Intracellular antibody delivery

Patent metadata

Field	Value
Publication number	US-9732142-B2
Application number	US-99133009-A
Country	US
Kind code	B2
Filing date	May 14, 2009
Priority date	May 15, 2008
Publication date	Aug 15, 2017
Grant date	Aug 15, 2017

Abstract

The present invention concerns a composition comprising vesicles and encapsulated within the vesicles, an antibody, wherein the vesicles comprise an amphiphilic block copolymer having a hydrophilic and a hydrophobic block.
Methods of delivering the above compositions into cells are also described.

First claim

The invention claimed is:

1. A composition comprising nanovesicles and encapsulated within the aqueous core of the nanovesicles, an antibody, wherein the nanovesicles comprise an amphiphilic block copolymer having a hydrophilic block formed from 2-methacryloyloxy ethyl phosphorylcholine and a hydrophobic block formed from 2-(diisopropyl)amino ethyl methacrylate, the ratio of the degree of polymerization of the hydrophilic to hydrophobic block is in the range of 1:2.5 to 1:8, the nanovesicles have a diameter in the range of 50 to less than 1,000 nm, and the degree of polymerization of the hydrophilic block is about 20-25 and the degree of polymerization of the hydrophobic block is about 70-75.

2. A composition according to claim 1 , wherein one of the blocks comprises pendant groups which have a pK a in the range 3.0 to 6.9.

3. A composition according to claim 1 , wherein the antibody is capable of specific binding to an endogenous intracellular target.

4. A composition according to claim 1 , wherein the antibody is IgG antibody.

5. A composition according to claim 1 , wherein the antibody is IgG1 antibody.

6. A composition according to claim 1 , wherein the antibody is NFκB-p65 antibody.

7. A composition according to claim 1 , wherein the composition comprises a primary antibody and a secondary antibody.

8. A composition according to claim 7 , wherein the primary antibody is anti-Golgin-97(human) mouse IgG1 monoclonal CDF4 antibody.

9. A composition according to claim 7 , wherein the secondary antibody is goat Anti-human IgG antibody having a fluorescent label.

10. A composition according to claim 7 , wherein the composition comprises anti-Golgin-97(human) mouse IgG1 monoclonal CDF4 antibody as a primary antibody and goat Anti-human IgG antibody having a fluorescent label as a secondary antibody.

11. A composition according to claim 2 , wherein the hydrophobic block comprises pendent groups which have a pK a in the range of 3.0 to 6.9.

12. A composition according to claim 11 , wherein the pK a is in the range of 4.0 to 6.9.

13. A composition according to claim 2 , wherein the pK a is in the range of 4.0 to 6.9.

14. A method for forming a composition according to claim 1 , comprising the steps:
(i) dispersing the amphiphilic copolymer in an aqueous medium;
(ii) acidifying the composition formed in step (i);
(iii) adding the antibody to the acidified composition; and
(iv) raising the pH to around neutral to encapsulate the antibody.

15. A method according to claim 14 , comprising a preliminary step, before step (i), wherein the amphiphilic copolymer is dissolved in an organic solvent in a reaction vessel and the solvent is then evaporated to form a film on the inside of the reaction vessel.

16. An in vitro method of delivering an antibody into a cell comprising contacting a composition according to claim 1 with the cell.

17. A method according to claim 16 , wherein the cell is alive.

18. A method according to claim 16 , wherein the nanovesicles are taken up by the cell and once inside the cell, the nanovesicles dissociate and release antibody, which binds to an intracellular target.

19. A method of delivering an antibody to a patient in need thereof, comprising administering a composition according to claim 1 to the patient in need thereof.

20. A method according to claim 19 , wherein an antibody is delivered into a cell.

Assignees

• Lewis Andrew Lennard
• Battaglia Giuseppe
• Massignani Marzia
• Biocompatibles Uk Ltd

Inventors

• Lewis Andrew Lennard
• Battaglia Giuseppe
• Massignani Marzia

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• A61P31/12

  Antivirals · CPC title

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• A61P25/28

  for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia · CPC title

• C08F290/00

  Macromolecular compounds obtained by polymerising monomers on to polymers modified by introduction of aliphatic unsaturated end or side groups · CPC title