Temsirolimus

Biological Activity

Temsirolimus (CCI-779) inhibited the growth of xenografts derived from both cell lines with greater effects against PC-3 than DU145 tumors. Temsirolimus caused mild myelosuppression. The activity of mitoxantrone or docetaxel was limited, but CCI-779 given between courses of chemotherapy increased growth delay of PC-3 xenografts. High-dose temsirolimus inhibits mTOR signaling through an FKBP12-independent mechanism that leads to profound translational repression. This distinctive high-dose compound effect could be directly related to the antitumor activities of temsirolimus and other rapalogues in human cancer patients. The rapamycin analogue CCI-779 is an important new agent to investigate in the treatment of human brain tumors, particularly PNET/MB. The rapamycin analogue temsirolimus improved performance on four different behavioral tasks and decreased aggregate formation in a mouse model of Huntington disease.

Product Citations

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  Mol Cell. 2018 Dec 3.

  Phosphorylated RB Promotes Cancer Immunity by Inhibiting NF-κB Activation and PD-L1 Expression.
  Temsirolimus purchased from AbMole

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  Cell Death and Disease. 2018 Jan 26;9:137-52.

  Downregulation of MCL-1 and upregulation of PUMA using mTOR inhibitors enhance antitumor efficacy of BH3 mimetics in triple-negative breast cancer
  Temsirolimus purchased from AbMole

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  J Agric Food Chem. 2017 Mar 15;65(10):2089-2099.

  10-Gingerol, a Phytochemical Derivative from “Tongling White Ginger”, Inhibits Cervical Cancer: Insights into the Molecular Mechanism and Inhibitory Targets
  Temsirolimus purchased from AbMole

Customer Product Validations & Biological Datas

	Source	Cell Death & Disease (2018). Figure 2. Temsirolimus (Abmole Bioscience)
	Method	colony formation assays
	Cell Lines	MDA-MB-231, MDA-MB-157, and MDA-MB-468 cells
	Concentrations	1 μM
	Incubation Time	24 h
	Results	But treatment with Temsirolimus in combination with ABT263 or ABT199 only induced modest cell viability inhibition.

	Source	Cell Death & Disease (2018). Figure 1. Temsirolimus (Abmole Bioscience)
	Method	Immunoblotting analysis
	Cell Lines	TNBC cell lines
	Concentrations	1 μM
	Incubation Time	24 h
	Results	As expected, we found the mTOR inhibitors, especially the BEZ235 and AZD8055, could significantly inhibit the phosphorylation of AKT and 4EBP1 and efficiently led to decreasing MCL-1 expression.

	Source	APMIS.m (2017). Figure 7. temsirolimus (Abmole Bioscience Inc. Houston, TX, USA)
	Method	Western blot
	Cell Lines	HeLa cells
	Concentrations
	Incubation Time
	Results	In addition, exposure with deguelin and temsirolimus could weaken the inhibitory attributes of 10-G on the expression of p-AKT and p-mTOR.

Protocol

Cell Experiment
Cell lines	A549, H157, H460, H446, HCT116, HT29, SW480, DLD1, Caco2, LNCap, DU145, MDA468, MDA231, HEK293, and PC3-MM2
Preparation method	Exposing cells to various concentrations of Temsirolimus for 72 hours. After treatment, using CellTiter AQ assay kit to determing viable cell densities by MTS dye conversion.
Concentrations	Dissolved in DMSO, final concentrations ~20 μM
Incubation time	72 hours

Animal Experiment
Animal models	Female athymic nude mice injected s.c. with DAOY, or U251 cells
Formulation	Prepared in 100% EtOH as a 50 mg/mL stock solution, and diluted in 5% Tween 80 and 5% polyethylene glycol 400
Dosages	20 mg/kg
Administration	Injection daily 5 times per week

Conversion of different model animals based on BSA (Value based on data from FDA Draft Guidelines)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of resveratrol used for a mouse (22.4 mg/kg) to a dose based on the BSA for a rat, multiply 22.4 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for resveratrol of 11.2 mg/kg.

Chemical Information

References

A new pharmacologic action of CCI-779 involves FKBP12-independent inhibition of mTOR kinase activity and profound repression of global protein synthesis.
Shor B, et al. Cancer Res. 2008 Apr 15;68(8):2934-43. PMID: 18413763.

Effects of the mammalian target of rapamycin inhibitor CCI-779 used alone or with chemotherapy on human prostate cancer cells and xenografts.
Wu L, et al. Cancer Res. 2005 Apr 1;65(7):2825-31. PMID: 15805283.

Inhibition of mTOR induces autophagy and reduces toxicity of polyglutamine expansions in fly and mouse models of Huntington disease.
Ravikumar B, et al. Nat Genet. 2004 Jun;36(6):585-95. PMID: 15146184.

Antitumor activity of the rapamycin analog CCI-779 in human primitive neuroectodermal tumor/medulloblastoma models as single agent and in combination chemotherapy.
Geoerger B, et al. Cancer Res. 2001 Feb 15;61(4):1527-32. PMID: 11245461.