PAC-1

Catalog No.S2738 Batch:S273801

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Technical Data

Formula

C23H28N4O2
Molecular Weight 392.49 CAS No. 315183-21-2
Solubility (25°C)* In vitro DMSO 78 mg/mL (198.73 mM)
Water Insoluble
Ethanol Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description PAC-1 is a potent procaspase-3 activator with EC50 of 0.22 μM and the first small molecule known to directly activate procaspase-3 to caspase-3.
Targets
Procaspase-3 [1]
0.22 μM(EC50)
In vitro PAC-1 activates procaspase-7 in a less efficient manner with EC50 of 4.5 μM. Elevated caspase 3 level in cancer cell lines allows PAC-1 to selectively induce apoptosis in a manner proportional to procaspase-3 concentration with IC50 of 0.35 μM for NCI-H226 cells to ~3.5 μM for UACC-62 cells. PAC-1 induces apoptosis in the primary cancerous cells with IC50 values of 3 nM to 1.41 μM, more potently than in the adjacent noncancerous cells with IC50 of 5.02 μM to 9.98 μM, which is also directly related to the distinct procaspase-3 concentration. [1] PAC-1 activates procaspase-3 by chelating zinc ions, thus relieving the zinc-mediated inhibition and allowing procaspase-3 to auto-activate itself to caspase-3. [2] PAC-1 is capable to induce cell death in Bax/Bak double-knockout cells and Bcl-2 and Bcl-xL-overexpressing cells with the same efficacy as its wild-type counterpart in a delayed manner. PAC-1 induces cytochrome c release in a caspase-3 independent manner, which subsequently triggers downstream caspase-3 activation and cell death. PAC-1 can not induce cell death and caspase-3 activation in Apaf-1 knockout cells, suggesting that apoptosome formation is essential for caspase-3 activation by PAC-1-mediated cell death. [3]
In vivo Administration of PAC-1 at 5 mg with low and steady releasing significantly inhibits the growth of ACHN renal cancer xenograft in mice. Oral administration of PAC-1 (50 or 100 mg/kg) significantly retards tumor growth of NCI-H226 lung cancer xenograft in a dose-dependent manner, and markedly prevents the cancer cells from infiltrating the lung tissue. The in vivo anti-tumor effect of PAC-1 is ascribed to procaspase-3 activation and subsequently apoptosis induction consistent with the activity in vitro. [1]
Features The first small molecule known to directly activate procaspase-3 to caspase-3.

Protocol (from reference)

Kinase Assay:[1]
  • In vitro procaspase-3 activation

    Procaspase-3 is expressed and purified in Escherichia coli. Various concentrations of PAC-1 are added to 90 μL of a 50 ng/mL of procaspase-3 in caspase assay buffer in a 96-well plate, The plate is incubated for 12 hours at 37 °C. A 10 μL volume of a 2 mM solution of caspase-3 peptidic substrate acetyl Asp-Glu-Val-Asp-p-nitroanilide (Ac-DEVD-pNa) in caspase assay buffer is then added to each well. The plate is read every 2 minutes at 405 nm for 2 hours in a Spectra Max Plus 384 well plate reader. The slope of the linear portion for each well is determined, and the relative increase in activation from untreated control wells is calculated.
Cell Assay:[1]
  • Cell lines

    U-937, HL-60, CRL-1872, ACHN, NCI-H226, Hs888Lu, Hs578Bst, MCF-10A, SK-MEL-5, BT-20, MDA-MB-231, UACC-62, SK-N-SH, B16-F10 , Hs 578t, and PC-12

  • Concentrations

    Dissolved in DMSO, final concentrations ~100 μM

  • Incubation Time

    72 hours

  • Method

    Cells are exposed to various concentrations of PAC-1 for 72 hours. Cell death is quantified by the addition of MTS/PMS CellTiter 96 Cell Proliferation Assay reagent. The plates are incubated at 37 °C for approximately 1 hour (until the colored product formed), and the absorbance is measured at 490 n
Animal Study:[1]
  • Animal Models

    Ovariectomized female athymic BALB/c (nude, nu/nu) mice injected subcutaneously with ACHN cells, male athymic BALB/c nude mice injected subcutaneously with NCI-H226 cells, and male athymic BALB/c–/– mice injected intravenously with NCI-H226 cells

  • Dosages

    ~100 mg/kg

  • Administration

    Pellet implantation subcutaneously or oral gavage

Customer Product Validation

Data from [Data independently produced by , , Biol Med, 2015, 7(5).doi: 10.4172/0974-8369.1000259.]

Selleck's PAC-1 has been cited by 17 publications

Aedes aegypti Argonaute 2 controls arbovirus infection and host mortality [ Nat Commun, 2023, 14(1):5773] PubMed: 37723154
Aedes aegypti Argonaute 2 controls arbovirus infection and host mortality [ Nat Commun, 2023, 14(1):5773] PubMed: 37723154
Caspase 3 exhibits a yeast metacaspase proteostasis function that protects mitochondria from toxic TDP43 aggregates [ Microb Cell, 2023, 10(8):157-169] PubMed: 37545643
PEBP balances apoptosis and autophagy in whitefly upon arbovirus infection [ Nat Commun, 2022, 13(1):846] PubMed: 35149691
COL8A1 Promotes NSCLC Progression Through IFIT1/IFIT3-Mediated EGFR Activation [ Front Oncol, 2022, 12:707525] PubMed: 35280763
ANP32E contributes to gastric cancer progression via NUF2 upregulation [ Mol Med Rep, 2022, 26(3)275] PubMed: 35795988
Modulating environmental signals to reveal mechanisms and vulnerabilities of cancer persisters [ Sci Adv, 2022, 8(4):eabi7711] PubMed: 35089788
Microbes exploit death-induced nutrient release by gut epithelial cells [ Nature, 2021, 10.1038/s41586-021-03785-9] PubMed: 34349263
The Zebrafish Antiapoptotic Protein BIRC2 Promotes Edwardsiella piscicida Infection by Inhibiting Caspases and Accumulating p53 in a p53 Transcription-Dependent and -Independent Manner [ Front Immunol, 2021, 12:781680] PubMed: 34887869
Cooperative application of transcriptomics and ceRNA hypothesis: LncRNA-107052630/miR-205a/G0S2 crosstalk is involved in ammonia-induced intestinal apoptotic injury in chicken. [ J Hazard Mater, 2020, 18;396:122605] PubMed: 32334290

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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