Hydroxyphenyl-indoline-2-one cancer therapeutics

Abstract

Small molecule ERα biomodulators that kill therapy-resistant ERα positive breast, ovarian, and endometrial cancer cells are disclosed. In one embodiment, the small molecule biomodulator has increased therapeutic utility because of an increased ability to kill therapy-resistant cancer cells compared to BHPI and other conventional therapies (endocrine therapies, tamoxifen, and fulvestrant / ICI). The small molecule biomodulators not only inhibit proliferation of the cancer cells but kills them, which prevents reactivation of tumors years later. Compounds of the invention, such as ErSO-TFPy, are effective for treating ERα positive cancers such as breast cancer, ovarian cancer, uterine cancer, cervical carcinoma, endometrial cancer, and the like.

Description

[0001] HYDR0XYPHENYL-IND0LINE-2-0NE CANCER THERAPEUTICS

[0002] RELATED APPLICATIONS

[0003] This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 63 / 682,128, filed August 12, 2024, which is incorporated herein by reference.

[0004] GOVERNMENT SUPPORT

[0005] This invention was made with government support under R01CA258746 awarded by the National Institutes of Health. The government has certain rights in the invention.

[0006] BACKGROUND OF THE INVENTION

[0007] While endocrine therapy has substantially improved the five-year survival rates for patients with ERa+ breast cancer, there is still an acute need for the treatment of drug resistant, advanced ERa+ breast cancers. There are numerous examples of new therapies seeking to address this need, including novel next-generation SERDs and ERa-targeting proteolysis targeting chimeras (PROTACs). However, these endocrine therapies utilize known inhibitory mechanisms that induce cancer cell cytostasis and lead to only limited tumor regression in preclinical models. This challenge is exemplified by the recently reported next-generation SERD, GDC-9545 (giredestrant) that has potent antiproliferative activity in cell culture but fails to quantitatively regress highly estrogen-dependent MCF-7 tumors, even in combination with CDK4 / 6 inhibitor palbociclib. The slow tumor regression induced by these drugs has implications for clinical applications, including the avoidance of using endocrine therapies in patients with large tumor burdens and significant toxicities upon long-term endocrine therapy treatment leading to the well-reported problem of poor patient compliance.

[0008] Targeting ERa directly or indirectly has been the flagship targeted strategy for treating ERa+ breast cancers. Such strategies are termed as endocrine therapies, as they seek to block the canonical functions of ERa in induction of cell growth and proliferation. Although patients initially benefit from endocrine therapy (typically with tamoxifen, fulvestrant, anastrozole, and others), drug-resistance and tumor regrowth is common and represents a major clinical challenge. Even with the recent combining of endocrine therapy with CDK4 / 6 or PI3KCA inhibitors, drug-resistant, lethal disease is still pervasive. These resistant tumors typically maintain their ERa overexpression, suggesting that drugs leveraging ERa, especially those acting through new mechanisms, may provide a meaningful clinical benefit to a diverse patient population that is typically considered drug resistant and is not well-served by other therapeutic options.

[0009] The traditional unfolded protein response (UPR) is characterized by signaling in response to insufficient protein-folding capacity. There is another form of the UPR, the anticipatory UPR (a-UPR), which is activated by cells in ‘anticipation’ of future hyperactivated growth. This a-UPR is a conserved mechanism across hormone-specific receptors including ERa, androgen receptor, epidermal growth factor receptor, and others; induction of the a-UPR can be considered as a non-canonical function of ERa. While the a-UPR is cytoprotective and has been correlated with drug-resistance in a variety of settings, sustained activation is toxic to cancer cells and represents an opportunity to convert a tumor protective pathway into a potent and selective anticancer strategy.

[0010] In contrast to the cytostatic activities of endocrine therapies, a-UPR activators have the potential to be highly effective, fast-acting, cytotoxic ERa-dependent therapies. ErSO is a member of the substituted 3-(4-hydroxyphenyl)indoline-2-one class of anticancer small molecules. A well-known member of this class is oxyphenisatin, a laxative used for over 40 years which later was shown to have antiproliferative effects against transformed and cancer cells. Derivatives of oxyphenisatin have potent anticancer activity. For example, Andruska et al (Proc. Natl. Acad. Sci. USA, 2015, 112 (15), 4737) disclosed BHPI, an oxyphenisatin derivative that retards the growth of ERa+ cancer cells through hyperactivation of ERa-mediated a-UPR. Initial demonstrations of the a-UPR’ s ‘draggability’ with BHPI was the basis for the discovery of ErSO, a compound that induces stark and unique cytotoxic a- UPR hyperactivation. While progenitor a-UPR activator BHPI is cytostatic against most ERa+ cancer cell lines, ErSO is rapidly cytotoxic and induces quantitative tumor regression in preclinical tumor models as a single agent with once-a-week dosing.

[0011] We recently reported the small molecule ErSO, a compound that hyperactivates the ERa- dependent a-UPR in ERa+ cancer cells leading to eradication of ERa+ breast tumors in multiple mouse models. ErSO maintains activity against breast cancer cell lines that contain mutations in ERa (Y537S and D538G) representing major clinical resistance mechanisms to endocrine therapy. The selectivity of ErSO for ERa+ cells in culture at short incubation times (e.g., 6 or 24 hours) is impressive, with >350 fold difference in cellular ICso between ERa+ and ERa- cancer cell lines. In addition, knock-in of ERa into ERa-negative MDA-MB-231 triple-negative breast cancer cells dramatically sensitizes these cells to ErSO both in cell culture and in xenograft models. The ability of ErSO to leverage this non-canonical activity of ERa (induction of the a-UPR) likely enables ErSO to induce tumor regressions, contrasting the cytostatic activity typically observed with endocrine therapy.

[0012] General lipophilicity (predicted by cLogD?.4) can have major implications for a compound’s selectivity and idiosyncratic toxicity profde. While lipophilicity can be critical for high affinity ligands and on-target mechanism of action, too much lipophilicity may lead to narrowed therapeutic windows driven by off-target mechanisms and other liabilities. ErSO’s poor lipophilic efficiency (LipE = 1.26) may play a role in the observed erosion of ERa-dependent activity in cell culture.

[0013] As previously reported with ErSO, 3-(4-hydroxyphenyl)indoline-2-ones can suffer from rat speciation toxicity. The exact underlying pharmacologic explanation and human relevance of this toxicity is not known. However, when considering tolerability the multi-year clinical use of oxyphenisatin is encouraging; while this drug was ultimately withdrawn from the clinic due to a rare hepatotoxicity that is untenable for a laxative drug, it was widely prescribed for four decades. Regardless, this rat toxicity can be used as a key preclinical filter for this drug class. Outside of simple cycloalkane substitutions, there has only been a limited set of substitutions to diversify 3 -(4- hydroxyphenyl)indoline-2-ones. To optimize lipophilic efficiencies, cancer cell line specificity, and rat in vivo tolerability, we developed a highly modular strategy to access a variety of diverse polar derivatives of this class.

[0014] The profound antitumor effects seen with a-UPR activators warrants their further investigation with a specific need to delineate direct target engagement and other aspects of their underlying mechanism. ErSO’s apparent ERa-independent effects, in some cancer cell lines when assessed at longer compound incubation times, may complicate studies surrounding target engagement, a-UPR activation, and cancer cell death.

[0015] Accordingly, new small molecule therapeutic agents that are cytotoxic, and not merely cytostatic, are urgently needed to provide more efficacious cancer therapy.

[0016] SUMMARY

[0017] Estrogen receptor alpha-positive (ERa+) breast cancers are the most common type of this disease, with >200,000 new cases diagnosed annually in the United States. For these cancers, ERa drives tumor growth and disease progression, and thus targeting ERa directly with ERa antagonists and degraders (e.g., tamoxifen, fulvestrant) or indirectly with aromatase inhibitors has been a successful therapeutic strategy, with significant gains in overall survival for these patients. However, such treatments are rarely curative, and patients typically succumb to metastatic, drug-resistant (through ERa mutation and other mechanisms) disease. The small molecule, ErSO, which induces potent ERa-dependent death of ERa+ breast cancer cells through a mechanism distinct from clinically approved drugs that target ERa, namely activation of the TRPM4 ion channel and subsequent cell swelling / endoplasmic reticulum stress. ErSO has remarkable activity in multiple mouse models of ERa+ breast cancer, in many cases inducing complete tumor eradication. Importantly, ErSO is potent and effective even when evaluated in breast cancer cell lines and preclinical tumor models that are resistant to endocrine therapy via mutated ERa. While ErSO has tremendous promise as a new drug to target ERa+ tumors, at high concentrations and long incubation times it does have some effects on ERa-negative (ERa-) cells in culture.

[0018] Fluorine atoms can have profound effects on a ligand’s physiochemical and confirmational properties and contribute to a variety of interactions between ligands and their protein target. Indeed, fluorination of nitrogen heterocycles utilized herein does decrease the predicted pAh of the corresponding conjugate acid and that p / / a perturbation may be a factor in their potent activity. Further, fluorination of piperidines (and other heterocycles) can have significant effects on compound conformation and may alter the dominate conformer in solution and / or in target binding. Herein we report the construction of modified versions of ErSO, with a major focus on establishing a structure-activity relationship and identifying new variants with an even wider differential activity between ERa+ and ERa- cells.

[0019] Accordingly, this disclosure provides a compound of Formula I: or a pharmaceutically acceptable salt thereof; wherein

[0020] W is CR1or N;

[0021] R1is halo, substituted -(C1-C6)alkyl, or unsubstituted or substituted -(C2-C8)alkynyl;

[0022] X is NRAor O;

[0023] RAIS H or -(Ci-C6)alkyl;

[0024] R2and R3are each independently H, halo, CN, NO2, or -(C1-C6)alkyl;

[0025] R4is H, -(C1-C6)alkyl, or -(C=O)(Ci-C6)alkyl;

[0026] Y is O or S;

[0027] Z1is substituted or unsubstituted 4-, 5- or 6-membered nitrogen heterocycle, substituted phenyl, substituted -NH(C2-C6)alkyl, or substituted -(C2-C8)alkynyl; and m is 0 or 1.

[0028] This disclosure also provides a method for treating cancer in a patient in need thereof, comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound described above and an excipient, wherein the cancer is thereby treated.

[0029] The invention provides novel compounds of Formulas I-III, intermediates for the synthesis of compounds of Formulas I-III, as well as methods of preparing compounds of Formulas I-III. The invention also provides compounds of Formulas I-III that are useful as intermediates for the synthesis of other useful compounds. The invention provides for the use of compounds of Formulas I-III for the manufacture of medicaments useful for the treatment of bacterial infections in a mammal, such as a human.

[0030] The invention provides for the use of the compositions described herein for use in medical therapy. The medical therapy can be treating cancer, including but not limited to breast cancer, lung cancer, pancreatic cancer, prostate cancer, or colon cancer. The invention also provides for the use of a composition as described herein for the manufacture of a medicament to treat a disease in a mammal, for example, cancer in a human. The medicament can include a pharmaceutically acceptable diluent, excipient, or carrier. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] The following drawings form part of the specification and are included to further demonstrate certain embodiments or various aspects of the invention. In some instances, embodiments of the invention can be best understood by referring to the accompanying drawings in combination with the detailed description presented herein. The description and accompanying drawings may highlight a certain specific example, or a certain aspect of the invention. However, one skilled in the art will understand that portions of the example or aspect may be used in combination with other examples or aspects of the invention.

[0032] Figure 1A-F. (A, B) Characterization of lead compounds. (C, D) Data showing breast cancer tumor regressions. (E, F) In-vitro data for compounds shown in Chart 2-4.

[0033] Figure 2. ErSO-DFP is the active anticancer agent with superior LipE relative to its progenitor, ErSO. (A) Biological activity of the racemic 2 ((±)-2), ErSO-DFP, and (X)-2 against MCF-7 cells incubated with compound for 24 h. Cell viability was assessed by alamar blue fluorescence. Raptinal (100 pM) was used as the 100 % dead control. Data is shown as mean ± s.e.m.; n > 2 independent replicates. (B) Lipophilic efficiencies (LipE) of ErSO and ErSO-DFP. LipE = pIC50 - cLogD7.4.

[0034] Figure 3A-B. Importance of Fluorine in 6-membered and 4-membered nitrogen heterocycles 16, ErSO-DFP, 35, and 25. Stability in PBS at 37 °C for 2 hours measured using LCMS, λ = 254 nm, (n > 2). Compounds were tested in MCF-7 cells for 24 hours and viability measured via alamar blue fluorescence (n > 2). Amine pKa and % protonated were calculated using ChemAxon MarvinSketch.

[0035] Figure 4. Evaluation of ErSO, ErSO-DFP, and ErSO-TFPy in hemolysis assay. Triton-X used as positive control (set to 100% hemolysis). 2 hr incubation at 37 °C followed by absorbance reading at 540 nm (n = 3).

[0036] Figure 5A-C. Cell culture activity of (A) ErSO, ErSO-DFP, and ErSO-TFPy against “normal” cell lines. Dose response curves of compounds against MCF-7 and “normal” cell lines. Compounds were tested for 72 hours and viability measured via alamar blue fluorescence (n > 2).

[0037] Figure 6. Stability of ErSO, ErSO-TFPy, and core derivatives in simulated gastric fluid (SGF) supplemented with pepsin (Chart 6). Compounds were incubated with SGF for 2 hours at 37 °C and concentrations were measured via LCMS, X = 254 nm, (n > 2).

[0038] DETAILED DESCRIPTION

[0039] Based on data from pre-clinical models, ErSO appears to have a wide therapeutic window; it is tolerated at oral doses greater than 150 mg / kg in mice and canines. However, there are traits of ErSO that, if altered, might lead to an even more promising drug. First, as compared to oral dosing, the maximum tolerated dose of ErSO in mice is significantly lower when administered intravenously (20 mg / kg), and there appears to be some species-specific sensitivity, with lower maximum tolerated doses in rats. Second, as shown and as detailed further herein, at longer incubation times and higher concentrations the selectivity of ErSO for induction of cell death in ERa+ versus ERa- cancer cells begins to erode in some cases. Herein we sought to construct and evaluate novel compounds related to ErSO, with the goal of identifying new leads with minimized ERa-independent effects; we hypothesized that such an optimized compound would have a wider therapeutic window in vivo. In addition to their translational promise, more selective compounds would also be superior probes for robust and clean ERa-dependent activation of the a-UPR.

[0040] Herein we report our efforts to synthesize 3-(4-hydroxyphenyl)indoline-2-one derivatives with improved selectivity and tolerability which has resulted in a number of novel compounds, a better understanding of structure-activity relationship surrounding the scaffold, and the discovery of several promising small molecules including ErSO-DFP and ErSO-TFPy. The lipophilicity and stability of these compounds appear to be key parameters determinant of antiproliferative activity and selectivity between cancer cell lines. Importantly, these derivatives are well tolerated across animal models and demonstrate impressive activity in preclinical models for breast cancer. ErSO-TFPy was found to be capable of inducing complete regressions in murine xenografts following a single dose, a very unusual and promising attribute for translational development.

[0041] This disclosure focuses on improvements in the activity of ErSO molecules (e.g., the 3-(4- hydroxyphenyl)indoline-2-one scaffold in breast cancer) through modifications to the phenyl rings and the indoline-2-one core (Chart 1).

[0042] Chart 1. Structure of ErSO and physicochemical parameters.

[0043] Physicochemical parameters calculated using ChemAxon MarvinSketch.

[0044] Additional information and data supporting the invention can be found in the following patent application publications: US 2023 / 0391721 and US 2021 / 0276951, which publications are incorporated herein by reference in their entirety. However, some embodiments of the compounds and formulas of this disclosure exclude specific compounds disclosed in the aforementioned patent application publications.

[0045] Definitions.

[0046] The following definitions are included to provide a clear and consistent understanding of the specification and claims. As used herein, the recited terms have the following meanings. All other terms and phrases used in this specification have their ordinary meanings as one of skill in the art would understand. Such ordinary meanings may be obtained by reference to technical dictionaries, such as Hawley ’s Condensed Chemical Dictionary 14thEdition, by R. J. Lewis, John Wiley & Sons, New York, N.Y., 2001.

[0047] References in the specification to "one embodiment", "an embodiment", etc., indicate that the embodiment described may include a particular aspect, feature, structure, moiety, or characteristic, but not every embodiment necessarily includes that aspect, feature, structure, moiety, or characteristic. Moreover, such phrases may, but do not necessarily, refer to the same embodiment referred to in other portions of the specification. Further, when a particular aspect, feature, structure, moiety, or characteristic is described in connection with an embodiment, it is within the knowledge of one skilled in the art to affect or connect such aspect, feature, structure, moiety, or characteristic with other embodiments, whether or not explicitly described.

[0048] The singular forms "a," "an," and "the" include plural reference unless the context clearly dictates otherwise. Thus, for example, a reference to "a compound" includes a plurality of such compounds, so that a compound X includes a plurality of compounds X. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for the use of exclusive terminology, such as "solely," "only," and the like, in connection with any element described herein, and / or the recitation of claim elements or use of "negative" limitations.

[0049] The term "and / or" means any one of the items, any combination of the items, or all of the items with which this term is associated. The phrases "one or more" and "at least one" are readily understood by one of skill in the art, particularly when read in context of its usage. For example, the phrase can mean one, two, three, four, five, six, ten, 100, or any upper limit approximately 10, 100, or 1000 times higher than a recited lower limit. For example, one or more substituents on a phenyl ring refers to one to five, or one to four, for example if the phenyl ring is disubstituted.

[0050] As will be understood by the skilled artisan, all numbers, including those expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth, are approximations and are understood as being optionally modified in all instances by the term "about." These values can vary depending upon the desired properties sought to be obtained by those skilled in the art utilizing the teachings of the descriptions herein. It is also understood that such values inherently contain variability resulting from the standard deviations found in their respective testing measurements. When values are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value without the modifier "about" also forms a further aspect.

[0051] The terms "about" and "approximately" are used interchangeably. Both terms can refer to a variation of ± 5%, ± 10%, ± 20%, or ± 25% of the value specified. For example, "about 50" percent can in some embodiments carry a variation from 45 to 55 percent, or as otherwise defined by a particular claim. For integer ranges, the term "about" can include one or two integers greater than and / or less than a recited integer at each end of the range. Unless indicated otherwise herein, the terms "about" and "approximately" are intended to include values, e.g., weight percentages, proximate to the recited range that are equivalent in terms of the functionality of the individual ingredient, composition, or embodiment. The terms "about" and "approximately" can also modify the endpoints of a recited range as discussed above in this paragraph.

[0052] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges recited herein also encompass any and all possible subranges and combinations of sub-ranges thereof, as well as the individual values making up the range, particularly integer values. It is therefore understood that each unit between two particular units are also disclosed. For example, if 10 to 15 is disclosed, then 11, 12, 13, and 14 are also disclosed, individually, and as part of a range. A recited range (e.g., weight percentages or carbon groups) includes each specific value, integer, decimal, or identity within the range. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, or tenths. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art, all language such as "up to", "at least", "greater than", "less than", "more than", "or more", and the like, include the number recited and such terms refer to ranges that can be subsequently broken down into sub-ranges as discussed above. In the same manner, all ratios recited herein also include all sub-ratios falling within the broader ratio. Accordingly, specific values recited for radicals, substituents, and ranges, are for illustration only; they do not exclude other defined values or other values within defined ranges for radicals and substituents. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint.

[0053] This disclosure provides ranges, limits, and deviations to variables such as volume, mass, percentages, ratios, etc. It is understood by an ordinary person skilled in the art that a range, such as “number 1” to “number?”, implies a continuous range of numbers that includes the whole numbers and fractional numbers. For example, 1 to 10 means 1, 2, 3, 4, 5, ... 9, 10. It also means 1.0, 1.1, 1.2. 1.3, ... , 9.8, 9.9, 10.0, and also means 1.01, 1.02, 1.03, and so on. If the variable disclosed is a number less than “numberlO”, it implies a continuous range that includes whole numbers and fractional numbers less than numberlO, as discussed above. Similarly, if the variable disclosed is a number greater than “numberlO”, it implies a continuous range that includes whole numbers and fractional numbers greater than numberlO. These ranges can be modified by the term “about”, whose meaning has been described above.

[0054] One skilled in the art will also readily recognize that where members are grouped together in a common manner, such as in a Markush group, the invention encompasses not only the entire group listed as a whole, but each member of the group individually and all possible subgroups of the main group. Additionally, for all purposes, the invention encompasses not only the main group, but also the main group absent one or more of the group members. The invention therefore envisages the explicit exclusion of any one or more of members of a recited group. Accordingly, provisos may apply to any of the disclosed categories or embodiments whereby any one or more of the recited elements, species, or embodiments, may be excluded from such categories or embodiments, for example, for use in an explicit negative limitation.

[0055] The term "contacting" refers to the act of touching, making contact, or of bringing to immediate or close proximity, including at the cellular or molecular level, for example, to bring about a physiological reaction, a chemical reaction, or a physical change, e.g., in a solution, in a reaction mixture, in vitro, or in vivo.

[0056] An "effective amount" refers to an amount effective to treat a disease, disorder, and / or condition, or to bring about a recited effect. For example, an effective amount can be an amount effective to reduce the progression or severity of the condition or symptoms being treated. Determination of a therapeutically effective amount is well within the capacity of persons skilled in the art. The term "effective amount" is intended to include an amount of a compound described herein, or an amount of a combination of compounds described herein, e.g., that is effective to treat or prevent a disease or disorder, or to treat the symptoms of the disease or disorder, in a host. Thus, an "effective amount" generally means an amount that provides the desired effect.

[0057] Alternatively, the terms "effective amount" or "therapeutically effective amount," as used herein, refer to a sufficient amount of an agent or a composition or combination of compositions being administered which will relieve to some extent one or more of the symptoms of the disease or condition being treated. The result can be reduction and / or alleviation of the signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic uses is the amount of the composition comprising a compound as disclosed herein required to provide a clinically significant decrease in disease symptoms. An appropriate "effective" amount in any individual case may be determined using techniques, such as a dose escalation study. The dose could be administered in one or more administrations. However, the precise determination of what would be considered an effective dose may be based on factors individual to each patient, including, but not limited to, the patient's age, size, type or extent of disease, stage of the disease, route of administration of the compositions, the type or extent of supplemental therapy used, ongoing disease process and type of treatment desired (e.g., aggressive vs. conventional treatment).

[0058] The terms "treating", "treat" and "treatment" include (i) preventing a disease, pathologic or medical condition from occurring (e.g., prophylaxis); (ii) inhibiting the disease, pathologic or medical condition or arresting its development; (iii) relieving the disease, pathologic or medical condition; and / or (iv) diminishing symptoms associated with the disease, pathologic or medical condition. Thus, the terms "treat", "treatment", and "treating" can extend to prophylaxis and can include prevent, prevention, preventing, lowering, stopping or reversing the progression or severity of the condition or symptoms being treated. As such, the term "treatment" can include medical, therapeutic, and / or prophylactic administration, as appropriate.

[0059] As used herein, "subject" or “patient” means an individual having symptoms of, or at risk for, a disease or other malignancy. A patient may be human or non-human and may include, for example, animal strains or species used as “model systems” for research purposes, such a mouse model as described herein. Likewise, the patient may include either adults or juveniles (e.g., children). Moreover, patient may mean any living organism, preferably a mammal (e.g., human or non-human) that may benefit from the administration of compositions contemplated herein. Examples of mammals include, but are not limited to, any member of the Mammalian class: humans, non-human primates such as chimpanzees, and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice and guinea pigs, and the like. Examples of non-mammals include, but are not limited to, birds, fish and the like. In one embodiment of the methods provided herein, the mammal is a human.

[0060] As used herein, the terms “providing”, “administering,” “introducing,” are used interchangeably herein and refer to the placement of a compound of the disclosure into a subject by a method or route that results in at least partial localization of the compound to a desired site. The compound can be administered by any appropriate route that results in delivery to a desired location in the subject.

[0061] The compound and compositions described herein may be administered with additional compositions to prolong stability and activity of the compositions, or in combination with other therapeutic drugs.

[0062] The terms "inhibit", "inhibiting", and "inhibition" refer to the slowing, halting, or reversing the growth or progression of a disease, infection, condition, or group of cells. The inhibition can be greater than about 20%, 40%, 60%, 80%, 90%, 95%, or 99%, for example, compared to the growth or progression that occurs in the absence of the treatment or contacting.

[0063] The term “substantially” as used herein, is a broad term and is used in its ordinary sense, including, without limitation, being largely but not necessarily wholly that which is specified. For example, the term could refer to a numerical value that may not be 100% the full numerical value. The full numerical value may be less by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, or about 20%.

[0064] Wherever the term “comprising” is used herein, options are contemplated wherein the terms “consisting of’ or “consisting essentially of’ are used instead. As used herein, “comprising” is synonymous with "including," "containing," or "characterized by," and is inclusive or open-ended and does not exclude additional, unrecited elements or method steps. As used herein, "consisting of' excludes any element, step, or ingredient not specified in the aspect element. As used herein, "consisting essentially of' does not exclude materials or steps that do not materially affect the basic and novel characteristics of the aspect. In each instance herein any of the terms "comprising", "consisting essentially of' and "consisting of' may be replaced with either of the other two terms. The disclosure illustratively described herein may be suitably practiced in the absence of any element or elements, limitation or limitations which is not specifically disclosed herein.

[0065] This disclosure provides methods of making the compounds and compositions of the invention. The compounds and compositions can be prepared by any of the applicable techniques described herein, optionally in combination with standard techniques of organic synthesis. Many techniques such as etherification and esterification are well known in the art. However, many of these techniques are elaborated in Compendium of Organic Synthetic Methods (John Wiley & Sons, New York), Vol. 1, Ian T. Harrison and Shuyen Harrison, 1971; Vol. 2, Ian T. Harrison and Shuyen Harrison, 1974; Vol. 3, Louis S. Hegedus and Leroy Wade, 1977; Vol. 4, Leroy G. Wade, Jr., 1980; Vol. 5, Leroy G. Wade, Jr., 1984; and Vol. 6; as well as standard organic reference texts such as March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th Ed., by M. B. Smith and J. March (John Wiley & Sons, New York, 2001); Comprehensive Organic Synthesis. Selectivity, Strategy & Efficiency in Modem Organic Chemistry. In 9 Volumes, Barry M. Trost, Editor-in-Chief (Pergamon Press, New York, 1993 printing); Advanced Organic Chemistry, Part B: Reactions and Synthesis, Second Edition, Cary and Sundberg (1983); for heterocyclic synthesis see Hermanson, Greg T., Bioconjugate Techniques, Third Edition, Academic Press, 2013.

[0066] The formulas and compounds described herein can be modified using protecting groups. Suitable amino and carboxy protecting groups are known to those skilled in the art (see for example, Protecting Groups in Organic Synthesis, Second Edition, Greene, T. W., and Wuts, P. G. M., John Wiley & Sons, New York, and references cited therein; Philip J. Kocienski; Protecting Groups (Georg Thieme Verlag Stuttgart, New York, 1994), and references cited therein); and Comprehensive Organic Transformations, Larock, R. C., Second Edition, John Wiley & Sons, New York (1999), and referenced cited therein.

[0067] The term "halo" or "halide" refers to fluoro, chloro, bromo, or iodo. Similarly, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0068] The term "alkyl" refers to a branched or unbranched hydrocarbon having, for example, from 1- 20 carbon atoms, and often 1-12, 1-10, 1-8, 1-6, or 1-4 carbon atoms; or for example, a range between 1-20 carbon atoms, such as 2-6, 3-6, 2-8, or 3-8 carbon atoms. As used herein, the term “alkyl” also encompasses a “cycloalkyl”, defined below. Examples include, but are not limited to, methyl, ethyl, 1- propyl, 2-propyl ( / .so-propyl), 1 -butyl, 2-methyl-l -propyl (isobutyl), 2-butyl (sec-butyl), 2-methyl-2- propyl (Lbutyl), 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-l-butyl, 2- methyl-1 -butyl, 1 -hexyl, 2-hexyl, 3 -hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3 -methyl-3 -pentyl, 2-methyl-3 -pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, hexyl, octyl, decyl, dodecyl, and the like. The alkyl can be unsubstituted or substituted, for example, with a substituent described below or otherwise described herein. The alkyl can also be optionally partially or fully unsaturated. As such, the recitation of an alkyl group can include an alkenyl group or an alkynyl group. The alkyl can be a monovalent hydrocarbon radical, as described and exemplified above, or it can be a divalent hydrocarbon radical (i.e., an alkylene).

[0069] An alkylene is an alkyl group having two free valences at a carbon atom or two different carbon atoms of a carbon chain. Similarly, alkenylene and alkynylene are respectively an alkene and an alkyne having two free valences at two different carbon atoms, or an alkenylene can have the two free valences on the same carbon.

[0070] The term "cycloalkyl" refers to cyclic alkyl groups of, for example, from 3 to 10 carbon atoms having a single cyclic ring or multiple condensed rings. Cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, and the like, or multiple ring structures such as adamantyl, and the like. The cycloalkyl can be unsubstituted or substituted. The cycloalkyl group can be monovalent or divalent and can be optionally substituted as described for alkyl groups. The cycloalkyl group can optionally include one or more cites of unsaturation, for example, the cycloalkyl group can include one or more carbon-carbon double bonds, such as, for example, 1- cyclopent-l-enyl, 1 -cyclopent-2-enyl, 1 -cyclopent-3 -enyl, cyclohexyl, 1 -cyclohex- 1-enyl, 1-cyclohex- 2-enyl, 1 -cyclohex-3 -enyl, and the like.

[0071] The term “heteroatom” refers to any atom in the periodic table that is not carbon or hydrogen. Typically, a heteroatom is O, S, N, P. The heteroatom may also be a halogen, metal or metalloid.

[0072] The term "heterocycloalkyl" or “heterocyclyl” refers to a saturated or partially saturated monocyclic, bicyclic, or polycyclic ring containing at least one heteroatom selected from nitrogen, sulfur, oxygen, preferably from 1 to 3 heteroatoms in at least one ring. Each ring is preferably from 3- to 10-membered, more preferably 4 to 7 membered. Examples of suitable heterocycloalkyl substituents include pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiofuranyl, piperidinyl, piperazinyl, tetrahydropyranyl, morpholinyl, 1,3-diazapanyl, 1,4-diazapanyl, 1 ,4-oxazepanyl, and 1,4- oxathiapanyl. The group may be a terminal group or a bridging group.

[0073] The term "aryl" refers to an aromatic hydrocarbon group derived from the removal of at least one hydrogen atom from a single carbon atom of a parent aromatic ring system. The radical attachment site can be at a saturated or unsaturated carbon atom of the parent ring system. The aryl group can have from 6 to 30 carbon atoms, for example, about 6-10 carbon atoms. The aryl group can have a single ring (e.g., phenyl) or multiple condensed (fused) rings, wherein at least one ring is aromatic (e.g., naphthyl, dihydrophenanthrenyl, fluorenyl, or anthryl). Typical aryl groups include, but are not limited to, radicals derived from benzene, naphthalene, anthracene, biphenyl, and the like. The aryl can be unsubstituted or optionally substituted with a substituent described below. For example, a phenyl moiety or group may be substituted with one or more substituents Rxwhere Rxis at the ortho-, meta-, or para-position, and X is an integer variable of 1 to 5.

[0074] The term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic ring system containing one, two, or three aromatic rings and containing at least one nitrogen, oxygen, or sulfur atom in an aromatic ring. The heteroaryl can be unsubstituted or substituted, for example, with one or more, and in particular one to three, substituents, as described in the definition of "substituted". Typical heteroaryl groups contain 2-20 carbon atoms in the ring skeleton in addition to the one or more heteroatoms, wherein the ring skeleton comprises a 5-membered ring, a 6-membered ring, two 5-membered rings, two 6- membered rings, or a 5-membered ring fused to a 6-membered ring. Examples of heteroaryl groups include, but are not limited to, 2H-pyrrolyl, 3H-indolyl, 4H-quinolizinyl, acridinyl, benzo[b]thienyl, benzothiazolyl, 0-carbolinyl, carbazolyl, chromenyl, cinnolinyl, dibenzo[b,d]furanyl, furazanyl, furyl, imidazolyl, imidizolyl, indazolyl, indolisinyl, indolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphthyridinyl, oxazolyl, perimidinyl, phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, thiadiazolyl, thianthrenyl, thiazolyl, thienyl, triazolyl, tetrazolyl, and xanthenyl. In one embodiment the term "heteroaryl" denotes a monocyclic aromatic ring containing five or six ring atoms containing carbon and 1, 2, 3, or 4 heteroatoms independently selected from non-peroxide oxygen, sulfur, and N(Z) wherein Z is absent or is H, O, alkyl, aryl, or (Ci-C6)alkylaryl. In some embodiments, heteroaryl denotes an ortho-fused bicyclic heterocycle of about eight to ten ring atoms derived therefrom, particularly a benzo-derivative or one derived by fusing a propylene, trimethylene, or tetramethylene diradical thereto.

[0075] As used herein, the term "substituted" or “substituent” is intended to indicate that one or more (for example, in various embodiments, 1-10; in other embodiments, 1-6; in some embodiments 1, 2, 3, 4, or 5; in certain embodiments, 1, 2, or 3; and in other embodiments, 1 or 2) hydrogens on the group indicated in the expression using “substituted” (or “substituent”) is replaced with a selection from the indicated group(s), or with a suitable group known to those of skill in the art, provided that the indicated atom’s normal valency is not exceeded, and that the substitution results in a stable compound. Suitable indicated groups include, e.g., alkyl, alkenyl, alkynyl, alkoxy, haloalkyl, hydroxyalkyl, aryl, heteroaryl, heterocyclyl, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, alkylamino, dialkylamino, carboxyalkyl, alkylthio, alkylsulfinyl, and alkylsulfonyl. Substituents of the indicated groups can be those recited in a specific list of substituents described herein, or as one of skill in the art would recognize, can be one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, halo, haloalkyl, hydroxy, hydroxyalkyl, aryl, heteroaryl, heterocycle, cycloalkyl, alkanoyl, alkoxycarbonyl, amino, alkylamino, dialkylamino, trifluoromethylthio, difluoromethyl, acylamino, nitro, trifluoromethyl, trifluoromethoxy, carboxy, carboxyalkyl, keto, thioxo, alkylthio, alkylsulfinyl, alkylsulfonyl, and cyano. Suitable substituents of indicated groups can be bonded to a substituted carbon atom include F, Cl, Br, I, OR', 0C(0)N(R')2, CN, CF3, OCF3, R', O, S, C(O), S(O), methylenedioxy, ethylenedioxy, N(R')2, SR', SOR', SO2R', SO2N(R')2, SO3R', C(O)R', C(O)C(O)R', C(O)CH2C(O)R', C(S)R', C(O)OR', OC(O)R', C(O)N(R')2, OC(O)N(R')2, C(S)N(R')2, (CH2)O-2NHC(0)R', N(R')N(R')C(O)R', N(R')N(R')C(O)OR', N(R')N(R')CON(R')2, N(R')SO2R', N(R')SO2N(R')2, N(R')C(O)OR',

[0076] N(R')C(O)R', N(R')C(S)R', N(R')C(O)N(R')2, N(R')C(S)N(R')2, N(COR')COR', N(OR')R', C(=NH)N(R')2, C(O)N(OR')R', or C(=NOR')R' wherein R’ can be hydrogen or a carbon-based moiety (e.g., (C1-C6)alkyl), and wherein the carbon-based moiety can itself be further substituted. When a substituent is monovalent, such as, for example, F or Cl, it is bonded to the atom it is substituting by a single bond. When a substituent is divalent, such as O, it is bonded to the atom it is substituting by a double bond; for example, a carbon atom substituted with O forms a carbonyl group, C=O.

[0077] Stereochemical definitions and conventions used herein generally follow S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., “Stereochemistry of Organic Compounds”, John Wiley & Sons, Inc., New York, 1994. The compounds of the invention may contain asymmetric or chiral centers, and therefore exist in different stereoisomeric forms. It is intended that all stereoisomeric forms of the compounds of the invention, including but not limited to, diastereomers, enantiomers and atropisomers, as well as mixtures thereof, such as racemic mixtures, which form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane- polarized light. In describing an optically active compound, the prefixes D and L, or R and S. are used to denote the absolute configuration of the molecule about its chiral center(s). The prefixes d and 1 or (+) and (-) are employed to designate the sign of rotation of plane-polarized light by the compound, with (-) or 1 meaning that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of one another. A specific stereoisomer may also be referred to as an enantiomer, and a mixture of such isomers is often called an enantiomeric mixture. A 50:50 mixture of enantiomers is referred to as a racemic mixture or a racemate (defined below), which may occur where there has been no stereoselection or stereospecificity in a chemical reaction or process.

[0078] The terms “racemic mixture” and “racemate” refer to an equimolar mixture of two enantiomeric species, devoid of optical activity.

[0079] The term “enantiomerically enriched” (“ee”) as used herein refers to mixtures that have one enantiomer present to a greater extent than another. Reactions that provide one enantiomer present to a greater extent than another would therefore be “enantioselective” (or demonstrate “enantioselectivity”). In one embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 2% ee; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 5% ee; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 20%; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 50%; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 80%; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 90%; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 95%; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 98%; in another embodiment of the invention, the term “enantiomerically enriched” refers to a mixture having at least about 99%. The term “enantiomerically enriched” includes enantiomerically pure mixtures which are mixtures that are substantially free of the species of the opposite optical activity or one enantiomer is present in very low quantities, for example, 0.01%, 0.001% or 0.0001%.

[0080] The term “IC50” is generally defined as the concentration required to inhibit a specific biological or biochemical function by half, or to kill 50% of the cells in a designated time period, typically 24 hours.

[0081] Embodiments of the Technology.

[0082] 1. A compound of Formula I: or a pharmaceutically acceptable salt thereof; wherein

[0083] W is CR1or N;

[0084] R1is halo, substituted -(C1-C6)alkyl, or unsubstituted or substituted -(C2-C8)alkynyl;

[0085] X is NRAor O;

[0086] RAIS H or -(C1-C6)alkyl;

[0087] R2and R3are each independently H, halo, CN, NO2, or -(C1-C6)alkyl;

[0088] R4is H, -(C1-C6)alkyl, or -(C=O)(C1-C6)alkyl;

[0089] Y is O or S;

[0090] Z1is substituted or unsubstituted 4-, 5- or 6-membered nitrogen heterocycle, substituted phenyl, substituted -NH(C2-C6)alkyl, or substituted -(C2-C8)alkynyl; and m is 0 or 1.

[0091] In some embodiments, the substituent on the substituted 4-, 5- or 6-membered nitrogen heterocycle, substituted -NH(C2-C6)alkyl, or substituted -(C2-C8)alkynyl is a diazirine substituent, m is 0 or 1. In some embodiments, the substituent on the substituted -(C1-C6)alkyl is one or more fluoro, such as in CF3. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, when Z1is substituted 5 -or 6-membered nitrogen containing heterocycle and W is CR1, R1is not substituted -(Ci-C6)alkyl.

[0092] 2. The compound of embodiment 1, wherein Z1is substituted or unsubstituted pyrrolidine- 1-yl, pyrrolidine-3 -yl, piperidine- 1-yl, or piperidine-4-yl.

[0093] 3. The compound of embodiment 1, wherein Z1is:

[0094] The compound of embodiment 1, wherein Z1is: , wherein

[0095] R5is -O(C2-C8)alkynyl, N3, or nitrogen-substituted -OCH2-triazole; and

[0096] R6is H, -(C2-C8)alkynyl, -O(C2-C8)alkynyl, or nitrogen-substituted -OCH2-triazole;

[0097] The compound of embodiment 4, wherein one of R5or R6is nitrogen-substituted -OCH2- triazole and the triazole moiety of R6is: wherein n is 1-5 and p is 1-5.

[0098] 6. The compound of any one of embodiments 1-5, wherein m is 0.

[0099] 7. The compound of any one of embodiments 1-6, wherein X is NH and Y is O; or W is CR1; or W is CR1, X is NH, and Y is O.

[0100] In a preferred embodiment, the compound of Formula I is represented by Formula II: or a pharmaceutically acceptable salt thereof. In some other embodiment, the compound of Formula I is represented by Formula III: or a pharmaceutically acceptable salt thereof.

[0101] In some embodiments, compounds of Formula I, II, or III at position-3 of the indoIone heterocycle have a stereochemical (S)-configuration. In some other embodiments, compounds of Formula I or II at position-3 of the indoIone heterocycle have a stereochemical (R)-configuration.

[0102] 8. The compound of embodiment 1, wherein the compound is: or a pharmaceutically acceptable salt thereof.

[0103] 9. The compound of embodiment 1, wherein the compound is:

[0104] 10. The compound of embodiment 1, wherein the compound is: or a pharmaceutically acceptable salt thereof.

[0105] 11. A pharmaceutical composition comprising a compound of any one of embodiments 1-10 and a pharmaceutically acceptable excipient.

[0106] 12. A method for treating cancer in a patient in need thereof, comprising administering a therapeutically effective amount of a pharmaceutical composition according to embodiment 11, wherein the cancer is thereby treated.

[0107] 13. The method of embodiment 12, wherein the compound in the pharmaceutical composition is: or a pharmaceutically acceptable salt thereof.

[0108] 14. The method of embodiment 12 or 13, wherein the cancer is breast cancer.

[0109] 15. The method of any one of embodiments 12-14, wherein the therapeutically effective amount of the pharmaceutical composition administered comprises about 2 mg / kg (mg of drug per kg body wt.) to about 50 mg / kg of the compound. In some other embodiments, the mg / kg amount of compound in the composition administered is about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, or 200 mg / kg.

[0110] Results and Discussion.

[0111] Synthesis and Evaluation of 3-(4-hydroxyphenyl)-indoline-2-ones in Breast Cancer. Initial derivatives of BHPI and ErSO were prepared according to Table 1. Compounds shown in Charts 2- 4 were prepared (Scheme 1) and tested (Figure 1A-1F). They led to the discovery of ErSO-TFPy.

[0112]

[0113] Chart 2. Electron withdrawing group modification of core. Chart 4. Derivatives synthesized to interrogate binding interactions with target. IC50 Collected in

[0114] MCF-7 at 24 / 72 hr.

[0115] A. Noncovalent interactions.

[0116] B. Covalent interactions.

[0117]

[0118] Structure-Activity Relationships (A-Ring Modifications). Derivatives were synthesized with modifications to the A-Ring phenol. Access to the racemic mixtures of these derivatives can be obtained using the synthetic route in Scheme 2. Briefly, the B-ring connection is obtained by performing lithium-halogen exchange on l-bromo-4-(trifluoromethoxy)benzene followed by nucleophilic attack on the aromatic ketone of 7-trifluoromethyl isatin to afford the tertiary alcohol. Under strong acidic conditions, this alcohol will leave forming a stabilized tertiary carbocation, and the A-ring derivatives can be attached in Friedel-Crafts type alkylation. Racemic mixtures of derivatives of interest may then be separated via chiral chromatography to obtain the enantiomerically-pure compound. The racemate is expected to have half the biological activity of the corresponding active enantiomer; this is an important consideration when making comparisons between mixtures and enantiopure compounds.

[0119] Scheme 2. Modular Synthesis of Phenol Derivatives.

[0120] Reagents and conditions: (a) n-butyllithium, tetrahydrofuran, -78 °C to room temperature, 1 h, 65 %; (b) triflic acid, dichloromethane, 0 °C, 1 h 36-39 %. The objectives in derivatizing the phenol were to explore SAR, decrease lipophilicity of the scaffold, and confirm the importance of the phenol. Phenols are ubiquitous motifs in nature (e.g. tyrosine, serotonin) as well as drug development, but they are associated with poor oral bioavailability, metabolic instability, and toxicity concerns. As such, the main metabolite of ErSO is glucuronidation of the phenol, yet it has been shown to still maintain good oral bioavailability. Lipophilicity is an important parameter in drug development that affects potency / protein binding, clearance, and off- targets / toxicity. ErSO has high lipophilicity with a cLogD7.4 value of 6.4; values in this range are typically associated with promiscuity, thus one of the goals was to synthesize derivatives with decreased lipophilicity while maintaining cellular potency.

[0121] Table 1. Parameters and Cellular Potencies for A-Ring Derivatives. a:pKaand cLogD?.4 were calculated using ChemAxon MarvinSketch.b:pKais reported for Ring A acidic protons.c: To determine ICso, MCF-7 cells were incubated with compound for 24 hours and viability measured via alamar blue fluorescence. Raptinal (100 pM) was used as the 100 % dead control. Data is shown as mean%fc s.e.m.; n > 2 independent replicates. iPr: isopropyl. Note: ErSO-OH absolute configuration is inferred from ErSO structure.

[0122] Structures, cellular activity, and calculated parameters (phenolic pKa, cLogD7.4) are reported in Table 1. Replacement of the phenol with trifluoromethyl anisole (Compound 9) did result in a loss of activity in MCF-7 cells, confirming the importance of the phenol. Additionally, alkylation at the -ortho and -meta position with methyl (Compounds 11 & 12) and isopropyl groups (Compound 13) resulted in a complete loss of activity. A crystal structure of acetylated ErSO has previously been reported9and indicates that the A and B ring sit orthogonally to each other, which suggests that larger alkyl groups impede the free rotation of these two rings. Mono- and Di-fluorination at various positions (Compounds 5-8) result in a range of activities and significantly affect the calculated pKaof the phenol. In particular, Compounds 6 and 8 increase the acidity of the phenol and result in significant loss of anticancer activity, suggesting ideal pKavalues of 8-10. The catechol derivative (Compound 10) was one of the only derivatives to maintain significant potency while decreasing lipophilicity (cLogD?.4 = 6.1), so the enantiomers were separated via chiral chromatography to afford the derivative of interest ErSO-OH or (5)-10. ErSO-OH intravenous tolerability was assessed in CD-I mice and Sprague Dawley rats. ErSO- OH was tolerated at 25 mg / kg in mice but was lethal to rats at 15 mg / kg. Ultimately, this derivative was not pursued further as catechols are known to oxidize to the quinone, leading to promiscuous reactivity.

[0123] Structure-Activity Relationship (B-Ring Modifications) .

[0124] Saturated Nitrogen Heterocycles. Given the necessity of the phenol for activity and subsequent difficulties with introducing alterations into the A-Ring, subsequent efforts to explore SAR and decrease lipophilicity focused on the B-Ring. Previous studies have demonstrated that the B-ring can tolerate a greater diversity of derivatives, though this was limited primarily to substituted aromatic rings and unsubstituted cyclo-alkanes. To address the high lipophilicity of ErSO, efforts were focused on incorporating nitrogen into the B-ring. Efforts to derivatize with aromatic nitrogen heterocycles were unsuccessful, but saturated nitrogen heterocycles could be incorporated by rerouting through an unstable tertiary chloride (Scheme 3). The exact mechanism of nucleophilic attack on the tertiary chloride has not been studied, but we posit that the tertiary chloride (15) is less stable than the initial tertiary alcohol (14) and nitrogen nucleophiles may attack through either an SNi or possibly an SNi mechanism. This chemistry had been previously reported in the synthesis of alkyl aryl ethers and 3-N- aryl derivatives, but it had not been used for saturated nitrogen heterocycles or in the context of the anticancer activity of this scaffold. Dr. Matthew Boudreau was the first to apply this synthetic modification in this context and this route has offered access to greater diversity at this position as previous studies had primarily focused on bis-aryl scaffolds.

[0125] Scheme 3. Modular synthesis of B-Ring Derivatives3. aReagents and conditions: n-butyllithium, tetrahydrofuran, -78 °C to room temperature, 1 h, 79%; (b) thionyl chloride, pyridine, di chloromethane, 0 °C, 1 h, not isolated; (c) pyridine (if HO salt of amine nucleophile is used), tetrahydrofuran or dimethylformamide, room temperature, 2 -4 h, not isolated; (d) tetra-n-butylammonium fluoride, tetrahydrofuran, room temperature, 1-2 h, 41—83% (over three steps); TB S : tert-butyldimethylsilyl.

[0126] B-Ring derivatives and activity in MCF-7 cells are reported in Table 2. Compounds 2 and 16- 21 possessed lowered cLogD7.4 values ranging from 2.6 to 4.9 and amine pKavalues ranging from 3.6 to 6.2, suggesting that these compounds primarily exist as the neutral species at physiological pH (7.4). The difluorinated piperidine (DFP) derivative, Compound 2, had the most potent activity in MCF-7 cells (24 h IC50 = 35 nM) and a reduced cLogD7.4 value of 4.4 (relative to ErSO). Interestingly, Compounds 17 and 21 also bore fluorinated nitrogen heterocycles and maintained nanomolar potency while all the non-fluorinated compounds (16, 18-20) did not reach 50% cell death at the highest concentration tested (1 pM). Compound 17 was tested as a mixture of diastereomers, and its retention of activity indicated that branching off these fluorinated nitrogen heterocycles may be tolerated. Importantly, these studies highlighted the flexibility of the B-Ring but also raised questions regarding the importance of fluorination. Table 2. Parameters and Cellular Potencies for B-Ring Derivatives. apKa was calculated using ChemAxon MarvinSketch.bAmine pKa(i.e., tertiary amine on the final compound) was calculated using ChemAxon MarvinSketch.cTo determine ICso, MCF-7 cells were incubated with compound for 24 hours and viability measured via alamar blue fluorescence; raptinal (100 pM) was used as the 100% dead control; data is shown as mean ± s.e.m.; n > 2 independent replicates.dMixture of diastereomers and their corresponding enantiomers, 4 total compounds.

[0127] Compound 2 was selected for further investigation due to its potency and decreased lipophilicity. Enantiomers were separated via chiral chromatography and absolute stereochemistry obtained via x-ray crystallography (Figure 2A). Similar to previous derivatives, the active enantiomer, denoted ErSO-DFP, has the A-ring phenol oriented backwards and the B-ring oriented forwards. ErSO- DFP was found to have an IC50 of 17 nM in MCF-7 cells, which is expectantly twice the potency of the racemic mixture (±)-2. (A’)-2, the inactive enantiomer, had no activity when tested up to 1 pM, consistent with the reported differential activity of the enantiomers. Lipophilic efficiency, LipE, is an important parameter used to simultaneously track potency / lipophilicity in medicinal chemistry campaigns and is calculated using the following equation: LipE = -log(potency) - logD. The decreased lipophilicity of ErSO-DFP results in a higher LipE value of 3.4 compared to ErSO (1.3). More importantly, in a more extensive cancer cell line panel (Table 3), ErSO-DFP maintains comparable activity in ERa+ cell lines (T47D, TYS, TDG) and exhibited significantly less activity in ERa- cell lines (MDA-MB-231, MDA-MB-436, HCT-116, HT-29) than ErSO. This improved cell-line selectivity is important for translation and highlights the value in tuning lipophilicity of this scaffold. Furthermore, ErSO-DFP demonstrated improved tolerability in animal models (Mouse MTDiv = 95 mg / kg and Rat MTDiv >50 mg / kg) and induced tumor regressions in an MCF-7 xenograft.

[0128] Table 3. Cell Culture Activity for ErSO and ErSO-DFP.

[0129] ErSO ErSO-DFP a:To determine IC50, cancer cells were incubated with compound for 24, 72, or 168 h and viability was measured via alamar blue fluorescence; raptinal (100 pM) was used as the 100% dead control; data is shown as mean ± s.e.m.; « > 3 independent replicates.b:cLogD7.4 was calculated using ChemAxon MarvinSketch.c: Average fold change between 72 h IC50 values for MCF-7 and the ERa-negative cell line, values were rounded to the nearest tenth or hundredth place. Given the improvements realized with ErSO-DFP, a more complete suite of derivatives with fluorinated nitrogen heterocycles were synthesized and tested (Table 4). Note - enantiomers were separated and activity is reported for single enantiomers in this table. Derivatives (R)-22 - (R)-26 had clogD7.4 values ranging from 3.5-4.5 and amine pKavalues ranging from -2.0 to 5.4. Fluorinated nitrogen heterocycles of varying size (azetidines - 4 membered, pyrrolidines - 5 membered, piperidines - 6 membered) all exhibited anticancer activity at nanomolar potencies. In general, compounds with greater fluorination and lower pKavalues had the highest potency, with the tetrafluorinated pyrrolidine (TFPy) (R)-26, termed ErSO-TFPy, exhibiting single digit nanomolar potency (Chart 5). The opposite enantiomer, (S)-26, was also tested and found to be inactive at 1 pM. Notably, all derivatives were significantly less active when tested in ERa- cell lines (MDA-MB-231, HCT-116, HT-29). To explore the possibility of branching off these fluorinated nitrogen heterocycles, compound 27 was synthesized bearing a methyl ester at the 2-position of a difluorinated pyrrolidine, however, biological testing indicated a loss of activity and further branching was not explored.

[0130] Chart 5. Structures of R and S enantiomers of ErSO-DFP and ErSO-TFPy.

[0131] Table 4. Cell Culture Activity for Compounds 22-26.

[0132] 3To determine IC50, cancer cells were incubated with the compound for 24 or 72 h and viability was measured via alamar blue fluorescence; raptinal (100 μM) was used as the 100% dead control; data is shown as mean ± s.e.m; n = 3 independent replicates.bcLogD7.4 was calculated using ChemAxon Marvin Sketch.cAmine pK'a (i.e., tertiary amine on the final compound) was estimated using Chem Axon MarvinSketch.dAverage fold change between 72 h IC50 values for MCF-7 and ERa- negative cell line, values were then rounded to the nearest hundredth place.en ------ 2 independent replicates. Note that for compounds in this table, the absolute configuration as drawn is inferred from the known absolute configurations of ErSO and ErSO-DFP.

[0133] Linear chain derivatives. Continuing to probe SAR surrounding the B-ring, derivatives with linear chains (C-linked and N-linked) were synthesized (Table 5). C-linked alkyl chains were synthesized by forming the C-C linkage through Grignard attack on aryl ketone to give the tertiary alcohol, followed by Friedel-Crafts alkylation under acidic conditions, analogous to A-Ring derivatives. N-linked secondary amine derivatives were synthesized via the tertiary chloride, similar to the nitrogen heterocycles. Overall, 7 derivatives were synthesized with varying chain lengths and linkages (Compounds 28-34). All derivatives were tested against MCF7 cells. Surprisingly, all C- linked alkyl chains had some anticancer activity, with compounds 29 and 30 exhibiting ICso’s of 310 nM and 274 nM respectively. These compounds are less potent than the lead candidates (ErSO, ErSO- DFP, ErSO-TFPy), but their activity further demonstrates the flexibility of the B-ring to modification. Compound 28 (n-propyl derivative) was significantly less active and did not reach 50% cell death at the concentrations tested within 24 hours to generate an ICso value. Similarly to cyclic amines, all linear amines tested lacked activity (Compounds 31-34); however, unlike the cyclic amines, fluorination (Compound 34) was insufficient to rescue activity. Without knowledge of the target, it is difficult to propose rationale for these trends in activity, but given the activity of Compounds 28 & 29, it’s possible that the B-ring substituent is responsible for orienting the A-ring phenol in the active site rather than interacting directly with amino acids itself. Notably, calculated amine pKavalues were higher for linear amines (6.2-6.4) than fluorinated nitrogen heterocycle suggesting some population of charged species at physiological pH which could lead to instability of the bond between quaternary carbon and tertiary amine, which had previously been identified as a potential liability in A-linked derivatives.

[0134] Table 5. Cell culture activity for B-Ring linear chain derivatives.

[0135] Compounds were tested in MCF-7 cells for 24 hours and viability measured via alamar blue fluorescence (n > 2). cLogD7.4 and Amine pl<aof the conjugate acid calculated using ChemAxon MarvinSketch.

[0136] Importance of Fluorination. Fluorine and nitrogen heterocycles are common motifs in medicinal chemistry and drug development. The small size of fluorine makes it sterically non- disruptive and the C-F bond can have strong effects on hydrophobicity, conformation, and pKa leading to downstream effects on drug potency, permeability, and metabolism. In the context of fluorinated nitrogen heterocycles, fluorine is known to increase basicity of the nitrogen and generally favor conformations that place fluorine at the axial position and in a gauche position relative to other polar groups. In the context of 3-(4-hydroxyphenyl)indoline-2-ones, the powerful effect of fluorine has been observed in fluorinated derivatives of both the A-Ring and B-Ring. To better understand the importance of fluorination to nitrogen heterocycles in the B-Ring, compound 35 was synthesized to obtain matched fluorinated / unfluorinated derivatives of the 6-membered piperidine (16 & ErSO-DFP) and the 4- membered azetidine (35 & 25) seen in Figure 3A-B. Calculated pKas indicate a modest drop in basicity from 5.7 to 4.9 in the piperidine system and a large change from 5.0 to -0.3 in the azetidine system. Compound 16 was predicted to be 2% protonated at physiological pH (pH = 7.4) suggesting the possibility of protonation and cleavage of the C-N bond to give a neutral amine and a stabilized carbocation. Assessment of stability in phosphate-buffered saline (PBS) for 2 hours at 37 °C revealed that ErSO-DFP, Compound 35, and Compound 25 were nearly 100% stable, but only 50% of Compound 16 was remaining. Interestingly, 16 retained some activity against MCF7 cells with an Emax of -40% (1 gM at 24 hours). If 16 were 100% stable, similar to ErSO-DFP, we would expect the doseresponse curve would more closely mirror that of ErSO-DFP’ s. This study suggests that in the case of the 6-membered ring, fluorination of nitrogen heterocycles appears to increase stability by tuning amine basicity. However, in the 4-membered ring system, the complete lack of activity of Compound 35 and 100% stability in PBS indicates that fluorination may be playing a different role in the activity of Compound 25, perhaps related to permeability or target engagement. These experiments highlight the importance of amine pKa in B-Ring derivatives with a C-N bond and may explain the inactivity of Compounds 16, 19, 20, 31-34, but the importance of fluorination of B-Ring nitrogen heterocycles is still not fully understood. A more complete assessment of N-linked derivatives stability in media at longer incubations may reveal a more complete rationale for the activity of derivatives and cell-line selectivity.

[0137] Structure-Activity Relationships (Core Modifications).

[0138] Electron Withdrawing Groups. Moving forward with SAR studies of the core, the tetrafluorinated pyrrolidine of ErSO-TFPy was chosen as the optimal B-Ring motif due to the increased potency and increased cell-line selectivity. The A-Ring phenol was kept constant due to its essential nature for anticancer activity. The electronics of the ring were investigated by incorporating various electron- withdrawing groups (EWG) seen in Table 6. By decreasing the electron-richness of the indoline-2-one core, our hope was to stabilize the bond between the quaternary carbon and pyrrolidine nitrogen which, if cleaved, would generate a tertiary carbocation stabilized by resonance. Synthesis of nine derivatives (Compounds 36-44) was achieved through a similar sequence to Scheme 3, beginning with various isatin starting materials. cLogD?.4 values ranged from 3.0-5.2 and demonstrated that decreasing fluorine count on the core may be another strategy for decreasing lipophilicity. Amine pKadid not significantly shift, and new derivatives had values ranging from -1.0-0.0. Overall, 24 h ICso values in MCF-7 cells (compounds tested as racemic mixtures) varied significantly, demonstrating that the electronics of the core may be important for activity, yet all derivatives tested maintained some activity indicating that there is flexibility at this position. Fluorinated derivatives (36-38) had the greatest potency indicating that there may be a preference for fluorine or trifluoromethyl. Derivatives with the strongest electron-withdrawing character (40, 42, 44) had activity but were significantly less potent than ErSO-TFPy. Table 6. Cell culture activity for EWG-Core Derivatives.

[0139] Compounds were tested in MCF-7 cells for 24 hours and viability was measured via alamar blue fluorescence (n > 2). cL.0gD7.-i arid Amine pKaof the conjugate acid were calculated using ChemAxon Marvin Sketch.

[0140] Branching Amines. To explore the possibility of placing larger substituents on the core, we envisioned a late-stage derivatization strategy to append primary amines of varying length to the 7- position. Initial attempts to do this using Ullman-type chemistry and nucleophilic aromatic substitution (SNAT) were unsuccessful, leading to the pursuit of a Buchwald Hartwig amination strategy. Initial attempts began with the TBS-protected 7-bromo derivative (Table 7), as protic groups can interfere with cross-coupling. Generally, BrettPhos catalysts are best for primary amines, giving the mono- arylated product, thus the precatalyst BrettPhos Pd G3 was used throughout the optimization. Minimal reactivity was observed at 80 °C when using a standard base (CS2CO3) and solvent (Dioxane). Increasing reaction temperature to 120 °C only resulted in loss of the TBS group. Note - These reactions demonstrate that the bond between quaternary carbon and tertiary amine is quite temperaturestable. From here, base was switched to LiHMDS (preferred for protic groups), and couplings were attempted on the deprotected 7-bromo derivative. No reactivity was observed until solvent was switched to tetrahydrofuran (THF) at which point coupling with various amines (propylamine, butylamine, hexylamine) proceeded at 60-80 °C. Yields around 20 % were observed and no further optimization was attempted. Note - the lower yield of hexylamine is due to product loss from reversephase HPLC purification.

[0141] Table 7. Optimization of Buchwald-Hartwig Aminations.

[0142] Reactions were run for 2-16 hours and products isolated via column chromatography. TBS: tert- Butyldimethylsilyl; CS2CO3: Cesium Carbonate; LiHMDS: Lithium bis(trimethylsilyl)amide; THF: T etrahy drofuran.

[0143] This Buchwald-Hartwig strategy thus provided the 7-bromo derivative (Compound 45) as well as 3 branching-amine derivatives (46-48) as final compounds for assessment in cell culture (Table 8). Like previous derivatives, compounds were assessed as racemic mixtures in MCF7 cells. Compound 45 (7-bromo) was the most potent derivative tested with an equivalent potency to racemic ErSO-TFPy (72 h MCF-7 IC50 = 8 nM). This was surprising given the loss of activity of Compound 41 (5 -bromo, 7-bromo; 24 h MCF-7 IC50 = 340 nM) and Compound 43 (5-bromo, 7-fluoro; 24 h MCF-7 IC50 = 1100 nM). Presumably, this indicates that bromination at the 7-position is favorable while bromination at the 5-position is not. Furthermore, all branched amine derivatives (46-48) maintained potent nanomolar activity despite possessing relatively large groups at the 7-position (i.e. hexylamine). Compound 48 was less potent than Compound 46, suggesting that activity may be lost if larger groups are placed at this position, but position 7 of the core may be considered a good option appending larger groups onto the scaffold. It remains to be seen whether branching is tolerated at other positions (4, 5, & 6). Table 8. Cell culture activity for Branched Amine - Core derivatives.

[0144] Compounds were tested in MCF-7 cells for 72 hours and viability measured via alamar blue fluorescence (n > 2). cLogD7.4 and Amine pK?. of the conjugate acid calculated using ChemAxon MarvinSketch.

[0145] Comparison of ErSO, ErSO-DFP, and ErSO-TFPy in Preclinical Toxicity Assays. Preclinical toxicology studies are an important step in assessing the safety of potential therapeutics. Testing compounds in animal models (rodents, rabbits, dogs, monkeys, etc.) has historically played an important role in this process, but there is a growing understanding that these models do not always correlate with human safety and improvements are needed17. Protein microarrays, organ-on-a-chip, and artificial intelligence are being used to improve safety testing, but it is likely that multiple toxicity assays will be needed, and the best assay may be dependent on the class of therapeutics being assessed. The safety of lead molecules ErSO, ErSO-DFP, and ErSO-TFPy were assessed for hemolysis, mutagenicity, and in 2-D cell culture against “normal” cell lines.

[0146] Hemolysis Assay. Assessment of compound toxicity to erythrocytes (red blood cells) using a hemolysis assay is a simple and initial step in toxicity assessment of many small molecule therapeutics. Compounds that interfere with the cell membrane of red blood cells will release hemoglobin which has a distinct absorbance that can easily be measured with standard plate readers. Greater surfactant / detergent character of small molecules leads to an increased likelihood of hemolysis and poor translation to humans. Toxicity of ErSO, ErSO-DFP, and ErSO-TFPy to red blood cells was assessed at increasing concentrations (12.5-400 pM), and DMSO and Triton-X were used as negative and positive controls respectively (Figure 4). ErSO demonstrated minimal hemolysis at lower micromolar concentrations but did induce significant hemolysis when tested from 50-400 pM. The hemolysis profile was improved for ErSO-DFP (minimal hemolysis up to 200 pM) and ErSO-TFPy (minimal hemolysis up to 100 pM). Ultimately, all three molecules exhibited minimal hemolysis at 12.5 pM, well above the nanomolar concentrations required for anticancer activity and do not have the detergent-like character associated with most hemolytic agents.

[0147] Ames Testing. Assessment of the mutagenicity of new drugs is required by regulatory authorities to screen for potential carcinogenic activity. The Ames mutagenicity test is the standard assay for assessing mutagenicity of new compounds. Briefly, S. typhimurium and E. coli mutants are used that are auxotrophic for histidine and tryptophan, respectively. When dosed with mutagenic compounds, mutations in bacterial DNA can occur such that individual auxotrophic bacteria can be reverted to prototrophs capable of uncontrolled proliferation. Replicating bacteria will cause a change in pH, resulting in a color change that is considered indicative of mutagenicity. ErSO, ErSO-DFP, and ErSO-TFPy were first tested in S. typhimurium strain 14028 and E. coli strain MG1655 to ensure bacterial survival throughout the test, and the MICs were > 250 pM. Mutagenicity of the compounds was assessed in three strains of S. typhimurium'. TA100 (positive control = 5 pg / mL NaNs), TA98 (positive control = 300 pg / mL 2-nitrofluorene) and TA97a (positive control = 0.8 pg / mL 9- aminoacridine hydrochloride). 50 pM ErSO induced a similar number of revertant colonies as the background in TA100 / TA98 and less than the background in TA97a, suggesting possible antibacterial activity (Example 3, Table 16). Assessment of ErSO-DFP found a significantly increased number of revertant colonies in the TAI 00 strain and background levels in TA98 and TA97a. ErSO-TFPy exhibited background levels of reversion in all three strains. Overall, ErSO-DFP exhibited the greatest level of mutagenicity at 50 pM, but only in one strain and at much higher concentrations than those required for anticancer activity.

[0148] Evaluation in “Normal” Cell Lines. Evaluation of small molecules in 2-D cell culture is standard practice for initial assessments of anticancer activity and toxicity. Cell cultures may be primary, isolated and passaged directly from patients, or established / immortalized cell lines that proliferate indefinitely due to chromosomal aberrations that have occurred naturally or introduced artificially (HPV E6 / E7, hTERT, etc.). Primary cell lines are considered more physiologically relevant and recapitulate tumor heterogeneity better than immortalized cell lines, but immortalized cell lines are used more frequently due to their unlimited replicative potential, ease of passaging, and reproducibility between laboratories. Immortalized cell lines may be established from tumors (cancer cells) or healthy tissues (normal cells). Evaluation of therapeutics against “normal” cell lines is a common initial assessment of toxicity, although it is important to note that this may be more appropriate for antibiotics than cancer therapeutics due to similarities in biology between proliferating cancerous and healthy cells. ErSO, ErSO-DFP, and ErSO-TFPy were all evaluated for activity against “normal” cell lines using immortalized fibroblast cell lines HFF-1 (foreskin fibroblasts), IMR90 (lung fibroblasts), Hs68 (foreskin fibroblasts) and primary human T-cells (Figure 5A-C, Table 9). ErSO and ErSO-TFPy demonstrated nanomolar activity against HFF-1, IMR90, and Hs68 fibroblasts, but were significantly less active against primary T cells. ErSO-DFP had activity against HFF-1 cells but was significantly less potent in IMR90 and Hs68 cells and was similarly inactive against primary T-cells. These results are considered promising given the lack of activity against primary cells and selectivity of ErSO-DFP for breast cancer cells (MCF-7) vs normal fibroblasts (HFF-1, IMR90, and Hs68).

[0149] Table 9. Calculated ICsos for each compound / cell line.

[0150] Primary T-cells isolated from 30-year-old African American male donor.

[0151] Acid Stability. The majority of marketed drugs are orally bioavailable, and this is generally the preferred route of administration due to patient compliance, dosing flexibility, and convenience. ErSO- DFP has poor oral bioavailability (relative to ErSO), and this is thought to be due to instability in the acidic environment of the gut, which may promote cleavage of the bond between quaternary carbon and tertiary nitrogen. The ideal compound would have the oral bioavailability of ErSO combined with the selectivity and tolerability of ErSO-DFP and ErSO-TFPy. Simulated gastric fluid (SGF) supplemented with pepsin recapitulates the acidic environment of the gut, and assessing stability of derivatives in SGF can provide insight on which derivatives may be orally bioavailable (Figure 6). ErSO, ErSO-TFPy and compounds 36-48 were assessed for SGF stability over 2 hours using LCMS to quantify remaining compound. ErSO was stable with 89 % of compound remaining after 2 hours while ErSO-TFPy was unstable (19 % remaining), similar to previously reported values. Compounds 36-48 demonstrated a broad range of stability with % remaining ranging from 6-93 %, supporting the hypothesis that tuning the electronics of the core could modulate acid stability. Unfortunately, the compounds of greatest interest that had demonstrated potency similar to ErSO / ErSO-TFPy (Compounds 36, 37, 45) were the least stable (% remaining = 6-7 %). Compound 40 (5-nitro, 7-fluoro) demonstrated the greatest increase in acid stability (93 %, comparable to ErSO) and has an ICso of 240 nM when assessed in MCF-7 cells. Given the potency of oxyphenisatin acetate, ICso ~ 600 nM in MCF- 7 cells, this activity may be sufficient given a favorable pharmacokinetic profile. Future studies are needed to confirm oral bioavailability and efficacy of 40 in an oral-dosing model for breast cancer.

[0152] Chart 6. Stability of ErSO, ErSO-TFPy, and core derivatives in simulated gastric fluid (SGF) supplemented with pepsin (Figure 6). Alternative Derivatives.

[0153] Decreasing Fluorination. One of the interesting findings of the medicinal chemistry campaigns performed on this scaffold is the powerful effect of fluorine. Strategic fluorination / trifluoromethylation of BHPI led to a significant boost in potency and the discovery of ErSO. Fluorination of B-Ring nitrogen heterocycles lead to the discovery of ErSO-DFP and ErSO-TFPy. As mentioned previously, fluorine has numerous effects and can improve compound potency, permeability, and stability. While the C-F bond is considered stable, in some cases intramolecular and biological nucleophiles may displace a fluorine; undesired fluoride release can cause issues such as skeletal fluorosis. As ErSO, ErSO-DFP, and ErSO-TFPy have relatively high fluorine counts, we have been interested in alternative derivatives that contain fewer fluorine atoms. One such compound is the 7-bromo derivative, Compound 45 which had an IC50 of 8 nM in MCF-7 when tested as a racemic mixture. To further investigate this compound, enantiomers were separated via chiral chromatography to afford (R)-45 and (S)-45 (Table 10). (R)-45 has equivalent potency to ErSO-TFPy (MCF7 ICso = 3 nM) and (S)-45 is inactive as expected. Both enantiomers have high ICso values in MDA-MB-231 cells (25-30 pM), and this activity is expected to be an alternative mechanism of cell death. Preliminary tolerability experiments suggested that (7?)-45 is well tolerated in mice (MTDiv > 50 mg / kg).

[0154] Table 10. Chiral Separation of Compound 45 and Evaluation of Cell Culture Activity.

[0155] Compounds were tested in MCF-7 / MDA-MB-231 for 72 hours and viability measured via alamar blue fluorescence (n > 2). Mouse tolerability assessed in female CD-I mice via tail vein injection (n=3). Formulation: 2.5% Ethanol, 5% Kolliphor EL, 15% Propylene Glycol, 77.5% Sterile saline MTDiv: Maximum Tolerated Dose (intravenous administration).

[0156] Prodrug Strategy. Prodrugs are compounds that possess little biological activity themselves, yet they are able to be transformed in the body to the active species upon cleavage of group(s) that mask essential components of the molecule. This strategy is commonly used in drug development for various reasons including increasing solubility, improving stability, modulating metabolism, decreasing toxicity, and selective organ delivery. As mentioned previously, oxyphenisatin is often tested as the prodrug oxyphenisatin acetate. The acetylated phenols are expected to be hydrolyzed or cleaved by esterases in cells to liberate oxyphenisatin. Although it is not clear what benefits are realized with oxyphenisatin acetate, we were interested in applying this strategy to ErSO-TFPy, believing it may provide a similar anticancer profile with the potential to improve pharmacokinetics and tolerability in future experiments. Compound 26 was acetylated with acetic anhydride to provide 49, which was separated via chiral chromatography (Table 11). Assessment in cell culture revealed (R)-49 exhibited equivalent IC50 s to ErSO-TFPy in MCF-7 cells at 24 hours and 72 hours, indicating that the acetyl group is cleaved relatively quickly. As expected, both enantiomers require much higher concentrations to kill MDA-MB-231 cells. Preliminary tolerability experiments suggested that (R)-49 is well tolerated in mice (MTDiv > 50 mg / kg).

[0157] Table 11. Chiral Separation of Compound 49 and Evaluation of Cell Culture Activity and Mouse Tolerability.

[0158] Compounds were tested in MCF-7 / MDA-MB-231 for 72 hours and viability measured via alamar blue fluorescence (n > 2). Mouse tolerability assessed in female CD-I mice via tail vein injection (n=3). Formulation: 2.5% Ethanol, 5% Kolliphor EL, 15% Propylene Glycol, 77.5% Sterile saline MTDiv: Maximum Tolerated Dose (intravenous administration).

[0159] Conclusions. Structure-activity relationship studies are important for identifying the pharmacophore of small molecule scaffolds and informing medicinal chemistry strategies for drug optimization. Our SAR studies on the 3-(4-hydroxyphenyl)indoline-2-one scaffold have confirmed the potent and selective anticancer activity, differential activity between enantiomers, and the importance of the phenol while revealing new trends like the power of lipophilicity, the tunable electronics of the core, and the diversity of chemical groups tolerated at the B-Ring. Efforts to modify the A-Ring phenol were largely unsuccessful with even small changes (fluorination, methylation, etc.) resulting in dramatic loss of activity. The flexibility of the B-Ring allowed us to incorporate fluorinated nitrogen heterocycles to decrease lipophilicity leading to the discovery of more selective derivatives ErSO-DFP and ErSO- TFPy. The role of fluorination of nitrogen heterocycles is still unclear but appears to play a role in the stability of the bond between the quaternary carbon and tertiary amine. Studies on the core revealed a wide range of activities depending on the EWG identity and placement and identified the 7-position as a potential place for branched groups. ErSO-DFP appears to be more selective for breast cancer cell lines compared to “normal” cell lines, while ErSO-TFPy exhibits very potent anticancer activity, killing cells at single digit nanomolar concentrations. Newer derivatives (R)-45 and (R)-49 may be good alternatives to ErSO-TFPy for decreasing fluorine count and altering pharmacokinetics respectively, but the primary issue with ErSO-DFP and ErSO-TFPy identified in these experiments appears to be acid instability / poor oral bioavailability, thus future medicinal chemistry efforts should focus on improving these attributes. An alternative strategy to improve acid stability by linking nitrogen heterocycles to scaffold through a C-C linkage was also investigated (Table 12). Initial derivatives 51- 54 were stable but lacked anticancer activity. The boc-protected amines (51, 53) are likely inactive due to steric bulk while the free secondary amines (52, 54) may be too polar or reduce lipophilicity to a greater extent than optimal. Future derivatives could improve on this by methylating or trifluoromethylating the amine to tune polarity / lipophilicity.

[0160] Table 12. Cell culture activity for additional derivatives (27, 51-54).

[0161] Compounds were tested in MCF-7 cells for 24 hours and viability measured via alamar blue fluorescence (n > 2). cLogDv.4 and Amine pKa of the conjugate acid calculated using ChemAxon Marvin Sketch.

[0162] Noncovalent Probe Synthesis and Evaluation. Drugs are generally assumed to covalently modify target proteins if they possess electrophilic warheads (epoxides, a,p-unsaturated carbonyls, sulfones) that can react with nucleophilic species (e.g. cysteine). ErSO molecules do not contain an obvious electrophilic functional group. Thus, it is assumed to utilize a noncovalent mechanism of action. In designing initial noncovalent probe molecules, modifications were focused primarily on highly manipulatable B-Ring. Probe molecules 55-57 were synthesized bearing an alkyne (suitable for click chemistry) or biotin (suitable for biotin-streptavidm pulldowns) and assessed for activity in cell culture (Table 13). Probe 55 maintained cellular potency (24 h MCF7 ICso = 56 nM; note this compound is racemic so the active single enantiomer is likely ~28 nM), while the sterically larger modifications of probes 56 and 57 were devoid of activity. The loss of activity of probe 56 (differs from 55 by two methylene groups) highlights the restrictive SAR for these compounds; an early indication of future challenges in probe syntheses, biological evaluations, and target identification.

[0163] Table 13. Cell culture activity for noncovalent probes.

[0164]

[0165] Compounds / probes were tested in MCF7 cells for 24 hours and viability measured via alamar blue fluorescence (n > 2).

[0166] Following difficulties with chiral separation of probe 55, enantiomers were obtained by coupling the phenol to boc-glycine, chiral separation, and saponification to afford (R)-55 and (S)-55. Cell culture activity was used to establish stereochemistry and (R)-55 had comparable MCF7 potency to ErSO while (S)-55 was inactive (Table 14). Both enantiomers were inactive when tested against MDA-MB-231 as expected. The loss of activity of 56 indicated that immobilization of the probe through the alkyne would likely disrupt engagement with the biological target. For this reason, we adapted a protocol normally reserved for covalent probes -- incubating cells / lysate with probe followed by click chemistry with biotin-azide and enrichment using streptavidin beads. Unfortunately, no protein was observed in elution fractions decreasing confidence in this this modified approach. Copper is cytotoxic to cells and may denature proteins depending on concentration which may also contribute to the failure of this noncovalent approach.

[0167] Table 14. Cell culture activity for enantiomers of alkynylated probe 55.

[0168] Compounds / probes were tested in MCF7 / MDA-MB-231 cells for 24 hours and viability measured via alamar blue fluorescence (n > 2). Note - Stereochemistry is inferred from biological activity.

[0169] Photoaffinity Probe Synthesis and Evaluation. We next focused on the design and synthesis of photoaffinity probes, which are modified with an enrichment tag like alkyne / biotin as well as a photoaffinity group like aryl azides / diazrines (Table 15). As mentioned, under UV-irradiation these groups will form a covalent linkage with nearby amino acid residues. This now covalent bond between compound and target protein is stable to harsher conditions (e.g., copper click chemistry and protein denaturation). Benzophenone-based probes were not pursued due to their large steric bulk and the limiting SAR of the scaffold. A terminal alkyne at the 7-position of the core was tolerated (derivative not shown), but all diazirine-based probes (58-61) were inactive in MCF7 cells. However, several aryl azide probes (62-64) maintained potent antiproliferative activity. Probe 62 was not pursued as it was found to be unstable at room temperature. This instability is likely resulting from an intramolecular cyclization between aryl azide and propargyl ether, a known spontaneous chemical reaction. However simply removing the ether linkage and replacing it with an alkyne, yielded probe 63 which was stable. Probe 65, which possessed a large biotin moiety at the meta-position of the B-ring was not active. Initial fluorescent labelling efforts focused on probe 63 and enantiomers (R)-63 and (N)-63 were isolated via preparative chiral chromatography.

[0170] Table 15. Cell culture activity for photoaffinity probes.

[0171]

[0172] Compounds / probes were tested in MCF7 cells for 24-72 hours and viability measured via alamar blue fluorescence (n > 2). PA - photoaffinity.

[0173] Diazirine Probe. While thermal shift experiments support binding between ErSO molecules and TRPM4, the fact that these experiments are done in live cells mean that it remains possible that cellular effects elicited by ErSO molecules could be responsible for destabilization of TRPM4. One such possibility would be that ErSO molecules bind a complex involving TRPM4 leading to a conformational change and TRPM4 destabilization. Furthermore, multiple pieces of evidence of binding should be provided to validate a biological target. For this reason, we refocused efforts on obtaining an active diazirine probe. One of the important synthetic advancements was finding that the tertiary alcohol may be reduced to the tertiary hydride, which can be deprotonated to bring electrophiles in under basic conditions (Scheme 4). Previously discussed, C-linked alkyl chains are tolerated at the B-Ring and this new chemistry allowed for nucleophilic attack on 3-(But-3-yn-l-yl)-3-(2-iodoethyl)- 3H-diazirine, which was commercially available. Deprotection afforded the racemic mixture (probe 66), which could be separated by chiral chromatography to provide (S)-66 and (R)-66. Testing in MCF7 confirmed nanomolar potency / enantiomer specificity, providing the first active diazirine probe (Scheme 4). With this probe in hand, biotin-streptavidin / proteomic experiments will be redone with a higher likelihood of success than previous aryl azide-based probes. Scheme 4. Synthesis and cell culture activity of diazirine probe 64.

[0174] (A) wBuLi - n-Butyllithium. THF - Tetrahydrofuran. SnCl2 - Tin II Chloride. AcOH - Dimethylformamide. KO / Bu - Potassium tert-butoxide. KHF2 - Potassium hydrogen difluoride. MeOH - Methanol. RT - Room temperature. (B) Enantiomers separated via chiral chromatography and evaluated in cell culture. MCF7 cells treated with compound and cell viability measured via alamar blue fluorescence at 72 h. (n > 2).

[0175] Pharmaceutical Formulations.

[0176] The compounds described herein can be used to prepare therapeutic pharmaceutical compositions, for example, by combining the compounds with a pharmaceutically acceptable diluent, excipient, or carrier. The compounds may be added to a carrier in the form of a salt or solvate. For example, in cases where compounds are sufficiently basic or acidic to form stable nontoxic acid or base salts, administration of the compounds as salts may be appropriate. Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids that form a physiologically acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, a-ketoglutarate, and β-glycerophosphate. Suitable inorganic salts may also be formed, including hydrochloride, halide, sulfate, nitrate, bicarbonate, and carbonate salts.

[0177] Pharmaceutically acceptable salts may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid to provide a physiologically acceptable ionic compound. Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example, calcium) salts of carboxylic acids can also be prepared by analogous methods.

[0178] The compounds of the formulas described herein can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient, in a variety of forms. The forms can be specifically adapted to a chosen route of administration, e.g., oral or parenteral administration, by intravenous, intramuscular, topical or subcutaneous routes.

[0179] The compounds described herein may be systemically administered in combination with a pharmaceutically acceptable vehicle, such as an inert diluent or an assimilable edible carrier. For oral administration, compounds can be enclosed in hard or soft shell gelatin capsules, compressed into tablets, or incorporated directly into the food of a patient's diet. Compounds may also be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations typically contain at least 0.1% of active compound. The percentage of the compositions and preparations can vary and may conveniently be from about 0.5% to about 60%, about 1% to about 25%, or about 2% to about 10%, of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions can be such that an effective dosage level can be obtained.

[0180] The tablets, troches, pills, capsules, and the like may also contain one or more of the following: binders such as gum tragacanth, acacia, com starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as com starch, potato starch, alginic acid and the like; and a lubricant such as magnesium stearate. A sweetening agent such as sucrose, fructose, lactose or aspartame; or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring, may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propyl parabens as preservatives, a dye and flavoring such as cherry or orange flavor. Any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release preparations and devices.

[0181] The active compound may be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can be prepared in glycerol, liquid polyethylene glycols, triacetin, or mixtures thereof, or in a pharmaceutically acceptable oil. Under ordinary conditions of storage and use, preparations may contain a preservative to prevent the growth of microorganisms.

[0182] Pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions, dispersions, or sterile powders comprising the active ingredient adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. The ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions, or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and / or antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers, or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by agents delaying absorption, for example, aluminum monostearate and / or gelatin.

[0183] Sterile injectable solutions can be prepared by incorporating the active compound in the required amount in the appropriate solvent with various other ingredients enumerated above, as required, optionally followed by fdter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation can include vacuum drying and freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the solution.

[0184] For topical administration, compounds may be applied in pure form, e.g., when they are liquids. However, it will generally be desirable to administer the active agent to the skin as a composition or formulation, for example, in combination with a dermatologically acceptable carrier, which may be a solid, a liquid, a gel, or the like.

[0185] Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina, and the like. Useful liquid carriers include water, dimethyl sulfoxide (DMSO), alcohols, glycols, or water-alcohol / glycol blends, in which a compound can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using a pump-type or aerosol sprayer.

[0186] Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses, or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user.

[0187] Examples of dermatological compositions for delivering active agents to the skin are known to the art; for example, see U.S. Patent Nos. 4,992,478 (Geria), 4,820,508 (Wortzman), 4,608,392 (Jacquet et al.), and 4,559,157 (Smith et al.). Such dermatological compositions can be used in combinations with the compounds described herein where an ingredient of such compositions can optionally be replaced by a compound described herein, or a compound described herein can be added to the composition.

[0188] Useful dosages of the compounds described herein can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Patent No. 4,938,949 (Borch et al.). The amount of a compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular compound or salt selected but also with the route of administration, the nature of the condition being treated, and the age and condition of the patient, and will be ultimately at the discretion of an attendant physician or clinician.

[0189] In general, however, a suitable dose will be in the range of from about 0.5 to about 100 mg / kg, e.g., from about 10 to about 75 mg / kg of body weight per day, such as 3 to about 50 mg per kilogram body weight of the recipient per day, preferably in the range of 6 to 90 mg / kg / day, most preferably in the range of 15 to 60 mg / kg / day.

[0190] The compound is conveniently formulated in unit dosage form; for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, most conveniently, 50 to 500 mg of active ingredient per unit dosage form. In one embodiment, the invention provides a composition comprising a compound of the invention formulated in such a unit dosage form.

[0191] The compound can be conveniently administered in a unit dosage form, for example, containing 5 to 1000 mg / m2, conveniently 10 to 750 mg / m2, most conveniently, 50 to 500 mg / m2of active ingredient per unit dosage form. The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more subdoses per day. The sub-dose itself may be further divided, e.g., into a number of discrete, loosely spaced administrations.

[0192] The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye.

[0193] The invention provides methods of treating cancer in a mammal, which involve administering to a mammal having cancer an effective amount of a compound or composition described herein. A mammal includes a primate, human, rodent, canine, feline, bovine, ovine, equine, swine, caprine, bovine and the like. Cancer refers to various types of malignant neoplasm, for example, colon cancer, breast cancer, melanoma and leukemia, and in general is characterized by an undesirable cellular proliferation, e.g., unregulated growth, lack of differentiation, local tissue invasion, and metastasis. The following Examples are intended to illustrate the above invention and should not be construed as to narrow its scope. One skilled in the art will readily recognize that the Examples suggest many other ways in which the invention could be practiced. It should be understood that numerous variations and modifications may be made while remaining within the scope of the invention.

[0194] EXAMPLES

[0195] Example 1. Materials and Methods.

[0196] Cell Lines and Culturing Conditions. All cell lines cultured at 37 ' C with 5% CO2. All cells were grown in medium lacking phenol-red. MCF-7, IMR90 cells were grown in Eagles Minimum Essential Media (EMEM) supplemented with 10% Fetal Bovine Serum (FBS) and 1% Penicillin- Streptomycin (P / S). MDA-MB-231 cells were grown in RPMI 1640 supplemented with 10% FBS and 1% P / S. HFF1, Hs68 cells were grown in Dulbecco’s Minimum Essential Media (DMEM) supplemented with 10% FBS and 1% P / S. Primary T cells isolated from 30 year old African American male using STEMCELL Human T-cell Isolation Kit and cultured using ImmunoCult-XF T cell expansion media supplemented with 10 ng / mL hIL2. All other cell lines were used directly from ATCC stocks and / or submitted to University of Arizona Genetics Core (UAGC) orlDEXX for authentication via STR profiling.

[0197] Alamar Blue Fluorescence for Cell Viability (IC50). 6,000-10,000 cells were seeded per well in 99 pL of appropriate media in 96-well plates and allowed to adhere overnight. 1 pL of compoundcontaining DMSO solution was added to each well to give a final volume of 100 pL (DMSO final concentration = 1%). Compounds were incubated for 24 hours- 168 hours before aspirating media and replacing with fresh media (100 pL). 10 pL of Alamar blue solution (1 mg resazurin dissolved in 10 mL of PBS) was added to each well. After 4-6 hours of incubation, fluorescence ( / .excitation = 555 nm, / emission = 585 nm) was measured using a SpectraMax M3 plate reader (Molecular Devices). 5 technical replicates per concentration. Percent dead was calculated using 100 pM Raptinal as a 100% dead control. Dose response curves and IC50 values were calculated using Origin Pro V10.

[0198] PBS Stability. PBS (pH 7.4) was warmed to 37 °C and 2970 pL was then aliquoted into multiple Eppendorf tubes followed by 30 pL of 10 mM DMSO stock solutions of test compounds. Samples were incubated at 37 °C for the entire experiment with vortexing every 15 min. At each desired time point (0 and 120 min), a 500 pL aliquot is removed and quenched into 250 pL of acetonitrile. Samples are stored at 4 °C until all time points are complete. Samples were filtered and concentrations were determined by LC-MS. BHPI (100 pM or 10 pM) was added to each sample immediately before injection as an internal standard and absorbances at 254 nm measured. Percent remaining was calculated by comparing peak integrations at time 0 and time 2 hours.

[0199] Hemolysis Assay. 100 pL of whole blood was combined with 500 pL of saline and centrifuged for 5 minutes at 4 °C at 300x g. The supernatant was aspirated and cells were resuspended in 500 pL saline and the process repeated 3 times to wash cells. Cells were then resuspended in 800 pL red blood cell buffer (10 mM NaiHPO-i, 150 mM NaCl, 1 mM MgCh, pH 7.4). 19 pL of red blood cell buffer was added to PCR tubes followed by 1 pL of DMSO stock solution (compound). 30% Triton-x-100 in water was used as positive control. The tubes were vortexed to mix. Then, 10 pL of washed red blood cells were added to each tube. Tubes were incubated at 37 °C for 2 hours and then centrifuged for 5 minutes at 300x g (4 °C). 20 pL of the supernatant was transferred to well of a clear bottom 384 well plate and absorbance read at X = 540 nM. Th percent lysis was normalized to the positive control at 100%. All samples were done in triplicate.

[0200] Ames Mutagenicity. Ames testing was performed at the indicated concentrations in S. typhimurium strains using Muta-ChromePlate Kit Version 2.2 according to manufacturer’s instructions. Briefly, bacteria were plated in 96-well plates, treated with compound and indicator, followed by incubation for 4-5 days. Revertant colonies identified colorimetrically.

[0201] Simulated Gastric Fluid (SGF) Assay. SGF was prepared fresh by adding .8 g of pepsin (Sigma- Aldrich P7000) to 250 mL of SGF (Fischer Scientific 7108-16), and the pH was adjusted to 1.2 using a pH meter. This SGFwith pepsin solution was then warmed to 37 °C. Warmed SGFwith pepsin (2970 pL) was then aliquoted into multiple Eppendorf tubes followed by 30 pL of 10 mM DMSO stock solutions of test compounds. Samples were incubated at 37 °C for the entire experiment with vortexing every 15 min. At each desired time point (0 and 120 min), a 500 pL aliquot is removed and quenched into 250 pL of acetonitrile. Samples are stored at 4 °C until all time points are complete. Samples were then vortexed and then clarified at 3000 RPM for 10 min in a 4 °C centrifuge. Supernatants were collected and compound concentrations were determined by LC-MS. BHPI (100 pM or 10 pM) was added to each sample immediately before injection as an internal standard and absorbances at 254 nm measured. Percent remaining was calculated by comparing peak integrations.

[0202] Tolerability Experiments. For mouse tolerability, (R)-45 and ( / ?)-49 were formulated in in 2.5% Ethanol, 5% KolliphorEL, 15% Propylene Glycol, 77.5% Sterile saline and administered intravenously to female CD-I mice via tail-vein injection (n=3). Rodents were monitored for signs of weight loss, distress, lethargy, and neurotoxicity.

[0203] Example 2. General Chemical Synthetic Methods.

[0204] All reactions described herein were conducted under inert atmosphere (argon gas) unless otherwise noted. Chemical reagents were purchased from commercial sources and used without further purification. All solvents used were anhydrous and obtained from an SDS system or purchased from commercial sources. Compounds tested in vitro were dried overnight via lyophilization from acetonitrile.1H NMR,13C NMR,19F NMR experiments were conducted on a Bruker Advance III HD 500 MHz NMR with a CryoProbe. Spectra obtained in CD3OD were referenced for 3.31 ppm and 49.00 ppm for 'H and13C NMR spectra respectively. Spectra obtained in CDCh were referenced for 7.26 ppm and 77.16 ppm for 1H and 13C NMR spectra respectively. All NMR chemical shifts are reported in ppm (8), coupling constants (J, Hz), and peaks reported as: s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet. High resolution mass spectra (HRMS) were obtained at the UIUC SCS Mass Spectrometry Laboratory utilizing electrospray ionization (ESI). Enantiomers were separated using a Teledyne ISCO ACCQPrep HP125 Preparative HPLC (Column: Lux® 5 pM Cellulose-1, LC Column,

[0205] 250 x 21.2 mm, AXIA™ Packed, isocratic elution isopropanol / hexanes OR CHIRALPAK ADH, SFC, 5 pM, 250 x 21.2 mm, isocratic elution isopropanol / hexanes).

[0206] Scheme 5. Synthesis of Compounds 27, 31-35.

[0207] Scheme 6. Synthesis of Compounds 28-30.

[0208] Scheme 7. Synthesis of Compounds 36-44. Scheme 8. Synthesis of Compounds 45-48.

[0209] Scheme 9. Synthesis of Compounds 51-54.

[0210] Methyl (2S)-4,4-difluoro-l-(3-(4-hydroxyphenyl)-2-oxo-7-(trifluoromethyl)indolin-3- yl)pyrrolidine-2-carboxylate (27 ORMPM 03 187). In a flame dried flask under argon, 14 (211 mg, .5 mmol, 1 eq.) was dissolved in tetrahydrofuran (5 mL) and cooled to 0° C. Pyridine (120 uL, 1.5 mmol, 3 eq.) was added dropwise followed by thionyl chloride (182 uL, 2.5 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0211] In a separate flame dried flask, L-Proline, 4,4-difluoro-, methyl ester, hydrochloride salt (100 mg, .5 mmol, 1 eq.) and cesium carbonate (456 mg, 1.4 mmol, 2.8 eq.) were stirred in dichloromethane (5 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (5 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred for 48 hours. Reaction was quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in tetrahydrofuran (2 mL) under argon and 1 M tetrabutylammonium fluoride (3 eq.) was added dropwise. Reaction stirred for 1 hours at room temperature and quenched with sodium bicarbonate, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Characterized and tested as mixture of diastereomers. Isolated yield: 25 milligrams, 11% yield over 3 steps.

[0212] 'H NMR (CD3OD, 500 MHz) 8: 7.64 (d, J = 7.5 Hz, 1H), 7.56 - 7.45 (m, 3H), 7.42 - 7.35 (m, 4H), 7.14 (dt, J = 16.2, 7.8 Hz, 2H), 6.76 (dd, J = 14.9, 8.8 Hz, 3H), 4.24 (dd, J = 9.5, 3.4 Hz, 1H), 3.76 - 3.53 (m, 2H), 3.51 (s, 3H), 3.38 (q, J= 11.9 Hz, 1H), 3.23 (s, 3H), 3.08 - 2.97 (m, 1H), 2.81 - 2.55 (m, 2H), 2.36 (tt, J= 18.4, 14.4 Hz, 2H).

[0213] 19F NMR (CD3OD, 471 MHz) 8: -63.03 (d, J= 11.2 Hz), -98.74 (dq, J = 27 A, 13.8 Hz). HRMS (ESI): m / z calc, for C21 H17 N2 04 F5 Na [M+H]+ 479.1006, found: 479.1003.

[0214] 3-hydroxy-3-propyl-(trifluoromethyl)indolin-2-one (28a OR MPM 06 39). In an oven-dried flask 7-trifluoromethyl isatin (300 mg, 1.4 mmol, 1 eq.) was dissolved in tetrahydrofuran (10 mL) and cooled to -78° C. A IM solution of n-propylmagnesium bromide in ether (4.18 mL, 4.18 mmol, 3 eq.) was added drop-wise and reaction was stirred for 1 hour at -78° C. Reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 93 milligrams, 26% yield.

[0215] 'H NMR (CD3OD, 500 MHz) 8: 7.55 (d, J= 7.4 Hz, 1H), 7.50 (d, J= 8.0 Hz, 1H), 7.19 (t, J= 7.7 Hz, 1H), 2.05 - 1.80 (m, 2H), 1.27 - 1.09 (m, 1H), 1.07 - 0.93 (m, 1H), 0.85 (t, J= 7.4 Hz, 3H).

[0216] 13C NMR (CD3OD, 126 MHz) 8: 182.07, 140.39 (q, J = 2.2 Hz), 135.02, 128.69, 126.74 (q, J = 4.5 Hz), 125.14 (q, J= 270.8 Hz), 123.50, 113.38 (q, J= 33.2 Hz), 76.56, 41.12, 17.58, 14.42.

[0217] 19F NMR (CD3OD, 471 MHz) 8: -62.94.

[0218] HRMS (ESI): m / z calc, for C12 Hl 1 N 02 F3 [M-H]- 258.0742, found: 258.0746.

[0219] 3-(4-hydroxyphenyl)-3-propyl-7-(trifluoromethyl)indolin-2-one (28 ORMPM 03 269). In an oven-dried flask, 28a (152 mg, .58 mmol, 1 eq.) was dissolved in dichloroethane (3 mL). Phenol was added (272 mg, 2.9 mmol, 5 eq.) followed by p-toluenesulfonic acid (749 mg, 4.35 mmol, 7.5 eq.). Reaction was heated at 90° C for 2-4 hours and then cooled to room temperature. Solid was filtered off and crude mixture was purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 101 milligrams, 52% yield.

[0220] 'H NMR (CD3OD, 500 MHz) 8: 7.50 (d, J= 8.1 Hz, 1H), 7.38 (d, J= 7.5 Hz, 1H), 7.20 (t, J= 7.8 Hz, 1H), 7.10 (d, J = 8.9 Hz, 2H), 6.72 (d, J= 8.9 Hz, 2H), 2.38 - 2.25 (m, 1H), 2.22 - 2.15 (m, 1H), 1.26 - 1.08 (m, 1H), 0.88 (m, 4H).

[0221] 13C NMR (CD3OD, 126 MHz) 8: 183.36, 157.93, 140.57 (d, J = 2.2 Hz), 137.37, 132.04, 129.46, 128.79, 125.57 (q, J = 4.5 Hz), 125.27 (q, J = 275.1 Hz), 123.35, 116.39, 113.35 (q, J = 33.2 Hz), 57.17, 40.31, 18.89, 14.45.

[0222] 19F NMR (CD3OD, 471 MHz) 8: -62.94.

[0223] HRMS (ESI): m / z calc, for C18 H15 N 02 F3 [M-H]- 334.1055, found: 334.1053.

[0224] 3-butyl-3-hydroxy-7-(trifluoromethyl)indolin-2-one (29a OR MPM 06 41). In an oven-dried flask 7-trifluoromethyl isatin (300 mg, 1.4 mmol, 1 eq.) was dissolved in tetrahydrofuran (10 mL) and cooled to -78° C. A IM solution of n-butylmagnesium bromide in tetrahydrofuran (4.18 mL, 4.18 mmol, 3 eq.) was added drop-wise and reaction was stirred for 1 hour at -78° C. Reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 106 milligrams, 28% yield.

[0225] 'H NMR (CD3OD, 500 MHz) 8: 7.55 (d, J= 7.4 Hz, 1H), 7.50 (d, J= 8.1 Hz, 1H), 7.20 (t, J= 7.7 Hz, 1H), 1.94 (m, 2H), 1.27 (m, 2H), 1.18 - 1.05 (m, 1H), 0.94 (m, 1H), 0.83 (t, J= 7.4 Hz, 3H).

[0226] 13C NMR (CD3OD, 126 MHz) 8: 182.06, 140.41 (q, J = 2.3 Hz), 135.02, 128.68, 126.76 (q, J = 4.5 Hz), 125.15 (q, J = 270.9 Hz), 123.51, 113.39 (q, J= 33.2 Hz), 76.57, 38.62, 26.36, 23.76, 14.13.

[0227] 19F NMR (CD3OD, 471 MHz) 8: -62.95.

[0228] HRMS (ESI): m / z calc, for C13 H13 N 02 F3 [M-H]- 272.0898, found: 272.0909.

[0229] 3-butyl-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2-one (29 OR MPM 03 271). In an oven-dried flask, 29a (181 mg, .66 mmol, 1 eq.) was dissolved in dichloroethane (3 mL). Phenol was added (309 mg, 3.3 mmol, 5 eq.) followed by p-toluenesulfonic acid (852 mg, 4.9 mmol, 7.5 eq.). Reaction was heated at 90° C for 2-4 hours and then cooled to room temperature. Solid was filtered off and crude mixture was purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 100 milligrams, 44% yield.

[0230] 'H NMR (CD3OD, 500 MHz) 8: 7.50 (d, J= 8.1 Hz, 1H), 7.38 (d, J = 7.4 Hz, 1H), 7.19 (t, J = 7.8 Hz, 1H), 7.10 (d, J = 8.8 Hz, 1H), 6.72 (d, J = 8.9 Hz, 1H), 2.32 (td, J = 12.7, 4.5 Hz, 1H), 2.20 (td, J = 12.8, 4.2 Hz, 1H), 1.48 - 1.21 (m, 2H), 1.16 - 1.03 (m, 1H), 0.83 (m, 4H).

[0231] 13C NMR (CD3OD, 126 MHz) 8: 183.36, 157.93, 140.57 (q, J = 2.3 Hz), 137.36, 132.05, 129.46, 128.79, 125.58 (q, J = 4.5 Hz), 125.27 (q, J = 272.4 Hz), 123.35, 116.39, 113.35 (q, J = 33.2 Hz), 57.06, 37.84, 27.74, 23.83, 14.15.19F NMR (CD3OD, 471 MHz) 5: -64.44.

[0232] HRMS (ESI): m / z calc, for C19 H17 N 02 F3 [M-H]- 348.1211, found: 348.1205.

[0233] 3-hydroxy-3-pentyl-7-(trifluoromethyl)indolin-2-one (30a ORMPM 06 43). In an oven-dried flask 7-trifluoromethyl isatin (300 mg, 1.4 mmol, 1 eq.) was dissolved in tetrahydrofuran (10 mL) and cooled to -78° C. A IM solution of n-pentylmagnesium bromide in tetrahydrofuran (4.18 mL, 4.18 mmol, 3 eq.) was added drop-wise and reaction was stirred for 1 hour at -78° C. Reaction was quenched with saturated ammonium chloride, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 116 milligrams, 29% yield.

[0234] 'H NMR (CD3OD, 500 MHz) 8: 7.54 (d, J= 7.4 Hz, 1H), 7.50 (d, J= 8.0 Hz, 1H), 7.20 (t, J= 7.7 Hz, 1H), 2.06 - 1.86 (m, 2H), 1.23 (m, 4H), 1.14 (m, 1H), 1.03 - 0.93 (m, 1H), 0.85 - 0.78 (m, 3H).

[0235] 13C NMR (CD3OD, 126 MHz) 8: 182.06, 140.42 (q, J = 2.2 Hz), 135.03, 128.67, 126.75 (q, J = 4.4 Hz), 125.15 (q,J= 270.8 Hz), 123.50, 113.39 (q,J= 33.2 Hz), 76.59, 38.80, 32.87, 23.80, 23.33, 14.17.19F NMR (CD3OD, 471 MHz) 8: -62.94.

[0236] HRMS (ESI): m / z calc, for C14 H15 N 02 F3 [M-H]- 286.1055, found: 286.1064.

[0237] 3-(4-hydroxyphenyl)-3-pentyl-7-(trifluoromethyl)indolin-2-one (30 OR MPM 03 267). In an oven-dried flask, 30a (120 mg, .42 mmol, 1 eq.) was dissolved in dichloroethane (4 mL). Phenol was added (200 mg, 2.1 mmol, 5 eq.) followed by p-toluenesulfonic acid (542 mg, 3.15 mmol, 7.5 eq.). Reaction was heated at 90° C for 2-4 hours and then cooled to room temperature. Solid was fdtered off and crude mixture was purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 57 milligrams, 38% yield.

[0238] 'H NMR (CD3OD, 500 MHz) 8: 7.50 (d, J= 8.0 Hz, 1H), 7.38 (d, J= 7.4 Hz, 1H), 7.20 (t, J= 7.8 Hz, 1H), 7.10 (d, J = 8.8 Hz, 1H), 6.72 (d, J = 8.9 Hz, 1H), 2.31 (td, J = 12.7, 4.5 Hz, 1H), 2.19 (td, J = 12.7, 4.2 Hz, 1H), 1.39 - 1.18 (m, 4H), 1.16 - 1.08 (m, 1H), 0.97 - 0.72 (m, 4H).13C NMR (CD3OD, 126 MHz) δ: 183.37, 157.94, 140.58, 137.37, 132.05, 129.46, 128.80, 126.35, 125.58 (q, J = 4.2 Hz) , 125.28 (q, J = 269.7 Hz) , 123.35, 116.39, 113.36 (q, J = 33.2 Hz) , 57.11, 38.02, 32.95, 25.16, 23.35, 14.21.

[0239] 19F NMR (CD3OD, 471 MHz) δ: -62.93.

[0240] HRMS (ESI): m / z calc, for C20 H19 N 02 F3 [M-H]- 362.1368, found: 362.1360.

[0241] 3-(ethylamino)-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2-one (31 OR MPM 03 243). In a flame dried flask under argon, 14 (340 mg, .8 mmol, 1 eq.) was dissolved in tetrahydrofuran (7 mL) and cooled to 0° C. Pyridine (190 uL, 2.4 mmol, 3 eq.) was added dropwise followed by thionyl chloride (292 uL, 4 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0242] In a separate flame dried flask, 2 M ethyl amine (800 uL, 1.6 mmol, 2 eq.) and cesium carbonate (651 mg, 2 mmol, 2.5 eq.) were stirred in dichloromethane (5 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (8 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred for 48 hours. Reaction was quenched with water, extracted with di chloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in tetrahydrofuran (10 mL) under argon and 1 M tetrabutyl ammonium fluoride (3.2 mL, 3.2 mmol, 4 eq.) was added dropwise. Reaction stirred for 2 hours at room temperature and quenched with sodium bicarbonate, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 127 milligrams, 47% yield over 3 steps.

[0243] 'H NMR (CD3OD, 500 MHz) 8: 7.52 (d, J = 8.1 Hz, 1H), 7.49 (d, J = 7.5 Hz, 1H), 7.24 (d, J = 8.8 Hz, 2H), 7.20 (t, J= 7.8 Hz, 1H), 6.73 (d, J= 8.9 Hz, 2H), 2.44 (dq, J= 10.6, 7.1 Hz, 1H), 2.23 (dq, J = 10.8, 7.4 Hz, 1H), 1.08 (t, J= 7.1 Hz, 3H).

[0244] 13C NMR (CD3OD, 126 MHz) 8: 182.37, 158.62, 140.52 (q, J = 2.3 Hz), 135.03, 131.56, 130.19, 128.53, 126.54 (q, J = 4.5 Hz), 125.18 (q, J = 270.9 Hz), 123.55, 116.36, 113.65 (q, J = 33.2 Hz), 69.85, 39.56, 15.22.

[0245] 19F NMR (CD3OD, 471 MHz) 8: -62.90.

[0246] HRMS (ESI): m / z calc. for C17 H14 N2 02 F3 [M-H]- 335.1007, found: 335.1010.

[0247] 3-(4-hydroxyphenyl)-3-(propylamino)-7-(trifluoromethyl)indolin-2-one (32 OR

[0248] MPM 03 245). In a flame dried flask under argon, 14 (340 mg, .8 mmol, 1 eq.) was dissolved in tetrahydrofuran (7 mL) and cooled to 0° C. Pyridine (190 uL, 2.4 mmol, 3 eq.) was added dropwise followed by thionyl chloride (292 uL, 4 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0249] In a separate flame dried flask, propylamine (131 uL, 1.6 mmol, 2 eq.) and cesium carbonate (651 mg, 2 mmol, 2.5 eq.) were stirred in dichloromethane (5 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (5 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred for 48 hours. Reaction was quenched with water, extracted with di chloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in tetrahydrofuran (10 mL) under argon and 1 M tetrabutyl ammonium fluoride (3.2 mL, 3.2 mmol, 4 eq.) was added dropwise. Reaction stirred for 2 hours at room temperature and quenched with sodium bicarbonate, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 145 milligrams, 52% yield over 3 steps.

[0250] 'H NMR (CD3OD, 500 MHz) 8: 7.52 (d, J = 8.0 Hz, 1H), 7.48 (d, J = 7.5 Hz, 1H), 7.25 (d, J = 8.8 Hz, 2H), 7.20 (t, J= 7.7 Hz, 1H), 6.73 (d, J= 8.9 Hz, 1H), 2.36 (ddd, J= 10.7, 7.9, 6.4 Hz, 1H), 2.22 - 2.05 (m, 1H), 1.58 - 1.43 (m, 2H), 0.87 (t, J= 7.4 Hz, 3H).

[0251] 13C NMR (CD3OD, 126 MHz) 8: 182.54, 158.62, 140.47 (q, J = 2.2 Hz), 135.17, 131.74, 130.17, 128.50, 126.50 (q, J = 4.6 Hz), 125.18 (q, J = 270.9 Hz), 123.55, 116.36, 113.64 (q, J = 33.2 Hz), 69.81, 47.16, 24.22, 12.04.

[0252] 19F NMR (CD3OD, 471 MHz) 8: -62.87.

[0253] HRMS (ESI): m / z calc, for Cl 8 Hl 6 N2 02 F3 [M-H]- 349.1164, found: 349.1159.

[0254] 3-( 4-hydroxyphenyl)-3-(pentylamino)-7-( trifluoromethyl)indolin-2-one (33 OR

[0255] MPM 03 247). In a flame dried flask under argon, 14 (340 mg, .8 mmol, 1 eq.) was dissolved in tetrahydrofuran (7 mL) and cooled to 0° C. Pyridine (190 uL, 2.4 mmol, 3 eq.) was added dropwise followed by thionyl chloride (292 uL, 4 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0256] In a separate flame dried flask, amylamine (190 uL, 1.6 mmol, 2 eq.) and cesium carbonate (651 mg, 2 mmol, 2.5 eq.) were stirred in dichloromethane (5 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (5 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred for 48 hours. Reaction was quenched with water, extracted with di chloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in tetrahydrofuran (10 mL) under argon and 1 M tetrabutyl ammonium fluoride (3.2 mL, 3.2 mmol, 4 eq.) was added dropwise. Reaction stirred for 2 hours at room temperature and quenched with sodium bicarbonate, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 147 milligrams, 49% yield over 3 steps.

[0257] 'H NMR (CD3OD, 500 MHz) 8: 7.52 (d, J = 8.1 Hz, 1H), 7.48 (d, J = 7.5 Hz, 1H), 7.25 (d, J = 8.9 Hz, 1H), 7.20 (t, J= 7.8 Hz, 1H), 6.73 (d, J= 8.9 Hz, 1H), 2.39 (ddd, J= 10.7, 8.0, 6.4 Hz, 1H), 2.23 - 2.11 (m, 1H), 1.62 - 1.42 (m, 2H), 1.35 - 1.19 (m, 4H), 0.87 (m, 3H).

[0258] 13C NMR (CD3OD, 126 MHz) 8: 182.54, 158.63, 140.48 (q, J = 2.2 Hz), 135.15, 131.73, 130.16, 128.50, 126.49 (q, J = 4.1 Hz), 125.18 (q, J = 271.0 Hz), 123.54, 116.36, 113.65 (q, J = 33.2 Hz), 69.84, 45.18, 30.73, 30.49, 23.52, 14.29.

[0259] 19F NMR (CD3OD, 471 MHz) 8: -62.88.

[0260] HRMS (ESI): m / z calc, for C20 H20 N2 02 F3 [M-H]- 377.1477, found: 377.1471.

[0261] 3-( 4-hydroxyphenyl)-3-( ( 4, 4, 4-trifluorobutyl)amino)-7-( trifluoromethyl)indolin-2-one (34 OR MPM 04 19). In a flame dried flask under argon, 14 (200 mg, .47 mmol, 1 eq.) was dissolved in tetrahydrofuran (5 mL) and cooled to 0° C. Pyridine (114 uL, 1.42 mmol, 3 eq.) was added dropwise followed by thionyl chloride (172 uL, 2.35 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0262] In a separate flame dried flask, 4,4,4-Trifluoro-l-butanamine (119 mg, .94 mmol, 2 eq.) and cesium carbonate (383 mg, 1.18 mmol, 2.5 eq.) were stirred in di chloromethane (5 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in di chloromethane (5 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred overnight. Reaction was quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in methanol (5 mL) and 3 M potassium hydrogen difluoride in water (391 uL, 1.18 mmol, 2.5 eq.) was added dropwise. Reaction stirred for 2 hours at room temperature and quenched with sodium bicarbonate, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 85 milligrams, 43% yield over 3 steps.

[0263] 'H NMR (CD3OD, 500 MHz) 8: 7.51 (d, J = 8.0 Hz, 1H), 7.47 (d, J = 7.5 Hz, 1H), 7.27 (d, J = 8.8 Hz, 1H), 7.19 (t, J= 7.8 Hz, 1H), 6.74 (d, J= 8.8 Hz, 1H), 2.49 (m, 1H), 2.21 (m, 3H), 1.69 (m 2H).

[0264] 13C NMR (CD3OD, 126 MHz) 8: 182.42, 158.62, 140.38 (d,J = 2.2 Hz), 135.30, 131.84, 130.05, 128.9 (q, J= 273.4 Hz) , 128.52, 126.51 (q,J= 4.5 Hz), 125.17 (q, J = 271.0 Hz), 123.60, 113.66 (q, J= 33.2 Hz), 69.81, 43.82, 32.27 (q, J= 28.5 Hz), 23.74 (q, J= 3.0 Hz).

[0265] 19F NMR (CD3OD, 471 MHz) 8: -62.87, -67.94 (t, J= 11.3 Hz).

[0266] HRMS (ESI): m / z calc, for C19 H15 N2 02 F6 [M-H]- 417.1038 found: 417.1030.

[0267] 3-(azetidin-l-yl)-3-( 4-hydroxyphenyl)-7-( trifluoromethyl)indolin-2-one ( 35 OR

[0268] MPM 05 139). In a flame dried flask under argon, 14 (300 mg, .7 mmol, 1 eq.) was dissolved in tetrahydrofuran (5 mL) and cooled to 0° C. Pyridine (182 uL, 2.1 mmol, 3 eq.) was added dropwise followed by thionyl chloride (255 uL, 3.5 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0269] In a separate flame dried flask, azetidine hydrochloride (131 mg, 1.4 mmol, 2 eq.) and cesium carbonate (571mg, 1.75 mmol, 2.5 eq.) were stirred in dichloromethane (5 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (5 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred overnight. Reaction was quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in tetrahydrofuran (10 mL) under argon and 1 M tetrabutylammonium fluoride (2.1 mL, 2.1 mmol, 3 eq.) was added dropwise. Reaction stirred for 2 hours at room temperature and quenched with sodium bicarbonate, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 62 milligrams, 25% yield over 3 steps.

[0270] 'H NMR (CD3OD, 500 MHz) 8: 7.46 (m, 2H), 7.34 - 7.25 (m, 2H), 7.14 (t, J = 7.8 Hz, 1H), 6.84 - 6.68 (m, 2H), 3.47 (q, J= 7.1 Hz, 2H), 3.21 (q, J= 7.1 Hz, 2H), 2.33 - 1.85 (m, 2H).

[0271] 13C NMR (CD3OD, 126 MHz) 8: 180.40, 158.58, 140.17 (q, J = 2.3 Hz), 133.97, 130.36, 129.42, 129.40, 126.44 (q, J = 4.5 Hz), 125.15 (q, J = 270.9 Hz), 123.37, 116.41, 113.40 (q, J = 33.1 Hz), 71.93, 49.78, 17.43.

[0272] 19F NMR (CD3OD, 471 MHz) 8: -62.87.

[0273] HRMS (ESI): m / z calc, for C18 H14 N2 02 F3 [M-H]- 347.1007 found: 347.1002. General Method A (Compounds 36-44). An oven-dried 25 rnL round bottom flask was charged with 2.3 equivalents of (4-bromophenoxy)(tert-butyl) dimethylsilane, dissolved in 5 mL of THF, and cooled to -78 °C while stirring under argon gas. After cooling, 2.1 equivalents of nButyllithium solution (1.5 M) were added dropwise over 5 minutes. The reaction continued to stir for 1 hour at -78 °C. Slowly, over 20 minutes, 1 equivalent of the desired starting material (125 to 300 mg) dissolved in tetrahydrofuran (4 mL) was added to reaction and stirred for 2-3 hours at -78 °C (monitored via TLC) and then removed from the acetone-dry ice bath and allowed to warm to room temperature. The reaction was quenched with water (5 mL) and poured into a separatory funnel and extracted with ethyl acetate (3x, 20-30 mL) and a 1:1 mixture of water and brine (20-30 mL). The layers were washed with brine, then dried over Na2SO4, filtered, and concentrated in vacuo. The resultant oil was purified via automated flash chromatography using ethyl acetate / hexanes as the mobile phase to afford the corresponding tertiary alcohol 36a-44a.

[0274] Following purification, an oven-dried 15 mL round bottom flask was charged with the tertiary alcohol (1 eq.), tetrahydrofuran (3 mL) and pyridine (3 eq.) was cooled to 0 °C, stirring, under argon gas. After cooling, thionyl chloride (SOCb, 5 equivalents) and stirred until the starting material was consumed by TLC. Water (4 mL) was added to the reaction vessel. The resultant solution was then extracted with dichloromethane (3x), and the combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude oil was then dissolved in dichloromethane (2 mL). 3,3,4,4-tetrafluoropyrrolidine hydrochloride (2 eq.), di chloromethane (3mL) and cesium carbonate (3 eq.) was prepared in parallel, stirring at 0 °C. After 10 minutes, the dichlormethane-dissolved crude oil was added to the mixture containing the hydrochloride salt and stirred overnight and poured into water (10 mL). Organics were extracted with ethyl acetate (3x). The combined organic layers were washed with 1 : 1 waterbrine, dried over sodium sulfate, filtered, and concentrated in vacuo.

[0275] The resulting crude oil was then dissolved in THF (4 mL) and cooled to 0 °C, stirring, under argon gas. Tetrabutyl ammonium fluoride in tetrahydrofuran (1 M, 4 equivalents) was added and the reaction was stirred for two hours. The reaction mixture was poured into saturated sodium bicarbonate (10 mL) and extracted with ethyl acetate (3x), washed with 1: 1 brine: saturated sodium bicarbonate, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified via flash chromatography using ethyl acetate / hexanes as the mobile phase to afford the final compound 36-44. 6, 7-difluoro-3-(4-hydroxyphenyl)-3-(3,3,4,4-tetrafluoropyrrolidin-l-yl)indolin-2-one (36 or HWC 02 30). Isolated yield: 23 milligrams, 8% yield over 4 steps.

[0276] 'H NMR (CD30D, 600 MHz) 8: 7.34 (d, J= 8.7 Hz, 2H), 7.14 (ddd, J= 8.4, 4.3, 1.3 Hz, 1H), 6.93 (ddd, J= 11.1, 8.4, 7.0 Hz, 1H), 6.80 (d, J= 8.8 Hz, 2H), 3.28 (q, J= 12.7 Hz, 2H), 3.10 (q, J= 12.5 Hz, 2H).

[0277] 13C NMR (CD3OD, 151 MHz) 8: 178.35, 159.30, 152.50 (dd, J= 246.5, 9.9 Hz), 137.62 (dd, J = 247.9, 17.0 Hz), 132.41 - 131.26 (m), 129.43, 128.50, 127.80, 122.64 (dd, J= 8.1, 3.9 Hz), 119.9 (tt, 7= 261.6, 21.6 Hz), 116.85, 111.57 (d, 7= 19.0 Hz), 71.36, 54.30.

[0278] 19F NMR (CD3OD, 565 MHz) 8: -119.87 (t, 7 = 13.0 Hz), -138.84 (ddd, 7= 19.9, 11.0, 4.6 Hz), - 158.93 (dd, 7 = 19.8, 7.2 Hz).

[0279] HRMS (ESI): m / z calc, for Cl 8 Hl 1 N2 02 F6 [M-H]- 401.0725 found: 401.0730.

[0280] 7-fluoro-3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-I-yl)indolin-2-one (37 OR HWC 01 145). Isolated yield: 117 milligrams, 25% yield over 4 steps.

[0281] 'H NMR (CD3OD, 500 MHz) 8: 7.35 (d, 7= 8.9 Hz, 1H), 7.17 (dd, 7= 7.1, 1.4 Hz, 1H), 7.14 - 7.03 (m, 2H), 6.80 (d, 7 = 8.8 Hz, 2H), 3.26 (q, 7= 12.8 Hz, qH), 3.11 (q, 7= 12.5 Hz, 2H).

[0282] 13C NMR (CD3OD, 126 MHz) 8: 178.25, 159.18, 148.80 (d, 7= 244.6 Hz), 133.14 (d, 7= 2.8 Hz), 129.72 (d, 7 = 12.7 Hz), 128.59, 124.75 (d, 7= 5.8 Hz), 122.35 (d, 7= 3.4 Hz), 120.04 (tt, 7= 259.7, 23.7 Hz), 117.46 (d, 7 = 17.3 Hz), 116.74, 71.59, 54.29 (t, 7= 27.9 Hz).

[0283] 19F NMR (CD3OD, 471 MHz) 8: -119.85 (t, 7 = 13.0 Hz), -134.78 (dd, 7= 9.6, 5.2 Hz).

[0284] HRMS (ESI): m / z calc, for C18 H12 N2 02 F5 [M-H]- 383.0819 found: 383.081.

[0285] 5, 6, 7-trifluoro-3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-l-yl)indolin-2-one ( 38

[0286] ORMPM 05 275). Isolated yield: 70 milligrams, 7% yield over 4 steps.

[0287] 'H NMR (CD3OD, 500 MHz) 8: 7.41 - 7.30 (m, 2H), 7.21 (m, 1H), 6.85 - 6.77 (m, 2H), 3.45 (q, 7 = 12.3 Hz, 2H), 3.10 (q, 7 = 12.5 Hz, 2H).

[0288] 13C NMR (CD3OD, 126 MHz) 8: 177.94, 159.39, 148.64 (dd, 7= 244.5, 10.9 Hz), 145.33 - 140.36 (m), 138.18 (dd, J= 249.0, 13.6 Hz), 129.35, 128.03, 127.26 (d, J= 9.9 Hz), 126.22, 116.92, 110.99 (dd, J= 20.8, 3.7 Hz), 71.41, 54.17 (t, J = 26.9 Hz).

[0289] 19F NMR (CD3OD, 471 MHz) δ: -119.95 (t, J= 13.1 Hz), -144.74 (dd, J= 19.0, 8.9 Hz), -155.12 (d, J= 18.9 Hz), -161.60 (d, J= 6.7 Hz).

[0290] HRMS (ESI): m / z calc, for C18 H14 N2 02 F3 [M-H]- 347.1007 found: 347.1002.

[0291] 5, 7-dichloro-3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-I-yl)indolin-2-one ( 39 OR HWC 02 34). Isolated yield: 67 milligrams, 13% yield over 4 steps.

[0292] 'H NMR (CD3OD, 500 MHz) 8: 7.46 (m, 2H), 7.34 - 7.25 (m, 2H), 7.14 (t, J = 7.8 Hz, 1H), 6.84 - 6.68 (m, 2H), 3.47 (q, J= 7.1 Hz, 2H), 3.21 (q, J= 7.1 Hz, 2H).

[0293] 13C NMR (CD3OD, 126 MHz) 8: 178.01, 159.43, 139.11, 133.63, 130.26, 129.65, 129.32, 128.08,

[0294] 125.14, 119.88 (tt, J= 260.4, 24.9 Hz), 117.47, 116.95, 72.18, 54.26(t, J= 28.6 Hz),

[0295] 19F NMR (CD3OD, 471 MHz) 8: -119.94 (t, J = 13.0 Hz).

[0296] HRMS (ESI): m / z calc, for C18 Hl l N2 02 C12 F4 [M-H]- 433.0134 found: 433.0132.

[0297] 7-fluoro-3-( 4-hydroxyphenyl)-5-nitro-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-I-yl)indolin-2-one ( 40 OR HWC 02 13). Isolated yield: 12 milligrams, 2% yield over 4 steps.

[0298] 'H NMR (CD3OD, 500 MHz) 8: 8.10 (dd, J= 9.9, 2.1 Hz, 1H), 8.06 (d, J = 2.1 Hz, 1H), 7.39 (d, J = 8.9 Hz, 2H), 6.84 (d, J= 8.8 Hz, 2H), 3.41 (q, J= 12.6 Hz, 2H), 3.14 (q, J= 12.5 Hz, 2H).

[0299] 13C NMR (CD3OD, 126 MHz) 8: 178.42, 159.64, 147.44 (d, J = 248.7 Hz), 144.97 (d, J = 6.3 Hz), 136.41 (d, J= 13.3 Hz), 133.70 (d, J = 3.8 Hz), 129.35, 127.63, 119.76 (tt, J= 261.8, 23.9 Hz), 117.88, 117.11 , 114.47 (d, J = 22.6 Hz), 71.49, 54.22 (t, J = 28.1 Hz).

[0300] 19F NMR (CD3OD, 471 MHz) 8: -120.05 (td, J= 13.2, 3.2 Hz), -131.90 (d, J= 10.2 Hz).

[0301] HRMS (ESI): m / z calc, for C18 Hl l N3 04 F5 [M-H]- 428.0670 found: 428.0672.

[0302] 5, 7-dibromo-3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-I-yl)indolin-2-one (41 OR MPM 05 235). Isolated yield: 140 milligrams, 37% yield over 4 steps.

[0303] 'H NMR (CD3OD, 500 MHz) 8: 7.61 (d, J = 1.8 Hz, 1H), 7.45 (d, J = 1.8 Hz, 1H), 7.34 (d, J = 8.8 Hz, 2H), 6.82 (d, J = 8.8 Hz, 2H), 3.35 (q, J = 12.3 Hz, 2H), 3.18 - 3.03 (q, J = 12.5 Hz, 2H).

[0304] 13C NMR (CD3OD, 126 MHz) 8: 177.83, 159.42, 141.25, 135.68, 133.93, 129.30, 128.31, 128.14, 119.87 (tt, J = 260.2, 23.6 Hz), 116.97, 116.54, 105.11, 72.38, 54.26 (t, J= 27.9 Hz).

[0305] 19F NMR (CD3OD, 471 MHz) 8: -119.93 (t, J= 13.1 Hz).

[0306] HRMS (ESI): m / z calc, for C18 Hl l N2 02 Br2 F4 [M-H]- 520.9123 found: 520.9121.

[0307] 3-( 4-hydroxyphenyl)-2-oxo-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-l-yl)indoline-5-carbonitrile ( 42 OR HWC 02 48).

[0308] 'H NMR (CD3OD, 500 MHz) 8: 7.72 (d, J= 1.7 Hz, 1H), 7.66 (dd, J= 8.2, 1.7 Hz, 1H), 7.49 - 7.32 (m, 2H), 7.07 (d, J = 8.1 Hz, 1H), 6.82 (q, J = 8.9 Hz, 2H), 3.36 (q, J = 12.7 Hz, 2H), 3.11 (q, J = 12.5 Hz, 2H).

[0309] 13C NMR (CD3OD, 126 MHz) 8: 178.51, 159.42, 146.71, 135.78, 131.83, 130.00, 129.37, 128.07, 119.89 (tt, J= 259.9, 24.8 Hz), 119.79, 116.95, 112.48, 107.08, 71.06, 54.24 (t, J = 28.0 Hz).

[0310] 19F NMR (CD3OD, 471 MHz) 8: -119.98 (t, J= 13.1 Hz).

[0311] HRMS (ESI): m / z calc, for C19 H12 N3 02 F4 [M-H]- 390.0866 found: 390.0866.

[0312] 5-bromo-3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-l-yl)-7-( trifluoro- methyl)indolin-2-one (43 OR HWC 02 17). Isolated yield: 43 milligrams, 10% yield over 4 steps. 'H NMR (CD3OD, 500 MHz) δ: 7.73 (d, J = 2.0 Hz, 1H), 7.69 (d, J = 2.0 Hz, 1H), 7.37 - 7.28 (m, 2H), 6.83 (d, J= 8.8 Hz, 1H), 3.38 (q, J= 12.7 Hz, 2H), 3.11 (q, J= 12.5 Hz, 2H).

[0313] 13C NMR (CD3OD, 126 MHz) δ: 178.27, 159.51, 138.96, 135.03, 132.94, 129.87 (q, J = 4.8 Hz), 127.93, 124.04 (q, J = 271.7 Hz), 119.85 (tt, J= 264.4, 23.0 Hz ), 117.05, 116.01, 115.49 (q, J= 33.9 Hz), 70.28, 54.22 (t, J = 28.1 Hz).

[0314] 19F NMR (CD3OD, 471 MHz) δ: -63.32, -119.98 (t, J= 13.2 Hz).

[0315] HRMS (ESI): m / z calc, for C19 Hl l N2 02 F7 Br [M-H]- 510.9892 found: 510.9901.

[0316] 3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-l-yl)-l, 3-dihydro-2H-pyrrolo[2, 3- b]pyridin-2-one (44 OR JC 01 12). Isolated yield: 53 milligrams, 18% yield over 4 steps.

[0317] 1H NMR (CD3OD, 500 MHz) 8: 8.35 - 8.04 (m, 1H), 7.73 (dd, J= 7.5, 1.7 Hz, 1H), 7.36 (d, J= 8.8 Hz, 2H), 7.06 (dd, J = 7.5, 5.2 Hz, 1H), 6.81 (d, J= 8.8 Hz, 2H), 3.31 (q, J = 12.5 Hz, 2H), 3.11 (q, J = 12.5 Hz, 2H).

[0318] 13C NMR (CD3OD, 126 MHz) 8: 178.20, 159.33, 157.02, 149.18, 134.87, 129.39, 128.03, 125.11, 119.91, 119.88 (tt, J= 262.9, 23.5 Hz), 116.88, 71.38, 54.39 (t, J = 27.9 Hz).

[0319] 19F NMR (CD3OD, 471 MHz) 8: -120.61 (t, J= 13.1 Hz).

[0320] HRMS (ESI): m / z calc, for C17 H14 N3 02 F4 [M-H]- 368.1022 found: 368.1020.

[0321] 7-bromo-3-(4-((tert-butyldimethylsilyl)oxy)phenyl)-3-hydroxyindolin-2-one (45a OR MPM 05 209). In an oven-dried flask (4-bromophenoxy)(tert-butyl) dimethylsilane (5 mL, 20 mmol, 2.3 eq.) was dissolved in 20 mL of tetrahydrofuran, and cooled to -78 °C while stirring under argon gas. After cooling, 1.6 M wButyllithium in hexanes (11.7 mL, 18.7 mmol, 2.1 eq.) was added dropwise. The reaction was stirred at -78 °C for 30 minutes followed by addition of 7-bromo isatin (2 g, 8.9 mmol, 1 eq.) dissolved in tetrahydrofuran (5 mL). The reaction continued to stir at -78 °C for 1 hr, and then allowed to warm to room temperature. Reaction was quenched with water, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 2.05 grams, 54% yield.

[0322] 'H NMR (CD3OD, 500 MHz) 8: 7.44 (dd, J= 8.2, 1.1 Hz, 1H), 7.30 - 7.23 (m, 2H), 7.17 (dd, J= 7.4, 1.2 Hz, 1H), 7.04 - 6.95 (m, 1H), 6.87 - 6.76 (m, 2H), 0.98 (s, 9H), 0.18 (s, 6H).

[0323] 13C NMR (CD3OD, 126 MHz) 8: 181.11, 156.99, 142.55, 136.56, 134.34, 133.48, 128.12, 125.32, 125.06, 120.96, 104.04, 79.72, 26.11, 19.04, -4.37.

[0324] HRMS (ESI): m / z calc, for C20 H23 N 03 Si Br [M-H]- 432.0631 found: 432.0635.

[0325] 7-bromo-3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-I-yl)indolin-2-one ( 45 OR MPM 05 239). In an oven dried flask under argon, 45a (1.2 g, 2.8 mmol, 1 eq.) was dissolved in tetrahydrofuran (20 mL) and cooled to 0° C. Pyridine (712 uL, 8.3 mmol, 3 eq.) was added dropwise followed by thionyl chloride (1 mL, 13.8 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0326] In a separate flame dried flask, 3,3,4,4-tetrafluoropyrrolidine hydrochloride (700 mg, 5.5 mmol, 2 eq.) and cesium carbonate (2.25 g, 6.9 mmol, 2.5 eq.) were stirred in di chloromethane (20 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (5 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred for 48 hours. Reaction was quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with NaiSCL, filtered and concentrated. Crude mixture was dissolved in tetrahydrofuran (30 mL) under argon and 1 M tetrabutylammonium fluoride (8.2 mL, 8.2 mmol, e eq.) was added dropwise. Reaction stirred for 1 hours at room temperature and quenched with ammonium chloride, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSCL, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 660 milligrams, 54% yield.

[0327] 'H NMR (CD3OD, 500 MHz) 8: 7.43 (d, J = 8.1 Hz, 1H), 7.35 (d, J = 8.7 Hz, 2H), 7.31 (d, J = 7.5 Hz, 1H), 6.99 (t, J= 7.8 Hz, 1H), 6.80 (d, J= 8.6 Hz, 2H), 3.28 (q, J= 12.7 Hz, 2H), 3.11 (q, J= 12.5 Hz, 2H).

[0328] 13C NMR (CD3OD, 126 MHz) 8: 178.23, 159.18, 141.85, 133.56, 132.11, 129.36, 128.60, 125.43, 125.28, 123.21 - 117.33 (tt, J =261.3, 25.0 Hz), 116.78, 104.53, 72.24, 54.29 (t, J = 27.8 Hz).

[0329] 19F NMR (CD3OD, 471 MHz) δ: -119.82 (t, J = 12.9 Hz).

[0330] HRMS (ESI): m / z calc, for C18 H12 N2 02 F4 Br [M-H]- 443.0018 found: 443.0015.

[0331] (R)-7-bromo-3-(4-hydroxyphenyl)-3-(3, 3, 4, 4-tetrafluoropyrrolidin-I-yl)indolin-2-one ((R)-45 orMPM 05 263A). Enantiomers of 45 were separated on CHIRALPAK AD-H column. 25 milligrams of 45 was dissolved in 1 mL 50% isopropanol / hexanes, fdtered, and separated on ACCQ Prep HP125 HPLC using 7% isopropanol / hexanes as the mobile phase.

[0332] 'H NMR (CD3OD, 500 MHz) 8: 7.48 - 7.45 (m, 1H), 7.38 (d, J= 8.8 Hz, 2H), 7.34 (dd, J = 7.5, 1.0 Hz, 1H), 7.03 (t, J= 7.8 Hz, 1H), 6.83 (d, J= 8.7 Hz, 2H), 3.31 (q, J= 12.8 Hz, 2H), 3.14 (q, J= 12.5 Hz, 2H).

[0333] 13C NMR (CD3OD, 126 MHz) 8: 178.23, 159.20, 141.87, 133.57, 132.14, 129.37, 128.62, 125.45,

[0334] 125.28, 119.93 (tt, 7 = 260.8, 23.5 Hz), 116.78, 104.53, 72.24, 54.31 (t, J= 27.8 Hz).

[0335] 19F NMR (CD3OD, 471 MHz) 8: -119.85 (t, J= 12.9 Hz).

[0336] HRMS (ESI): m / z calc, for C18 H12 N2 02 Br F4 [M-H]- 443.0018 found: 443.0018.

[0337] (S)-7-bromo-3-(4-hydroxyphenyl)-3-(3,3,4,4-tetrafluoropyrrolidin-l-yl)indolin-2-one ((S)-45 orMPM 05 263B).

[0338] 'H NMR (CD3OD, 500 MHz) 8: 7.43 (dd, J= 8.2, 1.1 Hz, 1H), 7.35 (d, J= 8.8 Hz, 2H), 7.31 (dd, J = 7.5, 1.0 Hz, 1H), 6.99 (t, J= 7.9 Hz, 1H), 6.80 (d, J= 8.8 Hz, 2H), 3.30 - 3.25 (q, J= 12.4 Hz, 2H), 3.11 (q, J= 12.5 Hz, 2H).

[0339] 13C NMR (CD3OD, 126 MHz) 8: 178.23, 159.19, 141.87, 133.57, 132.14, 129.37, 128.62, 125.45,

[0340] 125.28, 119.93 (tt, 7= 260.3, 23.4 Hz), 116.78, 104.53, 72.24, 54.30 (t, J= 27.9 Hz).

[0341] 19F NMR (CD3OD, 471 MHz) 8: -119.84 (t, 7= 13.7 Hz).

[0342] HRMS (ESI): m / z calc, for C18 H12 N2 02 F4 Br [M-H]- 443.0018 found: 443.0014. 3-(4-hydroxyphenyl)-7-(propylamino)-3-(3, 3, 4, 4-tetrafluoropyrrolidin-l-yl)indolin-2-one (46 ORMPM 05 245). A flame dried flask was charged with 45 (50 mg, .11 mmol, 1 eq.), BrettPhos Pd G3 (10 mg, 0.01 mmol, 0.1 eq.), and BrettPhos (6 mg, 0.01 mmol, 0.1 eq.) and placed under vacuum before refilling with argon (3x). Propylamine (11 uL, .14 mmol, 1.2 eq.) was added followed by 1 M LiHMDS in tetrahydrofuran (330 uL, .33 mmol, 3 eq.). Reaction was heated at 80° C for 16 hours. Reaction was extracted with water / ethyl acetate, organic layers combined, dried with MgSCb, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 8 milligrams, 17% yield.

[0343] 'H NMR (CD3OD, 500 MHz) 8: 7.34 (d, J= 8.8 Hz, 2H), 6.96 (t, J= 7.9 Hz, 1H), 6.77 (d, J= 8.7 Hz, 2H), 6.67 - 6.60 (m, 2H), 3.25 (q, J = 11.6 Hz, 1H), 3.18 - 3.01 (m, 4H), 1.69 (h, J = 7.3 Hz, 2H), 1.04 (t, J = 7.4 Hz, 3H).

[0344] 13C NMR (CD3OD, 126 MHz) 8: 178.66, 158.87, 135.09, 129.71, 129.19, 128.02, 125.09, 120.06 (tt, J =258.8, 23.9 Hz), 116.51, 114.58, 112.66, 71.90, 54.34 (t, J= 27.7 Hz), 46.70, 23.50, 11.98.19F NMR (CD3OD, 471 MHz) -119.74 (m).

[0345] HRMS (ESI): m / z calc, for C21 H20 N3 02 F4 [M-H]- 422.1492 found: 422.1491.

[0346] 7-(butylamino)-3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-l-yl)indolin-2-one (47 OR MPM 05 249). A flame dried flask was charged with 45 (90 mg, .2 mmol, 1 eq.), BrettPhos Pd G3 (36 mg, 0.04 mmol, 0.2 eq.), and BrettPhos (11 mg, 0.02 mmol, 0.1 eq.) and placed under vacuum before refilling with argon (3x). Butylamine (24 uL, .24 mmol, 1.2 eq.) was added followed by 1 M LiHMDS in tetrahydrofuran (600 uL, .6 mmol, 3 eq.). Reaction was heated at 60° C for 2 hours. Reaction was extracted with water / ethyl acetate, organic layers combined, dried with MgSCL, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 22 milligrams, 25% yield.

[0347] 'H NMR (CD3OD, 500 MHz) 8: 7.34 (d, J= 8.8 Hz, 2H), 6.96 (t, J= 7.0 Hz, 1H), 6.77 (2, J= 8.8 Hz, 1H), 6.63 (m, 2H), 3.25 (q, J= 12.0 Hz, 2H), 3.15 (t, J= 7.1 Hz, 2H), 3.13 - 3.05 (m, 2H), 1.66 (tt, J= 8.6, 6.8 Hz, 2H), 1.55 - 1.41 (m, 2H), 0.99 (t, J= 7.3 Hz, 3H).

[0348] 13C NMR (CD3OD, 126 MHz) 8: 178.66, 158.87, 135.12, 129.70, 129.19, 128.01, 125.10, 120.06 (t, J= 260.7, 23.4 Hz), 116.51, 114.58, 112.64, 71.90, 54.34 (t, J= 27.8 Hz), 44.55, 32.53, 21.36, 14.26.19F NMR (CD3OD, 471 MHz) 8: -119.74 (m). HRMS (ESI): m / z calc, for C22 H22 N3 02 F4 [M-H]- 436.1648 found: 436.1646.

[0349] 7-(hexylamino)-3-( 4-hydroxyphenyl)-3-( 3, 3, 4, 4-tetrafluoropyrrolidin-I-yl)indolin-2-one ( 48 ORMPM 05 253). A flame dried flask was charged with 45 (130 mg, .29 mmol, 1 eq.), BrettPhos Pd G3 (79 mg, 0.09 mmol, 0.3 eq.), and BrettPhos (16 mg, 0.03 mmol, 0.1 eq.) and placed under vacuum before refilling with argon (3x). Hexylamine (50 uL, .35 mmol, 1.2 eq.) was added followed by 1 M LiHMDS in tetrahydrofuran (870 uL, .87 mmol, 3 eq.). Reaction was heated at 60° C for 48 hours. Reaction was extracted with water / ethyl acetate, organic layers combined, dried with MgSCb, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Separation by reverse phase HPLC was required: 30% acetonitrile to 100% acetonitrile over 25 minutes. Isolated yield: 5 milligrams, 4% yield.

[0350] 'H NMR (CD3OD, 500 MHz) 8: 7.34 (d, J = 8.8 Hz, 2H), 6.97 (t, J = 7.8 Hz, 1H), 6.77 (d, J = 8.8 Hz, 2H), 6.69 - 6.61 (m, 2H), 3.25 (q, J = 12.9 Hz, 2H), 3.15 (t, J = 7.1 Hz, 2H), 3.08 (q, J = 12.1 Hz, 2H), 1.76 - 1.60 (m, 2H), 1.52 - 1.42 (m, 2H), 1.36 (td, J= 4.0, 2.2 Hz, 4H), 1.08 - 0.83 (m, 3H).

[0351] 13C NMR (CD3OD, 126 MHz) 8:13C NMR (126 MHz, MeOD) 8 178.66, 158.87, 135.11, 129.71, 129.19, 128.04, 125.10, 120.06 (tt, J = 260.8, 22.9 Hz), (, 116.51, 114.60, 112.71, 71.91, 54.34 (t, J = 27.7 Hz), 44.89, 32.88, 30.35, 27.97, 23.70, 14.39.

[0352] 19F NMR (CD3OD, 471 MHz) 8: -119.75 (m).

[0353] HRMS (ESI): m / z calc, for C24 H26 N3 02 F4 [M-H]- 464.1961 found: 464.1954.

[0354] 4-(2-oxo-3-(3, 3, 4, 4-tetrafluoropyrrolidin-I-yl)-7-(trifluoromethyl)indolin-3-yl)phenyl acetate (49 ORMPM 06 37). In an oven dried flask, 26 (265 mg, .46 mmol, 1 eq.) and pyridine (149 uL, 1.84 mmol, 4 eq.) were dissolved in di chloromethane (5 mL) and cooled to 0° C. Acetic anhydride (130 uL, 1.38 mmol, 3 eq.) was added and the reaction was allowed to warm to room temperature and stirred for 3 hours. Reaction was quenched with ammonium chloride, extracted with di chloromethane (3x), organic layers were combined and dried with MgSO4 filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 180 milligrams, 62% yield.

[0355] ’H NMR (500 MHz, CDCl3) 8: 8.29 (s, 1H), 7.59 (d, J = 9.2, 2H), 7.49 (m, 2H), 7.18 (t, J = 7.8 Hz, 1H), 7.14 (d, J= 8.6 Hz, 2H), 3.39 (q, J= 12.3 Hz, 2H), 3.13 (q, J= 12.2 Hz, 2H), 2.30 (s, 3H).

[0356] 13C NMR (126 MHz, CDCl3) 8: 176.11, 169.32, 151.25, 137.69 - 136.63 (m), 133.73, 130.57, 129.05, 128.12, 126.75 (q, J= 4.0 Hz), 123.64 (q, J = 273.5 Hz), 123.64, 122.65, 118.33 (tt, J = 258.6, 23.5 Hz), 113.22 (q, J = 33.4 Hz), 69.11, 53.44 (t, J= 28.1 Hz), 21.25.

[0357] 19F NMR (471 MHz, CDCh) 8: -60.64, -118.24 (dt, J= 50.5, 12.6 Hz).

[0358] HRMS (ESI): m / z calc, for C21 H14 N2 03 F7 [M-H]- 475.0893 found: 475.0898

[0359] (R)-4-(2-oxo-3-(3, 3, 4, 4-tetrafluoropyrrolidin-l-yl)-7-(trifluoromethyl)indolin-3-yl)phenyl acetate ((R)49 OR MPM 06 37A). Enantiomers of 49 were separated on CHIRALPAK AD-H column. 50 milligrams of 49 was dissolved in 3 mL 50% isopropanol / hexanes, filtered, and separated on ACCQ Prep HP125 HPLC using 6% isopropanol / hexanes as the mobile phase.

[0360] ’H NMR (500 MHz, CDCh) 8: 8.24 (s, 1H), 7.58 (d, J = 8.8 Hz, 2H), 7.49 (m, 2H), 7.22 - 7.16 (m, 1H), 7.14 (d, J= 8.7 Hz, 2H), 3.39 (q, J= 11.8 Hz, 2H), 3.13 (q, J= 11.4 Hz, 2H), 2.30 (s, 3H).

[0361] 13C NMR (126 MHz, CDCh) 8: 176.07, 169.32, 151.25, 137.11 (q,J=2.3 Hz), 133.73, 130.57, 129.06,

[0362] 128.11, 126.74 (q, J = 4.0 Hz), 123.65, 126.64 (q, J = 271.9 Hz), 122.66, 118.33 (t, J = 258.5, 23.5 Hz), 113.20 (q, J= 33.4 Hz), 69.11, 53.44 (t, J= 28.0 Hz), 21.26.

[0363] 19F NMR (471 MHz, CDCh) 8: -60.63, -118.23 (m).

[0364] HRMS (ESI): m / z calc, for C21 H14 N2 03 F7 [M-H]- 475.0893 found: 475.0899.

[0365] (S)-4-(2-oxo-3-(3, 3, 4, 4-tetrafluoropyrrolidin-l-yl)-7-(trifluoromethyl)indolin-3-yl)phenyl acetate ((S)49 ORMPM 06 37B).

[0366] ’H NMR (500 MHz, CDCh) 8: 8.33 (s, 1H), 7.59 (d, J = 8.7 Hz, 2H), 7.49 (m, 2H), 7.18 (t, J = 7.7 Hz, 1H), 7.14 (d, J= 8.7 Hz, 2H), 3.40 (q, J= 11.6 Hz, 1H), 3.13 (q, J= 11.6 Hz, 2H), 2.30 (s, 3H).

[0367] 13C NMR (126 MHz, CDCl3) 8: 176.16, 169.32, 151.25, 137.13 (q, J = 2.0 Hz), 133.74, 130.57, 129.05,

[0368] 128.12, 126.75 (d, J = 4.0 Hz), 123.64, 123.63(q, J = 274.3 Hz)„ 122.65, 118.33 (tt, J = 260.6, 23.5 Hz), 113.23 (q, J= 33.5 Hz), 69.12, 53.44 (t, J= 28.0 Hz), 21.25.

[0369] 19F NMR (471 MHz, CDCl3) 8: -60.65, -118.26 (m). HRMS (ESI): m / z calc, for C21 H14 N2 03 F7 [M-H]- 475.0877 found: 475.0896.

[0370] 3-(4-((tert-butyldimethylsilyl)oxy)phenyl)-7-(trifluoromethyl)indolin-2-one (50 OR MPM 05 63). Compound 14 (1 g, 2.4 mmol, 1 eq.) was dissolved in neat acetic acid (12 mL) and tin (II) chloride (1.12 g, 5.9 mmol, 2.5 eq.) was added to the flask. Reaction was stirred at 80° C for 1 hour. Mixture was cooled to room temperature and concentrated, diluted with ethyl acetate, and washed with sodium bicarbonate. Crude mixture was purified via flash chromatography using ethyl acetate / hexanes as the mobile phase.

[0371] Isolated yield: 530 milligrams, 55% yield.

[0372] ’H NMR (500 MHz, CDCl3) 8: 7.88 (s, 1H), 7.46 (d, J = 8.1 Hz, 1H), 7.30 (d, J = 7.4 Hz, 1H), 7.13 (t, J= 7.8 Hz, 1H), 7.06 (d, J= 8.5 Hz, 2H), 6.81 (d, J= 8.6 Hz, 2H), 4.58 (s, 1H), 0.97 (s, 8H), 0.19 (d, 7 = 2.0 Hz, 6H).

[0373] 13C NMR (126 MHz, CDCh) 8:13C NMR (126 MHz, CDCh) 8 177.23, 155.65, 138.84, 131.44, 129.57, 128.98, 128.00, 125.29 (q, J= 4.2 Hz), 124.05 (q, J= 271.8 Hz), 120.75, 112.19 (q, J= 33.3 Hz), 50.86, 25.78, 18.31, -4.28.

[0374] 19F NMR (471 MHz, CDCh) 8: -60.72.

[0375] HRMS (ESI): m / z calc, for C21 H25 N 02 F3 Si [M+H]+ 408.1607 found: 408.1613. tert-butyl 4-((3-(4-hydroxyphenyl)-2-oxo-7-(trifluoromethyl)indolin-3-yl)methyl)piperidine-l- carboxylate (51 ORMPM 05 69). In an oven dried flask, compound 50 (112 mgs, .25 mmol, 1 eq.) was dissolved in dimethylformamide (2 mL) and the reaction was cooled to -40° C. Potassium tert- butoxide (30 mgs, 0.27 mmol, 1.2 eq.) was added and the reaction was allowed to warm to -14° C (yellow to red color transition) over the course of 5 minutes. Reaction was cooled to -50° C and tertbutyl 4-(iodomethyl)piperidine-l -carboxylate (98 mgs, .3 mmol, 1.2 eq) was added dropwise in dimethylformamide (1 mL). Reaction was allowed to warm to room temperature over the course of 4 hours. Reaction quenched with ammonium chloride, extracted with ethyl acetate (3x), the organic layers were combined, dried with MgSO4, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. The intermediate was dissolved in methanol (3 mL) at room temperature and 3M potassium bifluoride in water (100 uL, .33 mmol, 2.5 eq.) was added dropwise and reaction monitored by TLC. Upon disappearance of starting material, methanol was removed, and reaction was purified by flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 58 milligrams, 43% yield.

[0376] ’H NMR (500 MHz, MeOD) 8: 7.53 (d, J= 8.0 Hz, 1H), 7.47 (d, J= 7.4 Hz, 1H), 7.22 (t, J= 7.7 Hz, 1H), 7.11 (d, J= 8.9 Hz, 2H), 6.71 (d, J= 8.8 Hz, 2H), 3.92 - 3.78 (m, 2H), 2.52 (s, 2H), 2.38 (dd, J = 14.0, 6.5 Hz, 1H), 2.24 (dd, J= 14.0, 5.3 Hz, 1H), 1.54 (dt, J= 13.2, 3.0 Hz, 1H), 1.41 (s, 9H), 1.31 - 1.19 (m, 2H), 1.18 - 1.07 (m, 1H), 1.06 - 0.95 (m, 1H).

[0377] 13C NMR (126 MHz, MeOD) 8: 183.35, 157.99, 156.34, 140.49, 136.86, 132.70, 130.12, 128.62, 125.82 (q, J= 4.7 Hz), 125.23 (q, J = 270.5 Hz), 123.32, 116.38, 113.56 (d, J= 33.2 Hz), 80.90, 56.24, 44.49, 34.51, 33.87, 28.65.

[0378] 19F NMR (471 MHz, MeOD) 8: -63.73.

[0379] HRMS (ESI): m / z calc, for C26 H28 N2 04 F3 [M-H]- 489.2001 found: 489.2010.

[0380] 3-(4-hydroxyphenyl)-3-(piperidin-4-ylmethyl)-7-(trifluoromethyl)indolin-2-one (52 OR MPM 05 109). Compound 51 (40 mgs, .08 mmol) was dissolved in di chloromethane (2 mL) at room temperature and trifluoroacetic acid (100 uL) was added dropwise. Reaction was monitored by TLC and upon completion, dichloromethane was removed by rotary evaporation and mixture purified via flash chromatography using 10-20% methanol / di chloromethane as the mobile phase. Isolated yield: 34 milligrams, quantitative.

[0381] ’H NMR (500 MHz, MeOD) 8: 7.55 (d, J= 8.1 Hz, 1H), 7.51 (d, J= 7.5 Hz, 1H), 7.24 (t, J= 7.8 Hz, 1H), 7.13 (d, J= 8.9 Hz, 2H), 6.72 (d, J= 8.9 Hz, 2H), 3.23 (ddt, J= 30.7, 12.9, 2.9 Hz, 2H), 2.77 (qd, J= 11.0, 4.9 Hz, 2H), 2.44 (dd, J= 14.5, 5.1 Hz, 1H), 2.29 (dd, J= 14.5, 3.9 Hz, 1H), 1.85 - 1.75 (m, 1H), 1.60 - 1.20 (m, 4H).

[0382] 13C NMR (126 MHz, MeOD) 8: 183.00, 158.12, 140.38, 136.40, 132.24, 130.11, 128.60, 126.04 (q, J = 4.4 Hz), 125.17 (q, J = 270.43 Hz), 123.50, 116.46, 113.69 (q, J = 33.2 Hz), 56.02, 44.71, 43.78, 32.20, 31.14, 30.50.

[0383] 19F NMR (471 MHz, MeOD) 5: -63.69.

[0384] HRMS (ESI): m / z calc, for C21 H20 N2 02 F3 [M-H]- 389.1477 found: 389.1473. tert-butyl 4-(3-(4-hydroxyphenyl)-2-oxo-7-(trifluoromethyl)indolin-3-yl)piperidine-l- carboxylate (53 ORMPM 05 91). In an oven dried flask, compound 50 (310 mgs, .76 mmol, 1 eq.) was dissolved in dimethylformamide (6 mL) and the reaction was cooled to -40° C. Potassium tert- butoxide (93 mgs, 0.83 mmol, 1.1 eq.) was added and the reaction was allowed to warm to -14° C (yellow to red color transition) over the course of 5 minutes. Reaction was cooled to -50° C and tertbutyl 4-iodopiperidine-l -carboxylate (472 mgs, 1.5 mmol, 2 eq) was added dropwise in dimethylformamide (1 mL). Reaction was allowed to warm to room temperature over the course several hours and left overnight. Reaction quenched with ammonium chloride, extracted with ethyl acetate (3x), the organic layers were combined, dried with MgSO4, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase to afford the TBS-deprotected compound. Isolated yield: 90 milligrams, 25% yield.

[0385] ’H NMR (500 MHz, MeOD) 8: 7.58 (d, J = 7.5 Hz, 1H), 7.54 (d, J = 8.0 Hz, 1H), 7.27 - 7.18 (m, 3H),

[0386] 6.75 (d, J= 8.9 Hz, 2H), 4.13 - 3.92 (m, 2H), 2.77 - 2.59 (m, 3H), 1.59 - 1.43 (m, 2H), 1.39 (m, 10H), 1.12 - 0.91 (m, 1H).

[0387] 13C NMR (126 MHz, MeOD) 8: 179.79, 155.93, 154.22, 138.63, 132.24, 128.67, 127.38, 127.03,

[0388] 123.75 (q, J= 4.4 Hz), 123.08 (q, J = 270.7 Hz), 120.79, 114.35, 111.28 (q, J= 33.2 Hz), 78.90, 57.97, 43.01, 26.48, 25.79, 25.65.

[0389] 19F NMR (471 MHz, MeOD) 8: -63.79.

[0390] HRMS (ESI): m / z calc, for C25 H26 N2 04 F3 [M-H]- 475.1845 found: 475.1858.

[0391] 3-(4-hydroxyphenyl)-3-(piperidin-4-yl)-7-(trifluoromethyl)indolin-2-one (54 OR MPM 05 111). Compound 53 (45 mgs, .1 mmol) was dissolved in dichloromethane (4 mL) at room temperature and trifluoroacetic acid (100 uL) was added dropwise. Reaction was monitored by TLC and upon completion, dichloromethane was removed by rotary evaporation and mixture purified via flash chromatography using 10-20% methanol / di chloromethane as the mobile phase. Isolated yield: 10 milligrams, 28% yield.

[0392] ’H NMR (500 MHz, CDCl3) 8: 7.61 (d, J= 7.5 Hz, 1H), 7.55 (d, J= 8.1 Hz, 1H), 7.25 (t, J= 7.9 Hz, 1H), 7.22 - 7.19 (m, 2H), 6.76 - 6.70 (m, 2H), 3.04 (dd, J= 29.8, 12.6 Hz, 2H), 2.70 - 2.53 (m, 3H), 1.68 - 1.39 (m, 3H), 1.25 - 0.99 (m, 1H).

[0393] 13C NMR (126 MHz, CDCl3) 8: 182.01, 158.15, 140.87, 134.36, 131.01, 129.54, 129.05, 125.85 (q, J = 4.6 Hz), 125.23 (d, J = 272.8 Hz), , 122.90, 116.50, 113.38 (d, J= 33.4 Hz), 60.35, 47.10, 46.77, 44.91, 28.02, 27.76.

[0394] 19F NMR (471 MHz, CDCh) 8: -63.78.

[0395] HRMS (ESI): m / z calc, for C20 H18 N2 02 F3 [M-H]- 375.1320 found: 375.1312.

[0396] Scheme 10. Synthesis of Compounds 55-57. Scheme 11. Synthesis of Compounds 58-61.

[0397] Scheme 12. Synthesis of Compounds 62, 63, & 65.

[0398] Scheme 13. Synthesis of Compounds 64.

[0399] Scheme 14. Synthesis of Compounds 66.

[0400] 3-hydroxy-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2-one (67 OR MPM 03 181).

[0401] Compound 14 (1g, 2.36 mmol, 1 eq.) was dissolved in tetrahydrofuran (THF) (20 mL) and cooled to 0° C. 1 M I etra-n-butylammonium fluoride in THF (9.45 mL, 9.45 mmol, 4 eq.) was added dropwise. Reaction was stirred overnight. Reaction was quenched with ammonium chloride (40 mL), extracted with ethyl acetate (3x), organic layers combined, dried with magnesium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Product was isolated as a white solid. Isolated yield: 540 milligrams, 74% Yield.

[0402] >H NMR (CD3OD, 500 MHz) 8: 7.54 (d, J = 8.0 Hz, 1H), 7.43 (d, J = 7.4 Hz, 1H), 7.24 - 7.15 (m, 3H), 6.74 (d, J = 8.8 Hz, 2H).

[0403] 13C NMR (CD3OD, 126 MHz) 8: 181.62, 158.72, 140.54 (q, J= 2.1 Hz), 136.85, 131.82, 129.90,

[0404] 128.10, 126.92 (q, J = 4.5 Hz), 125.17 (q, J= 270.5 Hz), 123.75, 116.16, 113.57 (q, J= 33.2

[0405] Hz), 77.66.

[0406] 19F NMR (CD3OD, 471 MHz) 8: -63.67.

[0407] HRMS (ESI): m / z calc, for C15H9NO3F3 [M-H]- 308.0535, found: 308.0532.

[0408] 3-(4-hydroxyphenyl)-3-(4-(prop-2-yn-l-yloxy)phenyl)-7-(trifluoromethyl)indolin-2-one (55 ORMPM 01 67). Compound 67 (1.4 g, 4.53 mmol, 1 eq.) and phenyl propargyl ether (2.32 mL, 18.1 mmol, 4 eq.) were dissolved in dichloromethane (40 mL) and cooled to 0° C. Triflic acid (1.73 mL, 22.65 mmol, 5 eq.) was added dropwise. Reaction was stirred for 30 minutes and quenched with NH4C1 (80 mL), extracted with ethyl acetate (3x), organic layers combined, dried with magnesium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Product was isolated as an off-white solid. Isolated yield: 2.04 grams, 89% Yield.

[0409] 1H NMR (CD3OD, 500 MHz) 8: 7.50 (d, J = 8.0 Hz, 1H), 7.40 (d, J = 7.5 Hz, 1H), 7.18 (t, J = 7.8 Hz, 1H), 7.13 (d, J = 8.9 Hz, 2H), 7.02 (d, J = 8.8 Hz, 2H), 6.93 (d, J = 9.0 Hz, 2H), 6.72 (d, J= 8.7 Hz, 2H), 4.70 (d, J= 2.4 Hz, 2H), 2.91 (t, J= 2.3 Hz, 1H).

[0410] 13C NMR (CD3OD, 126 MHz) 8: 182.15, 158.51, 158.13, 139.75 (q, J = 2.3 Hz), 137.74, 135.59, 133.21, 131.01, 130.52, 130.49, 125.78 (q, J = 4.5 Hz), 125.22 (q, J = 270.8 Hz), 123.37, 116.31, 115.91, 113.58 (q, J= 33.5 Hz), 79.67, 76.80, 62.14, 56.60.

[0411] 19F NMR (CD3OD, 471 MHz) 8: -63.65.

[0412] HRMS (ESI): m / z calc, for C24H17NO3F3 [M+H]+ 424.1161, found: 424.1166.

[0413] (R)-3-(4-hydroxyphenyl)-3-(4-(prop-2-yn-l-yloxy)phenyl)-7-(trifluoromethyl)indolin-2-one ((R)-55 ORMPM 02 133A). Enantiomers of 55 were separated by EDC-coupling Boc-Alanine to the phenol, separating enantiomers using ACCQ Prep HP125 equipped with Lux Cellulose-1 column using 12% isopropanol / hexanes as the mobile phase, followed by saponification of the enantiopure phenolic esters using lithium hydroxide in methanol.

[0414] 1H NMR (CD3OD, 500 MHz) 8: 7.51 (d, J= 7.7 Hz, 1H), 7.41 (d, J= 7.5 Hz, 1H), 7.18 (t, J = 7.8 Hz, 1H), 7.13 (d, J = 8.9 Hz, 2H), 7.02 (d, J = 8.8 Hz, 2H), 6.93 (d, J = 8.9 Hz, 2H), 6.73 (d,J= 8.8 Hz, 2H), 4.70 (d, J= 2.4 Hz, 2H), 2.92 (t, J= 2.4 Hz, 1H).13C NMR (CD3OD, 126 MHz) δ: 182.15, 158.51, 158.14, 139.75 (q, J = 1.9 Hz), 137.74, 135.59, 133.21, 131.01, 130.52, 130.49, 125.78 (q, J = 4.7 Hz), 125.22 (q, J = 270.7 Hz), 123.37, 116.31, 115.91, 113.58 (q, J= 33.2 Hz), 79.67, 76.80, 62.14, 56.60.

[0415] 19F NMR (CD3OD, 471 MHz) δ: -65.18.

[0416] HRMS (ESI): m / z calc. for C24H17NO3F3 [M+H]+ 424.1161, found: 424.1157.

[0417] (S)-3-( 4-hydroxyphenyl)-3-( 4-(prop-2-yn-l-yloxy)phenyl)-7-( trifluoromethyl)indolin-2-one ((S)-55 ORMPM 02 133B).

[0418] >H NMR (CD3OD, 500 MHz) 8: 7.50 (d, J = 8.0 Hz, 1H), 7.40 (d, J = 7.5 Hz, 1H), 7.18 (t, J = 7.8 Hz, 1H), 7.13 (d, J = 8.9 Hz, 2H), 7.02 (d, J = 8.8 Hz, 2H), 6.93 (d, J = 8.9 Hz, 2H), 6.72 (d, J= 8.7 Hz, 2H), 4.70 (d, J= 2.3 Hz, 2H), 2.91 (t, J= 2.4 Hz, 1H).

[0419] 13C NMR (CD3OD, 126 MHz) 8: 182.15, 158.51, 158.13, 139.75 (q, J = 2.1 Hz), 137.74, 135.59, 133.21, 131.01, 130.52, 130.49, 125.78 (q, J = 4.4 Hz), 125.22 (q, J = 271.2 Hz), 123.37, 116.31, 115.91, 113.58 (q, J= 33.2 Hz), 79.67, 76.80, 62.14, 56.60.

[0420] 19F NMR (CD3OD, 471 MHz) 8: -63.67.

[0421] 3-(4-hydroxyphenyl)-3-(4-(pent-4-yn-l-yloxy)phenyl)-7-(trifluoromethyl)indolin-2-one (56 or MPM 02 107). Compound 67 (50 mg, 0.16 mmol, 1 eq.) was dissolved in di chloromethane (2 m £ £L). BF3-OEt2 (10 uL, 0.08 mmol, 0.5 eq.) was added and the mixture was stirred for 5 minutes. (Pent-4- yn-l-yloxy)benzene (77 mgs, 0.48 mmol, 3 eq.) was added and the reaction was heated to 40° C for 6 hours. Reaction was concentrated and purified directly via flash chromatography using ethyl acetate / hexanes as the mobile phase. Product was isolated as a white solid. Isolated yield: 58 milligrams, 82% yield.

[0422] >H NMR (CD3OD, 500 MHz) 8: 7.50 (d, J = 8.0 Hz, 1H), 7.40 (d, J = 7.5 Hz, 1H), 7.17 (t, J = 7.8 1H), 7.11 (d, J = 8.9 Hz, 2H), 7.02 (d, J = 8.7 Hz, 2H), 6.86 (d, J = 8.9 Hz, 2H), 6.72 (d, J= 8.8

[0423] 2H), 4.04 (t, J = 6.1 Hz, 2H), 2.35 (td, J = 7.1, 2.6 Hz, 2H), 2.23 (t, J = 2.6 Hz, 1H), 1.93 (p, J = 6.6 Hz, 2H).

[0424] 13C NMR (CD3OD, 126 MHz) 8: 182.25, 159.81, 158.10, 139.74 (q, J = 2.1 Hz), 137.85, 134.82, 133.30, 131.00, 130.52, 125.73 (q, J = 4.6 Hz), 125.23 (q, J = 271.0 Hz), 123.34, 116.28, 116.25, 115.50, 113.54 (q, J= 33.9 Hz), 84.08, 70.01, 67.36, 62.13, 29.44, 15.71.19F NMR (CD3OD, 471 MHz) δ: -63.66.

[0425] HRMS (ESI): m / z calc, for C26H21NO3F3 [M+H]+ 452.1474, found: 452.1476.

[0426] N-(2-(2-(2-(4-((4-(3-( 4-hydroxyphenyl)-2-oxo- 7-( trifluoromethyl)indolin-3- yl)phenoxy)methyl)-lH-l, 2, 3-triazol-l-yl)ethoxy)ethoxy)ethyl)-5-( ( 3aS, 4S, 6aR) -2 -oxohexahydro- 1H- thieno[3,4-d]imidazol-4-yl)pentanamide (57 OR MPM 02 211). Compound 55 (25 mg, 0.06 mmol, 1 eq.) and Biotin- Azide (24 mg, 0.06 mmol, 1 eq.) were dissolved in ImL tBuOH / H2O (1:1). Sodium ascorbate (2 mg, 0.009 mmol, .15 eq.) was dissolved in 100 uL ofH2O and added. CUSO4 (1 mg, 0.003 mmol, .05 eq.) was dissolved in 100 uL of H2O and added and the reaction was stirred overnight. Reaction was diluted in water, extracted with ethyl acetate (3x), organic layers combined, dried with magnesium sulfate, concentrated, and purified via flash chromatography using MeOH / DCM as the mobile phase. Product was isolated as a white solid. Isolated yield: 8 milligrams, 16% yield.

[0427] 1H NMR (CD3OD, 500 MHz) 8: 8.09 (d, J= 1.4 Hz, 1H), 7.51 (d, J= 8.0 Hz, 1H), 7.41 (d, J= 7.5 Hz, 1H), 7.29 - 7.17 (m, 3H), 7.01 (d, J= 8.5 Hz, 2H), 6.97 (d, J= 8.9 Hz, 2H), 6.72 (d, J= 8.7 Hz, 2H), 5.17 (s, 2H), 4.59 (dd, J= 5.5, 4.5 Hz, 2H), 4.45 (ddd, J= 7.9, 4.9, 0.9 Hz, 1H), 4.25 (ddd, J= 8.0, 4.5, 1.1 Hz, 1H), 3.89 (dd, J= 5.5, 4.5 Hz, 2H), 3.57 (dd, J= 3.6, 1.6 Hz, 2H), 3.55 - 3.50 (m, 2H), 3.45 (t, J= 5.5 Hz, 2H), 3.31 (m, 2H), 3.20 - 3.09 (m, 1H), 2.88 (ddd, J= 12.8, 5.0, 1.3 Hz, 1H), 2.67 (d, J = 12.7 Hz, 1H), 2.17 (ddd, J = 8.2, 6.8, 1.5 Hz, 2H), 1.79 - 1.34 (m, 6H). (2 protons appear to be under Methanol solvent peak).

[0428] 19F NMR (CD3OD, 471 MHz) 8: -63.64.

[0429] HRMS (ESI): m / z calc, for C40H45N7O7SF3 [M+H]+ 824.3053, found: 824.3077.

[0430] 7-((trimethylsilyl)ethynyl)indoline-2, 3-dione (68 ORMPM 04 253). An oven dried flask was charged with 7-iodoisatin (500 mgs, 1.8 mmol, 1 eq.), tetrakis (triphenylphosphine) palladium (13 mgs, 0.04 mmol, 0.02 eq.), and copper iodide (35 mgs, 0.18 mmol, 0.1 eq.) and dimethylformamide (10 mL) was added. Tri ethylamine (631 uL, 4.5 mmol, 2.5 eq.) was added and the reaction was stirred for 1 hour. Ethynyltrimethylsilane (972 uL, 2.7 mmol, 1.5 eq.) was added and the reaction was heated to 80° C and stirred for 1 hour. Reaction was quenched with water, extracted with ethyl acetate (3x), organic layers combined, dried with magnesium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 225 milligrams, 51% yield.

[0431] 1H NMR (500 MHz, Acetone) 8: 7.66 (dd, J= 7.8, 1.2 Hz, 1H), 7.56 (dt, J= 7.5, 1.0 Hz, 1H), 7.14 (t, .J = 7,7 Hz, 1H), 0.25 (s, 9H).

[0432] 13C NMR (126 MHz, Acetone) 8: 184.14, 159.76, 152.92, 141.14, 125.55, 123.85, 119.25, 108.47, 102.06, 98.24, -0.20.

[0433] HRMS (ESI): m / z calc, for C13 H12 N 02 Si [M-H]- 242.0637, found: 242.0637.

[0434] 3-( 4-( ( tert-butyldimethylsilyl)oxy)phenyl)-3-hydroxy-7-( ( trimethylsilyl)ethynyl)indolin-2-one (69 ORMPM 04 193). In an oven-dried flask, (4-bromophenoxy)(tert-butyl)dimethyl silane (517 uL, 2.1 mmol, 2.3 eq.) was dissolved in tetrahydrofuran (10 mL) and cooled to -78° C .1.6M n- ButylLithium in hexanes (1.2 mL, 1.9 mmol, 2.1 eq.) was added dropwise and the reaction was stirred at -78° C for 30 minutes. Compound 68 (225 mgs, 0.92 mmol, 1 eq.) was added dropwise in tetrahydrofuran (2 mL) and the reaction was stirred at -78° C for 1 hour. Reaction was quenched with water, extracted with ethyl acetate (3x), organic layers combined, dried with magnesium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 200 milligrams, 48% yield.

[0435] 'H NMR (CD3OD, 500 MHz) 8: 7.32 (dd, J= 7.9, 1.3 Hz, 1H), 7.25 (d, J= 8.8 Hz, 2H), 7.18 (dd, J = 7.5, 1.2 Hz, 1H), 7.02 (t, J= 7.7 Hz, 1H), 6.80 (d, J= 8.7 Hz, 2H), 0.98 (s, 9H), 0.27 (s, 9H), 0.18 (s, 6H).

[0436] 13C NMR (CD3OD, 126 MHz) 8: 181.46, 156.93, 144.66, 134.95, 134.41, 133.81, 128.12, 126.38, 123.83, 120.95, 107.07, 100.55, 100.07, 78.96, 26.12, 19.04, -0.06, -4.36.

[0437] HRMS (ESI): m / z calc, for C25 H32 N 02 S12 [M+H]+ 452.1999, found: 434.1969 (loss of -OH).

[0438]

[0439] 7-ethynyl-3-(4-hydroxyphenyl)-3-((2-(3-(trifluoromethyl)-3H-diazirin-3- yl) ethyl) amino)indolin-2 -one (58 OR MPM 04 25). In a flame dried flask under argon, 69 (59 mg, 0.13 mmol, 1 eq.) was dissolved in tetrahydrofuran (2 mL) and cooled to 0° C. Pyridine (32 uL, 0.39 mmol, 3 eq.) was added dropwise followed by thionyl chloride (47 uL, 0.65 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0440] In a separate flame dried flask, 3 / 7-Diazirine-3 -ethanamine, 3 -(trifluoromethyl)-, hydrochloride (1:1) (50 mg, 0.26 mmol, 2 eq.) and cesium carbonate (108 mg, 0.33 mmol, 2.5 eq.) were stirred in di chloromethane (2 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (2 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred for 4 hours. Reaction was quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in methanol (2 mL) under argon and 3 M potassium hydrogen fluoride in water (108 uL, 0.33 mmol, 2.5 eq.) was added dropwise. Reaction stirred for 1 hour at room temperature, methanol removed under reduced pressure, and crude mixture purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 10 milligrams, 20% yield over 3 steps.

[0441] 'H NMR (CD3OD, 500 MHz) 8: 7.32 (dd, J= 7.8, 1.2 Hz, 1H), 7.28 (d, J= 8.8 Hz, 2H), 7.22 (dd, J = 7.5, 1.2 Hz, 1H), 7.03 (t, J= 7.7 Hz, 1H), 6.73 (d, J= 8.8 Hz, 2H), 3.77 (s, 1H), 2.30 (dt, J= 11.5, 5.9 Hz, 1H), 2.00 - 1.85 (m, 3H).

[0442] 13C NMR (CD3OD, 126 MHz) 8: 182.15, 158.53, 145.01, 133.17, 133.07, 132.19, 128.39, 126.75, 123.94 (q, J = 274.1 Hz), 123.59, 116.30, 106.19, 83.89, 79.28, 71.09, 38.36, 27.86 (q, J = 39.9 Hz), 26.69.

[0443] 19F NMR (CD3OD, 471 MHz) 8: -71.91.

[0444] HRMS (ESI): m / z calc, for C20 H14 N4 02 F3 [M-H]- 399.1069, found: 399.1061.

[0445]

[0446] 7-ethynyl-3-( 4-hydroxyphenyl)-3-( 4-( 3-( trifluoromethyl)-3H-diazirin-3-yl)piperidin-l- yl)indolin-2-one (59 OR MPM 04 139) . In a flame dried flask under argon, 69 (96 mg, 0.21 mmol, 1 eq.) was dissolved in tetrahydrofuran (6 mL) and cooled to 0° C. Pyridine (51 uL, 0.63 mmol, 3 eq.) was added dropwise followed by thionyl chloride (78 uL, 1.05 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0447] In a separate flame dried flask, Piperidine, 4-[3-(trifluoromethyl)-377-diazirin-3-yl]-, hydrochloride (1:1) (75 mg, 0.32 mmol, 1.5 eq.) and cesium carbonate (141 mg, 0.42 mmol, 2 eq.) were stirred in di chloromethane (6 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (2 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred overnight. Reaction was quenched with water, extracted with di chloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in methanol (6 mL) under argon and 3 M potassium hydrogen fluoride in water (250 uL) was added dropwise. Reaction stirred for 1 hour at room temperature, methanol removed under reduced pressure, and crude mixture purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 55 milligrams, 59% yield over 3 steps.

[0448] 'H NMR (CD3OD, 500 MHz) 8: 7.35 - 7.30 (m, 1H), 7.28 - 7.20 (m, 3H), 7.02 (m, 1H), 6.72 (d, J = 8.7 Hz, 2H), 3.76 (s, 1H), 2.81 (m, 2H), 2.27 (td, J= 11.6, 2.3 Hz, 2H), 2.15 (t, J= 4.0 Hz, 1H), 1.52 (m, 2H), 1.03 (m, 2H).

[0449] 13C NMR (CD3OD, 126 MHz) 8: 180.01, 158.66, 144.97, 132.99, 131.32, 130.24, 129.63, 127.39, 124.0 (q, J = 272.0 Hz), 123.22, 116.35, 106.10, 84.00, 79.28, 75.75, 47.76, 47.47, 33.41, 31.12 (q, J = 32.1 Hz), 28.11, 27.94.

[0450] 19F NMR (CD3OD, 471 MHz) 8: -70.54.

[0451] HRMS (ESI): m / z calc, for C23 H18 N4 02 F3 [M-H]- 439.1382, found: 439.1383.

[0452]

[0453] 7-ethynyl-3-( 4-hydroxyphenyl)-3-( 3-( 3-( trifluoromethyl)-3H-diazirin-3-yl)pyrrolidin-l- yl)indolin-2-one (60 ORMPM 04 153). In a flame dried flask under argon, 69 (72 mg, 0.16 mmol, 1 eq.) was dissolved in tetrahydrofuran (2 mL) and cooled to 0° C. Pyridine (40 uL, 0.48 mmol, 3 eq.) was added dropwise followed by thionyl chloride (58 uL, 0.8 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0454] In a separate flame dried flask, Pyrrolidine, 3-[3-(trifluoromethyl)-3 / / -diazirin-3-yl]-, hydrochloride (1:1) (50 mg, 0.23 mmol, 1.5 eq.) and cesium carbonate (101 mg, 0.31 mmol, 2 eq.) were stirred in di chloromethane (2 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (2 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred overnight. Reaction was quenched with water, extracted with di chloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated. Crude mixture was dissolved in methanol (2 mL) under argon and 3 M potassium hydrogen fluoride in water (133 uL) was added dropwise. Reaction stirred for 1 hour at room temperature, methanol removed under reduced pressure, and crude mixture purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 43 milligrams, 64% yield over 3 steps. Tested as mixture of diastereomers.

[0455] 'H NMR (CD3OD, 500 MHz) 8: 7.33 - 7.19 (m, 8H), 7.01 (td, J = 7.7, 4.6 Hz, 2H), 6.76 (d, J = 8.8 Hz, 4H), 3.76 (d, J = 1.5 Hz, 2H), 2.96 - 2.88 (m, 2H), 2.79-2.55 (m, 4H), 2.45 (m, 2H), 2.15 - 2.03 (m, 2H), 1.94 (dtd, J= 13.0, 8.6, 4.3 Hz, 2H), 1.58 (ddq, J= 34.9, 13.7, 7.2 Hz, 2H).

[0456] 19F NMR (CD3OD, 471 MHz) 8: -71.15 (d, J= 13.9 Hz).

[0457] HRMS (ESI): m / z calc, for C22 H16 N4 02 F3 [M-H]- 425.1225, found: 425.1224.

[0458]

[0459] 7-ethynyl-3-( 4-hydroxyphenyl)-3-( 3-( 3-( trifluoromethyl)-3H-diazirin-3-yl)azetidin-l- yl)indolin-2-one (61 ORMPM 04 289). In a flame dried flask under argon, 69 (113 mg, 0.25 mmol,

[0460] I eq.) was dissolved in tetrahydrofuran (5 mL) and cooled to 0° C. Pyridine (96 uL, 1.1 mmol, 4.5 eq.) was added dropwise followed by thionyl chloride (61 uL, 0.8 mmol, 3.5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0461] In a separate flame dried flask, Azetidine, 3-[3-(trifluoromethyl)-3i / -diazirin-3-yl]-, hydrochloride (1:1) (100 mg, 0.5 mmol, 2 eq.) and cesium carbonate (203 mg, 0.62 mmol, 2.5 eq.) were stirred in di chloromethane (5 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (2 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred overnight. Reaction was quenched with water, extracted with di chloromethane (3x), organic layers were combined and dried with MgSCL, filtered and concentrated. Crude mixture was dissolved in tetrahydrofuran (6 mL) under argon and 1 M tetra-butyl ammonium fluoride in THF (750 uL, 0.75 mmol, 3 eq.) was added dropwise. Reaction was quenched with sodium bicarbonate, extracted with ethyl acetate (3x), organic layers combined, dried with magnesium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield:

[0462] I I milligrams, 11% yield over 3 steps.

[0463] 'H NMR (CD3OD, 500 MHz) 8: 7.30 - 7.28 (m, 1H), 7.25 - 7.20 (m, 2H), 7.17 - 7.14 (m, 1H), 7.05 - 6.95 (m, 1H), 6.74 (d, J = 8.8 Hz, 2H), 3.77 (s, 1H), 3.53 (t, J = 7.5 Hz, 1H), 3.20 - 3.10 (m, 2H), 2.98 (t, J= 5.7 Hz, 1H), 2.75 (dd, J= 7.3, 4.9 Hz, 1H).

[0464] 13C NMR (CD3OD, 126 MHz) 8: 179.66, 158.63, 144.82, 133.26, 131.50, 129.20, 126.80, 123.84 (q, J= 275.4 Hz), 123.52, 116.44, 106.10, 83.97, 79.22, 72.55, 49.63, 29.76 (q, J= 39.4 Hz), 26.69.

[0465] 19F NMR (CD3OD, 471 MHz) 8: -71.35.

[0466] HRMS (ESI): m / z calc, for C21 H16 N4 02 F3 [M+H]+ 413.1225, found: 413.1226.

[0467]

[0468] 3-(4-((tert-butyldimethylsilyl)oxy)phenyl)-3-((2-(prop-2-yn-l-yloxy)phenyl)amino)-7- (trifluoromethyl)indolin-2-one (62a ORMPM 03 13). In a flame dried flask under argon, 14 (1g, 2.75 mmol, 1 eq.) was dissolved in tetrahydrofuran (25 mL) and cooled to 0° C. Pyridine (667 uL, 8.25 mmol, 3 eq.) was added dropwise followed by thionyl chloride (1 mL, 13.75 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0469] In a separate flame dried flask, 2-(2-propyn-l-yloxj.')-aniline (870 uL, 6.0 mmol, 2.2 eq.) and cesium carbonate (1.79 g, 5.5 mmol, 2 eq.) were stirred in dichloromethane (30 mL) for 10 minutes at 0° C. Tertiary chloride was dissolved in dichloromethane (5 mL) and added dropwise to the solution. Reaction was allowed to warm to room temperature and stirred overnight. Reaction was quenched with water, extracted with ethyl acetate (3x), organic layers were combined and dried with MgSO4, filtered and concentrated and purified via flash chromargraphy using ethyl acetate / hexanes as the mobile phase. Product was a yellow oil. Isolated yield: 720 milligrams, 55% yield over 2 steps.

[0470] 1H NMR (CD3OD, 500 MHz) 8: 7.52 (d, J= 8.1 Hz, 1H), 7.47 (d, J= 7.5 Hz, 1H), 7.39 (d, J= 8.9 Hz, 2H), 7.13 (t, J= 7.8 Hz, 1H), 6.91 (dd, J= 7.9, 1.5 Hz, 1H), 6.84 (d, J= 8.9 Hz, 2H), 6.61 (td, J= 7.7, 1.7 Hz lH), 6.55 (td, J= 7.7, 1.5 Hz, 1H), 6.04 (dd, J = 7.9, 1.6 Hz, 1H), 4.73 (d, J= 2.4 Hz, 2H), 2.93 (t, J= 2.4 Hz, 1H), 0.97 (s, 9H), 0.18 (s, 6H).

[0471] 13C NMR (CD3OD, 126 MHz) 8: 180.97, 157.56, 147.27, 140.04 (q, J = 2.2 Hz), 136.59, 134.13, 133.79, 130.01, 128.85, 126.85 (q, J = 4.5 Hz), 125.15 (q, J = 271.2 Hz), 123.68, 122.70, 121.57, 119.70, 113.89 (q, J= 32.2 Hz), 113.71, 113.39, 79.81, 77.05, 67.90, 57.47, 26.11, 19.01, -4.32.

[0472] 19F NMR (CD3OD, 471 MHz) 8: -63.36.

[0473] HRMS (ESI): m / z calc, for C30H32N2O3F3S1 [M+H]+ 553.2134, found: 553.2126.

[0474] 3-(4-amino-3-(prop-2-yn-l-yloxy)phenyl)-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2- one (62b OR MPM 03 19). The A-l inked aniline, Compound 62a (285 mgs, 0.52 mmol, 1 eq.) was dissolved in benzene (4 mL) at room temperature. Triflic acid (100 uL, 1.04 mmol, 2 eq.) was added dropwise to the solution. Reaction was stirred for 1 hour. Mixture was quenched with water (10 mL), extracted with ethyl acetate (3x), organic layers combined, dried with sodium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Product was an off- white solid. Isolated yield: 82 milligrams, 36% yield.

[0475] 1H NMR (CD3OD, 500 MHz) 8: 7.49 (d, J = 8.0 Hz, 1H), 7.41 (d, J = 7.4 Hz, 1H), 7.17 (t, J = 7.8 Hz, 1H), 7.03 (d, J = 8.8 Hz, 2H), 6.86 (d, J= 2.0 Hz, 1H), 6.72 (d, J= 8.9 Hz, 2H), 6.69 (d, J= 8.3 Hz, 1H), 6.60 (dd, J= 8.2, 2.0 Hz, 1H), 4.62 (d, J= 1.9 Hz, 2H), 2.83 (t, J= 2.3 Hz, 1H).

[0476] 13C NMR (CD3OD, 126 MHz 8 182.48, 158.02, 146.69, 139.69, 138.13, 138.02, 133.41, 132.04, 131.00, 130.59, 125.60 (q, J = 4.7 Hz), 125.30 (q, J= 270.8 Hz), 123.28, 122.83, 116.26, 114.62, 113.48 (q, J= 33.1 Hz), 79.78, 76.93, 62.37, 57.10.

[0477] 19F NMR (CD3OD, 471 MHz) 8: -63.62.

[0478] HRMS (ESI): m / z calc, for C24H18N2O3F3 [M+H]+ 439.1270, found: 439.1265.

[0479] 3-(4-azido-3-(prop-2-yn-l-yloxy)phenyl)-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2- one (62 OR MPM 03 23). The C-linked aniline, Compound 62b (130 mgs, 0.3 mmol, 1 eq.) was dissolved in ethyl acetate (4 mL) and cooled to 0° C. Concentrated 12 M hydrochloric acid (100 uL, 1.2 mmol, 4 eq.) was added dropwise. Sodium nitrite (31 mgs, 0.45 mmol, 1.5 eq.) was dissolved in water (100 uL) and added dropwise. Reaction mixture was stirred for 1 hour at 0° C. Sodium azide (30 mgs, 0.45 mmol, 1.5 eq.) was dissolved in water (100 uL) and added dropwise, reaction was allowed to warm to room temperature over 1 hour. Reaction was quenched with water (10 mL), extracted with Ethyl Acetate (3x), organic layers combined, dried with sodium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Product was isolated as an off- white solid. Isolated yield: 78 milligrams, 57% yield.

[0480] 1H NMR (CD30D, 500 MHz) 8: 7.52 (d, J= 8.1 Hz, 1H), 7.45 (d, J= 7.5 Hz, 1H), 7.20 (t, J = 7.8 Hz, 1H), 7.08 - 6.99 (m, 3H), 6.95 (d, J= 8.3 Hz, 1H), 6.82 (dd, J= 8.4, 2.0 Hz, 1H), 6.74 (d, J= 8.8 Hz, 2H), 4.68 (d, J= 1.3 Hz, 1H), 2.90 (t, J= 2.3 Hz, 1H).

[0481] 13C NMR (CD3OD, 126 MHz) 8: 181.53, 158.27, 151.39, 140.45, 139.80 (q, J = 2.1 Hz), 137.15, 132.72, 131.05, 130.55, 129.85, 126.00 (q, J = 4.7 Hz), 125.17 (q, J = 272.2 Hz), 123.49, 123.22, 121.75, 116.54, 116.45, 113.70 (q, J = 32.7 Hz), 78.87, 77.74, 62.43, 57.71.

[0482] 19F NMR (CD3OD, 471 MHz) 8: -63.68.

[0483] HRMS (ESI): m / z calc, for C24H16N4O3F3 [M+H]+ 465.1175, found: 465.1166.

[0484] IR (cm1): 3288, 2928, 2123 / 2095 (azide), 1716, 1611, 1503, 1458, 1416, 1374, 1336, 1315, 1236, 1168, 1121, 1101, 1067.

[0485] 3-(4-((tert-butyldimethylsilyl)oxy)phenyl)-3-((2-ethynylphenyl)amino)-7- (trifluoromethyl)indolin-2-one (63a ORMPM 03 165). In a flame dried flask under argon, 14 (5.4 g, 12.9 mmol, 1 eq.) was dissolved in tetrahydrofuran (60 mL) and cooled to 0° C. Pyridine (3.1 mL, 38.7 mmol, 3 eq.) was added dropwise followed by thionyl chloride (4.7 mL, 64.3 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with MgSO4, filtered and concentrated to afford the tertiary chloride (unstable).

[0486] 2-ethynyl-aniline (4.4 mL, 38.7 mmol, 3 eq.) and cesium carbonate (11.8 g, 36.1 mmol, 2.8 eq.) are dissolved in dichloromethane (50 mL) and cooled to 0° C. After being stirred for 15 minutes, the tertiary chloride was dissolved in di chloromethane (10 mL) and added dropwise. The reaction was allowed to warm up to room temperature and stirred for 16 hours. Reaction was quenched with water (100 mL), extracted with ethyl acetate (3x), organic layers combined and dried with magnesium sulfate, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Product was a yellow solid. Isolated yield: 3.16 grams, 47% yield over 2 steps.

[0487] >H NMR (CD3OD, 500 MHz) 8: 7.60 (d, J= 8.0 Hz, 1H), 7.54 (d, J= 7.5 Hz, 1H), 7.38 (d, J= 8.8 Hz, 2H), 7.29 (d, J= 7.7 Hz, 1H), 7.23 (t, J= 7.7 Hz, 1H), 6.92 (t, J= 7.8 Hz, 1H), 6.87 (d, J= 8.8 Hz, 2H), 6.62 (t, J= 7.8 Hz, 1H), 5.99 (d, J= 8.4 Hz, 1H), 3.94 (s, 1H), 0.98 (s, 9H), 0.20 (s, 6H).

[0488] 13C NMR (CD3OD, 126 MHz) δ: 180.36, 157.80, 147.83, 140.09 (q, J = 2.0 Hz), 133.63, 133.60, 133.50, 130.76, 129.99, 128.76, 127.17 (q, J = 4.5 Hz), 125.12 (q, J = 271.9 Hz) 123.90, 121.72, 119.16, 114.14 (q, J = 33.4 Hz), 112.41, 109.82, 85.41, 81.13, 67.72, 26.08, 19.04, -4.37.

[0489] 19F NMR (CD3OD, 471 MHz) δ: -62.83.

[0490] HRMS (ESI): m / z calc, for C29H29N2O2F3NaSi [M+Na]+ 545.1848, found: 545.1835.

[0491] 3-((2-ethynylphenyl)amino)-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2-one (63bl OR MPM 03 153). The TBS-protected N-linked aniline 63a (3.16 g, 6.05 mmol, 1 eq.) was dissolved in tetrahydrofuran (60 mL) and the solution cooled to 0° C. 1 M tetra-butyl ammonium fluoride in tetrahydrofuran (18.1 mL, 18.1 mmol, 3 eq.) was added dropwise over 5 minutes. Reaction was stirred for 1 hour at 0° C. Reaction was quenched with ammonium chloride (100 mL), extracted with ethyl acetate (3x), organic layers combined and dried with magnesium sulfate, concentrated and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Product was a yellow oil. Isolated yield: 2.21 grams, 89% yield.

[0492] >H NMR (CD3OD, 500 MHz) 8: 7.59 (d, J= 8.0 Hz, 1H), 7.52 (d, J= 7.4 Hz, 1H), 7.31 (d, J= 8.8 Hz, 2H), 7.29 (dd, J= 7.8, 1.1 Hz, 1H) 7.22 (t, J = 7.8 Hz, 1H), 6.93 (td, J= 8.7, 1.6 Hz, 1H), 6.79 (d, J = 8.8 Hz, 2H), 6.61 (t, J= 7.5 Hz, 1H), 5.97 (d, J= 8.3 Hz, 1H), 3.92 (s, 1H).

[0493] 13C NMR (CD3OD, 126 MHz) 8: 180.58, 159.53, 147.87, 140.06 (q, J = 2.0 Hz), 133.73, 133.57, 131.16, 130.78, 129.96, 128.73, 127.05 (q, J = 4.5 Hz), 125.14 (q, J = 271.2 Hz), 123.80, 119.08, 116.89, 114.08 q, J= 33.0 Hz), 112.31, 109.70, 85.28, 81.14, 67.63.

[0494] 19F NMR (CD3OD, 471 MHz) 8: -62.83.

[0495] HRMS (ESI): m / z calc, for C23H16N2O2F3 [M+H]+ 409.1164, found: 409.1148. 3-(4-amino-3-(prop-2-yn-l-yloxy)phenyl)-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2- one (63bII OR MPM 03 85). The deprotected N-linked aniline 63bl (2.27 g, 5.6 mmol, 1 eq.) was dissolved in benzene (50 rnL). Triflic acid (1 mL, 11.2 mmol, 2 eq.) was added dropwise. A black insoluble solid formed in the reaction vessel. Reaction allowed to proceed for 1 hour. Reaction mixture quenched with sodium bicarbonate (100 mL), extracted with ethyl acetate (3x), organic layers combined, dried with sodium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as mobile phase. Product was an off-white solid. Isolated yield: 538 milligrams, 24% yield.

[0496] 1H NMR (CD3OD, 500 MHz) 8: 7.50 (d, J = 8.2 Hz, 1H), 7.38 (d, J = 7.5 Hz, 1H), 7.18 (t, J = 7.8 Hz, 1H), 7.01 (m, 3H), 6.96 (dd, J = 8.6, 2.3 Hz, 1H), 6.73 (d, J = 8.7 Hz, 2H), 6.69 (d, J = 8.7 Hz, 1H), 3.70 (s, 1H).

[0497] 13C NMR (CD3OD, 126 MHz) 8: 182.24, 158.09, 150.29, 139.70, 137.74, 133.18, 132.96. 131.19, 130.90, 130.46, 130.40, 125.74 (q, J = 4.7Hz), 125.23 (q, J = 270.5 Hz), 123.35, 116.32, 115.53, 113.57 (q, J = 30.4 Hz), 107.48, 83.79, 81.27, 61.83.

[0498] 19F NMR (CD3OD, 471 MHz) 8: -63.65.

[0499] HRMS (ESI): m / z calc, for C23H16N2O2F3 [M+H]+ 409.1164, found: 409.1150.

[0500] 3-(4-azido-3-ethynylphenyl)-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2-one (63 OR MPM 03 103). The C-linked aniline 63bII (538 mgs, 1.3 mmol, 1 eq.) was dissolved in a 1: 1 mixture of of acetonitrile and dimethylformamide (12 mL) and cooled to 0° C. 10% sulfuric acid (2 mL) was added drop wise. A solution of sodium nitrite (91 mgs, 1.3 mmol, 1 eq.) in water (100 uL) was added dropwise, solution changed color from yellow to red. Reaction was stirred at 0° C for 30 minutes. A solution of sodium azide (85 mgs, 1.3 mmol, eq.) in water (100 uL) was added dropwise leading to gas evolution in the reaction. Reaction was quenched with water (20 mL), extracted with ethyl acetate (3x), organic layers combined, dried with sodium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Product was an off white solid. Isolated yield: 227 milligrams, 40% yield.

[0501] 1H NMR (CD3OD, 500 MHz) 8: 7.54 (d, J = 8.0 Hz, 1H), 7.44 (d, J = 7.6 Hz, 1H), 7.34 - 7.24 (m, 2H), 7.22 (t, J = 8.0 Hz, 1H), 7.19 (d, J = 8.4 Hz, 1H), 7.02 (d, J = 8.8 Hz, 2H), 6.75 (d, J = 8.8 Hz, 2H), 3.75 (s, 1H).13C NMR (CD3OD, 126 MHz) δ: 181.21, 158.38, 142.30, 139.82 (q, J = 2.1 Hz), 139.68, 136.69,

[0502] 134.84, 132.50, 131.39, 130.99, 130.43, 126.18 (q, J = 4.7 Hz), 125.13 (q, J = 271.4 Hz), 123.58,

[0503] 120.29, 116.53, 115.72, 113.81 (q, J = 33.3 Hz), 85.13, 79.63, 61.93.

[0504] 19F NMR (CD3OD, 471 MHz) δ: -62.99.

[0505] HRMS (ESI): m / z calc, for C23H14N4O2F3 [M+H]+ 435.1069, found: 435.1068.

[0506] IR (cmx): 3287. 2121 (azide). 1720, 1611, 1511,1487,1458, 1336, 1310, 1170, 1116.

[0507] (R)-3-(4-azido-3-ethynylphenyl)-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2-one ((R)- 63 orMPM 03 103A). 63 was separated into its respective enantiomers using preparative chiral HPLC separation (Lux® 5 uM Cellulose-1, 250 x 21.2 mm, AXIA™ Packed, isocratic: 10% i- PrOH / Hexanes). (R)-63 and fS')-63 isolated as off-white solids. Stereochemistry inferred from activity.1H NMR (CD3OD, 500 MHz) 8: 7.54 (d, J = 8.0 Hz, 1H), 7.44 (d, J = 7.6 Hz, 1H), 7.40 - 7.24 (m, 2H), 7.21 (t, J= 7.9 Hz, 1H), 7.18 (d, J= 8.4 Hz, 1H), 7.02 (d, J= 8.8 Hz, 2H), 6.75 (d, J= 8.7 Hz, 2H), 3.75 (s, 1H).

[0508] 13C NMR (CD3OD, 126 MHz) 8: 181.21, 158.40, 142.30, 139.82 (q, J = 2.1 Hz), 139.70, 136.70,

[0509] 134.84, 132.49, 131.39, 131.00, 130.43, 126.18 (q, J = 4.8 Hz), 125.14 (q, J = 271.4 Hz), 123.58,

[0510] 120.29, 116.53, 115.73, 113.81 (q, J = 33.3 Hz), 85.13, 79.63, 61.93.

[0511] 19F NMR (CD3OD, 471 MHz) 8: -62.99.

[0512] HRMS (ESI): m / z calc, for C23H14N4O2F3 [M+H]+ 435.1069, found: 435.1068.

[0513] (S)-3-(4-azido-3-ethynylphenyl)-3-(4-hydroxyphenyl)-7-(trifluoromethyl)indolin-2-one ((S)-63 orMPM 03 103B).1H NMR (CD3OD, 500 MHz) 8: 7.54 (d, J = 8.0 Hz, 1H), 7.44 (d, J = 7.5 Hz, 1H), 7.34 - 7.23 (m, 2H), 7.21 (t, J = 7.8 Hz, 1H), 7.18 (d, J = 8.5 Hz, 1H), 7.02 (d, J = 8.8 Hz, 2H), 6.75 (d, J = 8.7 Hz, 2H), 3.75 (s, 1H).13C NMR (CD3OD, 126 MHz) 8: 181.21, 158.41, 142.30, 139.84 (q, J = 2.1 Hz), 139.70, 136.70,

[0514] 134.84, 132.49, 131.39, 131.00, 130.43, 126.18 (q, J = 4.3 Hz), 125.12 (q, J = 271.3 Hz), 123.58,

[0515] 120.30, 116.53, 115.73, 113.81 (q, J = 32.2 Hz), 85.13, 79.63, 61.93.19F NMR (CD3OD, 471 MHz) 8: -62.99.

[0516] HRMS (ESI): m / z calc, for C23H14N4O2F3 [M+H]+ 435.1069, found: 435.1061.

[0517]

[0518] 3-( 4-( ( tert-butyldimethylsilyl)oxy)phenyl)-3-(phenylamino)-7-( ( trimethylsilyl) ethynyl)indolin- 2-one (64a OR MPM 04 107). In a flame dried flask under argon, 14 (170 mgs, 0.38 mmol, 1 eq.) was dissolved in tetrahydrofuran (4 mL) and cooled to 0° C. Pyridine (92 uL, 1.1 mmol, 3 eq.) was added dropwise followed by thionyl chloride (137 uL, 1.9 mmol, 5 eq.). Reaction was stirred until starting material was consumed by TLC and quenched with water, extracted with dichloromethane (3x), organic layers were combined and dried with magnesium sulfate, filtered and concentrated to afford the tertiary chloride (unstable).

[0519] Aniline (104 uL, 1.14 mmol, 3 eq.) and cesium carbonate (346 mg, 1.06 mmol, 2.8 eq.) are dissolved in di chloromethane (4 mL) and cooled to 0° C. After being stirred for 15 minutes, the tertiary chloride was dissolved in dichloromethane (2 mL) and added dropwise. The reaction was allowed to warm up to room temperature and stirred for 16 hours. Reaction was quenched with water, extracted with ethyl acetate (3x), organic layers combined and dried with magnesium sulfate, filtered and concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 105 milligrams, 53% yield over 2 steps.

[0520] 'H NMR (CD3OD, 500 MHz) 8: 7.38 (d, J= 8.8 Hz, 2H), 7.33 (dd, J= 7.9, 1.2 Hz, 1H), 7.26 (dd, J = 7.5, 1.2 Hz, 1H), 7.01 (t, J = 7.7 Hz, 1H), 6.98 - 6.94 (m, 2H), 6.83 (d, J = 8.8 Hz, 2H), 6.64 - 6.57 (m, 1H), 6.42 (d, J= 7.9 Hz, 1H), 0.98 (s, 9H), 0.28 (s, 9H), 0.19 (s, 6H).

[0521] 13C NMR (CD3OD, 126 MHz) 8: 181.50, 157.32, 147.33, 144.24, 134.29, 133.45, 132.91, 129.72, 129.18, 126.56, 123.66, 121.28, 119.51, 116.10, 107.31, 100.52, 100.36, 69.40, 26.11, 19.05, -0.07, - 4.37.

[0522] HRMS (ESI): m / z calc, for C31 H37 N2 02 S12 [M-H]- 525.2394, found: 525.2412.

[0523]

[0524] 3-(4-aminophenyl)-7-ethynyl-3-(4-hydroxyphenyl)indolin-2-one (64b ORMPM 04 111). 64a (65 mgs, 0.21 mmol, 1 eq.) was dissolved in methanol (2 mL) at room temperature and 3M potassium hydrogen difluoride in water (174 uL, 0.52 mmol, 2.5 eq.) was added dropwise. Reaction stirred for 1 hour at room temperature, methanol removed under reduced pressure, and crude mixture purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. The deprotected N-linked aniline (65 mgs, 0.19mmol, 1 eq.) was dissolved in benzene (3 mL) at room temperature. Trifluoroacetic acid (44 uL, 0.57 mmol, 3 eq.) was added dropwise and the reaction was heated at 45° C until completion. Reaction mixture quenched with sodium bicarbonate (100 mL), extracted with ethyl acetate (3x), organic layers combined, dried with sodium sulfate, concentrated, and purified via flash chromatography using methanol / di chloromethane as a mobile phase. Isolated yield: 54 milligrams, 79% yield over 2 steps.

[0525] 'H NMR (CD3OD, 500 MHz) 8: 7.29 (dd, J= 7.9, 1.2 Hz, 1H), 7.13 (dd, J= 7.6, 1.2 Hz, 1H), 7.04 - 6.97 (m, 3H), 6.93 (d, J= 8.7 Hz, 2H), 6.70 (d, J= 8.8 Hz, 2H), 6.64 (d, J= 8.7 Hz, 2H), 3.76 (s, 1H).13C NMR (CD3OD, 126 MHz) 8: 180.40, 155.68, 145.96, 142.41, 134.34, 131.75, 130.01, 128.39,

[0526] 127.97, 125.42, 121.13, 114.18, 113.95, 103.76, 81.54, 77.43, 61.29.

[0527] HRMS (ESI): m / z calc, for C22 H17 N2 02 [M+H]+ 341.1290, found: 341.1287.

[0528] 3-(4-azidophenyl)-7-ethynyl-3-(4-hydroxyphenyl)indolin-2-one (64 OR MPM 04 207). Compound 64b (95 mgs, 0.3, 1 eq.) was dissolved in a 1:1 mixture of acetonitrile and dimethylformamide (3 mL) and cooled to 0° C. 10% sulfuric acid (150 uL) was added dropwise. A solution of sodium nitrite (23 mgs, 0.34 mmol, 1.2 eq.) in water (50 uL) was added dropwise. Reaction was stirred at 0° C for 30 minutes. A solution of sodium azide (23 mgs, 0.34mmol, 1.2 eq.) in water (50 uL) was added dropwise leading to gas evolution in the reaction. Reaction was quenched with water, extracted with ethyl acetate (3x), organic layers combined, dried with sodium sulfate, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 63 milligrams, 62% yield.

[0529] 'H NMR (CD3OD, 500 MHz) 8: 7.34 - 7.30 (m, 1H), 7.23 (d, J = 8.7 Hz, 2H), 7.20 - 7.15 (m, 1H), 7.05 - 6.99 (m, 5H), 6.72 (d, J= 8.8 Hz, 2H), 3.78 (s, 1H).

[0530] 13C NMR (CD3OD, 126 MHz) 8: 181.56, 158.13, 144.65, 140.68, 140.10, 135.49, 133.18, 132.56,

[0531] 130.99, 130.50, 127.58, 123.48, 120.03, 116.30, 106.19, 83.91, 79.40, 63.51.

[0532] HRMS (ESI): m / z calc, for C22 H15 N4 02 [M+H]+ 367.1195, found: 367.1202.

[0533] (S)-3-(4-azidophenyl)-7-ethynyl-3-(4-hydroxyphenyl)indolin-2-one ((R)-64 OR

[0534] MPM 04 113A). 64 was separated into its respective enantiomers using preparative chiral HPLC separation (Lux® 5 uM Cellulose-1, 250 x 21.2 mm, AXIA™ Packed, isocratic: 11% i- PrOH / Hexanes). 45 milligrams of 64 dissolved in isopropanol / hexanes (1:1), filtered, and injected. (R)-64 and (8)-64 isolated as white solids. Stereochemistry inferred from activity.

[0535] 'H NMR (CD3OD, 500 MHz) 8: 7.32 (dd, J= 7.9, 1.2 Hz, 1H), 7.23 (d, J= 8.7 Hz, 2H), 7.18 (dd, J = 7.5, 1.2 Hz, 1H), 7.07 - 6.97 (m, 5H), 6.72 (d, J= 8.8 Hz, 2H), 3.78 (s, 1H).

[0536] 13C NMR (CD3OD, 126 MHz) 8: 181.56, 158.12, 144.64, 140.68, 140.09, 135.49, 133.19, 132.56,

[0537] 130.99, 130.50, 127.59, 123.49, 120.03, 116.31, 106.19, 83.92, 79.41, 63.51.

[0538] HRMS (ESI): m / z calc, for C22 H15 N4 02 [M+H]+ 367.1195, found: 367.1195.

[0539] (R)-3-(4-azidophenyl)-7-ethynyl-3-(4-hydroxyphenyl)indolin-2-one ((S)-64 OR

[0540] MPM 04 113B).

[0541] 1H NMR (CD30D, 500 MHz) 8: 7.33 (dd, J= 7.8, 1.2 Hz, 1H), 7.23 (d, J= 8.7 Hz, 2H), 7.19 - 7.16 (m, 1H), 7.07 - 6.98 (m, 5H), 6.72 (d, J= 8.9 Hz, 2H), 3.79 (s, 1H).

[0542] 13C NMR (CD3OD, 126 MHz) 8: 181.56, 158.13, 144.65, 140.68, 140.09, 135.49, 133.19, 132.56,

[0543] 130.99, 130.50, 127.59, 123.49, 120.03, 116.31, 106.19, 83.92, 79.41, 63.52.

[0544] HRMS (ESI): m / z calc, for C22 H15 N4 02 [M+H]+ 367.1195, found: 367.1198.

[0545] N-(2-(2-(2-(4-((2-azido-5-(3-(4-hydroxyphenyl) -2-oxo- 7-(trifluoromethyl)indolin-3- yl)phenoxy)methyl)-lH-l, 2, 3-triazol-l-yl)ethoxy)ethoxy)ethyl)-5-( ( 3aS, 4S, 6aR) -2 -oxohexahydro- 1H- thieno[3,4-d]imidazol-4-yl)pentanamide (65 OR MPM 03 115). The C-linked aniline 62B (0.13 mmol, 1 eq.) and Biotin-PEG3 -Azide (Lumiprobe, 50 mgs, 0.13 mmol, 1 eq.) were dissolved in 2 rnL water / tert-butanol (1:1). Sodium ascorbate (2 mgs, 0.0065 mmol, 0.05 eq.) was dissolved in water (100 uL) and added. Copper sulfate (2 mgs, 0.0065 mmol, 0.05 equiv) was dissolved in water (100 uL) and added. Reaction was stirred overnight. Reaction was quenched with water (5 mL), extracted with ethyl acetate (3x), organic layers combined and dried, and reaction was purified using normal phase chromatography with MeOH / DCM as the mobile phase.

[0546] Product was taken forward without full characterization. The triazole product (51 mgs, 0.06 mmol, 1 eq.) was dissolved in 3 mL acetonitrile / dimethylformamide (2:1) and 667 uL of 10% sulfuric acid, and cooled to 0° C. Sodium nitrite (5 mgs, 0.06 mmol, 1 eq.) was dissolved in 100 uL of water and added dropwise. Reaction was stirred for 30 minutes and then sodium azide (5 mgs, 0.06 mmol, 1 eq.) was dissolved in 100 uL of water and added dropwise. Reaction was warmed to room temperature and stirred for 30 minutes. Reaction quenched with water (6 mL), extracted with ethyl acetate (3x), organic layers combined, dried with magnesium sulfate, concentrated, and purified via flash chromatography using MeOH / DCM as the mobile phase. Product was isolated as a white solid. Isolated yield: 9 milligrams, 18% yield.

[0547] 1H NMR (CD3OD, 500 MHz) 8: 7.98 (s, 1H), 7.53 (d, J= 8.0 Hz, 1H), 7.46 (d, J= 7.5 Hz, 1H), 7.21 (t, J= 7.8 Hz, 1H), 7.04 (s, 1H), 7.01 (d, J = 8.9 Hz, 2H), 6.94 (d, J= 8.3 Hz, 1H), 6.83 - 6.76 (m, 1H), 6.75 (d, J= 8.8 Hz, 2H), 5.15 (s, 2H), 4.58 (t, J= 5.1 Hz, 2H), 4.46 (dd, J= 7.9, 5.0 Hz, 1H), 4.27 (dd, J= 7.9, 4.5 Hz, 1H), 3.88 (t, J= 5.1 Hz, 2H), 3.64 - 3.53 (m, 2H), 3.55 - 3.49 (m, 2H), 3.46 (t, J= 5.5 Hz, 2H), 3.17 - 3.07 (m, 1H), 2.88 (dd, J= 12.8, 5.0 Hz, 1H), 2.68 (d, J= 12.7 Hz, 1H), 2.18 (t, J= 7.4 Hz, 2H), 1.79 - 1.16 (m, 8H).

[0548] 19F NMR (CD3OD, 471 MHz) 8: -63.61.

[0549] HRMS (ESI): m / z calc, for C40H44N1007SF3 [M+H]+ 865.3067, found: 865.3055.

[0550] IR ^m1): 3241, 2926, 2419, 2119 (azide), 1689, 1642, 1611, 1504, 1457, 1336, 1316, 1235, 1169, 1103, 1017.

[0551] 3-(2-(3-(but-3-yn-l-yl)-3H-diazirin-3-yl)ethyl)-3-(4-((tert-butyldimethylsilyl)oxy)phenyl)-7- (trifluoromethyl)indolin-2-one (66a ORMPM 06 15). In an oven dried flask, compound 50 (53 mgs, 0.13 mmol, 1 eq.) was dissolved in dimethylformamide (2 mL) and the reaction was cooled to -40° C. Potassium tert-butoxide (16 mgs, 0.14 mmol, 1.1 eq.) was added and the reaction was allowed to warm to -14° C (yellow to red color transition) over the course of 5 minutes. Reaction was cooled to -50° C and 3H-Diazirine, 3-(3~butyn-1 -yl)-3-(2-iodoethy)- (65 mgs, 0.26 mmol, 2 eq) was added dropwise in dimethylformamide (1 mL). Reaction was allowed to warm to room temperature over the course of 5 hours. Reaction quenched with ammonium chloride, extracted with ethyl acetate (3x), the organic layers were combined, dried with MgSO4, concentrated, and purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 32 milligrams, 46% yield.

[0552] 'H NMR (CD3OD, 500 MHz) 8: 7.86 (s, 1H), 7.48 (d, J= 8.0 Hz, 1H), 7.30 (d, J= 7.4 Hz, 1H), 7.21 - 7.16 (m, 1H), 7.13 (d, J= 8.7 Hz, 2H), 6.76 (d, J= 8.8 Hz, 2H), 2.23 (td, J= 13.1, 4.7 Hz, 1H), 2.00 - 1.86 (m, 4H), 1.63 - 1.51 (m, 2H), 1.40 (ddd, J= 14.5, 13.1, 4.7 Hz, 1H), 1.07 - 0.98 (m, 1H), 0.96 (s, 9H), 0.17 (d, J= 1.9 Hz, 6H).

[0553] 13C NMR (CD3OD, 126 MHz) 8: 179.14, 155.54, 138.19, 133.86, 130.90, 128.49, 127.82, 125.41 (q, J = 4.2 Hz), 123.96 (q, J = 269.7 Hz), 120.41, 112.51 (q, J = 33.3 Hz), 82.67, 69.40, 54.77, 32.10, 32.01, 28.27, 27.96, 25.77, 18.31, 13.38, -4.28.19F NMR (CD3OD, 471 MHz) 8: -60.64.

[0554] HRMS (ESI): m / z calc, for C28 H31 N3 02 F3 Si [M-H]- 526.2138, found: 526.2134.

[0555] 3-(2-(3-(but-3-yn-l-yl)-3H-diazirin-3-yl)ethyl)-3-(4-hydroxyphenyl)-7- (trifluoromethyl)indolin-2-one (66 OR MPM 06 17). Compound 66 (53 mgs, 0.1 mmol, 1 eq.) was dissolved in methanol (5 mL) at room temperature. 3 M potassium hydrogen fluoride in water (83 uL, 0.25 mmol, 2.5 eq.) was added dropwise. Upon disappearance of starting material by TLC, methanol was removed under reduced pressure and the crude mixture was purified via flash chromatography using ethyl acetate / hexanes as the mobile phase. Isolated yield: 33 milligrams, 80% yield.

[0556] 'H NMR (CD3OD, 500 MHz) 8: 7.83 (s, 1H), 7.48 (d, J= 8.0 Hz, 1H), 7.29 (d, J= 7.4 Hz, 1H), 7.19 (t, J= 7.7 Hz, 1H), 7.12 (d, J = 8.6 Hz, 2H), 6.73 (d, J = 8.7 Hz, 2H), 5.18 (s, 1H), 2.23 (td, J= 13.2, 4.7 Hz, 1H), 2.00 - 1.88 (m, 4H), 1.59 - 1.53 (m, 2H), 1.44 - 1.36 (m, 1H), 1.00 (ddd, J= 14.0, 12.7, 4.1 Hz, 1H).

[0557] 13C NMR (CD3OD, 126 MHz) 8: 179.33, 155.50, 138.10 (q, J = 2.2 Hz), 133.90, 130.35, 128.44, 128.09, 125.49 (q, J = 4.2 Hz), 123.94 (q, J = 272.2 Hz), 122.91, 115.89, 112.61 (q, J = 33.1 Hz), 82.68, 69.43, 54.79, 32.08, 31.74, 28.24, 27.95, 13.37.19F NMR (CD3OD, 471 MHz) 8: -60.64.

[0558] HRMS (ESI): m / z calc, for C22 H17 N3 02 F3 [M-H]- 412.1273, found: 412.1266.

[0559] (S)-3-(2-(3-(but-3-yn-l-yl)-3H-diazirin-3-yl)ethyl)-3-(4-hydroxyphenyl)-7-(trifluoro- methyl)indolin-2-one ((S)-66 OR MPM 06 _17A). 66 was separated into its respective enantiomers using preparative chiral HPLC separation (Lux® 5 uM Cellulose-1, 250 x 21.2 mm, AXIA™ Packed, isocratic: 7.5% i- PrOHHexanes). 33 milligrams of 66 dissolved in isopropanol / hexanes (1:1), filtered, and injected. (R)-66 and (S)-66 isolated as white solids. Stereochemistry inferred from activity.

[0560] 'H NMR (CD3OD, 500 MHz) 8: 7.83 (s, 1H), 7.48 (d, J= 8.0 Hz, 1H), 7.29 (d, J= 7.4 Hz, 1H), 7.20 (m, 1H), 7.12 (d, J= 8.7 Hz, 2H), 6.74 (d, J= 8.7 Hz, 2H), 5.18 (s, 1H), 2.23 (td, J= 13.1, 4.8 Hz, 1H), 2.00 - 1.87 (m, 4H), 1.61 - 1.53 (m, 2H), 1.40 (ddd, J= 14.3, 12.9, 4.7 Hz, 1H), 1.01 (ddd, J = 14.3, 12.8, 4.1 Hz, 1H).

[0561] 13C NMR (CD3OD, 126 MHz) 8: 179.31, 155.48, 138.09, 133.89, 130.38, 128.44, 128.09, 125.49 (q, J = 4.0 Hz), 123.94 (q, J = 271.9 Hz), 122.90, 115.89, 112.60 (q, J = 33.2 Hz), 82.68, 69.43, 54.78, 32.09, 31.77, 28.24, 27.96, 13.38.19F NMR (CD3OD, 471 MHz) 8: -60.62.

[0562] HRMS (ESI): m / z calc, for C22 H17 N3 02 F3 [M-H]- 412.1273, found: 412.1271.

[0563] (R)-3-(2-(3-(but-3-yn-l-yl)-3H-diazirin-3-yl)ethyl)-3-(4-hydroxyphenyl)-7- (trifluoromethyl)indolin-2-one ((R)-66 OR MPM 06 _17B).

[0564] 'H NMR (CD3OD, 500 MHz) 8: 7.75 (s, 1H), 7.48 (d, J= 8.0 Hz, 1H), 7.29 (d, J= 7.4 Hz, 1H), 7.19 (t, .7 = 7,8 Hz, 1H), 7.13 (d, J= 8.6 Hz, 2H), 6.75 (d, J= 8.8 Hz, 2H), 5.03 (s, 1H), 2.23 (td, J= 13.1, 4.7 Hz, 1H), 2.00 - 1.89 (m, 4H), 1.57 - 1.52 (m, 2H), 1.47 - 1.36 (m, 1H), 1.01 (ddd, J= 14.0, 12.7, 4.1 Hz, 1H).

[0565] 13C NMR (CD3OD, 126 MHz) 8: 179.12, 155.44, 138.11, 133.84, 130.47, 128.45, 128.11, 125.49 (q, J = 4.1 Hz), 123.95 (q, J = 275.2 Hz), 122.87, 115.87, 112.57 (q, J = 33.3 Hz), 82.67, 69.43, 54.74, 32.10, 31.85, 28.25, 27.95, 13.38.19F NMR (CD3OD, 471 MHz) 8: -60.62.

[0566] HRMS (ESI): m / z calc, for C22 H17 N3 02 F3 [M-H]- 412.1273, found: 412.1266.

[0567] Example 3. Toxicity assessment.

[0568] Table 16. Ames Mutagenicity of ErSO, ErSO-DFP, and ErSO-TFPy.

[0569] 96-well setup ofMuta-ChromePlate Basic Kit Version 2.2. Quantified revertant colonies across TAI 00, TA98, and TA97a when treated with 50 pM compound (n = 3). Compound incubated with bacteria for 5 days except for TA97a (* = 4 days).

[0570] Table 17. Maximum tolerated dose assessment.

[0571] Example 4. Pharmaceutical Dosage Forms.

[0572] The following formulations illustrate representative pharmaceutical dosage forms that may be used for the therapeutic or prophylactic administration of a compound of a formula described herein, a compound specifically disclosed herein, or a pharmaceutically acceptable salt or solvate thereof (hereinafter referred to as 'Compound X'):

[0573] (i) Tablet 1 mg / tablet

[0574] 'Compound X' 100.0

[0575] Lactose 77.5

[0576] Povidone 15.0

[0577] Croscarmellose sodium 12.0

[0578] Microcrystalline cellulose 92.5

[0579] Magnesium stearate 3,0

[0580] 300.0

[0581] (ii) Tablet 2 mg / tablet

[0582] 'Compound X' 20.0

[0583] Microcrystalline cellulose 410.0

[0584] Starch 50.0

[0585] Sodium starch glycolate 15.0 Magnesium stearate 5,0

[0586] 500.0

[0587] (iii) Capsule mg / capsule 'Compound X' 10.0 Colloidal silicon dioxide 1.5 Lactose 465.5

[0588] Pregelatinized starch 120.0 Magnesium stearate 3,0

[0589] 600.0

[0590] (iv) Injection 1 (1 mg / mL) mg / mL 'Compound X' (free acid form) 1.0 Dibasic sodium phosphate 12.0 Monobasic sodium phosphate 0.7 Sodium chloride 4.5

[0591] LO N Sodium hydroxide solution q.s. (pH adjustment to 7.0-7.5) Water for injection q.s. ad 1 mL

[0592] (v) Injection 2 (10 mg / mL) mg / mL 'Compound X' (free acid form) 10.0 Monobasic sodium phosphate 0.3 Dibasic sodium phosphate 1.1 Polyethylene glycol 400 200.0

[0593] 0.1 N Sodium hydroxide solution q.s. (pH adjustment to 7.0-7.5) Water for injection q.s. ad 1 mL

[0594] (vi) Aerosol mg / can 'Compound X' 20 Oleic acid 10

[0595] T ri chloromonofluoromethane 5,000 Dichlorodifluoromethane 10,000 Dichlorotetrafluoroethane 5,000

[0596] (vii) Topical Gel 1 wt.% 'Compound X' 5% Carbomer 934 1.25% Triethanolamine q.s.

[0597] (pH adjustment to 5-7) Methyl paraben 0.2% Purified water q.s. to 100g

[0598] (viii) Topical Gel 2 wt.% 'Compound X' 5% Methylcellulose 2% Methyl paraben 0.2% Propyl paraben 0.02% Purified water q.s. to 100g

[0599] (ix) Topical Ointment wt.% 'Compound X' 5% Propylene glycol 1% Anhydrous ointment base 40%

[0600] Polysorbate 80 2%

[0601] Methyl paraben 0.2%

[0602] Purified water q.s. to 100g

[0603] (x) Topical Cream 1 wt.% 'Compound X' 5% White bees wax 10% Liquid paraffin 30% Benzyl alcohol 5% Purified water q.s. to 100g

[0604] (xi ) Topical Cream 2 wt.%

[0605] 'Compound X' 5%

[0606] Stearic acid 10%

[0607] Glyceryl monostearate 3%

[0608] Polyoxyethylene stearyl ether 3%

[0609] Sorbitol 5%

[0610] Isopropyl palmitate 2 %

[0611] Methyl Paraben 0.2%

[0612] Purified water q.s. to 100g

[0613] These formulations may be prepared by conventional procedures well known in the pharmaceutical art. It will be appreciated that the above pharmaceutical compositions may be varied according to well-known pharmaceutical techniques to accommodate differing amounts and types of active ingredient 'Compound X'. Aerosol formulation (vi) may be used in conjunction with a standard, metered dose aerosol dispenser. Additionally, the specific ingredients and proportions are for illustrative purposes. Ingredients may be exchanged for suitable equivalents and proportions may be varied, according to the desired properties of the dosage form of interest.

[0614] All publications, patents, and patent documents cited herein are incorporated by reference as though individually incorporated by reference. No limitations inconsistent with this disclosure are to be understood therefrom. The invention has been described with reference to various specific and preferred embodiments and techniques. However, many variations and modifications may be made while remaining within the spirit and scope of the invention.

[0615] While specific embodiments have been described above with reference to the disclosed embodiments and examples, such embodiments are only illustrative and do not limit the scope of the invention. Changes and modifications can be made in accordance with ordinary skill in the art without departing from the invention in its broader aspects as defined in the following claims.

Claims

CLAIMSWhat is claimed is:

1. A compound of Formula I:wherein,W is CR1or N;R1is halo, substituted -(C1-C6)alkyl, or unsubstituted or substituted -(C2-C8)alkynyl;X is NRAor O;RAis H or -(C1-C6)alkyl;R2and R3are each independently H, halo, CN, NO2, or -(C1-C6)alkyl;R4is H, -(C1-C6)alkyl, or -(C=O)(Ci-C6)alkyl;Y is O or S;Z1is substituted or unsubstituted 4-, 5- or 6-membered nitrogen heterocycle, substituted phenyl, substituted -NH(C2-C6)alkyl, or substituted -(C2-C8)alkynyl; and m is 0 or 1; or a pharmaceutically acceptable salt thereof.

2. The compound of claim 1, wherein Z1is substituted or unsubstituted pyrrolidine- 1-yl, pyrrolidine-3 -yl, piperidine- 1-yl, or piperidine-4-yl.

3. The compound of claim 1, wherein Z1is:

4. The compound of claim 1, wherein the substituent on the substituted 4-, 5- or 6- membered nitrogen heterocycle, substituted -NH(C2-C6)alkyl, or substituted -(C2-C8)alkynyl is a diazirine substituent.

5. The compound of claim 1, wherein Z1is:whereinR5is -O(C2-C8)alkynyl, N3, or nitrogen-substituted -OCH2-triazole; andR6is H, -(C2-C8)alkynyl, -O(C2-C8)alkynyl, or nitrogen- substituted -OCH2 -triazole.

6. The compound of claim 5, wherein one of R5or R6is nitrogen-substituted -OCH2-tri azole and the triazole moiety of R6is:wherein n is 1-5 and p is 1-5.

7. The compound of claim 1, wherein m is 0.

8. The compound of claim 1, wherein m is 1.

9. The compound of claim 1, wherein the compound has an ( / ^-configuration at the indoIone stereocenter.

10. The compound of claim 1, wherein X is NH and Y is O; or W is CR1; or W is CR1, X is NH, and Y is O.

11. The compound of claim 1, wherein the compound is represented by Formula II:or a pharmaceutically acceptable salt thereof.

12. The compound of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

13. The compound of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

14. The compound of claim 1, wherein the compound is:or a pharmaceutically acceptable salt thereof.

16. A pharmaceutical composition comprising a compound of claim 1 and a pharmaceutically acceptable excipient.

17. A method for treating cancer in a patient in need thereof, comprising administering a therapeutically effective amount of a pharmaceutical composition according to claim 11, wherein the cancer is thereby treated.

18. The method of claim 17, wherein the compound in the pharmaceutical composition is:or a pharmaceutically acceptable salt thereof.

19. The method of claim 17, wherein the cancer is breast cancer.

20. The method of claim 17, wherein the therapeutically effective amount of the pharmaceutical composition administered comprises about 2 mg / kg to about 50 mg / kg of the compound.

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