| Targets |
HDM‑2 (human double minute 2 protein), specifically the membrane‑bound form on cancer cells. PNC-27 binds to HDM‑2, forming pores in the cancer cell membrane, which leads to osmotic lysis and cell death.
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| ln Vitro |
PNC-27 acetate (50 μg/mL; 0-3 hours) induces cancer cell death[1]. PNC-27 acetate (50 μg/mL; 15 minutes) binds to cell membrane-bound HDM-2 in A2058 and MCF-7 cells[1].
In vitro, PNC-27 acetate (50 ug/mL, 0‑3 hours) rapidly induces cancer cell death. It binds to cell membrane‑bound HDM-2 in A2058 melanoma and MCF‑7 breast cancer cells within 15 minutes. The peptide has been shown to induce lysis of a wide variety of human cancer cells in a time‑ and concentration‑dependent manner, with no significant toxicity to normal human cells. It is also effective against leukemia cell lines. |
| ln Vivo |
PNC-27 (ip; 40 mg/kg; daily; 2-3 weeks) acetate shows antileukemic activity in mice [2].
In vivo, PNC-27 acetate has demonstrated anti‑tumor activity in mouse xenograft models of human cancers, including melanoma and pancreatic cancer. It reduces tumor growth and induces tumor necrosis, with minimal systemic toxicity. It is being researched for acute myeloid leukemia (AML) and other malignancies, with the ability to kill drug‑resistant cancer cells. |
| Enzyme Assay |
A general cell‑free protocol for assessing membrane pore formation is not established. Binding to HDM‑2 can be assessed using a fluorescence polarization assay. Recombinant HDM‑2 protein is incubated with a fluorescently labeled p53 peptide (FAM‑p53) in a binding buffer. Varying concentrations of PNC-27 acetate are added to compete for HDM‑2 binding, and the decrease in fluorescence polarization is measured to calculate the IC50.
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| Cell Assay |
Cell Cytotoxicity Assay[1]
Cell Types: MIA-PaCa-2 Tested Concentrations: 50 μg/mL Incubation Duration: 0-3 hours Experimental Results: Induced 100% cell death in 90 min. A general cellular protocol for PNC-27 acetate: Cancer cells (e.g., A2058 melanoma, MCF‑7 breast cancer, or AML cell lines) are seeded in 96‑well plates. Cells are treated with PNC-27 acetate at concentrations of 10, 25, 50, 100, and 200 ug/mL for 1, 2, 3, 4, and 6 hours. Cell viability is assessed using the MTT assay or trypan blue exclusion. For mechanism studies, cells are treated with 50 ug/mL PNC-27 acetate for 15, 30, 60, and 120 minutes, and HDM‑2 binding is assessed by flow cytometry using an anti‑HDM‑2 antibody. Membrane pore formation is assessed by measuring lactate dehydrogenase (LDH) release. |
| Animal Protocol |
Animal/Disease Models:MllPTD/WT/Flt3ITD/ITD AML mice[2]
Doses: 40 mg/kg Route of Administration: Intraperitoneal injection; 40 mg/kg; once daily; 2 or 3 weeks Experimental Results: Reduced AML engraftment and prolonged survival. General animal protocol for PNC-27 acetate: Female NOD/SCID mice are subcutaneously implanted with 5×10⁶ A2058 melanoma cells or other cancer cells. When tumors reach ~150 mm3, mice are randomized (n=8/group). PNC-27 acetate is formulated in sterile saline or PBS and administered via intraperitoneal (IP) or intratumoral (IT) injection at doses of 10, 20, and 40 mg/kg daily or every other day for 14‑21 days. Tumor volume is measured twice weekly. At the end of the study, tumors are harvested for histopathology (H&E, necrosis), IHC (HDM‑2, Ki‑67), and TUNEL staining. Blood is collected for serum chemistry (LDH) and hematology. |
| ADME/Pharmacokinetics |
General PK protocol for PNC-27 acetate: The peptide is administered to mice via IP (10 mg/kg) and IV (2 mg/kg). Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours. Plasma concentrations are quantified by LC‑MS/MS. PK parameters (Cmax, Tmax, AUC, t1/2, clearance, Vd, and IP bioavailability) are calculated.
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| Toxicity/Toxicokinetics |
General toxicity protocol for PNC-27 acetate: A 14‑day repeat‑dose IP toxicity study is performed in ICR mice. PNC-27 acetate is administered at doses of 10, 30, and 60 mg/kg/day. Parameters include clinical signs, body weight, food consumption, hematology (CBC, differential), serum chemistry (LDH, ALT, AST, BUN, creatinine), and histopathology of major organs (liver, kidney, spleen, heart, lung, pancreas).
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| References |
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| Additional Infomation |
PNC-27 acetate has the molecular formula C188H293N53O44S·xC2H2O2 and a molecular weight of approximately 4031.73 g/mol (free base). The compound is stored as a solid at ‑80degC. PNC-27 selectively targets and kills cancer cells that express membrane HDM‑2. It has been shown to be effective in killing cancer cells from a wide variety of solid tumors and hematologic malignancies, including AML, and drug‑resistant tumors.
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| Appearance |
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: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 : 25 mg/mL
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (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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