Wortmannin derivatives as probes of cellular proteins and processes

A method of conducting a drug discovery business comprising:

• a. providing a wortmannin bait moiety being derivatized to a solid support or molecular or chemical tag for purifying or identifying the bait moiety;
• b. contacting the wortmannin bait moiety with a library of binding partners;
• c. identifying those members of the binding partner library which specifically bind to the wortmannin bait moiety;
• d. providing a reaction mixture including a binding partner identified in step (c) as able to specifically bind to the wortmannin bait moiety;
• e. contacting the binding partner with a test compound;
• f. determining if the test compound specifically binds to the binding partner;
• g. formulating a pharmaceutical preparation including one or more compounds identified in step (f) as able to inhibit or mimic the activity of a wortmannin moiety.

A method of conducting a drug discovery business comprising:

• a. identifying those members of a binding partner library which specifically bind to a wortmannin bait moiety;
• b. identifying compounds by their ability to agonize or antagonize a binding partner identified in step (a);
• c. conducting therapeutic profiling of a compound identified in step (b), or further analogs thereof, for efficacy and toxicity in animals;
• d. formulating a pharmaceutical preparation including one or more agents identified in step (iii) as having an acceptable therapeutic profile.

A method of conducting a target discovery business comprising:

• a. providing a wortmannin bait moiety being derivatized to a solid support or molecular or chemical tag for purifying or identifying the bait moiety;
• b. contacting the wortmannin bait moiety with a library of binding partners;
• c. identifying those members of the binding partner library which specifically bind to the wortmannin bait moiety;
• d. licensing, to a third party, the rights for drug development for a binding partner identified in step (c) as able to specifically bind to the wortmannin bait moiety.

A method of generating a pharmaceutical preparation including one or more compounds capable of binding to a binding partner binding to a wortmannin bait moiety, comprising:

• a. providing a wortmannin bait moiety being derivatized to a solid support or molecular or chemical tag for purifying or identifying the bait moiety;
• b. contacting the wortmannin bait moiety with a library of binding partners;
• c. identifying those members of the binding partner library which specifically bind to the wortmannin bait moiety;
• d. providing a reaction mixture including a binding partner identified in step (c) as able to specifically bind to the wortmannin bait moiety;
• e. contacting the binding partner with a test compound;
• f. determining if the test compound binds to the binding partner;
• g. formulating a pharmaceutical preparation including one or more compounds identified in step (f) as able to inhibit or mimic the activity of a wortmannin moiety.

A composition including a bait moiety represented in the general formula

• 6
• wherein:
• X, independently for each occurrence, represents O or S,
• R1 represents —(CH2)n—X—R5,
• R2, R4 and R5, independently, represent H or a C1-C6 alkyl,
• R3 represents S-L-,
• S represents a solid support or molecular or chemical tag for purifying or identifying the bait moiety,
• L represents a linker, and
• n is 0, 1, 2 or 3.

A method for profiling wortmannin-binding components comprising:

• a. providing a wortmannin bait moiety including wortmannin or an analog thereof being derivatized to a molecular or chemical tag for purifying or identifying the bait moiety;
• b. contacting the bait moiety with a library of cellular components, wherein the library of cellular components has been derived from cells that have been exposed to a first set of conditions, and identifying those members of the cellular component library which specifically bind to the bait moiety;
• c. contacting the bait moiety with a library of cellular components, wherein the library of cellular components has been derived from cells that have been exposed to a second set of conditions, and identifying those members of the cellular component library which specifically bind to the bait moiety;
• d. comparing the cellular components identified in (b), with the cellular components identified in (c), wherein a difference between the cell components identified in (b) and the cellular components identified in the (c) indicates that the difference in the conditions cause a change in the cellular component that can be used to profile the binding components.

A method for identifying the phosphorylation state of wortmannin binding partners comprising:

• a. providing a wortmannin bait moiety including wortmannin or an analog thereof being derivatized to a molecular or chemical tag for purifying or identifying the bait moiety;
• b. contacting the bait moiety with a library of cellular components that have been derived from cells that have been exposed to a first set of conditions, and identifying those members of the cellular component library which specifically bind to the bait moiety;
• c. contacting the bait moiety with a library of cellular components that have been derived from cells that have been exposed to a second set of conditions, and identifying those members of the cellular component library which specifically bind to the bait moiety;
• d. comparing the cellular components identified in (b), with the cellular components identified in (c), thereby identifying cellular components that differently phosphorylated between the first and second set of conditions.

A method for identifying the phosphorylation state of wortmannin binding partners comprising:

• a. providing a wortmannin bait moiety including wortmannin or an analog thereof being derivatized to a molecular or chemical tag for purifying or identifying the bait moiety;
• b. contacting the bait moiety with a library of cellular components that have been derived from differentiated cells, and identifying those members of the cellular component library which specifically bind to the bait moiety;
• c. contacting the bait moiety with a library of cellular components that have been derived from undifferentiated cells, and identifying those members of the cellular component library which specifically bind to the bait moiety;
• d. comparing the cellular components identified in (b), with the cellular components identified in (c), thereby identifying cellular components that are differently phosphorylated between differentiated and undifferentiated cells.

A method for identifying the phosphorylation state of wortmannin binding partners comprising:

• a. providing a wortmannin bait moiety including wortmannin or an analog thereof being derivatized to a molecular or chemical tag for purifying or identifying the bait moiety;
• b. contacting the bait moiety with a library of cellular components that have been derived from cancer cells, and identifying those members of the cellular component library which specifically bind to the bait moiety;
• c. contacting the bait moiety with a library of cellular components that have been derived from non-cancer cells, and identifying those members of the cellular component library which specifically bind to the bait moiety;
• d. comparing the cellular components identified in (b), with the cellular components identified in (c), thereby identifying cellular components that are differently phosphorylated between cancer and non-cancer cells.