Soluble mTOR Complexes for High-Throughput Screening
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Solution Overview
Problem
Current methods for identifying modulators of mTORC1 and mTORC2 are hindered by difficulties in distinguishing between inhibited species, and there is a need for specific inhibitors to target these complexes effectively in treating proliferative and metabolic diseases.
Innovation Solution
Development of novel small molecule compounds that inhibit mTORC1 and mTORC2 in an ATP competitive manner, along with pharmaceutical compositions and methods for using these compounds to treat proliferative and metabolic diseases, including the use of purified mTORC1 and mTORC2 complexes for high-throughput screening.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If cell-based screens are used to discover mTOR inhibitors, then the screening can be performed using cellular systems, but the identification of inhibited species becomes ambiguous and difficult
Solution Approach 1:
The patent divides the mTOR complex into two distinct, functionally independent complexes: mTORC1 (containing Raptor) and mTORC2 (containing Rictor). Each complex can be independently purified, assayed, and inhibited by specific compounds. This segmentation allows unambiguous identification of inhibited species through selective inhibition of individual complexes, resolving the ambiguity present in cell-based screens.
Solution Approach 2:
The patent introduces purified recombinant protein complexes as intermediary assay systems between the inhibitor compounds and the cellular targets. These purified mTORC1 and mTORC2 complexes serve as mediators that can be directly assayed for inhibition without the complexity of whole cell systems, enabling precise measurement and unambiguous identification of inhibited species.
2Reliability
If rapamycin is used to inhibit mTORC1, then the complex is inhibited through FKBP12-rapamycin binding domain, but mTORC2 remains largely insensitive and distinguishing inhibited species becomes difficult
Solution Approach 1:
The patent segments mTOR inhibition into two independent pathways by separating mTORC1 and mTORC2 into distinct complexes with different protein compositions. mTORC1 contains Raptor while mTORC2 contains Rictor, allowing selective inhibition of each complex by different compounds. This segmentation enables reliable differentiation and measurement of inhibition for each species independently.
Solution Approach 2:
The patent applies local quality by developing inhibitors with specific binding affinities for different complexes. Compounds like rapamycin bind specifically to the FRB domain of mTORC1, while other compounds are designed to target mTORC2 specifically. This localized specificity allows distinct inhibition patterns that can be easily detected and measured for each complex.
3Reliability
If independent mTORC1 and mTORC2 complexes are targeted, then disease treatment specificity can be improved, but the complexity of developing and screening inhibitors increases
Solution Approach 1:
The patent segments the mTOR complex into two independently assayable units (mTORC1 and mTORC2) that can be purified and studied separately. This segmentation simplifies the screening process by allowing parallel assessment of compounds against each complex using standardized purified protein assays, reducing the overall complexity compared to cell-based approaches while maintaining therapeutic specificity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The compounds effectively inhibit mTORC1 and mTORC2, providing a therapeutic approach for treating diseases such as cancer and metabolic disorders by selectively targeting these complexes, enhancing treatment efficacy and specificity.
Implementation Method 1
The compounds may inhibit mTORC1 or mTORC2 in an ATP competitive manner
Data Source
AI summary
The present invention relates to small molecule modulators of mTORC1 and mTORC2, syntheses thereof, and intermediates thereto. Such small molecule modulators are useful in the treatment of proliferative diseases (e.g., benign neoplasms, cancers, inflammatory diseases, autoimmune diseases, diabetic retinopathy) and metabolic diseases. Novel small molecules are provided that inhibit one or more of mTORC1, mTORC2, and PI3K-related proteins. Novel methods of providing soluble mTORC1 and mTORC2 complexes are discussed, as well as methods of using the soluble complexes in a high-throughput manner to screen for inhibitory compounds.