Furin-cleavable delivery constructs
AssigneeThornhill Therapeutics, Inc.
InventorsThomas Carl Hunter, Randall J. Mrsny, Geoffrey Masuyer
Life of this patent
Estimating the term…
Abstract
The present disclosure provides furin-cleavable delivery constructs that include a transcytosing element that is derived from a mono-ADP-ribosyl transferase and a heterologous cargo that is coupled to the carrier. The carrier is capable of facilitating transport of the heterologous cargo across an epithelial cell via transcytosis. The heterologous cargo may be released from a remaining portion of the delivery construct upon cleavage by a furin protease. The constructs may be used to facilitate delivery of a cargo to a basolateral side of an epithelial membrane.
Claims21
-58. (canceled)
A furin-cleavable delivery construct for delivery of a heterologous cargo across a polarized epithelial cell via transcytosis, the delivery construct comprising:
- a transcytosing element derived from a domain I of a mono-ADP-ribosyl transferase (mART);
- a heterologous cargo coupled to the transcytosing element;
- and
- a furin-cleavable cleavage site, wherein the furin-cleavable cleavage site has a scissile bond that is positioned between the transcytosing element and the heterologous cargo such that cleavage of the scissile bond by a furin protease releases the heterologous cargo from the transcytosing element;
- wherein the scissile bond of the furin-cleavable cleavage site is positioned such that, upon cleavage of the scissile bond by the furin protease, a released N-terminal fragment comprising the transcytosing element lacks a C-terminal portion of a domain I, lacks a domain II, and lacks a domain III of a mART.
A method of delivering a cargo across an epithelial membrane, the method comprising:
- delivering a furin-cleavable delivery construct to an apical side of an epithelial membrane such that the cargo (1) transcytoses across an epithelial cell of the epithelial membrane and (2) is cleaved from the transcytosing element by a furin protease to deliver the cargo to a basolateral side of the epithelial membrane, wherein the furin-cleavable delivery construct comprises a transcytosing element derived from a domain I of a mono-ADP-ribosyl transferase (mART);
- a heterologous cargo coupled to the transcytosing element;
- and a furin-cleavable cleavage site, wherein the furin-cleavable cleavage site has a scissile bond that is positioned between the transcytosing element and the heterologous cargo such that cleavage of the scissile bond by a furin protease releases the heterologous cargo from the transcytosing element, and wherein the scissile bond of the furin-cleavable cleavage site is positioned such that, upon cleavage of the scissile bond by the furin protease, a released N-terminal fragment comprising the transcytosing element lacks a C-terminal portion of a domain I, lacks a domain II, and lacks a domain III of a mART.
A nucleic acid comprising (a) a polynucleotide sequence encoding a furin-cleavable delivery construct for delivery of a heterologous cargo across a polarized epithelial cell via transcytosis, the delivery construct comprising:
- a transcytosing element derived from a domain I of a mono-ADP-ribosyl transferase (mART);
- a heterologous cargo coupled to the transcytosing element;
- and
- a furin-cleavable cleavage site, wherein the furin-cleavable cleavage site has a scissile bond that is positioned between the transcytosing element and the heterologous cargo such that cleavage of the scissile bond by a furin protease releases the heterologous cargo from the transcytosing element;
- wherein the scissile bond of the furin-cleavable cleavage site is positioned such that, upon cleavage of the scissile bond by the furin protease, a released N-terminal fragment comprising the transcytosing element lacks a C-terminal portion of a domain I, lacks a domain II, and lacks a domain III of a mART;
- or
- (b) a reverse complement of said polynucleotide sequence of (a).
Dependencies are inferred from the claim text.