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PR-171 (Carfilzomib) Proteasome remmer

Name: PR-171 (Carfilzomib)
Brand: Selleck Chemicals
SKU: S2853
Rating: 5 (249 reviews)

Cat.Nr.S2853

Carfilzomib (PR-171) is een irreversibele proteasome remmer met een IC50 van <5 nM in ANBL-6 cellen, toonde een preferentiële in vitro remmende potentie tegen de ChT-L activiteit in de β5-subeenheid, maar weinig of geen effect op de PGPH en T-L activiteiten. Carfilzomib activeert pro-overleving autophagy en induceert cel apoptosis.

PR-171 (Carfilzomib) Proteasome remmer Chemical Structure

Chemische structuur

Moleculair gewicht: 719.91

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Kwaliteitscontrole

Batch: Zuiverheid: 99.75%

99.75

Gerelateerde doelwitten	HDAC Caspase Secretase MMP HCV Protease Cysteine Protease DPP Tyrosinase HIV Protease Serine Protease
Overige Proteasome Inhibitoren	MG132 Celastrol Epoxomicin (BU-4061T) ONX-0914 (PR-957) Oprozomib Delanzomib VR23 Marizomib (Salinosporamide A) PI-1840 KSQ-4279 (USP1-IN-1)

Celkweek, behandeling & werkconcentratie

Cellijnen	Assaytype	Concentratie	Incubatietijd	Activiteitsbeschrijving	PMID
MM.1S	Growth Inhibition Assay	0-100 nM	48 h	IC50 = 10 nM	25312543
NCI-H929	Growth Inhibition Assay	0-100 nM	48 h	IC50 = 14 nM	25312543
SUDHL16	Apoptosis Asssay	2.5–3.5 nM	48 h	enhances the cell death co-treatment with ACY1215	25239935
SUDHL14	Apoptosis Asssay	2.5–3.5 nM	48 h	enhances the cell death co-treatment with ACY1215	25239935
U2932	Apoptosis Asssay	2.5–3.5 nM	48 h	enhances the cell death co-treatment with ACY1215	25239935
P-UMSCC-1	Growth Inhibition Assay			IC50=11.2 nM	24915039
R-UMSCC-1	Growth Inhibition Assay			IC50=2294 nM	24915039
P-Cal33	Growth Inhibition Assay			IC50=17.3 nM	24915039
R-Cal33	Growth Inhibition Assay			IC50=1112 nM	24915039
Jurkat	Growth Inhibition Assay	1-11nM	48 h	inhibits the cell proliferation co-treatment with vorinostat	24801128
Jurkat	Apoptosis Asssay	8 nM	24/48 h	induces apoptosis, caspase activation, and PARP cleavage co-treatment with vorinostat	24801128
UMSCC-22A	Apoptosis Asssay	200 nM	24 h	induce the cell apoptosis co-treatment with ONX 0912	22929803
UMSCC-22B	Apoptosis Asssay	200 nM	24 h	induce the cell apoptosis co-treatment with ONX 0912	22929803
1483	Apoptosis Asssay	200 nM	24 h	induce the cell apoptosis co-treatment with ONX 0912	22929803
UMSCC-1	Apoptosis Asssay	200 nM	24 h	induce the cell apoptosis co-treatment with ONX 0912	22929803
UMSCC-22A	Growth Inhibition Assay			IC50=38.7 ± 1.0 nM	22929803
UMSCC-22B	Growth Inhibition Assay			IC50=30.7 ± 9.3 nM	22929803
1483	Growth Inhibition Assay			IC50=50.5 ± 11.9 nM	22929803
UMSCC-1	Growth Inhibition Assay			IC50=34.6 ± 2.6 nM	22929803
Cal33	Growth Inhibition Assay			IC50=49.3 ± 8.9 nM	22929803
PCI-15A	Growth Inhibition Assay			IC50=70.4 ± 22.6 nM	22929803
PCI-15B	Growth Inhibition Assay			IC50=39.5 ± 11.0 nM	22929803
OSC-19	Growth Inhibition Assay			IC50=18.3 ± 4.2 nM	22929803
SUDHL16	Apoptosis Asssay	2.0-4.0 nM	48 h	induces cell death co-treatment with obatoclax	22411899
SUDHL16	Function Assay	2.5 nM	24 h	activates JNK, inactivates AKT, up-regulates Noxa, and induces γH2A.X co-treatment with obatoclax	22411899
Granta	Growth Inhibition Assay	0-4 nM	48 h	induce cell death co-treatment with HADCIs	21750224
SUDHL16	Growth Inhibition Assay	1-4 nM	36 h	induce cell death co-treatment with HADCIs	20233973
MOLT4	Function assay		1 hr	Inhibition of chymotrypsin-like activity of 20S proteasome in human MOLT4 cells after 1 hr by CellTiter-Glo luminescent assay, IC50 = 0.0051 μM.	19348473
MESSA	Cytotoxicity assay		72 hrs	Cytotoxicity against human MESSA cells assessed as cell viability after 72 hrs by CellTiter-Glo luminescent assay, IC50 = 0.018 μM.	19348473
MESSA	Cytotoxicity assay		72 hrs	Cytotoxicity against multidrug resistance transporter expressing doxorubicin resistant human MESSA cells assessed as cell viability after 72 hrs by CellTiter-Glo luminescent assay, IC50 = 0.413 μM.	19348473
RPMI8226	Cytotoxicity assay		72 hrs	Cytotoxic activity against human RPMI8226 cells after 72 hrs by MTS assay, IC50 = 0.01319 μM.	24767818
NCI-H929	Cytotoxicity assay		72 hrs	Cytotoxic activity against human NCI-H929 cells after 72 hrs by MTS assay, IC50 = 0.02132 μM.	24767818
CCRF-CEM	Antiproliferative assay		72 hrs	Antiproliferative activity against human CCRF-CEM cells after 72 hrs by oxyluciferin luminescence assay, IC50 = 0.0061 μM.	26231162
RPMI8266	Antiproliferative assay		72 hrs	Antiproliferative activity against human RPMI8266 cells after 72 hrs by oxyluciferin luminescence assay, IC50 = 0.0139 μM.	26231162
HCT116	Antiproliferative assay		72 hrs	Antiproliferative activity against human HCT116 cells after 72 hrs by oxyluciferin luminescence assay, IC50 = 0.0193 μM.	26231162
A431	Antiproliferative assay		72 hrs	Antiproliferative activity against human A431 cells after 72 hrs by oxyluciferin luminescence assay, IC50 = 0.0238 μM.	26231162
TOV21G	Antiproliferative assay		72 hrs	Antiproliferative activity against human TOV21G cells after 72 hrs by oxyluciferin luminescence assay, IC50 = 0.0238 μM.	26231162
RKO	Antiproliferative assay		72 hrs	Antiproliferative activity against human RKO cells after 72 hrs by oxyluciferin luminescence assay, IC50 = 0.0271 μM.	26231162
MM1S	Antiproliferative assay		72 hrs	Antiproliferative activity against human MM1S cells measured after 72 hrs by MTS assay, IC50 = 0.0015 μM.	27765408
RPMI8226	Antiproliferative assay		72 hrs	Antiproliferative activity against human RPMI8226 cells measured after 72 hrs by MTS assay, IC50 = 0.0132 μM.	27765408
LCL	Cytotoxicity assay		48 hrs	Cytotoxicity against human LCL cells harboring wild type p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.03 μM.	27994734
RD-ES	Cytotoxicity assay		48 hrs	Cytotoxicity against human RD-ES cells harboring p53 mutant assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.043 μM.	27994734
U266	Cytotoxicity assay		48 hrs	Cytotoxicity against human U266 cells harboring mutant p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.06 μM.	27994734
WE68	Cytotoxicity assay		48 hrs	Cytotoxicity against human WE68 cells harboring wild type p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.08 μM.	27994734
IMR90	Cytotoxicity assay		48 hrs	Cytotoxicity against human IMR90 cells harboring wild type p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.13 μM.	27994734
MCF10A	Cytotoxicity assay		48 hrs	Cytotoxicity against human MCF10A cells harboring wild type p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.32 μM.	27994734
SKOV3	Cytotoxicity assay		48 hrs	Cytotoxicity against p53 deficient human SKOV3 cells assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.32 μM.	27994734
MDA-MB-468	Cytotoxicity assay		48 hrs	Cytotoxicity against human MDA-MB-468 cells harboring mutant p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.33 μM.	27994734
HNDF	Cytotoxicity assay		48 hrs	Cytotoxicity against HNDF cells harboring wild type p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.35 μM.	27994734
KGN	Cytotoxicity assay		48 hrs	Cytotoxicity against human KGN cells harboring wild type p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 0.45 μM.	27994734
MCF7	Cytotoxicity assay		48 hrs	Cytotoxicity against human MCF7 cells harboring wild type p53 assessed as reduction in cell viability after 48 hrs by 7AAD-staining based FACS analysis, LD50 = 4.5 μM.	27994734
MCF7	Function assay	35 nM	4 hrs	Inhibition of 26S proteasome in human MCF7 cells assessed as accumulation of high molecular weight polyubiquitin-conjugated proteins at 35 nM after 4 hrs by Western blot analysis	27994734
MDA-MB-468	Function assay	35 nM	4 hrs	Inhibition of 26S proteasome in human MDA-MB-468 cells assessed as accumulation of high molecular weight polyubiquitin-conjugated proteins at 35 nM after 4 hrs by Western blot analysis	27994734
MM1S	Cytotoxicity assay		72 hrs	Cytotoxicity against human MM1S cells measured after 72 hrs by MTS assay, IC50 = 0.0015 μM.	28027531
RPMI8226	Cytotoxicity assay		72 hrs	Cytotoxicity against human RPMI8226 cells measured after 72 hrs by MTS assay, IC50 = 0.0132 μM.	28027531
TC32	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for TC32 cells	29435139
U-2 OS	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for U-2 OS cells	29435139
A673	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
DAOY	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY 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
BT-37	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 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
BT-12	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells	29435139
MG 63 (6-TG R)	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for MG 63 (6-TG R) 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
BT-12	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-12 cells	29435139
LAN-5	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for LAN-5 cells	29435139
fibroblast cells	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for control Hh wild type fibroblast cells	29435139
NB-EBc1	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for NB-EBc1 cells	29435139
SK-N-SH	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-SH cells	29435139
Rh41	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells	29435139
A673	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for A673 cells)	29435139
Rh30	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh30 cells	29435139
BT-37	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-37 cells	29435139
MG 63 (6-TG R)	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for MG 63 (6-TG R) cells	29435139
Rh30	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh30 cells	29435139
fibroblast cells	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for control Hh wild type fibroblast cells	29435139
OHS-50	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for OHS-50 cells	29435139
SK-N-SH	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for SK-N-SH cells	29435139
Daoy	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for Daoy cells	29435139
TC32	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D caspase screen for TC32 cells	29435139
TC32	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for TC32 cells	29435139
MG 63 (6-TG R)	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for MG 63 (6-TG R) 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
NB-EBc1	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells	29435139
LAN-5	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for LAN-5 cells	29435139
Rh18	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh18 cells	29435139
SK-N-MC	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-MC cells	29435139
SJ-GBM2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SJ-GBM2 cells	29435139
TC32	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for TC32 cells	29435139
Rh18	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh18 cells	29435139
Saos-2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Saos-2 cells	29435139
SJ-GBM2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for SJ-GBM2 cells	29435139
RD	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for RD cells	29435139
ANBL-6	Function assay			Inhibition of 20S proteasome activity in human ANBL-6 cells, IC50 = 0.01 μM.	29652143
MCF7	Cytotoxicity assay		48 hrs	Cytotoxicity against human MCF7 cells after 48 hrs by MTT assay, IC50 = 0.0041 μM.	30165344
MDA-MB-231	Cytotoxicity assay		48 hrs	Cytotoxicity against human MDA-MB-231 cells after 48 hrs by MTT assay, IC50 = 0.0044 μM.	30165344
RPMI8226	Cytotoxicity assay		48 hrs	Cytotoxicity against human RPMI8226 cells after 48 hrs by MTT assay, IC50 = 0.0067 μM.	30165344
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Chemische informatie, Opslag en Stabiliteit

Moleculair gewicht	719.91	Formule	C₄₀H₅₇N₅O₇	Opslag (Vanaf de ontvangstdatum)	3 jaar-20°Cpoeder
CAS-nr.	868540-17-4	SDF downloaden		Opslag van stamoplossingen	1 jaar-80°Cin oplosmiddel 1 maand-20°Cin oplosmiddel
Synoniemen	N/A			Smiles	CC(C)CC(C(=O)C1(CO1)C)NC(=O)C(CC2=CC=CC=C2)NC(=O)C(CC(C)C)NC(=O)C(CCC3=CC=CC=C3)NC(=O)CN4CCOCC4

Oplosbaarheid

In vitro
Batch:

DMSO : 100 mg/mL (138.9 mM)
(Met vocht verontreinigde DMSO kan de oplosbaarheid verminderen. Gebruik verse, watervrije DMSO.)

Ethanol : 50 mg/mL

Water : 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.
Clear solution	2%DMSO 1 30%PEG300 2 2%TWEEN80 3 66%ddH2O Gevalideerd door Selleck labs. Mocht u aanpassingen aan deze formulering nodig hebben, neem dan contact op met ons verkoopteam voor aangepaste tests.	1.000mg/ml (1.39mM)	Taking the 1 mL working solution as an example, add 20 μL of 50 mg/ml clarified DMSO stock solution to 300 μL of PEG300, mix evenly to clarify it; add 20 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 660 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
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	2%DMSO 1 30%PEG300 2 2%Tween80 3 66%ddH2O Gevalideerd door Selleck labs. Mocht u aanpassingen aan deze formulering nodig hebben, neem dan contact op met ons verkoopteam voor aangepaste tests.	1.000mg/ml (1.39mM)	Taking the 1 mL working solution as an example, add 20 μL of 50 mg/ml clarified DMSO stock solution to 300 μL of PEG300, mix evenly to clarify it; add 20 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 660 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
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	Proteasome (ANBL-6 cells) 5 nM
In vitro	Carfilzomib (PR-171) remt de proliferatie in een verscheidenheid aan cellijnen en van patiënten afgeleide neoplastische cellen, waaronder multipel myeloom, en induceerde intrinsieke en extrinsieke apoptotische signaalroutes en activering van c-Jun-N-terminale kinase (JNK). Het onthult een verbeterde anti-MM-activiteit, overwint resistentie tegen andere middelen en werkt synergetisch met (Dex). Deze verbinding vertoont een preferentiële in vitro remmende potentie tegen de ChT-L-activiteit in de β5-subeenheid, met meer dan 80% remming bij doses van 10 nM. Korte blootstelling aan een lage dosis Carfilzomib leidt tot preferentiële bindingsspecificiteit voor het β5 constitutieve 20S proteasome en de β5i immunoproteasome-subeenheden. Meting van caspase-activiteit in ANBL-6-cellen gepulst ermee toont aanzienlijke stijgingen van caspase-8-, caspase-9- en caspase-3-activiteit na 8 uur, wat respectievelijk een 3,2-, 3,9- en 6,9-voudige toename oplevert ten opzichte van controlecellen na 8 uur. In met carfilzomib gepulseerde cellen is de mitochondriale membraanintegriteit gedaald tot 41% (Q1 + Q2), vergeleken met 75% in met vehikel behandelde controlecellen. In een andere studie heeft het ook preklinische effectiviteit getoond tegen hematologische en solide maligniteiten. Het remt direct de osteoclastenformatie en botresorptie.
Kinase Assay	Enzyme-linked immunosorbent assay voor subeenheidsprofilering van carfilzomib
Kinase Assay	ANBL-6-cellen (2 × 10⁶/well) worden geplateerd in 96-wells platen en gedurende 1 uur behandeld met Carfilzomib (PR-171) doses van 0,001 tot 10 μM. Cellen worden vervolgens gelyseerd (20 mM Tris-HCl, 0,5 mM EDTA) en gezuiverde lysaten worden overgebracht naar polymeraseketenreactie (PCR) platen. Een standaardcurve wordt gegenereerd met onbehandelde ANBL-6-cellysaten, beginnend bij een concentratie van 6 μg eiwit/μL. De actieve-plaats-probe [biotin-(CH2)4-Leu-Leu-Leu-epoxyketon; 20 μM] wordt toegevoegd en gedurende 1 uur geïncubeerd bij kamertemperatuur. Cellysaten worden vervolgens gedenatureerd door 1% natriumdodecylsulfaat (SDS) toe te voegen en te verwarmen tot 100°C, gevolgd door menging met 20 μL per well streptavidine-sepharose high-performance parels in een 96-wells multiscreen DV-plaat en gedurende 1 uur geïncubeerd. Deze parels worden vervolgens gewassen met enzyme-linked immunosorbent assay (ELISA) buffer (PBS, 1% runderserumalbumine en 0,1% Tween-20) en 's nachts bij 4°C geïncubeerd op een plaatschudder met antilichamen tegen proteasome-subeenheden. Gebruikte antilichamen omvatten muismonoklonaal anti-β1, anti-β2, anti-β1i en anti-β5i, geit-polyklonaal anti-β2i en konijn-polyklonaal anti-β5 (affiniteit-gezuiverd antiserum tegen KLH-CWIRVSSDNVADLHDKYS-peptide). De parels worden gewassen en gedurende 2 uur geïncubeerd met mierikswortelperoxidase-geconjugeerde secundaire geit-antikonijn, geit-antimuis of konijn-antigeit antilichamen. Na het wassen worden de parels ontwikkeld met behulp van het supersignal ELISA picochemiluminescentie substraat. Luminescente detectie wordt uitgevoerd. Ruwe luminescentie wordt omgezet naar μg/mL door vergelijking met de standaardcurve en uitgedrukt als de % remming ten opzichte van de voertuigcontrole. Curvefits worden gegenereerd met behulp van de volgende niet-sigmoïdale dosis-responsvergelijking: Y = Onderkant + (Bovenkant-Onderkant)/(1 + 10̂((LogEC50 − X) × HillSlope)), waarbij X het logaritme van de concentratie is, Y de % remming, en EC50 de dosis van deze verbinding die 50% effect vertoont.
In vivo	Carfilzomib (PR-171) vermindert de tumorgroei matig in een in vivo xenograftmodel. Het vermindert effectief de levensvatbaarheid van multipel myeloomcellen na continue of transiënte nabootsende behandeling. Deze verbinding verhoogt ook het trabeculair botvolume, vermindert botresorptie en verbetert de botvorming bij niet-tumor dragende muizen.
Referenties	[1] https://pubmed.ncbi.nlm.nih.gov/17591945/ [2] https://pubmed.ncbi.nlm.nih.gov/21247387/ [3] https://pubmed.ncbi.nlm.nih.gov/22763387/ [4] https://pubmed.ncbi.nlm.nih.gov/21750224/

Toepassingen

Methoden	Biomarkers	Afbeeldingen	PMID
Western blot	pERK / ERK / pSTAT5 / STAT5 / pPI3K / PI3K caspase-9 / caspase-8 c-PARP / PARP / caspase-3 Bcl-2 / Bcl-Xl / Mcl-1 / Bik / Bim / Bax / Bak Atg5 / Atg12 / Beclin-1 / LC3-II Noxa / Bik / Puma / Mcl-1 EGFR / HER2 / ER alpha / p-Akt(Ser473) / Akt / p-ERK / ERK / p53 BDP1 / HER2(Tyr1248) / HER2(Tyr1221/Tyr1222) / PARP1 / caspase-7 / p53 Mut HLA class I		24590311
Growth inhibition assay	Cell viability		27655642

Informatie klinische proef

(gegevens van https://clinicaltrials.gov, bijgewerkt op 2024-05-22)

NCT-nummer	Rekrutering	Aandoeningen	Sponsor/Medewerkers	Startdatum	Fasen
NCT05552976	Recruiting	Relapsed or Refractory Multiple Myeloma	Bristol-Myers Squibb	January 10 2023	Phase 3
NCT05675449	Recruiting	Multiple Myeloma	Pfizer	December 14 2022	Phase 1
NCT05041933	Unknown status	Hematological Diseases	University Hospital Limoges	September 15 2021	--

Technische ondersteuning

Gebruiksaanwijzing

Tel: +1-832-582-8158 Ext:3

Als u nog andere vragen heeft, kunt u een bericht achterlaten.

Veelgestelde vragen

Vraag 1:
How should I prepare a solution of it for an ongoing in vivo study?

Antwoord:
It can be dissolved in 2% DMSO/30% PEG 300/dd H₂O at 10 mg/ml as a suspension, and can be dissolved in 2% DMSO/castor oil at 10 mg/ml as a clear solution.

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):