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20-Hydroxyecdysone

20-Hydroxyecdysone, also known as ecdysterone, is a naturally occurring ecdysteroid hormone characterized by its polyhydroxylated ketosteroid structure and the molecular formula C₂₇H₄₄O₇.^[1] It serves as the principal molting hormone in arthropods, regulating critical developmental processes such as ecdysis, metamorphosis, and reproduction by binding to nuclear receptors and modulating gene expression.^[2] In plants, where it occurs as a phytoecdysteroid, it functions primarily as a chemical defense mechanism against herbivorous insects and pathogens, deterring feeding and inducing metabolic disruptions in non-adapted consumers.^[3] Chemically, 20-hydroxyecdysone features a cyclopentanoperhydrophenanthrene skeleton with a cis A/B ring junction, a 7-en-6-one chromophore, and multiple hydroxyl groups, including one at the 20-position that distinguishes it from its precursor, ecdysone.^[3] First isolated from the silk moth Bombyx mori in the 1960s, it is biosynthesized from cholesterol via the mevalonate pathway in both arthropods and plants, with over 400 related ecdysteroids identified to date.^[2] In arthropods, its levels fluctuate in precise pulses—typically in the nanomolar to micromolar range—to orchestrate stage-specific transitions, such as from larva to pupa, through pathways involving early and late gene responses.^[2] Beyond its ecological roles, 20-hydroxyecdysone has garnered attention for potential therapeutic applications in mammals, where it exhibits anabolic effects by enhancing protein synthesis and muscle growth without androgenic or estrogenic activity, activating pathways like PI3K/AKT/mTOR.^[1] Studies suggest benefits in treating neuromuscular disorders like sarcopenia and Duchenne muscular dystrophy, as well as cardio-metabolic conditions including prediabetes and obesity, with ongoing clinical trials evaluating its efficacy.^[2] Additionally, it demonstrates antioxidant, anti-inflammatory, and antineoplastic properties, positioning it as a candidate for respiratory diseases and stress-related ailments, though human safety and optimal dosing require further validation.^[3] Commercially, it is extracted from high-yielding plant sources such as Cyanotis arachnoidea and Rhaponticum carthamoides, which can contain up to 5% dry weight.^[2]

Chemical properties

Structure and nomenclature

20-Hydroxyecdysone is a polyhydroxylated ecdysteroid with the molecular formula C_{27}H_{44}O_7 and a molecular weight of 480.65 g/mol.^[1] Its structure consists of a tetracyclic steroid backbone derived from cholesterol, featuring rings A, B, C, and D fused in a cholestane configuration, with a ketone functional group at carbon 6 and a double bond between carbons 7 and 8 in ring B.^[4] Hydroxyl groups are attached at positions 2 (β), 3 (β), 14 (α), 20 (R), 22 (R), and 25, contributing to its polarity and solubility characteristics.^[5] The molecule exhibits specific stereochemistry at its chiral centers, defined as (2β,3β,5β,14α,20R,22R).^[4] The systematic IUPAC name for 20-hydroxyecdysone is (2β,3β,5β,14α,20R,22R)-2,3,14,20,22,25-hexahydroxycholest-7-en-6-one.^[5] It is commonly referred to by synonyms such as ecdysterone, β-ecdysone, or simply 20E in scientific literature.^[1] In comparison to the related ecdysteroid ecdysone, 20-hydroxyecdysone includes an additional hydroxyl group at position 20 on the side chain, resulting in one more oxygen atom and distinguishing its structural profile.^[6]

Physical and chemical characteristics

20-Hydroxyecdysone appears as a white to off-white crystalline powder.^[7]^[8] It exhibits moderate solubility in water (approximately 10 mg/mL), but is readily soluble in organic solvents such as ethanol (around 25 mg/mL), DMSO (around 30 mg/mL), and DMF (around 30 mg/mL).^[9] The compound's multiple hydroxyl groups contribute to its pKa values, with the strongest acidic pKa approximately 13.3.^[4] 20-Hydroxyecdysone is stable when stored dry in the dark at room temperature for extended periods (at least 4 years), but it is sensitive to light, heat, and strong acids or bases.^[10]^[9] Optimal storage conditions involve -20°C in an inert atmosphere to maintain integrity.^[7] Key spectroscopic properties include a UV absorption maximum at 243 nm, attributable to the 14α-hydroxy-7-en-6-one chromophore in the B-ring.^[9] In mass spectrometry, it shows a characteristic protonated molecular ion at m/z 481 [M+H]⁺.^[1] NMR spectra confirm the structure, with detailed ¹H and ¹³C assignments available from reference standards.^[1] Isolation from natural sources typically involves basic extraction techniques, such as refluxing plant material in ethanol, followed by purification using macroporous resins and recrystallization to achieve high purity (>97%).^[10]

Biosynthesis and natural occurrence

Biosynthesis in arthropods

20-Hydroxyecdysone (20E) biosynthesis in arthropods begins with dietary cholesterol as the obligate precursor, which is absorbed and transported to specialized endocrine glands. Arthropods lack the ability to synthesize sterols de novo, relying instead on exogenous sources for cholesterol. The initial conversion of cholesterol to 7-dehydrocholesterol is catalyzed by the Rieske-domain oxygenase Neverland (Nvd), a rate-limiting step that introduces a double bond at the C7-C8 position.^[11]^[12]^[13] Subsequent modifications involve a series of hydroxylations mediated by cytochrome P450 enzymes encoded by the Halloween genes, transforming 7-dehydrocholesterol through intermediates such as 5β-cholane-3α,7α-diol and others into ecdysone, the immediate precursor to 20E. These enzymes include Spook (CYP307A1) for early oxidation steps, Phantom (CYP306A1) for 25-hydroxylation, Disembodied (CYP302A1) for 22-hydroxylation, and Shadow (CYP315A1) for 2β-hydroxylation, culminating in the final 20α-hydroxylation of ecdysone to 20E by Shade (CYP314A1).^[14]^[15]^[16] This multi-step pathway ensures the production of the active molting hormone, with variations in enzyme orthologs observed across arthropod lineages, such as in crustaceans where some CYP genes may be compensated by paralogs.^[17] In insects, 20E is primarily synthesized in the prothoracic glands, while in crustaceans, the Y-organs serve as the main site of ecdysteroid production; synthesis occurs in pulses aligned with molting cycles, peaking during premolt stages to trigger exoskeleton renewal.^[18]^[19] Biosynthesis is tightly regulated by neuropeptides: in insects, prothoracicotropic hormone (PTTH) from the brain stimulates the prothoracic glands via cyclic AMP signaling to initiate ecdysone release, while negative feedback through the ecdysone receptor (EcR) in PTTH neurons modulates timing.^[20]^[21] In crustaceans, molt-inhibiting hormone (MIH) from the eyestalk sinus gland suppresses Y-organ activity, with de-repression driving synthesis peaks.^[22]^[23] Genetic studies provide strong evidence for these pathways, as mutations or RNAi knockdown of Neverland or Halloween genes result in developmental arrest due to ecdysteroid deficiency, phenotypes rescuable by exogenous 20E or precursors like 7-dehydrocholesterol.^[11]^[24]^[14] For instance, shade mutants fail to produce 20E, halting metamorphosis, underscoring the essential role of these enzymes in arthropod post-embryonic development.^[16]

Sources in plants and other organisms

20-Hydroxyecdysone, a prominent phytoecdysteroid, occurs naturally in numerous plant species, particularly within families such as Caryophyllaceae and Amaranthaceae. In Caryophyllaceae, genera like Silene exhibit high accumulation, with levels reaching 1–2% of dry weight in some species. Amaranthaceae members, including Achyranthes species such as A. faurieri and A. bidentata, also contain significant amounts, often up to 1.74% dry weight in roots. Spinacia oleracea (spinach), another Amaranthaceae representative, shows variable concentrations, typically ranging from 0.52 to 428 µg/g dry weight, though totals for ecdysteroids in plants can approach 3% under optimal conditions. Additionally, Cyanotis species like C. arachnoidea are notable sources, with roots accumulating up to 5.5% dry weight. These phytoecdysteroids serve as defensive compounds in plants, deterring herbivorous insects by disrupting their molting and feeding behaviors, thereby reducing phytophagous damage.^[25]^[2]^[26]^[27]^[28]^[29] Concentrations of 20-hydroxyecdysone vary markedly across plant organs and are influenced by environmental factors. Highest levels are often found in roots and seeds, as seen in Achyranthes japonica, where root and floral parts exceed stem and leaf concentrations during reproductive stages, and in Chenopodium album, with peaks in anthers, young leaves, and seeds. Stress conditions, such as salinity, induce accumulation; for instance, in Spinacia oleracea shoot cultures, 20-hydroxyecdysone reaches 0.20 mg/g dry weight at 200 mM NaCl, aiding membrane protection. Other stressors like jasmonates, physical damage, temperature fluctuations, and seasonal changes further modulate levels, with higher yields in spring shoots and autumn roots of perennials. Over 500 phytoecdysteroid analogues have been identified across more than 100 plant families, reflecting diverse accumulation patterns.^[30]^[26]^[31]^[25] Beyond plants, 20-hydroxyecdysone appears in trace amounts in other organisms, often linked to dietary exposure or environmental transfer. Limited reports indicate presence in algae, fungi, and ferns, though at low levels compared to angiosperms. In bacteria, biotransformations occur but natural endogenous production is not well-documented. Vertebrates acquire traces primarily through plant-based diets, as evidenced by metabolite detection in urine following ingestion of ecdysteroid-rich foods. Commercially, 20-hydroxyecdysone is extracted from high-yield sources like Ajuga turkestanica (aerial parts) and Leuzea carthamoides (syn. Rhaponticum carthamoides, roots at 1–2% dry weight), used in supplements and pharmacological preparations. Evolutionarily, phytoecdysteroids are proposed as ancient defense mechanisms, originating in ferns around the Devonian period (approximately 400 million years ago), predating arthropod diversification and functioning as antifeedants or endocrine disruptors against early herbivores.^[26]^[32]^[2]^[33]^[34] Analytical detection of 20-hydroxyecdysone in plant extracts relies on high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS), enabling sensitive quantification of low levels (limits of detection 6–80 ng/g). This method involves solid-liquid extraction with methanol-ethanol-water mixtures followed by solid-phase extraction cleanup, achieving recoveries of 75–99% and precision below 9% relative standard deviation. It simultaneously identifies multiple ecdysteroids, such as 20-hydroxyecdysone alongside makisterone A, supporting ecological and pharmaceutical research.^[27]

Biological functions

Role in arthropod development

20-Hydroxyecdysone (20E) functions as the principal steroid hormone regulating molting and metamorphosis in arthropods, triggering ecdysis by activating a nuclear receptor complex composed of the ecdysone receptor (EcR) and retinoid X receptor (RXR, known as ultraspiracle or USP in insects).^[35] This heterodimer binds to DNA response elements, initiating a transcriptional cascade that includes early genes such as Broad-Complex (BR-C), E74, E75, and E93, which coordinate downstream expression of genes for tissue remodeling, cuticle synthesis, and physiological adaptations.^[35] The pathway induces production of enzymes like proteases and chitinases essential for degrading the old exoskeleton during apolysis and facilitating new cuticle formation.^[36] In insects, 20E precisely times developmental transitions, such as the larval-to-pupal metamorphosis, where rising titers promote the histolysis of larval organs and proliferation of imaginal discs into adult structures.^[35] Titer fluctuations are critical: low levels during inter-molt periods support growth and maintenance, while periodic high pulses—often 7-fold elevations—signal commitment to molting, culminating in ecdysis behaviors mediated by neuropeptides like ecdysis-triggering hormone (ETH).^[37] In crustaceans, 20E coordinates vitellogenesis by elevating hemolymph levels that parallel ovarian maturation and yolk protein synthesis, with ovaries sequestering these hormones for oocyte provisioning.^[38] Across chelicerates, including ticks and scorpions, 20E supports analogous molting cycles and reproduction, though species like mites may prioritize ponasterone A as the dominant ecdysteroid.^[36] Experimental manipulations confirm 20E's mechanistic role; injections into insect larvae, such as Manduca sexta, induce dose-dependent molting and pupation by mimicking natural titer peaks.^[35] Conversely, RNAi-mediated knockdown of EcR in Drosophila melanogaster or Blattella germanica arrests development at molting stages, resulting in incomplete ecdysis and lethality due to failed cuticle separation.^[36] These findings underscore 20E's conserved function in synchronizing arthropod post-embryonic development across diverse taxa.^[36]

Functions in plants and other non-arthropods

In plants, 20-hydroxyecdysone functions primarily as a phytoecdysteroid, serving as a chemical defense mechanism against herbivorous insects by acting as a feeding deterrent.^[3] When ingested by non-adapted insects, it mimics the arthropod molting hormone, leading to premature or disrupted molting, reduced feeding, and impaired development and reproduction.^[39] For instance, exogenous application of 20-hydroxyecdysone to host plants significantly decreases larval feeding and survival in species like the diamondback moth (Plutella xylostella).^[40] This deterrent effect is concentration-dependent, with higher levels causing stronger avoidance behaviors in termites and other herbivores.^[41] Additionally, phytoecdysteroids exhibit allelochemical activity that enhances plant tolerance to pests such as nematodes, potentially inhibiting their growth and reproduction through similar hormonal interference.^[42] Unlike in arthropods, 20-hydroxyecdysone lacks endogenous hormonal activity or receptor-mediated signaling in plants, indicating its role is strictly defensive rather than regulatory for plant physiology.^[26] Experimental bioassays have demonstrated reduced insect feeding and herbivory on plants enriched with high levels of 20-hydroxyecdysone, such as spinach (Spinacia oleracea), where inducible production following damage further amplifies this protection.^[43] Phytoecdysteroids like 20-hydroxyecdysone are thought to have evolved as part of a plant-animal chemical warfare adaptation, allowing plants to exploit the hormonal vulnerabilities of invertebrate herbivores for ecological advantage.^[44] In non-arthropod invertebrates, such as nematodes and annelids, 20-hydroxyecdysone plays limited physiological roles, often related to exogenous exposure rather than endogenous production. In plant-parasitic nematodes, dietary uptake from host plants disrupts molting and development, serving as a novel defense mechanism without evidence of an intrinsic regulatory function in the nematodes themselves.^[45] For annelids, 20-hydroxyecdysone acts as an active molting hormone that supports growth and regenerative processes, with confirmed C-20 hydroxylation similar to arthropod pathways.^[46] Trace amounts of 20-hydroxyecdysone can transfer through dietary sources to vertebrates, but no clear endogenous function has been established in these organisms.^[47] Beyond defense, 20-hydroxyecdysone may contribute to stress responses in certain plants, particularly as an antioxidant in heavy metal-exposed seedlings, where it enhances ascorbate-glutathione cycle activity and protects against oxidative damage.^[48] In extremophile or stress-adapted plants, it potentially serves as an osmoprotectant, aiding tolerance to abiotic stresses like salinity or drought by stabilizing cellular functions, though this role remains secondary to its anti-herbivore effects.^[26]

Effects in mammals

Mechanisms of action

In mammals, 20-hydroxyecdysone (20E) does not bind to the ecdysone receptor (EcR), which is absent in mammalian cells, distinguishing its actions from those in arthropods where EcR drives developmental signaling. Instead, 20E exerts effects through interactions with mammalian receptors, including activation of estrogen receptor beta (ERβ) and the Mas1 receptor, a G protein-coupled receptor (GPCR) belonging to the Mas-related G protein-coupled receptor (Mrgpr) family.^[49] ERβ involvement is evidenced by 20E-induced transactivation of reporter genes in ERβ-transfected HEK293 cells and prevention of myotube hypertrophy by ERβ-selective antagonists in C2C12 cells, although direct binding to ERβ has not been conclusively demonstrated.^[50] Mas1 activation occurs via 20E stimulation of angiotensin-(1-7) production, promoting anti-inflammatory and anabolic signaling.^[49] Additionally, 20E engages the PI3K/Akt/mTOR pathway, enhancing protein synthesis in skeletal muscle cells as shown by increased phosphorylation in C2C12 myotubes at concentrations around 1 μM.^[51] Genomic effects of 20E primarily occur through ERβ-mediated transcription, leading to upregulation of insulin-like growth factor 1 (IGF-1) expression and inhibition of myostatin, a negative regulator of muscle growth; for instance, 20E treatment elevates serum IGF-1 levels in male rats and reduces myostatin mRNA in C2C12 cells in a dose-dependent manner (0.001–10 μM).^[50]^[49] Mas1 activation further contributes to myostatin suppression via the renin-angiotensin-aldosterone system (RAAS) protective arm.^[49] These transcriptional changes support anabolic processes in muscle tissue. Non-genomic effects involve rapid signaling cascades, including elevation of intracellular calcium levels in C2C12 cells, which precedes Akt phosphorylation and increased protein synthesis within minutes of 20E exposure.^[52] 20E also activates AMP-activated protein kinase (AMPK) in mammalian liver and adipose tissue, promoting metabolic shifts such as reduced lipogenesis through increased AMPK phosphorylation. These membrane-initiated responses occur independently of nuclear receptors and highlight 20E's role in quick cellular adaptations. The response to 20E exhibits dose dependency, with low micromolar concentrations (e.g., 0.01–1 μM) promoting anabolic effects like protein synthesis, while higher doses may shift toward metabolic regulation; in vitro studies show biphasic glucose secretion responses in hepatocytes, peaking at intermediate doses. Recent investigations confirm 20E's activation of the RAAS protective arm via Mas1, yielding anti-fibrotic and anti-inflammatory outcomes in endothelial and muscle cells, as demonstrated in models of metabolic stress.^[49]

Physiological effects

20-Hydroxyecdysone exhibits metabolic effects in mammals, including enhanced protein synthesis and energy production. In skeletal muscle cells, it stimulates protein synthesis through activation of the PI3K/Akt pathway, leading to increased myotube hypertrophy without androgenic activity.^[53] It also promotes energy production by phosphorylating AMPK and acetyl-CoA carboxylase, which supports mitochondrial function and reduces fatty acid synthesis. Furthermore, 20-hydroxyecdysone boosts lipid metabolism by lowering plasma triglycerides and cholesterol levels while improving insulin sensitivity; in diet-induced obese mice, oral administration (10 mg/kg for 13 weeks) reduced body weight gain by 18%, adipose mass by 41%, and plasma insulin by 4.5-fold, alongside decreased expression of gluconeogenic enzymes like PEPCK.^[54] In terms of tissue-specific responses, 20-hydroxyecdysone promotes skeletal muscle hypertrophy by increasing fiber cross-sectional area in a muscle-type-specific manner; in rats, subcutaneous administration (5 mg/kg for 5 days) enlarged slow-twitch soleus fibers by up to 20% and fast-twitch fibers in the extensor digitorum longus by 15%, accompanied by elevated myonuclear numbers indicative of satellite cell activation.^[55] It provides neuroprotection through anti-apoptotic and antioxidant mechanisms, as demonstrated in models of neurotoxicity where it preserved neuronal viability against oxidative stress.^[56] Additionally, it exerts anti-inflammatory effects, including a 38% reduction in C-reactive protein in overweight patients with metabolic syndrome treated with 100 mg/day for 3 months.^[57] In high-fat-high-fructose-fed ovariectomized rats, 20E (10-20 mg/kg) improved metabolic and cardiovascular function via activation of AMPK (up to +262%) and increased FGF21 expression (up to +468%).^[58] The toxicity profile of 20-hydroxyecdysone is favorable, with low acute toxicity reported; the oral LD50 exceeds 9 g/kg in mice, and intraperitoneal LD50 is 6.4 g/kg in rodents, while no subacute toxicity occurs in rats at chronic doses up to 2 g/kg/day.^[59]^[57] At high doses, it may exhibit potential estrogenic effects, as muscle hypertrophy induced by 20-hydroxyecdysone is reversible by estrogen receptor antagonists, suggesting mediation via ERβ. Animal studies highlight functional improvements, including increased grip strength in rodents; in young rats treated orally with 20-hydroxyecdysone (5 mg/kg for 28 days), forelimb grip strength rose by 18% compared to controls.^[53] It also aids recovery in neuromuscular disease models, accelerating torque restoration in eccentric contraction-injured mouse muscles by 7 days post-injury at 50 mg/kg, with reduced histological damage markers.^[60] Research as of 2024 emphasizes 20-hydroxyecdysone's role in coupling catabolism and anabolism to enhance biosynthetic capacity, acting as a calorie restriction mimetic that activates AMPK to balance nutrient sensing and promote longevity-like effects in mammalian models.^[51] It also shows respiratory benefits, as demonstrated in a 2024 phase 2/3 trial (COVA) where oral BIO101 (20E, 200 mg twice daily) reduced the risk of death or respiratory failure by 43.8% in adults with severe COVID-19 (n=218).^[61]

Human applications

Use as a research tool

20-Hydroxyecdysone (20E), identified in the 1960s through bioassays on insect pupae and extracts from arthropods like the crayfish Jasus lalandii, has been instrumental in elucidating insect endocrinology, particularly the hormonal control of molting and metamorphosis.^[62] Early isolation efforts, building on the 1954 discovery of ecdysone by Butenandt and Karlson, confirmed 20E as the principal active molting hormone via its ability to induce developmental transitions in ligated insect preparations, such as the blowfly Calliphora erythrocephala.^[63] This historical application in insect assays paved the way for its widespread use as a tool to decode ecdysteroid signaling pathways.^[64] In arthropod research, 20E serves as a key inducer for studying molting and developmental processes in cell cultures and mutant models. For instance, supplementation with 20E triggers differentiation and molting hormone responses in insect cell lines like Sf9 from Spodoptera frugiperda, allowing researchers to dissect cellular mechanisms of ecdysis without whole-organism variability.^[65] In genetic studies, injection or feeding of 20E rescues developmental arrest in ecdysone-deficient mutants of Drosophila melanogaster, such as l(3)dt, enabling analysis of EcR (ecdysone receptor) signaling cascades and downstream gene expression changes during metamorphosis.^[66] These applications have been crucial for mapping the hierarchical regulation of molting genes, including the identification of pulse-dependent activation of early puff genes in polytene chromosomes.^[37] In mammalian studies, 20E acts as a probe for investigating non-androgenic anabolic pathways, particularly through interactions with estrogen receptor beta (ERβ) and the mechanistic target of rapamycin (mTOR). Binding assays and hypertrophy models in skeletal muscle cells demonstrate that 20E activates ERβ to promote protein synthesis and muscle growth without androgen receptor agonism, serving as a model for selective anabolic agents.^[67] Similarly, in vivo administration in rats reveals 20E's modulation of mTORC1 signaling in liver and muscle, influencing metabolic adaptations like enhanced glucose uptake, which aids in studying insulin-independent anabolic effects. Methodologically, radiolabeled 20E, such as tritiated forms, facilitates receptor binding assays to quantify affinity and specificity in EcR complexes from arthropod tissues, providing quantitative insights into ligand-receptor dynamics. Advanced tools like CRISPR-Cas9 screening target ecdysone pathway components, such as in Bombyx mori, to identify novel regulators of 20E signal transduction and validate pathway interactions genome-wide.^[68] However, limitations include species-specific responses, where 20E efficacy varies across arthropod taxa due to receptor isoform differences, and the need for high-purity preparations to prevent confounding effects from contaminants or analogs in experimental setups.^[69]

As a dietary supplement

20-Hydroxyecdysone, commonly marketed as ecdysterone, is available over-the-counter as a dietary supplement derived from plant extracts such as spinach (Spinacia oleracea) or Cyanotis arachnoidea, typically in capsule or powder form with doses ranging from 50 to 200 mg per serving.^[70] These products are promoted primarily in the sports nutrition sector for enhancing muscle building, recovery, and athletic performance without the androgenic side effects associated with anabolic steroids.^[71] Supplements claim anabolic benefits through non-hormonal mechanisms, such as promoting protein synthesis and muscle hypertrophy, positioning ecdysterone as a safer alternative to synthetic steroids.^[70] Due to emerging evidence of performance-enhancing potential, the World Anti-Doping Agency (WADA) added ecdysterone to its Monitoring Program in 2020 to track prevalence and assess risks in competitive sports.^[72] Human evidence for efficacy remains mixed. A 2019 randomized controlled trial involving 46 resistance-trained men found that 200 mg/day of ecdysterone for 10 weeks significantly increased muscle mass and one-repetition maximum bench press strength compared to placebo, with no detected liver or kidney toxicity.^[73] However, an earlier 2006 study on 45 resistance-trained males reported no significant improvements in body composition or performance with ecdysterone supplementation.^[74] In the United States, ecdysterone is legally sold as a dietary supplement under the Dietary Supplement Health and Education Act (DSHEA), though it lacks FDA approval for specific health claims; in the European Union, it is permitted in food supplements without novel food authorization requirements for traditional botanical sources.^[71]^[75] Safety profiles indicate general tolerability, with a Phase 1 trial in 2023 showing no serious adverse events at doses up to 450 mg twice daily (900 mg/day) in healthy adults.^[76] Nonetheless, potential interactions with estrogen receptors raise concerns for hormonal modulation, particularly in women or those with endocrine conditions.^[71] By 2024-2025, ecdysterone supplements have surged in popularity within sports nutrition, driven by WADA scrutiny and social media endorsements. Quality control challenges persist, including counterfeiting in plant-based extracts and variability in ecdysteroid content, underscoring the need for third-party testing.^[77]

Potential in drug development

20-Hydroxyecdysone (20E) has emerged as a promising candidate in pharmaceutical research due to its anabolic and protective effects in mammals, particularly through activation of the Mas receptor in the renin-angiotensin-aldosterone system (RAAS). Preclinical studies have demonstrated its potential to enhance muscle protein synthesis and metabolic regulation without androgenic side effects, positioning it for development as an investigational drug under the name BIO101 by Biophytis.^[78]^[79] In neuromuscular disorders, 20E shows therapeutic promise, notably in Duchenne muscular dystrophy (DMD), where it activates the mechanistic target of rapamycin complex 1 (mTORC1) pathway to promote skeletal muscle hypertrophy and function. A 2025 preclinical study in DMD mouse models combined 20E with antisense oligonucleotide therapy targeting exon 23 of the Dmd gene, resulting in reduced dystrophic pathology, improved muscle force, and enhanced respiratory parameters. For cardio-metabolic conditions, 20E improves insulin sensitivity and glucose homeostasis; in ovariectomized rats on a high-fat-high-fructose diet, oral administration at 10 mg/kg/day for 4 weeks increased insulin receptor expression and reduced hyperglycemia and dyslipidemia. In respiratory diseases such as chronic obstructive pulmonary disease (COPD), 20E's activation of the protective RAAS arm may mitigate inflammation and muscle wasting, with preclinical evidence supporting its role in preserving respiratory muscle function in DMD models that overlap with COPD pathophysiology.^[80]^[81]^[82] Clinical trials have advanced 20E toward therapeutic use, with Phase I studies confirming safety and pharmacokinetics in healthy volunteers at doses up to 1400 mg single oral administration, showing low but measurable bioavailability. A Phase 2/3 trial (COVA) in 2024 for severe COVID-19 respiratory failure demonstrated that oral BIO101 at 12 mg/kg/day reduced the risk of death or respiratory failure by 43.8% (primary endpoint) compared to placebo, highlighting efficacy in acute respiratory conditions relevant to COPD. For metabolic syndrome, preclinical studies in gerbil models and early human data from 2020s trials indicate benefits at similar doses, including improved lipid profiles and insulin sensitivity without adverse events. Formulations such as cyclodextrin complexes have been developed to enhance solubility and absorption, addressing limitations in pharmaceutical delivery.^[2]^[83]^[84] Key challenges in 20E drug development include its poor oral bioavailability, estimated at approximately 10-12% in rodent and gerbil models due to rapid metabolism and low solubility (0.084 mg/mL in water), necessitating high doses or improved delivery systems. Efforts to develop analogs for enhanced potency are ongoing, though recent patents focus more on extraction and purification methods from plant sources like Cyanotis arachnoidea rather than synthetic derivatives.^[84]^[85]^[86] Recent advances from 2024-2025 include preclinical targeting of amyotrophic lateral sclerosis (ALS) through mTORC1 modulation for neuroprotection, with human data pending; in January 2025, Biophytis entered a co-development agreement with AskHelpU to assess BIO101's efficacy in ALS in China, and cardioprotective effects in ischemia/reperfusion injury models, partially alleviating skeletal muscle damage post-tourniquet application in mice. Patents on pharmaceutical-grade 20E extracts and RAAS-modulating formulations have supported progression, with Biophytis securing U.S. patents extending to 2039. Regulatory progress includes investigational new drug (IND) approval from the FDA in July 2024 for a Phase II trial (OBA) in obesity, and European Medicines Agency orphan drug designation in 2018 for DMD treatment.^[87]^[88]^[89]^[90]

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Dec 10, 2024 · The synthesis pathway of 20-hydroxyecdysone (20E) in arthropods is a complex multi-step biosynthetic process that begins with cholesterol and ...
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Signaling Pathways That Regulate the Crustacean Molting Gland
A pair of Y-organs (YOs) are the molting glands of decapod crustaceans. They synthesize and secrete steroid molting hormones (ecdysteroids)
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Prothoracicotropic hormone (PTTH) is a brain neuropeptide that stimulates the prothoracic glands to synthesize ecdysone, an event that leads to insect ...
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Efficient extraction and sensitive HPLC-MS/MS quantification of ...
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Detection and identification of 20-hydroxyecdysone metabolites in ...
The major 20E metabolite was 14-deoxy-20-hydroxyecdysone, which was detected in samples four to five hours after oral administration, whereas 20E was still ...<|control11|><|separator|>
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The Distribution of Phytoecdysteroids among Terrestrial Vascular ...
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Ecdysone Control of Developmental Transitions: Lessons from ...
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Ecdysone Receptor Agonism Leading to Lethal Molting Disruption in ...
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Discrete Pulses of Molting Hormone, 20-Hydroxyecdysone, During ...
Periodic pulses of the insect steroid molting hormone 20-hydroxyecdysone (20E), acting via its nuclear receptor complex (EcR/USP), control gene expression.
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Review Crustacean ecdysteriods in reproduction and embryogenesis
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The Effects of Exogenous 20-Hydroxyecdysone on the Feeding ...
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Concentration-Dependent Feeding Deterrence to 20 ... - MDPI
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A Novel Defense Against Plant-Parasitic Nematodes
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Phytoecdysteroids: a novel defense against plant-parasitic nematodes
The phytoecdysteroid, 20-hydroxyecdysone (20E), is a major molting hormone of invertebrates, possibly including nematodes. As 20E is inducible in spinach, ...Missing: annelids | Show results with:annelids<|control11|><|separator|>
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Reconstitute the 20-hydroxyecdysone Standard vial #A06120 with 1 mL of UltraPure water. Allow it to stand 5 minutes until completely dissolved and then mix ...Missing: solubility | Show results with:solubility
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In 1954, the first “moulting hormone” was isolated and named ecdysone (Butenandt and Karlson, 1954), followed by identification of the second hormone, 20- ...
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The steroid 20-hydroxyecdysone (20E) is the primary regulatory hormone that mediates developmental transitions in insects and other arthropods.
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Developmental studies on two ecdysone deficient mutants ...
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The current review investigates the beneficial biological effects of ecdysterone and turkesterone in nutrition, highlighting their roles not only in enhancing ...
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Ecdysterone Supplement Charting Growth Trajectories: Analysis ...
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Evaluating the effect of 20-hydroxyecdysone (20HE) on mechanistic ...
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Combined 20-Hydroxyecdysone and Antisense-Mediated Exon ...
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20-Hydroxyecdysone bioavailability and pharmacokinetics in ...
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[PDF] 20-Hydroxyecdysone's Cyclodextrin Complex: A Novel Anabolic ...
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A Commercial Extract of Cyanotis arachnoidea Roots as a Source of ...
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20-Hydroxyecdysone Partially Alleviates Ischemia/Reperfusion ...
Aug 7, 2025 · Duchenne muscular dystrophy is caused by loss of the dystrophin protein. This pathology is accompanied by mitochondrial dysfunction contributing ...
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[PDF] Dietary Phytoecdysteroids - Biophytis
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Aug 21, 2018 · On 27 June 2018, orphan designation (EU/3/18/2030) was granted by the European Commission to Biophytis, France, for 20-hydroxyecdysone for treatment of ...<|control11|><|separator|>