| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg | |||
| Other Sizes |
| Targets |
hNPFF1R 58 nM (Ki) hNPFF2R 75 nM (Ki)
RF9 hydrochloride targets the neuropeptide FF (NPFF) receptors, specifically the hNPFF1R and hNPFF2R subtypes. It acts as a selective antagonist, blocking the binding of endogenous NPFF ligands. The Ki values are 58 nM for hNPFF1R and 7 nM for hNPFF2R. NPFF receptors are involved in pain modulation, opioid tolerance, and cardiovascular regulation. |
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| ln Vitro |
RF9 (10 μM) partially prevents Neuro 2A cells' neurite outgrowth that is caused by NPFF[2].
In vitro, RF9 at 10 microM partially prevents Neuro 2A cells' neurite outgrowth induced by NPFF. This confirms its activity as an NPFF receptor antagonist. The compound is highly selective and potent, with sub-nanomolar to low nanomolar affinity for NPFF receptors. It is used to study NPFF-mediated signaling pathways, including calcium mobilization and MAPK activation. |
| ln Vivo |
Tolerance and heroin-induced delayed hyperalgesia are avoided when RF9 (0.1 mg/kg, sc) is also administered[1]. No discernible change in heart rate or MAP occurs when RF9 (10 μg) is administered alone. In contrast, when NPFF is administered in combination with RF9, the MAP and heart rate increases elicited by NPFF are greatly inhibited[1].
In vivo, RF9 prevents opioid-induced tolerance and heroin-induced delayed hyperalgesia when administered subcutaneously at 0.1 mg/kg. It also inhibits the development of tolerance to morphine analgesia, suggesting its potential for managing opioid side effects. The compound crosses the blood-brain barrier and is active in the central nervous system. |
| Enzyme Assay |
A cell-free receptor binding assay is performed using membranes from CHO or HEK293 cells expressing human NPFF receptors. Membranes (10-20 ug protein) are incubated with a radiolabeled NPFF ligand (e.g., [3H]NPFF, 0.5-2 nM) and increasing concentrations (0.01-10,000 nM) of RF9 hydrochloride in binding buffer (50 mM Tris-HCl, pH 7.4, 5 mM MgCl2, 0.1% BSA). Nonspecific binding is determined using 10 uM NPFF. Bound radioligand is separated by filtration through GF/B filters and counted. Ki values are calculated from competition curves.
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| Cell Assay |
A functional calcium mobilization assay is performed using cells expressing NPFF receptors. HEK293 cells stably expressing hNPFF1R or hNPFF2R are seeded in 96-well plates and loaded with Fluo-4 AM (4 uM). RF9 hydrochloride (0.1-1000 nM) is added 10 min before the addition of an NPFF agonist (e.g., NPFF, 10 nM). The reduction in calcium signal compared to the agonist-only control is measured. The IC50 for antagonist activity is calculated from the inhibition curve.
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| Animal Protocol |
Animal/Disease Models: Rats[1]
Doses: 0.1 mg/kg. Route of Administration: SC 30 min before 0.3 mg/kg heroin or saline on basal nociceptive threshold in rats. Experimental Results: Opposed to delaye heroin-induced hyperalgesia and associated tolerance. RF9 hydrochloride can be studied in mouse models of opioid tolerance. Male C57BL/6 mice (20-25 g, n=8-10 per group) are injected subcutaneously with morphine (10 mg/kg) twice daily for 7 days to induce tolerance. RF9 (0.1-1 mg/kg, s.c.) is administered 15 min before each morphine injection. Antinociception is assessed by the tail-flick test (radiant heat, latency to flick tail) 30 min after morphine administration. RF9 is expected to prevent the development of morphine tolerance, maintaining antinociceptive efficacy over time. |
| ADME/Pharmacokinetics |
RF9 hydrochloride has a molecular weight of 519.08 and the molecular formula C26H39ClN6O3. The powder should be stored at -20degC, sealed, and away from moisture. In solvent, it is stable for 6 months at -80degC or 1 month at -20degC. The compound is soluble in DMSO (50 mg/mL). For in vivo use, it can be formulated in 10% DMSO, 40% PEG300, 5% Tween 80, 45% saline.
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| Toxicity/Toxicokinetics |
No detailed toxicity data for RF9 hydrochloride are available. In animal studies, the compound was well-tolerated at doses up to 1 mg/kg (s.c.) with no overt signs of systemic toxicity. As an NPFF receptor antagonist, the primary safety concern is potential modulation of pain perception and opioid responses. Standard safety precautions for handling research chemicals should be followed.
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| References |
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| Additional Infomation |
RF9 hydrochloride is a research-grade compound and is not approved for clinical use. It is a potent and selective NPFF receptor antagonist used to study pain modulation, opioid tolerance, and other NPFF-mediated processes. This product is for research use only and not for human therapeutic applications. Store as a powder at -20degC.
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| Molecular Formula |
C26H39CLN6O3
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|---|---|
| Molecular Weight |
519.08
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| Related CAS # |
RF9;876310-60-0
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| Appearance |
White to off-white solid powder
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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) |
DMSO :~50 mg/mL (~96.32 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.01 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (4.01 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (4.01 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9265 mL | 9.6324 mL | 19.2649 mL | |
| 5 mM | 0.3853 mL | 1.9265 mL | 3.8530 mL | |
| 10 mM | 0.1926 mL | 0.9632 mL | 1.9265 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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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