Home Metabolism Lipase inhibitor Orlistat

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Orlistat Lipase inhibitor

Cat.nr.: S1629

Orlistat is a general lipase inhibitor with IC50 of 122 ng/ml for PL from human duodenal juice. This compound treatment reduces proliferation, induces apoptosis and arrests cell cycle.
Orlistat Lipase inhibitor Chemical Structure

Chemische structuur

Molecuulgewicht: 495.73

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Kwaliteitscontrole (Quality Control)

Batch: Zuiverheid: 99.98%
99.98

Celkweek, behandeling & werkzame concentratie
(Cell Culture, Treatment & Working Concentration)

Cellijnen Assaytype Concentratie Incubatietijd Formulering Activiteitsbeschrijving PMID
HEK293T Function assay Inhibition of human DAGLalpha expressed in HEK293T cell membrane using [14C]SAG substrate in detergent free solution by FRET assay, IC50=0.01 μM 22738638
COS Function assay 15 mins Inhibition of human recombinant DAGLalpha expressed in African green monkey COS cells using sn-1-stearoyl-2-[14C]-arachidonoyl-glycerol as substrate incubated for 15 mins by beta counting analysis, IC50 = 0.001 μM. 26917221
COS7 Function assay 20 mins Inhibition of recombinant human HL expressed in COS7 cells using PED-A1 containing DMPG vesicles as substrate pretreated for 20 mins followed by substrate addition and measured every 20 secs for 10 mins by fluorescence assay, IC50 = 0.003 μM. 30613337
HT1080 Function assay 20 mins Inhibition of endothelial lipase in human HT1080 cells using PED-A1 containing DMPG vesicles as substrate pretreated for 20 mins followed by substrate addition and measured every 20 secs for 10 mins by fluorescence assay, IC50 = 0.006 μM. 30613337
HEK293F Function assay 20 mins Inhibition of recombinant human PL expressed in HEK293F cells using PED-A1 containing DMPG vesicles as substrate pretreated for 20 mins followed by substrate addition and measured every 20 secs for 10 mins by fluorescence assay, IC50 = 0.006 μM. 30613337
HEK293 Function assay 1 uM 10 mins Inhibition of human ABHD6 expressed in HEK293 cells at 1 uM pre-incubated for 10 mins before 2-AG substrate addition followed by rapid 40 fold compound dilution measured after 10 mins by HPLC method, IC50 = 0.01318 μM. 26344596
HEK293 Function assay 1 uM 10 mins Inhibition of human ABHD6 expressed in HEK293 cells at 1 uM pre-incubated for 10 mins before 2-AG substrate addition followed by rapid 40 fold compound dilution measured after 30 mins by HPLC method, IC50 = 0.01413 μM. 26344596
HEK293 Function assay 1 uM 10 mins Inhibition of human ABHD6 expressed in HEK293 cells at 1 uM pre-incubated for 10 mins before 2-AG substrate addition followed by rapid 40 fold compound dilution measured after 60 mins by HPLC method, IC50 = 0.01995 μM. 26344596
N18TG2 Function assay 20 mins Inhibition of DAGLalpha in mouse N18TG2 cells assessed as inhibition of ionomycin-induced formation of 2-AG incubated for 20 mins by LC-MS analysis, IC50 = 0.02 μM. 26917221
HEK293T Function assay Inhibition of recombinant BAT5 transfected in HEK293T cells by SDS-PAGE using rhodamine-tagged FP probe, IC50 = 0.03 μM. 18657971
HEK293 Function assay 1 uM 10 mins Inhibition of human ABHD6 expressed in HEK293 cells at 1 uM pre-incubated for 10 mins before 2-AG substrate addition followed by rapid 40 fold compound dilution measured after 90 mins by HPLC method, IC50 = 0.03715 μM. 26344596
HEK293 Function assay Inhibition of human ABHD6 containing pCMV6-AC-hABHD6 transfected into HEK293 cells, IC50 = 0.04786 μM. 25752982
HEK293 Function assay Inhibition of human ABHD6 containing pCMV6-AC-hABHD6 transfected into HEK293 cells, IC50 = 0.048 μM. 25752982
HEK293T Function assay Inhibition of recombinant PLA2g7 transfected in HEK293T cells by SDS-PAGE using rhodamine-tagged FP probe, IC50 = 0.05 μM. 18657971
COS7 Function assay Inhibition of human recombinant DAGLalpha overexpressed in african green monkey COS7 cells, IC50 = 0.06 μM. 18657971
COS7 Function assay Inhibition of human recombinant DAGLbeta overexpressed in african green monkey COS7 cells, IC50 = 0.06 μM. 18657971
COS7 Function assay 20 mins Inhibition of recombinant human LPL expressed in COS7 cells using PED-A1 containing DMPG vesicles as substrate pretreated for 20 mins followed by substrate addition and measured every 20 secs for 10 mins by fluorescence assay, IC50 = 0.066 μM. 30613337
HEK293T Function assay Inhibition of recombinant ABHD12 transfected in HEK293T cells by SDS-PAGE using rhodamine-tagged FP probe, IC50 = 0.08 μM. 18657971
HEK293 Function assay Inhibition of human ABHD12 containing pCMV6-XL4-hABHD12 transfected into HEK293 cells, IC50 = 0.19 μM. 25752982
HEK293 Function assay 10 mins Inhibition of human ABHD12 expressed in HEK293 cells pre-incubated for 10 mins before 2-AG substrate addition by HPLC method, IC50 = 0.19 μM. 26344596
HEK293 Function assay Inhibition of human ABHD12 containing pCMV6-XL4-hABHD12 transfected into HEK293 cells, IC50 = 0.19055 μM. 25752982
HEK293 Function assay 10 mins Inhibition of human ABHD12 expressed in HEK293 cells pre-incubated for 10 mins before 2-AG substrate addition by HPLC method, IC50 = 0.19055 μM. 26344596
HEK293 Function assay Displacement of [3H]CP-55940 from human recombinant CB1 receptor expressed in HEK293 cells by scintillation counting, Ki = 2.5 μM. 18831576
BxPC3 Function assay 10 to 14 days Inhibition of survival of human BxPC3 cells after 10 to 14 days by crystal violet staining-based colony formation assay, IC50 = 8.45 μM. 25513712
MDA-MB-231 Cytotoxicity assay Cytotoxicity against human MDA-MB-231 cells assessed as reduction in cell viability, IC50 = 13 μM. 29541373
MDA-MB-435 Cytotoxicity assay 48 hrs Cytotoxicity against human MDA-MB-435 cells after 48 hrs by Cell titer assay, IC50 = 16.8 μM. 18710210
HepG2 (DPX-2) Function assay 24 hrs Activation of human PXR expressed in human HepG2 (DPX-2) cells assessed as induction of CYP3A4 after 24 hrs by luminescent analysis, EC50 = 28.2 μM. 20966043
COS7 Function assay Inhibition of recombinant DAGLbeta overexpressed in african green monkey COS7 cells assessed as accumulation of 2-arachidonoylglycerol by Western blotting 18657971
COS7 Function assay Inhibition of recombinant DAGLalpha overexpressed in african green monkey COS7 cells assessed as accumulation of 2-arachidonoylglycerol by Western blotting 18657971
MDA-MB-435 Apoptosis assay 25 uM 72 hrs Induction of apoptosis in human MDA-MB-435 cells assessed as DNA fragmentation at 25 uM after 72 hrs 18710210
HEK293 Function assay 1 uM 10 mins Reversible inhibition of human ABHD6 expressed in HEK293 cells at 1 uM pre-incubated for 10 mins before 2-AG substrate addition followed by rapid 40 fold compound dilution measured after 10 to 90 mins by HPLC method 26344596
LNCAP Function assay 20 uM 48 hrs Induction of cholesterol metabolism deregulation in human LNCAP cells assessed as reduction in NBD-cholesterol uptake at 20 uM after 48 hrs by DAPI/Alexa fluor 633 phalloidin staining based high-content imaging analysis 29474071
BV2 Function assay 30 mins Inhibition of ABHD6 in mouse BV2 cells preincubated for 30 mins and subsequent addition of [3H]-2-OG substrate measured after 15 mins by liquid scintillation counting method 28284861
BV2 Function assay 30 mins Inhibition of ABHD12 in mouse BV2 cells preincubated for 30 mins and subsequent addition of [3H]-2-OG substrate measured after 15 mins by liquid scintillation counting method 28284861
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Chemische informatie, opslag en stabiliteit (Chemical Information, Storage & Stability)

Molecuulgewicht 495.73 Formule

C29H53NO5

 Opslag (vanaf de datum van ontvangst)
CAS-nr. 96829-58-2 SDF downloaden Opslag van stamoplossingen

Synoniemen Tetrahydrolipstatin,Ro 18-0647 Smiles CCCCCCCCCCCC(CC1C(C(=O)O1)CCCCCC)OC(=O)C(CC(C)C)NC=O

Oplosbaarheid (Solubility)

In vitro
Batch:

DMSO : 99 mg/mL (199.7 mM)
(Met vocht verontreinigd DMSO kan de oplosbaarheid verminderen. Gebruik verse, watervrije DMSO.)

Ethanol : 99 mg/mL

Water : Insoluble

Molariteitscalculator

Massa Concentratie Volume Molecuulgewicht
Verdunningscalculator Molecuulgewichtcalculator

In vivo
Batch:

In vivo formulatiecalculator (heldere oplossing)

Stap 1: Voer onderstaande informatie in (Aanbevolen: een extra dier om rekening te houden met verlies tijdens het experiment)

mg/kg g μL

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 de sectie oplosbaarheid.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Berekeningsresultaten:

Werkconcentratie: mg/ml;

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

Methode voor het bereiden van in vivo formulering: Neem μL DMSO moedervloeistof, voeg daarna toeμL PEG300, mengen en verhelderen, daarna toevoegenμL Tween 80, mengen en verhelderen, daarna toevoegen μL ddH2O, mengen en verhelderen.

Methode voor het bereiden van in vivo formulering: Neem μL DMSO moedervloeistof, voeg daarna toe μL Maïsolie, mengen en verhelderen.

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. Fysieke methoden zoals vortexen, ultrasoon of een warmwaterbad kunnen worden gebruikt om het oplossen te bevorderen.

Werkingsmechanisme (Mechanism of Action)

Targets/IC50/Ki
lipase
(Cell-free assay)
Fatty acid synthesis
(Cell-free assay)
In vitro
Orlistat, een remmer van lipasen en Fatty Acid Synthase, wordt oraal gebruikt voor de langetermijnbehandeling van obesitas. Deze verbinding vertoont antiproliferatieve activiteit tegen kankercellen in vitro. Er is vastgesteld dat het pro-apoptosis NOXA-eiwit verhoogt.
In vivo
Orlistat, oraal toegediend, wordt minimaal geabsorbeerd door het maagdarmkanaal en is in staat de absorptie van een groot percentage lipiden te voorkomen, waardoor de lipidetoevoer van externe bronnen wordt verminderd. Vanwege de extreem lage orale biologische beschikbaarheid zijn de effecten van deze verbinding grotendeels beperkt tot het maagdarmkanaal, waar het pancreaslipase inactiveert. Daarom zouden de formulering en toedieningsroute moeten worden gewijzigd om tumoren van de borst, prostaat, enzovoort te behandelen. Deze chemische stof stopt de proliferatie van tumorcellen, induceert tumorcel apoptosis en remt de groei van PC-3-tumoren bij naakte muizen. Een farmacokinetische analyse van deze verbinding (155 mg/kg) toegediend via i.p. injectie toonde piekniveaus in het bloed van ∼10 μM 2 uur na dosering (gegevens niet getoond). Na deze tijd namen de bloedspiegels van het medicijn snel af.
Referenties

Toepassingen (Applications)

Methoden Biomarkers Afbeeldingen PMID
Western blot FASN / AR / p-AKT / p-p53 / p53 / VEGF / Cyclin D1 / Bcl-2 / Cleaved caspase-3
S1629-WB1
31527721
Growth inhibition assay Cell viability
S1629-viability1
28387458

Informatie over klinische proeven (Clinical Trial Information)

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

NCT-nummer Werving Aandoeningen Sponsor/medewerkers Startdatum Fasen
NCT01755676 Completed
Obesity
EMS
 September 2016 Phase 3
NCT02141230 Withdrawn
Weight Loss
GlaxoSmithKline|Hamell
 December 2015 Not Applicable
NCT01719419 Withdrawn
Overweight
Pennington Biomedical Research Center
 March 2012 Not Applicable
NCT01332448 Completed
Obesity
GlaxoSmithKline
 February 2010 --
NCT01414465 Completed
Overweight
University of Campinas Brazil|Germed Pharma
 October 2009 Not Applicable