Dolutegravir

Biological Activity

Dolutegravir (GSK1349572) is a novel potent integrase inhibitor with an IC50 of 2.7 nM in vitro. Dolutegravir (GSK1349572) is a compound for the treatment of HIV infection. Dolutegravir (GSK1349572) exhibited in vitro activity against most clinical isolates obtained from patients failing RAL-based therapy. In PBMC cell assay, Dolutegravir (GSK1349572) inhibited HIV integrase with the antiviral IC50=0.51 and IC90=2.0 nM. In MT-4 antiviral assays, potency shift extrapolated to 100% human serum was 75-fold providing PA-IC90=152 nM in PBMCs. Dolutegravir (GSK1349572) had a markedly different resistance profile as evidenced by limited cross-resistance to RAL-resistant molecular clones, by its in vitro activity against clinical isolates obtained from patients failing RAL-based therapy, and in selecting different mutations with low level FC during serial passage. In vitro experiments support the potential for Dolutegravir (GSK1349572) to have a higher genetic barrier to resistance when compared to 1st generation integrase inhibitors.

Protocol (for reference only)

Cell Experiment
Cell lines	MT-4
Preparation method	Growing MT-4 cells exponentially at a density of 500000 or 600000 /mL are infected with HIV-1 strain IIIB at a viral multiplicity of infection of 0.001 or a 50% tissue culture infective dose of 4 to 10. Then aliquoting the cells to 96-well plates in the presence of varying concentrations of S/GSK1349572. After incubation for 4 or 5 days, using the yellow tetrazolium MTT reagent [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide] to determine antiviral activity by a cell viability assay that either measured bioluminescence with a CellTiter-Glo luminescent reagent or measured absorbance at 560 and 690 nm .
Concentrations	0 to 10 μM
Incubation time	4 days or 5 days

Animal Experiment
Animal models
Formulation
Dosages
Administration

Chemical Information

Molecular Weight	419.38
Formula	C20H19F2N3O5
CAS Number	1051375-16-6
Solubility (25°C)	DMSO 60 mg/mL
Storage	Powder          -20°C   3 years ;  4°C   2 years In solvent       -80°C   6 months ;  -20°C   1 month

Conversion of different model animals based on BSA (PMID: 27057123)

Species	Mouse	Rat	Rabbit	Guinea pig	Hamster	Dog
Weight (kg)	0.02	0.15	1.8	0.4	0.08	10
Body Surface Area (m2)	0.007	0.025	0.15	0.05	0.02	0.5
Km factor	3	6	12	8	5	20

Animal A (mg/kg) = Animal B (mg/kg) multiplied by	Animal B Km
	Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the K_m factor for a mouse and then divide by the K_m factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Sharon L Walmsley, et al. N Engl J Med. Dolutegravir plus abacavir-lamivudine for the treatment of HIV-1 infection

[2] Parul Patel, et al. J Antimicrob Chemother. Pharmacokinetics of the HIV integrase inhibitor S/GSK1349572 co-administered with acid-reducing agents and multivitamins in healthy volunteers

[3] Ivy Song, et al. Br J Clin Pharmacol. Effect of atazanavir and atazanavir/ritonavir on the pharmacokinetics of the next-generation HIV integrase inhibitor, S/GSK1349572

[4] Charlotte Charpentier, et al. AIDS. In-vitro phenotypic susceptibility of HIV-2 clinical isolates to the integrase inhibitor S/GSK1349572