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Cycloheximide Protein Synthesis inhibitor

Name: Cycloheximide
Brand: Selleck Chemicals
SKU: S7418
Availability: InStock
Rating: 5 (515 reviews)

Cat.No.S7418

Cycloheximide, an antifungal antibiotic, is an eukaryote protein synthesis inhibitor with IC50 of 532.5 nM and 2880 nM for protein synthesis and RNA synthesis in vivo, respectively. Cycloheximide suppresses ferroptosis and inhibits autophagy.Cycloheximide (NSC-185) can be used to induce animal models of Dysmnesia.Solutions are unstable and should be fresh-prepared.This product is a hazardous chemical (acute toxicity/flammable/skin corrosive). Please use it while wearing a protective face mask, gloves, and clothing.

Chemical Structure

Molecular Weight: 281.35

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Quality Control

Batch: Purity: 99.96%

99.96

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Cell Culture, Treatment & Working Concentration

Cell Lines	Assay Type	Concentration	Incubation Time	Activity Description	PMID
CEM	Anticancer assay			Tested in vitro for anticancer activity against CEM cells, IC50 = 0.12 μM.	10072683
9L	Anticancer assay			Tested in vitro for anticancer activity against 9L cells, IC50 = 0.2 μM.	10072683
SK-MEL-28	Anticancer assay			Tested in vitro for anticancer activity against SK-MEL-28 cells, IC50 = 1 μM.	10072683
Vero	Cytotoxicity assay			Tested in vitro for cytotoxicity against Vero cells, IC50 = 1.2 μM.	10072683
LNCaP	Anticancer assay			Tested in vitro for anticancer activity against LNCaP cells, IC50 = 1.2 μM.	10072683
MCF-7	Anticancer assay			Tested in vitro for anticancer activity against MCF-7 cells, IC50 = 1.5 μM.	10072683
PBM	Cytotoxicity assay			Tested in vitro for cytotoxicity against PBM cells, IC50 = 2.1 μM.	10072683
HepG2	Anticancer assay			Tested in vitro for anticancer activity against HepG2 cells, IC50 = 2.5 μM.	10072683
SK-MES-1	Anticancer assay			Tested in vitro for anticancer activity against SK-MES-1 cells, IC50 = 2.7 μM.	10072683
PC-3	Anticancer assay			Tested in vitro for anticancer activity against PC-3 cells, IC50 = 3.5 μM.	10072683
Jurkat T-cells	Function assay			Inhibitory concentration against Human Jurkat T cells, IC50 = 0.93 μM.	10212121
Jurkat T-cells	Function assay			Inhibitory concentration against Human Jurkat T cells, IC80 = 1.54 μM.	10212121
Jurkat T-cells	Function assay		48 hours	Inhibitory concentration was evaluated on human Jurkat T-cells after their exposure for 48 hours, IC50 = 0.93 μM.	11052798
Jurkat T-cells	Function assay		48 hours	Inhibitory concentration was evaluated on human Jurkat T-cells after their exposure for 48 hours, IC80 = 1.54 μM.	11052798
CEM	Cytotoxicity assay			Cytotoxicity was determined in CEM cells, relative to RVT, IC50 = 0.08 μM.	15081000
PBM	Cytotoxicity assay			Cytotoxicity was determined in PBM cells, relative to RVT, IC50 = 0.46 μM.	15081000
Vero	Cytotoxicity assay			Cytotoxicity was determined in Vero cells, relative to RVT, IC50 = 0.53 μM.	15081000
HeLa	Function assay			Inhibition of hypoxia-induced HIF1 activation in human HeLa cells by luciferase reporter gene assay, IC50 = 0.036 μM.	15974627
medulloblastoma cells	Antiproliferative assay			Antiproliferative activity against mouse medulloblastoma cells harboring heterozygous ptch1 gene by MTT assay, EC50 = 0.042 μM.	17417631
neural precursor cells	Antiproliferative assay			Antiproliferative activity against mouse neural precursor cells by colony formation assay, EC50 = 0.054 μM.	17417631
astrocyte cells	Antiproliferative assay			Antiproliferative activity against mouse astrocyte cells by MTT assay, EC50 = 0.071 μM.	17417631
neural precursor cells	Antiproliferative assay			Antiproliferative activity against mouse neural precursor cells by MTT assay, EC50 = 0.142 μM.	17417631
HeLa	Cytotoxicity assay		24 hrs	Cytotoxicity against human HeLa cells after 24 hrs, CD50 = 0.1 μM.	18394884
3T3	Cytotoxicity assay		72 hrs	Cytotoxicity against mouse 3T3 cells after 72 hrs by MTT assay, IC50 = 0.912 μM.	18771242
HeLa	Function assay		2 hrs	Inhibition of protein synthesis in human HeLa cells assessed as [35S]cysteine/methionine utilization after 2 hrs by scintillation spectroscopy, IC50 = 0.5325 μM.	20118940
HeLa	Function assay		2 hrs	Inhibition of transcriptional activity in human HeLa cells assessed as [3H]uridine utilization after 2 hrs by scintillation counting, IC50 = 2.8801 μM.	20118940
3T3	Cytotoxicity assay		72 hrs	Cytotoxicity against mouse 3T3 cells after 72 hrs by MTT assay, IC50 = 0.26 μM.	20356064
BESM	Function assay		88 hrs	Antitrypanosomal activity against Trypanosoma cruzi amastigotes infected in BESM cells measured after 88 hrs postinfection by HTS assay, EC50 = 0.4 μM.	20547819
NIH/3T3	Cytotoxicity assay		48 hrs	Cytotoxicity against mouse NIH/3T3 cells after 48 hrs by MTT assay, IC50 = 1.1 μM.	21899268
MDA-MB-231	Cytotoxicity assay		48 hrs	Cytotoxicity against human MDA-MB-231 cells after 48 hrs by MTT assay, IC50 = 1.2 μM.	21899268
AGS	Cytotoxicity assay		48 hrs	Cytotoxicity against human AGS cells after 48 hrs by MTT assay, IC50 = 3.6 μM.	21899268
HT-29	Cytotoxicity assay		48 hrs	Cytotoxicity against human HT-29 cells after 48 hrs by MTT assay, IC50 = 12.8 μM.	21899268
HepG2-A16-CD81	Function assay	10 uM		NOVARTIS: Antimalarial liver stage activity measured as reduction in Plasmodium yoelii schizont area in HepG2-A16-CD81 cells by immuno-fluorescence, and median schizont size at 10uM compound concentration, IC50 = 0.0781 μM.	22096101
3T3	Cytotoxicity assay		72 hrs	Cytotoxicity against mouse 3T3 cells after 72 hrs by MTT assay, IC50 = 0.26 μM.	22437110
HepG2	Function assay			HARVARD: Inhibition of liver stage Plasmodium berghei infection in HepG2 cells, IC50 = 0.047 μM.	22586124
PA1	Cytotoxicity assay		24 hrs	Cytotoxicity against human PA1 cells after 24 hrs by MTT assay, IC50 = 40.6 μM.	23202484
PA1	Growth inhibition assay		24 hrs	Growth inhibition of human PA1 cells after 24 hrs by MTT assay, IC50 = 40.6 μM.	28011220
3T3	Cytotoxicity assay			Cytotoxicity against mouse 3T3 cells by SRB assay, IC50 = 0.3 μM.	29247859
CEM	Cytotoxicity assay		5 days	Cytotoxicity against human CEM cells assessed as decrease in cell viability after 5 days by MTT assay, IC50 = 0.2 μM.	29778528
Vero	Cytotoxicity assay		3 days	Cytotoxicity against African green monkey Vero cells assessed as decrease in cell viability after 3 days by MTT assay, IC50 = 0.2 μM.	29778528
3T3	Cytotoxicity assay		48 hrs	Cytotoxicity against rat 3T3 cells after 48 hrs by MTT assay, IC50 = 0.61 μM.	30146096
MCF7	Function assay	10 ug/mL	1 hr	Inhibition of HIF1alpha RNA translation in human MCF7 cells at 10 ug/mL pretreated for 1 hr prior to metabolic labeling by SDS-PAGE analysis	22607231
AGS	Apoptosis assay	150 ug/mL	48 hrs	Induction of apoptosis in human AGS cells assessed as early apoptosis level at 150 ug/mL after 48 hrs by Annexin V-FITC apoptosis assay	21899268
TC32	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for TC32 cells	29435139
A673	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for A673 cells	29435139
DAOY	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for DAOY cells	29435139
Saos-2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Saos-2 cells	29435139
BT-37	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-37 cells	29435139
RD	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for RD cells	29435139
SK-N-SH	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-SH cells	29435139
BT-12	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for BT-12 cells	29435139
NB1643	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB1643 cells	29435139
OHS-50	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for OHS-50 cells	29435139
BT-12	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-12 cells	29435139
LAN-5	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for LAN-5 cells	29435139
Rh41	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for Rh41 cells	29435139
SK-N-SH	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-SH cells	29435139
A673	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for A673 cells)	29435139
BT-37	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for BT-37 cells	29435139
Rh30	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for Rh30 cells	29435139
U-2 OS	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for U-2 OS cells	29435139
OHS-50	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for OHS-50 cells	29435139
RD	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for RD cells	29435139
SJ-GBM2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SJ-GBM2 cells	29435139
SK-N-MC	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for SK-N-MC cells	29435139
NB-EBc1	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Primary screen for NB-EBc1 cells	29435139
SK-N-MC	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SK-N-MC cells	29435139
SJ-GBM2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for SJ-GBM2 cells	29435139
TC32	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Confirmatory screen for TC32 cells	29435139
SJ-GBM2	qHTS assay			qHTS of pediatric cancer cell lines to identify multiple opportunities for drug repurposing: Orthogonal 3D viability screen for SJ-GBM2 cells	29435139
T47D	Function assay	10 uM	4 hrs	Inhibition of 1,10-phenanthroline-induced HIF1alpha activation in human T47D cells at 10 uM after 4 hrs by Western blotting	15974627
T47D	Function assay	0.3 uM	4 hrs	Inhibition of 1,10-phenanthroline-induced HIF1alpha activation in human T47D cells at 0.3 uM after 4 hrs by Western blotting	15974627
T47D	Function assay	3 uM	16 hrs	Inhibition of hypoxia-induced HIF1 activation in human T47D cells at 3 uM after 16 hrs by luciferase reporter gene assay	15974627
T47D	Function assay	0.7 uM	16 hrs	Inhibition of 1,10-phenanthroline-induced HIF1 activation in human T47D cells at 0.7 uM after 16 hrs by luciferase reporter gene assay	15974627
T47D	Function assay	10 uM	16 hrs	Inhibition of hypoxia-induced VEGF expression in human T47D cells at 10 uM after 16 hrs by ELISA	15974627
T47D	Function assay	0.3 uM	4 hrs	Inhibition of hypoxia-induced HIF1alpha activation in human T47D cells at 0.3 uM after 4 hrs by Western blotting	15974627
T47D	Function assay	10 uM	4 hrs	Inhibition of hypoxia-induced HIF1alpha activation in human T47D cells at 10 uM after 4 hrs by Western blotting	15974627
neural precursor cells	Function assay	3 uM		Induction of neurosphere phenotype changes in mouse neural precursor cells at 3 uM	17417631
HeLa	Cytotoxicity assay			Cytotoxicity against human HeLa cells assessed as inhibition of DNA replication by imaging analysis	18066055
HeLa	Function assay			Inhibition of mitosis in human HeLa cells by imaging analysis	18066055
HEK293T	Function assay	50 uM	30 mins	Effect on polyribosome profiling in human HEK293T cells assessed as depletion of polysomes at 50 uM after 30 mins by spectrophotometry	20118940
T47D	Antiproliferative assay	10 uM	48 hrs	Antiproliferative activity against human T47D cells at 10 uM after 48 hrs by SRB assay	23434131
MDA-MB-231	Antiproliferative assay	10 uM	48 hrs	Antiproliferative activity against human MDA-MB-231 cells at 10 uM after 48 hrs by SRB assay	23434131
NCI-H460	Function assay	10 ug/mL	up to 9 hrs	Reduction of HSP1 stability in human NCI-H460 cells assessed as protein degradation at 10 ug/mL up to 9 hrs by Western blot analysis	24746225
HeLa	Cytotoxicity assay	25 uM	18 hrs	Cytotoxicity against human HeLa cells assessed as reduction in cell viability at 25 uM after 18 hrs by inverse MTT assay	25028062
RAW264.7	Function assay	70 uM	6 hrs	Inhibition of luminescence emission in mouse RAW264.7 cells transfected with luciferase plasmid containing universal promoter PKG at 70 uM after 6 hrs by luciferase reporter gene assay	25667960
Click to View More Cell Line Experimental Data

Solubility

In vitro
Batch:

DMSO : 56 mg/mL (199.04 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 56 mg/mL

Water : 15 mg/mL

Molarity Calculator

Mass	Concentration	Volume	Molecular Weight
Mass value Mass unit	Concentration value Concentration unit	Volume value Volume unit	Molecular weight (g/mol)

Dilution Calculator Molecular Weight Calculator

In vivo batch selection In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

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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Average weight of animals: g

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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.

Chemical Information, Storage & Stability

Molecular Weight	281.35	Formula	C₁₅H₂₃NO₄	Storage (From the date of receipt)	3 years-20°C powder
CAS No.	66-81-9	--		Storage of Stock Solutions	1 year-80°Cin solvent 1 month-20°Cin solvent
Synonyms	NSC-185, Actidione, Naramycin A, CHX, FT 3422-2, NM-MCD 80			Smiles	CC1CC(C(=O)C(C1)C(CC2CC(=O)NC(=O)C2)O)C

Mechanism of Action

In vitro	Cycloheximide, a protein synthesis inhibitor, reduces GSIV-induced host cell death by attenuating phosphatidylserine exposure and loss of ΔΨm from apoptosis/necrosis.
In vivo	This compound, a protein synthesis inhibitors, acts on memory by altering modulators of memory formation as a secondary consequence of the inhibition of protein synthesis.
References	[1] https://pubmed.ncbi.nlm.nih.gov/26548846/ [2] https://pubmed.ncbi.nlm.nih.gov/22610049/ [3] https://pubmed.ncbi.nlm.nih.gov/5541758/ [4] https://pubmed.ncbi.nlm.nih.gov/30916348/

Applications

Methods	Biomarkers	PMID
Western blot	β-catenin / MSX2 LIFR / gp130 / TfR BIP / P-P70S6K / P62 / ATF4 / Actin / LC3-I / LC3-II CYP87A3 ERK3 / IRP2	12095419
RNA blot	PDI-2 / tubulin / rRNA LEF1 / Cyc-D1 / IGF-1 / HGF / VEGF / KGF / GAPDH	18713834
Immunofluorescence	SCD6 / DHH1 / XRNA α-eIF3 / α-Dcp1 TUNEL Rpl25 / Nhp6 β-Catenin	18713834

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number	Recruitment	Conditions	Sponsor/Collaborators	Start Date	Phases
NCT03700788	Not yet recruiting	Apical Periodontitis	University of Southern California	May 30 2023	Phase 3
NCT05506566	Recruiting	Tumor\|Positron-Emission Tomography	First Affiliated Hospital of Fujian Medical University	May 1 2022	Phase 1\|Phase 2
NCT02202304	Withdrawn	Periodontal Disease\|Caries	Rosa Moreno Lopez\|Ivoclar Vivadent AG\|University of Aberdeen	September 10 2017	Phase 4
NCT01521325	Completed	Mesothelioma\|Pancreatic Cancer\|Ovarian Cancer\|Non-small Cell Lung Cancer	Morphotek	September 2011	Phase 1
NCT02168374	Completed	Dental Caries	Aline R F de Castilho\|Fundação de Amparo à Pesquisa do Estado de São Paulo\|University of Campinas Brazil	March 2008	Phase 2

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