| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| Other Sizes |
| Targets |
5-HT2B Receptor 11.2 nM (Ki) 5-HT2A Receptor 1516 nM (Ki) 5-HT2C Receptor 324 nM (Ki)
5-HT2B receptor (agonist, Ki = 11.2 nM). |
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| ln Vitro |
In rats, arteries with a significant drop in threshold (the mesenteric resistance artery and the aorta) contract when exposed to (1 nM to 100 μM) with (+)-norfenfluramine[1]. Aortic contraction is induced in tissues of normotensive and hypertensive rats by (+)-Norfenfluramine (1 and 10 μM, 3 min)[1]. Rat hippocampus synaptosomes release 5-HT in a Ca2+-dependent manner when exposed to (0–10 μM)-Norfenfluramine for three minutes [2].
In radioligand binding assays, (+)-Norfenfluramine is a selective 5-HT2B receptor agonist with a Ki of 11.2 nM. It effectively stimulates the hydrolysis of phosphoinositide (inositol phosphates, IP) and increases intracellular Ca2+ concentration through activation of Gq-coupled 5-HT2B receptors. These cellular signaling cascades are associated with fibroblast proliferation and valvular remodeling. |
| ln Vivo |
In conscious SHAM and DOCA-salt rats, (+)-Norfenfluramine (1-300 μg/kg, iv) generates a pressor response[1]. Rat telencephalon and brainstem 5-HT and 5-HIAA levels are decreased by (+)-Norfenfluramine (2.5 and 5 mg/kg, ip)[3].
The principal pharmacological activity of (+)-Norfenfluramine is 5-HT2B agonism. It was the primary mediator of the adverse cardiovascular effects (valvular heart disease and pulmonary hypertension) associated with fenfluramine use, which led to the withdrawal of fenfluramine-based anorectic drugs from the market. It is used as a tool compound to model drug-induced valvulopathy. |
| Enzyme Assay |
Standard radioligand binding assays for 5-HT2B receptors are performed using membrane preparations from CHO or HEK-293 cells stably expressing human 5-HT2B receptors. [3H]-5-HT (serotonin) or [3H]-LSD is used as the radioligand. Test compound ((+)-Norfenfluramine) is incubated with membranes at varying concentrations (10-¹¹ to 10-⁵ M) for 60 minutes at 25degC. Nonspecific binding is determined in the presence of 10 uM unlabeled 5-HT or 10 uM methysergide. Bound radioligand is separated by vacuum filtration through glass fiber filters and counted by liquid scintillation. Ki values (11.2 nM) are calculated from competition curves using the Cheng-Prusoff equation. For selectivity profiling, assays for other serotonin receptors (5-HT1A, 5-HT2A, 5-HT2C, 5-HT1B, 5-HT1D, 5-HT6) and other GPCRs are performed similarly.
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| Cell Assay |
Cell-based functional assays for 5-HT2B agonism are performed using CHO or HEK-293 cells stably expressing human 5-HT2B receptors. The 5-HT2B receptor is Gq-coupled, activating phospholipase C (PLC). For IP1 accumulation assays (a direct readout of PLC activity), cells are seeded in 96-well plates. After overnight culture, growth medium is replaced with assay buffer (HBSS containing HEPES, pH 7.4) containing LiCl (10-50 mM, to inhibit IP1 degradation). Cells are then treated with serial dilutions of (+)-Norfenfluramine (10-¹¹ to 10-⁵ M) for 30-60 minutes at 37degC. The accumulated IP1 (a stable downstream metabolite of IP3) is quantified by HTRF (Cisbio IP-One kit). For calcium flux assays, cells are loaded with Fluo-4 AM dye and fluorescence is measured upon agonist addition. EC50 values are calculated from concentration-response curves. The EC50 for (+)-Norfenfluramine at 5-HT2B is typically in the low nanomolar range (e.g., ∼10-50 nM), consistent with its high binding affinity. A standard 5-HT2B antagonist (e.g., SB204741 or RS-127445) is included as a control to confirm specificity.
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| Animal Protocol |
Animal/Disease Models: Conscious SHAM and DOCA-salt rats[1].
Doses: 1-300 μg/kg Route of Administration: intravenous (iv) injection (iv), given in a cumulative fashion at 6-min intervals. Experimental Results: Induced pressor response in conscious SHAM and DOCA-salt rats. (change in mean arterial blood pressure at 300 μg/kg, mm Hg, SHAM vehicle=36, SHAM ketanserin=7, DOCA=51, DOCA ketanserin=19). In vivo studies for (+)-Norfenfluramine are typically conducted in rodents (rats or mice) to model 5-HT2B-mediated valvulopathy and cardiac fibrosis. The compound is dissolved in saline and administered via subcutaneous (s.c.) or intraperitoneal (i.p.) injection (e.g., 1-10 mg/kg/day) for a period of 1-28 days. Animals are sacrificed, and heart tissues (specifically the valves, left ventricle) are harvested. Pathological endpoints include: (1) Histological assessment of valvular thickness (hematoxylin and eosin, Masson‘s trichrome staining) and collagen deposition; (2) Immunohistochemistry for markers of proliferation (Ki-67) and myofibroblast activation (alpha-SMA); (3) Molecular markers of fibrosis (qPCR for collagen I, collagen III, TGF-beta1, and CTGF). For a non-valvulopathy endpoint, the compound can be used in acute studies to measure changes in pulmonary artery pressure in anesthetized rats (a surrogate for pulmonary arterial hypertension). |
| ADME/Pharmacokinetics |
Metabolism / Metabolites
Known metabolites of (s)-desfenfluramine include 2-(3-(trifluoromethyl)phenyl)ethylamine. (+)-Norfenfluramine is a metabolite, not a therapeutic agent. Its pharmacokinetic properties in vivo are derived from studies of the parent drug fenfluramine. Fenfluramine is rapidly absorbed and extensively metabolized in the liver, primarily by CYP2D6 and other CYP enzymes, to norfenfluramine (the major active metabolite). (+)-Norfenfluramine has a longer half-life (∼20 hours) and higher potency than the parent drug, leading to its accumulation upon chronic dosing and causing the delayed onset of cardiotoxicity. |
| Toxicity/Toxicokinetics |
(+)-Norfenfluramine is a research tool and is not used therapeutically. Its toxicological profile is of significant interest because it is the primary mediator of the valvular heart disease associated with fenfluramine. Chronic activation of 5-HT2B receptors on heart valves stimulates proliferation of valvular interstitial cells, leading to leaflet thickening, retraction, and regurgitation. In animal models, chronic administration leads to dose- and time-dependent valvulopathy. Acute effects include mydriasis (pupil dilation), hyperthermia, and behavioral changes. Standard laboratory safety precautions must be observed.
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| References |
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| Additional Infomation |
(2S)-1-[3-(trifluoromethyl)phenyl]-2-propylamine belongs to the amphetamine class of drugs. It is an analogue of fenfluramine, inhibiting serotonin uptake and potentially promoting serotonin release. It is used as an appetite suppressant and in animal experiments. See also: norfenfluramine (note moved to).
(+)-Norfenfluramine is a key research tool for studying the mechanism of drug-induced valvular heart disease. It is the principal metabolite of the now-withdrawn obesity drug fenfluramine. Activation of 5-HT2B receptors causes mitogenic stimulation of valvular interstitial cells via the Gq-PLC-IP3-Ca2+ pathway. The compound is also used in research on pulmonary arterial hypertension (PAH). As a controlled substance analog in many jurisdictions, it is strictly for in vitro research and not for human use. |
| Molecular Formula |
C10H12F3N
|
|---|---|
| Molecular Weight |
203.20
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| Exact Mass |
203.092
|
| CAS # |
19036-73-8
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| Related CAS # |
(+)-Norfenfluramine hydrochloride;37936-89-3;Norfenfluramine;1886-26-6
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| PubChem CID |
9815618
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| Appearance |
Colorless to light yellow liquid
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| Density |
1.152g/cm3
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| Boiling Point |
215.2ºC at 760mmHg
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| Flash Point |
88.9ºC
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| Vapour Pressure |
0.15mmHg at 25°C
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| Index of Refraction |
1.467
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| LogP |
3.295
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
14
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| Complexity |
179
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| Defined Atom Stereocenter Count |
1
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| SMILES |
FC(C1=CC=CC(=C1)C[C@H](C)N)(F)F
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| InChi Key |
MLBHFBKZUPLWBD-ZETCQYMHSA-N
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| InChi Code |
InChI=1S/C10H12F3N/c1-7(14)5-8-3-2-4-9(6-8)10(11,12)13/h2-4,6-7H,5,14H2,1H3/t7-/m0/s1
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| Chemical Name |
(2S)-1-[3-(trifluoromethyl)phenyl]propan-2-amine
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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: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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: 100 mg/mL (492.13 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (12.30 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 25.0 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: ≥ 2.5 mg/mL (12.30 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. View More
Solubility in Formulation 3: 2.5 mg/mL (12.30 mM) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 4.9213 mL | 24.6063 mL | 49.2126 mL | |
| 5 mM | 0.9843 mL | 4.9213 mL | 9.8425 mL | |
| 10 mM | 0.4921 mL | 2.4606 mL | 4.9213 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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