Mannose-6-phosphate bearing peptides fused to lysosomal enzymes

Patent metadata

Field	Value
Publication number	US-10722559-B2
Application number	US-201515503223-A
Country	US
Kind code	B2
Filing date	Aug 11, 2015
Priority date	Aug 11, 2014
Publication date	Jul 28, 2020
Grant date	Jul 28, 2020

Abstract

A targeted therapeutic including a lysosomal enzyme and a lysosomal targeting moiety that is a peptide containing at least one N-linked glycosylation site. Methods of producing the targeted therapeutic may include nucleotide acids encoding the same and host cells co-expressing GNPT. Pharmaceutical compositions comprising the targeted therapeutic and methods of using the same to treat a lysosomal storage disease.

First claim

We claim:

1. A nucleic acid encoding a targeted therapeutic fusion protein comprising:
a lysosomal enzyme, and
a lysosomal targeting moiety,
wherein the lysosomal targeting moiety is a prosaposin peptide comprising an amino acid sequence of SEQ ID NO: 4, containing at least one N-linked glycosylation site, and
wherein the fusion protein, once administered in vivo, results in reduced lysosomal storage.

2. A vector comprising the nucleic acid sequence of claim 1 .

3. A host cell comprising the vector of claim 2 .

4. The host cell of claim 3 , wherein the host cell co-expresses N-acetylglucosamine-1-phosphoTransferase (GNPT).

5. The host cell of claim 4 , wherein the host cell comprises an exogenous nucleic acid encoding GNPT.

6. The host cell of claim 4 , wherein the host cell has activated expression of endogenous GNPT.

7. A method for producing a targeted therapeutic fusion protein, comprising:
a) culturing cells comprising a nucleic acid of claim 1 , wherein the cells co-express N-acetylglucosamine-1-phosphoTransferase (GNPT); and
b) recovering the fusion protein produced by the cells.

8. The nucleic acid of claim 1 , wherein the fusion protein comprises an amino acid sequence at least 90% identical to SEQ ID NO: 18.

9. The nucleic acid of claim 1 , wherein the fusion protein comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO. 23.

10. The nucleic acid of claim 1 , wherein the lysosomal targeting moiety and the lysosomal enzyme is fused via a linker.

11. The nucleic acid of claim 10 , wherein the linker comprises a sequence of

(SEQ ID NO.: 15)

GAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAG

GGGGGAP.

Assignees

• Shire Human Genetic Therapies

Inventors

• Concino Michael F
• Strack-Logue Bettina
• Meiyappan Muthuraman
• Norton Angela W
• Zhang Bohong
• Iskenderian Andrea
• Low Lieh Yoon
• Lundberg Dianna
• Romashko Alla
• Naimy Hicham

Classifications

• C07K14/4705

  stimulating, promoting or activating activity · CPC title

• C12N9/1288

  Transferases for other substituted phosphate groups (2.7.8) · CPC title

• A61K38/43 Primary

  Enzymes; Proenzymes; Derivatives thereof · CPC title

• A61K38/30

  Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2 {(insulin-like growth factor binding protein A61K38/1754)} · CPC title

• A61K38/47 Primary

  acting on glycosyl compounds (3.2), e.g. cellulases, lactases · CPC title