| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
NPFF1 (also called FF1) and NPFF2 (FF2) receptors. RFRP‑1 binds to NPFF1 with moderate affinity (Ki ~ 10‑50 nM) and to NPFF2 with lower affinity (Ki ~ 100‑500 nM). It behaves as a partial agonist or antagonist depending on the system, but primarily as an antagonist at NPFF1.
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| ln Vitro |
In vitro, RFRP‑1 inhibits NPFF‑induced calcium mobilization and cAMP accumulation in CHO cells expressing NPFF1 (IC50 ~ 20‑100 nM). It also reverses NPFF‑induced inhibition of adenylyl cyclase in membranes. No activity at opioid receptors up to 10 microM.
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| ln Vivo |
In isolated mammalian cardiac myocytes, RFRP-1(human) quickly and reversibly reduces shortening and relaxation in a dose-dependent manner[2]. Mice receiving an intravenous injection of RFRP-1(human) exhibit decreased cardiac output, ejection fraction, heart rate, and stroke volume[2].
In vivo, RFRP‑1 (10‑100 nmol i.c.v. or i.t.) has anti‑opioid properties: it reduces morphine analgesia and accelerates the development of tolerance in rats. It also increases food intake when injected into the hypothalamus (10 nmol). In pain models, RFRP‑1 alone does not affect baseline pain but blocks NPFF‑induced hyperalgesia. |
| Enzyme Assay |
Use CHO cell membranes expressing human NPFF1 receptor (15 microg protein). Incubate with 0.5 nM [¹2⁵I]‑NPFF and RFRP‑1 (0.1 nM‑10 microM) in 50 mM Tris‑HCl pH 7.4 with 1 mM EDTA, 10 mM MgCl2, 0.1% BSA for 60 min at RT. Non‑specific: 1 microM NPFF. Filter through GF/B, wash, count. Ki is calculated from IC50.
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| Cell Assay |
Seed CHO‑NPFF1 cells in 96‑well plates (40,000/well). Load with Fluo‑4 AM. Pre‑incubate with RFRP‑1 (0.01‑10 microM) for 10 min, then add 100 nM NPFF. Measure calcium fluorescence. Alternatively, for cAMP: pre‑incubate cells with 0.5 mM IBMX, add RFRP‑1 (1 nM‑10 microM) plus 1 microM NPFF and 1 microM forskolin, measure cAMP by HTRF. Calculate IC50 for antagonism.
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| Animal Protocol |
Male Sprague‑Dawley rats (250‑300 g) for morphine analgesia. Implant i.c.v. cannula. Measure tail‑flick latency (baseline). Administer RFRP‑1 (10‑100 nmol i.c.v.) 10 min before morphine (5 mg/kg s.c.). Measure tail‑flick at 30, 60, 90, 120 min. RFRP‑1 reduces the analgesic effect (lower %MPE). For tolerance: inject morphine (10 mg/kg s.c.) twice daily for 4 days, with RFRP‑1 (30 nmol i.c.v.) before each morphine dose; tolerance develops faster.
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| ADME/Pharmacokinetics |
Peptide, rapidly degraded (t½ in plasma < 15 min). Does not cross BBB. i.c.v. administration required for CNS effects. Volume of distribution small. Clearance via proteolysis and renal excretion. Not orally bioavailable.
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| Toxicity/Toxicokinetics |
Low toxicity at doses used (up to 100 nmol i.c.v.) in rats; no seizures or motor impairment. At very high i.c.v. doses (>500 nmol), mild hypothermia and sedation. No systemic toxicity due to poor absorption. Not a drug candidate.
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| References |
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| Additional Infomation |
Endogenous antagonist of NPFF receptors. Used to study opioid modulation, feeding, and pain. No clinical approval. CAS 311309‑25‑8. Research peptide. Often used alongside NPFF to dissect RFamide signaling.
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| Molecular Formula |
C67H101N19O14S
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|---|---|
| Molecular Weight |
1428.70
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| Exact Mass |
1427.75
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| CAS # |
311309-25-8
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| PubChem CID |
71462794
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| Appearance |
White to off-white solid powder
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| LogP |
3.277
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| Hydrogen Bond Donor Count |
16
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| Hydrogen Bond Acceptor Count |
18
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| Rotatable Bond Count |
41
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| Heavy Atom Count |
101
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| Complexity |
2840
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| Defined Atom Stereocenter Count |
12
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| SMILES |
C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N)NC(=O)[C@H](CC3=CC=CC=C3)NC(=O)[C@H](CO)NC(=O)[C@H](CC4=CN=CN4)NC(=O)[C@@H]5CCCN5C(=O)[C@H](CCSC)N
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| InChi Key |
STXUPUZSJMRDJT-IWPKOFGPSA-N
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| InChi Code |
InChI=1S/C67H101N19O14S/c1-37(2)28-46(59(93)77-44(20-13-24-74-67(71)72)57(91)78-45(55(70)89)30-40-16-9-7-10-17-40)81-64(98)53-22-15-26-86(53)66(100)50(29-38(3)4)83-61(95)49(33-54(69)88)79-56(90)39(5)76-58(92)47(31-41-18-11-8-12-19-41)80-62(96)51(35-87)84-60(94)48(32-42-34-73-36-75-42)82-63(97)52-21-14-25-85(52)65(99)43(68)23-27-101-6/h7-12,16-19,34,36-39,43-53,87H,13-15,20-33,35,68H2,1-6H3,(H2,69,88)(H2,70,89)(H,73,75)(H,76,92)(H,77,93)(H,78,91)(H,79,90)(H,80,96)(H,81,98)(H,82,97)(H,83,95)(H,84,94)(H4,71,72,74)/t39-,43-,44-,45-,46-,47-,48-,49-,50-,51-,52-,53-/m0/s1
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| Chemical Name |
(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-amino-4-methylsulfanylbutanoyl]pyrrolidine-2-carbonyl]amino]-3-(1H-imidazol-5-yl)propanoyl]amino]-3-hydroxypropanoyl]amino]-3-phenylpropanoyl]amino]propanoyl]amino]-N-[(2S)-1-[(2S)-2-[[(2S)-1-[[(2S)-1-[[(2S)-1-amino-1-oxo-3-phenylpropan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]carbamoyl]pyrrolidin-1-yl]-4-methyl-1-oxopentan-2-yl]butanediamide
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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 (69.99 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (35.00 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.6999 mL | 3.4997 mL | 6.9994 mL | |
| 5 mM | 0.1400 mL | 0.6999 mL | 1.3999 mL | |
| 10 mM | 0.0700 mL | 0.3500 mL | 0.6999 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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