| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Perforin-IN-2 directly targets the pore-forming protein perforin, which is a key effector molecule stored in the cytotoxic granules of CD8+ T lymphocytes and natural killer cells. Perforin plays an essential role in the immune system by forming transmembrane pores in the plasma membrane of target cells, leading to osmotic lysis and cell death. By inhibiting perforin, Perforin-IN-2 blocks the cytolytic activity of these immune cells without affecting other immune functions, thereby providing a targeted approach to immunosuppression in the context of transplantation.
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| ln Vitro |
Perforin-IN-2 effectively inhibits the lytic activity of recombinant perforin on Jurkat T leukemia cells with an IC50 of 6.65 μM. It also prevents the death of K562 leukemia target cells, demonstrating its ability to block perforin-mediated cytotoxicity in a cellular context. The compound binds to plasma proteins with an exceptionally high binding rate of 99.8%, which significantly influences its distribution and bioavailability. In enzymatic assays, Perforin-IN-2 shows concentration-dependent inhibition of perforin pore-forming activity, validating its mechanism of action as a direct perforin inhibitor.
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| ln Vivo |
In vivo, Perforin-IN-2 requires a relatively high concentration (>900 μM) to achieve optimal perforin inhibition, indicating that significant compound accumulation at the target site may be necessary for therapeutic efficacy. The compound has been evaluated in preclinical models of allogeneic transplantation, where it alleviates transplant rejection by reducing perforin-mediated killing of donor cells. Its high plasma protein binding (99.8%) suggests that the free, active concentration of the compound in circulation is extremely low, which may necessitate high total drug concentrations to achieve sufficient target engagement in vivo.
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| Enzyme Assay |
Non-cell-based assays for Perforin-IN-2 typically involve biochemical binding and inhibition studies using purified recombinant perforin protein. A standard protocol includes incubating recombinant perforin with various concentrations of the compound (ranging from 0.1 to 100 μM) in a suitable buffer system, followed by the addition of a fluorogenic or colorimetric substrate to measure residual perforin activity. The inhibition of perforin-mediated liposome disruption or erythrocyte lysis can be quantified spectrophotometrically to determine the IC50 value. Surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC) may be employed to directly measure the binding affinity (Kd) between Perforin-IN-2 and perforin, providing mechanistic insights into the inhibition kinetics.
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| Cell Assay |
Cell-based assays for Perforin-IN-2 typically utilize perforin-sensitive target cell lines such as Jurkat T leukemia cells or K562 erythroleukemia cells. A representative protocol involves culturing target cells in appropriate medium, labeling them with a fluorescent dye such as calcein-AM or using a chromium-51 release assay, and co-incubating them with recombinant perforin or effector cells in the presence of varying concentrations of Perforin-IN-2 (typically 0.1–100 μM). After incubation at 37°C for 2–4 hours, cell lysis is quantified by measuring the release of fluorescent or radioactive labels into the supernatant. The percentage of inhibition is calculated relative to control wells without compound, and dose-response curves are generated to determine the IC50 for cytolysis inhibition.
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| Animal Protocol |
In vivo animal studies with Perforin-IN-2 are conducted in murine models of allogeneic bone marrow or stem cell transplantation. A typical protocol involves conditioning recipient mice with irradiation or chemotherapy, followed by transplantation of donor bone marrow cells. Perforin-IN-2 is administered via intraperitoneal or oral routes at doses ranging from 10 to 100 mg/kg, starting before transplantation and continuing for several days or weeks post-transplant. Graft survival, chimerism levels, and signs of graft-versus-host disease are monitored. Pharmacodynamic endpoints include assessment of perforin activity in isolated cytotoxic lymphocytes and histopathological examination of target organs such as skin, liver, and intestine.
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| ADME/Pharmacokinetics |
Detailed pharmacokinetic parameters of Perforin-IN-2 have been characterized in preclinical studies published in ACS Pharmacology & Translational Science. The compound exhibits an exceptionally high plasma protein binding rate of 99.8%, resulting in a very low fraction of unbound drug available for tissue distribution and target engagement. This pharmacokinetic property necessitates high total drug concentrations (>900 μM) to achieve optimal perforin inhibition in vivo. The compound is typically administered via injection in research settings, with bioavailability and half-life data available from the primary literature. The high protein binding may also influence the compound's volume of distribution and clearance profile.
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| Toxicity/Toxicokinetics |
Toxicological data for Perforin-IN-2 are derived from preclinical studies and indicate that the compound is intended for research use only and is not approved for human therapeutic applications. The high concentration required for in vivo efficacy (>900 μM) raises potential concerns about the therapeutic window and off-target effects. Standard safety assessments would include cytotoxicity screening, hERG channel inhibition testing, and preliminary toxicology studies in animal models to evaluate maximum tolerated dose and target organ toxicity. As a research compound, appropriate safety precautions should be followed during handling, including the use of personal protective equipment and proper chemical hygiene practices.
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| References | |
| Additional Infomation |
Perforin-IN-2 (Compound 1) is a first-in-class benzosulfonamide perforin inhibitor that has been validated in preclinical models of allogeneic transplantation. The compound is referenced in the primary literature, specifically in a study published in ACS Pharmacology & Translational Science (2022) that describes the preclinical activity and pharmacokinetic/pharmacodynamic relationship for a series of novel benzenesulfonamide perforin inhibitors. Perforin-IN-2 represents a promising tool compound for studying the role of perforin in immune-mediated diseases beyond transplantation, including autoimmune disorders and certain cancers where perforin-mediated cytotoxicity plays a pathogenic role. The compound is stored as a powder at -20°C for up to 3 years and is soluble in DMSO at 33.33 mg/mL.
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| Exact Mass |
506.072
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| CAS # |
1567836-70-7
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| PubChem CID |
73295278
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| Appearance |
Light yellow to yellow solid powder
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
35
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| Complexity |
907
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1CC2=C(C1=O)C=CC(=C2)C3=CC=C(S3)C4=CC(=CN=C4)NS(=O)(=O)C5=CC=CC=C5[N+](=O)[O-]
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| InChi Key |
BIAQRTAXQYZHBE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C24H18N4O5S2/c1-27-14-17-10-15(6-7-19(17)24(27)29)21-8-9-22(34-21)16-11-18(13-25-12-16)26-35(32,33)23-5-3-2-4-20(23)28(30)31/h2-13,26H,14H2,1H3
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| Chemical Name |
N-[5-[5-(2-methyl-1-oxo-3H-isoindol-5-yl)thiophen-2-yl]-3-pyridinyl]-2-nitrobenzenesulfonamide
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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 |
| 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 :~33.33 mg/mL (~65.80 mM; with sonication (<80°C))
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| Solubility (In Vivo) |
Solubility in Formulation 1: 2.5 mg/mL (4.94 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one),clear solution; with ultrasonication.
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 and add it to 900 μL corn oil and mix well. (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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