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PLX-4720 B-Raf-remmer

Name: PLX-4720
Brand: Selleck Chemicals
SKU: S1152
Rating: 5 (197 reviews)

Cat.Nr.S1152

PLX4720 is een krachtige en selectieve remmer van B-Raf^V600E met een IC50 van 13 nM in een celvrije test, even krachtig als c-Raf-1 (Y340D- en Y341D-mutaties), 10-voudige selectiviteit voor B-RafV600E dan wild-type B-Raf.

Chemische structuur

Moleculair gewicht: 413.83

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Kwaliteitscontrole

Batch: Zuiverheid: 99.98%

99.98

Producten Vaak Samen Gebruikt Met 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.

Gerelateerde doelwitten	ERK p38 MAPK JNK MEK Ras KRas S6 Kinase MAP4K TAK1 Mixed Lineage Kinase
Overige Raf Inhibitoren	LY3009120 Exarafenib (KIN-2787) GDC-0879 Avutometinib (Ro5126766, CH5126766) AZ 628 SB590885 TAK-632 GW5074 RAF265 (CHIR-265) PLX8394 (Plixorafenib, FORE8394)

Celkweek, behandeling & werkconcentratie

Cellijnen	Assaytype	Incubatietijd	Activiteitsbeschrijving	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
Klik om meer experimentele gegevens over de cellijn te bekijken

Chemische informatie, Opslag en Stabiliteit

Moleculair gewicht	413.83	Formule	C₁₇H₁₄ClF₂N₃O₃S	Opslag (Vanaf de ontvangstdatum)	3 jaar-20°Cpoeder
CAS-nr.	918505-84-7	SDF downloaden		Opslag van stamoplossingen	1 jaar-80°Cin oplosmiddel 1 maand-20°Cin oplosmiddel
Synoniemen	N/A			Smiles	CCCS(=O)(=O)NC1=C(C(=C(C=C1)F)C(=O)C2=CNC3=C2C=C(C=N3)Cl)F

Oplosbaarheid

In vitro
Batch:

DMSO : 83 mg/mL (200.56 mM)
(Met vocht verontreinigde DMSO kan de oplosbaarheid verminderen. Gebruik verse, watervrije DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molariteitscalculator

Massa	Concentratie	Volume	Moleculair gewicht
=	×	×

Verdunningscalculator Moleculair gewicht calculator

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.

In vivo Formuleringscalculator (Heldere oplossing)

Stap 1: Voer de onderstaande informatie in (Aanbevolen: Een extra dier voor het geval van verlies tijdens het experiment)

Dosering: mg/kg Gemiddeld gewicht van dieren: g Doseervolume per dier:μL Aantal dieren:

Stap 2: Voer de in vivo formulering in (Dit is alleen de calculator, geen formulering. Neem eerst contact met ons op als er geen in vivo formulering is in het gedeelte Oplosbaarheid.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Berekeningsresultaten:

Werkconcentratie: mg/ml;

Methode voor het bereiden van DMSO-mastervloeistof: mg geneesmiddel vooraf opgelost in μL DMSO ( Concentratie mastervloeistof mg/mL, Neem eerst contact met ons op als de concentratie de DMSO-oplosbaarheid van de partij geneesmiddel overschrijdt. )

Methode voor het bereiden van in vivo formulering: Neem μL DMSO mastervloeistof, voeg vervolgens toeμL PEG300, mengen en helder maken, voeg vervolgens toeμL Tween 80, mengen en helder maken, voeg vervolgens toe μL ddH₂O, mengen en helder maken.

Methode voor het bereiden van in vivo formulering: Neem μL DMSO mastervloeistof, voeg vervolgens toe μL Maïsolie, mengen en helder maken.

Opmerking: 1. Zorg ervoor dat de vloeistof helder is voordat u het volgende oplosmiddel toevoegt.
2. Zorg ervoor dat u het/de oplosmiddel(en) in de juiste volgorde toevoegt. U moet ervoor zorgen dat de verkregen oplossing, bij de vorige toevoeging, een heldere oplossing is voordat u verdergaat met het toevoegen van het volgende oplosmiddel. Fysische methoden zoals vortexen, echografie of een warmwaterbad kunnen worden gebruikt om het oplossen te bevorderen.

Werkingsmechanisme

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 vertoont >10 keer selectiviteit tegen wild-type B-Raf en >100 keer selectiviteit ten opzichte van andere kinasen zoals Frk, Src, Fak, FGFR en Aurora A met IC50 van 1,3-3,4 μM. Deze verbinding remt de ERK-fosforylering significant in cellijnen die B-Raf^V600E bevatten met IC50 van 14-46 nM, maar niet in cellen met wild-type B-Raf. Het remt significant de groei van tumorcellijnen die het B-Raf^V600E-oncogen bevatten, zoals COLO205, A375, WM2664 en COLO829 met GI50 van respectievelijk 0,31 μM, 0,50 μM, 1,5 μM en 1,7 μM. Bovendien induceert deze chemische behandeling bij 1 μM celcyclusarrest en apoptose uitsluitend in de B-Raf^V600E-positieve 1205Lu-cellen, maar niet in de B-Raf wild-type C8161-cellen. Deze verbindingbehandeling (10 μM) induceert significant >14-voudige expressie van BIM in de PTEN⁺-cellen, vergeleken met de PTEN- cellijnen (4-voudig), wat een verklaring geeft voor de resistentie van PTEN^--cellen tegen deze chemisch geïnduceerde apoptose.
Kinase Assay	In vitro Raf kinase-activiteiten
Kinase Assay	De in vitro kinase-activiteiten van wild-type Raf en mutanten worden bepaald door de fosforylering van gebiotinyleerd-MEK-eiwit te meten met behulp van Perkin-Elmer's AlphaScreen-technologie. Voor elk enzym (0,1 ng) worden reacties van 20 μL uitgevoerd in 20 mM Hepes (pH 7,0), 10 mM MgCl₂, 1 mM DTT, 0,01% Tween-20, 100 nM biotine-MEK-eiwit, verschillende ATP-concentraties en toenemende concentraties PLX-4720 bij kamertemperatuur. Reacties worden gestopt na 2, 5, 8, 10, 20 en 30 minuten met 5 μL van een oplossing die 20 mM Hepes (pH 7,0), 200 mM NaCl, 80 mM EDTA en 0,3% BSA bevat. De stopoplossing bevat ook phospho-MEK-antilichaam, met Streptavidin gecoate Donorkralen en Proteïne A Acceptorkralen uit de AlphaScreen Proteïne A Detectiekit. Het antilichaam en de kralen worden 30 minuten vooraf geïncubeerd in de stopoplossing in het donker bij kamertemperatuur. De uiteindelijke verdunning van het antilichaam is 1/2.000, en de uiteindelijke concentratie van elke kraal is 10 μg/mL. De assayplaten worden een uur bij kamertemperatuur geïncubeerd en vervolgens afgelezen op een PerkinElmer AlphaQuest-lezer.
In vivo	Orale toediening van PLX-4720 met 20 mg/kg/dag induceert significante vertragingen in tumorgroei en regressies in B-Raf^V600E-afhankelijke COLO205-tumorxenografts, zonder duidelijke bijwerkingen bij muizen, zelfs bij een dosis van 1 g/kg. Deze verbinding bij 100 mg/kg tweemaal daags elimineert bijna volledig de 1205Lu-xenografts die B-Raf^V600E bevatten, terwijl het geen activiteit heeft tegen C8161-xenografts die wild-type B-Raf bevatten. De anti-tumoreffecten van deze verbinding correleren met de blokkade van de MAPK-route in die cellen die de V600E-mutatie bevatten. Deze chemische behandeling bij 30 mg/kg/dag remt de tumorgroei van 8505c-xenografts significant met >90%, en vermindert dramatisch verre longmetastasen.
Referenties	[1] https://pubmed.ncbi.nlm.nih.gov/18287029/ [2] https://pubmed.ncbi.nlm.nih.gov/21317224/ [3] https://pubmed.ncbi.nlm.nih.gov/20498063/

Toepassingen

Methoden	Biomarkers	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

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

Antwoord:
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.

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