US 12,281,323 B2
Treatment for restoring ureagenesis in carbamoyl phosphate synthetase 1 deficiency
Gerald Lipshutz, Los Angeles, CA (US); and Matthew Nitzahn, Los Angeles, CA (US)
Assigned to THE REGENTS OF THE UNIVERSITY OF CALIFORNIA, Oakland, CA (US)
Appl. No. 17/601,557
Filed by The Regents of the University of California, Oakland, CA (US)
PCT Filed Apr. 15, 2020, PCT No. PCT/US2020/028303
§ 371(c)(1), (2) Date Oct. 5, 2021,
PCT Pub. No. WO2020/214694, PCT Pub. Date Oct. 22, 2020.
Claims priority of provisional application 62/833,853, filed on Apr. 15, 2019.
Prior Publication US 2022/0162639 A1, May 26, 2022
Int. Cl. C12N 15/86 (2006.01); C12N 9/00 (2006.01)
CPC C12N 15/86 (2013.01) [C12N 9/93 (2013.01); C12N 2750/14143 (2013.01); C12N 2750/14144 (2013.01); C12N 2800/22 (2013.01); C12Y 603/04016 (2013.01)] 11 Claims
 
1. A method of making a pharmaceutical composition comprising combining together in an aqueous formulation:
at least one adeno-associated viral vector comprising a carbamoyl phosphate synthetase 1 polynucleotide sequence (SEQ ID NO: 2); and
a pharmaceutical excipient selected from the group consisting of:
a preservative, a tonicity adjusting agent, a detergent, a viscosity adjusting agent, a sugar or a pH adjusting agent, wherein:
the vector and the carbamoyl phosphate synthetase 1 polynucleotide sequence are selected such that when the adeno-associated viral vector infects a human liver cell, carbamoyl phosphate synthetase 1 protein is expressed;
the method combines two adeno-associated viral vectors comprising:
a first adeno-associated viral vector comprising a first segment of a codon optimized carbamoyl phosphate synthetase 1 polynucleotide (SEQ ID NO: 2); and
a second adeno-associated viral vector comprising a second segment of a codon optimized carbamoyl phosphate synthetase 1 polynucleotide (SEQ ID NO: 2); wherein:
the first adeno-associated viral vector comprising the first segment of a codon optimized carbamoyl phosphate synthetase 1 polynucleotide and the second adeno-associated viral vector comprising the second segment of a codon optimized carbamoyl phosphate synthetase 1 polynucleotide are selected so that:
the first and second segments of the carbamoyl phosphate synthetase 1 polynucleotides overlap such that, following first and second adeno-associated viral vector infection of the human liver cell, the carbamoyl phosphate synthetase 1 polynucleotides concatemerize via homologous recombination so as to reconstitute a carbamoyl phosphate synthetase 1 gene that expresses the carbamoyl phosphate synthetase 1 protein in the liver cell; and
the first segment of the carbamoyl phosphate synthetase 1 polynucleotide is selected to comprise the sequence ATGCCTCAGA.