Givosiran is a subcutaneously administered, liver-targeted RNA interference (RNAi) therapeutic that has been approved for treating acute hepatic porphyria (AHP). Elevation in plasma homocysteine (hyperhomocysteinemia) has been reported in AHP patients, and treatment with givosiran has been reported to further increase homocysteine levels in some patients. The mechanism of homocysteine elevation during givosiran treatment is unknown, but has been hypothesized to be mediated by a reduction in activity of cystathionine β-synthase (CBS), which uses homocysteine as a substrate. A liquid chromatography-tandem mass spectrometry-based assay was adapted to measure circulating CBS activity. Using plasma collected from the Phase III ENVISION study, CBS activity was measured to directly evaluate whether it is associated with elevated homocysteine levels in givosiran-treated patients. CBS activity was reduced following givosiran treatment and both homocysteine and methionine levels were inversely correlated with CBS activity. Following administration of a supplement containing vitamin B6, a cofactor for CBS, in four patients during the trial, plasma CBS activity was found to increase, mirroring a corresponding decrease in homocysteine levels. These results support the hypothesis that elevated homocysteine levels following givosiran treatment result from a reduction of CBS activity and that vitamin B6 supplementation lowers homocysteine levels by increasing CBS activity.

Conclusions:

Using an LC–MS/MS-based assay,24 we found plasma CBS activity in AHP patients to be reduced following givosiran treatment in samples collected from the Phase III ENVISION clinical trial.6 Plasma homocysteine and methionine, previously reported to shift upward in givosiran-treated patients,11 were found to be inversely correlated with CBS activity. In four patients given supplements containing vitamin B6, CBS activity trended upward within 2–4 months following supplement initiation. Overall, these results support the hypothesis that elevated homocysteine levels following givosiran treatment result from reduction of CBS activity and can be mitigated by vitamin B6 supplementation.