US 12,031,977 B2
Ex vivo system for determining multiple drug-drug transporter interactions and methods of use thereof
Robert S. Langer, Newton, MA (US); Carlo Giovanni Traverso, Newton, MA (US); Yunhua Shi, Belmont, MA (US); Vance Soares, Boston, MA (US); and Daniel Reker, Durham, NC (US)
Assigned to Massachusetts Institute of Technology, Cambridge, MA (US); and The Brigham and Women's Hospital, Inc., Boston, MA (US)
Filed by Massachusetts Institute of Technology, Cambridge, MA (US); and The Brigham and Women's Hospital, Inc., Boston, MA (US)
Filed on Dec. 10, 2020, as Appl. No. 17/118,267.
Claims priority of provisional application 62/946,336, filed on Dec. 10, 2019.
Prior Publication US 2021/0231645 A1, Jul. 29, 2021
Int. Cl. G01N 33/50 (2006.01)
CPC G01N 33/5088 (2013.01) [G01N 33/5044 (2013.01)] 10 Claims
 
1. An ex vivo system for use in determining multiplex interactions between a candidate drug and two or more drug transporters in an intestinal tissue explant, the system comprising:
(i) an intestinal tissue explant in planar contact with a substrate, wherein the intestinal tissue explant comprises intestinal epithelium from a large, non-human, mammalian gastrointestinal tract, wherein the intestinal epithelium comprises epithelial cells having a polarity in the tissue explant and two or more drug transporters, and wherein the tissue explant provides a luminal surface and a basolateral surface; and
(ii) two or more agents for use in reducing or eliminating expression of the two or more drug transporters in the intestinal tissue explant, wherein the two or more drug transporters are on the luminal surface of the tissue explant and selected from: p-glycoprotein (p-gp), breast cancer resistance protein (BCRP), multidrug resistance 2 (MRP2), monocarboxylate transporter 1 (MCT1), and peptide transporter 1 (PEPT1), wherein the two or more agents are two or more siRNA molecules selected from the group consisting of:
(1) a p-gp targeting siRNA molecule comprising a sense strand comprising the nucleotide sequence set forth in SEQ ID NO: 91 and an antisense strand comprising the nucleotide sequence set forth in SEQ ID NO: 92;
(2) a MRP2 targeting siRNA molecule comprising a sense strand comprising the nucleotide sequence set forth in SEQ ID NO: 93 and an antisense strand comprising the nucleotide sequence set forth in SEQ ID NO: 94;
(3) a BCRP targeting siRNA molecule comprising a sense strand comprising the nucleotide sequence set forth in SEQ ID NO: 95 and an antisense strand comprising the nucleotide sequence set forth in SEQ ID NO: 96;
(4) a PEPT1 targeting siRNA molecule comprising a sense strand comprising the nucleotide sequence set forth in SEQ ID NO: 99 and an antisense strand comprising the nucleotide sequence set forth in SEQ ID NO: 100; and
(5) a MCT1 targeting siRNA molecule comprising a sense strand comprising the nucleotide sequence set forth in SEQ ID NO: 101 and an antisense strand comprising the nucleotide sequence set forth in SEQ ID NO: 102,
wherein the system provides for multiplex interactions between the candidate drug and the two or more drug transporters to be determined by contacting the intestinal tissue explant with the candidate drug before and after expression of the two or more drug transporters is reduced or eliminated in the intestinal tissue explant.