Small-molecule hydrophobic tagging of fusion proteins and induced degradation of same

Patent metadata

Field	Value
Publication number	US-9500653-B2
Application number	US-201113992076-A
Country	US
Kind code	B2
Filing date	Dec 6, 2011
Priority date	Dec 7, 2010
Publication date	Nov 22, 2016
Grant date	Nov 22, 2016

Abstract

The present invention includes compounds that are useful in perturbing or disrupting the function of a transmembrane or intracellular protein, whereby binding of a compound to the transmembrane or intracellular protein induces proteasomal degradation of the transmembrane or intracellular protein. The present invention further includes a method of inducing proteasomal degradation of a transmembrane or intracellular protein. The present invention further includes a method of identifying or validating a protein of interest as a therapeutic target for treatment of a disease state or condition.

First claim

The invention claimed is:

1. A compound of formula:
HYD-L R ,
wherein HYD is a hydrophobic group selected from the group consisting of HTL-13, HTL-34 and HTL-35:

wherein L R is a linker group having a reactive moiety selected from the group consisting of:

and
wherein the reactive moiety is capable of forming a covalent link between the HYD group and a target protein of interest.

2. The compound of claim 1 , wherein the target protein of interest comprises at least one selected from the group consisting of a haloalkane dehalogenase, O 6 -alkylguanine-DNA alkyltransferase, ACP synthase, SCP synthase, and SFP synthase.

3. The compound of claim 1 , wherein the protein of interest is a structural protein, receptor, enzyme, cell surface protein, a protein involved in catalytic activity, aromatase activity, motor activity, helicase activity, metabolic processes, antioxidant activity, proteolysis, biosynthesis, kinase activity, oxidoreductase activity, transferase activity, hydrolase activity, lyase activity, isomerase activity, ligase activity, enzyme regulator activity, signal transducer activity, structural molecule activity, binding activity, cell motility, membrane fusion, cell communication, regulation of biological processes, development, cell differentiation, response to stimulus, behavioral proteins, cell adhesion proteins, proteins involved in cell death, protein transporter activity, nuclear transport, ion transporter activity, channel transporter activity, carrier activity, permease activity, secretion activity, electron transporter activity, pathogenesis, chaperone regulator activity, nucleic acid binding activity, transcription regulator activity, extracellular organization and biogenesis activity or translation regulator activity.

4. A method of determining whether a protein of interest is a potential target of a bioactive agent or a drug target, the method comprising:
(a) providing a protein of interest covalently linked to a compound of claim 1 , wherein the hydrophobic moiety of the compound of claim 1 is capable of inducing degradation of the protein of interest intracellularly or on the surface of cells;
(b) exposing cells that utilize the protein of interest to the covalently labeled protein of step (a), wherein the labeled protein of step (a) is present within or on the surface of the cells;
(c) measuring the degradation of the covalently labeled protein in or on the surface of the cells; and
(d) determining if the degradation of the covalently labeled protein modulates the biological activity of the cells through a change in a phenotypic response of the cells consistent with the protein being a potential target for a bioactive agent or drug for a disease and/or condition modulated through the protein of interest.

5. A method of inducing degradation of a fusion protein protein of interest in a cell, the method comprising:
(a) expressing a protein of interest in a cell;
(b) reacting intracellularly or on the surface of the cell the expressed protein of interest with a compound comprising the compound of claim 1 , wherein the compound of claim 1 upon reaction with the protein of interest forms a covalent bond with the protein of interest to form a hydrophobically labeled protein; and
(c) allowing the hydrophobically labeled protein to degrade.

Assignees

• Crews Craig M
• Tae Hyun Seop
• Schneekloth Ashley R
• Neklesa Taavi
• Sundberg Thomas
• Univ Yale

Inventors

• Crews Craig M
• Tae Hyun Seop
• Schneekloth Ashley R
• Neklesa Taavi
• Sundberg Thomas

Classifications

• A61K47/52

  the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient · CPC title

• C07K2319/95

  containing a motif/fusion for degradation (ubiquitin fusions, PEST sequence) · CPC title

• C07K1/13

  Labelling of peptides · CPC title

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• G01N2500/10

  involving cells · CPC title