Biological Activity

Bioactivity / Mechanism

20-Hydroxyecdysone induced EcRE-dependent transcription in Bombyx cells. Met1 knockdown reduced the response, including with EcR/USP co-expression, supporting a context-dependent role for MET in ecdysone signaling. [1]

Literature-reported observations and conditions; not Adooq-tested protocols or general treatment recommendations.

Target & Activity

Target	Activity	Value
Human Endogenous Metabolite

Cellular Effect

Cell Line	Activity	Value	Description	Reference
Huh-7	CC50	> 700 μM	Cytotoxicity against human HuH7 cells assessed as reduction in cell viability after 72 hrs by MTS assay	[PMID: 26988299]
Huh-7	EC50	53.55 μM	Antiviral activity against Dengue virus 2 DN454009A infected in human HuH7 cells assessed as inhibition of viral RNA level measured 72 hrs post infection by qRT-PCR method	[PMID: 26988299]

In Vitro Studies

In differentiated C2C12 myotubes, 20-hydroxyecdysone increased Akt Ser473 phosphorylation after 2 h. The response was significant at 0.1 µM; this treatment concentration is not a binding-affinity or potency estimate. [2]

20E-responsive EcRE transcription in Bombyx cells

Cell-based study · Bombyx mori DZNU-Bm-12 cells with EcRE luciferase reporter [1]

20E increased reporter activity; Met1 RNAi attenuated the response, including with EcR/USP co-expression. Basal activity was little affected.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
EcRE-driven luciferase activity	Relative luciferase activity Increased with 20E; reduced by Met1 RNAi (different letter groups: P<0.05, ANOVA).	[1]

Literature-Reported Treatment Conditions

Treatment concentration: 1 µM
Exposure duration: 6 h hormone exposure after 48 h RNAi/transfection
Method: Cell treatment and dual-luciferase assay
Reported limits: Solvent and lot purity not specified for this reporter experiment; replicate number and error-bar definition not explicit in its caption.

Methods and source locations

Compound reported in the paper: 20-Hydroxyecdysone; Compound label in the paper: 20E

Applies to the reported research material and model; no verification of current Adooq lots or clinical efficacy.

Figure 5E (Results prose calls it 5F); caption; Methods: RNAi and hormone treatment, Transient transfection assay, Luciferase assay

EcRE-driven luciferase activity: EcRE reporter with hsp70 mini-promoter; actin3-driven Renilla internal reference; egfp dsRNA and hormone-absent controls. — Figure 5E (Results prose calls it 5F); caption; Methods: RNAi and hormone treatment, Transient transfection assay, Luciferase assay

Concentration response of Akt phosphorylation in C2C12 myotubes

Cell-based study · Differentiated mouse C2C12 myotubes (ATCC CRL-1772), serum-starved overnight [2]

20HE increased Akt Ser473 phosphorylation after 2 h. The authors report a greater-than-threefold response at 0.1 µM and no significant difference between 1 µM and higher tested concentrations.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Akt Ser473 phosphorylation normalized to total Akt	Fold change versus vehicle >3-fold at 0.1 µM; P<0.05 versus control. No significant difference between 1 µM and higher tested concentrations.	[2]

Literature-Reported Treatment Conditions

Treatment concentration: 0.05, 0.1, 0.5, 1, 5 and 10 µM; vehicle control
Exposure duration: 2 h
Method: Treatment in serum-free medium
Vehicle: 0.1% ethanol
Reported limits: Reagent lot and purity not given.

Methods and source locations

Compound reported in the paper: 20-Hydroxyecdysone; Compound label in the paper: 20HE

Applies to the reported research material and model; no verification of current Adooq lots or clinical efficacy.

Figure 2 image/caption; Results: Akt Activation; Experimental: Materials, Cell Culture, Cell Signaling Experiments, Western Blot Analysis

Akt Ser473 phosphorylation normalized to total Akt: Western blot densitometry; mean ± SEM of three experiments. Methods specify four wells per treatment and pooling two wells per lysate; one-way ANOVA/Bonferroni. — Figure 2 image/caption; Results: Akt Activation; Experimental: Materials, Cell Culture, Cell Signaling Experiments, Western Blot Analysis

In Vivo Studies

Mouse muscle responses depended on the experimental setting. Continuous 20E infusion increased triceps mass at 5 days but not 15 days. In sedentary aging mice, 28-day oral treatment did not significantly alter measured muscle mass or anabolic signaling; a separate single-dose experiment found no significant signaling response at 90 min. These designs differ in age, route and timing; the cause of their different responses remains uncertain. [3,4]

Chronic oral 20E in sedentary aging mice

Animal study · 20-month-old male C57BL/6 mice; randomized CT, ATE or 20E groups (initial n=12 each) [4]

Daily oral 20E for 28 days did not significantly alter measured muscle mass, fiber morphology or anabolic signaling. Endpoints were sampled 24 h after the final treatment.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Muscle mass, fiber morphology and gastrocnemius anabolic signaling	Between-group significance No significant treatment differences; Akt Ser473 phosphorylation P=0.485; p70S6K Thr389 phosphorylation P=0.914.	[4]

Conditions shared by the study groups

Route: Oral, treated food pellet
Frequency: Once daily
Treatment course: 28 days
Vehicle: Ethanol evaporated from pellet

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
20E versus vehicle 50 mg/kg/day	Vehicle-treated pellet	Daily oral 20E for 28 days did not significantly alter measured muscle mass, fiber morphology or anabolic signaling. Endpoints were sampled 24 h after the final treatment.	[4]

20E versus vehicle: Model: 20-month-old male C57BL/6 mice; randomized CT, ATE or 20E groups (initial n=12 each)

Methods and source locations

Compound reported in the paper: 20-Hydroxyecdysone; Compound label in the paper: 20E

Applies to the reported research material and model; no verification of current Adooq lots or clinical efficacy.

Sections 2.2.1, 2.3–2.6, 3.1–3.4; Tables 1–4; Figure 2

Muscle mass, fiber morphology and gastrocnemius anabolic signaling: Muscle wet weights; immunofluorescence morphology; Western blot; mean ± SEM; signaling one-way ANOVA. Figure 2 values normalized to CT. — Sections 2.2.1, 2.3–2.6, 3.1–3.4; Tables 1–4; Figure 2

20E versus vehicle · Sections 2.2.1, 2.3–2.6, 3.1–3.4; Tables 1–4; Figure 2

Acute oral 20E and muscle signaling in aging mice

Animal study · 20-month-old male C57BL/6J mice; 20E n=7, vehicle n=6; separate ATE n=7 arm [4]

A single oral 20E dose did not significantly alter the measured gastrocnemius phosphorylation markers at 90 min.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Gastrocnemius phosphorylation of Akt, p70S6K, 4EBP-1 and rpS6	Between-group significance No significant differences: Akt P=0.943; p70S6K P=0.653; 4EBP-1 P=0.234; rpS6 P=0.953.	[4]

Conditions shared by the study groups

Route: Oral gavage
Frequency: Single dose
Treatment course: 90 min to sampling
Vehicle: Corn oil

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
20E versus vehicle 50 mg/kg	Corn oil vehicle	A single oral 20E dose did not significantly alter the measured gastrocnemius phosphorylation markers at 90 min.	[4]

20E versus vehicle: Model: 20-month-old male C57BL/6J mice; 20E n=7, vehicle n=6; separate ATE n=7 arm

Methods and source locations

Compound reported in the paper: 20-Hydroxyecdysone; Compound label in the paper: 20E

Applies to the reported research material and model; no verification of current Adooq lots or clinical efficacy.

Sections 2.2.2, 2.4, 2.6, 3.3; Figure 3

Gastrocnemius phosphorylation of Akt, p70S6K, 4EBP-1 and rpS6: Western blot, CT-normalized; mean ± SEM; one-way ANOVA. Reported P values compare all three groups, not a separate pairwise 20E-versus-CT test. — Sections 2.2.2, 2.4, 2.6, 3.3; Figure 3

20E versus vehicle · Sections 2.2.2, 2.4, 2.6, 3.3; Figure 3

Continuous 20E infusion and muscle mass in adult mice

Animal study · Adult male C57BL/6J mice, 12 weeks old at purchase; weight-balanced randomized groups, n=9–10 [3]

Triceps brachii mass increased after 5 days, but not after 15 days; other measured muscle masses did not differ. This limited response does not establish generalized muscle anabolism.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Triceps brachii wet mass	mg, mean ± standard error 5 days: 20E 115 ± 8 mg versus saline 88 ± 3 mg, P=0.01; 15 days: 20E 89 ± 5 mg versus saline 94 ± 8 mg, not significant.	[3]

Conditions shared by the study groups

Route: Subcutaneous osmotic-pump infusion
Frequency: Continuous
Treatment course: 5 or 15 days
Vehicle: 0.9% NaCl

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
20E versus saline 5 mg/kg/day	Saline-only pump	Triceps brachii mass increased after 5 days, but not after 15 days; other measured muscle masses did not differ. This limited response does not establish generalized muscle anabolism.	[3]

20E versus saline: Model: Adult male C57BL/6J mice, 12 weeks old at purchase; weight-balanced randomized groups, n=9–10

Methods and source locations

Compound reported in the paper: 20-Hydroxyecdysone; Compound label in the paper: 20E

Applies to the reported research material and model; no verification of current Adooq lots or clinical efficacy.

Materials and Methods: Animals; Results: Body Tissue Analysis; Table 1; Discussion

Triceps brachii wet mass: Dissection and tissue weighing; ANOVA comparisons at each time point; Table 1. — Materials and Methods: Animals; Results: Body Tissue Analysis; Table 1; Discussion

20E versus saline · Materials and Methods: Animals; Results: Body Tissue Analysis; Table 1; Discussion

Specifications

Manufacturer	Adooq Bioscience
CAS Number	5289-74-7
Formula	C₂₇H₄₄O₇
Molecular Weight	480.6
Synonyms	β-Ecdysone; Ecdysterone
SMILES	C[C@]12CC[C@H]3C(=CC(=O)[C@H]4[C@@]3(C[C@@H]([C@@H](C4)O)O)C)[C@@]1(CC[C@@H]2[C@](C)([C@@H](CCC(C)(C)O)O)O)O

Storage & Solubility

In Vitro Solubility

Reported Solubility

Solvent	Reported Solubility
DMSO	86 mg/mL (178.92 mM)
Water	Insoluble
Ethanol	86 mg/mL (178.92 mM)

Preparing Stock Solutions

Stock preparation is unavailable for this product molecular weight.

Calculated stock concentrations do not establish solvent solubility. Refer to the reported solubility data for experimental guidance.

In Vivo Preparation

Concentration: 28 mg/mL
Vehicle: 1% DMSO+30% polyethylene glycol+1% Tween 80

Storage

Store powder at -20°C, keep desiccated.
In powder form, the chemical is stable for 36 months.
In solution, store at -20°C and use within 1 month to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.

Research References

[1] Guo E, He Q, Liu S, et al. MET is required for the maximal action of 20-hydroxyecdysone during Bombyx metamorphosis. PLoS One (2012). PMID: 23300902 DOI: 10.1371/journal.pone.0053256 Source material
[2] Gorelick-Feldman J, Cohick W, Raskin I. Ecdysteroids elicit a rapid Ca2+ flux leading to Akt activation and increased protein synthesis in skeletal muscle cells. Steroids (2010). PMID: 20363237 DOI: 10.1016/j.steroids.2010.03.008 Source material
[3] Cheng DM, Kutzler LW, Boler DD, et al. Continuous infusion of 20-hydroxyecdysone increased mass of triceps brachii in C57BL/6 mice. Phytother Res (2013). PMID: 22495969 DOI: 10.1002/ptr.4679 Source material
[4] Lawrence MM, Zwetsloot KA, Arthur ST, et al. Phytoecdysteroids Do Not Have Anabolic Effects in Skeletal Muscle in Sedentary Aging Mice. Int J Environ Res Public Health (2021). PMID: 33418916 DOI: 10.3390/ijerph18020370 Source material

Publications

2 publications citing this product

Chunyan Fang, .et al. , Development, 2022, Jan 15;149(2):dev199841 PMID: 35088829
Miwako Toyofuku, .et al. , J Insect Physiol, 2021, Oct;134:104294 PMID: 34389412

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20-Hydroxyecdysone

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