| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
The compound targets the PD-1 receptor on the surface of activated T cells. By binding to PD-1 or its ligand PD-L1 (the exact binding mode is not specified in standard references), PD1-PDL1-IN 1 TFA prevents the engagement of PD-1 with PD-L1. This blocks the inhibitory signal that normally suppresses T cell activation, allowing T cells to recognize and kill cancer cells more effectively.
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| ln Vitro |
The ~55kD type I membrane glycoprotein known as PD-1, also known as programmed cell death 1 or PDCD1, is a member of the CD28 superfamily of receptors that, by interaction with particular ligands, adversely controls T cell antigen receptor signaling. Maintaining self-tolerance is believed to be a major function of PD-1 [1].
In co-culture assays, PD1-PDL1-IN 1 TFA enhances T cell-mediated killing of tumor cells. T cell proliferation and IFN-gamma secretion increase in the presence of the compound compared to controls with PD-L1-expressing tumor cells alone. It shows dose-dependent activity in functional immune cell assays, with typical EC50 values in the low nanomolar to sub-micromolar range depending on the assay system (specific data not provided in basic references). |
| ln Vivo |
A soluble version of human PD-1 is coated on a 96-well plate (0.5-2 ug/mL in PBS, overnight at 4degC). After blocking with 1% BSA, biotinylated PD-L1 (0.1-0.5 ug/mL) is added to the wells with or without varying concentrations (0.1-1000 nM) of PD1-PDL1-IN 1 TFA. After 2 hours of incubation at room temperature, streptavidin-HRP is added, followed by TMB substrate. The reaction is stopped with 2N H2SO4, and absorbance is read at 450 nm. The IC50 is calculated by curve-fitting. Alternatively, an AlphaScreen or TR-FRET assay can be used to measure PD-1/PD-L1 interaction in a homogeneous format. A PD-1/PD-L1 LANCE Ultra TR-FRET kit (PerkinElmer) is used: 5 nM of biotinylated PD-1, 5 nM of Eu-labeled anti-PD-1 antibody, and 50 nM of Alexa Fluor 647-labeled PD-L1 are mixed with compound (0.01-10 uM) in assay buffer (50 mM HEPES, pH 7.4, 100 mM NaCl, 0.1% BSA, 0.05% Tween-20) in a 384-well plate. After 2 hours at room temperature, the TR-FRET signal (excitation 320 nm, emission 665 nm) is measured on an EnVision plate reader. IC50 is calculated as the compound concentration that reduces the signal by 50%.
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| Enzyme Assay |
Peripheral blood mononuclear cells (PBMCs) are isolated from healthy donors by Ficoll-Paque gradient centrifugation. PBMCs (2×10⁵ cells/well) are seeded in 96-well plates and co-cultured with PD-L1-expressing tumor cells (e.g., MDA-MB-231 breast cancer cells, 2×10⁴ cells/well) in RPMI-1640 + 10% FBS. Cells are treated with PD1-PDL1-IN 1 TFA at concentrations ranging from 46.9 nM to 1500 nM for two hours prior to stimulation. T cell activation is stimulated by adding anti-CD3/CD28 beads or PMA/ionomycin. After 48-72 hours, cell supernatants are collected and IFN-gamma levels are measured by ELISA. T cell proliferation is assessed by CFSE dilution or by measuring Ki67 expression. Tumor cell killing is quantified by LDH release assay or real-time impedance-based monitoring. Appropriate controls include no compound, control vehicle, and a positive control anti-PD-1 antibody. Use of multiple PBMC donors is advised due to inter-donor variability.
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| Cell Assay |
Currently, specific data for PD1-PDL1-IN 1 TFA are not available. For a typical PD-1/PD-L1 inhibitor in a mouse tumor model, the following protocol is representative: 6-8 week old female C57BL/6 mice (n=8-10 per group) are implanted subcutaneously with 5×10⁵ MC38 (colon adenocarcinoma) or B16-F10 (melanoma) cells. When tumors reach 50-100 mm3, mice are randomized and treated with PD1-PDL1-IN 1 TFA (dissolved in 10% DMSO + 40% PEG400 + 50% PBS or sterile saline) administered orally or intraperitoneally at doses of 10-50 mg/kg daily or every other day. Control groups receive vehicle alone or anti-PD-1 antibody (200 microg/mouse, i.p., twice weekly). Tumor volume (width2 × length/2) is measured every 2-3 days for 2-3 weeks. At study termination, tumors are excised, weighed, and analyzed for immune cell infiltration by flow cytometry (CD4+ T cells, CD8+ T cells, regulatory T cells) and cytokine levels (IFN-gamma, TNF-alpha, IL-2). T cell activation markers (CD69, CD25) and PD-1 expression are assessed. Survival is monitored as a secondary endpoint.
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| Animal Protocol |
Pharmacokinetic data (half-life, AUC, bioavailability) are not available for PD1-PDL1-IN 1 TFA. As a TFA salt of a small molecule, the compound is expected to be water-soluble. For in vivo experiments, typical dosages for PD-1/PD-L1 small-molecule inhibitors range from 10-50 mg/kg administered orally or intraperitoneally, with dosing frequency ranging from once daily to every other day. Since specific PK parameters for this compound are not published, pilot PK studies in rodents are recommended.
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| ADME/Pharmacokinetics |
No specific toxicity data have been reported for PD1-PDL1-IN 1 TFA. However, as an immune checkpoint inhibitor, the primary safety concern is immune-related adverse events (irAEs), including autoimmune reactions such as colitis, pneumonitis, dermatitis, and endocrinopathies. These are mechanism-based and reflect the enhanced activity of the immune system. The severity of irAEs is typically dose- and exposure-dependent. In preclinical animal models, high doses may cause weight loss, diarrhea, and skin rashes. Standard animal toxicology studies (acute, sub-chronic) have not been published.
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| Toxicity/Toxicokinetics |
PD1-PDL1-IN 1 TFA is a research-grade compound and is not FDA-approved for clinical use. It is distinct from the macrobicyclic peptide drug pembrolizumab and the small molecule inhibitors like BMS-202. This compound is used in early discovery and validation studies to explore the therapeutic potential of PD-1/PD-L1 blockade. The TFA salt form enhances solubility and handling properties for in vitro assays. Several PD-1/PD-L1 small-molecule inhibitors are in preclinical development and clinical trials, but this specific compound's development status is not available. The compound is stable for 1-2 years when stored as a powder at -20degC. For cell culture, stock solutions (10-100 mM in DMSO) can be stored at -80degC for up to 3 months; avoid freeze-thaw cycles.
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| References |
| Molecular Formula |
C16H24F3N7O8
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|---|---|
| Molecular Weight |
499.40
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| Related CAS # |
PD1-PDL1-IN 1;2005454-12-4
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| Appearance |
Off-white to light yellow solid powder
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 :~100 mg/mL (~200.24 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.01 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 25.0 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: ≥ 2.5 mg/mL (5.01 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (5.01 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0024 mL | 10.0120 mL | 20.0240 mL | |
| 5 mM | 0.4005 mL | 2.0024 mL | 4.0048 mL | |
| 10 mM | 0.2002 mL | 1.0012 mL | 2.0024 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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