25CN-NBOH

Protein target name:

HTR2A, HTR2C,

Mechanism of action:

Selective Agonist

Recommended Concentration for use in cells:

up to 500 nM

Probe Availability

TOCRIS

Probe Information

Target and Probe Information

Target Details

Target class:

GPCR

Subclass:

Serotonin Receptor

HUGO ID:

HGNC:5293 HGNC:5294 HGNC:5295

Entrez gene ID:

3356 3357 3358

UNIPROT ID:

P28223 P28335

Target alias:

5-HT2A, HTR2, 5-HT1C, 5-HT2C, 5-HTR2C, 5HTR2C, HTR1C

Probe Details

Chemical Probes Portal ID:

CPP1873

Probe alias:

NBOH-2C-CN

PubChem CID:

90489020

SMILES string:

COC1=CC(=C(C=C1CCNCC2=CC=CC=C2O)OC)C#N

InChi Key:

VWEDZTZAXHMZIL-UHFFFAOYSA-N

Physico-chemical properties

Solubility - preferred solvent:

Physiologic saline (0.9% NaCl)

Solubility - concentration:

3 mg/mL

Potential reactivities:

not known

NMR:

1H NMR (300 MHz, DMSO-d6) δ 10.31 (s, 1H), 9.25 (br s, 2H), 7.41 (dd, J = 7.5, 1.6 Hz, 1H), 7.20 (ddd, J = 8.2, 7.4, 1.6 Hz, 1H), 7.31 (s, 1H), 7.14 (s, 1H), 6.99 (dd, J = 8.2, 0.9 Hz, 1H), 6.81 (ddd, J = 7.5, 7.4, 0.9 Hz, 1H), 4.08 (s, 2H), 3.86 (s, 3H), 3.76 (s, 3H), 3.07 (s, 4H).

13C NMR (75 MHz, DMSO-d6) δ 155.9, 155.0, 150.7, 133.0, 131.5, 130.2, 118.9, 117.9, 116.3, 115.3, 114.7, 114.5, 98.3, 56.5, 56.3, 45.3, 44.9, 26.9.

Storage Recommendations

Storage (solution):

5°C

Storage sensitivities:

Stable for few weeks

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

0.81 nM (2A), 30 nM (2B), 42 nM (2C)

Potency assay:

Radioligand competition binding assay performed using: [3H]LSD (data shown), [3H]Cimbi-36, ([3H]ketanserin), ([3H]mesulergine)

PDB ID for probe-target interaction (3D structure):

6WHA

Off target activity

Potency assay (off target):

Broad profiling screens performed at Eurofins and PDSP

Probe Selectivity in Vitro:

25CN-NBOH also displayed appreciable binding affinity at 5-HT6 (Ki value of 310 nM), dopamine D4 (Ki value 2.9 μM), α2A, α2B, and α2C adrenergic (Ki values of 1.2–2.9 μM), H1 histamine (Ki value of 2.1 μM), κ opioid (Ki value of 4.2 μM), and sigma-1 and -2 (Ki values of 120–280 nM) receptors

In cell validation

On target activity in cells

Potency in cells:

EC50

2.4 nM HTR2A, 350 nM HTR2C

Potency assay (cells):

IP-One homogeneous time-resolved fluorescence assay

Off target activity in cells

Probe Selectivity in Cell:

Tested at a wide range of kinases and other enzymes in enzymatic assays, displaying no significant activity at 10 μM .
In addition 25CN-NBOH inhibited hERG with an IC50 value of 2.7 µM.

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

The cellular toxicity was investigated in a high-content screening assay, where the effects of 25CN-NBOH at concentrations up to 100 µM for six parameters: no effects were recorded.

In vivo validation

Organism:

male C57BL/6 mice

Dose:

0.1 to 10 mg/Kg

Route of delivery:

SubcutaneousIntraperitoneal

Organ of Interest

Organ (O):

Brain

Primary Reference

Reference (doi):

10.3389/fphar.2018.00017 10.1007/s00213-018-5118-y 10.1007/s00213-014-3739-3 10.1016/j.neuropharm.2016.03.038 10.1124/jpet.117.239905 10.1021/cn400216u

Reference (PMID):

29467649 30448990 25224567 27020041 28360333 24397362

Endorsed probe

The Chemical Probes Portal endorses this compound as chemical probe for use in cells and in model organisms.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

Need to be aware compound is active on multiple targets - HTR2A, HTR2B, and HTR2C - when drawing biological conclusions from studies carried out with this compound.

Important to measure drug concentrations in any efficacy studies to put other pharmacology into context: sigma-1 (Ki = 284 nM), α2C (Ki = 543 nM), 5-HT6 (Ki = 573 nM), sigma-2 (Ki = 575 nM), α2A (Ki = 803 nM)

Jul 25 2020 - 6:18pm

Mode of in vivo delivery i.p or s.c. @ 1.5 mg/kg is indicated by Buchborn 2018. Note potential for rapid desensitization in mouse models

Aug 13 2020 - 11:56am

25CN-NBOH

Probe Availability

Probe Information

Target Details

5-HT2A, HTR2, 5-HT1C, 5-HT2C, 5-HTR2C, 5HTR2C, HTR1C

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?

Search form

25CN-NBOH

Probe Availability

Probe Information

Target Details

5-HT2A, HTR2, 5-HT1C, 5-HT2C, 5-HTR2C, 5HTR2C, HTR1C

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?