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Nutlin-3a MDM2 inhibitor

Name: Nutlin-3a
Brand: Selleck Chemicals
SKU: S8059
Availability: InStock
Rating: 5 (116 reviews)

Cat.No.S8059

Nutlin-3a ((-)-Nutlin-3), the active enantiomer of Nutlin-3, inhibits the p53/MDM2 interaction with IC50 of 90 nM in a cell-free assay. Nutlin-3a induces autophagy and apoptosis in a p53-dependent manner.

Chemical Structure

Molecular Weight: 581.49

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Quality Control

Batch: Purity: 99.79%

99.79

Related Targets	Bcl-2 Caspase PD-1/PD-L1 Ferroptosis p53 Apoptosis related Synthetic Lethality STAT TNF-alpha Ras
Other MDM2/MDMX Inhibitors	Nutlin-3 RG-7112 Brigimadlin Idasanutlin (RG7388) SAR405838 NSC 207895 NVP-CGM097 Siremadlin (HDM201) Nutlin-3b MX69

Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
MV-4-11	Growth Inhibition Assay			IC50=0.61237 μM	SANGER
H4	Growth Inhibition Assay			IC50=0.6628 μM	SANGER
PA-1	Growth Inhibition Assay			IC50=0.87096 μM	SANGER
NKM-1	Growth Inhibition Assay			IC50=1.04931 μM	SANGER
NEC8	Growth Inhibition Assay			IC50=1.21572 μM	SANGER
EoL-1-cell	Growth Inhibition Assay			IC50=1.26701 μM	SANGER
K5	Growth Inhibition Assay			IC50=1.42072 μM	SANGER
QIMR-WIL	Growth Inhibition Assay			IC50=1.60854 μM	SANGER
MOLT-16	Growth Inhibition Assay			IC50=1.78604 μM	SANGER
CHP-212	Growth Inhibition Assay			IC50=1.81369 μM	SANGER
CTB-1	Growth Inhibition Assay			IC50=2.02246 μM	SANGER
MOLT-4	Growth Inhibition Assay			IC50=2.32853 μM	SANGER
A101D	Growth Inhibition Assay			IC50=2.3501 μM	SANGER
DOHH-2	Growth Inhibition Assay			IC50=2.42279 μM	SANGER
ES4	Growth Inhibition Assay			IC50=2.43155 μM	SANGER
SW780	Growth Inhibition Assay			IC50=2.50883 μM	SANGER
VA-ES-BJ	Growth Inhibition Assay			IC50=2.5411 μM	SANGER
RPMI-8866	Growth Inhibition Assay			IC50=2.56214 μM	SANGER
ML-2	Growth Inhibition Assay			IC50=2.56576 μM	SANGER
MSTO-211H	Growth Inhibition Assay			IC50=2.57451 μM	SANGER
JVM-3	Growth Inhibition Assay			IC50=2.59324 μM	SANGER
A3-KAW	Growth Inhibition Assay			IC50=2.61818 μM	SANGER
DK-MG	Growth Inhibition Assay			IC50=2.62471 μM	SANGER
LNCaP-Clone-FGC	Growth Inhibition Assay			IC50=2.64318 μM	SANGER
HT-144	Growth Inhibition Assay			IC50=2.64577 μM	SANGER
NB69	Growth Inhibition Assay			IC50=2.65334 μM	SANGER
A172	Growth Inhibition Assay			IC50=2.67588 μM	SANGER
RS4-11	Growth Inhibition Assay			IC50=2.72407 μM	SANGER
DU-4475	Growth Inhibition Assay			IC50=2.79502 μM	SANGER
SJSA-1	Growth Inhibition Assay			IC50=2.82556 μM	SANGER
BV-173	Growth Inhibition Assay			IC50=2.84439 μM	SANGER
U-2-OS	Growth Inhibition Assay			IC50=2.9107 μM	SANGER
CHP-134	Growth Inhibition Assay			IC50=2.93882 μM	SANGER
D-502MG	Growth Inhibition Assay			IC50=2.97154 μM	SANGER
KS-1	Growth Inhibition Assay			IC50=3.01623 μM	SANGER
A204	Growth Inhibition Assay			IC50=3.05588 μM	SANGER
KGN	Growth Inhibition Assay			IC50=3.08496 μM	SANGER
NCI-H292	Growth Inhibition Assay			IC50=3.12028 μM	SANGER
CAKI-1	Growth Inhibition Assay			IC50=3.12694 μM	SANGER
C2BBe1	Growth Inhibition Assay			IC50=3.17026 μM	SANGER
NB10	Growth Inhibition Assay			IC50=3.20966 μM	SANGER
MHH-NB-11	Growth Inhibition Assay			IC50=3.26827 μM	SANGER
NCI-SNU-1	Growth Inhibition Assay			IC50=3.27843 μM	SANGER
HCT-116	Growth Inhibition Assay			IC50=3.30335 μM	SANGER
G-401	Growth Inhibition Assay			IC50=3.36322 μM	SANGER
MN-60	Growth Inhibition Assay			IC50=3.44092 μM	SANGER
SW982	Growth Inhibition Assay			IC50=3.50848 μM	SANGER
RKO	Growth Inhibition Assay			IC50=3.53936 μM	SANGER
D-283MED	Growth Inhibition Assay			IC50=3.57986 μM	SANGER
LB996-RCC	Growth Inhibition Assay			IC50=3.62558 μM	SANGER
A549	Growth Inhibition Assay			IC50=3.63552 μM	SANGER
LB2241-RCC	Growth Inhibition Assay			IC50=3.65708 μM	SANGER
SK-HEP-1	Growth Inhibition Assay			IC50=3.74297 μM	SANGER
G-402	Growth Inhibition Assay			IC50=3.80832 μM	SANGER
GOTO	Growth Inhibition Assay			IC50=3.84333 μM	SANGER
LOXIMVI	Growth Inhibition Assay			IC50=3.85675 μM	SANGER
NH-12	Growth Inhibition Assay			IC50=4.01959 μM	SANGER
CTV-1	Growth Inhibition Assay			IC50=4.07973 μM	SANGER
CP50-MEL-B	Growth Inhibition Assay			IC50=4.24392 μM	SANGER
RH-18	Growth Inhibition Assay			IC50=4.27706 μM	SANGER
NB17	Growth Inhibition Assay			IC50=4.31768 μM	SANGER
A375	Growth Inhibition Assay			IC50=4.33524 μM	SANGER
IST-MES1	Growth Inhibition Assay			IC50=4.41411 μM	SANGER
MZ2-MEL	Growth Inhibition Assay			IC50=4.50155 μM	SANGER
CAL-54	Growth Inhibition Assay			IC50=4.53019 μM	SANGER
NCI-H28	Growth Inhibition Assay			IC50=4.62717 μM	SANGER
D-247MG	Growth Inhibition Assay			IC50=4.75072 μM	SANGER
NCI-H460	Growth Inhibition Assay			IC50=4.91127 μM	SANGER
MCF7	Growth Inhibition Assay			IC50=5.44244 μM	SANGER
697	Growth Inhibition Assay			IC50=5.4455 μM	SANGER
ONS-76	Growth Inhibition Assay			IC50=5.57009 μM	SANGER
C32	Growth Inhibition Assay			IC50=5.60029 μM	SANGER
OS-RC-2	Growth Inhibition Assay			IC50=5.73887 μM	SANGER
MEL-HO	Growth Inhibition Assay			IC50=5.85667 μM	SANGER
LoVo	Growth Inhibition Assay			IC50=6.01624 μM	SANGER
AGS	Growth Inhibition Assay			IC50=6.14828 μM	SANGER
GI-ME-N	Growth Inhibition Assay			IC50=6.22421 μM	SANGER
H-EMC-SS	Growth Inhibition Assay			IC50=6.386 μM	SANGER
RVH-421	Growth Inhibition Assay			IC50=6.42428 μM	SANGER
SW954	Growth Inhibition Assay			IC50=6.55572 μM	SANGER
NB5	Growth Inhibition Assay			IC50=6.56183 μM	SANGER
NCI-H2122	Growth Inhibition Assay			IC50=6.58793 μM	SANGER
AM-38	Growth Inhibition Assay			IC50=6.75639 μM	SANGER
KNS-81-FD	Growth Inhibition Assay			IC50=6.76494 μM	SANGER
LS-513	Growth Inhibition Assay			IC50=6.79026 μM	SANGER
A427	Growth Inhibition Assay			IC50=6.87829 μM	SANGER
WM-115	Growth Inhibition Assay			IC50=6.9323 μM	SANGER
COLO-829	Growth Inhibition Assay			IC50=7.24188 μM	SANGER
NCI-H1650	Growth Inhibition Assay			IC50=7.39228 μM	SANGER
NCI-H358	Growth Inhibition Assay			IC50=7.44879 μM	SANGER
HT-1080	Growth Inhibition Assay			IC50=7.48254 μM	SANGER
HCC2218	Growth Inhibition Assay			IC50=7.6297 μM	SANGER
NCI-H661	Growth Inhibition Assay			IC50=7.87069 μM	SANGER
KM-H2	Growth Inhibition Assay			IC50=7.88694 μM	SANGER
RPMI-2650	Growth Inhibition Assay			IC50=7.94414 μM	SANGER
NCI-H226	Growth Inhibition Assay			IC50=8.21122 μM	SANGER
MKN45	Growth Inhibition Assay			IC50=8.26602 μM	SANGER
D-392MG	Growth Inhibition Assay			IC50=8.52722 μM	SANGER
RCC10RGB	Growth Inhibition Assay			IC50=8.86695 μM	SANGER
CAL-51	Growth Inhibition Assay			IC50=9.10251 μM	SANGER
COLO-678	Growth Inhibition Assay			IC50=9.32811 μM	SANGER
SK-MEL-24	Growth Inhibition Assay			IC50=9.55856 μM	SANGER
SK-MEL-30	Growth Inhibition Assay			IC50=9.94476 μM	SANGER
MMAC-SF	Growth Inhibition Assay			IC50=10.3961 μM	SANGER
NTERA-S-cl-D1	Growth Inhibition Assay			IC50=10.6508 μM	SANGER
NB12	Growth Inhibition Assay			IC50=11.503 μM	SANGER
UACC-257	Growth Inhibition Assay			IC50=11.8695 μM	SANGER
LAN-6	Growth Inhibition Assay			IC50=11.9928 μM	SANGER
SW1573	Growth Inhibition Assay			IC50=12.3086 μM	SANGER
NMC-G1	Growth Inhibition Assay			IC50=12.4175 μM	SANGER
SHP-77	Growth Inhibition Assay			IC50=12.5574 μM	SANGER
IGROV-1	Growth Inhibition Assay			IC50=12.6573 μM	SANGER
22RV1	Growth Inhibition Assay			IC50=12.8059 μM	SANGER
SK-MEL-3	Growth Inhibition Assay			IC50=13.3973 μM	SANGER
NCI-H1563	Growth Inhibition Assay			IC50=13.4202 μM	SANGER
IGR-1	Growth Inhibition Assay			IC50=14.0347 μM	SANGER
EW-3	Growth Inhibition Assay			IC50=14.1639 μM	SANGER
JEG-3	Growth Inhibition Assay			IC50=14.4953 μM	SANGER
ES3	Growth Inhibition Assay			IC50=14.6562 μM	SANGER
MDA-MB-175-VII	Growth Inhibition Assay			IC50=14.7111 μM	SANGER
P30-OHK	Growth Inhibition Assay			IC50=15.0496 μM	SANGER
GP5d	Growth Inhibition Assay			IC50=15.1466 μM	SANGER
HMV-II	Growth Inhibition Assay			IC50=15.3333 μM	SANGER
COLO-679	Growth Inhibition Assay			IC50=15.501 μM	SANGER
JAR	Growth Inhibition Assay			IC50=15.7205 μM	SANGER
NCI-H1666	Growth Inhibition Assay			IC50=15.9399 μM	SANGER
SW48	Growth Inhibition Assay			IC50=15.9516 μM	SANGER
NCI-H720	Growth Inhibition Assay			IC50=15.9989 μM	SANGER
HT-1197	Growth Inhibition Assay			IC50=16.0352 μM	SANGER
HL-60	Growth Inhibition Assay			IC50=16.0384 μM	SANGER
BEN	Growth Inhibition Assay			IC50=16.5638 μM	SANGER
HAL-01	Growth Inhibition Assay			IC50=16.8909 μM	SANGER
SW900	Growth Inhibition Assay			IC50=16.9132 μM	SANGER
SBC-1	Growth Inhibition Assay			IC50=17.5429 μM	SANGER
SH-4	Growth Inhibition Assay			IC50=17.5841 μM	SANGER
UACC-62	Growth Inhibition Assay			IC50=17.7637 μM	SANGER
BHT-101	Growth Inhibition Assay			IC50=19.8705 μM	SANGER
DB	Growth Inhibition Assay			IC50=21.5649 μM	SANGER
SK-MEL-1	Growth Inhibition Assay			IC50=21.5872 μM	SANGER
NCI-H747	Growth Inhibition Assay			IC50=22.7379 μM	SANGER
SK-LU-1	Growth Inhibition Assay			IC50=23.3524 μM	SANGER
KG-1	Growth Inhibition Assay			IC50=23.4946 μM	SANGER
LXF-289	Growth Inhibition Assay			IC50=23.714 μM	SANGER
HCC1954	Growth Inhibition Assay			IC50=24.8844 μM	SANGER
Ramos-2G6-4C10	Growth Inhibition Assay			IC50=26.4981 μM	SANGER
DBTRG-05MG	Growth Inhibition Assay			IC50=26.6488 μM	SANGER
NCI-H2052	Growth Inhibition Assay			IC50=27.5684 μM	SANGER
RMG-I	Growth Inhibition Assay			IC50=29.4139 μM	SANGER
H9	Growth Inhibition Assay			IC50=31.3008 μM	SANGER
GR-ST	Growth Inhibition Assay			IC50=32.4453 μM	SANGER
Mo-T	Growth Inhibition Assay			IC50=32.5046 μM	SANGER
SW1088	Growth Inhibition Assay			IC50=32.9801 μM	SANGER
LB2518-MEL	Growth Inhibition Assay			IC50=33.0797 μM	SANGER
NCI-H82	Growth Inhibition Assay			IC50=33.1661 μM	SANGER
LAMA-84	Growth Inhibition Assay			IC50=34.6073 μM	SANGER
KYSE-450	Growth Inhibition Assay			IC50=34.6776 μM	SANGER
LU-99A	Growth Inhibition Assay			IC50=35.0007 μM	SANGER
BE-13	Growth Inhibition Assay			IC50=35.6545 μM	SANGER
GAK	Growth Inhibition Assay			IC50=35.695 μM	SANGER
NCI-H1573	Growth Inhibition Assay			IC50=35.8818 μM	SANGER
AsPC-1	Growth Inhibition Assay			IC50=36.1527 μM	SANGER
HDLM-2	Growth Inhibition Assay			IC50=36.316 μM	SANGER
NCI-H441	Growth Inhibition Assay			IC50=37.0691 μM	SANGER
CAL-27	Growth Inhibition Assay			IC50=37.7231 μM	SANGER
OVCAR-3	Growth Inhibition Assay			IC50=39.3181 μM	SANGER
RPMI-8226	Growth Inhibition Assay			IC50=39.598 μM	SANGER
EFO-21	Growth Inhibition Assay			IC50=40.5814 μM	SANGER
SNU-C2B	Growth Inhibition Assay			IC50=41.5084 μM	SANGER
VM-CUB-1	Growth Inhibition Assay			IC50=43.8037 μM	SANGER
NCI-H2087	Growth Inhibition Assay			IC50=45.0233 μM	SANGER
EW-16	Growth Inhibition Assay			IC50=46.3137 μM	SANGER
SK-N-AS	Growth Inhibition Assay			IC50=46.7167 μM	SANGER
COR-L105	Growth Inhibition Assay			IC50=46.8857 μM	SANGER
DEL	Growth Inhibition Assay			IC50=48.0429 μM	SANGER
JVM-2	Growth Inhibition Assay			IC50=48.0558 μM	SANGER
KARPAS-45	Growth Inhibition Assay			IC50=49.4538 μM	SANGER
BL21 (DE3)	Function assay			Binding affinity to human recombinant Mdm2 T101W mutant expressed in Escherichia coli BL21 (DE3) cells by antagonist induced dissociation assay, Kd = 0.5 μM.	18680271
BL21 (DE3)	Function assay			Binding affinity to human recombinant Mdm2 K98W mutant expressed in Escherichia coli BL21 (DE3) cells by antagonist induced dissociation assay, Kd = 0.5 μM.	18680271
BL21 (DE3)	Function assay			Binding affinity to human recombinant Mdm2 F55W mutant expressed in Escherichia coli BL21 (DE3) cells by NMR ligand-protein binary titration, Kd = 0.5 μM.	18680271
SJSA1	Antiproliferative assay		3 days	Antiproliferative activity against human SJSA1 cells after 3 days by EdU incorporation assay in presence of 10% human serum, IC50 = 0.7 μM.	22524527
HCT116	Function assay		24 hrs	Induction of p53-Ser15 phosphorylation in human HCT116 cells after 24 hrs, IC50 = 1 μM.	19856920
BL21 (DE3)	Function assay			Binding affinity to human recombinant Mdm2 T101W mutant expressed in Escherichia coli BL21 (DE3) cells by NMR ligand-protein binary titration, Kd = 1 μM.	18680271
BL21 (DE3)	Function assay			Binding affinity to human recombinant Mdm2 K98W mutant expressed in Escherichia coli BL21 (DE3) cells by NMR ligand-protein binary titration, Kd = 1 μM.	18680271
SJSA1	Cytotoxicity assay			Cytotoxicity against human SJSA1 cells, IC50 = 1 μM.	25396320
RKO	Cytotoxicity assay			Cytotoxicity against human RKO cells, IC50 = 1 μM.	25396320
HCT116	Cytotoxicity assay			Cytotoxicity against human HCT116 cells, IC50 = 1 μM.	25396320
SJSA1	Growth inhibition assay			Growth inhibition of human SJSA1 cells expressing MDM2 by SRB assay, GI50 = 1.3 μM.	21875801
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells expressing wild type p53 incubated for 72 hrs by MTS assay, IC50 = 1.3 μM.	30221935
HCT116	Function assay		8 hrs	Induction of p53 in human HCT116 cells coexpressing pp53TA-luc assessed as inhibition of cell proliferation after 8 hrs by firefly/renilla luciferase reporter assay, IC50 = 1.39 μM.	19856920
HGF	Cytotoxicity assay		24 hrs	Cytotoxicity against human HGF cells after 24 hrs by MTT assay, IC50 = 1.4 μM.	23802716
SJSA1	Antiproliferative assay		72 hrs	Antiproliferative activity against human SJSA1 cells expressing wild type p53 incubated for 72 hrs by MTS assay, IC50 = 1.44 μM.	30221935
SJSA1	Cytotoxicity assay		5 days	Cytotoxicity against human SJSA1 cells expressing wild type p53 assessed as cell viability after 5 days by MTT assay, IC50 = 1.5 μM.	24900694
RKO	Cytotoxicity assay		5 days	Cytotoxicity against human RKO cells expressing wild type p53 assessed as cell viability after 5 days by MTT assay, IC50 = 1.5 μM.	24900694
HCT116	Cytotoxicity assay		5 days	Cytotoxicity against human HCT116 cells expressing wild type p53 assessed as cell viability after 5 days by MTT assay, IC50 = 1.5 μM.	24900694
A2780	Growth inhibition assay			Growth inhibition of human A2780 cells expressing MDM2 by SRB assay, GI50 = 1.6 μM.	21875801
HCT116	Cytotoxicity assay		72 hrs	Cytotoxicity against human HCT116 cells after 72 hrs by CellTiterGlo luciferase-based assay, IC50 = 1.6 μM.	24139845
SJSA1	Antiproliferative assay			Antiproliferative activity against human SJSA1 cells, IC50 = 1.9 μM.	24656661
SJSA1	Antiproliferative assay		72 hrs	Antiproliferative activity against human SJSA1 cells expressing wild type p53 after 72 hrs by CellTitre-Glo assay, IC50 = 1.9 μM.	26985323
HCT116	Growth inhibition assay			Growth inhibition of human HCT116 cells expressing MDM2 by SRB assay, GI50 = 2.1 μM.	21875801
NGP	Cytotoxicity assay		72 hrs	Cytotoxicity against human NGP cells after 72 hrs by sulforhodamine B assay, GI50 = 2.1 μM.	21314128
LS	Cytotoxicity assay		72 hrs	Cytotoxicity against human LS cells after 72 hrs by sulforhodamine B assay, GI50 = 2.3 μM.	21314128
SJSA1	Cytotoxicity assay		72 hrs	Cytotoxicity against human SJSA1 cells after 72 hrs by sulforhodamine B assay, GI50 = 2.6 μM.	21314128
MCF7	Cytotoxicity assay		24 hrs	Cytotoxicity against human MCF7 cells after 24 hrs by MTT assay, IC50 = 2.9 μM.	23802716
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity in p53 (+/+) human HCT116 cells incubated for 72 hrs by MTS assay, IC50 = 4 μM.	28987608
A549	Cytotoxicity assay		72 hrs	Cytotoxicity against human A549 cells expressing wild type p53 assessed as growth inhibition after 72 hrs by SRB assay, IC50 = 4.62 μM.	23601819
A549	Cytotoxicity assay		72 hrs	Cytotoxicity against human A549 cells expressing wild type p53 after 72 hrs by SRB assay, IC50 = 4.62 μM.	23611770
MCF7	Antiproliferative assay		72 hrs	Antiproliferative activity against human MCF7 cells assessed as growth inhibition after 72 hrs by MTT assay, IC50 = 5.01 μM.	27101893
HepG2	Cytotoxicity assay		48 hrs	Cytotoxicity against human HepG2 cells after 48 hrs by MTT assay, IC50 = 5.1 μM.	25479770
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells expressing wild type p53 assessed as growth inhibition after 72 hrs by MTT assay, IC50 = 5.21 μM.	27101893
UKF-NB-3	Cytotoxicity assay		96 hrs	Cytotoxicity against doxorubicin-resistant human UKF-NB-3 cells after 96 hrs by MTT assay, IC50 = 5.56 μM.	20947617
SJSA1	Function assay		7 hrs	Activity at p53 in human SJSA1 cells assessed as induction of p21 mRNA expression after 7 hrs by qRT-PCR analysis in presence of 10% human serum, IC50 = 7.1 μM.	22524527
HCT116	Growth inhibition assay		48 hrs	Growth inhibition of human HCT116 cells harboring p53+/+ after 48 hrs by Hoechst 33258 staining based fluorescence assay, GI50 = 8 μM.	29089230
MCF7	Antiproliferative assay		72 hrs	Antiproliferative activity against human MCF7 cells after 72 hrs by CCK8 assay, IC50 = 8.5 μM.	30045621
HCT116	Antiproliferative assay		24 hrs	Antiproliferative activity against human HCT116 p53+/+ cells assessed as growth inhibition after 24 hrs by EdU/Hoechst 33342 staining-based fluorescence analysis, GI50 = 9 μM.	29691156
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells after 72 hrs by CCK8 assay, IC50 = 9.07 μM.	30045621
A549	Antiproliferative assay		72 hrs	Antiproliferative activity against human A549 cells after 72 hrs by CCK8 assay, IC50 = 9.58 μM.	30045621
HepG2	Cytotoxicity assay		24 hrs	Cytotoxicity against human HepG2 cells after 24 hrs by MTT assay, IC50 = 10.2 μM.	23802716
NCI-H1299	Antiproliferative assay		72 hrs	Antiproliferative activity against p53 -/- human NCI-H1299 cells incubated for 72 hrs by MTS assay, IC50 = 10.4 μM.	30221935
MCF7	Cytotoxicity assay		48 hrs	Cytotoxicity against human MCF7 cells after 48 hrs by MTT assay, IC50 = 11.6 μM.	25618595
Saos2	Antiproliferative assay		72 hrs	Antiproliferative activity against human p53-deficient Saos2 cells after 72 hrs by MTT assay, IC50 = 12.1 μM.	22940704
HCT116	Cytotoxicity assay		72 hrs	Cytotoxicity against human p53 expressing HCT116 cells after 72 hrs by sulforhodamine B assay, GI50 = 13 μM.	21314128
U2OS	Antiproliferative assay		72 hrs	Antiproliferative activity against human U2OS cells expressing wild type p53 after 72 hrs by MTT assay, IC50 = 14.31 μM.	23046248
H1299	Cytotoxicity assay		72 hrs	Cytotoxicity against human H1299 cells after 72 hrs by CellTiterGlo luciferase-based assay, IC50 = 14.7 μM.	24139845
A549	Antiproliferative assay		72 hrs	Antiproliferative activity against human A549 cells expressing wild type p53 after 72 hrs by MTT assay, IC50 = 15 μM.	22940704
A549	Antiproliferative assay		72 hrs	Antiproliferative activity against human A549 cells expressing wild type p53 after 72 hrs by MTT assay, IC50 = 15.12 μM.	23046248
U937	Cytotoxicity assay		24 hrs	Cytotoxicity against human U937 cells after 24 hrs by MTT assay, IC50 = 15.6 μM.	23802716
Saos2	Cytotoxicity assay		72 hrs	Cytotoxicity against human Saos2 cells after 72 hrs by sulforhodamine B assay, GI50 = 18 μM.	21314128
U2OS	Antiproliferative assay		72 hrs	Antiproliferative activity against human U2OS cells expressing wild type p53 after 72 hrs by MTT assay, IC50 = 19.6 μM.	22940704
NCI-H1299	Antiproliferative assay		72 hrs	Antiproliferative activity against human NCI-H1299 cells after 72 hrs by CCK8 assay, IC50 = 19.76 μM.	30045621
PC3	Cytotoxicity assay		72 hrs	Cytotoxicity against human p53-null PC3 cells assessed as growth inhibition after 72 hrs by SRB assay, IC50 = 20.04 μM.	23601819
PC3	Cytotoxicity assay		72 hrs	Cytotoxicity against human p53 deficient PC3 cells after 72 hrs by SRB assay, IC50 = 20.04 μM.	23611770
A2780/CP70	Growth inhibition assay			Growth inhibition of human A2780/CP70 cells expressing isogenically paired p53 mutant by SRB assay, GI50 = 20.4 μM.	21875801
NCI-H1299	Antiproliferative assay		72 hrs	Antiproliferative activity against human p53-deficient NCI-H1299 cells after 72 hrs by MTT assay, IC50 = 20.4 μM.	22940704
NCI-H1299	Antiproliferative assay		72 hrs	Antiproliferative activity against p53 deficient human NCI-H1299 cells after 72 hrs by MTT assay, IC50 = 20.48 μM.	23046248
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human p53-null HCT116 cells assessed as growth inhibition after 72 hrs by MTT assay, IC50 = 21.5 μM.	27101893
SJSA1	Cytotoxicity assay		24 hrs	Cytotoxicity in human SJSA1 cells over expressing human DM2 after 24 hrs by CellTiter-Glo assay, EC50 = 22.8 μM.	29150077
C643	Cytotoxicity assay		24 hrs	Cytotoxicity against human C643 cells after 24 hrs by MTT assay, IC50 = 23 μM.	23802716
MDA-MB-231	Cytotoxicity assay		48 hrs	Cytotoxicity against human MDA-MB-231 cells after 48 hrs by MTT assay, IC50 = 23.5 μM.	25618595
U2OS	Antitumor assay		72 hrs	Antitumor activity against human U2OS cells expressing wild type p53 after 72 hrs by MTT assay, IC50 = 24.61 μM.	21996465
HCT116	Growth inhibition assay			Growth inhibition of p53 deficient human HCT116 cells expressing MDM2 by SRB assay, GI50 = 25.6 μM.	21875801
HCT116	Growth inhibition assay		48 hrs	Growth inhibition of human p53 knockdown HCT116 cells after 48 hrs by Hoechst 33258 staining based fluorescence assay, GI50 = 27 μM.	29089230
Calu	Cytotoxicity assay		24 hrs	Cytotoxicity against human Calu cells after 24 hrs by MTT assay, IC50 = 27.2 μM.	23802716
HCT116	Antiproliferative assay		24 hrs	Antiproliferative activity against human HCT116 p53-/- cells assessed as growth inhibition after 24 hrs by EdU/Hoechst 33342 staining-based fluorescence analysis, GI50 = 29 μM.	29691156
MDA-MB-435S	Antiproliferative assay		72 hrs	Antiproliferative activity against human MDA-MB-435S cells assessed as growth inhibition after 72 hrs by MTT assay, IC50 = 29.6 μM.	27101893
PC3	Cytotoxicity assay		24 hrs	Cytotoxicity against human PC3 cells after 24 hrs by MTT assay, IC50 = 30.3 μM.	23802716
Saos2	Antiproliferative assay		72 hrs	Antiproliferative activity against p53 deficient human Saos2 cells after 72 hrs by MTT assay, IC50 = 31.62 μM.	23046248
HCT116	Antiproliferative assay		48 hrs	Antiproliferative activity against human HCT116 cells expressing wild type p53 after 48 hrs by MTT assay, GI50 = 32.11 μM.	24852275
HCT116	Cytotoxicity assay		72 hrs	Cytotoxicity against p53 deficient human HCT116 cells after 72 hrs by sulforhodamine B assay, GI50 = 34 μM.	21314128
HCT116	Antiproliferative assay		24 hrs	Antiproliferative activity against human HCT116 cells expressing p53 after 24 hrs by MTS assay, GI50 = 39.65 μM.	24268795
A549	Antiproliferative assay		48 hrs	Antiproliferative activity against human A549 cells after 48 hrs by MTT assay, GI50 = 47.59 μM.	24852275
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity in p53 (-/-) human HCT116 cells incubated for 72 hrs by MTS assay, IC50 = 47.8 μM.	28987608
MCF7	Antiproliferative assay		48 hrs	Antiproliferative activity against human MCF7 cells after 48 hrs by MTT assay, GI50 = 48.74 μM.	24852275
SJSA1	Function assay	10 uM		Inhibition of MDM2-p53 interaction in human SJSA1 cells assessed as p53 activation at 10 uM by Western blot	19928922
MCF7	Cell cycle assay	4 uM	14 days	Induction of cell cycle arrest in human MCF7 cells transfected with p53-targeting iRNA-based shRNA at 4 uM after 14 days by BrdU incorporation assay	16474381
MCF7	Cell cycle assay	4 uM	14 days	Induction of cell cycle arrest in human MCF7 cells transfected hnRNPK-targeting iRNA-based shRNA at 4 uM after 14 days by BrdU incorporation assay	16474381
MCF7	Antiproliferative assay	4 uM	14 days	Antiproliferative activity against human MCF7 cells at 4 uM after 14 days by coomassie staining	16474381
MCF7	Antiproliferative assay	4 uM	14 days	Antiproliferative activity against human MCF7 cells assessed as appearance of flattened phenotype at 4 uM after 14 days by coomassie staining	16474381
MCF7	Function assay			Induction of p53 expression in human MCF7 cells transfected with 53BP1-targeting iRNA-based shRNA by Western blot analysis	16474381
MCF7	Function assay			Induction of p53 expression in human MCF7 cells by Western blot analysis	16474381
MCF7	Function assay		18 hrs	Induction of p53 accumulation in human MCF7 cells after 18 hrs by Western blot analysis in presence of 2.5 mM caffeine	16474381
BJ	Function assay	3 uM		Induction of p21CIP1 expression in irradiated human BJ cells at 3 uM by Western blot analysis	16474381
MCF7	Cell cycle assay	4 uM	14 days	Induction of cell cycle arrest in human MCF7 cells transfected with 53BP1-targeting iRNA-based shRNA at 4 uM after 14 days by BrdU incorporation assay	16474381
BJ	Function assay	3 uM		Induction of p21 expression in irradiated human BJ cells at 3 uM by Western blot analysis	16474381
SNJSA1	Function assay	1 to 10 uM	6 hrs	Inhibition of Mdm2 -p53 protein interaction in human SNJSA1 cells assessed as induction of p21 protein at 1 to 10 uM after 6 hrs by Western blot analysis	21314128
SNJSA1	Function assay	1 to 10 uM	6 hrs	Inhibition of Mdm2 -p53 protein interaction in human SNJSA1 cells assessed as induction of p53 protein at 1 to 10 uM after 6 hrs by Western blot analysis	21314128
SNJSA1	Function assay	1 to 10 uM	6 hrs	Inhibition of Mdm2 -p53 protein interaction in human SNJSA1 cells assessed as induction of MDM2 protein at 1 to 10 uM after 6 hrs by Western blot analysis	21314128
MCF7	Function assay		24 hrs	Upregulation of p53 in human MCF7 cells after 24 hrs by Western blot analysis	27010502
MCF7	Function assay		24 hrs	Upregulation of Mdm2 in human MCF7 cells after 24 hrs by Western blot analysis	27010502
MCF7	Function assay		24 hrs	Increase in p53 expression in human MCF7 cells nuclei after 24 hrs by DAPI staining based immunofluorescence microscopy	27010502
MCF7	Function assay		24 hrs	Upregulation of p21 in human MCF7 cells after 24 hrs by Western blot analysis	27010502
HCT116	Function assay	50 uM	24 hrs	Inhibition of p53-MDM2 (unknown origin) interaction expressed in p53-deficient human HCT116 cells at 50 uM after 24 hrs by BiFC-based FACS flow cytometric analysis	24268795
U2OS	Function assay	2.5 uM	12 hrs	Stabilization of p53 in human U2OS cells assessed as intracellular p53 level at 2.5 uM after 12 hrs by Western blotting	26288684
HCT116	Function assay	10 uM	20 hrs	Inhibition of V1-p53/MDM2-V2 (unknown origin) interaction expressed in human HCT116 p53-/- cells at 10 uM after 20 hrs by biomolecular fluorescence complementation-based flow cytometric analysis	28800455
IMR32	Function assay		24 hrs	Increase in p53 expression in human IMR32 cells nuclei after 24 hrs by DAPI staining based immunofluorescence microscopy	27010502
MCF7	Function assay	10 uM	16 hrs	Inhibition of MDM2-p53 interaction in human MCF7 cells assessed as reversal of MDM2-dependent inhibition of p53-induced transcriptional activity at 10 uM after 16 hrs by dual-luciferase reporter gene assay	23540934
MCF7	Apoptosis assay	3 uM	24 to 72 hrs	Induction of apoptosis in human MCF7 cells assessed as reduction in Bcl-xL expression level at 3 uM after 24 to 72 hrs by Western blotting analysis	23802716
MCF7	Apoptosis assay	3 uM	24 to 72 hrs	Induction of apoptosis in human MCF7 cells assessed as early apoptotic cells at 3 uM after 24 to 72 hrs by annexin V-FITC/propidium iodide staining based FACS flow cytometry (Rvb = 4 %)	23802716
MCF7	Apoptosis assay	3 uM	24 to 72 hrs	Induction of apoptosis in human MCF7 cells assessed as increase in p21 expression level at 3 uM after 24 to 72 hrs by Western blotting analysis	23802716
MCF7	Apoptosis assay	3 uM	24 to 72 hrs	Induction of apoptosis in human MCF7 cells assessed as increase in Bcl-xS expression level at 3 uM after 24 to 72 hrs by Western blotting analysis	23802716
MCF7	Apoptosis assay	50 nM	24 to 72 hrs	Induction of apoptosis in human MCF7 cells assessed as accumulation of caspase-3 cleavage product at 50 nM after 24 to 72 hrs by Western blotting analysis	23802716
MCF7	Cell cycle assay	3 uM	24 to 72 hrs	Cell cycle arrest in human MCF7 cells assessed as accumulation at G0/G1 phase at 3 uM after 24 to 72 hrs by propidium iodide staining-based FACS analysis	23802716
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 100 mg/mL (171.97 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 100 mg/mL

Water : Insoluble

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O Validated by Selleck labs. Should you need adjustments to this formulation, contact our sales team for custom testing.	10.000mg/ml (17.20mM)	Taking the 1 mL working solution as an example, add 50 μL of 200 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

Dosage: mg/kg

Average weight of animals: g

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Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO

% Tween 80

% ddH₂O

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Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH₂O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
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Chemical Information, Storage & Stability

Molecular Weight	581.49	Formula	C₃₀H₃₀Cl₂N₄O₄	Storage (From the date of receipt)	3 years-20°Cpowder
CAS No.	675576-98-4	--		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	(-)-Nutlin-3			Smiles	CC(C)OC1=C(C=CC(=C1)OC)C2=NC(C(N2C(=O)N3CCNC(=O)C3)C4=CC=C(C=C4)Cl)C5=CC=C(C=C5)Cl

Mechanism of Action

Features	Highly selective MDM2 inhibitor with a much lower effect on MDMX. Most effective on tumors with wild type p53.
Targets/IC₅₀/Ki	p53-MDM2 interaction (Cell-free assay) 90 nM
In vitro	Nutlin-3a displaces p53 from the binding pocket of MDM2 and thereby releases p53 from inhibition and proteasomal degradation, leading to induction of its downstream targets, cell cycle arrest, and apoptosis. Seven days of incubation with 10 μM of this compound led to >90% inhibition of NIH3T3 cells’ growth. This compound stabilizes and activates p53, and induces p21 expression in a dose-dependent manner. It effectively depletes the S-phase compartment to 0.2-2% and increases the G1- and G2/M-phase compartments. This chemical induces apoptosis in ~60% of SJSA-1 and MHM cells after 40 h, which increased further after 60 h (85% and 65%, respectively) .
Kinase Assay	Biacore studies
Kinase Assay	Competition assays are performed on a Biacore S51. A Series S Sensor chip CM5 is derivatized for immobilization of a PentaHis antibody for capture of the His-tagged p53. The level of capture is ~ 200 response units (1 response unit corresponds to 1 pg of protein per mm ²). The concentration of MDM2 protein is kept constant at 300 nM. Test compounds are dissolved in DMSO at 10 mM and further diluted to make a concentration series of inhibitor in each MDM2 test sample. The assays are run at 25 °C in running buffer (10 mM Hepes, 0.15 M NaCl, 2% DMSO). MDM2-p53 binding in the presence of this compound is calculated as a percentage of binding in the absence of this compound and IC50 is calculated using Microsoft Excel
In vivo	Nutlin-3a suppresses xenograft growth in a dose-dependent fashion with the highest dose (200 mg/kg) showing a substantial tumor shrinkage . This compound is a selective activator of the p53 pathway in vivo and highly efficacious against SJSA-1 osteosarcoma tumors. Tumors with wild-type p53 and mdm2 gene amplification will respond best to therapy with this chemical.
References	[1] https://pubmed.ncbi.nlm.nih.gov/16443686/ [2] https://pubmed.ncbi.nlm.nih.gov/21624110/ [3] https://pubmed.ncbi.nlm.nih.gov/14704432/

Applications

Methods	Biomarkers	PMID
Western blot	p53 / MDM2 / p21 / Bax / Bak / Noxa / cleaved caspase / pro-caspase BMI-1 p53-Ser15 / p53-Ser37 / p53-Lys382 / MDM2 / CDC2 / PLK1 / HSC70	16014563
Immunofluorescence	p-p21 / β-actin P53 Ac-p300	31083332
Growth inhibition assay	Cell viability	26248031