All You Ever Wanted to Know About JWH-018

Is JWH-018 Legal?

Testing was performed by a qualified preclinical ADME Tox / ADMET contract testing lab.
Samples were analyzed by LC/MS/MS using either an Agilent 6410 mass spectrometer coupled with an Agilent 1200 HPLC and a CTC PAL chilled autosampler, all controlled by MassHunter software (Agilent), or an ABI2000 mass spectrometer coupled with an Agilent 1100 HPLC and a CTC PAL chilled autosampler, all controlled by Analyst software (ABI). After separation on a C18 reverse phase HPLC column (Agilent, Waters, or equivalent) using an acetonitrile-water gradient system.

This assays determines whether a test agent inhibits specific CYP450 enzymes.
If we use the same I/Ki ratio as the FDA prescribes for CYPs then the IC50 needs to be >30 µM for the potential for there to be an issue with hERG to exist, therefore JWH-018 is negative for hERG.
JWH-018 showed no cytotoxicity up to 250 然 in HepG2.
JWH-018 along with its metabolites was found to be negative in both GreenScreen HC and GreenScreen HC S9 (metabolism) genotoxic assays at concentrations up to 75 痢/ml.
No tissue abnormalities were observed at time of necropsy. There was no serious acute toxicities observed in the main organs.
JWH-018 distributes well throughout the rat. It is metabolized and eliminated normally and shows a half life of ~2 hours.

JWH-018 showed a bi-phasic distribution suggesting both distribution and elimination phases. The clearance was consistent of hepatic blood flow rates in a rat of (55 ml/min/kg). The volume of distribution for JWH-018 suggests that the drug is well distributed.

Fluorescent cytochrome P450 IC50 determination: Cytochrome P450 inhibition is measured using fluorogenic substrates. Test agents and nsubstrates are dissolved in acetonitrile for this assay, as DMSO significantly inhibits some cytochrome P450s. Assays were performed at 37 °C using commercially available recombinant human cytochrome P450 expressed in insect cells.

Guidance Document on Using In Vitro Data to Estimate In Vivo Starting Doses for Acute Toxicity. Based on Recommendations from an International Workshop Organized by the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) and the National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) National Institute of Environmental Health Sciences National Institutes of Health U.S. Public Health Service Department of Health and Human Services.

GreenScreen HC, with S9 Genotoxicity Assay: A dilution series of each test compound is generated in a 96-well microplate. Two strains of cultured human lymphoblastoid TK6 cells are used: the test strain (GenM-T01) and the non-fluorescent control strain (GenM-C01).

Test substance concentrations for in vitro studies should span a broad range, covering and exceeding the anticipated maximal therapeutic plasma concentration. Ascending concentrations should be tested until a concentration-response curve has been characterized or physicochemical effects become concentration-limiting.