Rapamycin

Manufactured by Pfizer

Sourced in United States, Germany

Rapamycin is a macrolide compound with immunosuppressive and antiproliferative properties. It functions as an inhibitor of the mammalian target of rapamycin (mTOR) pathway, which plays a critical role in cellular growth and proliferation.

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Lab products found in correlation

Rapamycin, by Merck Group (2 mentions) Rapamune, by Pfizer (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) 4e bp1, by Cell Signaling Technology (2 mentions) Peroxidase conjugated secondary antibody, by Cell Signaling Technology (2 mentions) Microplate reader, by Thermo Fisher Scientific (2 mentions) 2500 gel imager, by Bio-Rad (1 mentions) Bca protein concentration assay kit, by Biosharp (1 mentions) 7900 fast, by Thermo Fisher Scientific (1 mentions) Sybr qpcr mix, by Thermo Fisher Scientific (1 mentions) Superscript 3 rt kit, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Y79 cells, by American Type Culture Collection (1 mentions) Recombinant human il 2, by Miltenyi Biotec (1 mentions) Clinimacs, by Miltenyi Biotec (1 mentions) X vivo 15 medium, by Lonza (1 mentions) Temsirolimus, by Pfizer (1 mentions) 5 fudr, by Merck Group (1 mentions) Cisplatin, by Merck Group (1 mentions) Erlotinib, by LC Laboratories (1 mentions)

17 protocols using rapamycin

Investigating Oxidative Stress in Y79 Cells

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Y79 cells [American Type Culture Collection (ATCC) (Manassas)], Rapamycin (Pfizer), enzyme-linked immunosorbent assay (ELISA) kits of reactive oxygen species (ROS) and malon- dialdehyde (MDA) (Nanjing Jiancheng Bioengineering Institute), radioimmunoprecipitation assay (RIPA) lysis buffer (Beyotime), loading buffer, protease inhibitor and bicinchoninic acid (BCA) protein concentration assay kit (Biosharp), glyceraldheyde 3-phosphate dehydrogenase (GAPDH) and secondary antibodies (ImmunoWay), primary antibodies (Cell Signaling Technology, Inc.), TRIzol reagent (Invitrogen; Thermo Fisher Scientific, Inc.), diethyl pyrocarbonate (DEPC)-treated water, SuperScript III RT kit and SYBR qPCR Mix (Applied Biosystems; Thermo Fisher Scientific, Inc.), electrophoresis apparatus (Bio-Rad Laboratories, Inc.), microplate reader (Thermo Fisher Scientific, Inc.), 2500 gel imager (Bio-Rad Laboratories, Inc.) and quantitative polymerase chain reaction (qPCR) instrument (7900 Fast, Applied Biosystems; Thermo Fisher Scientific, Inc.).

Yao J., Xu M, & Liu Z. (2020). Rapamycin inhibits proliferation and apoptosis of retinoblastoma cells through PI3K/AKT signaling pathway. Oncology Letters, 19(4), 2950-2956.

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Expansion of human regulatory T cells

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Peripheral blood from healthy donors was used for enrichment of natural regulatory T cells (nTreg). Two consecutive CliniMACS runs, compromising a CD8⁺ cell depletion followed by a CD25⁺ cell selection, were performed according to the manufacturer‘s instructions (Miltenyi Biotec). Target cell fraction was cultured in complete nTreg-medium, containing X-Vivo 15 medium (Lonza) supplemented with fetal calf serum (FCS, Hyclone), recombinant human IL-2 (Miltenyi Biotec) and Rapamycin (Pfizer). Depending on cell numbers, expansion process of Tregs was performed in 96-well plates and 24-well plates, respectively, for 23 days, using complete nTreg-medium and repetitive stimulation with anti-CD3/CD28 MACSiBead particles (Treg Activation/Expansion Kit, Miltenyi Biotec). Cells were collected for downstream DNA methylation analysis at several time-points during culture.

Ou K., Hamo D., Schulze A., Roemhild A., Kaiser D., Gasparoni G., Salhab A., Zarrinrad G., Amini L., Schlickeiser S., Streitz M., Walter J., Volk H.D., Schmueck-Henneresse M., Reinke P, & Polansky J.K. (2021). Strong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function. Frontiers in Cell and Developmental Biology, 9, 751590.

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Xenogeneic GVHD and Skin Rejection Model

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For xenogeneic graft‐versus‐host‐disease (GVHD) model, 1.5 × 10⁷ fresh human PBMC were intravenously injected in 1.5‐Gy‐irradiated NSG mice as previously described.¹⁰ Human PBMC engraftment was monitored in blood, and GVHD development was characterized by ≥20% body weight loss. For the skin rejection model, human skins were obtained from healthy donors from abdominoplasty surgery, and transplantation was performed as previously described.¹⁰ A total of 5.0 × 10⁶ PBMCs, allogeneic to the graft, were i.v. injected. A graft rejection score was established from 0 to 5 based on macroscopic observations: 1, the skin starts to peel off; 2, thick skin; 3, scab; 4, edges start to take off; 5, the skin is entirely gone. Osmotic pumps (Alzet, model 1004, Cupertino, CA) were filled with rhIL‐34 (.42 μg/h, i.e. .4 m/kg/d; Thermo Fisher, Waltham, MA or Preprotech, Neuilly‐Sur‐Seine, France) and placed i.p. on the day before the injection of the PBMC. Rapamycin (.4 mg/kg/d for 10 days, Rapamune, Pfizer) was injected intraperitoneally.

Freuchet A., Salama A., Bézie S., Tesson L., Rémy S., Humeau R., Règue H., Sérazin C., Flippe L., Peterson P., Vimond N., Usal C., Ménoret S., Heslan J., Duteille F., Blanchard F., Giral M., Colonna M., Anegon I, & Guillonneau C. (2022). IL‐34 deficiency impairs FOXP3+ Treg function in a model of autoimmune colitis and decreases immune tolerance homeostasis. Clinical and Translational Medicine, 12(8), e988.

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Rapamycin-Induced DiCre Recombinase Excision

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DiCre recombinase mediated excision of targeted DNA sequences in vivo was achieved by administration of 200 μg Rapamycin (1mg/ml stock, Rapamune, Pfizer) to mice by oral gavage, 24 hours prior to transmission to mosquitoes. In order to achieve excision in the mosquito stages, 10 μg Rapamycin (1 mg/ml stock solution in DMSO, Sigma-Aldrich) was added to 10 ml 10% sucrose solution and used to feed mosquitoes. The Rapamycin dose was refreshed every alternate day along with the sucrose solution.

Fernandes P., Briquet S., Patarot D., Loubens M., Hoareau-Coudert B, & Silvie O. (2020). The dimerisable Cre recombinase allows conditional genome editing in the mosquito stages of Plasmodium berghei. PLoS ONE, 15(10), e0236616.

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Dissolution and Formulation of COTI-2 and Other Anticancer Drugs

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COTI-2, supplied by Cotinga Pharmaceuticals (London, Ontario), was dissolved in 100% dimethyl sulfoxide (DMSO) stock solution and diluted in αMEM or DMEM plus 10% FBS so that final DMSO concentrations were 0.5–1.0%. For the AN3-CA xenograft studies, 235 mg of COTI-2 was mixed with 4.84 g of Captisol® (CydexPharmaceuticals, Lenexa, KS) in 10 ml of sterile water (Hospira, Inc., Lake Forest, IL). For the PANC-1 xenograft studies, COTI-2 was dissolved in phosphate-citrate buffer at pH 2.3.
Cisplatin, 5-FUdR (Sigma-Aldrich, St. Louis, MO), vinorelbine (GlaxoSmithKline, Brentford, UK), temsirolimus, rapamycin (Pfizer, New York, NY), gemcitabine (Eli Lilly, Indianapolis, IN), paclitaxel, carboplatin (Bristol-Myers Squibb Co., New York, NY), and vincristine were obtained from the London Health Sciences Centre Pharmacy (London Regional Cancer Program, London, Ontario). Cetuximab was purchased from ImClone Systems (New York, NY). Erlotinib was obtained from LC Laboratories (Woburn, MA).

Maleki Vareki S., Salim K.Y., Danter W.R, & Koropatnick J. (2018). Novel anti-cancer drug COTI-2 synergizes with therapeutic agents and does not induce resistance or exhibit cross-resistance in human cancer cell lines. PLoS ONE, 13(1), e0191766.

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Antibody Panel for Signaling Proteins

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Antibodies to detect MET (D1C2), mTOR (7C10), LC3B, PARP, Caspase 3, Cyclin D1, phospho-ALK (Tyr1604), phospho-MET (Tyr1234/1235), AKT, phospho-AKT (Ser473), phospho-MAPK (Tyr202/204), 4EBP1, phospho-4EBP1 (Thr37/46), S6K, phospho-S6K (Ser371), RPS6, phospho-RPS6 (Ser235/236), STAT3, phospho-STAT3 (Tyr705) were from Cell Signaling (Danvers, MA, USA). Anti-RIP3 was from Santa Cruz Biotechnology (Dallas, TX, USA), Anti-Ki-67 from Cell Marque (Rocklin, CA, USA), and Anti-β-Actin from Sigma-Aldrich (München, Germany). ALK-1A4 (immunoblotting) was from Origene (Rockville, MD, USA), ALK-D5F3 (immunohistochemistry) was from Ventana Medical Systems (Tucson, AZ, USA). Crizotinib and rapamycin were purchased from Pfizer (Berlin, Germany) and ChemieTek (Indianapolis, IN, USA), respectively.

Mönch D., Bode-Erdmann S., Kalla J., Sträter J., Schwänen C., Falkenstern-Ge R., Klumpp S., Friedel G., Ott G, & Kalla C. (2018). A subgroup of pleural mesothelioma expresses ALK protein and may be targetable by combined rapamycin and crizotinib therapy. Oncotarget, 9(29), 20781-20794.

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Rapamycin-Induced DiCre Recombination

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DiCre recombinase mediated excision of targeted DNA sequences in vivo was achieved by oral administration of 200 µg Rapamycin (1mg/ml stock, Rapamune, Pfizer) to mice, 24 hours prior to transmission to mosquitoes. In order to achieve excision in the mosquito stages, 10 µg Rapamycin (1 mg/ml stock solution in DMSO, Sigma-Aldrich) was added to 10 ml 10% sucrose solution and used to feed mosquitoes. The Rapamycin dose was refreshed every alternate day along with the sucrose solution.

Fernandes P., Briquet S., Patarot D., Loubens M., Hoareau-Coudert B., & Silvie O. (2020). The dimerisable Cre recombinase allows conditional genome editing in the mosquito stages of Plasmodium berghei.

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Ginkgo Biloba Extract Composition

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GBE powder was purchased from Beijing Handian Pharmaceutical Co. Ltd., atorvastatin and rapamycin were purchased from Pfizer Pharmaceuticals Co. Ltd., and streptozotocin (STZ) was purchased from Sigma Co. Ltd. Ginkgo flavonoid and terpenoid contents in GBE in the present are 44.9% and 6.3%, respectively, whereas the amount of ginkgo acid is limited to <1 ppm.

Tian J., Popal M.S., Liu Y., Gao R., Lyu S., Chen K, & Liu Y. (2019). Ginkgo Biloba Leaf Extract Attenuates Atherosclerosis in Streptozotocin-Induced Diabetic ApoE-/- Mice by Inhibiting Endoplasmic Reticulum Stress via Restoration of Autophagy through the mTOR Signaling Pathway. Oxidative Medicine and Cellular Longevity, 2019, 8134678.

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Rapamycin Dosage Optimization for T Cell Response

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Rapamycin (Pfizer, Apotex Corp., or Amneal Pharmaceuticals) was administered daily i.p. (600 μg per kg). In Supplemental Figure 3, 75 μg per kg of Rapamycin was administered i.p. Blood concentration with these doses has been described previously (31 (link)). 600 μg per kg of Rapamycin was used in most experiments, since this dose, although not statistically significant, showed a slightly better CD8⁺ T cell response compared with 75 μg per kg of Rapamycin. Control mice received vehicle.

Ando S., Perkins C.M., Sajiki Y., Chastain C., Valanparambil R.M., Wieland A., Hudson W.H., Hashimoto M., Ramalingam S.S., Freeman G.J., Ahmed R, & Araki K. (2023). mTOR regulates T cell exhaustion and PD-1–targeted immunotherapy response during chronic viral infection. The Journal of Clinical Investigation, 133(2), e160025.

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Islet Transplantation: Isolation, Purification, and Immunosuppression

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Islets were isolated from pancreata obtained from multi-organ donors, using a modified automated method, and were then purified by centrifugation on a discontinuous gradient as previously described (14 (link)). Islets were then transplanted intra-hepatically according to ABO matching. Islet-transplanted patients received the standard triple immunosuppressive regimen: anti-thymoglobulin (Thymoglobulin, Genzyme, Framingham, MA) as induction, followed by treatment with FK506 ([Astellas, Deerfield, IL]; target blood levels between 6 and 8 ng/ml) and/or Cyclosporine ([Novartis, Basel, Switzerland]; target blood level 100 ng/ml) and/or Rapamycin ([Pfizer, New York, NY]; 8-15 ng/ml) and/or Micophenolate ([Roche, Basel, Switzerland]; 2g/die) and prednisone ([Bruno Farmaceutici, Italy] 5–10 mg/day); Cyclosporine drug level was assessed by immunocolorimetric assay (Siemens, Munich, Germany), FK506 by liquid chromatography–mass spectrometry (). Steroids were tapered and then withdrawn within 6 months post-transplant. C-peptide level was assessed by immunofluorimetric assay (Tosoh, Tokyo, Japan); Hba1c level was assessed by high-performance liquid chromatography (Biorad, Hercules, CA); EIR (exogenous insulin requirement) was collected through patient interview.

Vergani A., Gatti F., Lee K.M., D’Addio F., Tezza S., Chin M., Bassi R., Tian Z., Wu E., Maffi P., Nasr M.B., Kim J.I., Secchi A., Markmann J.F., Rothstein D.M., Turka L.A., Sayegh M.H, & Fiorina P. (2014). TIM4 regulates the anti-islet Th2 alloimmune response. Cell transplantation, 24(8), 1599-1614.

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