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RF9

Alias: RF-9 RF9 RF 9
Cat No.:V7579 Purity: ≥98%
RF9 is a potent and specific neuropeptide FF receptor blocker (antagonist) with Kis of 58 and 7 nM for hNPFF1R and hNPFF2R respectively.
RF9
RF9 Chemical Structure CAS No.: 876310-60-0
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
Size Price Stock Qty
1mg
5mg
Other Sizes

Other Forms of RF9:

  • RF9 hydrochloride
Official Supplier of:
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Top Publications Citing lnvivochem Products
Product Description
RF9 is a potent and specific neuropeptide FF receptor blocker (antagonist) with Kis of 58 and 7 nM for hNPFF1R and hNPFF2R respectively.
Biological Activity I Assay Protocols (From Reference)
ln Vitro
In Neuro 2A cells, RF9 (10 μM) pretreatment totally inhibited NPFF-induced neurite development [2].
ln Vivo
Heroin-induced delayed hyperalgesia and tolerance are avoided when RF9 (0.1 mg/kg, subcutaneous injection) is administered concurrently with heroin [1]. MAP and heart rate did not alter significantly after receiving RF9 (10 μg) infusion alone. On the other hand, the NPFF-induced increases in heart rate and MAP are dramatically inhibited when RF9 is delivered in conjunction with NPFF [1].
Animal Protocol
Animal/Disease Models: Rat[1].
Doses: 0.1 mg/kg.
Route of Administration: 0.3 mg/kg heroin or saline was injected subcutaneously (sc) (sc) 30 minutes before basal nociceptive threshold in rats.
Experimental Results: Against delayed heroin-induced hyperalgesia and related tolerance.
References

[1]. RF9, a potent and selective neuropeptide FF receptor antagonist, prevents opioid-induced tolerance associated with hyperalgesia. Proc Natl Acad Sci U S A. 2006 Jan 10;103(2):466-71.

[2]. Discovery of Two Novel Branched Peptidomimetics Containing endomorphin-2 and RF9 Pharmacophores: Synthesis and Neuropharmacological Evaluation. Bioorg Med Chem. 2019 Feb 15;27(4):630-643.

Additional Infomation
RF9 is a dipeptide formed from L-arginine and L-phenylalaninamide residues in which a hydrogen attached to the nitrogen of the alpha-amino group of the arginine residue has been replaced by a 1-adamantanecarbonyl group. It has been reported to be a potent and selective antagonist of neuropeptide FF (NPFF) receptors, but more recently found to be an agonist at both NPFF1R and the kisspeptin receptor (KISS1R). It has a role as a neuropeptide FF receptor antagonist, a neuropeptide FF receptor agonist and a kisspeptin receptor agonist. It is functionally related to an Arg-Ala.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C26H38N6O3
Molecular Weight
482.63
Exact Mass
482.3
CAS #
876310-60-0
Related CAS #
RF9 hydrochloride
PubChem CID
53320361
Appearance
White to off-white solid powder
LogP
4.325
Hydrogen Bond Donor Count
5
Hydrogen Bond Acceptor Count
4
Rotatable Bond Count
11
Heavy Atom Count
35
Complexity
777
Defined Atom Stereocenter Count
2
SMILES
C1C2CC3CC1CC(C2)(C3)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC4=CC=CC=C4)C(=O)N
InChi Key
UMKHUSRDQFQHAK-RNJMTYCLSA-N
InChi Code
InChI=1S/C26H38N6O3/c27-22(33)21(12-16-5-2-1-3-6-16)31-23(34)20(7-4-8-30-25(28)29)32-24(35)26-13-17-9-18(14-26)11-19(10-17)15-26/h1-3,5-6,17-21H,4,7-15H2,(H2,27,33)(H,31,34)(H,32,35)(H4,28,29,30)/t17?,18?,19?,20-,21-,26?/m0/s1
Chemical Name
N-[(2S)-1-[[(2S)-1-amino-1-oxo-3-phenylpropan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]adamantane-1-carboxamide
Synonyms
RF-9 RF9 RF 9
HS Tariff Code
2934.99.9001
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)
Solubility Data
Solubility (In Vitro)
DMSO : ~125 mg/mL (~259.00 mM)
H2O : ~16.67 mg/mL (~34.54 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.08 mg/mL (4.31 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 (4.31 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.

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Solubility in Formulation 3: ≥ 2.08 mg/mL (4.31 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 corn oil and mix evenly.


Solubility in Formulation 4: 10 mg/mL (20.72 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication (<60°C).

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 2.0720 mL 10.3599 mL 20.7198 mL
5 mM 0.4144 mL 2.0720 mL 4.1440 mL
10 mM 0.2072 mL 1.0360 mL 2.0720 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.

Calculator

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What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
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What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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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.

Biological Data
  • Screening of RFamide derivatives on hNPFF2R. (A) hNPFF2R membranes were labeled with [125I]Tyr-NPFF, and three concentrations of RFamide derivatives were tested in competition experiments. Each concentration was tested in duplicate. Results for the reference and the six most active compounds are shown. Arrowheads indicate compounds that were selected for further characterization. (B) Structures of RF2, RF9, RF48, RF49, and BIBP3226.[1].Simonin F, et al. RF9, a potent and selective neuropeptide FF receptor antagonist, prevents opioid-induced tolerance associated with hyperalgesia. Proc Natl Acad Sci U S A. 2006 Jan 10;103(2):466-71.
  • In vitro characterization of compounds selected from the screening. (A) Stimulation of [35S]GTPγS binding to hNPFF2R by NPFF (▪), RF2 (▴), RF48 (▾) and RF49 (♦). (B) Stimulation of [35S]GTPγS binding to hNPFF2R by NPFF alone (▪) or NPFF in presence of 7.5 μM RF9 (▴). RF9 shifted the concentration-effect curve of NPFF to the right by ≈160-fold. Data are expressed as percentage of basal [35S]GTPγS binding and represent mean ± SE from at least two separate experiments in triplicate. (C) RF9 (black bars) reversed the inhibition of forskolin-induced cAMP by NPVF in CHO-hNPFF1R cells. RF9 alone was inactive, whereas NPVF (white bar) inhibited ≈60% of the stimulated cAMP. Error bars represent the mean ± SEM of data from three experiments performed in duplicate.[1].Simonin F, et al. RF9, a potent and selective neuropeptide FF receptor antagonist, prevents opioid-induced tolerance associated with hyperalgesia. Proc Natl Acad Sci U S A. 2006 Jan 10;103(2):466-71.
  • RF9 blocks blood pressure effects of NPFF. (A) Changes in MAP [expressed in mmHg (1 mmHg = 133 Pa)] in rats receiving either i.c.v. saline (♦), NPFF (▪), RF9 (×), or NPFF and RF9 applied together (▴). Time 0 indicates the injection point for i.c.v. saline or drug applications. (B) Heart rate changes that accompany MAP alterations in A. Pooled MAP and heart rate data are from five animals. *, significant difference in MAP or heart rate compared with control (P < 0.05).[1].Simonin F, et al. RF9, a potent and selective neuropeptide FF receptor antagonist, prevents opioid-induced tolerance associated with hyperalgesia. Proc Natl Acad Sci U S A. 2006 Jan 10;103(2):466-71.
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