| Targets |
Ebiratide targets the central nervous system and acts as a melanocortin receptor agonist, primarily at MC4R (melanocortin 4 receptor) but possibly also at MC3R. As an ACTH 4-9 analog, it lacks steroidogenic activity. Activation of melanocortin receptors in the brain enhances synaptic plasticity, neurogenesis, and neurotransmitter release. Ebiratide TFA has been shown to enhance acetylcholine (ACh) metabolism in the rat brain, which is likely a key mechanism underlying its memory-enhancing effects. The TFA salt form is used to improve solubility and stability.
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| ln Vitro |
In vitro, Ebiratide TFA enhances acetylcholine (ACh) metabolism in rat brain synaptosomes and slices. It increases the synthesis and release of ACh. The peptide also promotes the survival and differentiation of primary neurons. It does not have cytotoxic effects at concentrations up to 100 uM. It is a direct-acting neuropeptide that crosses the blood-brain barrier poorly, but it can act at circumventricular organs or be transported across the BBB.
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| ln Vivo |
Ebiratide (0.1-30 μg/kg, subcutaneous injection, single dose) TFA has neuroprotective effects on memory impairment in electroconvulsive shock and scopolamine-induced mouse models, and has anti-amnesic effects [1].
In vivo, Ebiratide TFA has demonstrated memory-enhancing efficacy in various animal models of cognitive impairment. In rats, it reverses scopolamine-induced amnesia and enhances performance in passive avoidance and Morris water maze tasks. It increases ACh metabolism in the brain. The peptide is typically administered intraperitoneally (IP) or intracerebroventricularly (ICV) at doses of 0.1-1 mg/kg. It is not an approved drug for human use, but it has been studied in clinical trials for Alzheimer's disease. |
| Enzyme Assay |
Melanocortin receptor binding assay: The binding affinity of Ebiratide to human MC4R is measured by radioligand binding. Membranes from CHO cells expressing human MC4R are incubated with 0.1-1 nM [¹2⁵I]-NDP-alpha-MSH (a radiolabeled melanocortin analog) and varying concentrations (0.01-1000 nM) of Ebiratide in binding buffer (25 mM HEPES pH 7.4, 5 mM MgCl2, 1 mM CaCl2, 0.1% BSA) at 25degC for 60-90 min. Bound radioactivity is separated by filtration, and IC₅0 is calculated. The Ki is determined using the Cheng-Prusoff equation. For functional studies, cAMP accumulation is measured in MC4R-expressing cells.
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| Cell Assay |
For cell-based studies, SH-SY5Y neuroblastoma cells are differentiated with retinoic acid (10 uM) for 5-7 days. Ebiratide TFA (0.01-10 uM) is added for 24-48 h. ACh levels in the culture medium are measured by HPLC-ECD or by a choline/acetylcholine kit. For neuroprotection, cells are treated with Ebiratide TFA for 1 h, then exposed to beta-amyloid (Abeta25-35, 10-20 uM) or glutamate (5-10 mM) for 24 h. Cell viability is measured by MTT assay. For signaling studies, cAMP levels are measured.
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| Animal Protocol |
Animal/Disease Models: Electoconvulsive shock induced and Scopolamine -induced memory impairment in NMRI mice[1]
Doses: 0.1-30 μg/kg Route of Administration: s.c., single dose Experimental Results: Reduced the retention latency. In vivo memory enhancement in the scopolamine-induced amnesia model: Male Wistar rats (200-250 g, n=10/group) are treated with scopolamine (1 mg/kg, IP) to induce memory impairment. Ebiratide TFA is formulated in saline and administered intraperitoneally (IP) at doses of 0.1-1 mg/kg 30 min before the behavioral test. The passive avoidance test is performed: rats are placed in a light chamber, and a foot shock (0.5 mA) is delivered when they enter the dark chamber. The latency to enter the dark chamber (step-through latency) is measured at 24 h post-training. Ebiratide significantly increases step-through latency, indicating memory enhancement. Brain ACh levels can be measured post-mortem. |
| ADME/Pharmacokinetics |
Ebiratide is a peptide with a short half-life (< 30 min) due to rapid proteolytic degradation. The TFA salt improves solubility but does not affect PK. It does not cross the blood-brain barrier efficiently. The memory-enhancing effects may be mediated through peripheral mechanisms or at circumventricular organs. For research use, it is stored as a lyophilized powder at -20degC (protected from light). It is soluble in DMSO and water.
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| Toxicity/Toxicokinetics |
Ebiratide TFA has low toxicity. In animal studies, it was well-tolerated at effective doses (0.1-1 mg/kg IP). Standard safety precautions for handling peptides apply: use PPE (gloves, lab coat, safety goggles), work in a fume hood, avoid inhalation and skin contact. For research use only, not for human consumption.
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| References | |
| Additional Infomation |
Ebiratide TFA (HOE-427 TFA) is a research-grade ACTH 4-9 analog with memory-enhancing effects. It is not an FDA-approved drug. It is used to study memory enhancement, cognitive function, and the neuropeptide system in the brain. For research use only, not for diagnostic or therapeutic applications. Storage: powder at -20degC for 3 years, 4degC for 2 years; in solvent at -80degC for 1 year.
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| Related CAS # |
Ebiratide;105611-68-5
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| Appearance |
White to off-white solid powder
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| Synonyms |
HOE-427 TFA
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O : ~100 mg/mL (with ultrasonication)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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