Rapamycin (AY-22989)

For research use only.

Licensed by Pfizer Catalog No.S1039 Synonyms: Rapamune, Sirolimus, NSC-2260804

1350 publications

Rapamycin (AY-22989) Chemical Structure

CAS No. 53123-88-9

Rapamycin (AY-22989, Rapamune, Sirolimus, NSC-2260804) is a specific mTOR inhibitor with IC50 of ~0.1 nM HEK293 cells.

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Selleck's Rapamycin (AY-22989) has been cited by 1350 publications

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Biological Activity

Description Rapamycin (AY-22989, Rapamune, Sirolimus, NSC-2260804) is a specific mTOR inhibitor with IC50 of ~0.1 nM HEK293 cells.
Targets
mTOR [1]
(HEK293 cells)
~0.1 nM
In vitro

Rapamycin inhibits endogenous mTOR activity in HEK293 cells with IC50 of ~0.1 nM, more potently than iRap and AP21967 with IC50 of ~5 nM and ~10 nM, respectively. [1] In Saccharomyces cerevisiae, Rapamycin treatment induces a severe G1/S cell cycle arrest and inhibition of translation initiation to levels below 20% of control. [2] Rapamycin significantly inhibits the cell viability of T98G and U87-MG in a dose-dependent manner with IC50 of 2 nM and 1 μM, respectively, while displaying little activity against U373-MG cells with IC50 of >25 μM despite the similar extent of the inhibition of mTOR signaling. Rapamycin (100 nM) induces G1 arrest and autophagy but not apoptosis in Rapamycin-sensitive U87-MG and T98G cells by inhibiting the function of mTOR. [3]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
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HEK293 M3vyNWZ2dmO2aX;uJGF{e2G7 MX6xNFAhdk1? Mo[5PEBp M1vnU2ROW09? NETHTmpKdmirYnn0d{BVWEFvaX7keYNm\CCmZXfyZYRifGmxbjDv[kBR\GOmNDD3bZRpKEWFNUCgc4YhPTBibl2= MkLaQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjF3M{mzNFEoRjJzNUO5N|AyRC:jPh?=
PC3 NG\tcZpIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NYnaPJdQOS53IN88US=> NX\2XYJlOSCq NEDtcWVFVVOR NXjvVXdzUW6mdXPld{BidnSrcILvcIln\XKjdHn2[UBi[3Srdnn0fUBi\2GrboP0JIh2dWGwIGDDN{Bk\WyuczD3bZRpKEmFNUCgc4YhRDFyIH7N MV[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOTl5OE[4N{c,OjF7N{i2PFM9N2F-
PC3 MXnLbY5ie2ViQYPzZZk> NETQN5YyODBibl2= M13LRVEhcA>? NVP3U2RDTE2VTx?= NVjrWIlGTG:nczDuc5QhcW6qaXLpeEBuXE:ULX3l[IlifGWmIFHreEBxcG:|cHjvdplt[XSrb36= NGXMW5A9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MUm3PFY5Oyd-MkG5O|g3QDN:L3G+
PC3 NEXyNFlMcW6jc3WgRZN{[Xl? M1rOSVExOCCwTR?= M3q2bVEhcA>? MXzEUXNQ NYPPU|doWG:2ZX70cJkhcW6qaXLpeJMhdVSRUj3t[YRq[XSnZDDTOkBxcG:|cHjvdplt[XSrb36ge4l1cCCLQ{WwJI9nKDxzMDDuUU4> M2TCfVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJzOUe4Olg{Lz5{MUm3PFY5OzxxYU6=
HT-29 NEWzNHZEgXSxdH;4bYMhSXO|YYm= NVm5UWJVOTBibl2= NGfzOVc4OiCq MofwSG1UVw>? NXXw[HJPWG:2ZX70bYF1\XNiNT3mcJVwem:3cnHjbYwucW6mdXPl[EBkgXSxdH;4bYNqfHl? NVfw[HpuRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS5NFA5PzNpPkK0PVAxQDd|PD;hQi=>
HT-29 Mn7DR5l1d3SxeHnjJGF{e2G7 MV[xNEBvVQ>? NH60XXY4OiCq M3zZR2ROW09? M4fXSXBwfGWwdHnheIV{KGSrZ3n0c5hqdi2rbnT1Z4VlKGO7dH;0c5hq[2m2eR?= M3nKW|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2OUCwPFc{Lz5{NEmwNFg4OzxxYU6=
HT-29 MmDLR5l1d3SxeHnjJGF{e2G7 MVuxNEBvVQ>? NWnEcW1MPzJiaB?= NWDtS5F{TE2VTx?= Mkm0VI91\W62aXH0[ZMh[2GvcITveIhm[2mwLXnu[JVk\WRiY4n0c5RwgGmlaYT5 MVu8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDlyMEi3N{c,OjR7MEC4O|M9N2F-
HEK293 NHP1R5hHfW6ldHnvckBCe3OjeR?= NVz6cIkyUUN3MDC9JFAvODByMTFOwG0> MlvuQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd|NUC5OVMoRjF5M{WwPVU{RC:jPh?=
HEK293T, PBMCs NITsRpJCdnSrdnnyZYwh[WO2aY\peJkh[XO|YYm= MWS2JIRigXN? M1fmemVEPTBiPTCwMlAxODFizszN MnHzQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTd2OEW1NFEoRjF5NEi1OVAyRC:jPh?=
HEK293T, PBMCs NHPZRWxCdnSrdnnyZYwh[WO2aY\peJkh[XO|YYm= MUK2JIRigXN? MlXhSWM2OCB;IECuNFAyOyEQvF2= NFrJXXY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zN{S4OVUxOSd-MUe0PFU2ODF:L3G+
cells from thymus of normal BALB/c mice MXXGeY5kfGmxbjDBd5NigQ>? MoPCTWM2OCB;IECuNFA{KM7:TR?= NGj1[oo9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zMECyNVk1QCd-MUCwNlE6PDh:L3G+
PC3 NVPIe5htdVSRUjDpcohq[mm2aX;uJIF{e2G7 M3fiZmlEPTBiPTCwMlAyKM7:TR?= MlXXQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjF7N{i2PFMoRjJzOUe4Olg{RC:jPh?=
PC3 NXK0fpJDSW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[XO|YYm= Mn3DTWM2OCB;IECuNFEh|ryP NGnnVpg9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MUm3PFY5Oyd-MkG5O|g3QDN:L3G+
HEK293 NYXNVlVOTnWwY4Tpc44hSXO|YYm= NX;RSmxTQCCq M1G3NmVEPTBiPTCwMlA2KM7:TR?= M4SwZVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJzNUO5N|AyLz5{MUWzPVMxOTxxYU6=
HeLa NUWyVWd7S3m2b4TvfIlkcXS7IHHzd4F6 MYXJR|UxKD1iMD63O{DPxE1? NFfPOm89[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9|MECxOVA4OCd-M{CwNVUxPzB:L3G+
HEK293 NYT1cXBHTnWwY4Tpc44hSXO|YYm= NEPVVZVKSzVyIE2gNU4yKM7:TR?= NYHYdHBiRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMUS1N|A6ODdpPkG0OVMxQTB5PD;hQi=>
HCT-8 NWnpXFVVTnWwY4Tpc44hSXO|YYm= MVfJR|UxKD1iMT6yOUDPxE1? NI\oe2E9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zMkKzOVI3PSd-MUKyN|UzPjV:L3G+
T47D MVTHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MWe3NkBp MlfCTWM2OCB;IEGuOVc3KM7:TR?= M3ew[lxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ7M{C4PFk2Lz5{OUOwPFg6PTxxYU6=
SiHa M2fpWGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MWe3NkBp NFHV[HVKSzVyIE2gNU44PTZizszN NUj4NFlRRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkmzNFg5QTVpPkK5N|A5QDl3PD;hQi=>
Lewis rat lymph node NV\Jc5N3SW62aYDyc4xq\mW{YYTpeoUh[WO2aY\peJkh[XO|YYm= MVXJR|UxKD1iMj62JO69VQ>? MV68ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8yPjF6NUi2OUc,OTZzOEW4OlU9N2F-
MCF7 M3LBeWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MmXJO|IhcA>? NUjoXYhWUUN3MDC9JFQvQThizszN M{fOOVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ7M{C4PFk2Lz5{OUOwPFg6PTxxYU6=
HepG2 (DPX-2) M2jObWZ2dmO2aX;uJGF{e2G7 NEmyS5UzPCCq M1TwOGVEPTBiPTCxNEDPxE1? MlPiQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjB7Nk[wOFMoRjJyOU[2NFQ{RC:jPh?=
Vero E6 MoDKRY51cX[rcnHsJIFkfGm4aYT5JIF{e2G7 Mk\BTWM2OCB;IEK2MlMxOjdizszN MXq8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8{OjN3M{i1PUc,OzJ|NUO4OVk9N2F-
PC3 M3rn[mN6fG:2b4jpZ4l1gSCjc4PhfS=> MUfFR|UxKD1iNEOuNUDPxE1? M{XETlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ6N{e0OFI3Lz5{OEe3OFQzPjxxYU6=

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
p-mTOR(S2448)/mTOR; 

PubMed: 23991038     


MCF-7-vector and MCF-7-ZNF703 cells were treated with DMSO, 0.1 µM rapamycin, or 1 µM rapamycin for 24 hours before cell lysis. Western blot analysis of lysates using the indicated antibodies. A GAPDH antibody was used as a loading control.

23991038
Growth inhibition assay
Cell proliferation; 

PubMed: 30393233     


(A) Effect of Rapamycin treatment on SK-N-SH cell proliferation. (B) Effect of Rapamycin treatment on SH-SY5Y cell proliferation. Rapamycin, except treatment of 10 µM for 12 h, can obviously inhibited proliferation of NB cells compared with corresponding control group (P<0.05). (*) indicates statistically significant difference with P<0.05.

30393233
Histomorphology
Haematoxylin & Eosin; 

PubMed: 28418837     


Representative pictures of H&E-stained U87MG cells. The fusiform cell body observed in control cells A. disappears in cells exposed to increasing doses of rapamycin for 24 h B.-E. Rapamycin produces a dose-dependent increase in the diameter of the cell body which develops a pyramidal shape. 1 nM (B); 10 nM (C); 100 nM (D); 1 μM (E) of rapamycin exposure.

28418837
Immunofluorescence
NeuN; 

PubMed: 28418837     


Immune-fluorescence of U87MG cells treated with vehicle A.-D. and rapamycin at the dose of 10 nM E.-H.; 100 nM I.-L.; 1 μM M.-P. In the first line cells were stained for the late post-mitotic neuronal marker NeuN. In the second line cells were stained for the nuclear dye DAPI. In the third line the merging between NeuN (green) and DAPI (blue) fluorescence is shown. In the fourth line a high magnification of the squared insert of line three is shown.

p62/Beclin; 

PubMed: 28819214     


(C,D) Rapamycin treatment reduced the aggregate level of p62 and Beclin 1 in cardiomyocytes. NRVCs were transduced with Ad-Nef or Ad-null for 48 hours and cells were fixed with 4% PFA. Fixed cells were stained with p62 and Beclin 1 antibody. 

28418837 28819214
ELISA
Type III collagen/Fibronectin; 

PubMed: 23364979     


(A) Rapamycin inhibits TGF-β1-induced type III collagen level in the culture medium of human lung fibroblast, evidenced by ELISA. Type III collagen level after 24-h incubation with medium alone (control), 0.01, 0.1, 1.0, and 10 ng/mL rapamycin and 10 ng/mL TGF-β1. No significant difference was found between different concentration groups of rapamycin. (B) Rapamycin inhibits TGF-β1-induced fibronectin level in the culture medium of human lung fibroblast, shown by ELISA. Fibronectin level after 24-h incubation with medium alone (control), 0.01, 0.1, 1.0, and 10 ng/mL rapamycin and 10 ng/mL TGF-β1. Values are mean of three independent experiments. No significant difference was found between different concentration groups of rapamycin. *p<0.05 vs. control; †p<0.05 vs. TGF-β1 group. TGF-β1, transforming growth factor β1.

23364979
In vivo Treatment with Rapamycin in vivo specifically blocks targets known to be downstream of mTOR such as the phosphorylation and activation of p70S6K and the release of inhibition of eIF4E by PHAS-1/4E-BP1, leading to complete blockage of the hypertrophic increases in plantaris muscle weight and fibre size. [4] Short-term Rapamycin treatment, even at the lowest dose of 0.16 mg/kg, produces profound inhibition of p70S6K activity, which correlates with increased tumor cell death and necrosis of the Eker renal tumors. [5] Rapamycin inhibits metastatic tumor growth and angiogenesis in CT-26 xenograft models by reducing the production of VEGF and blockage of VEGF-induced endothelial cell signaling. [6] Rapamycin treatment at 4 mg/kg/day significantly reduces tumor growth of C6 xenografts, and tumor vascular permeability. [7]

Protocol

Kinase Assay:

[1]

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Immunoblotting for the mTOR kinase assay:

HEK293 cells are plated at 2-2.5×105 cells/well of a 12-well plate and serum-starved for 24 hours in DMEM. Cells are treated with increasing concentrations of Rapamycin (0.05-50 nM) for 15 minutes at 37 °C. Serum is added to a final concentration of 20% for 30 minutes at 37 °C. Cells are lysed, and cell lysates are separated by SDS-PAGE. Resolved proteins are transferred to a polyvinylidene difluoride membrane and immunoblotted with a phosphospecific primary antibody against Thr-389 of p70 S6 kinase. Data are analyzed using ImageQuant and KaleidaGr
Cell Research:

[3]

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  • Cell lines: U87-MG, T98G, and U373-MG
  • Concentrations: Dissolved in DMSO, final concentrations ~25 μM
  • Incubation Time: 72 hours
  • Method:

    Cells are exposed to various concentrations of Rapamycin for 72 hours. For the assessment of cell viability, cells are collected by trypsinization, stained with trypan blue, and the viable cells in each well are counted. For the determination of cell cycle, cells are trypsinized, fixed with 70% ethanol, and stained with propidium iodide using a flow cytometry reagent set. Samples are analyzed for DNA content using a FACScan flow cytometer and CellQuest software. For apoptosis detection, cells are stained with the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) technique using an ApopTag apoptosis detection kit. To detect the development of acidic vesicular organelles (AVO), cells are stained with acridine orange (1 μg/mL) for 15 minutes, and examined under a fluorescence microscope. To quantify the development of AVOs, cells are stained with acridine orange (1 μg/mL) for 15 minutes, removed from the plate with trypsin-EDTA, and analyzed using the FACScan flow cytometer and CellQuest software. To analyze the autophagic process, cells are incubated for 10 minutes with 0.05 mM monodansylcadaverine at 37 °C and are then observed under a fluorescence microscope.


    (Only for Reference)
Animal Research:

[7]

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  • Animal Models: Athymic Nu/Nu mice inoculated subcutaneously with VEGF-A-expressing C6 rat glioma cells
  • Dosages: ~4 mg/kg/day
  • Administration: Injection i.p.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 20 mg/mL (21.87 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
2% DMSO+30% PEG 300+5% Tween 80+ddH2O
For best results, use promptly after mixing.
5mg/mL

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 914.18
Formula

C51H79NO13

CAS No. 53123-88-9
Storage powder
in solvent
Synonyms Rapamune, Sirolimus, NSC-2260804
Smiles CC1CCC2CC(C(=CC=CC=CC(CC(C(=O)C(C(C(=CC(C(=O)CC(OC(=O)C3CCCCN3C(=O)C(=O)C1(O2)O)C(C)CC4CCC(C(C4)OC)O)C)C)O)OC)C)C)C)OC

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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Step 2: Enter the in vivo formulation ()
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Bio Calculators

Molarity Calculator

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Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

Mass (mg) = Concentration (mM) × Volume (mL) × Molecular Weight (g/mol)

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Dilution Calculator

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Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

Concentration (start) x Volume (start) = Concentration (final) x Volume (final)

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Clinical Trial Information

NCT Number Recruitment interventions Conditions Sponsor/Collaborators Start Date Phases
NCT04629495 Not yet recruiting Drug: Rapamycin|Other: Placebo Mild Cognitive Impairment|Alzheimer Disease The University of Texas Health Science Center at San Antonio April 2021 Phase 2
NCT04172922 Not yet recruiting Drug: Topical Sirolimus Vascular Anomaly Nemours Children''s Clinic December 1 2019 Phase 1
NCT03298958 Withdrawn Drug: Rapamycin|Drug: Placebo Oral Tablet Bladder Cancer The University of Texas Health Science Center at San Antonio November 1 2018 Phase 3
NCT03243019 Recruiting Drug: rapamycin|Device: MRI|Biological: Rapamycin dosage Lymphatic Malformation|Pediatric University Hospital Lille|Ministry of Health France June 25 2018 Phase 2
NCT04598204 Recruiting Drug: Treatment with oral rapmycin Vascular Anomalies Zhujiang Hospital December 1 2017 Phase 2|Phase 3
NCT03359538 Active not recruiting Drug: Rapamycin|Drug: Placebo Oral Tablet Amyotrophic Lateral Sclerosis Azienda Ospedaliero-Universitaria di Modena|University of Modena and Reggio Emilia|Azienda Ospedaliero Universitaria Maggiore della Carita|IRCCS Azienda Ospedaliera Universitaria San Martino - IST Istituto Nazionale per la Ricerca sul Cancro Genoa Italy|University of Turin Italy|Fondazione I.R.C.C.S. Istituto Neurologico Carlo Besta|Azienda Ospedaliera Niguarda Cà Granda|Fondazione Salvatore Maugeri|University of Padova September 19 2017 Phase 2

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.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID