1149-23-1 Purity
95%
If you have any other questions or need other size, please get a quote.
Specification
TP53 mutations that impair p53 transcriptional function occur in more than 50% of human cancers, including pancreatic adenocarcinoma, and render cancer cells more resistant to conventional treatments. In the past few years, many efforts have been made to identify p53 reactivating molecules that can restore the mutant protein to its wild-type transcriptionally active conformation. Two of these compounds, CP-31398 dihydrochloride hydrate and RITA, induced cell growth inhibition, apoptosis, and autophagy by activating p53/DNA binding and p53 phosphorylation (Ser15) without affecting the total amount of p53. These effects occurred in both wild-type and mutant p53 pancreatic adenocarcinoma cell lines and were less pronounced in normal human primary fibroblasts. Furthermore, CP-31398 dihydrochloride hydrate and RITA regulated the SESN12/AMPK/mTOR axis by inducing AMPK phosphorylation on Thr172, which plays a crucial role in the autophagic response.
CP-31398 dihydrochloride hydrate was dissolved in sterile water. RITA [5,5-(2,5-furandiyl)bis-2-thiophenecarbinol; Reactivation of p53 and induction of tumor cell apoptosis was dissolved in DMSO. PaCa3 and Panc1 human pancreatic adenocarcinoma cell lines harboring wild-type and R273H mutant p53, respectively, were grown in RPMI 1640 supplemented with 2 mM glutamine, 10% FBS, and 50 lg/ml. Normal primary fibroblasts and non-small cell lung cancer cell lines were grown in DMEM supplemented with 2 mM glutamine, 10% FBS, and 50 lg/ml gentamicin sulfate. All cell lines were incubated at 37°C, 5% CO .
In the past few years, screening approaches have led to the identification of small molecules, such as CP-31398 dihydrochloride hydrate and RITA, that can reconstitute the wild-type transcriptionally active conformation of mutant p53 proteins. These molecules are able to activate wild-type-like p53 functions in cancer cells expressing the mutant protein, triggering p53-dependent tumor suppression. Treatment with the p53 reactivators CP-31398 dihydrochloride hydrate or RITA reduced ROS levels in cancer cells with wild-type p53 (PaCa3) and mutant p53 (Panc1); whereas ROS levels in p53-deficient AsPC1 cells were not affected by these compounds, indicating that p53 expression is required for their effects. These results confirm the antioxidant role of wild-type p53 and further suggest that mutp53-dependent prooxidant functions can be restored by activating the wild-type-like conformation of the protein.
Gemcitabine (2',2'-difluoro-2'-deoxycytidine) was dissolved in sterile double-distilled water. N-acetyl-β-cysteine (NAC) and CP-31398 dihydrochloride hydrate were dissolved in double-distilled sterile water. 5-aminoimidazole-4-carboxamide ribonucleotide (AICA-R) was dissolved in double-distilled sterile water. RITA 5,5'-(2,5-furandiyl)bis-2-thiophenecarbinol; Reactivation of p53 and Induction of Tumor Cell Apoptosis was dissolved in DMSO. Cells were seeded into 96-well plates and incubated with various compounds or transfected with the indicated constructs the next day under the indicated conditions. At the end of treatment, cell growth was measured by crystal violet assay according to the manufacturer's protocol and absorbance was measured by spectrophotometric analysis (A 595 nm).
CP-31398 is a compound that can inhibit excitotoxicity-induced neurodegeneration. It can rescue the structure and function of the tumor suppressor protein p53 mutant and stabilize the active conformation of p53 wild type, thereby inhibiting axonal degeneration and cell body death caused by excitotoxicity.
Experimental methods and results
· Hippocampal cultures subjected for neurite beading analysis were incubated for 8 DIV at 37 °C, 5% CO2, and treated with 50 μM glutamate, 10 μM CP-31398, or vehicle.
· CP-31398 inhibited more than 90% of the excitotoxicity-induced axonal degeneration phenotype compared to glutamate treatment alone, and CP-31398 treatment alone had no significant effect on axonal integrity. This result suggests that CP-31398 effectively protects axons from excitotoxicity-induced degeneration.
· CP-31398 effectively protected cell bodies from excitotoxicity-induced death, but to a lesser extent compared to the degree of protection observed in axons.
· An increase in mitochondrial function of more than 80% was observed with CP-31398 treatment compared to glutamate treatment alone. Under normal conditions, a modest decrease in mitochondrial dehydrogenase activity was detected after CP-31398 treatment. This result suggests that CP-31398 effectively protects mitochondria from dysfunction caused by excitotoxicity.
In this work, the effects of gemcitabine (GEM) in combination with CP-31398 and RITA were analyzed on pancreatic ductal adenocarcinoma (PDAC) cell lines expressing wt or mp53. The findings indicated that even though gemcitabine had a negative impact on the activation of mutant p53, treating PDAC cells with gemcitabine along with p53 reactivating compounds (CP-31398 and RITA) resulted in decreased growth rate and apoptosis.
· In order to combat chemoresistance to gemcitabine (GEM) treatment in pancreatic ductal adenocarcinoma (PDAC) cells expressing mutant p53 protein (mp53), the p53 reactivating compound CP-31398 was used and the expression of CdK1 and CCNB1 genes was evaluated. CP-31398 has the ability to stabilize a wild-type-like conformation of mutant p53 proteins. Treating Panc1 cells with a combination of CP-31398 and GEM led to a significant suppression of mp53 target gene expression compared to cells treated with GEM alone. This drug combination notably decreased the expression of Cdk1 and CCNB1 genes in PaCa3 cells compared to treatment with GEM or CP-31398 alone.
· In addition, p53 and phosphorylated p53 (P-p53) protein expression after treatment with GEM alone or in combination with CP-31398 was examined in mutant and wild-type p53 cell lines. The findings revealed that cells treated with both GEM and CP-31398 exhibited an elevation in the phosphorylation of Ser15 residue of p53 when compared to untreated or singly treated cells. However, the total expression of p53 protein remained unaffected by these treatments.
The molecular formula is C22H28Cl2N4O.
The molecular weight is 435.4 g/mol.
The IUPAC name is N-[2-[(E)-2-(4-methoxyphenyl)ethenyl]quinazolin-4-yl]-N',N'-dimethylpropane-1,3-diamine;dihydrochloride.
The InChIKey is WWIFDJIOGCGSBS-IVKCLRODSA-N.
The canonical SMILES is CN(C)CCCNC1=NC(=NC2=CC=CC=C21)C=CC3=CC=C(C=C3)OC.Cl.Cl.
The CAS number is 1217195-61-3.
The hydrogen bond donor count is 3.
The hydrogen bond acceptor count is 5.
CP-31398 Dihydrochloride Hydrate has 8 rotatable bonds.
The topological polar surface area is 50.3 Ų.