Granted Patent
B2
US 12,497,605 · App. 18/611,354 · Granted Dec 16, 2025
Engineered acid alpha-glucosidase variants
Inventors:
William Casey Hallows (San Francisco, CA); Rachel Cathleen Botham (Burlingame, CA); Yu Zhu (Newark, CA); Chinping Chng (Menlo Park, CA); Nikki Dellas (San Carlos, CA); Gjalt W. Huisman (Redwood City, CA); Moulay Hicham Alaoui Ismaili (San Mateo, CA); David William Homan (San Ramon, CA); Adam P. Silverman (San Carlos, CA); Jonathan Vroom (South San Francisco, CA); Jessica P. Lao (San Carlos, CA)
Assignee:
Crosswalk Therapeutics, Inc.
View Patent ↗
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Abstract
The present invention provides engineered acid alpha-glucosidase (GAA) polypeptides and compositions thereof. In some embodiments, the engineered GAA polypeptides have been optimized to provide increased expression, stability at neutral pH, and activity in cell lysates. The invention also provides methods for utilization of the compositions comprising the engineered GAA polypeptides for therapeutic and other purposes.
Claims (20)
1 . A recombinant acid alpha glucosidase comprising an amino acid sequence comprising at least 90% sequence identity to the sequence of residues 20 to 944 of SEQ ID NO: 3116, wherein the amino acid sequence has substitutions at positions L24W, L28S, L29T, P39Q, Q50V, A62L, P78E, D87E, L109D, S135Q, T150S, T266N, R267K, A437G, T486E, E522V, L569T, T692G, A711H, A750P, A812E, Q830K, L860F, L871E, R883H, Q894G, V913R, and S932A compared to SEQ ID NO: 2.
2 . The recombinant acid alpha glucosidase of claim 1 , wherein the recombinant acid alpha glucosidase is derived from a human acid alpha glucosidase.
3 . The recombinant acid alpha glucosidase of claim 1 , wherein the recombinant acid alpha glucosidase exhibits at least one improved property selected from: i) enhanced catalytic activity; ii) increased tolerance to pH 7; iii) increased tolerance to pH 4; iv) increased expression; v) increased uptake into cells; vi) increased enzymatic activity in cell lysates; vii) reduced immunogenicity; or a combination of any of i), ii), iii), iv), v), vi), or vii), as compared to SEQ ID NO: 2.
4 . A composition comprising at least one recombinant acid alpha glucosidase of claim 1 and a pharmaceutically acceptable carrier and/or excipient.
5 . A recombinant polynucleotide sequence encoding at least one recombinant acid alpha glucosidase of claim 1 .
6 . The recombinant polynucleotide sequence of claim 5 , wherein the polynucleotide sequence is selected from DNA, RNA, and mRNA and the polynucleotide sequence is codon-optimized.
7 . An expression vector comprising the recombinant polynucleotide sequence of claim 5 .
8 . The expression vector of claim 7 , wherein the recombinant polynucleotide sequence is operably linked to a control sequence that is a promoter.
9 . A host cell comprising the expression vector of claim 7 .
10 . The host cell of claim 9 , wherein the host cell is a mammalian cell.
11 . A method of producing a recombinant acid alpha glucosidase, comprising culturing the host cell of claim 9 , under conditions that the acid alpha glucosidase encoded by the recombinant polynucleotide is produced.
12 . A recombinant acid alpha glucosidase produced according to the method of claim 11 .
13 . A composition comprising the recombinant acid alpha glucosidase of claim 12 .
14 . A composition comprising at least one polynucleotide of claim 5 .
15 . A pharmaceutical composition, comprising the composition of claim 4 and a pharmaceutically acceptable carrier and/or excipient.
16 . A pharmaceutical composition comprising the recombinant polynucleotide of claim 5 and a pharmaceutically acceptable carrier and/or excipient.
17 . A method for treating symptoms of Pompe disease in a subject or ameliorating symptoms of Pompe disease in a subject, the method comprising the pharmaceutical composition of claim 16 .
18 . The method of claim 17 , wherein the symptoms of Pompe disease are ameliorated.
19 . The method of claim 17 , wherein the subject is an infant or child.
20 . The method of claim 17 , wherein the subject is an adult or young adult.
Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Aug 16, 2024
From: CODEXIS, INC.
To: CROSSWALK THERAPEUTICS, INC.
Reel/Frame 068660/0439 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Aug 2, 2024
From: HALLOWS, WILLIAM CASEY; BOTHAM, RACHEL CATHLEEN; ZHU, YU; CHNG, CHINPING; DELLAS, NIKKI; HUISMAN, GJALT W.; ALAOUI ISMAILI, MOULAY HICHAM; HOMAN, DAVID WILLIAM; SILVERMAN, ADAM P.; VROOM, JONATHAN; LAO, JESSICA P.
To: CODEXIS, INC.
Reel/Frame 068293/0746 →
Continuity (3)
Provisional Application
62951625
· Dec 20, 2019
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[cited by applicant]