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PLX-4720 B-Raf Inhibitor

Name: PLX-4720
Brand: Selleck Chemicals
SKU: S1152
Rating: 5 (198 reviews)

N. Cat.: S1152

PLX4720 is a potent and selective inhibitor of B-Raf^V600E with IC50 of 13 nM in a cell-free assay, equally potent to c-Raf-1(Y340D and Y341D mutations), 10-fold selectivity for B-RafV600E than wild-type B-Raf.

PLX-4720 Raf inhibitor Chemical Structure

Struttura chimica

Peso molecolare: 413.83

Vai a

Controllo Qualità (Quality Control)

Lotto: Purezza: 99.98%

99.98

Prodotti spesso usati con PLX-4720

Dactolisib (BEZ235)

Treatment with this compound and Dactolisib decreases both pERK and pAKT levels in Colo-205 xenografts.

Navitoclax (ABT-263)

It and Navitoclax can enhance the clinical efficacy of BRAF inhibitors.

NMS-E973

It and NMS-E973 show a synergic antiproliferative activity in A375 melanoma cells.

SB431542

Cotreatment with this compound and SB431542 abolish the increase in cell growth caused by low-dose PLX-4720 in Colo829 and A375(M2) cells.

Target correlati	ERK p38 MAPK JNK MEK Ras KRas S6 Kinase MAP4K TAK1 Mixed Lineage Kinase
Altro Raf Inibitori	LY3009120 Exarafenib (KIN-2787) GDC-0879 Avutometinib (Ro5126766, CH5126766) AZ 628 SB590885 TAK-632 GW5074 RAF265 (CHIR-265) PLX7904

Coltura cellulare, trattamento e concentrazione di lavoro
(Cell Culture, Treatment & Working Concentration)

Linee cellulari	Tipo di saggio	Tempo di incubazione	Descrizione dell'attività	PMID
DU-4475	Growth Inhibition Assay		IC50=0.07457 μM	SANGER
EoL-1-cell	Growth Inhibition Assay		IC50=0.14166 μM	SANGER
C32	Growth Inhibition Assay		IC50=0.15131 μM	SANGER
M14	Growth Inhibition Assay		IC50=0.21757 μM	SANGER
CP50-MEL-B	Growth Inhibition Assay		IC50=0.29784 μM	SANGER
A101D	Growth Inhibition Assay		IC50=0.32589 μM	SANGER
G-361	Growth Inhibition Assay		IC50=0.34637 μM	SANGER
HT-144	Growth Inhibition Assay		IC50=0.36329 μM	SANGER
ACN	Growth Inhibition Assay		IC50=0.38477 μM	SANGER
COLO-829	Growth Inhibition Assay		IC50=0.38968 μM	SANGER
MEL-HO	Growth Inhibition Assay		IC50=0.41179 μM	SANGER
SH-4	Growth Inhibition Assay		IC50=0.41422 μM	SANGER
SK-MEL-3	Growth Inhibition Assay		IC50=0.51568 μM	SANGER
A375	Growth Inhibition Assay		IC50=0.67359 μM	SANGER
MMAC-SF	Growth Inhibition Assay		IC50=0.68614 μM	SANGER
BHT-101	Growth Inhibition Assay		IC50=0.70702 μM	SANGER
K5	Growth Inhibition Assay		IC50=0.76148 μM	SANGER
BV-173	Growth Inhibition Assay		IC50=0.79644 μM	SANGER
RVH-421	Growth Inhibition Assay		IC50=0.86796 μM	SANGER
HCC2218	Growth Inhibition Assay		IC50=0.87844 μM	SANGER
WM-115	Growth Inhibition Assay		IC50=0.88692 μM	SANGER
SK-MEL-28	Growth Inhibition Assay		IC50=1.04569 μM	SANGER
COLO-679	Growth Inhibition Assay		IC50=1.10464 μM	SANGER
MZ7-mel	Growth Inhibition Assay		IC50=1.14963 μM	SANGER
SK-MEL-30	Growth Inhibition Assay		IC50=1.33386 μM	SANGER
NCI-H209	Growth Inhibition Assay		IC50=1.6086 μM	SANGER
HTC-C3	Growth Inhibition Assay		IC50=1.66294 μM	SANGER
KARPAS-45	Growth Inhibition Assay		IC50=2.04978 μM	SANGER
NCI-SNU-5	Growth Inhibition Assay		IC50=2.11969 μM	SANGER
KP-4	Growth Inhibition Assay		IC50=2.30787 μM	SANGER
PA-1	Growth Inhibition Assay		IC50=2.72673 μM	SANGER
HuO-3N1	Growth Inhibition Assay		IC50=2.87946 μM	SANGER
NCI-H358	Growth Inhibition Assay		IC50=2.92232 μM	SANGER
CTB-1	Growth Inhibition Assay		IC50=3.40176 μM	SANGER
697	Growth Inhibition Assay		IC50=3.55266 μM	SANGER
CP66-MEL	Growth Inhibition Assay		IC50=4.15927 μM	SANGER
NB13	Growth Inhibition Assay		IC50=4.49179 μM	SANGER
DBTRG-05MG	Growth Inhibition Assay		IC50=4.53325 μM	SANGER
A2058	Growth Inhibition Assay		IC50=4.72164 μM	SANGER
KG-1	Growth Inhibition Assay		IC50=4.73908 μM	SANGER
8305C	Growth Inhibition Assay		IC50=5.1873 μM	SANGER
RPMI-7951	Growth Inhibition Assay		IC50=5.80283 μM	SANGER
CHL-1	Growth Inhibition Assay		IC50=5.97603 μM	SANGER
TI-73	Growth Inhibition Assay		IC50=6.00902 μM	SANGER
HT-1080	Growth Inhibition Assay		IC50=6.10946 μM	SANGER
ES5	Growth Inhibition Assay		IC50=6.14924 μM	SANGER
8-MG-BA	Growth Inhibition Assay		IC50=6.18129 μM	SANGER
NB7	Growth Inhibition Assay		IC50=6.21373 μM	SANGER
H4	Growth Inhibition Assay		IC50=6.22493 μM	SANGER
CAL-72	Growth Inhibition Assay		IC50=6.45423 μM	SANGER
HCC1806	Growth Inhibition Assay		IC50=6.81931 μM	SANGER
BCPAP	Growth Inhibition Assay		IC50=7.21764 μM	SANGER
LB2241-RCC	Growth Inhibition Assay		IC50=7.36907 μM	SANGER
COLO-741	Growth Inhibition Assay		IC50=8.01679 μM	SANGER
HSC-3	Growth Inhibition Assay		IC50=8.07068 μM	SANGER
SW982	Growth Inhibition Assay		IC50=8.41516 μM	SANGER
GCT	Growth Inhibition Assay		IC50=8.75314 μM	SANGER
KY821	Growth Inhibition Assay		IC50=9.05178 μM	SANGER
JVM-3	Growth Inhibition Assay		IC50=9.56999 μM	SANGER
RS4-11	Growth Inhibition Assay		IC50=9.6048 μM	SANGER
VA-ES-BJ	Growth Inhibition Assay		IC50=10.0149 μM	SANGER
A431	Growth Inhibition Assay		IC50=10.4212 μM	SANGER
LXF-289	Growth Inhibition Assay		IC50=10.458 μM	SANGER
SK-MEL-24	Growth Inhibition Assay		IC50=10.8274 μM	SANGER
NOS-1	Growth Inhibition Assay		IC50=10.8472 μM	SANGER
KNS-62	Growth Inhibition Assay		IC50=11.2404 μM	SANGER
SK-HEP-1	Growth Inhibition Assay		IC50=11.3527 μM	SANGER
A3-KAW	Growth Inhibition Assay		IC50=11.7178 μM	SANGER
SK-LU-1	Growth Inhibition Assay		IC50=12.2655 μM	SANGER
TYK-nu	Growth Inhibition Assay		IC50=12.3932 μM	SANGER
NMC-G1	Growth Inhibition Assay		IC50=12.6062 μM	SANGER
BB65-RCC	Growth Inhibition Assay		IC50=12.7169 μM	SANGER
QIMR-WIL	Growth Inhibition Assay		IC50=12.8833 μM	SANGER
D-566MG	Growth Inhibition Assay		IC50=13.9576 μM	SANGER
KYSE-140	Growth Inhibition Assay		IC50=14.0753 μM	SANGER
SCC-4	Growth Inhibition Assay		IC50=14.3359 μM	SANGER
U251	Growth Inhibition Assay		IC50=14.8492 μM	SANGER
D-542MG	Growth Inhibition Assay		IC50=14.9222 μM	SANGER
LAMA-84	Growth Inhibition Assay		IC50=14.9932 μM	SANGER
NCI-H720	Growth Inhibition Assay		IC50=15.2684 μM	SANGER
DEL	Growth Inhibition Assay		IC50=15.4293 μM	SANGER
SBC-1	Growth Inhibition Assay		IC50=15.4305 μM	SANGER
ECC10	Growth Inhibition Assay		IC50=15.4458 μM	SANGER
Daoy	Growth Inhibition Assay		IC50=15.7616 μM	SANGER
SCH	Growth Inhibition Assay		IC50=15.7835 μM	SANGER
MZ2-MEL	Growth Inhibition Assay		IC50=16.0646 μM	SANGER
CAL-12T	Growth Inhibition Assay		IC50=16.4862 μM	SANGER
KE-37	Growth Inhibition Assay		IC50=16.8107 μM	SANGER
LS-411N	Growth Inhibition Assay		IC50=17.118 μM	SANGER
NCI-H2228	Growth Inhibition Assay		IC50=17.3071 μM	SANGER
SK-MEL-2	Growth Inhibition Assay		IC50=17.4965 μM	SANGER
HN	Growth Inhibition Assay		IC50=17.7248 μM	SANGER
NCI-H1648	Growth Inhibition Assay		IC50=17.818 μM	SANGER
IA-LM	Growth Inhibition Assay		IC50=18.3172 μM	SANGER
EW-13	Growth Inhibition Assay		IC50=18.5708 μM	SANGER
YKG-1	Growth Inhibition Assay		IC50=19.5711 μM	SANGER
KNS-81-FD	Growth Inhibition Assay		IC50=19.5858 μM	SANGER
23132-87	Growth Inhibition Assay		IC50=19.7642 μM	SANGER
NUGC-3	Growth Inhibition Assay		IC50=19.9887 μM	SANGER
5637	Growth Inhibition Assay		IC50=20.0478 μM	SANGER
NCI-H1755	Growth Inhibition Assay		IC50=20.4764 μM	SANGER
RH-18	Growth Inhibition Assay		IC50=20.5748 μM	SANGER
RXF393	Growth Inhibition Assay		IC50=20.6756 μM	SANGER
LU-134-A	Growth Inhibition Assay		IC50=20.7056 μM	SANGER
TE-12	Growth Inhibition Assay		IC50=20.7201 μM	SANGER
MOLT-4	Growth Inhibition Assay		IC50=21.1915 μM	SANGER
IGR-1	Growth Inhibition Assay		IC50=21.3796 μM	SANGER
HOP-92	Growth Inhibition Assay		IC50=21.4987 μM	SANGER
SK-MES-1	Growth Inhibition Assay		IC50=21.7381 μM	SANGER
LU-65	Growth Inhibition Assay		IC50=21.8624 μM	SANGER
MS-1	Growth Inhibition Assay		IC50=22.1203 μM	SANGER
LoVo	Growth Inhibition Assay		IC50=22.244 μM	SANGER
A704	Growth Inhibition Assay		IC50=22.5155 μM	SANGER
HT-1376	Growth Inhibition Assay		IC50=22.6059 μM	SANGER
IST-MEL1	Growth Inhibition Assay		IC50=22.6751 μM	SANGER
Ramos-2G6-4C10	Growth Inhibition Assay		IC50=22.7366 μM	SANGER
T47D	Growth Inhibition Assay		IC50=22.7979 μM	SANGER
HT-1197	Growth Inhibition Assay		IC50=23.0817 μM	SANGER
LB2518-MEL	Growth Inhibition Assay		IC50=23.6412 μM	SANGER
J-RT3-T3-5	Growth Inhibition Assay		IC50=24.7595 μM	SANGER
SK-NEP-1	Growth Inhibition Assay		IC50=24.8744 μM	SANGER
NCI-H526	Growth Inhibition Assay		IC50=25.0023 μM	SANGER
IST-SL1	Growth Inhibition Assay		IC50=25.2751 μM	SANGER
HH	Growth Inhibition Assay		IC50=25.3192 μM	SANGER
NCI-H82	Growth Inhibition Assay		IC50=25.938 μM	SANGER
SNU-449	Growth Inhibition Assay		IC50=27.2018 μM	SANGER
COR-L23	Growth Inhibition Assay		IC50=27.2813 μM	SANGER
LOXIMVI	Growth Inhibition Assay		IC50=27.368 μM	SANGER
GR-ST	Growth Inhibition Assay		IC50=27.6706 μM	SANGER
NCI-SNU-1	Growth Inhibition Assay		IC50=27.944 μM	SANGER
ALL-PO	Growth Inhibition Assay		IC50=28.1604 μM	SANGER
ML-2	Growth Inhibition Assay		IC50=28.2814 μM	SANGER
HOP-62	Growth Inhibition Assay		IC50=28.713 μM	SANGER
EGI-1	Growth Inhibition Assay		IC50=28.8845 μM	SANGER
TCCSUP	Growth Inhibition Assay		IC50=28.9272 μM	SANGER
LB996-RCC	Growth Inhibition Assay		IC50=29.5682 μM	SANGER
LCLC-97TM1	Growth Inhibition Assay		IC50=32.1964 μM	SANGER
NCI-H1304	Growth Inhibition Assay		IC50=32.3301 μM	SANGER
KP-N-YS	Growth Inhibition Assay		IC50=32.5973 μM	SANGER
NCI-H1770	Growth Inhibition Assay		IC50=33.1648 μM	SANGER
EM-2	Growth Inhibition Assay		IC50=33.6504 μM	SANGER
ChaGo-K-1	Growth Inhibition Assay		IC50=33.7236 μM	SANGER
ACHN	Growth Inhibition Assay		IC50=33.8385 μM	SANGER
MN-60	Growth Inhibition Assay		IC50=33.8544 μM	SANGER
EW-18	Growth Inhibition Assay		IC50=33.8971 μM	SANGER
KGN	Growth Inhibition Assay		IC50=35.7292 μM	SANGER
U031	Growth Inhibition Assay		IC50=35.8132 μM	SANGER
HMV-II	Growth Inhibition Assay		IC50=36.0774 μM	SANGER
L-363	Growth Inhibition Assay		IC50=37.6455 μM	SANGER
NCI-H1155	Growth Inhibition Assay		IC50=38.0015 μM	SANGER
NCI-H1793	Growth Inhibition Assay		IC50=38.1026 μM	SANGER
P30-OHK	Growth Inhibition Assay		IC50=38.1332 μM	SANGER
AN3-CA	Growth Inhibition Assay		IC50=38.1615 μM	SANGER
UACC-257	Growth Inhibition Assay		IC50=38.79 μM	SANGER
MCF7	Growth Inhibition Assay		IC50=39.8629 μM	SANGER
KP-N-YN	Growth Inhibition Assay		IC50=40.4285 μM	SANGER
T98G	Growth Inhibition Assay		IC50=40.4957 μM	SANGER
HGC-27	Growth Inhibition Assay		IC50=43.274 μM	SANGER
NCI-H1092	Growth Inhibition Assay		IC50=43.2895 μM	SANGER
KARPAS-299	Growth Inhibition Assay		IC50=43.3071 μM	SANGER
LB1047-RCC	Growth Inhibition Assay		IC50=44.9959 μM	SANGER
786-0	Growth Inhibition Assay		IC50=45.65 μM	SANGER
HCC2157	Growth Inhibition Assay		IC50=46.0359 μM	SANGER
NY	Growth Inhibition Assay		IC50=46.1778 μM	SANGER
EFM-19	Growth Inhibition Assay		IC50=46.7533 μM	SANGER
EW-16	Growth Inhibition Assay		IC50=46.7806 μM	SANGER
UM-UC-3	Growth Inhibition Assay		IC50=46.8059 μM	SANGER
HT-29	Growth Inhibition Assay		IC50=47.8792 μM	SANGER
LN-405	Growth Inhibition Assay		IC50=48.0827 μM	SANGER
NCI-H727	Growth Inhibition Assay		IC50=48.7726 μM	SANGER
D-502MG	Growth Inhibition Assay		IC50=48.9676 μM	SANGER
GMS-10	Growth Inhibition Assay		IC50=49.2974 μM	SANGER
MEL-JUSO	Growth Inhibition Assay		IC50=49.347 μM	SANGER
insect cells	Function assay		Inhibition of N-terminal His-tagged BRAF V600E mutant (unknown origin) expressed in baculovirus infected insect cells co-expressing CDC37 using biotinylated-MEK as substrate by AlphaScreen assay, IC50 = 0.013 μM.	29461827
A375	Function assay		Inhibition of BRAF V600E mutant in human A375 cells assessed as reduction in ERK phosphorylation by AlphaScreen assay, IC50 = 0.044 μM.	29461827
A375	Function assay		Inhibition of b-Raf in human A375 cells assessed phosphorylation of ERK, IC50 = 0.046 μM.	22808911
A375	Antiproliferative assay		Antiproliferative activity against human A375 cells expressing B-Raf V600E mutant and wild type Ras, IC50 = 0.5 μM.	22808911
A375	Cytotoxicity assay	72 hrs	Cytotoxicity against human A375 cells harboring BRAF V600E mutant after 72 hrs by CellTiter-Glo assay, IC50 = 0.5 μM.	29461827
HCT116	Antiproliferative assay		Antiproliferative activity against human HCT116 cells expressing wild type b-Raf and KRAS mutant, IC50 = 27 μM.	22808911
A673	qHTS assay		qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
Saos-2	qHTS assay		qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells	29435139
RD	qHTS assay		qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells	29435139
SK-N-SH	qHTS assay		qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells	29435139
NB1643	qHTS assay		qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells	29435139
OHS-50	qHTS assay		qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells	29435139
SJ-GBM2	qHTS assay		qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells	29435139
SK-N-MC	qHTS assay		qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
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Informazioni chimiche, conservazione e stabilità (Chemical Information, Storage & Stability)

Peso molecolare	413.83	Formula	C₁₇H₁₄ClF₂N₃O₃S	Conservazione (Dalla data di ricezione)	3 anni-20°Cpolvere
N. CAS	918505-84-7	Scarica SDF		Conservazione delle soluzioni stock	1 anno-80°Cin solvente 1 mese-20°Cin solvente
Sinonimi	N/A			Smiles	CCCS(=O)(=O)NC1=C(C(=C(C=C1)F)C(=O)C2=CNC3=C2C=C(C=N3)Cl)F

Solubilità (Solubility)

In vitro
Lotto:

DMSO : 83 mg/mL (200.56 mM)
(Il DMSO contaminato da umidità può ridurre la solubilità. Utilizzare DMSO fresco e anidro.)

Water : Insoluble

Ethanol : Insoluble

Calcolatore di Molarità

Massa	Concentrazione	Volume	Peso molecolare
=	×	×

Calcolatore di Diluizione Calcolatore del Peso Molecolare

In vivo Lotto:
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.

Calcolatore di formulazione in vivo (Soluzione chiara)

Passo 1: Inserire le informazioni di seguito (Consigliato: Un animale aggiuntivo per tenere conto della perdita durante l'esperimento)

Dosaggio: mg/kg Peso medio degli animali: g Volume di dosaggio per animale:μL Numero di animali:

Passo 2: Inserire la formulazione in vivo (Questo è solo il calcolatore, non la formulazione. Contattateci prima se non c'è una formulazione in vivo nella sezione Solubilità.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Risultati del calcolo:

Concentrazione di lavoro: mg/ml;

Metodo per preparare il liquido master di DMSO: mg farmaco predissolto in μL DMSO ( Concentrazione del liquido master mg/mL, Vi preghiamo di contattarci prima se la concentrazione supera la solubilità del DMSO del lotto del farmaco. )

Metodo per preparare la formulazione in vivo: Prendere μL DMSO liquido master, quindi aggiungereμL PEG300, mescolare e chiarire, quindi aggiungereμL Tween 80, mescolare e chiarire, quindi aggiungere μL ddH₂O, mescolare e chiarire.

Metodo per preparare la formulazione in vivo: Prendere μL DMSO liquido master, quindi aggiungere μL Olio di mais, mescolare e chiarire.

Nota: 1. Si prega di assicurarsi che il liquido sia limpido prima di aggiungere il solvente successivo.
2. Assicurarsi di aggiungere il/i solvente/i in ordine. È necessario assicurarsi che la soluzione ottenuta, nell'aggiunta precedente, sia una soluzione limpida prima di procedere all'aggiunta del solvente successivo. Metodi fisici come il vortex, gli ultrasuoni o il bagno d'acqua calda possono essere utilizzati per facilitare la dissoluzione.

Meccanismo d'azione (Mechanism of Action)

Targets/IC₅₀/Ki	C-Raf-1 (Y340D/Y341D) (Cell-free assay) 6.7 nM B-Raf (V600E) (Cell-free assay) 13 nM BRK (Cell-free assay) 130 nM B-Raf (Cell-free assay) 160 nM
In vitro	PLX-4720 mostra una selettività >10 volte superiore contro il B-Raf wild type e una selettività >100 volte superiore rispetto ad altre chinasi come Frk, Src, Fak, FGFR e Aurora A con IC50 di 1,3-3,4 μM. Questo composto inibisce significativamente la fosforilazione di ERK in linee cellulari che esprimono B-Raf^V600E con IC50 di 14-46 nM, ma non nelle cellule con B-Raf wild-type. Inibisce significativamente la crescita di linee cellulari tumorali che esprimono l'oncogene B-Raf^V600E, come COLO205, A375, WM2664 e COLO829 con GI50 di 0,31 μM, 0,50 μM, 1,5 μM e 1,7 μM, rispettivamente. Inoltre, questo trattamento chimico a 1 μM induce l'arresto del ciclo cellulare e l'apoptosi esclusivamente nelle cellule 1205Lu positive per B-Raf^V600E, ma non nelle cellule C8161 wild-type per B-Raf. Questo trattamento con il composto (10 μM) induce significativamente un'espressione di BIM >14 volte nelle cellule PTEN⁺, rispetto alle linee cellulari PTEN- (4 volte), fornendo una spiegazione della resistenza delle cellule PTEN^- a questa apoptosi indotta da sostanze chimiche.
Saggio chinasico	Attività chinasiche di Raf in vitro
Saggio chinasico	Le attività chinasiche in vitro del Raf wild type e dei mutanti sono determinate misurando la fosforilazione della proteina MEK biotinilata utilizzando la tecnologia AlphaScreen di Perkin-Elmer. Per ogni enzima (0,1 ng), vengono eseguite reazioni di 20 μL in 20 mM Hepes (pH 7,0), 10 mM MgCl₂, 1 mM DTT, 0,01% Tween-20, 100 nM di proteina biotin-MEK, varie concentrazioni di ATP e concentrazioni crescenti di PLX-4720 a temperatura ambiente. Le reazioni vengono interrotte a 2, 5, 8, 10, 20 e 30 minuti con 5 μL di una soluzione contenente 20 mM Hepes (pH 7,0), 200 mM NaCl, 80 mM EDTA e 0,3% BSA. La soluzione di interruzione include anche l'anticorpo phospho-MEK, perline donatrici rivestite di streptavidina e perline accettrici di proteina A dal kit di rilevazione di proteina A AlphaScreen. L'anticorpo e le perline vengono pre-incubati nella soluzione di interruzione al buio a temperatura ambiente per 30 minuti. La diluizione finale dell'anticorpo è 1/2.000 e la concentrazione finale di ogni perlina è 10 μg/mL. Le piastre del saggio vengono incubate a temperatura ambiente per un'ora e quindi lette su un lettore AlphaQuest di PerkinElmer.
In vivo	La somministrazione orale di PLX-4720 a 20 mg/kg/giorno induce significativi ritardi e regressioni della crescita tumorale in xenotrapianti tumorali COLO205 dipendenti da B-Raf^V600E, senza evidenti effetti avversi nei topi anche a una dose di 1 g/kg. Questo composto a 100 mg/kg due volte al giorno elimina quasi completamente gli xenotrapianti 1205Lu che esprimono B-Raf^V600E, mentre non ha attività contro gli xenotrapianti C8161 che esprimono B-Raf wild-type. Gli effetti antitumorali di questo composto correlano con il blocco della via MAPK in quelle cellule che ospitano la mutazione V600E. Questo trattamento chimico a 30 mg/kg/giorno inibisce significativamente la crescita tumorale degli xenotrapianti 8505c di >90% e diminuisce drasticamente le metastasi polmonari distanti.
Riferimenti	[1] https://pubmed.ncbi.nlm.nih.gov/18287029/ [2] https://pubmed.ncbi.nlm.nih.gov/21317224/ [3] https://pubmed.ncbi.nlm.nih.gov/20498063/

Applicazioni (Applications)

Metodi	Biomarcatori	PMID
Western blot	p-MEK / MEK / p-ERK / ERK / p-FAK(S910) p-EGFR 1173 / EGFR / p-Akt / Akt p27 / Cyclin D1 / pRb pAkt(Ser473) / pAkt(Thr308)	23076151
Immunofluorescence	LAMP1 ZKSCAN3 / TFEB	30979895
Growth inhibition assay	Cell viability	27848137
ELISA	mIFN-γ	23204132

Domande Frequenti (Frequently Asked Questions)

Domanda 1:
What would you recommend to make working solution for intraperitoneal injection into mice?

Risposta:
Due to its very limited solubility in aqueous solution, this compound can be administered as a not fully dissolved suspension via oral gavage. For this reason, we recommend oral gavage for its administration.

C1=C0/X	C1:	LOG(C1):
C2=C1/X	C2:	LOG(C2):
C3=C2/X	C3:	LOG(C3):
C4=C3/X	C4:	LOG(C4):
C5=C4/X	C5:	LOG(C5):
C6=C5/X	C6:	LOG(C6):
C7=C6/X	C7:	LOG(C7):
C8=C7/X	C8:	LOG(C8):