Enhancement of intracellular delivery and tissue targeting of drugs and genes

Abstract

A method for enhancing intracellular delivery of effector molecules is provided. The method involves modifying selected antibodies with biotin and streptavidin, conjugating these antibodies with an effector molecule, and delivering the conjugated effector to an intracellular target specifically recognized by the antibody.

Description

INTRODUCTION [0001] This application is a continuation of U.S. application Ser. No. 10 / 446,422 filed May 28, 2003, which is a continuation of U.S. application Ser. No. 09 / 623,822 filed Oct. 25, 2000, which is the U.S. National Phase of PCT application Ser. No. PCT / US99 / 05279 filed Mar. 10, 1999, which claims the benefit of priority from U.S. Provisional Application Ser. No. 60 / 077,375 filed Mar. 10, 1998, each of which are herein incorporated by reference in their entirety.BACKGROUND OF THE INVENTION [0002] Targeting of drugs or genetic material to defined cells, tissues or organs increases the specificity and effectiveness of drug therapy and reduces the incidence of potentially harmful side effects. Intracellular delivery and proper intracellular processing are required for specific and effective therapeutic applications of certain classes of drugs including, but not limited to, immunotoxins, antioxidants, NO-donors, antibiotics, antisense oligonucleotides, nucleic acids and intra...

AI Technical Summary

Benefits of technology

[0012] An object of the present invention is to provide a method for enhancing cellular internalization of selected antibodies which comprises biotinylation of a selected antibody followed by conjugation of the biotinylated antibody with streptavidin. The modification of the antibody with streptavidin leads to an increase in the amount of antibody internalized.

[0013] Another object of the present invention is to provide a method for enhancing intracellular delivery of an effector to a selected target cell by conjugating the effector with a biotinylated, streptavidin-conjugated antibody that is targeted to the selected cell.

[0014] Another object of the present invention is to provide a method for enhancing accumulation of an antibody in a selected tissue by conjugating a biotinylated antibody with streptavidin. The modification of the antibody with streptavidin leads to an increase in the amount of antibody accumulated in the target tissue.

Problems solved by technology

Although these strategies may facilitate internalization, their applicability is restricted.

For example, none of these methods provides targeting of an effector to a specific cell, tissue, or organ, restricting the specificity and safety of the therapeutic agent.

Further, these methods utilize cellular mechanisms of internalization leading to accumulation of an effector in the lysosomes and ultimately resulting in degradation and inactivation of the effector compound.

Accumulation of these antibodies or antibody-conjugated effectors in lysosomes and subsequent lysosomal degradation restrict the applicability of the internalizable antibody as a carrier for intracellular delivery of drugs.

Therefore, these “poorly internalizable” antibodies are not useful for intracellular targeting.

However, these internalizable antibodies underwent massive intracellular degradation in lysosomes.

However, the total amount of anti-ACE binding sites in pulmonary endothelium was limited to 2×105 per cell (Muzykantov et al.

The limited number of binding sites, as well as significant intracellular degradation, limit the utility of this antibody system for intracellular targeting.

In pathological conditions associated with acute hypotonia, inhibition of ACE activity may lead to dangerous side effects, such as vascular collapse.