Granted Patent B2

US 12,644,132 · App. 17/779,510 · Granted Jun 2, 2026

Inventors: Sergei Zolotukhin (Gainesville, FL); Roland Wilfried Herzog (Gainesville, FL); Damien Marsic (Gainesville, FL); Moanaro Biswas (Fishers, IN)

Assignee: University of Florida Research Foundation, Incorporated

C12N15/86C12N15/1058C12N2750/14143

Abstract

Claims (24)

1 . A variant recombinant adeno-associated virus (rAAV) serotype 3B (AAV3B) capsid protein comprising each of the following sets of sequences and/or substitutions in the wild-type AAV3B VP1 sequence of SEQ ID NO: 1:

(a) STX 4 X 5 GTTGTX 8 X 9 LX 10 (SEQ ID NO: 7) in variable region (VR) IV wherein X 4 is P; X 5 is G; X 8 is N; X 9 is G; and X 10 is K;

(b) X 11 X 12 X 13 X 14 NNNSNFPWTAASX 15 in VR V, wherein X 11 is I or T; X 12 is A or P; X 13 is S; X 14 is D or Q; and X 15 is K or T;

(c) KDDX 16 X 17 X 18 in VR VI, wherein X 16 is E or D; X 17 is D; and X 18 is K or R; and

(d) one of QSSNTAPTTRTVND (SEQ ID NO: 6) or QNGRDNPTFRDVQH (SEQ ID NO: 8) in VR VIII.

2 . The variant of claim 1 comprising one or more of (a) STPGGTTGTNGLK (SEQ ID NO: 3) in VR IV, (b) IPSQNNNSNFPWTAASK (SEQ ID NO: 4) in VR V, (c) KDDDDR (SEQ ID NO: 9) in VR VI, and (d) QSSNTAPTTRTVND (SEQ ID NO: 6) in VR VIII.

3 . The variant of claim 1 further comprising GKQGAGRDNTEYDH (SEQ ID NO: 5) in VR VII.

4 . The variant of claim 1 further comprising the substitution Q263A.

5 . The variant of claim 1 further comprising one or more of the substitutions R460G, N494S, S551D, A553T, L555Y, and N557H in SEQ ID NO: 1.

6 . The variant of claim 1 further comprising the substitutions R460G, N494S, S551D, A553T, L555Y, and N557H in SEQ ID NO: 1.

7 . The variant of claim 1 , wherein the capsid protein comprises the amino acid sequence set forth in SEQ ID NO: 2.

8 . A variant recombinant AAV3B particle comprising the recombinant AAV capsid protein of claim 1 .

9 . The variant recombinant AAV3B particle of claim 8 , further comprising a nucleic acid comprising a transgene of interest.

10 . The variant recombinant AAV3B particle of claim 9 , wherein the transgene is selected from F8, ATP7B, GAA and UGT1A1.

11 . The variant recombinant AAV3B particle of claim 8 , wherein the nucleic acid is single stranded or self-complementary.

12 . A composition comprising a plurality of the variant recombinant AAV3B particle of claim 9 , optionally wherein the composition further comprises a pharmaceutical carrier, optionally wherein the plurality is in an amount of between 1×10 11 vgs/ml and 2×10 11 vgs/ml, or between 1×10 12 and 4×10 12 vgs/ml.

13 . A method of transducing a hepatic cell with a transgene of interest, the method comprising providing to the hepatic cell the variant recombinant AAV particle of claim 9 , wherein the hepatic cell is a human hepatocyte.

14 . A method of treating a disease or disorder comprising administering the variant recombinant AAV particle of claim 8 , to a subject in need thereof.

15 . The method of claim 14 , wherein the disease or disorder is selected from Wilson's Disease, hemophilia, a lysosomal storage disorder, and Crigler-Najjar syndrome.

16 . The method of claim 14 , wherein the step of administering provides an about 1.5-fold, a 2-fold, a 2.5-fold, a 3-fold, a 3.2-fold, a 3.5-fold, a 4-fold, a 5-fold, a 6-fold, a 10-fold, a 12-fold or a 15-fold decrease in seroreactivity to neutralizing anti-AAV antibodies in the subject, relative to a wild-type recombinant AAV3B particle, and/or wherein the step of administering provides about a 15%, a 30%, a 50%, a 100%, a 200%, a 300%, a 400%, a 500%, a 750%, or a 1000% increase in transduction of the transgene of interest in hepatic cells in the subject, relative to a wild-type recombinant AAV3B particle.

17 . The method of claim 14 , wherein the step of administering provides about a 15%, a 30%, a 50%, a 100%, or more than a 100% increase in transduction of the transgene of interest in hepatic cells in the subject, relative to a recombinant AAV3-ST particle.

18 . The method of claim 14 , wherein the subject is a primate, optionally wherein the subject is a human.

19 . The method of claim 14 further comprising re-administering the recombinant AAV particle.

20 . A method comprising administering the variant recombinant AAV particle of claim 8 , to a subject in need thereof whom has previously been administered the recombinant AAV particle.

Assignments (2)

Continuity (2)

References Cited (117)