Osimertinib (AZD9291)

Synonyms: Mereletinib

Osimertinib (AZD9291) is an oral, irreversible, and mutant-selective EGFR inhibitor with IC50 of 12.92, 11.44 and 493.8 nM for Exon 19 deletion EGFR, L858R/T790M EGFR, and WT EGFR in LoVo cells, respectively. Phase 3.

Osimertinib (AZD9291) Chemical Structure

Osimertinib (AZD9291) Chemical Structure

CAS: 1421373-65-0

Selleck's Osimertinib (AZD9291) has been cited by 361 publications

Purity & Quality Control

Batch: Purity: 99.99%
99.99

Products often used together with Osimertinib (AZD9291)

Afatinib


Both are EGFR inhibitors. Their combination therapy induces a more potent inhibitory effect against several EGFR exon 20 insertion mutations than either therapy alone.

Hasegawa H, et al. Lung Cancer. 2019 Jan;127:146-152.

Gefitinib


Both are EGFR inhibitors. Their combination therapy is used in treating of Non-small cell lung cancer in patients harboring the T790M and C797S mutations.

Arulananda S, et al. J Thorac Oncol. 2017 Nov;12(11):1728-1732.

Erlotinib


Both are EGFR inhibitors. Osmiertinib is used in combination with erlotinib in patients with resistance to first and second generation TKIs in treatment of non-small cell lung carcinoma

Trametinib (GSK1120212)


Osimertinib and Trametinib along with dabrafinib can overcome BRAF V600E–induced osimertinib resistance in patients bearing BRAF V600E, EGFR 19del, and T790M mutations

Huang Y, et al. Journal of Thoracic Oncology 14.10 (2019): e236-e237.

Lazertinib


Lazertinib in combination with Amivantamab are used for the treatment of osimertinib-relapsed, chemotherapy-naïve EGFR mutant (EGFRm) non-small cell lung cancer. It is also used as potenial biomarker for response

Bauml J, et al. Journal of Clinical Oncology 39, no. 15_suppl (May 20, 2021) 9006-9006.

Osimertinib (AZD9291) Related Products

Signaling Pathway

Choose Selective EGFR Inhibitors

Cell Data

Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
PC-9/BRc1 Function Assay 50 nM 24 h DMSO induces expression of the proapoptotic BCL-2 family member BIM 25477325
PC-9/ERc1 Function Assay 50 nM 24 h DMSO induces expression of the proapoptotic BCL-2 family member BIM 25477325
VP-2 Function Assay 50 nM 24 h DMSO induces expression of the proapoptotic BCL-2 family member BIM 25477325
PC-9/BRc1 Growth Inhibition Assay 50 nM 10 d DMSO inhibits proliferation in long-term (10-day) growth inhibition assays 25477325
PC-9/ERc1 Growth Inhibition Assay 50 nM 10 d DMSO inhibits proliferation in long-term (10-day) growth inhibition assays 25477325
VP-2 Growth Inhibition Assay 50 nM 10 d DMSO inhibits proliferation in long-term (10-day) growth inhibition assays 25477325
PC9 GR4 Function Assay 0-10 μM 72 h inhibits EGFR phosphorylation and downstream signaling  25948633
PC9 Function Assay 0-10 μM 72 h inhibits WT EGFR at low concentrations 25948633
PC9 GR4 Growth Inhibition Assay 0-10 μM 72 h inhibits cell growth dose dependently 25948633
BAF3 Function assay 72 h GI50 = 0.0003 μM 28282122
BAF3 Function assay 72 h GI50 = 0.0003 μM 28282122
BAF3 Function assay 72 h GI50 = 0.001 μM 28282122
HCC827 Function assay 72 h GI50 = 0.001 μM 28282122
PC9 Function assay 72 h GI50 = 0.002 μM 28282122
BAF3 Function assay 4 h EC50 = 0.002 μM 28282122
HCC827 Function assay 3 h IC50 = 0.0025 μM 27433829
H1975 Function assay 3 h IC50 = 0.0025 μM 27433829
H3255 Function assay 3 h IC50 = 0.0041 μM 27433829
NCI-H1975 Function assay 72 h GI50 = 0.005 μM 28282122
PC9 Antiproliferative activity assay 72 h IC50 = 0.0065 μM 28716641
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.0105 μM 28716641
Sf21 Function assay IC50 = 0.012 μM 27996267
PC9-DRH Function assay 2 h IC50 = 0.013 μM 26756222
BAF3 Function assay 72 h GI50 = 0.013 μM 28282122
HCC827 Function assay 96 h EC50 = 0.014 μM 28225269
HCC827 Antiproliferative activity assay 96 h EC50 = 0.014 μM 28853575
NCI-H1975 Antiproliferative activity assay 96 h EC50 = 0.014 μM 28853575
NCI-H1975 Function assay 2 h IC50 = 0.015 μM 26756222
H1975 Function assay 2 h IC50 = 0.015 μM 26968253
PC9 Function assay 2 h IC50 = 0.017 μM 26968253
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.019 μM 29466773
NCI-H1975 Function assay 96 h EC50 = 0.019 μM 28225269
NCI-H1975 Antiproliferative activity assay 96 h EC50 = 0.019 μM 28603991
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.019 μM 29853340
HCC827 Function assay 2 h IC50 = 0.023 μM 26756222
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.023 μM 29534926
PC9 Cytotoxicity assay 72 h GI50 = 0.023 μM 25271963
NCI-H1975 Cytotoxicity assay 72 h GI50 = 0.024 μM 25271963
HCC827 Antiproliferative activity assay 72 h IC50 = 0.0254 μM 29576272
HCC827 Antiproliferative activity assay 72 h IC50 = 0.027 μM 29466773
HCC827 Antiproliferative activity assay 72 h IC50 = 0.027 μM 29853340
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.03 μM 28033579
H3255 Function assay 72 h GI50 = 0.033 μM 28282122
H3255 Function assay 2 h IC50 = 0.036 μM 26756222
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.041 μM 29730192
NCI-H1975 Function assay 1 h IC50 = 0.041 μM 29534926
BAF3 Function assay 4 h EC50 = 0.043 μM 28282122
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.0472 μM 29576272
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.052 μM 27131639
PC9 Function assay 2 h IC50 = 0.056 μM 26756222
NCI-H1975 Cytotoxicity assay 72 h IC50 = 0.06 μM 29486953
HCC827 Antiproliferative activity assay IC50 = 0.0616 μM 28426996
NCI-H1975 Antiproliferative activity assay IC50 = 0.067 μM 28426996
HaCaT Function assay 3 h IC50 = 0.0737 μM 27433829
NCI-H1975 Antiproliferative activity assay 72 h IC50 = 0.13 μM 30429956
A431 Function assay 1 h IC50 = 0.141 μM 29534926
A549 Function assay IC50 = 0.15 μM 26756222
Calu3 Cytotoxicity assay 72 h GI50 = 0.264 μM 25271963
Sf9 Function assay 20 mins IC50 = 0.278 μM 28482151
BAF3 Function assay 72 h GI50 = 0.3 μM 28282122
BAF3 Function assay 72 h GI50 = 0.31 μM 28282122
NCI-H460 Antiproliferative activity assay 72 h IC50 = 0.4159 μM 28716641
LoVo Function assay 2 h IC50 = 0.48 μM 26968253
LoVo Function assay 2 h IC50 = 0.48 μM 27996267
A549 Antiproliferative activity assay 72 h IC50 = 0.486 μM 29576272
BAF3 Function assay 72 h GI50 = 0.5 μM 28282122
A549 Antiproliferative activity assay 72 h IC50 = 0.53 μM 29466773
A549 Cytotoxicity assay 72 h IC50 = 0.53 μM 29853340
BAF3 Function assay 72 h GI50 = 0.55 μM 28282122
BAF3 Function assay 72 h GI50 = 0.56 μM 28282122
HEK293 Function assay IC50 = 0.57 μM 28426996
BAF3 Function assay 72 h GI50 = 0.59 μM 28282122
A431 Antiproliferative activity assay IC50 = 0.6156 μM 28426996
HT-29 Cytotoxicity assay 72 h IC50 = 0.65 μM 29486953
A431 Function assay 96 h EC50 = 0.667 μM 28225269
A431 Antiproliferative activity assay 96 h EC50 = 0.67 μM 28853575
A431 Antiproliferative activity assay 72 h IC50 = 0.685 μM 29534926
A431 Antiproliferative activity assay 96 h EC50 = 0.7 μM 28603991
A549 Cytotoxicity assay 72 h IC50 = 0.87 μM 29486953
A431 Antiproliferative activity assay 72 h IC50 = 0.893 μM 27131639
BA/F3 Antiproliferative activity assay 72 h IC50 = 1 μM 26258521
BAF3 Growth inhibition assay 72 h GI50 = 1.2 μM 28282122
NCI-H2122 Function assay 72 h GI50 = 1.2 μM 28282122
A431 Antiproliferative activity assay 72 h IC50 = 1.24 μM 30429956
A431 Antiproliferative activity assay 72 h IC50 = 1.26 μM 29730192
A431 Antiproliferative activity assay 72 h IC50 = 1.604 μM 28033579
A549 Antiproliferative activity assay 96 h EC50 = 1.83 μM 28853575
CHL Growth inhibition assay 72 h GI50 = 2.9 μM 28282122
H1355 Function assay 72 h GI50 = 3 μM 28282122
H1703 Function assay 72 h GI50 = 3.5 μM 28282122
A549 Function assay 72 h GI50 = 3.5 μM 28282122
CHO Growth inhibition assay 72 h GI50 = 4.2 μM 28282122
BAF3 Antiproliferative activity assay 72 h IC50 = 4.61 μM 30429956
BAF3 Antiproliferative activity assay 72 h IC50 = 5.15 μM 30429956
BEAS2B Antiproliferative activity assay 72 h IC50 = 14.9 μM 28716641
NCI-H1975 Function assay 2 h IC50 = 15 μM 25271963
PC9 Function assay 2 h IC50 = 17 μM 25271963
LoVo Function assay 2 h IC50 = 480 μM 25271963
NCI-H1975 Antitumor activity assay Antitumor activity against human NCI-H1975 cells harboring EGFR L858R/T970M double mutant xenografted in SCID mouse assessed as tumor growth inhibition at 2.5 mg/kg/day, po qd for 7 days relative to control 25271963
rat hepatocytes Function assay Intrinsic clearance in rat hepatocytes measured per 10'6 cells 25271963
human hepatocytes Function assay Intrinsic clearance in human hepatocytes measured per 10'6 cells 25271963
NCI-H1975 Antitumor activity assay Antitumor activity against human NCI-H1975 cells harboring EGFR L858R/T970M double mutant xenografted in SCID mouse assessed as tumor growth inhibition at 5 mg/kg/day, po qd for 7 days relative to control 25271963
NCI-H1975 Function assay Selectivity index, ratio of IC50 for EGFR T790M/L858R double mutant expressing human NCI-H1975 cells to IC50 for wild type EGFR expressing human A431 cells 29730192
NCI-H1975 Antitumor activity assay Antitumor activity against EGFR T790M/L858R double mutant expressing human NCI-H1975 cells xenografted in BALB/c athymic nude mouse assessed as tumor growth inhibition at 10 mg/kg, po bid for 21 days 29730192
HCC827 Apoptosis assay Induction of apoptosis in human HCC827 cells harboring EGFR E746-A750 deletion mutant assessed as early apoptotic cells at 3 uM after 24 hrs by Annexin V-FITC/propidium iodide staining based flow cytometry (Rvb = 1.34%) 29466773
HCC827 Apoptosis assay Induction of apoptosis in human HCC827 cells harboring EGFR E746-A750 deletion mutant assessed as late apoptotic cells at 3 uM after 24 hrs by Annexin V-FITC/propidium iodide staining based flow cytometry (Rvb = 27.14%) 29466773
A549 Function assay Selectivity ratio of IC50 for human A549 cells expressing wild-type EGFR/K-Ras mutant to IC50 for human NCI-H1975 cells expressing EGFR L858R/T790M double mutant 29486953
HCC827 Antiproliferative activity assay 72 h Antiproliferative activity against human HCC827 cells at 1 uM after 72 hrs by MTT assay relative to control 29576272
BAF3 Function assay 2 h Inhibition of EGFR T790M/L858R/C797S mutant (unknown origin) expressed in mouse BAF3 cells assessed as reduction in EGF-induced receptor phosphorylation at 1 to 3 uM preincubated for 2 hrs followed by EGF stimulation for 15 mins by Western blot analysis 30429956
BAF3 Function assay 2 h Inhibition of EGFR 19D/T790M/C797S mutant (unknown origin) expressed in mouse BAF3 cells assessed as reduction in EGF-induced receptor phosphorylation at 1 to 3 uM preincubated for 2 hrs followed by EGF stimulation for 15 mins by Western blot analysis 30429956
NCI-H1975 Antitumor activity assay Antitumor activity against human NCI-H1975 cells xenografted in STOCK-Foxn1nu/Nju nude mouse assessed as inhibition of tumor growth at 20 mg/kg/day, po qd for 14 days relative to untreated control 28395219
Caco2 Function assay 2 h Efflux ratio of apparent permeability from basolateral side to apical side over apical side to basolateral side over in human Caco2 cells at 5 uM incubated for 2 hrs 28853575
A431 Function assay Selectivity ratio of EC50 for human A431 cells expressing wild type EGFR to EC50 for human NCI-H1975 cells harboring EGFR-L858R/T790M double mutant 28853575
Caco2 Function assay 2 h Apparent permeability across apical to basolateral side in human Caco2 cells at 5 uM incubated for 2 hrs 28853575
Caco2 Function assay 2 h Apparent permeability across basolateral to apical side in human Caco2 cells at 5 uM incubated for 2 hrs 28853575
NCI-H1975 Function assay 4 h Inhibition of EGFR L858R/T790M mutant in human NCI-H1975 cells assessed as reduction in Akt phosphorylation at Thr308/Ser473 site at 1 uM measured after 4 hrs by Western blot analysis 28282122
HCC827 Function assay 4 h Inhibition of EGFR exon 19 deletion mutant in human HCC827 cells assessed as reduction in Akt phosphorylation at Thr308/Ser473 site at 1 uM measured after 4 hrs by Western blot analysis 28282122
NCI-H1975 Apoptosis assay 48 h Induction of apoptosis in human NCI-H1975 cells harboring EGFR L858R/T790M mutant assessed as caspase3 cleavage at 1 uM after 48 hrs by immunoblotting 28282122
HCC827 Apoptosis assay 48 h Induction of apoptosis in human HCC827 cells harboring EGFR exon 19 deletion mutant assessed as caspase-3 cleavage at 1 uM after 48 hrs by immunoblotting 28282122
PC9 Apoptosis assay 48 h Induction of apoptosis in human PC9 cells harboring EGFR exon 19 deletion mutant assessed as caspase-3 cleavage at 1 uM after 48 hrs by immunoblotting 28282122
H3255 Apoptosis assay 48 h Induction of apoptosis in human H3255 cells harboring EGFR L858R mutant assessed as caspase-3 cleavage at 1 uM after 48 hrs by immunoblotting 28282122
NCI-H1975 Apoptosis assay 48 h Induction of apoptosis in human NCI-H1975 cells harboring EGFR L858R/T790M mutant assessed as PARP cleavage at 1 uM after 48 hrs by immunoblotting 28282122
PC9 Apoptosis assay 48 h Induction of apoptosis in human PC9 cells harboring EGFR exon 19 deletion mutant assessed as PARP cleavage at 1 uM after 48 hrs by immunoblotting 28282122
HCC827 Apoptosis assay 48 h Induction of apoptosis in human HCC827 cells harboring EGFR exon 19 deletion mutant assessed as PARP cleavage at 1 uM after 48 hrs by immunoblotting 28282122
H3255 Apoptosis assay 48 h Induction of apoptosis in human H3255 cells harboring EGFR L858R mutant assessed as PARP cleavage at 1 uM after 48 hrs by immunoblotting 28282122
NCI-H1975 Function assay Selectivity ratio of IC50 for human NCI-H1975 cells harboring EGFR L858R/T790M double mutant to IC50 for human A431 cells harboring wild-type EGFR 28426996
human hepatocytes Function assay Intrinsic clearance in human hepatocytes assessed per million cells 28426996
rat hepatocytes Function assay Intrinsic clearance in rat hepatocytes assessed per million cells 28426996
A549, NCI-H1975 Function assay Selectivity ratio of IC50 for EGF-stimulated wild type EGFR in human A549 cells to IC50 for EGFR L858R/T790M double mutant in human NCI-H1975 cells 26756222
A549, PC9 Function assay Selectivity ratio of IC50 for EGF-stimulated wild type EGFR in human A549 cells to IC50 for EGFR deletion mutant in human PC9 cells 26756222
HCC827 Apoptosis assay Induction of apoptosis in human HCC827 cells harboring EGFR E746 to A750 deletion mutant assessed as early apoptotic cells at 3 uM after 24 hrs by Annexin V-FITC/propidium iodide staining based flow cytometry (Rvb = 1.34 to 1.67%) 29853340
NCI-H1975 Function assay Inhibition of EGFR L858R/T790M double mutant phosphorylation in EGF-stimulated human NCI-H1975 cells at 1 to 100 nM by Western blot method 29906114
A431 Function assay Selectivity ratio, ratio IC50 for human A431 cells overexpressing wild-type EGFR to IC50 for human NCI-H1975 cells expressing EGFR T790M/L858R mutant 27131639
PC9 Antitumor activity assay Antitumor activity against human PC9 cells harboring EGFR exon 19 deletion activating mutant xenografted in SCID mouse assessed as tumor growth inhibition at 10 mg/kg/day, po qd for 7 days relative to control 25271963
A431 Antitumor activity assay Antitumor activity against human A431 cells xenografted in SCID mouse assessed as tumor growth inhibition at 5 mg/kg/day, po qd for 7 days relative to control 25271963
Click to View More Cell Line Experimental Data

Biological Activity

Description Osimertinib (AZD9291) is an oral, irreversible, and mutant-selective EGFR inhibitor with IC50 of 12.92, 11.44 and 493.8 nM for Exon 19 deletion EGFR, L858R/T790M EGFR, and WT EGFR in LoVo cells, respectively. Phase 3.
Features Orally bioavailable mutant-selective EGFR inhibitor that has been tested in Phase III clinical trials for treatment of Non-Small Cell Lung Cancer.
Targets
L858R/T790M EGFR [1]
(LoVo cells)
Exon 19 deletion EGFR [1]
(LoVo cells)
WT EGFR [1]
(LoVo cells)
11.44 nM 12.92 nM 493.8 nM
In vitro
In vitro

AZD9291 shows significantly more potent inhibition of proliferation in mutant EGFR cell lines compared to wild-type in vitro. [2]

Kinase Assay EGFR cellular phosphorylation assay
Cells are seeded (10000 cells/well) in growth medium in Corning black, clear- bottomed 384-well plates and incubated at 37°C with 5% CO2 overnight. Cells are acoustically dosed using an Echo 555, with compounds serially diluted in 100% DMSO. Plates are incubated for a further 2h, then following aspiration of medium, 40μL lx lysis buffer is added to each well. Greiner black high bind 384-well plates are coated with capture antibody and then blocked with 3% BSA. Following removal of block, 15μL of lysate are transferred to the Greiner black high bind 384-well plates and incubated for 2 hours. Following aspiration and washing of the plates with PBS, 20μL, of detection antibody were added and incubated for 2 hours. Following aspiration and washing of the plates with PBS, 20μL of QuantaBlu fluorogenic peroxidase substrate are added and incubated for 1 hour. 20μL QuantaBlu stop solution are added to plates and fluorescence read on an Envision plate reader using Excitation 352nm wavelength and emission 460nm wavelength. The data obtained with each compound is exported into a suitable software package to perform curve fitting analysis. From this data an IC50 value is determined by calculation of the concentration of compound that is required to give a 50% effect.
Cell Research Cell lines PC9 cells
Concentrations 10, 50, & 100 nM
Incubation Time 24 h
Method

Cells were treated with increasing concentrations of osimertinib for 24h.

Experimental Result Images Methods Biomarkers Images PMID
Western blot p-EGFR / p-AKT / p-ERK ABCB1 28416483
Growth inhibition assay Cell viability 31043587
Immunofluorescence Ki67 / γH2AX / p16 29212784
In Vivo
In vivo

AZD9291(5mg/kg p.o.) causes profound regression of tumors across EGFRm+ (PC9) and EGFRm+/T790M (H1975) tumor models with profound inhibition of EGFR phosphorylation and key downstream signaling pathways such as AKT and ERK in vivo. [2]

Animal Research Animal Models Mice bearing PC9 and H1975 xenograft tumors
Dosages ~5 mg/kg
Administration p.o.
NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT05954871 Recruiting
Colorectal Cancer|Non-Small Cell Lung Cancer
Genentech Inc.
January 8 2024 Phase 1
NCT06093945 Completed
Malignant Tumor
Jiangsu HengRui Medicine Co. Ltd.
November 6 2023 Phase 1
NCT06053099 Not yet recruiting
Non Small Cell Lung Cancer|EGFR Activating Mutation|EGFR DEL19|EGFR L858R
Intergroupe Francophone de Cancerologie Thoracique
October 2023 Not Applicable
NCT05401110 Recruiting
Non Small Cell Lung Cancer|Lung Cancer
Karen Reckamp MD MS|Enviro Therapeutics Inc.|Cedars-Sinai Medical Center
September 15 2023 Phase 1
NCT06068049 Recruiting
Non-small Cell Lung Cancer
AstraZeneca
July 28 2023 --
NCT05748093 Not yet recruiting
Non-small Cell Lung Cancer
Maastricht University Medical Center
June 1 2023 Phase 4

Chemical Information & Solubility

Molecular Weight 499.61 Formula

C28 H33 N7 O2

CAS No. 1421373-65-0 SDF Download Osimertinib (AZD9291) SDF
Smiles CN1C=C(C2=CC=CC=C21)C3=NC(=NC=C3)NC4=C(C=C(C(=C4)NC(=O)C=C)N(C)CCN(C)C)OC
Storage (From the date of receipt)

In vitro
Batch:

DMSO : 99 mg/mL ( (198.15 mM); Moisture-absorbing DMSO reduces solubility. Please use fresh DMSO.)

Ethanol : 99 mg/mL

Water : Insoluble


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In vivo
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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.

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Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Frequently Asked Questions

Question 1:
Can this formulation be used in mice? What are reconstitution instructions for in vivo with mice?

Answer:
Osimertinib can be used for animal study. The vehicle we suggest is: 5%DMSO+40%PEG300+5%Tween 80+50%ddH2O.

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