	US 12,077,773 B2
	Compositions and methods for treating non-age-associated hearing impairment in a human subject
Emmanuel John Simons, Boston, MA (US); and Robert Ng, Boston, MA (US)
Assigned to Akouos, Inc., Boston, MA (US)
Filed by Akouos, Inc., Boston, MA (US)
Filed on Oct. 11, 2023, as Appl. No. 18/485,178.
Application 18/485,178 is a continuation of application No. 17/816,305, filed on Jul. 29, 2022, granted, now 11,807,867.
Application 17/816,305 is a continuation of application No. PCT/US2021/018919, filed on Feb. 19, 2021.
Claims priority of provisional application 62/979,792, filed on Feb. 21, 2020.
Prior Publication US 2024/0093237 A1, Mar. 21, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/86 (2006.01); A61K 48/00 (2006.01); A61P 27/16 (2006.01); C07K 14/47 (2006.01)

CPC C12N 15/86 (2013.01) [A61P 27/16 (2018.01); C07K 14/47 (2013.01); A61K 48/00 (2013.01); C12N 2750/14143 (2013.01); C12N 2750/14144 (2013.01)]

25 Claims

1. A plurality of recombinant adeno-associated (rAAV) vectors comprising:

a) a first rAAV vector comprising, in order of 5′ to 3′:

(i) a 5′ ITR sequence of SEQ ID NO: 97;

(ii) a CAG promoter;

(iii) a 5′ OTOF coding region comprising exons 1 to (and through) 21 of OTOF cDNA, wherein the 5′OTOF coding region is at least 80% identical to SEQ ID NO: 101 and encodes the same amino acid sequence as encoded by SEQ ID NO: 101;

(iv) a SD intron sequence of SEQ ID NO: 102;

(v) an AK recombinogenic sequence of SEQ ID NO: 103; and

(vi) a 3′ ITR sequence of SEQ ID NO: 104; and

b) a second rAAV vector comprising, in order of 5′ to 3′:

(i) a 5′ ITR sequence of SEQ ID NO: 97;

(ii) an AK recombinogenic sequence of SEQ ID NO: 103;

(iii) a SA intron sequence of SEQ ID NO: 106;

(iv) a 3′ OTOF coding region that comprises exons 22 to (and through) exon 48 of OTOF cDNA, wherein the 3′OTOF coding region is at least 80% identical to SEQ ID NO: 107 and encodes the same amino acid sequence as encoded by SEQ ID NO: 107;

(v) a polyA sequence; and

(vi) a 3′ ITR sequence of SEQ ID NO: 104.

17. A composition comprising a first rAAV vector encapsidated by an Anc80 capsid and a second rAAV vector encapsidated by an Anc80 capsid, wherein:

a) the first rAAV vector comprises, in order of 5′ to 3′:

(i) a 5′ ITR sequence of SEQ ID NO: 97,

(ii) a CAG promoter,

(iii) a 5′ OTOF coding region that comprises exons 1 to (and through) 21 of OTOF cDNA, wherein the 5′ OTOF coding region is at least 80% identical to SEQ ID NO: 101 and encodes the same amino acid sequence as encoded by SEQ ID NO: 101,

(iv) a SD intron sequence of SEQ ID NO: 102,

(v) an AK recombinogenic sequence of SEQ ID NO: 103, and

(vi) a 3′ ITR sequence of SEQ ID NO: 104; and

b) the second rAAV vector comprises, in order of 5′ to 3′:

(i) a 5′ ITR sequence of SEQ ID NO: 97,

(ii) an AK recombinogenic sequence of SEQ ID NO: 103,

(iii) a SA intron sequence of SEQ ID NO: 106,

(iv) a 3′ OTOF coding region that comprises exons 22 to (and through) exon 48 of OTOF cDNA, wherein the 3′ OTOF coding region is at least 80% identical to SEQ ID NO: 107 and encodes the same amino acid sequence as encoded by SEQ ID NO: 107,

(v) a polyA sequence, and

(vi) a 3′ ITR sequence of SEQ ID NO: 104;

wherein the composition further comprises one or more pharmaceutically acceptable carriers, diluents, or excipients.