Erlotinib (OSI-774)

Catalog No.S7786 Synonyms: CP358774, NSC 718781, Tarceva

For research use only.

Erlotinib (OSI-774, CP358774, NSC 718781, Tarceva) is an EGFR inhibitor with IC50 of 2 nM, >1000-fold more sensitive for EGFR than human c-Src or v-Abl. Erlotinib induces autophagy.

Erlotinib (OSI-774) Chemical Structure

CAS No. 183321-74-6

Selleck's Erlotinib (OSI-774) has been cited by 510 publications

Purity & Quality Control

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Biological Activity

Description Erlotinib (OSI-774, CP358774, NSC 718781, Tarceva) is an EGFR inhibitor with IC50 of 2 nM, >1000-fold more sensitive for EGFR than human c-Src or v-Abl. Erlotinib induces autophagy.
Targets
EGFR [1]
(Cell-free assay)
2 nM
In vitro

Erlotinib HCl potently inhibits EGFR activation in intact cells including HNS human head and neck tumor cells (IC50 20nM), DiFi human colon cancer cells and MDA MB-468 human breast cancer cells. Erlotinib HCl (1 μM) induces apoptosis in DiFi human colon cancer cells. [1] Erlotinib inhibits growth of a panel of NSCLC cell lines including A549, H322, H3255, H358 H661, H1650, H1975, H1299, H596 with IC50 ranging from 29 nM to >20 μM. [2] Erlotinib HCl(2 μM) significantly inhibits growth of AsPC-1 and BxPC-3 pancreatic cells. [3] The effects of Erlotinib HCl in combination with gemcitabine are considered additive in KRAS-mutated pancreatic cancer cells. Ten micromolar of Erlotinib HCl inhibits EGFR phospho-rylation at the Y845 (Src-dependent phosphorylation) and Y1068 (auto-phosphorylation) sites. [4] Combination with Erlotinib HCl could down-modulate rapamycin-stimulated Akt activity and produces a synergistic effect on cell growth inhibition. [5]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
human SKBR3 cells MmTGSpVv[3Srb36gZZN{[Xl? M3zSTGlvcGmkaYTpc44hd2ZiSFXSNkBqdiCqdX3hckBUU0KUMzDj[YxteyCkeTDIWHJHKGG|c3H5MEBKSzVyPUGuPFkh|ryPLh?= M4Hod|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF7OEG1OFEzLz5zOUixOVQyOjxxYU6=
human A431 cells MofHSpVv[3Srb36gZZN{[Xl? M{\5OWlvcGmkaYTpc44hd2ZiRVfGVkBqdiCqdX3hckBCPDNzIHPlcIx{KGK7IFjUVmYh[XO|YYmsJGlEPTB;MD60NkDPxE1w MYq8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yQThzNUSxNkc,OTl6MUW0NVI9N2F-
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T24 NIH MmfvSpVv[3Srb36gZZN{[Xl? M4q0SGlvcGmkaYTpc44hd2ZibHnnZY5lKHO2aX31cIF1\WRiZYLiRlIh[XW2b4Doc5NxcG:{eXzheIlwdiCrbjDUNlQhVkmKIHPlcIx{NCCLQ{WwJF0hOC5{OUmg{txONg>? MmSyQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTZ2OECyPFQoRjF4NEiwNlg1RC:jPh?=
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erythroleukemia cells Mnr2S5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NU\OVG5zOjRiaILz Mn:0S5Jwf3SqIHnubIljcXSrb36gc4YhdW:3c3Wg[ZJ6fGi{b3zleYtmdWmjIHPlcIx{KGGodHXyJFI1KGi{czygTWM2OCB;IEKg{txONg>? MnXYQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTdzN{i3NlIoRjF5MUe4O|IzRC:jPh?=
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HepG2 M{jGTmFvfGmycn;sbYZmemG2aY\lJIF{e2G7 MXu0PEBpenN? M1\He2FvfGmycn;sbYZmemG2aY\lJIFkfGm4aYT5JIFo[Wmwc4SgbJVu[W5iSHXwS|Ih[2WubIOgZYZ1\XJiNEigbJJ{KGK7IF3UWEBie3OjeTygTWM2OCB;IECuNVIh|ryPLh?= MlzJQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN7NkK2OlAoRjJ|OU[yOlYxRC:jPh?=
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NCI-H1975 MYLDfZRwfG:6aXPpeJkh[XO|YYm= MX[3NkBpenN? NH\ZOGFEgXSxdH;4bYNqfHliYXfhbY5{fCCqdX3hckBPS0lvSEG5O|Uh[2WubIOg[ZhxemW|c3nu[{BGT0[UIFy4OVhTN1R5OUDNJI12fGGwdDDh[pRmeiB5MjDodpMh[nliTWTUJIF{e2G7LDDJR|UxKD1iNT61NUDPxE1w M3;B[VxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NEGxNVI{Lz5{NESxNVEzOzxxYU6=
A431 NHrkZ3BCdnSrcILvcIln\XKjdHn2[UBie3OjeR?= M3PwR|czKGi{cx?= NEXRcpZCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIFG0N|Eh[2WubIOgZZN{\XO|ZXSgZZMh\3Kxd4ToJIlvcGmkaYTpc44h[W[2ZYKgO|IhcHK|IHL5JHNTSiCjc4PhfUwhUUN3MDC9JFAvPTZizszNMi=> NH3rfVI9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEW2OVk3QSd-MkS1OlU6Pjl:L3G+
NCI-H1975 MYjBcpRqeHKxbHnm[ZJifGm4ZTDhd5NigQ>? MlPUO|IhcHK| NVfmVGlTSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[WejaX7zeEBo\W[rdHnubYIuemW|aYP0ZY51KGi3bXHuJG5EUS2KMUm3OUBk\WyuczDhd5Nme3OnZDDhd{Boem:5dHigbY5pcWKrdHnvckBi\nSncjC3NkBpenNiYomgV3JDKGG|c3H5MEBKSzVyIE2gO{4xPiEQvF2u MkDVQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR3NkW5OlkoRjJ2NU[1PVY6RC:jPh?=
Sf9 NXe4WmpzTnWwY4Tpc44h[XO|YYm= NGr6bmIyKGi{ M2r4SmlvcGmkaYTpc44hd2ZiYYX0c5Bpd3OyaH;yfYxifGmxbjDv[kBz\WOxbXLpcoFvfCCHR1\SJIN6fG:ybHHzcYlkKGSxbXHpckApPjR3IITvJFEyQDZrIDj1cotvd3ewIH;ybYdqdiliZYjwdoV{e2WmIHnuJHNnQSClZXzsd{Bi\nSncjCxJIhzKGK7IFTFUGZKSS:2aX3lMZJme2:udnXkJIZtfW:{b33leJJq[yCjbnHsfZNqeyxiSVO1NEA:KDBwMEOyJO69VS5? M2mwc|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NkC3PVk5Lz5{NE[wO|k6QDxxYU6=
HepG2 MXLBcpRqeHKxbHnm[ZJifGm4ZTDhd5NigQ>? NHXJO5c1QCCqcoO= MmXZRY51cXC{b3zp[oVz[XSrdnWgZYN1cX[rdImgZYdicW6|dDDoeY1idiCKZYDHNkBk\WyuczDh[pRmeiB2ODDodpMh[nliTWTUJIF{e2G7LDDJR|UxKD1iMD6xNkDPxE1w NF;jUYY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NE[wO|k6QCd-MkS2NFc6QTh:L3G+
A549 NIjTV5hCdnSrcILvcIln\XKjdHn2[UBie3OjeR?= NYLWXppwPDhiaILz NIKwZZpCdnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIFG1OFkh[2WubIOgZYZ1\XJiNEigbJJ{KGK7IF3UWEBie3OjeTygTWM2OCB;IECuNVMh|ryPLh?= MkDQQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR4MEe5PVgoRjJ2NkC3PVk5RC:jPh?=
Sf9 M2rINGZ2dmO2aX;uJIF{e2G7 Mmm5NUBpeg>? M4jBZ2lvcGmkaYTpc44hd2ZiYYX0c5Bpd3OyaH;yfYxifGmxbjDv[kBGT0[UIHP5eI9xdGG|bXnjJIRwdWGrbjCoZY1qdm9iYXPp[JMhPjR3IITvJFEyQDZrIDj1cotvd3ewIH;ybYdqdiliZYjwdoV{e2WmIHnuJHNnQSClZXzsd{Bi\nSncjCxJIhzKGK7IITpcYUuemW|b3z2[YQh\my3b4LvcYV1enluIFnDOVAhRSByLkCzNkDPxE1w M2jjVlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NkOwOFEzLz5{NE[zNFQyOjxxYU6=
HepG2 MkTlRY51cXC{b3zp[oVz[XSrdnWgZZN{[Xl? MoXwOFghcHK| MmS0RY51cXC{b3zp[oVz[XSrdnWgZYN1cX[rdImgZYdicW6|dDDoeY1idiCKZYDHNkBk\WyuczDh[pRmeiB2ODDodpMh[nliTWTUJIF{e2G7LDDJR|UxKD1iMD6xNkDPxE1w NYq5bG1ERGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS2N|A1OTJpPkK0OlMxPDF{PD;hQi=>
A549 M1jnSmFvfGmycn;sbYZmemG2aY\lJIF{e2G7 NX;Rdnp7PDhiaILz M33UXGFvfGmycn;sbYZmemG2aY\lJIFkfGm4aYT5JIFo[Wmwc4SgbJVu[W5iQUW0PUBk\WyuczDh[pRmeiB2ODDodpMh[nliTWTUJIF{e2G7LDDJR|UxKD1iMD6xN{DPxE1w M2\WO|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NkOwOFEzLz5{NE[zNFQyOjxxYU6=
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SCC-4 NUjEc5RDT3Kxd4ToJGlvcGmkaYTpc44h[XO|YYm= NHzxW|VKdmirYnn0bY9vKG:oIHj1cYFvKFOFQz20JINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUCgQUA1QS53NkCyJO69VS5? MUC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:5d4eu[YJqNmGlLoXrM4Np\W2kbD;jc41xd3WwZG;y[ZBwenShY3Hy[E9EUEWPQly1OVMwLz6VQV7HSXI9N2F-
RKO MoDRS5Jwf3SqIFnubIljcXSrb36gZZN{[Xl? NWrQeIltUW6qaXLpeIlwdiCxZjDoeY1idiCUS1:gZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3MDC9JFQ6Njh|NU[g{txONg>? NILlZ2Q9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;3e5cv\WKrLnHjMpVsN2OqZX3icE9kd22yb4Xu[H9z\XCxcoTfZ4Fz\C:FSFXNRmw2PTNxJ{7TRW5ITVJ:L3G+
Assay
Methods Test Index PMID
Western blot Cleaved PARP / PARP ; P-Raf-1 / t-Raf-1 / p-MEK / t-MEK / p-ERK / t-ERK / p-PI3K / t-PI3K / p-AKT / AKT ; p-STAT3 / STAT3 / PTPMeg2 / p-EGFR / EGFR / pJAK2 / JAK2 / Bcl2 / Bcl-xl / Survivin 27508092 24019973
Immunofluorescence β-catenin ; E-cadherin / Vimentin 25209444 19825949
Growth inhibition assay Cell viability ; IC50 30377412 17671085
In vivo At doses of 100 mg/kg, Erlotinib HCl completely prevents EGF-induced autophosphorylation of EGFR in human HN5 tumors growing as xenografts in athymic mice and of the hepatic EGFR of the treated mice. [1] Erlotinib HCl (100 mg/Kg) inhibits H460a and A549 tumor models with 71 and 93% inhibition rate. [5]

Protocol (from reference)

Kinase Assay:

[1]

  • Kinase assays:

    96-well plates are coated by incubation overnight at 37 °C with 100 μL per well of 0.25 mg/mL PGT in PBS. Excess PGT is removed by aspiration, and the plate is washed 3 times with washing buffer (0.1% Tween 20 in PBS). The kinase reaction is performed in 50 μL of 50 mM HEPES (pH 7.3), containing 125 mM sodium chloride, 24 mM magnesium chloride, 0.1 mM sodium orthovanadate, 20 μM ATP, 1.6 μg/mL EGF, and 15 ng of EGFR, affinity purified from A431 cell membranes. Erlotinib HCl in DMSO is added to give a final DMSO concentration of 2.5%. Phosphorylation is initiated by addition of ATP and proceeded for 8 minutes at room temperature, with constant shaking. The kinase reaction is terminated by aspiration of the reaction mixture and is washed 4 times with washing buffer. Phosphorylated PGT is measured by 25 minutes of incubation with 50 μL per well HRP-conjugated PY54 antiphosphotyrosine antibody, diluted to 0.2 μg/mL in blocking buffer (3% BSA and 0.05% Tween 20 in PBS). Antibody is removed by aspiration, and the plate is washed 4 times with washing buffer. The colorimetric signal is developed by addition of TMB Microwell Peroxidase Substrate, 50μL per well, and stopped by the addition of 0.09 M sulfuric acid, 50 μL per well. Phosphotyrosine is estimated by measurement of absorbance at 450 nm. The signal for controls is typically 0.6-1.2 absorbance units, with essentially no back ground in wells without AlP, EGFR, or PGT and is proportional to the time of incubation for 10 minutes.

Cell Research:

[2]

  • Cell lines: A549, H322, H3255, H358 H661, H1650, H1975, H1299, H596 cells
  • Concentrations: 30 nM-20 μM
  • Incubation Time: 72 hours
  • Method:

    Exponentially growing cells are seeded in 96-well plastic plates and exposed to serial dilutions of erlotinib, pemetrexed, or the combination at a constant concentration ratio of 4:1 in triplicates for 72 h. Cell viability is assayed by cell count and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Growth inhibition is expressed as the percentage of surviving cells in drug-treated versus PBS-treated control cells (which is considered as 100% viability). The IC50 value is the concentration resulting in 50% cell growth inhibition by a 72-h exposure to drug(s) compared with untreated control cells and is calculated by the CalcuSyn software.

Animal Research:

[6]

  • Animal Models: Male 5-week-old BALB-nu/nu mice with HPAC cells
  • Dosages: 50 mg/kg
  • Administration: Oral administration

Solubility (25°C)

In vitro

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
5% DMSO+30% PEG 300+5% Tween 80+ddH2O
For best results, use promptly after mixing.

3mg/mL

Chemical Information

Molecular Weight 393.44
Formula

C22H23N3O4

CAS No. 183321-74-6
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles COCCOC1=C(C=C2C(=C1)C(=NC=N2)NC3=CC=CC(=C3)C#C)OCCOC

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Calculation results:

Working concentration: mg/ml;

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.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Clinical Trial Information

NCT Number Recruitment Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT04172779 Not yet recruiting Drug: Erlotinib Hydrochloride Cirrhosis Liver University of Texas Southwestern Medical Center July 2022 Phase 2
NCT03460678 Terminated Drug: Pemetrexed|Drug: Erlotinib Carcinoma Non-Small-Cell Lung Hikma Pharmaceuticals LLC February 28 2018 Phase 4
NCT02942095 Active not recruiting Drug: Ixazomib|Drug: Erlotinib Solid Tumors M.D. Anderson Cancer Center|Millennium: The Takeda Oncology Company March 6 2017 Phase 1
NCT02991651 Recruiting Drug: IRX4204|Drug: erlotinib Lung Cancer Nonsmall Cell Io Therapeutics|Dartmouth College May 2016 Phase 1

(data from https://clinicaltrials.gov, updated on 2022-01-17)

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 you please give me some advice for which solvent we should use to dissolve it for animal studies?

Answer:
For in vivo application, we recommend to use 5% DMSO+45% PEG 300+ddH2O up to 6mg/ml.

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