h
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Result:Reduced cell viability in a concentration-dependent manner across all tested cell lines.
The IC50 value for cell viability inhibition was 10 μM in all tested cell lines.
At 2 μM, cell viability remained near 100% in all cell lines, confirming this as a non-cytotoxic concentration.
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Cell Line:cocultures of human NSCLC cell lines (H1299, H1975) with stimulated human PBMCs
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Concentration:2 μM
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Incubation Time:48 h
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Result:Significantly increased the rate of Annexin V-positive apoptotic cells in both H1299 (to ~20%) and H1975 (to ~12%) cell lines compared to untreated cocultures.
Significantly increased IFN-γ secretion by stimulated PBMCs to ~3500 pg/mL in H1299 cocultures and ~3800 pg/mL in H1975 cocultures, which was higher than levels induced by control compounds.
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Cell Line:cocultures of human H1975 NSCLC cells with stimulated human PBMCs
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Concentration:2 μM
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Incubation Time:48 h
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Result:Significantly increased IL-2 levels to a mean of 404.9 pg/mL.
Significantly reduced IL-6 levels to a mean of 3150 pg/mL.
Significantly reduced TNF-α levels to a mean of 30.22 pg/mL compared to untreated cocultures.
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Cell Line:human NSCLC cell lines (H1299, H1975) with IFN-γ-induced PD-L1 upregulation (H1299) or basal PD-L1 expression (H1975)
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Concentration:2 μM
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Incubation Time:18 h, 24 h
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Result:Dramatically reduced cell-surface PD-L1 expression and increased colocalization of PD-L1 with lysosomes after 18 and 24 hours of treatment, indicating PD-L1 internalization and lysosomal targeting.
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Cell Line:human NSCLC cell lines (H1299, H1975) with IFN-γ-induced PD-L1 upregulation (H1299) or basal PD-L1 expression (H1975)
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Concentration:2 μM this compound; 20 μM Cycloheximide (HY-12320), 30 μM Chloroquine (HY-17589A)
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Incubation Time:24 h (this compound); 2 h, 4 h, 6 h (Cycloheximide (HY-12320), Chloroquine (HY-17589A))
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Result:In the presence of cycloheximide, significantly reduced the PD-L1 half-life to less than 6 hours in both cell lines compared to cycloheximide-only controls.
This effect was abrogated by cotreatment with chloroquine, confirming lysosome-dependent degradation.
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Cell Line:cocultures of human NSCLC cell lines (H1299, H1975) with stimulated human PBMCs, in combination with TIGIT inhibitor DTBP-3
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Concentration:2 μM this compound, 2.5 μM DTBP-3 (HY-P11903)
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Incubation Time:48 h
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Result:Cotreatment significantly increased the rate of Annexin V-positive apoptotic cells in both H1299 (to ~50%) and H1975 (to ~60%) cell lines compared to single-agent treatments and untreated cocultures.
Significantly increased IFN-γ secretion by stimulated PBMCs to ~5000 pg/mL in H1299 cocultures and ~5500 pg/mL in H1975 cocultures, which was higher than levels induced by single-agent treatments.
Chemical Information
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Molecular Weight 1443.90
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Formula C73H86Cl3N13O12
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Sequence
Cyclo({Phe(4-Cl)}-Lys-{Phe(4-Cl)}-{d-Pro}-Pro-Trp-Tyr-{Phe(4-Cl)}-Leu-Asn)
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Sequence Shortening
Cyclo({Phe(4-Cl)}-K-{Phe(4-Cl)}-{d-Pro}-PWY-{Phe(4-Cl)}-LN)
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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.
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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Lysosome and acidic-vesicle live-cell staining
Lysosome and acidic-vesicle live-cell staining detects acidic intracellular compartments by using membrane-permeant acidotropic probes that accumulate in low-pH vesicles, including lysosomes, late endosomes, autolysosomes, and acidic phagosomes. LysoTracker staining is commonly used as an intensity-based readout of acidic lysosomal compartment abundance or enlargement, while acridine orange produces green fluorescence in less concentrated compartments and red fluorescence after concentration-dependent accumulation in acidic vesicular organelles. Loss or reduction of acridine-orange red signal can be used as a readout of lysosomal membrane permeabilization or reduced acidic-vesicle integrity. This protocol is designed for live cultured cells and can be adapted for fluorescence microscopy, high-content imaging, plate-reader readout, or flow cytometry when the selected literature supports the readout. Because these dyes report acidotropic accumulation rather than lysosome identity alone,
Purity & Documentation
References
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
FM213 Related Classifications
HY-P11891 Related Classifications
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