US-12622876-B2 - Methods of suppressing delivery of exosomes to liver and spleen

US12622876B2US 12622876 B2US12622876 B2US 12622876B2US-12622876-B2

Abstract

The instant application describes improved methods and compositions for the systemic delivery of therapeutic exosomes to a subject in need thereof. In certain embodiments, the current invention reduces the amount of exosomes delivered to liver, spleen and combinations thereof to allow greater distribution to other areas of the body such as, but not limited to, the brain, pancreas, lung, kidney, muscle. In certain embodiments, the methods involve the injection of one or multiple doses of non-therapeutic exosomes prior to the injection of a suitable therapeutic dose of exosomes with a therapeutic payload. Also included are methods to improve immune clearance of exosomes in subjects by inhibiting phagocytosis.

Inventors

Douglas E. Williams
John D. Kulman

Assignees

LONZA SALES AG

Dates

Publication Date: 20260512
Application Date: 20170821

Claims (12)

1 . A method of blocking the uptake of therapeutic exosomes in the liver and/or spleen of a subject, the method comprising: intravenously administering to the subject a first dose comprising non-therapeutic exosomes; and intravenously administering to the subject a second dose comprising therapeutic exosomes, wherein the intravenous administration of the first dose comprising the non-therapeutic exosomes causes reduced delivery of the therapeutic exosomes to an organ selected from the group consisting of the liver, spleen, and combinations thereof, and causes increased delivery of the therapeutic exosomes to an organ selected from the group consisting of the lung, small intestine, large intestine, stomach, pancreas, and combinations thereof, compared to delivery of the therapeutic exosomes intravenously administered at the same dose, but without prior intravenous administration of the first dose comprising non-therapeutic exosomes, wherein the second dose comprises an exosome quantity of at least about 10 10 exosomes/g by weight of the subject and the first dose is intravenously administered as a bolus dose comprising an exosome quantity that is at least 5 times greater than the second dose, wherein the therapeutic exosomes harbor at least one therapeutic payload or have been modified to have a desired therapeutic effect as compared to non-therapeutic exosomes, wherein the non-therapeutic exosomes do not harbor at least one therapeutic payload as compared to a therapeutic exosome, and wherein the therapeutic and non-therapeutic exosomes are derived from a HEK293 producer cell.
2 . The method according to claim 1 , wherein the therapeutic exosomes comprise a receiver.
3 . The method of claim 1 , wherein the second dose is intravenously administered at a period of time which is 15 minutes or greater after intravenous administration of the first dose.
4 . The method of claim 1 , wherein the non-therapeutic exosomes, the therapeutic exosomes or both the non-therapeutic and therapeutic exosomes comprise an imaging agent.
5 . The method of claim 1 , wherein the therapeutic exosomes comprise: (i) an RNA, (ii) a DNA, (iii) a polypeptide, (iv) a polysaccharide, (v) a lipid, (vi) a toxin, or (vii) any combination thereof.
6 . The method of claim 1 , wherein the therapeutic exosomes comprise a microRNA (miRNA), an siRNA, or an shRNA.
7 . The method of claim 1 , wherein the therapeutic exosomes comprise more than one distinct payload selected from the group consisting of: a peptide, a protein, a DNA, an siRNA, an miRNA, an shRNA, a polysaccharide, a lipid, a toxin, doxorubicin, daunorubicin, docetaxel, irinotecan, a taxane, a topoisomerase inhibitor, cyclophosphamide, a vinca alkaloid, cisplatin, a retinoid, a nucleotide analog, a kinase inhibitor, and a combination thereof.
8 . The method of claim 1 , wherein the exosomes are contacted with sialyltransferase prior to intravenous administration.
9 . A kit, comprising: (i) a first pharmaceutical composition comprising non-therapeutic exosomes; (ii) a second pharmaceutical composition comprising therapeutic exosomes and (iii) instructions for use according to the method of claim 1 .
10 . The method of claim 1 , wherein the therapeutic exosomes comprise doxorubicin, daunorubicin, docetaxel, irinotecan, a taxane, a topoisomerase inhibitor, cyclophosphamide, vinca alkaloid, cisplatin, a retinoid, a nucleotide analog, a kinase inhibitor, or any combination thereof.
11 . The method of claim 1 , wherein the second dose is intravenously administered as a continuous infusion.
12 . The method of claim 1 , wherein the second dose is intravenously administered in a plurality of administration steps.

Description

CROSS REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Patent Application No. 62/378,122 filed Aug. 22, 2016, which is hereby incorporated by reference in its entirety. BACKGROUND OF THE INVENTION Field of the Invention The instant application relates to methods and compositions for introducing exosomes to a subject in need thereof. In certain embodiments, the methods involve the injection of one or multiple doses of non-therapeutic exosomes prior to the injection of a suitable therapeutic dose of exosomes harboring a therapeutic payload. In certain embodiments, the methods are designed to partially block uptake of therapeutic exosomes in the liver and/or spleen so as to enable systemic delivery to other organs and tissues. Description of the Related Art Studies in mice have shown that the majority of injected exosomes in normal or diseased mice are deposited in the liver and spleen (Wiklander, O. et al. Extracellular vesicle in vivo biodistribution is determined by cell source, route of administration and targeting. Journal of Extracellular Vesicles. Apr. 20 2015). The preferential biodistribution of exosomes to these organs may lead to reduced delivery of exosomes to other tissues of interest or lead to toxic off-target side-effects. Therefore, there is a need for methods and compositions that reduce the amount of exosomes delivered to the liver and spleen to allow greater distribution of exosomes to targets located at other locations and tissue types in the body. SUMMARY OF THE INVENTION Disclosed herein are methods and compositions for the delivery of exosomes to a subject in need thereof. In certain aspects are methods of introducing exosomes to a subject, the method comprising, administering to the subject a first dose comprising non-therapeutic exosomes and administering to the subject a second dose comprising therapeutic exosomes. In an embodiment, the therapeutic exosomes optionally comprise a receiver. In certain embodiments, the non-therapeutic and therapeutic exosomes are optionally administered separately. In certain embodiments, the second dose is optionally administered at a period of time which is 15 minutes or greater after administration of the first dose. In certain embodiments, the second dose is administered at a period of time which is three hours or less after administration of the first dose. In certain aspects, the first dose optionally comprises an exosome quantity that is different from an exosome quantity of the second dose. In some aspects, the first dose optionally comprises an exosome quantity that is greater than an exosome quantity of the second dose. In certain embodiments, the first and second doses are optionally administered parenterally. In some embodiments, the administration is optionally intravenous administration. In certain embodiments, the first dose is optionally a bolus dose comprising an exosome quantity that is greater than an exosome quantity of the second dose, and wherein the second dose is administered as a continuous infusion. In certain embodiments, the first dose is optionally a bolus dose comprising an exosome quantity that is greater than an exosome quantity of the second dose, and the second dose is administered in a plurality of repeated administration steps. In certain embodiments, the first dose is administered in a plurality of repeated administration steps. In some embodiments, the second dose is administered in a time period ranging from 15 minutes to 3 hours after completion of the plurality of repeated administration steps. In certain embodiments, the non-therapeutic exosomes, the therapeutic exosomes or both the non-therapeutic and therapeutic exosomes optionally comprise an imaging agent. In some embodiments, the imaging agent is a fluorescent compound. In some embodiments, the imaging agent is a radioactive compound. In certain embodiments, the methods further comprise imaging the subject after administration of the non-therapeutic exosomes, the therapeutic exosomes or both the non-therapeutic and therapeutic exosomes and thereby detecting the location of the exosomes in the subject. In some embodiments, the imaging comprises magnetic resonance imaging. In certain embodiments, the therapeutic exosomes optionally comprise RNA. In certain embodiments, the therapeutic exosomes comprise microRNA (miRNA). In certain embodiments, the therapeutic exosomes comprise siRNA. In certain embodiments, the therapeutic exosomes optionally comprise DNA. In certain embodiments, the therapeutic exosomes optionally comprise a polypeptide. In certain embodiments, the therapeutic exosomes optionally comprise a small molecule. In certain embodiments, the therapeutic exosomes optionally comprise a large molecule biologic. In certain embodiments, the therapeutic exosomes optionally comprise more than one distinct payload. In certain embodiments, the payload comprises more than one type of payload selected from the group consisting of p