Rapamycin (Sirolimus; AY22989)

Alias: AY 22989; AY22989; AY-22989; NSC-2260804; RAPA; RAP; RPM; SLM; AY 22989; SILA 9268A; WY090217; WY-090217; WY 090217; C07909; D00753; Rapamune
Cat No.:V0174 Purity: ≥98%
Rapamycin (also known as Sirolimus; AY-22989), a natural macrocyclic lactone isolated from the bacterium Streptomyces hygroscopicus, is a specific and potent mTOR inhibitor with IC50 of ~0.1 nM in HEK293 cells.
Rapamycin (Sirolimus; AY22989) Chemical Structure CAS No.: 53123-88-9
Product category: mTOR
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
5mg
50mg
100mg
250mg
500mg
1g
2g
Other Sizes

Other Forms of Rapamycin (Sirolimus; AY22989):

  • 28-Epirapamycin
  • Rapamycin analog-2
  • Seco Rapamycin (Secorapamycin; Secorapamycin A)
  • Rapamycin-13C,d3 (rapamycin; sirolimus-13C,d3; Sirolimus-13C,d3; AY-22989-13C,d3)
  • Seco Rapamycin ethyl ester
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Purity & Quality Control Documentation

Purity: ≥98%

Purity: =99.046%

Purity: ≥98%

Product Description

Rapamycin (also known as Sirolimus; AY-22989), a natural macrocyclic lactone isolated from the bacterium Streptomyces hygroscopicus, is a specific and potent mTOR inhibitor with IC50 of ~0.1 nM in HEK293 cells. Although rapamycin was initially created as an antifungal antibiotic, it also showed signs of immunosuppressive activity, and it is now used for this reason to prevent transplant rejection. Additionally, it shows activity against a number of transplantable tumors and is only marginally to completely inactive against leukemias. Rapamycin suppresses the immune system by preventing T cells from activating and proliferating. The rapamycin-FKBP12 complex, which is formed when rapamycin binds to FK-binding protein 12 (FKBP12), controls an enzyme that is crucial to the progression of the cell cycle.

Biological Activity I Assay Protocols (From Reference)
Targets
mTOR (IC50 = 0.1 nM); Microbial Metabolite; Autophagy; Human Endogenous Metabolite
ln 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] Rapamycin treatment causes a severe G1/S cell cycle arrest in Saccharomyces cerevisiae and inhibits translation initiation to levels below 20% of control. [2] Rapamycin exhibits little activity against U373-MG cells with an IC50 of >25 M despite having a similar impact on the inhibition of mTOR signaling. Rapamycin significantly reduces the cell viability of T98G and U87-MG in a dose-dependent manner. By inhibiting the activity of mTOR, rapamycin (100 nM) causes G1 arrest and autophagy but not apoptosis in Rapamycin-sensitive U87-MG and T98G cells. [3]
ln 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, results in profound inhibition of p70S6K activity, which is correlated with an increase in tumor cell death and necrosis of the Eker renal tumors. [5] By lowering VEGF production and preventing VEGF-induced endothelial cell signaling, rapamycin inhibits angiogenesis and metastatic tumor growth in CT-26 xenograft models. [6] Rapamycin treatment at 4 mg/kg/day significantly reduces tumor vascular permeability and tumor growth in C6 xenografts. [7]
Enzyme 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. Rapamycin (0.05–50 nM) is administered to cells for 15 minutes at 37 °C in escalating concentrations. 30 minutes at 37 °C are spent adding serum at a final concentration of 20%. Cell lysates are separated by SDS-PAGE after being lysed. Proteins that have been resolved are transferred to a polyvinylidene difluoride membrane and immunoblotted using a primary antibody that is phosphospecific for the Thr-389 of p70 S6 kinase. using ImageQuant and KaleidaGr for data analysis.
Cell Assay
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.
Animal Protocol
Athymic Nu/Nu mice inoculated subcutaneously with VEGF-A-expressing C6 rat glioma cells
~4 mg/kg/day
Injection i.p.
References

[1]. J Biol Chem. 2007 May 4;282(18):13395-401.

[2]. Mol Biol Cell. 1996 Jan;7(1):25-42.

[3]. Cancer Res. 2005 Apr 15;65(8):3336-46.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C51H79NO13
Molecular Weight
914.18
Exact Mass
913.55514
Elemental Analysis
C, 67.01; H, 8.71; N, 1.53; O, 22.75
CAS #
53123-88-9
Appearance
White to off-white solid powder
SMILES
C[C@@H](C([C@H](OC)[C@H](O)/C(C)=C/[C@@H](C)C(C[C@@H]([C@H](C)C[C@@H]1CC[C@@H](O)C(OC)C1)OC2=O)=O)=O)C[C@H](C)/C=C/C=C/C=C(C)/[C@@H](OC)C[C@H]3O[C@](C(C(N4[C@H]2CCCC4)=O)=O)(O)[C@H](C)CC3
InChi Key
QFJCIRLUMZQUOT-PYYJPVDBSA-N
InChi Code
InChI=1S/C51H79NO13/c1-30-16-12-11-13-17-31(2)42(61-8)28-38-21-19-36(7)51(60,65-38)48(57)49(58)52-23-15-14-18-39(52)50(59)64-43(33(4)26-37-20-22-40(53)44(27-37)62-9)29-41(54)32(3)25-35(6)46(56)47(63-10)45(55)34(5)24-30/h11-13,16-17,25,30,32-34,36-40,42-44,46-47,53,56,60H,14-15,18-24,26-29H2,1-10H3/b13-11+,16-12+,31-17+,35-25+/t30-,32-,33-,34-,36-,37+,38+,39+,40-,42+,43+,44?,46-,47+,51-/m1/s1
Chemical Name
(3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34, 34a-hexadecahydro-9,27-dihydroxy-3-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-methoxycyclohexyl]-1-methylethyl]-10,21-dimethoxy-6,8,12,14,20,26-hexamethyl-23,27-epoxy-3H-pyrido[2,1-c][1,4] oxaazacyclohentriacontine-1,5,11,28,29 (4H,6H,31H)-pentone
Synonyms
AY 22989; AY22989; AY-22989; NSC-2260804; RAPA; RAP; RPM; SLM; AY 22989; SILA 9268A; WY090217; WY-090217; WY 090217; C07909; D00753; Rapamune
HS Tariff Code
2934.99.9001
Storage

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
DMSO: ~20 mg/mL (21.9 mM)
Water: <1 mg/mL
Ethanol: <1 mg/mL
Solubility (In Vivo)
2% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5 mg/mL
 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 1.0939 mL 5.4694 mL 10.9388 mL
5 mM 0.2188 mL 1.0939 mL 2.1878 mL
10 mM 0.1094 mL 0.5469 mL 1.0939 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

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Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
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Clinical Trial Information
NCT Number Status Interventions Conditions Sponsor/Collaborators Start Date Phases
NCT04996719 Active
Recruiting
Drug: Rapamune Heart Failure Mayo Clinic May 24, 2022 Phase 1
NCT05342519 Active
Recruiting
Drug: Rapamycin
Drug: Placebo
Vitiligo Medical University of
South Carolina
July 28, 2022 Phase 2
NCT04488601 Active
Recruiting
Drug: Rapamycin
Drug: Placebo
Aging AgelessRx January 1, 2020 Phase 2
NCT01529593 Active
Recruiting
Drug: Temsirolimus
Drug: Metformin
Advanced Cancers M.D. Anderson Cancer Center March 26, 2012 Phase 1
NCT01517243 Active
Recruiting
Drug: Sunitinib
Drug: Temsirolimus
Metastatic Renal Cell Carcinoma Dartmouth-Hitchcock Medical Center June 2010 Phase 2
Biological Data
  • Rapamycin (Sirolimus)

  • Rapamycin (Sirolimus)
  • Rapamycin (Sirolimus)
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