KPT-6566 is a selective, covalent/irreversible inhibitor of prolyl isomerase PIN1. It covalently binds to the catalytic site of PIN1 and selectively inhibits and degrades PIN1. KPT-6566 has IC50 of 640 nM and Ki of 625.2 nM for PIN1 PPIase domain. KPT-6566 may be used in cancer-related research.

Physicochemical Properties

Molecular Formula	C22H21NO5S2
Molecular Weight	443.535843610764
Exact Mass	443.086
CAS #	881487-77-0
PubChem CID	6022998
Appearance	White to yellow solid powder
LogP	4.8
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	30
Complexity	840
Defined Atom Stereocenter Count	0
SMILES	O=C(CSC1C(=O)C2C(=CC=CC=2)C(=NS(C2C=CC(C(C)(C)C)=CC=2)(=O)=O)C=1)O
InChi Key	BXWWOKYIKNEEHJ-PTGBLXJZSA-N
InChi Code	InChI=1S/C22H21NO5S2/c1-22(2,3)14-8-10-15(11-9-14)30(27,28)23-18-12-19(29-13-20(24)25)21(26)17-7-5-4-6-16(17)18/h4-12H,13H2,1-3H3,(H,24,25)/b23-18+
Chemical Name	2-[(4E)-4-(4-tert-butylphenyl)sulfonylimino-1-oxonaphthalen-2-yl]sulfanylacetic acid
Synonyms	KPT6566; KPT 6566; KPT-6566
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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.(2). Please store this product in a sealed and protected environment (e.g. under nitrogen), 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)

Biological Activity

ln Vitro	KPT-6566 (1–5 μM; 0–8 days) suppresses the proliferation of WT fibroblasts [1]. In a PIN1-dependent way, KPT-6566 (0-10 μM; 48 hours) reduces the viability of cancer cells and normal breast epithelial cells [1]. Cyclin D1, PIN1 concentrations, and hyperphosphorylated pRB levels are all impacted by KPT-6566 (0-10 μM; 48 hours) [1]. The mut-p53, NOTCH1, and NRF2 pathways are inhibited by KPT-6566 (2.5–5 μM; 48 hours) [1]. In a PIN1-dependent way, KPT-6566 (0-5 μM; 48 hours) causes damage to DNA [1].
ln Vivo	Mice are not harmful to KPT-6566 (5 mg/kg; intraperitoneally administered once daily for 26 days) [1].
Cell Assay	Cell proliferation assay[1] Cell Types: WT and Pin1 KO mouse embryo-derived immortalized fibroblasts Tested Concentrations: 1 and 5 μM Incubation Duration: 0-8 days Experimental Results: Dose-dependent inhibition of proliferation of WT fibroblasts, whereas Pin1 KO had no effect on fibroblasts. Cell viability assay[1] Cell Types: MCF10A, HMEC, HeLa, LNCaP, SKOV-3, PANC-1, PC-3, MDA-MB-468 and MDA-MB-231 Cell Tested Concentrations: 0-10 μM Incubation Duration: 48 hour Experimental Results: Inhibited the viability of normal breast epithelial cells and cancer cells even at low concentrations, and increased the concentration of PIN1 in MDA-MB-468, SKOV-3, PC-3, LNCaP and PANC-1. Western Blot Analysis[1] Cell Types: Immortalized fibroblasts derived from WT and Pin1 KO mouse embryos and PIN1 KO MDA-MB-231 cells Tested Concentrations: 0-10 μM Incubation Duration: 48 hrs (hours) Experimental Results: Hyperphosphorylated pRB and Cyclin D1 levels, dose diminished - and promoted PIN1 degradation over time. Western Blot Analysis[1] Cell Types: MDA-MB-231, MCF10A, MDA-MB-468, and
Animal Protocol	Animal/Disease Models: 6weeks old female mice, injected with 1 million MDA-MB-231Luc6 cells [1] Doses: 5 mg/kg Route of Administration: intraperitoneal (ip) injection; 5 mg/kg, one time/day; for 26 days Experimental Results:Passed Postmortem morphological analysis revealed no evidence of local or systemic and organ toxicity.
References	[1]. A covalent PIN1 inhibitor selectively targets cancer cells by a dual mechanism of action. Nat Commun. 2017 Jun 9;8:15772.

Solubility Data

Solubility (In Vitro)

DMSO : ~19.23 mg/mL (~43.36 mM)

Solubility (In Vivo)

Solubility in Formulation 1: 1.92 mg/mL (4.33 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 19.2 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: ≥ 1 mg/mL (2.25 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 10.0 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.2546 mL	11.2729 mL	22.5459 mL
	5 mM	0.4509 mL	2.2546 mL	4.5092 mL
	10 mM	0.2255 mL	1.1273 mL	2.2546 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.