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Testolone (RAD140 liquid) is a selective androgen receptor modulator (SARM), originally developed by Radius Health, for the treatment of muscle atrophy and osteoporosis. As a non-steroidal compound, RAD140 selectively activates androgen receptors in skeletal muscles and neural tissues, while reducing side effects on tissues such as the prostate, skin, and hair follicles. Its anabolic effect is similar to that of synthetic steroids, but with higher receptor selectivity and potentially better safety.RAD140 liquid is the liquid oral formulation of this compound. It is commonly used by fitness enthusiasts or athletes to enhance muscle mass and strength. Studies have shown that it can significantly increase lean body mass while avoiding the estrogen or progesterone-related side effects (such as gynecomastia) associated with traditional steroids.
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Testolone (RAD140 liquid) COA
Core Applications in the Scientific Research Field
Testolone (RAD140 liquid) is a commonly used research tool in scientific research, primarily for investigating the mechanism of action of the androgen receptor and its associated physiological functions. As a high-purity preparation, it is usually suspended in PEG-400 to ensure solubility and concentration stability. Each batch is tested by a third-party ISO-certified laboratory to verify its purity, concentration and identity, thus guaranteeing the accuracy and reliability of scientific research data.
Researchers mainly utilize its tissue selectivity to conduct preclinical experiments on muscle growth regulation, bone metabolism, neuroprotection and other aspects-for example, exploring its effects on the proliferation and differentiation of skeletal muscle cells and its protective role in neurodegenerative disease models, which provides a theoretical basis for drug development for related diseases. In addition, its liquid dosage form allows for precise control of the administration dose, adapting to the dosing requirements of different animal models, and is widely used in relevant studies on experimental animals such as rats and mice.
Applications in Physical Performance Enhancement and Fitness-Related Research
In the field of physical performance enhancement, RAD140 Liquid has become a focus of relevant research due to its potent anabolic activity. Studies have shown that it can effectively stimulate muscle growth and enhance muscle strength, while potentially assisting in reducing body fat by accelerating metabolism, with significantly milder side effects compared to traditional steroids. Its liquid dosage form features a faster absorption rate: it takes effect 15–30 minutes after sublingual administration, and its bioavailability is notably higher than that of tablets, capsules and other dosage forms, enabling it to rapidly reach an effective blood drug concentration.
Scientific research has found that a daily dose of 5–20mg can exhibit obvious effects of muscle growth and strength enhancement, with 10mg considered a relatively optimal range. Moreover, due to its long half-life of 60 hours, a single daily administration is sufficient to maintain a stable therapeutic effect. In addition, it can promote erythropoiesis, increase blood supply to muscle tissue, accelerate post-exercise muscle recovery and reduce the risk of sports injuries, serving as an important tool for scientific research related to physical training.
Explorations of Potential Applications in the Medical Field
Although clinical data is currently limited, the potential application prospects of RAD140 Liquid in the medical field have been initially explored. In terms of muscle wasting-related diseases, it can effectively inhibit muscle loss and is applicable to preclinical studies on diseases such as sarcopenia and cachexia, providing new ideas for muscle protection in such patients.
In the field of bone health, it can specifically bind to androgen receptors in bone tissue, promote osteoblast activity and inhibit osteoclast proliferation, showing promise for research and potential treatment of osteoporosis-especially for the maintenance of bone health in elderly men or postmenopausal women. Furthermore, the latest preclinical studies have found that RAD140 Liquid possesses neuroprotective effects: in hippocampal neuron cultures and kainate-induced injury rat models, it can reduce neuronal death and exert a protective effect by activating the MAPK signaling pathway, which provides a new direction for the research of neurodegenerative diseases such as Alzheimer's disease.
Auxiliary Applications in Special Scenarios and Precautions
The flexible dose adjustment of RAD140 Liquid makes it suitable for the needs of different research scenarios. The dose can be precisely adjusted with a dropper or syringe according to the experimental purpose or the body weight and physical condition of the research subject, overcoming the limitation of fixed doses in tablet formulations. However, it should be noted that it has a certain testosterone-suppressive effect: preclinical studies have shown that after 28 consecutive days of administration, the testosterone levels of experimental animals are suppressed to 200–300ng/dl, and the higher the dose, the more obvious the suppressive effect may be, which is also an indicator that requires key monitoring in scientific research.
At the same time, safety data for its use in females is extremely limited, and it may affect the level of free testosterone in the body, so it is not recommended for human studies involving females for the time being. In addition, as a research chemical, it must be hermetically stored in a cool, dry and light-protected environment to avoid drug degradation caused by high temperature and humidity, which would affect its use effect.
However, its long-term safety has not been fully verified. Common side effects include mild headaches, fatigue, and testosterone suppression. Due to the lack of clinical approval, RAD140 is currently only available as a research chemical for circulation. The World Anti-Doping Agency (WADA) has listed it as a prohibited substance.
Fundamentals of Chemistry and Synthetic Processes
Chemical Basis
Testolone (RAD140 liquid) is a non-steroidal selective androgen receptor modulator (SARM), with the molecular formula of C₂₀H₁₆ClN₅O₂ and a molecular weight of 393.826 g/mol. Its chemical structure contains the following key features:
Oxazole ring
As the core framework, this ring structure enhances the rigidity of the molecule through conjugation effects, and serves as the binding site for androgen receptors, endowing it with the ability to selectively activate.
Cyano group and chlorine atom
The cyano group (-CN) acts as a strong electron-withdrawing group and may participate in photolysis reactions through an electron transfer mechanism, but in synthesis, it is used as a key functional group for guiding subsequent reactions. The chlorine atom (Cl) stabilizes the molecular electron cloud distribution through an inducing effect.
Hydroxypropyl side chain
Provides a polar group, enhances intermolecular hydrogen bonding, improves solubility (solubility in DMSO ≥ 90 mg/mL), and reduces oxidation reaction activity through spatial steric hindrance.
Synthesis Process
The synthesis of RAD140 requires the completion of multiple organic reactions. The core steps are as follows:
Raw materials: 2-chloro-4-fluoro-3-methylbenzamide and H-D-Thr-OH (hydroxythreonine).
Reaction conditions: Mix in DMSO, add potassium carbonate (K₂CO₃) as a base, and stir the reaction at 75°C for 24 hours.
Product: Compounds 9 (purity ≥ 95%) are generated through acidification, extraction, and column chromatography purification.
Raw material: Compound 9 and 4-cyanobenzhydrazine.
Reaction conditions: Mix in THF, cool to -15°C under nitrogen protection, add hydroxybenzotriazole (HOBT), triethylamine, and carbodiimide hydrochloride (EDCI) as condensing agents, and stir the reaction overnight.
Product: Yellow solid compound 10 is generated (yield 78%), purified by filtration, washing, and boiling.
Siloxane protection: React using tert-butyl dimethyl chlorosilane (TBDMSCI) and imidazole in dichloromethane to generate intermediate 10a (yield 84%).
Iodination reaction: Introduce iodine atoms through nucleophilic substitution reaction with triphenylphosphine, iodine crystals, and triethylamine, generating compound 10b (yield 84%).
De-protection: Remove the siloxane protection group using tetrabutyl fluoride ammonium (TBAF) at -40°C to obtain crude RAD140 (yield 68%), and purify it by silica gel chromatography to obtain a white solid.
Liquid formulation and stability optimization
Testolone (RAD140 liquid) needs to be preserved for a long time through solvent dissolution and the addition of stabilizers:
Solvent selection: Commonly, high-purity DMSO (≥99.9%) or polyethylene glycol 400 (PEG400) is used as the solvent to ensure complete dissolution of RAD140 (concentration 10-20 mg/mL).
Stabilizer addition: 0.1% ascorbic acid (vitamin C) or butylated hydroxyanisole (BHA) is added to inhibit oxidative degradation, and 0.01% EDTA disodium is added as a chelating agent to prevent metal ion catalytic degradation.
Storage conditions:
Avoid light: Use brown glass bottles or aluminum foil bags for packaging to prevent photolysis reactions.
Low temperature: It is recommended to freeze at -20℃ for storage. The stability at 25℃ ambient temperature can only be maintained for 1 month (content decrease ≤ 5%), while under freezing conditions, it can be extended to 2 years.
Quality Control and Detection Methods
High Performance Liquid Chromatography (HPLC)
Use a C18 column (4.6×250 mm, 5 μm), with a mobile phase of acetonitrile-water (containing 0.1% formic acid) for gradient elution, detection wavelength 254 nm, the retention time of RAD140 is approximately 8.5 minutes, purity needs to be ≥ 98%.
Liquid Chromatography-Mass Spectrometry (LC-MS)
Detected by electrospray ionization (ESI) positive ion mode, the molecular ion peak (m/z) is 394.1 ([M+H]⁺), confirming the accuracy of molecular weight.
Nuclear Magnetic Resonance (NMR)
¹H NMR (400 MHz, DMSO-d₆) shows characteristic peaks: δ 8.52(s, 1H, cyanide meta-adjacent proton), 7.98(d, 2H, benzene ring proton), 4.21(m, 1H, hydroxypropyl proton), verifying the integrity of the structure.
Liquid formulation and stability optimization
Liquid Formula Design
Solvent Selection
DMSO (Dimethyl Sulfoxide): As the core solvent, DMSO can completely dissolve RAD140 (with a concentration up to 10-20 mg/mL), but the moisture content needs to be controlled (≤0.1%) to avoid hydrolysis reactions.
PEG400 (Polyethylene Glycol 400): A mild solvent as an alternative to DMSO, suitable for users sensitive to DMSO, but with a slightly lower solubility (about 8-15 mg/mL), it needs to be dissolved through heating (40-50℃).
Mixed Solvent System: DMSO and PEG400 are mixed in a 1:1 ratio, which can balance solubility and irritation, while reducing viscosity and improving the fluidity of the droplet.
Stabilizer Addition
Antioxidant: Add 0.1% ascorbic acid (vitamin C) or butylated hydroxyanisole (BHA) to inhibit oxidation degradation and extend the shelf life to 18 months (at room temperature).
Chelating Agent: Add 0.01% EDTA disodium, chelating metal ions (such as Fe³⁺, Cu²⁺), to prevent catalytic degradation.
pH Regulator: Use phosphate buffer solution (PBS) or citrate buffer solution to control the pH within the range of 4-6, avoiding structural damage under acidic or alkaline conditions.
Osmotic Pressure Regulation
If the formula is used for injection or transdermal administration, glycerol or glucose should be added to regulate the osmotic pressure to the physiological range (280-310 mOsm/kg), reducing irritation.
Stability Optimization Strategies
Light Stability Enhancement
Shading Packaging: Use brown glass bottles or aluminum foil bags for packaging to block 99.9% of ultraviolet light (UV-A/B).
Light Stabilizer: Add 0.05% titanium dioxide (TiO₂) nanoparticles to the formula, through physical shielding and free radical capture mechanisms to reduce photolysis reactions.
Temperature Control
Freezing Storage: Recommend freezing at -20℃ for storage, with stability only maintaining for 1 month at 25℃ (content decrease ≤5%), while freezing conditions can extend to 2 years.
Thermal Stability Test: In the 40℃ accelerated test, Testolone (RAD140 liquid) is stable for 6 months in DMSO (content ≥95%), while in PEG400 it is only stable for 3 months, requiring optimization of the formula ratio.
Oxidation Degradation Inhibition
Nitrogen Protection: During filling, introduce nitrogen to exclude oxygen, reducing the risk of oxidation.
Stabilizer Synergism: Ascorbic acid combined with BHA can reduce the oxidation degradation rate by 80%, extending the shelf life to 24 months.
Quality Control and Testing Methods
High Performance Liquid Chromatography (HPLC)
Use a C18 chromatographic column (4.6×250 mm, 5 μm), with a mobile phase of acetonitrile-water (containing 0.1% formic acid) for gradient elution, with a detection wavelength of 254 nm. The retention time of RAD140 is approximately 8.5 minutes, and the purity needs to be ≥98%.
Liquid Chromatography-Mass Spectrometry (LC-MS)
Detect through electrospray ionization (ESI) positive ion mode, the molecular ion peak (m/z) is 394.1 ([M+H]⁺), confirming the accuracy of molecular weight.
Stability Acceleration Test
Place in a 40℃/75% RH environment for 6 months, take samples regularly to detect content and impurities. If the content decrease ≤10% and impurities ≤1.0%, the formula stability is qualified.
Industrial Application Optimization Case
A Biotechnology Company Formula
Composition: RAD140 10 mg/mL, DMSO 70%, PEG400 20%, ascorbic acid 0.1%, EDTA disodium 0.01%.
Stability Data: Stored at 25℃ for 12 months, the content decreased by 3.2%, impurities 0.8%; Stored at -20℃ for 24 months, the content decreased by 1.5%, impurities 0.3%.
Transdermal Patch Formula
Composition: RAD140 5 mg/g, polyacrylate pressure-sensitive adhesive 80%, propylene glycol 10%, titanium dioxide 5%.
Stability data: When stored at 40℃ for 3 months, the transdermal flux remained stable at 12.5 μg/cm²/h, and the content decreased by ≤5%.
FAQ
How long does RAD140 stay in your body?
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RAD140 and its hydroxy metabolites are almost completely conjugated with glucuronic acid. It is not clear whether these are O- or N-glucuronides. In the last urine sample RAD140 was still perfectly detectable, and therefore its detection period could be roughly estimated as 10-14 days.
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