Optimization of enzyme replacement therapy for treatment of homocystinuria
AssigneesThe Regents of the University of Colorado, a body corporate, Travere Therapeutics Switzerland GmbH
InventorsTomas Majtan, Jan P. Kraus, Erez M. Bublil, Frank Glavin, Marcia Sellos-Moura
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Abstract
The present invention provides a method of PEGylating a human truncated cystathionine β-synthase protein containing a mutation of a cysteine to a serine at amino acid position 15 (htCBS C15S). The htCBS C15S was PEGylated with one of 5 kDa, 10 kDa, or 20 kDa NHS ester PEG molecules. In-process monitoring of the PEGylation process was used in the method to reduce levels of unPEGylated htCBS C15S and htCBS C15S with insufficient PEGylation. Administration of the PEGylated htCBS C15S had efficacy throughout the course of treatment for homocystinuria.
Claims13
(canceled)
A method of PEGylating a human truncated cystathionine β-synthase (CBS) protein containing a mutation of a cysteine to a serine at amino acid position 15 of SEQ ID NO:
- 1 or a variant thereof (htCBS C15S), the method comprising:
- (a) conjugating the htCBS C15S with one or a plurality of NHS ester PEG molecules in solution at a molar excess of the NHS ester PEG molecules less than about 40-fold to create a batch, wherein the plurality of NHS ester PEG molecules are 5 kDa, 10 kDa, or 20 kDa NHS ester PEG molecules;
- (b) comparing a retention time from non-reduced capillary electrophoresis (NR-CE) analysis of the batch to a retention time from NR-CE analysis of htCBS C15S with acceptable PEGylation to identify insufficient PEGylation, wherein the batch with insufficient PEGylation has a retention time less than the retention time of the htCBS C15S with acceptable PEGylation;
- and
- (c) adding additional NHS ester PEG molecules to the batch to reduce the amount of insufficient PEGylation in the batch, thereby PEGylating the htCBS C15S.
-25. (canceled)
Dependencies are inferred from the claim text.