TECHNICAL FIELD

The present invention relates to the field of biological condensates.

BACKGROUND

In addition to membrane-bound organelles, such as mitochondria, lysosomes, and the endoplasmic reticulum, cells contain distinct sub-compartments that do not comprise a membrane between them and their immediate surrounding solution. Numerous of these membrane-less molecular assemblies have been shown to be formed through a process termed liquid-liquid phase separation or condensation. During this process, a solution comprising biological macromolecules separates into different phases, a condensate that is enriched in some of those macromolecules and a surrounding phase that is relatively depleted in those macromolecules. A number of cellular condensates have been recognized. In addition, phase-separated condensates can be formed outside of the cell, such as in solution or extracellularly (Alberti et al.,J Mol Biol,430(23), 2018, 4806-4820; Muiznieks et al.,J Mol Biol,430(23), 2018, 4741-4753). However, little or nothing is known about the mechanisms governing the partitioning of compounds into or the exclusion of compounds from condensates or the differences in the partitioning of compounds among various condensates.

Various condensates are known to be important for modulating cellular processes. For example, a condensate can bring together molecules at an elevated concentration to accelerate reactions inside the condensate or can sequester molecules in the condensate, reducing their concentration in the surrounding medium. Aberrant condensate function has also been implicated in various human diseases, such as neurodegenerative and proliferative diseases (Naumann et al.,Nat Commun,9(1), 2018, 335; Wegmann et al.,EMBO J,37(7), 2018, e98049; Aguzzi et al.,Trends Cell Biol,26(7), 2016, 547-558). However, in addition to the lack of understanding of the mechanisms governing condensate partitioning of compounds, there is little or nothing known regarding how to identify test compounds that interact with condensates, how to design compounds to improve interactions with condensates, and how such information can be used to improve treatment of diseases.

All references cited herein, including patent applications and publications, are incorporated by reference in their entirety.

BRIEF SUMMARY

In some aspects, provided herein are methods of determining a partition characteristic of a test compound in a target condensate, the method comprising: (a) combining the test compound and a composition comprising the target condensate and an extra-condensate solution; (b) determining the amount of the test compound in the target condensate, thereby determining the partition characteristic of the test compound in the target condensate. In some embodiments, the methods further comprise causing the formation of the target condensate prior to step (a).

In some aspects, provided herein are methods of determining a partition characteristic of a test compound in a target condensate, the method comprising: (a) adding the test compound to a composition comprising a target condensate and an extra-condensate solution; and (b) determining the amount of the test compound in the target condensate, thereby determining the partition characteristic of the test compound in the target condensate. In some embodiments, the methods further comprise causing the formation of the target condensate prior to step (a).

In some aspects, provided herein are methods of determining a partition characteristic of a test compound in a target condensate, the method comprising: (a) causing the formation of the target condensate in the presence of the test compound to obtain a composition comprising the target condensate and an extra-condensate solution; and (b) determining the amount of the test compound in the target condensate, thereby determining the partition characteristic of the test compound in the target condensate.

In some aspects, provided herein are methods of determining a partition characteristic of a test compound in a target condensate, the method comprising: (a) adding the test compound to a composition comprising precursor molecules; (b) causing the formation of the target condensate to obtain a composition comprising the target condensate and an extra-condensate solution; and (c) determining the amount of the test compound in the target condensate, thereby determining the partition characteristic of the test compound in the target condensate. In some embodiments, the methods further comprise combining the test compound and a precursor composition comprising precursor molecules prior to step (a). In some embodiments, the methods further comprise adding the test compound to a precursor composition comprising precursor molecules prior to step (a).

In some embodiments, the methods further comprise determining the amount of the test compound in the extra-condensate solution. In some embodiments, the amount of the test compound in the target condensate is determined prior to, simultaneously with, or after the amount of the test compound in the extra-condensate solution is determined. In some embodiments, the methods further comprise determining the ratio of the amount of test compound in the target condensate and the amount of test compound in the extra-condensate solution. In some embodiments, the methods further comprise separating the target condensate from the extra-condensate solution. In some embodiments, the methods further comprise identifying the target condensate prior to determining the amount of test compound in the target condensate.

In some embodiments, dysregulation of the target condensate is associated with a disease. In some embodiments, the methods further comprise characterizing the target condensate by identifying one or more macromolecules comprised therein. In some embodiments, the identifying comprises determining the amount of the one or more macromolecules in the target condensate. In some embodiments, the methods further comprise determining the ratio of the amount of test compound in the target condensate and the amount of the one or more macromolecules in the target condensate. In some embodiments, the target condensate comprises a protein comprising an intrinsically disordered sequence. In some embodiments, the methods further comprise labeling the target condensate in order to visualize the target condensate. In some embodiments, the target condensate is labeled with a radioactive label, a colorimetric label, a chemically-reactive label, or a fluorescent label.

In some embodiments, the composition comprises a cell. In some embodiments, the cell is a microorganism or an animal cell. In some embodiments, the cell comprises a condensate that is determined to be dysregulated. In some embodiments, the cell has one or more features of a neurodegenerative or proliferative disease.

In some embodiments, the target condensate is a cellular condensate. In some embodiments, cellular condensate is a cleavage body, a P-granule, a histone locus body, a multivesicular body, a neuronal RNA granule, a nuclear gem, a nuclear pore, a nuclear speckle, a nuclear stress body, a nucleolus, a Oct1/PTF/transcription (OPT) domain, a paraspeckle, a perinucleolar compartment, a PML nuclear body, a PML oncogenic domain, a polycomb body, a processing body, a signaling cluster, a Sam68 nuclear body, a stress granule, or a splicing speckle. In some embodiments, the target condensate is in a cell. In some embodiments, the cell is a microorganism or an animal cell. In some embodiments, the cell has one or more features of a neurodegenerative or proliferative disease. In some embodiments, the extra-condensate solution is intracellular fluid. In some embodiments, the intracellular fluid is cytosol or nucleosol.

In some embodiments, the target condensate is not in a cell. In some embodiments, the target condensate is an extracellular condensate. In some embodiments, the extra-condensate solution is extracellular fluid. In some embodiments, the extracellular fluid is interstitial fluid.

In some embodiments, the method is a cell free assay method. In some embodiments, the composition does not comprise a cell. In some embodiments, the composition comprises one or more of: a macromolecule, a salt, and a buffer.

In some embodiments, the composition comprises two or more target condensates. In some embodiments, the methods further comprise repeating the steps of the method for one or more additional condensates.

In some embodiments, the test compound is small molecule, a polypeptide, or a nucleic acid. In some embodiments, the test compound comprises a test compound label. In some embodiments, the test compound label is a radioactive label, a colorimetric label, a chemically-reactive label, or a fluorescent label. In some embodiments, the test compound label is a fluorescent label. In some embodiments, the amount of the test compound is determined by detecting the test compound label. In some embodiments, the amount of the test compound is determined by mass spectrometry, liquid chromatography, quantitative fluorescent microscopy and spectroscopy, nuclear magnetic resonance spectroscopy, Raman spectroscopy, and/or ultraviolet-visible spectrophotometry.

In some aspects, provided herein are methods of determining the partition characteristics of a plurality of test compounds in a target condensate, the method comprising performing a method of determining the partition characteristic described herein with a plurality of test compounds. In some embodiments, the methods further comprise comparing the partition characteristics of a subset or all of the plurality of test compounds in the target condensate. In some embodiments, the methods further comprise identifying test compounds that have the same or similar partition characteristics in a target condensate. In some embodiments, the methods further comprise identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar partition characteristics. In some embodiments, the methods further comprise determining the partition characteristic in a target condensate for one or more additional test compounds that comprise the identified characteristic. In some embodiments, the methods further comprise determining the partition characteristic in a target condensate for one or more additional test compounds that do not comprise the identified characteristic.

In some aspects, provided herein are methods of determining a relative partition characteristic of a test compound in a target condensate, the method comprising: (i) determining the partition characteristic of the test compound by performing a method of determining a partition characteristic described herein with the test compound; (ii) determining the partition characteristic of a reference compound by performing a method of determining a partition characteristic described herein with the reference compound; and (iii) calculating the ratio of the partition characteristics determined in (i) and (ii), thereby determining the relative partition characteristic of the test compound in the target condensate. In some embodiments, the test compound comprises a test compound label. In some embodiments, the reference compound is the test compound label.

In some aspects, provided herein are methods of determining relative partition characteristics of a plurality of test compounds in a target condensate, the method comprising: (1) performing a method of determining a relative partition characteristic of a test compound in a target condensate; and (2) repeating steps (i) and (iii) with a plurality of test compounds. In some embodiments, the methods further comprise comparing the relative partition characteristics in the target condensate of a subset or all of the plurality of test compounds. In some embodiments, the methods further comprise identifying test compounds that have the same or similar relative partition characteristics in the target condensate. In some embodiments, the methods further comprise identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar relative partition characteristics. In some embodiments, the methods further comprise determining the relative partition characteristic in the target condensate for one or more additional test compounds that comprise the identified characteristic. In some embodiments, the methods further comprise determining the relative partition characteristic in the target condensate for one or more additional test compounds that do not comprise the identified characteristic.

In some aspects, provided herein are methods of determining a condensate preference profile of a test compound, the method comprising: (a) determining the partition characteristic of the test compound in a first target condensate according to a method disclosed herein; (b) determining the partition characteristic of the test compound in a second target condensate according to a method disclosed herein; and (c) calculating a ratio of the partition characteristic of the test compound determined in the first target condensate and the second target condensate, thereby determining the condensate preference profile of the test compound. In some embodiments, the first target condensate and the second target condensate are in the same composition. In some embodiments, the first target condensate and the second target condensate are in different compositions. In some embodiments, the partition characteristic of the test compound in the first target condensate is determined prior to, simultaneously with, or after the partition characteristic of the test compound in the second target condensate is determined.

In some aspects, provided herein are methods of determining a condensate preference profile of a test compound, the method comprising: (a) determining the relative partition characteristic of the test compound in a first target condensate according to a method disclosed herein; (b) determining the relative partition characteristic of the test compound in a second target condensate according to a method disclosed herein; and (c) calculating a ratio of the partition characteristic of the test compound determined in the first target condensate and the second target condensate, thereby determining the condensate preference profile of the test compound. In some embodiments, the first target condensate and the second target condensate are in the same composition. In some embodiments, the first target condensate and the second target condensate are in different compositions. In some embodiments, the relative partition characteristic of the test compound in the first target condensate is determined prior to, simultaneously with, or after the relative partition characteristic of the test compound in the second target condensate is determined.

In some aspects, provided herein are methods of determining a condensate preference profile of a test compound, the method comprising: (a) adding the test compound to a composition comprising a first target condensate and a second target condensate; (b) determining the amount of the test compound in the first target condensate; (c) determining the amount of the test compound in the second target condensate; and (d) calculating a ratio of the amount of the test compound determined in the first target condensate and the second target condensate, thereby determining the condensate preference profile of the test compound. In some embodiments, the methods further comprise causing the formation of the first target condensate and/or the second target condensate prior to step (a).

In some aspects, provided herein are methods of determining a condensate preference profile of a test compound, the method comprising: (a) adding the test compound to a composition comprising precursor molecules; (b) causing the formation of a first target condensate and a second target condensate in the composition; (c) determining the amount of the test compound in the first target condensate; (d) determining the amount of the test compound in the second target condensate; and (e) calculating a ratio of the amount the test compounds determined in the first target condensate and the second target condensate, thereby determining the condensate preference profile of the test compound. In some embodiments, the amount of the test compound in the first target condensate is determined prior to, simultaneously with, or after the amount of the test compound in the second target condensate is determined.

In some embodiments, the methods further comprise separating the first target condensate and the second target condensate from the composition. In some embodiments, the methods further comprise identifying the first target condensate and/or the second target condensate prior to determining the amount of test compound in the first condensate and/or the second condensate.

In some embodiments, dysregulation of the first target condensate and/or the second target condensate is associated with a disease. In some embodiments, the methods further comprise characterizing the first target condensate and/or the second target condensate by identifying one or more macromolecules comprised therein. In some embodiments, the methods further comprise labeling the first target condensate and/or the second target condensate in order to visualize the first target condensate and/or the second target condensate. In some embodiments, the methods further comprise labeling the first target condensate and the second target condensate in order to visualize the first condensate target condensate and the second target condensate. In some embodiments, the first target condensate and the second target condensate are labeled with different labels. In some embodiments, the first target condensate and/or the second target condensate are labeled with a radioactive label, a colorimetric label, a chemically-reactive label, or a fluorescent label.

In some embodiments, the composition comprises a cell. In some embodiments, the cell is a microorganism or an animal cell. In some embodiments, the cell comprises a condensate that is determined to be dysregulated. In some embodiments, the cell has one or more features of a neurodegenerative or proliferative disease.

In some embodiments, the first target condensate and/or the second target condensate are cellular condensates. In some embodiments, the first target condensate is a cleavage body, a P-granule, a histone locus body, a multivesicular body, a neuronal RNA granule, a nuclear gem, a nuclear pore, a nuclear speckle, a nuclear stress body, a nucleolus, a Oct1/PTF/transcription (OPT) domain, a paraspeckle, a perinucleolar compartment, a PML nuclear body, a PML oncogenic domain, a polycomb body, a processing body, a signaling cluster, a viral condensate, a Sam68 nuclear body, a stress granule, or a splicing speckle. In some embodiments, the second target condensate is a cleavage body, a P-granule, a histone locus body, a multivesicular body, a neuronal RNA granule, a nuclear gem, a nuclear pore, a nuclear speckle, a nuclear stress body, a nucleolus, a Oct1/PTF/transcription (OPT) domain, a paraspeckle, a perinucleolar compartment, a PML nuclear body, a PML oncogenic domain, a polycomb body, a processing body, a signaling cluster, a viral condensate, a Sam68 nuclear body, a stress granule, or a splicing speckle. In some embodiments, the first target condensate and/or the second target condensate are in a cell.

In some embodiments, the first target condensate and/or the second target condensate are extracellular condensates.

In some embodiments, the composition does not comprise a cell. In some embodiments, the composition comprises one or more of: a macromolecule, a salt, and a buffer.

In some embodiments, the composition comprises one or more additional target condensates. In some embodiments, the methods further comprise repeating the steps of the method for one or more additional target condensates.

In some aspects, provided herein are methods of determining condensate preference profiles of a plurality of test compounds, the method comprising performing a method of determining a condensate preference profile described herein with a plurality of test compounds. In some embodiments, the methods further comprise comparing condensate preference profiles of a subset or all of the plurality of test compounds. In some embodiments, the methods further comprise identifying test compounds that have the same or similar condensate preference profiles. In some embodiments, the methods further comprise identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar condensate preference profiles. In some embodiments, the methods further comprise determining the relative partition characteristic for one or more additional test compounds that comprise the identified characteristic. In some embodiments, the methods further comprise determining the relative partition characteristic for one or more additional test compounds that do not comprise the identified characteristic.

In some aspects, provided herein are methods of identifying a compound characteristic associated with partitioning a compound into or out of a condensate, the method comprising: (a) determining partition characteristics of a plurality of test compounds in the target condensate according to a method described herein; (b) comparing the partition characteristics of a subset or all of the plurality of test compounds in the target compound; (c) identifying test compounds that have the same or similar partition characteristics in the target condensate; and (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar partition characteristics.

In some aspects, provided herein are methods of identifying a compound characteristic associated with partitioning a compound into a condensate, the method comprising: (a) determining partition characteristics of a plurality of test compounds in the target condensate according to a method described herein; (b) comparing the partition characteristics of a subset or all of the plurality of test compounds in the target compound; (c) identifying test compounds that have the same or similar partition characteristics in the target condensate; and (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar partition characteristics.

In some aspects, provided herein are methods of identifying a compound characteristic associated with partitioning a compound into or out of a condensate, the method comprising: (a) determining relative partition characteristics of a plurality of test compounds in the target condensate according to a method described herein; (b) comparing the relative partition characteristics of a subset or all of the plurality of test compounds in the target condensate; (c) identifying test compounds that have the same or similar relative partition characteristics in the target condensate; and (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar relative partition characteristics.

In some aspects, provided herein are methods of identifying a compound characteristic associated with partitioning a compound into a condensate, the method comprising: (a) determining relative partition characteristics of a plurality of test compounds in the target condensate according to a method described herein; (b) comparing the relative partition characteristics of a subset or all of the plurality of test compounds in the target condensate; (c) identifying test compounds that have the same or similar relative partition characteristics in the target condensate; and (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar relative partition characteristics.

In some aspects, provided herein are methods of identifying a compound characteristic associated with partitioning a compound into or out of a condensate, the method comprising: (a) determining condensate preference profiles of a plurality of test compounds according to a method described herein; (b) comparing the condensate preference profiles of a subset or all of the plurality of test compounds; (c) identifying test compounds that have the same or similar condensate preference profiles; and (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar condensate preference profiles.

In some aspects, provided herein are methods of identifying a compound characteristic associated with partitioning a compound into a condensate, the method comprising: (a) determining condensate preference profiles of a plurality of test compounds according to a method described herein; (b) comparing the condensate preference profiles of a subset or all of the plurality of test compounds; (c) identifying test compounds that have the same or similar condensate preference profiles; and (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar condensate preference profiles.

In some aspects, provided herein are methods of designing a compound with a desired partition characteristic into or out of a target condensate, the method comprising: (a) determining partition characteristics of a plurality of test compounds in the target condensate according to a method described herein; (b) comparing the partition characteristics of a subset or all of the plurality of test compounds in the target condensate; (c) identifying test compounds that have the same or similar partition characteristics in the target condensate; (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar partition characteristics; and (e)(i) designing a compound that comprises the identified characteristic; or (ii) designing a compound that does not comprise the identified characteristic, thereby designing a compound with the desired partition characteristic into or out of the target condensate. In some embodiments, designing the compound comprises attaching a moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, designing the compound comprises removing a moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, designing the compound comprises changing a first moiety to a second moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, the compound is designed, in whole or in part, using an approach comprising a modeling, computer, and/or calculation-based technique, e.g., a bioinformatic, cheminformatic, and/or artificial intelligence (AI)-based technique.

In some aspects, provided herein are methods of designing a compound with a desired partition characteristic to a target condensate, the method comprising: (a) determining partition characteristics of a plurality of test compounds in the target condensate according to a method described herein; (b) comparing the partition characteristics of a subset or all of the plurality of test compounds in the target condensate; (c) identifying test compounds that have the same or similar partition characteristics in the target condensate; (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar partition characteristics; and (e) designing a compound that comprises the identified characteristic. In some embodiments, designing the compound comprises attaching a moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, designing the compound comprises changing a first moiety to a second moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, the compound is designed, in whole or in part, using an approach comprising a modeling, computer, and/or calculation-based technique, e.g., a bioinformatic, cheminformatic, and/or artificial intelligence (AI)-based technique.

In some aspects, provided herein are methods of designing a compound with a desired relative partition characteristic into or out of a target condensate, the method comprising: (a) determining relative partition characteristics of a plurality of test compounds in the target condensate according to a method disclosed herein; (b) comparing the relative partition characteristics of a subset or all of the plurality of test compounds in the target condensate; (c) identifying test compounds that have the same or similar relative partition characteristics in the target condensate; (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar relative partition characteristics; and (e)(i) designing a compound that comprises the identified characteristic; or (ii) designing a compound that does not comprise the identified characteristic, thereby designing a compound with the desired relative partition characteristic into or out of the target condensate. In some embodiments, designing the compound comprises attaching a moiety that comprises the identified characteristic, thereby conferring the desired relative partition characteristic to the compound. In some embodiments, designing the compound comprises removing a moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, designing the compound comprises changing a first moiety to a second moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, the compound is designed, in whole or in part, using an approach comprising a modeling, computer, and/or calculation-based technique, e.g., a bioinformatic, cheminformatic, and/or artificial intelligence (AI)-based technique.

In some aspects, provided herein are methods of designing a compound with a desired relative partition characteristic, the method comprising: (a) determining relative partition characteristics of a plurality of test compounds in the target condensate according to a method described herein; (b) comparing the relative partition characteristics of a subset or all of the plurality of test compounds in the target condensate; (c) identifying test compounds that have the same or similar relative partition characteristics in the target condensate; (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar relative partition characteristics; and (e) designing a compound that comprises the identified characteristic. In some embodiments, designing the compound comprises attaching a moiety that comprises the identified characteristic, thereby conferring the desired relative partition characteristic to the compound. In some embodiments, designing the compound comprises changing a first moiety to a second moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, the compound is designed, in whole or in part, using an approach comprising a modeling, computer, and/or calculation-based technique, e.g., a bioinformatic, cheminformatic, and/or artificial intelligence (AI)-based technique.

In some aspects, provided herein are methods of designing a compound with a desired condensate preference profile, the method comprising: (a) determining condensate preference profiles of a plurality of test compounds according to a method disclosed herein; (b) comparing the condensate preference profiles of a subset or all of the plurality of test compounds; (c) identifying test compounds that have the same or similar condensate preference profiles; (d) identifying a characteristic that a subset or all of the identified test compounds have in common in addition to the same or similar condensate preference profiles; and (e)(i) designing a compound that comprises the identified characteristic; or (ii) designing a compound that does not comprise the identified characteristic, thereby designing a compound with the desired condensate preference profile. In some embodiments, the methods further comprise making the compound. In some embodiments, designing the compound comprises attaching a moiety that comprises the identified characteristic, thereby conferring the desired condensate preference profile to the compound. In some embodiments, designing the compound comprises removing a moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, designing the compound comprises changing a first moiety to a second moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, the compound is designed, in whole or in part, using an approach comprising a modeling, computer, and/or calculation-based technique, e.g., a bioinformatic, cheminformatic, and/or artificial intelligence (AI)-based technique.

In some aspects, provided herein are methods of designing a compound with a desired condensate preference profile, the method comprising: (a) determining condensate preference profiles of a plurality of test compounds according to a method described herein; (b) comparing the condensate preference profiles of a subset or all of the plurality of test compounds; (c) identifying test compounds that have the same or similar condensate preference profiles; (d) identifying a characteristic, such as a chemical moiety or motif, that a subset or all of the identified test compounds have in common in addition to the same or similar condensate preference profiles; and (e) designing a compound that comprises the identified characteristic. In some embodiments, designing the compound comprises attaching a moiety that comprises the identified characteristic, thereby conferring the desired condensate preference profile to the compound. In some embodiments, designing the compound comprises changing a first moiety to a second moiety that comprises the identified characteristic, thereby conferring the desired partition characteristic to the compound. In some embodiments, there is provided a plurality of compounds designed by the methods described herein. In some embodiments, the compound is designed, in whole or in part, using an approach comprising a modeling, computer, and/or calculation-based technique, e.g., a bioinformatic, cheminformatic, and/or artificial intelligence (AI)-based technique. In some embodiments, the methods further comprise making the compound.

In some aspects, provided herein are methods of screening a test compound for a desired partition characteristic from a group of candidate compounds, the method comprising: (a) determining a partition characteristic of each of the group of candidate compounds; and (b) identifying the test compound having the desired partition characteristic. In some embodiments, the partition characteristic of each of the group of candidate compounds is determined in vitro. In some embodiments, the test compound has a suitable partition characteristic for being useful for treating a disease in an individual.

In some aspects, provided herein are methods of identifying a test compound useful for treating a disease in an individual in need thereof, the method comprising: (a) identifying a target condensate associated with the disease; and (b) determining a partition characteristic of a candidate compound in the target condensate, and (c) identifying the test compound having a suitable partition characteristic for being useful for treating the disease.

In some aspects, provided herein are methods of determining a partition characteristic of a test compound in a target condensate, the methods comprising: (a) combining the test compound and a composition comprising the target condensate and an extra-condensate solution; (b) obtaining a reference control; (c) measuring a MS signal of the test compound in the extra-condensate solution, or a portion thereof, using a mass spectrometry technique; (d) measuring a MS signal of the test compound in the reference control, or a portion thereof, using a mass spectrometry technique; and (e) comparing the MS signal of the test compound from the extra-condensate solution and the MS signal of the test compound from the reference control, thereby determining the partition characteristic of the test compound in the target condensate. In some embodiments, the amount of the test compound combined with the composition is 100 nM or less, and the amount of a precursor molecule in the composition, including in the target condensate, is about 5 μM.

In some aspects, provided herein is a library comprising a plurality of compounds, wherein each compound of the plurality of compounds comprises the same moiety comprising a characteristic having a desired partition characteristic.

In some aspects, provided herein is a method of designing a test compound having a desired partition characteristic, the method comprising modifying a precursor of the test compound by attaching a moiety to the compound, wherein the moiety comprises a characteristic having a desired partition characteristic.

It will also be understood by those skilled in the art that changes in the form and details of the implementations described herein may be made without departing from the scope of this disclosure. In addition, although various advantages, aspects, and objects have been described with reference to various implementations, the scope of this disclosure should not be limited by reference to such advantages, aspects, and objects.

