Prima-1(also known as PRIMA-1MET) is a mutant p53 reactivator that induces apoptosis and inhibits the growth of human tumors. PRIMA-1, also known as APR-246, is a novel and powerful small molecule compound that is the active form of APR-246. It may be used to treat a variety of p53 mutant-dependent human tumor types. PRIMA-1 exhibits anti-tumor activity in vivo and restores the mutant p53 proteins' capacity to suppress tumor growth. Mutant p53 is highly expressed in many tumors and serves as an essential tumor suppressor. PRIMA-1 is viewed as a potential lead compound for the creation of anticancer medications that target mutant p53.

Physicochemical Properties

Molecular Formula	C9H15NO3
Molecular Weight	185.22
Exact Mass	185.105
Elemental Analysis	C, 58.36; H, 8.16; N, 7.56; O, 25.91
CAS #	5608-24-2
Related CAS #	Eprenetapopt;5291-32-7
PubChem CID	322968
Appearance	White to off-white solid powder
Density	1.3±0.1 g/cm3
Boiling Point	353.7±17.0 °C at 760 mmHg
Flash Point	167.7±20.9 °C
Vapour Pressure	0.0±1.8 mmHg at 25°C
Index of Refraction	1.582
LogP	0.7
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	13
Complexity	217
Defined Atom Stereocenter Count	0
SMILES	O=C1C(CO)(CO)N2CCC1CC2
InChi Key	RFBVBRVVOPAAFS-UHFFFAOYSA-N
InChi Code	InChI=1S/C9H15NO3/c11-5-9(6-12)8(13)7-1-3-10(9)4-2-7/h7,11-12H,1-6H2
Chemical Name	2,2-bis(hydroxymethyl)-1-azabicyclo[2.2.2]octan-3-one
Synonyms	Prima-1
HS Tariff Code	2934.99.9001
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)

Biological Activity

Targets	p53
ln Vitro	PRIMA-1 (NSC-281668) is cultivated with the cell lines at concentrations ranging from 0-140 μM. For PANC-1, HEC-1-B, SUM149, AN 3CA, Ishikawa, Panc02, and MDA-MB-231 cells, respectively, the IC50s are 35, 40, 50, 50, 60, 70, and 75 μM.
ln Vivo	PRIMA-1 (Prima-1) is a substance that modifies p53. After 17 weeks and 34 weeks, respectively, 150 or 300 ppm PRIMA-1 significantly reduces ((P<0.0001) the development of lung adenocarcinomas by 56% and 62%, respectively. After 17 weeks of exposure and 34 weeks of exposure, administration of 150 or 300 ppm PRIMA-1 significantly reduces NNK-induced total lung adenocarcinoma formation by 57% or 62% (P<0.0001), respectively, and by 39% or 56% (P<0.0001). The number of NNK-induced lung adenomas is slightly increased when the lower (150 ppm) dose of PRIMA-1 is administered, just as it is when the lower (50 ppm) dose of CP-31398 is administered[3].
Enzyme Assay	PRIMA-1 is a mutant p53 reactivator, restores the sensitivity of TP53 mutant-type thyroid cancer cells to the histone methylation inhibitor 3-Deazaneplanocin A.
Cell Assay	A cell viability assay using yellow tetrazolium salt3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide or MTT is utilized to assess the effects of the p53 SMWC on growth of human carcinoma cell lines. At a density of 2.5×103 cells per well in 100 L of complete medium, cells are plated in triplicate in 96-well plates. The cells are treated with increasing concentrations of SMWC for an additional 24 hours, during which time they are incubated at 37 degrees Celsius in a humidified 5% atmosphere. The cells are then examined for cell growth using the MTT assay. PrIMA-1 and CP-31398 stock solutions (10 mM each) are diluted in PBS just before use. The assay is carried out as follows: a 12 mM MTT stock solution is made by combining 5 mg MTT with 1 mL of sterile PBS and vortexing or sonicating the mixture until the MTT is completely dissolved. After being prepared, the MTT solution is kept at 4°C in a light-protected environment for four weeks. The mixture is gently inverted until the SDS dissolves to create a 500 mL SDS-HCl solution, which contains 0.01 M HCl, 10% propanol, and 5 gm SDS. Each well receives 10 μL of the 12 mM MTT stock solution after 100 μL of cell culture medium have been removed. Prepare a negative control by mixing 10 μL of the MTT stock solution with 100 μL of medium.
Animal Protocol	Dissolved in PBS; 1, 10, 20 and 100 mg/kg; i.v. injection SCID mice
References	[1]. PRIMA-1, a mutant p53 reactivator, restores the sensitivity of TP53 mutant-type thyroid cancer cells to the histone methylation inhibitor 3-Deazaneplanocin A. J Clin Endocrinol Metab. 2014 Jun;99(6):E962-70. [2]. Targeting cancer stem cells with p53 modulators. Oncotarget. 2016 Apr 8. [3]. Chemopreventive effects of the p53-modulating agents CP-31398 and Prima-1 in tobacco carcinogen-induced lung tumorigenesis in A/J mice. Neoplasia. 2013 Sep;15(9):1018-27.
Additional Infomation	2,2-bis(hydroxymethyl)-1-azabicyclo[2.2.2]octan-3-one is a member of quinuclidines and a cyclic ketone.

Solubility Data

Solubility (In Vitro)

DMSO: ~37 mg/mL (~199.8 mM) Water: ~37 mg/mL (~199.8 mM) Ethanol: N/A

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (13.50 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 (13.50 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (13.50 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly. Solubility in Formulation 4: Prima 1;Prima1;Prima-1 Solubility in Formulation 5: 50 mg/mL (269.95 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	5.3990 mL	26.9949 mL	53.9898 mL
	5 mM	1.0798 mL	5.3990 mL	10.7980 mL
	10 mM	0.5399 mL	2.6995 mL	5.3990 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.