WO2020232381A1 - INHIBITING MUTANT ISOCITRATE DEHYDROGENASE 1 (mIDH-1) - Google Patents

Abstract

Patients diagnosed with a cancer harboring an IDH-1 mutation can be treated by the administration of a therapeutically effective amount of a pharmaceutical composition comprising Compound 1, a selective inhibitor of 2-HG production from mIDH-1 enzymes including the R132 mutations R132C, R132H, R132L, R132G, and R132S.

Description

INHIBITING MUTANT ISOCITRATE DEHYDROGENASE 1 (mIDH-1)

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Application No.

63/023,813, filed May 12, 2020, U.S. Application No. 16/526,593, filed July 30, 2019, U.S. Application No. 16/431,588, filed June 4, 2019, U.S. Application No. 16/414,505, filed May 16, 2019, International Application No. PCT/US19/32747, filed May 16, 2019; the contents of each of the applications listed above are hereby incorporated herein by reference in their entirety.

TECHNICAL FIELD

[0002] The present disclosure relates to the treatment of cancer. In particular, the present disclosure provides methods of treating patients diagnosed with a cancer harboring certain mutant IDH-1 cancer cells.

BACKGROUND

[0003] Dysregulation of metabolism is a common phenomenon in cancer cells. The

NADP(+)-dependent isocitrate dehydrogenases 1 and 2 (IDH-1 and IDH-2) functionally modulate cellular metabolism in lipid synthesis, cellular defense against oxidative stress, oxidative respiration, and oxygen-sensing signal transduction. The presence of mutations in IDH-1 imparts a neomorphic activity to the enzyme, resulting in the production of (R)-2- hydroxyglutarate (2-HG), the downstream effects of which cause epigenetic changes that consequently block the proper differentiation of progenitor cells and lead to cancer. IDH-1 mutations have been reported in hematological malignancies, as well as many solid tumors types. By far the most frequent IDH-1 mutations occur at amino acid position R132, and include R132H, R132C, R132S, R132G, and R132L mutations.

[0004] Therapeutic compounds that can be useful for inhibition of mutant IDH-1 and/or mutant IDH-2 cancer cells (mIDH-1 and mIDH-2) are being developed for the treatment of certain cancers. These therapies may also reduce elevated 2-HG levels in these cancer patients. Many different small molecule inhibitors of mutant isocitrate dehydrogenase (mIDH) proteins with neomorphic activity are disclosed in publications (e.g., WO2016/044789, WO2016/044787, WO2016/044782, WO2016/171755, and WO2016/171756), including testing of these compounds in IDH-1 R132H and IDH-1 R132C enzymatic assays, and cellular 2-HG assay using HCT116 mutant IDH-1 cells.

[0005] Among the other mIDH-1 inhibitors that have been developed are those depicted in Table 1, which includes multiple compounds that have been reported in clinical trials, and certain other compounds that have been described as being useful for the treatment of cancer.

Table 1

soc ra e e y rogenase- 1

isocitrate dehydrogenase- inhibitor

1 inhibitor (Shanghai Haihe (Sanofi) Pharmaceutical)

ABT-199 [0006] The FDA recently approved the administration of 500 mg once daily of a mutant IDH1 inhibitor for the treatment of acute myeloid leukemia (AML) with a susceptible IDH1 mutation as detected by an FDA-approved test in (a) adult patients with newly-diagnosed AML who are > 75 years old or who have comorbidities that preclude use of intensive induction chemotherapy and (b) adult patients with relapsed or refractory AML. However, a 2017 Multi- Discipline Review panel at the FDA noted that doubling the drug dose of this compound translates to only approximately a 40% increase in exposure, and that an increase in clearance at steady state may be related to autoinduction. A PBPK model reasonably captured the autoinduction effect of the 500 mg QD dose of this compound on CYP3 A4, and described the steady-state PK profiles of this compound in patients at clinically relevant exposure levels. The fold-change in relative bioavailability of this compound from single-dose to steady-state for this compound was 0.50. In addition, following administration of a single oral dose of 50 mg/kg to rats with an intact blood-brain barrier, this compound exhibited brain penetration of 4.1% (AUC0-8h [brain] / AUC0-8h [plasma]).

[0007] There remains an unmet medical need for a therapeutically effective method of administering a mIDH-1 inhibitor providing the following benefits:

i) achieving sufficient predicted free brain drug exposure (e.g., a desirable predicted Cbrain Ratio as disclosed herein) suitable for treating mIDHI solid tumors in the central nervous system, including forms of brain cancer such as glioma; and

ii) achieving and sustaining a desired steady state drug plasma concentration within a suitable range for a patient throughout a desired course of treatment (e.g., at least six months).

[0008] Thus, there remain particular challenges associated with treating m-IDHl forms of cancer, including the treatment of mIDHI forms of blood cancer (e.g., AML) throughout a course of treatment (e.g., 15 days to 6 months), treatment of mIDHI forms of cancer across the blood brain barrier (e.g., mIDHI forms of glioma), and treatment of other mIDHI solid tumors.

[0009] There also remains a need for identifying therapeutic compounds that selectively inhibit the production of 2-HG from mIDH-1 cancer cells harboring R132 mutations including R132S, R132G and R132L. In addition, there remains a need for therapeutic compounds that selectively inhibit production of 2-HG from cancer cells harboring a variety of R132 IDH-1 mutations with clinically relevant comparative potencies, while remaining inactive at wild type IDH-1 cells. Preferably, a targeted, selective small molecule inhibitor of 2-HG production from mIDH-1 cancer cells is also inactive in mIDH-2 cancer cells that produce 2-HG. In addition, there is a need for inhibitors of the production of 2-HG from mIDH-1 cancer cells having a R132 mutation selected from the group consisting of: R132L, R132G, and R132S mutation in IDH-1.

SUMMARY

[0010] The present disclosure provides methods for treating cancer. The present disclosure encompasses the recognition that Compound 1 is useful for treating cancers harboring (i) an IDH1 mutation and (ii) a concurrent mutation (e.g., a mutation as described herein).

Accordingly, the present disclosure provides methods of treating cancers harboring (i) at least one IDHl mutation and (ii) at least one concurrent mutation.

[0011] The present disclosure identifies a particular mIDH-1 inhibitor (i.e., Compound 1) and, furthermore, provides therapeutic methods for using this compound in the treatment of cancer (e.g., AML, glioma, and various solid tumors). In the selection of the mIDH-1 inhibitor Compound 1, the present disclosure provides an insight that certain prior assessments of multiple mIDH-1 inhibitory compounds may have focused on and/or unduly prioritized one or more features (e.g., in vitro potency as measured in biochemical assays for the production of 2-HG) that can lead away from appreciation of certain unexpected properties of Compound 1 that led to the discovery of methods of treatment as described herein.

[0012] The discovery of the methods of treatment provided herein is based in part on the selection of Compound 1 from among many compounds reported to inhibit the production of 2- HG from mIDH-1 cancer cells. Compound 1 was not initially reported as having the greatest biochemical potency compared to reports for certain other small molecule inhibitors of mIDH-1. Structurally distinct compounds (e.g., AG-120, AG-881, IDH305, IDH889, GSK321, and Bayl436032) and other certain quinolinone-based compounds were initially reported as having greater in vitro potency in biochemical assays measuring activity against certain mIDH-1 isoforms. Compound 1 not only inhibits 2-HG production from cells harboring various R132X forms of IDH-1 mutations; Compound 1 is characterized by a CNS multiparameter optimization (“CNS MPO”) value supporting further development as an oral therapy for mIDH-1 forms of cancer in the central nervous system.

[0013] Compound 1 selectively inhibits production of 2-HG in mIDH-1 cancer cells (i.e., cancer cells harboring IDH-1 R132X mutations) with desirable in vitro potencies when compared to wild type IDH-1 cells and mIDH-2 cancer cells. In addition to this selectivity for mIDH-1, Compound 1 is active against multiple IDH-1 R132X mutants, and therefore can be used to treat patients diagnosed with cancers possessing a variety of such mIDHI mutations. Of these R132X IDH-1 mutants, R132H and R132C are the more frequently detected mutations for human mIDH-1 cancers. With respect to inhibiting 2-HG production from mIDH-1 R132C and R132H cell lines, Compound 1 shows comparable activity that is within 5-fold, compared to more disparate differences in activity ranging from about 8-fold to 240-fold for comparative compounds (Example 6).

[0014] In addition to R132H and R132C IDH-1 mutants, Compound 1 inhibits the IDH-1 mutants R132L, R132G, and R132S. Notably, Compound 1 inhibits all five of these IDH-1 mutants (R132L, R132G, R132S, R132H, and R132C) with only a 7-fold range in potencies. Therefore, patients diagnosed with cancer possessing mutant IDH-1, e.g., having an IDH-1 R132X mutation selected from the group consisting of: R132L, R132G, and R132S (in addition to R132H and R132C IDH-1 mutations), can be treated with Compound 1 (see, e.g., Example 3).

[0015] The present disclosure also provides suitable dosing regimens of Compound 1 for treating cancers harboring (i) an IDH1 mutation and (ii) a concurrent mutation. A suitable dose should possess an appropriate therapeutic index (e.g., an observed in vivo efficacy against multiple forms of cancer harboring mIDH-1, but without unacceptable levels of toxic side effects). More specifically, a drug plasma concentration providing in vivo efficacy (“Cefif”) in patients with tumors producing 2-HG has been defined in the literature as one that provides > 90% inhibition of 2-HG production (Fan et al., 2014). Based upon mouse xenograft experiments described in Example 4 and plasma protein binding correlation in humans, Compound 1 was found to have a Ceff of about 1,652 ng/mL. Therefore, preferred methods of administering Compound 1 to treat a patient having a cancer harboring mIDH-1 can achieve a plasma concentration of at least about 1,652 ng/mL.

[0016] In addition, preferred methods of administering Compound 1 avoid unacceptably high concentrations of Compound 1 in the patient. The discovery of the maximum preferred concentration of Compound 1 was based in part on the results from a 28-day oral toxicity study in monkeys (Example 10), which found that the most significant adverse event was an increase in mean QTc interval duration, a type of cardiac event that may cause arrhythmia. The lowest Cmax plasma concentration of Compound 1 at which prolonged QTc interval duration was observed was about 7,840 ng/mL. Accordingly, Compound 1 is preferably dosed in a manner that achieves a drug plasma concentration of no greater than about 7,800 ng/mL (“Ctox”).

[0017] Preferred methods of administering a therapeutically effective amount of Compound 1 provide a concentration of Compound 1 in the patient blood plasma within a therapeutic range of about 1,652-7,840 ng/mL.

[0018] The discovery of provided methods of administering Compound 1 is based on the evaluation of multiple doses of Compound 1 (100 mg, 150 mg and 300 mg) at multiple dose intervals (once daily and twice daily), both as a single agent and in combination with another therapeutic agent. Notably, the administration of Compound 1 at a total daily dose of 300 mg each day (preferably, 150 mg BID) ultimately demonstrated therapeutic benefits in patient treatment across multiple forms of mIDHI cancer. As described in Example 11, a Phase 1/2 study of Compound 1 was initiated to evaluate Compound 1 alone or in combination with azacitidine (“AZA”) in mIDH-1 AML/my elodysplastic syndrome (MDS) patients. The blood plasma concentration and other effects of administering Compound 1 were evaluated at multiple different doses of Compound 1 at different dose intervals: 100 mg QD (i.e., 100 mg once daily), 150 mg QD (i.e., 150 mg once daily), 300 mg QD (i.e., 300 mg once daily), and 150 mg BID (i.e., 150 mg twice daily), with Compound 1 administered both as a single agent and/or in combination with AZA. As shown in Figure 14 and described in Example 11, as a single agent, median Cmin concentrations of Compound 1 at steady-state were below Ceff in patients given doses of 100 mg or 150 mg once daily. When given at a dose of 300 mg once a day, Compound 1 achieved a median Cmin above Ceff, with a majority of patients reaching Ceff. However, 150 mg given twice daily achieved the highest median Cmin and also showed less inter-quartile variability in minimum concentrations as compared to 150 mg once daily.

[0019] Compound 1 was also administered in combination with azacitidine (AZA). As shown in Figure 15, in this combination, dosing a total of 150 mg of Compound 1 once daily shows a median Cmin less than 1000 ng/mL, which is well below Ceff. In contrast, dosing a total of 150 mg of Compound 1 twice daily shows a median Cmin of over 3000 ng/mL, which is nearly twice Ceff.

[0020] Therefore, the present disclosure provides that a total daily dose of 300 mg of Compound 1, preferably administered in a divided daily dose of 150 mg BID, is preferred compared to other doses and dose intervals tested. Applicant selected 300 mg total daily dose, administered as a 150 mg dose form. A suitable dose can be Compound 1 administered in a 300 mg dose once daily. Preferably, a suitable dose can be Compound 1 administered in a 150 mg dose twice daily.

[0021] Compound 1 provides an unexpectedly durable steady state drug plasma

concentration throughout a desired course of treatment. After the initial 15 days of treatment with 150 mg twice daily of Compound 1, the median steady state blood concentration of

Compound 1 was maintained above about 2,000 ng/mL throughout a course of treatment (e.g., up to about 36 weeks, including 12-32 weeks, as well as other intervals therein, all measured from initial administration of Compound 1). The median Css was also well below the predicted threshold for QTc prolongation risk as discussed above. As shown in Figure 18 A, which shows data for Compound 1 dosed as a single agent at 150 mg twice daily, the median Css achieved on day 15 of cycle 1 (i.e.,“C1D15, or on the 15th day of treatment with Compound 1) was maintained for at least six four-week treatment cycles, and in fact extended to beyond nine four- week cycles. While concentrations are somewhat variable across patients, the median Css level is maintained and does not trend downward. Good retention of Css levels were also observed in combination with AZA as shown in Figure 18B.

[0022] The discovery of the unique and remarkable properties of Compound 1 as a selective inhibitor of mIDH-1 across multiple mIDH-1 cancers led to the development of a dosing regimen of Compound 1 that overcomes therapeutic obstacles encountered with prior mIDH-1 inhibitor compounds. Through the administration of Compound 1 at a total daily dose of 300 mg, the present disclosure provides sustained delivery of a mIDH-1 inhibitor at a desired drug plasma concentration for the treatment of cancer in patients harboring mIDH-1, optionally harboring mIDHI and a concurrent mutation.

[0023] In some embodiments, the present disclosure encompasses the recognition that Compound 1 is useful for treating central nervous system (CNS) cancers, such as glioma, harboring (i) an IDH1 mutation and (ii) a concurrent mutation. IDH-1 mutations in brain cancers, such as glioma, can result in abnormal hypermethylation of histones and DNA and suppression of normal cellular differentiation. IDH-1 R132H mutations represent more than 90% of the IDH mutations present in low grade glioma and secondary glioblastoma multiforme (GBM) patients. In addition, IDH-1 mutations R132C and R132S are also reported in glioma patients. However, in order to be able to treat glioma, a small molecule inhibitor of mIDH-1 must be able to cross the blood brain barrier (“BBB”) at a therapeutically effective concentration over time, presenting another challenge to selecting a compound suitable for the treatment of glioma.

[0024] The ability to cross the BBB is by no means an intrinsic property of mIDH-1 inhibitors. Referring to Table 11, many known mIDH-1 inhibitors have undesirably low CNS MPO values (e.g., Table 11, compounds GSK321 and Bayl436032). Therefore, even among these advanced drug candidates, there is no apparent reported correlation between mIDH-1 inhibitory activity and CNS MPO values.

[0025] In fact, selecting a compound that is both a potent inhibitor of mIDH-1 and possesses a desirably high MPO value is not straightforward. For example, of the compounds specifically exemplified in WO2016/044789 (“the‘789 publication”), which include Compound 1, several compounds are reported as having greater in vitro potency than Compound 1 in at least one assay reported in Table 6 of the‘789 publication (i.e., compounds 1-20, 1-22, 1-23, 1-25, 1-26, 1-27, and 1-29). See Table 11, which reproduces these data from the‘789 publication. However, none of these compounds has a CNS MPO score as great as Compound 1. In fact, one compound (1-26) does not even meet a desired minimum MPO threshold value of 3.8 desired as a predictor of BBB permeability. Conversely, of the six compounds with CNS MPO scores higher than Compound 1, five are less potent in vitro than Compound 1 in at least one biochemical assay of IDH inhibition (i.e., compounds 1-2, 1-3, 1-5, 1-6, and 1-11), with one compound being “equipotent” (compound 1-1).

[0026] The lack of correlation between mIDH-1 inhibition and CNS MPO scores in Table 11 highlights the unpredictability in selecting a single mIDH-1 inhibitor compound that inhibits multiple R132X forms of mIDH-1 with sufficiently similar potencies, and is also characterized by a sufficiently high MPO score (e.g., 3.8 or higher), both of which are desired in a therapy to treat a mIDHI glioma. It is against this backdrop that the identification of Compound 1 as a compound having a MPO score of 3.8 or higher is unexpected. Indeed, rodent modeling showed a stark contrast between Compound 1 and two other reported mIDH-1 inhibitors, AG- 120 and AG-881. As described in Example 9, Compound 1 partitions into the brain at a level 2-fold greater than that estimated to achieve a therapeutic benefit, whereas AG-120 and AG-881 partition into the brain at a level less than what is estimated to achieve a therapeutic benefit.

Thus, even when compared to another compound also having a CNS MPO score suggestive of good BBB permeability, such as AG-881, these data indicate that Compound 1 possesses unexpectedly superior properties by combining desired comparative and selective in vitro potency and predicted drug exposure in the brain.

[0027] For instance, as described in Example 9, preclinical studies show that Compound 1 can cross the BBB in rodent models at desirable levels. Oral administration of Compound 1 showed high systemic bioavailabilty in multiple preclinical species. Permeability was excellent, with little evidence of efflux, and significant brain penetration was observed in mice (98% brain binding in murine animal model). Based on these assessments in rodents, Compound 1 is believed to cross the blood-brain barrier to an extent effective to reach free concentration levels in the brain consistent with pharmacological activity.

[0028] The present disclosure provides, among other things, methods of treating cancers harboring a mutation in IDH-1 (and optionally a concurrent mutation described herein). In particular, patients diagnosed with cancer harboring a mutant IDH-1 cancer cell, e.g., having a IDH-1 R132 mutation selected from the group consisting of: R132L, R132G, and R132S (in addition to R132H and R132C IDH-1 mutations), can be treated with a therapeutically effective amount of Compound 1. In some examples, patients treated with Compound 1 can have a mutant IDH-1 cancer that does not have a mIDH-2 mutation detected with a FDA approved mIDH-2 diagnostic (e.g., as provided at www.fda.gov/CompanionDiagnostics).

[0029] The patient can be diagnosed with a cancer (e.g., a hematologic malignancy such as MDS or AML) characterized by the presence of a mutant allele of IDHl (e.g., a mIDHI selected from the group consisting of: R132L, R132G, and R132S) and a concurrent mutation selected from the group consisting of FLT3, NPM1, CEBPA and TP53. Preferably, the cancer is not characterized by an IDH2 mutation. The patient can be treated with a therapeutically effective amount of Compound 1 (preferably, 150 mg of Compound 1 administered twice per day, each day) throughout a course of treatment (preferably, at least 6 months) as a single agent or in combination with another agent for treating the cancer (e.g., azacitidine).

[0030] Compound l is a small molecule inhibitor of mutated forms of isocitrate

dehydrogenase 1 (IDH-1) enzyme. Compound 1 targets the mutant IDH-1 variants R132L, R132G, and R132S at lower concentrations than the wild-type IDH-1 enzyme or mutant IDH-2 enzymes tested in vitro as disclosed herein. Compound 1 is useful for the treatment of adult patients diagnosed with cancer having an IDH-1 mutation as detected by an FDA-approved test. Compound 1 can be administered to patients in need thereof in a therapeutically effective amount (e.g., 150 mg orally twice daily until disease progression or unacceptable toxicity). Patients for the treatment of cancer with Compound 1 can be selected based on the presence of IDH-1 mutations in the blood or bone marrow. In one embodiment, the recommended starting dose of Compound 1 is 150 mg taken orally twice daily with or without food until disease progression or unacceptable toxicity. For patients without disease progression or unacceptable toxicity, the patient can receive the therapeutically effective amount of Compound 1 for a minimum of 6 months to allow time for clinical response.

[0031] The disclosure is based in part on the discovery that Compound 1 selectively inhibits the production of 2-HG from mIDH-1 cancer cells harboring R132 mutations including R132S, R132G and R132L with clinically relevant comparative potencies, while remaining inactive at wild type IDH-1 cells. In addition, Applicant has discovered that Compound l is a targeted, selective small molecule inhibitor of 2-HG production from mIDH-1 cancer cells and is also inactive in mIDH-2 cancer cells that produce 2-HG (e.g., Compound 1 selectively inhibits the production of 2-HG from mIDH-1 cancer).

BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 illustrates Compound 1 binding with mIDH.

[0033] Figure 2A and Figure 2B are each a schematic of diaphorase-coupled assays used in Example 1, which measure activity by the determination of the level of remaining co-substrate NADPH after the enzymatic reaction is quenched.

[0034] Figure 3A is a graph showing the results from a surface plasmon resonance (SPR) biophysical characterization of the molecular interaction between mIDH-1 inhibitor Compound 1 and recombinant IDH-1-R132H protein.

[0035] Figure 3B is a comparator graph showing the SPR characterization of Compound 1 at a BCL6 control surface.

[0036] Figure 4 consists of 4 panels: (a), (b), (c), and (d). Panel (a) illustrates free concentration of Compound 1 in plasma after three-dose oral administration (12.5, 25 and 50 mg/kg) with 12 hr dosing interval in mouse HCT116-IDH1-R132H/+ xenograft model. Panel (b) illustrates free concentration of Compound 1 in tumor after three-dose oral administration (12.5, 25 and 50 mg/kg) with 12 hr dosing interval in mouse HCT116-IDH1-R132H/+ xenograft model. Panel (c) illustrates percent 2-HG inhibition in tumors in a PO dose of 12.5 mpk, 25 mpk and 50 mpk at three different time points (4h, 12h, 24h). Panel (d) illustrates in vivo activity (2- HG % inhibition) of Compound 1 vs free compound concentration in tumor.

[0037] Figure 5 is a synthetic reaction scheme for the preparation of Compound 1.

[0038] Figure 6 is a graph showing plasma and brain exposure for Compound 1 following 5 mg/kg PO dose in a CD-I Mouse.

[0039] Figure 7 is a graph showing plasma and brain exposure for Compound 1 following 100 mg/kg PO dose in a CD-I Mouse.

[0040] Figure 8 A illustrates the summary of cohorts from a phase 1 study in mIDHI AML.

[0041] Figure 8B illustrates use of Compound 1 in a phase 2 study in mIDHI AML and

MDS.

[0042] Figure 9A shows clinical response of patients in a human clinical trial in mIDHI AML who received Compound 1 monotherapy.

[0043] Figure 9B shows clinical response of patients in a human clinical trial in mIDHI AML who received Compound 1 and azacitidine combination therapy.

[0044] Figure 10 shows survival of patients in a human clinical trial in mIDHI AML.

[0045] Figure 11 illustrates frequency of baseline co-mutations in AML patients from

Example 11.

[0046] Figure 12 shows time on treatment for patients in a human clinical trial for mIDHI MDS.

[0047] Figure 13 illustrates frequency of baseline co-mutations in MDS patients from Example 11.

[0048] Figure 14 is a graph of the minimum blood plasma concentration (Cmin) of

Compound 1 measured in groups of human patients receiving Compound 1 as a single agent at different dose amounts and dose intervals:day 15 trough (ng/mL) of Compound 1 after administration to patients of 100 QD, 150 mg QD, 300 mg QD, and 150 mg BID Compound 1 as a single agent. Figure 14 also illustrates steady state is achieved by Week 2; (tl/2 ~ 60 hrs) and 150 mg BID and 300 mg QD steady state exposures are > Ceff

[0049] Figure 15 is a graph of the minimum blood plasma concentration (Cmin) of

Compound 1 measured in groups of human patients receiving Compound 1 at different dose amounts and dose intervals, in combination with azacitidine: day 15 trough (ng/mL) of

Compound 1 after administration to patients of 150 mg QD and 150 mg BID of Compound 1 in combination with azacitidine. Figure 15 also illustrates steady state is achieved by Week 2; (tl/2 ~ 60 hrs) and 150 mg BID QD steady state exposures are > Ceff.

[0050] Figure 16 illustrates a therapeutically reduced level of 2-HG in the patient plasma after consecutive treatment cycles of treatment with Compound 1 as a single agent.

[0051] Figure 17 illustrates a therapeutically reduced level of 2-HG in the patient plasma after consecutive treatment cycles of treatment with Compound 1 and azacitidine.

[0052] Figure 18A is a graph of blood plasma concentration of Compound 1 measured in a group of human patients treated with Compound 1 as a single agent throughout a course of treatment. The dashed line labeled“Ceff’ is lower than 1652 ng/mL.

[0053] Figure 18B is a graph of blood plasma concentration of Compound 1 measured in a group of human pateints treated with Compound 1 and azacitidine throughout a course of treatment. The dashed line labeled“Ceff’ is lower than 1652 ng/mL.

[0054] Figure 18C is a graph of effective blood plasma concentration of Compound 1 measured in a group of human patients throughout a course of treatment.

[0055] Figure 18D is a graph of blood plasma concentration of Compound 1 measured in a group of human patients treated with Compound 1 and azacitidine throughout a course of treatment. The dashed line labeled“Ceff’ is lower than 1652 ng/mL.

[0056] Figure 18E is a graph of blood plasma concentration of Compound 1 measured in a group of human patients throughout a course of treatment.

[0057] Figure 19A is a graph of the level of 2-HG in plasma of a group of human patients after consecutive treatment cycles of treatment with Compound 1 as a single agent.

[0058] Figure 19B is a graph of the level of 2-HG in plasma of a group of human patients after consecutive treatment cycles of treatment with Compound 1 and azacitidine.

[0059] Figure 19C is a graph of the level of 2-HG in plasma of a group of patients after two consecutive treatment cycles.

[0060] Figure 19D is a graph showing the reduced level of 2-HG measured over time in a group of human patients treated with 150 mg of Compound 1 BID; the PD response is sustained throughout treatment with 150 mg of Compound 1 BID.

[0061] Figure 20A is a graph showing the levels of steady-state concentration of Compound 1 measured in a group of human patients at the day 15 trough (ng/mL) of Compound 1 after administration to patients of 150 mg QD, 300 mg QD, and 150 mg BID of Compound 1. Figure 20A also illustrates steady state is achieved by Week 2; (tl/2 ~ 60 hrs) and 150 mg BID steady state exposures > IC90 and below levels are expected to increase QTcF potential.

[0062] Figure 20B is a graph showing the 2-HG level measured in plasma (ng/mL) in a group of human patients after administration of 150 mg QD, 300 mg QD, and 150 mg BID of Compound 1.

[0063] Figure 20C is a graph showing the plasma 2-HG reduction observed in a human patient population at all doses and schedules by cycle 2, day 1 of administration of Compound 1 and that a preferred PD response was observed with 150 mg BID of Compound 1.

[0064] Figure 21 A is a graph Css over time of patients treated with Compound 1 at 150 mg

BID. Figure 21 A also illustrates that plasma exposures of Compound 1 are stable throughout treatment duration in patients. This can serve as basis for selecting a 150 mg BID dose for dose expansion and a recommended phase II dose.

[0065] Figure 21B is a graph plotting the ratio of steady state blood concentration of Compound 1 measured at different points during a Course of Treatment in a human patient during the administration of Compound 1 (150 mg BID). The Y-axis values are normalized to 1.0 using the concentration measured on day 15 of a Course of Treatment. The data for this graph were obtained from a single patient who received 150 mg BID of the solid form of Compound 1 obtainable from Example 5 throughout a Course of Treatment of over 300 days (i.e., greater than 6 months).

[0066] Figure 22A is a graph showing the correlation between Compound 1 and 2-HG plasma levels in patients treated with Compound 1 as a single agent across treatment groups and irrespective of time on treatment.

[0067] Figure 22B is a graph showing the correlation between Compound 1 and 2-HG plasma levels in patients treated with Compound 1 and azacitidine in combination across treatment groups and irrespective of time on treatment.

[0068] Figure 23A, Figure 23B, and Figure 23C are graphs showing time on treatment of AML patients treated with a single agent (Compound 1).

[0069] Figure 24A and Figure 24B are graphs showing time on treatment of AML patients treated with a combination of Compound 1 and azacitidine.

[0070] Figure 25 is a graph showing strong correlation between ddPCR and NGS in AML patients from Example 11. [0071] Figure 26 is a graph showing good concordance in VAF between bone marrow analysis (BMA) and white blood cells (WB) in AML patients from Example 11.

[0072] Figure 27A and Figure 27B illustrate change in IDH1 VAF across categories.

[0073] Figure 28 is a graph showing that clinical response in a treatment naive (TN) AML patient treated with Compound 1 in combination with azacitidine is associated with decrease in 2-HG and clearance of the IDHlm clone.

[0074] Figure 29 is a graph showing that clinical response in a R/R AML patient treated with Compound 1 as a single agent is associated with decrease in 2-HG and clearance of the

IDHlm clone.

[0075] Figure 30 is a graph showing that clinical response in MDS patients treated with Compound 1 is associated with a decrease in 2-HG and mutation clearance.

[0076] Figure 31 is a graph showing IDH2 -mediated resistance.

[0077] Figure 32 is a graph showing that presence of additional non-IDHlm clones drive resistance.

[0078] Figure 33 shows duration of Compound 1 monotherapy treatment in predominantly enhancing gliomas. ^a Patient did not have measurable disease and was not included in efficacy analysis. *Patients stayed on study with PD due to clinical benefit.

[0079] Figure 34 shows best percent change in tumor burden with Compound 1

monotherapy per investigator assessment (RANO).

[0080] Figure 35 shows percent change in tumor burden with Compound 1 monotherapy per central volumetric assessment (BCIR).

[0081] Figure 36 illustrates frequency of baseline co-mutations in glioma patients from Example 12.

[0082] Figure 37 shows Compound 1 plasma concentrations in glioma patients (n=24) from a human clinical trial.

[0083] Figure 38A shows baseline and maximum reduction plasma levels of 2-HG (after a minimum of 28 days of Compound 1 treatment) for 15 disease-evaluable patients with paired samples.

[0084] Figure 38B shows maximum percent change from baseline of 2-HG (after a minimum of 28 days of Compound 1 treatment) for 15 disease-evaluable patients with paired samples. [0085] Figure 39 shows Compound 1 plasma concentrations in glioma patients (n=32) from a human clinical trial.

[0086] Figure 40 shows duration of Compound 1 monotherapy treatment in glioma patients from a human clinical trial. ^a Patient did not have measurable disease and was not included in response analysis. ^b Patient discontinued after one dose due to adverse event, was non-evaluable, and was not included in clinical activity analyses.

[0087] Figure 41 shows % change from baseline in investigator assessment per RANO. ^a Changes >100% are shown as 100%.

[0088] Figure 42 shows % change from baseline for central volumetric assessment (BICR). ^a Changes >100% are shown as 100%.

[0089] Figure 43 shows a timeline of a patient’s therapy, including exemplary brain scans throughout the patient’s treatment with Compound 1. Percentage change from baseline in tumor size as assessed by the investigator is given below each scan.

DEFINITIONS

[0090] As used herein, the term“Course of Treatment” refers to the time period in which a patient is being administered an agent, including any administration holidays or recovery periods. A course of treatment can include a single treatment cycle or multiple treatment cycles. Additionally, a course of treatment can include a partial treatment cycle. A Course of Treatment can include the total time period during which a patient is on a treatment protocol for a disease, e.g. AML or MDS, with a therapy comprising the administration of a mIDH-1 inhibitor compound.

[0091] “Next-generation sequencing or NGS or NG sequencing” as used herein, refers to any sequencing method that determines the nucleotide sequence of either individual nucleic acid molecules (e.g., in single molecule sequencing) or clonally expanded proxies for individual nucleic acid molecules in a high-throughput fashion (e.g., greater than 103 or more molecules are sequenced simultaneously). Various next generation sequencing methods are known. In one embodiment, the relative abundance of the nucleic acid species in the library can be estimated by counting the relative number of occurrences of their cognate sequences in the data generated by the sequencing experiment. Next generation sequencing methods are known in the art, and are described, e.g., in Metzker, M. (2010) Nature Biotechnology Reviews 11 :31-46, incorporated herein by reference. Next generation sequencing can detect a variant present in less than 5% of the nucleic acids in a sample. As shown in Example 11, for the purposes of Variant Allele Frequency analysis, digital droplet PCR (ddPCR) can also be used. ddPCR methods are known in the art, and are described in, e.g., Hindson B. J., et al. (2011). High-throughput droplet digital PCR system for absolute quantitation of DNA copy number. Anal. Chem. 83(22): 8604-8610, and Volegstein, B., et al. (1999) Digital PCR. Proc. Natl. Acad. Sci. USA 90: 9236-9241, incorporated herein by reference.

[0092] As used herein, the term“R132X mIDH-1 mutation(s)” refers to a mutation at the IDH-1 arginine 132 that results in inhibitory activity of Compound 1 against the mutated IDH-1 form harboring the R132 mutation. Preferably, the R132X mutations have a 2-HG IC50 value of less than 500 nM (most preferably less than 250 nM or less than 150 nM) using the in vitro assay of Example 1. Accordingly, preferred R132X mutations include R132H and R132C, as well as R132L, R132G, and R132S (or other R132X mutations having therapeutically relevant 2-HG IC50 values obtained using the in vitro assay of Example 1). Patients having R132X mIDH-1 mutation(s) can be identified using a suitable diagnostic, such as a diagnostic analyzing patient tissue with next generation sequencing technology that identified the presence of the R132X mIDH-1 mutation in the patient tissue sample.

[0093] As used herein, the term“R132X mIDH-1 Selective Inhibitor Therapy” refers to a therapy administered to a patient to inhibit the activity of R132X mIDH-1 in the patient, where the therapy is known to have selective inhibitory activity against R132X mIDH-1 over wild type IDH-1. An R132X mIDH-1 selective inhibitor therapy can be administration of Compound 1 as disclosed herein.

[0094] As used herein, "sequencing" can be Next Generation Sequencing (NGS), a high- throughput sequencing technology that performs thousands or millions of sequencing reactions in parallel. Although the different NGS platforms use varying assay chemistries, they preferably generate sequence data from a large number of sequencing reactions run simultaneously on a large number of templates. The sequence data can be collected using a scanner, and then assembled and analyzed bioinformatically. Thus, the sequencing reactions are performed, read, assembled, and analyzed in parallel.

[0095] The terms“subject” and“patient” are used interchangeably in the present disclosure. [0096] Susceptible IDH1 mutations are defined as those leading to increased levels of 2- hydroxyglutarate (2-HG) in the specified mIDHI cancer cells (e.g., mIDHI leukemia cells or mIDHI glioma cells) and where efficacy is predicted by 1) clinically meaningful remissions with the recommended dose of Compound 1 and/or 2) inhibition of mutant IDHl enzymatic activity at concentrations of Compound 1 sustainable at the recommended dosage according to validated methods. Susceptible mutations include R132H and R132C mIDHI substitution mutations. In some methods, a susceptible IDHl mutation leads to increased levels of 2-hydroxyglutarate (2- HG) in the leukemia cells. In some methods, efficacy of Compound 1 is predicted by a) clinically meaningful remissions with the recommended dose of Compound 1 and/or b) inhibition of mutant IDHl enzymatic activity at concentrations of Compound 1 sustainable at the recommended dosage according to validated methods.

DETAILED DESCRIPTION

[0097] Compound l is a small molecule mIDH-1 inhibitor useful for the treatment of patients harboring IDH-1 mutations, in both hematologic and solid tumors. Compound 1 is also useful for treating patients harboring an IDH-1 mutation and a concurrent mutation.

(Compound 1)

[0098] Compound 1 has potent and equivalent biochemical activity against a number of IDH-1 arginine 132 (R132) mutated forms, of which R132H and R132C are the most prevalent observed for human IDH-1. Compound 1 is a small molecule mIDH-1 (mutated isocitrate dehydrogenase 1) inhibitor. It is a permeable, orally bioavailable compound, with an excellent preclinical profile in both in vitro and in vivo models.

[0099] Isocitrate dehydrogenase (IDH) is a class of enzymes that catalyze the oxidative decarboxylation of isocitrate to a-keto-glutarate (a-KG). There are three isoforms in human cells. IDH-1 resides in the cytosol and peroxisomes, whereas IDH-2 and IDH-3 are mitochondrial enzymes. IDH-1 is dimeric and uses NADP+ as an electron acceptor. IDH-3 is a tetrameric enzyme and, in contrast, uses NAD+ as an electron acceptor. IDH-3 is the primary IDH enzyme participating in the Krebs cycle. The presence of the IDH-1 mutations imparts a neomorphic activity to the enzyme, resulting in the production of (R)-2-hydroxyglutarate (2-HG) which has been termed an“oncometabolite”, and has pleotropic roles in tumorgenesis.

[0100] Since IDH-1 mutations are only found in tumor tissue, the present disclosure is based in part on the discovery that the selective mIDH-1 inhibitor of Compound 1 can be developed as a targeted therapy for multiple mIDH-1 forms of cancer. A patient selection biomarker for the use of Compound 1 can be the existence of IDH-1 mutation in a patient diagnosed with a cancer harboring mIDH-1. Studies in genetically engineered mouse models and models derived from cancer patient samples both support the discovery that mIDH produces 2-HG, the downstream effects of which cause epigenetic changes that consequently block the proper differentiation of progenitor cells and lead to cancer. In particular, IDH-1 mutations can lead to the loss of wild type enzymatic activity (conversion of isocitrate to alpha-KG (a-KG)). Instead, the mutated enzymes acquire the neomorphic activity of converting a-KG to 2-HG. In mIDH-1 harboring cancer cells, wild type and mutant IDH-1 form a heterodimeric complex that can produce very high 2-HG levels. All IDH-1 mutations result in the formation of the (R)-enantiomer of 2-HG, which is in contrast to the accumulation of (S)-enantiomer found in L2-HG aciduria patients, who harbor homozygous loss-of-function mutations in 2-HG dehydrogenase. Given the structural similarity between 2-HG and a-KG, 2-HG has been shown to be a competitive inhibitor of a number of a-KG dependent histone and DNA demethylases. 2-HG inhibits several KDM family histone demethylases in vitro, including H3K9/H3K36 demethylases KDM4A and KDM4C, and H3K36 demethylase KDM2A. Furthermore, elevated methylation levels of H3K4, H3K9, H3K27, and H3K79 have been observed in mIDH-1 containing patient-derived samples, as well as in cells expressing IDH mutations or treated with a cell-permeable ester of 2-HG. 2- HG also inhibits the TET family of DNA demethylases, which in turn results in the

hypermethylation of DNA CpG islands. Mutations in IDH- 1/2 and TET2 are thus far mutually exclusive, which supports the notion that 2-HG produced by mIDH inhibits TET2 and impairs hematopoietic cell differentiation. In addition, 2-HG has also been shown to block PHD activity, which is critical for regulation of hypoxia inducible factors and collagen hydroxylation and maturation. Hydroxylated collagen is important for the regulation of proliferation and proper differentiation of hematopoietic cells in bone marrow. Mutated IDH is also reported to block proper hepatocyte differentiation and promote cholangiocarcinoma. Since IDH-1 mutations are only found in tumor tissue, the present invention is based in part on the discovery of that the selective mIDH-1 inhibitor of Compound 1 can be developed as a targeted therapy for cancer. The patient selection biomarker for the use of Compound 1 can be the existence of IDH-1 mutation in a patient diagnosed with a cancer harboring mIDH-1.

[0101] Using in vitro cellular mechanistic assays monitoring levels of the errantly

overproduced, tumorigenic metabolic byproduct 2-hydroxy glutarate (2-HG), inhibition of mIDH-1 results in a >90% reduction in levels of measured 2-HG, an effect that has also been shown to translate into similar levels of 2-HG suppression in in vivo PK-PD studies in HCT116 (IDH-1 R132H) and HCT116 (IDH-1 R132C) xenograft bearing mice. In both models, the free concentration of Compound 1 was comparable in plasma and xenograft tumors, and exposures were dose dependent. At the highest dose tested in these studies (50 mg/kg), Compound 1 inhibited 2-HG levels in tumor by >90% for up to 24 hours after the last dose in the HCT116 (IDH-1 R132H) xenograft model, and to similar levels for at least 12 hours in the HCT116 (IDH- 1 R132C) model.

[0102] Accordingly, Compound 1 is useful in methods of treating patients diagnosed with a cancer harboring an IDH-1 mutation. The neomorphic enzymatic activity acquired as a result of IDH-1 mutation is believed to lead to the conversion of a-ketoglutarate (alpha-KG) to 2- hydroxyglutarate (2-HG). In consequence, patients bearing IDH-1 mutations have elevated levels of 2-HG. Most IDH-1 mutations result in a single amino acid change at the R132 residue, whereas most IDH-2 mutations occur at either Arginine 140 (R140) or Arginine 172 (R172). The IDH mutation spectrum varies among different tumor types (Table 2).

Table 2

[0103] For example, IDH-1 R132 mutations represent more than 90% of the IDH mutations present in low grade glioma and secondary GBM patients. IDH-1 mutations have been reported in hematological malignancies such as acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS), as well as many solid tumor types, including low grade glioma, secondary glioblastoma, intrahepatic cholangiocarcinoma (IHCC), chondrosarcoma, and melanoma. By far the most frequent IDH-1 mutations occur at amino acid position R132, and include R132H, R132C, R132S, R132G, and R132L mutations. Given that Compound 1 is a potent inhibitor of a spectrum of different IDH-1 R132 mutations, but is inactive against either wild type IDH-1 or mutated IDH-2, patients will be selected based on the occurrence of an IDH-1 mutation at the R132 residue.

[0104] The patient can be diagnosed as having an IDH-1 R132 mutation disclosed herein using sequencing methods, such as next-generation sequencing methods. The diagnostic patient selection method can be a next-generation sequencing (NGS)-based tumor genotyping assay analyzing a patient tissue sample such as a bone marrow sample. Useful techniques and technologies for diagnosing a patient as having a IDH-1 R132 mutation may include, without limitation, sequencing machines and/or strategies well known in the art, such as those developed by Illumina/Solexa (the Genome Analyzer; Bennett et al. (2005) Pharmacogenomics, 6:373-20 382), by Applied Biosystems, Inc. (the SOLiD Sequencer; solid.appliedbiosystems.com), by Roche (e.g., the 454 GS FLX sequencer; Margulies et al. (2005) Nature, 437:376-380), and by others.

Co-mutations

[0105] In some embodiments, the present disclosure provides methods of treating a patient diagnosed with a cancer characterized by (i) an IDH1 mutation and (ii) a concurrent mutation, comprising orally administering 150 mg BID (i.e., 150 mg twice daily) of Compound 1 to the patient in need thereof. In human clinical trials (e.g., as described in Example 11 and Example 12), at least one baseline concurrent mutation was detected in the vast majority of patients. The present disclosure encompasses the recognition that, while the presence of certain co-mutations can be useful in selecting patients for a particular treatment regimen and/or informing cancer pathology, patients diagnosed with cancer harboring both an IDH1 mutation and a concurrent mutation can be treated with Compound 1 without regard for the identity of the concurrent mutation, i.e., no trend in clinical response was observed in a human clinical trial of R/R AML patients (see Example 11).

[0106] In some embodiments, a cancer is a hematological malignancy (e.g., acute myeloid leukemia or myelodysplastic syndrome), glioma, hepatobiliary carcinoma, chondrosarcoma, intrahepatic cholangiocarcinoma, or a non-CNS solid tumor. In some embodiments, a cancer is acute myeloid leukemia or myelodysplastic syndrome. In some embodiments, a cancer is glioma.

[0107] In some embodiments, Compound 1 is administered as a single agent. In some embodiments, Compound 1 is administered in combination with another therapeutic agent (e.g., azacitidine).

[0108] In some embodiments, Compound 1 is administered every day for at least six months.

[0109] In some embodiments, an IDH1 mutation is selected from R132C, R132H, R132G,

R132S, and R132L. In some embodiments, an IDHl mutation is selected from R132G, R132S, and R132L.

[0110] In some embodiments, a concurrent mutation is selected from CEBPA, DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXL1, FLT3, STAG2, IDH2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1,

PTPN11, ATM, TP53, EZH2, SETBP1, GATA2, CBP, CUX1, GFAP, TERT, MGMT, lpl9q, OLIG2, ATRX, PI3K3CA, CDKN2B, CDKN2A, PTEN, and NogoA. In some embodiments, a concurrent mutation is selected from FLT3, NPM1, CEBPA, and TP53. In some embodiments, a concurrent mutation is selected from DNMT3A, NPMl, SRSF2, NRAS, RUNX1, ASXLl,

FLT3, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NFl, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM, and TP53. In some embodiments, a concurrent mutation is selected from CEBPA, DNMT3A, NPMl, SRSF2, NRAS, RUNX1, ASXLl, FLT3, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NFl, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM, and TP53. In some embodiments, a concurrent mutation is selected from NPMl, SRSF2, RUNX1, ASXLl, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NFl, ASXL2, EED, WT1, CBL, CSF3R, ETNK1,

PTPN11, ATM and TP53.

[0111] In some embodiments, a concurrent mutation is selected from SRSF2, ASXLl, RUNX1, DNMT3A, CSF3R, NPMl, STAG2, EZH2, U2AF1, SETBP1, NRAS, FLT3, GATA2, TP53, CBP, and CUX1. In some embodiments, a concurrent mutation is selected from SRSF2, ASXL1, RUNX1, DNMT3A, CSF3R, NPM1, STAG2, U2AF1, NRAS, FLT3, and TP53.

[0112] In some embodiments, a concurrent mutation is selected from TP53, GFAP, TERT, MGMT, lpl9q, OLIG2, ATRX, PI3K3CA, CDKN2B, CDKN2A, PTEN, NogoA, and

DNMT3 A. In some embodiments, a concurrent mutation is selected from TP53 and DNMT3 A. In some embodiments, a concurrent mutation is selected from TP53, ATM, and NRAS. In some embodiments, a concurrent mutation is selected from DNMT3 A, TP53, ATM, and NRAS.

[0113] In some embodiments, a concurrent mutation is not FLT3. In some embodiments, a concurrent mutation is not FLT3, CEBPA, BCOR, KRAS, NRAS, BCORLl, or DNMT3A. In some embodiments, a concurrent mutation is not IDH2.

[0114] In some methods, a therapeutically effective amount of Compound 1 can be administered to a patient having a mIDHI mutation at R-132 and a concurrent mutation at FLT3. The FMS-like tyrosine kinase 3 (FLT3) gene encodes a membrane bound receptor tyrosine kinase that affects hematopoiesis leading to hematological disorders and malignancies. FLT3 is one of the frequently mutated genes in hematological malignancies, such as adult acute myeloid leukemias (AML). The presence of a FLT3 internal tandem duplication has been detected in patients with acute myeloid leukemia (AML) and patients diagnosed with intermediate and high risk myelodysplastic syndrome (MDS). The heightened frequency of constitutively activated mutant FLT3 in adult AML has made the FLT3 gene a highly attractive drug target in this tumor type. A method for treating a FLT3 mutated mIDHI proliferative disorder can comprise identifying a mIDHI R132 mutation in a patient and measuring expression of a mutated FLT3 or a constitutively active FLT3 mutant, and one or more genetic abnormalities in a sample obtained from a tumor sample obtained from the patient; and administering to the patient a therapeutically effective amount of Compound 1 or a pharmaceutically acceptable salt thereof (e.g., 150 mg Compound 1 BID) for 6 months or more. Useful techniques and technologies for diagnosing a patient as having a IDH-1 R132 mutation may include, without limitation, sequencing machines and/or strategies well known in the art, such as those developed by Novartis (e.g.

LeukoStrat® CDx FLT3 (www.accessdata.fda.gov/cdrh_docs/pdfl6/pl60040c.pdf)).

[0115] A method of treating a patient with acute myeloid leukemia (AML), can comprise: (a) analyzing a genetic sample isolated from the patient for the presence of cytogenetic

abnormalities and a mutation in at least one of FLT3, NPMl, CEBPA, IDHl, and TP53 genes; and (b) treating the patient by administering a therapeutically effective amount of Compound 1 to the patient (e.g., a total of 150 mg of Compound 1 BID each day) if the mutation is present in R132 mIDHI and at least one of FLT3, NPM1, CEBPA and TP53 genes.

[0116] Compound 1 can be administered as a single agent as the R132X mIDH-1 Selective Inhibitor Therapy, or in combination with other therapeutic agents that are not mIDH-1 inhibitors as a combination for the R132X mIDH-1 Selective Inhibitor Therapy. As used herein, the term “R132X mIDH-1 mutation(s)” refers to a mutation at the IDH-1 arginine 132 that results in inhibitory activity of Compound 1 against the mutated IDH-1 form harboring the R132 mutation.

[0117] In some methods, Compound 1 is administered to a patient diagnosed as having a R132 IDH1 mutation either as a single agent or in combination with azacitidine. In some examples, patients have been treated with or are already being treated with azacitidine. In some embodiments, a combination therapy of Compound 1 and azacitidine can be administered for the treatment of patients with a cancer harboring aIDH-1 mutation (e.g., mIDHI forms of AML).

For example, patients can be administered Compound 1 daily (BID) in continuous 28-day cycles, in combination with azacitidine (administered at the dose of 75 mg/m2 for 7 days IV/SC per every 28-day cycle).

Achieving Effective Blood Plasma Concentration of Compound 1

[0118] The present disclosure provides methods for the treatment of cancer (e.g., AML or MDS or glioma) comprising a step of administering to a subject a therapeutically effective amount of a pharmaceutically acceptable form of Compound 1. In some examples, the pharmaceutically acceptable form of Compound 1 is an oral dosage form (e.g., as provided in Example 5) administered to the patient as R132X mIDH-1 Selective Inhibitor Therapy consisting of the oral administration of an oral dosage form of Compound 1 administered either as a single agent inhibitor of mIDH-1, or in combination with azacitidine or cytarabine. When Compound 1 is administered in such combination therapy, the subject can be receiving or have previously received treatment with azacitidine or cytarabine.

[0119] Post-therapy, 2-HG plasma levels less than 180 ng/mL are associated with better overall and disease-free survival in patients with IDH-1 and IDH-2 mutated AML (JANIN, M. et al., Serum 2-Hydroxyglutarate Production in IDHl- and IDH2 -Mutated De Novo Acute Myeloid Leukemia: A Study by the Acute Leukemia French Association Group, Journal of Clinical Oncology, 32(4): 297-305 (2014)). Figure 16 shows that 150 mg once daily dosing achieved a median 2-HG plasma concentration above 600 ng/mL. Meanwhile, 300 mg once daily achieved a median 2-HG plasma concentration slightly above 200 ng/mL, and 150 mg twice daily achieved a median 2-HG plasma concentration of approximately 100 ng/mL. This demonstrates the ability of 150 mg twice daily dosing to lower 2-HG levels below a literature established level as compared to 150 mg and 300 mg once daily.

[0120] The disclosure is based in part on the discovery that oral administration of Compound 1 (e.g., in the pharmaceutically acceptable oral dosage form resulting from the preparation method of Example 5) to humans having elevated blood 2-HG levels (i.e., above about 180 ng/mL) can provide a steady state (trough) blood concentration above a therapeutically effective amount of Compound 1 (e.g., above the IC90 concentration for R132H and/or R132C mIDH-1, and/or concentrations of greater than about 2,000 ng/mL or concentrations of greater than about 1652 ng/mL) throughout a course of treatment of 6 months from the initial administration of Compound 1, while simultaneously reducing and then maintaining the levels of 2-HG to below 200 ng/mL within about 15 days of daily treatment with Compound 1. Alternatively, the levels of 2-HG are maintained below 180 ng/mL within about 15 days of daily treatment with

Compound 1. In these observations, Compound 1 was administered in an oral dosage form (obtainable from the method in Example 5) twice per day (150 mg BID) to AML or MDS adult patients harboring a R132X IDH-1 mutation. After the initial 15 days of treatment with 150 mg BID of this oral dosage form of Compound 1, the steady state (trough) blood concentration of Compound 1 (pre-dose) was maintained above about 2,000 ng/mL (and well below the predicted threshold for QTc risk) throughout a course of treatment (e.g., up to about 30 weeks, including 12-30 weeks, and 20 weeks as well as other intervals therein, all measured from initial administration of Compound 1).

[0121] Accordingly, the present disclosure provides methods of treating patients harboring isocitrate dehydrogenase 1 mutations (mIDH-1) (preferably including one or more R132X mIDH-1) diagnosed with AML or MDS. The method can comprise administering to the patient in need thereof a therapeutically effective amount of a R132X mIDH-1 Selective Inhibitor Therapy. The R132X mIDH-1 selective inhibitor can consist of Compound 1 as the only R132X mIDH-1 inhibitor compound administered to the patient (e.g., in an oral dosage form such as the solid form obtained from Example 5). Compound 1 can be administered to a patient harboring the R132X mIDH-1 identified in a tissue sample, and/or an elevated 2-HG blood concentration (e.g., above about 180 ng/mL) for a course of treatment of at least three consecutive treatment cycles of 28 consecutive days of administration for each cycle. The course of treatment can start with the initial administration of Compound 1 in the first of the at least three or more consecutive 28-day treatment cycles. The administration of the therapeutically effective amount of

Compound 1 throughout a course of treatment (e.g., at least 15 consecutive days, preferably up to 30 weeks or more) to a patient having elevated 2-HG levels (e.g., 2-HG blood concentrations in plasma of 200-10,000 ng/mL) can result in a therapeutic effect on the patient evidenced by a durable therapeutically effective trough blood plasma concentration of Compound 1 in the patient throughout the course of treatment (e.g, above the IC90 concentration for R132H and/or R132C mIDH-1, and/or concentrations of greater than about 2,000 ng/mL and less than about 7,200 ng/mL, or above the IC90 concentration for R132H and/or R132C mIDH-1, and/or concentrations of greater than about 1652 ng/mL and less than about 7,840 ng/mL).

[0122] Compound 1 can be administered at a dose of 150 mg twice per day throughout the course of treatment. Compound 1 can be administered with food to improve bioavailability of Compound 1. The course of treatment can be at least 15 consecutive days starting with the initial dose of Compound 1 and longer (e.g., up to 30 weeks, 15 days to 30 weeks, 15 days to 12 weeks, at least 12 weeks, 12-30 weeks, 15 days to 6 months and other intermediate or longer durations or intervals apparent based on the present disclosure).

[0123] Some methods further comprise the administration of azacitidine to the patient throughout the course of treatment. Azacitidine can be subcutaneously or intravenously administered to the patient in an azacitidine treatment cycle consisting of the administration of a total dose of 75 mg/m2 each day for 7 consecutive days beginning at the start of each treatment cycle, followed by 21 consecutive days without administration of the azacitidine to the patient.

A 48-hour dose-interruption of azacitidine is allowed for weekends or holidays. If no response is seen after 2 treatment cycles, azacitidine can be administered at a total dose of 100 mg/m2 each day. Treatment with IDHlm inhibitor and azacitidine showed synergistic effects on releasing differentiation block in mIDH leukemia models in vitro.

[0124] The methods can further comprise the administration of cytarabine to the patient throughout the course of treatment. Cytarabine can be subcutaneously or intravenously administered to the patient in a cytarabine treatment cycle consisting of administration of a total dose of 20 mg/day each day for 7 consecutive days beginning at the start of each treatment cycle, followed by 10 consecutive days without administration of cytarabine to the patient. Cytarabine can also be administered 20 mg BID subcutaneously for 10 days beginning at the start of each treatment cycle. In the induction therapy of AML, the cytarabine dose administered in combination with other anticancer drugs can be 100 mg/m2/day by continuous IV infusion (Days 1 to 7) or 100 mg/m2 IV every 12 hours (Days 1 to 7). Cytarabine injection can be used intrathecally in acute leukemia in doses ranging from 5 mg/m2 to 75 mg/m2 of body surface area. The frequency of administration can vary from once a day for 4 days to once every 4 days. The dose can be 30 mg/m2 every 4 days until cerebrospinal fluid findings are normal, followed by one additional treatment.

[0125] A patient can be identified as having a R132X mutation in mIDH-1 using a diagnostic method comprising a sequencing analysis (e.g., next generation sequencing (NGS)) of bone marrow or other tissue sample obtained from the patient prior to the administration of Compound 1 to the patient. The R132X gene mutation can be determined prior to administration of

Compound 1 to the patient. Compound 1 can be administered to patients who have received prior anticancer therapy and/or other concomitant (non-anticancer) medications. In some examples, Compound 1 is administered to patient who has not received a prior mIDH-1 inhibitor therapy.

[0126] As provided herein, methods for the administration of an inhibitor of the R132X mutant IDH-1 (mIDH-1 Inhibitor) Compound 1 provide an unexpectedly durable steady state blood concentration of the mIDH-1 Inhibitor throughout a desired course of treatment. For example, therapeutic methods provided herein can provide AML or MDS patients harboring a R132X mIDH-1 mutation with durable steady state blood concentrations of the mIDH-1

Inhibitor of Compound 1 at a therapeutically effective level (e.g., above the IC90 concentration for a R312X mIDH-1) without a substantial decline (e.g., no more than 10% reduction) in initial Compound 1 mIDH-1 Inhibitor steady state blood concentration (e.g., blood concentration measured about 12 hours after an initial dose of the mIDH-1 Inhibitor) over a course of treatment of greater than about 12 consecutive weeks (e.g., 3 consecutive 28-day treatment cycles and preferably at least about 6 months). In addition, the mIDH-1 Inhibitor Compound 1 can be administered to AML or MDS patients harboring a R132X mIDH-1 mutation in a therapeutically effective manner that provides for the reduction of elevated 2-HG levels within about 15 days of initiating a course of treatment, preferably achieving and maintaining 2-HG levels in these patients at a level at or below about 180 ng/mL starting by day 15 in a course of treatment and continuing throughout a course of treatment lasting for at least 12 weeks or longer (e.g, 12-30 weeks).

[0127] Referring to Figures 18A-18E, the administration of Compound 1 in the oral dosage form described in Example 5 at 150 mg BID resulted in a sustained therapeutically effective trough blood plasma concentration above 2,000 ng/mL after cycle 3 of a 28-day treatment cycle. Simultaneously, referring to Figures 19A-19D, the administration of Compound 1 in the oral dosage form from Example 5 at 150 mg BID resulted in a sustained 2-HG level (e.g., under 200 ng/mL in plasma after cycle 3 day 1 of a 28-day treatment cycle). This effect is in contrast to other IDH1 inhibitors that have been advanced clinically. Accordingly, the invention is also based in part on the discovery that administration of Compound 1 at 150 mg BID results in a sustained ratio of greater than about 10 (preferably greater than about 20) of blood plasma concentration of Compound 1 (e.g., trough concentrations measured pre-dose at concentrations of about 2,000 ng/mL or greater) to 2-HG blood level (e.g, plasma concentrations of about 200 ng/mL or lower, including concentrations of about 100 ng/mL) after cycle 3 day 1 (i.e., after BID doses administered over the initial 15 consecutive days of treatment) of a 28-day treatment cycle. A plasma half-life of about 60 hours was estimated for Compound 1, with steady state achieved by week 2 of the course of treatment. The steady state blood concentrations of Compound 1 measured in the patients was above the IC90 value for 2-HG inhibition in R132X mIDH-1 cells (described in the Examples). As shown in Figures 18A-18E, the plasma exposures (steady state blood plasma concentration) of Compound 1 were durable (i.e., sustained) throughout the 30- week treatment duration. As shown in Figures 19A-19D the plasma 2-HG concentrations were reduced to the normal range within 1 cycle (C2D1) and maintained throughout the treatment duration. No dose limiting toxicities of Compound 1 were observed during dose escalation studies, and the maximum tolerated dose (MTD) of Compound 1 was not reached. Figures 18A- 18E are graphs of the data obtained from measuring the steady state concentrations from patients in the clinical trial receiving a 150 mg BID dose of Compound 1, either as a single agent or in combination with azacitidine as described in Example 11. As shown in Figure 18D, azacitidine did not alter the pharmacokinetics of Compound 1, with consistent Compound 1 plasma concentrations observed over the treatment duration. Figure 21 A is a graph showing the steady state concentration of Compound 1 measured in patients at various points during the Course of Treatment described in the clinical trial of Example 11, with each point representing a cycle number and day number (each cycle is 28 consecutive days of administration of 150 mg BID Compound 1). The steady state concentration of Compound 1 remained above the minimum desired level (bottom dashed line) and the maximum desired level (upper dashed line). Figure 2 IB is a graph plotting the ratio of steady state blood concentration of Compound 1 measured at different points during a Course of Treatment in a human patient during the administration of Compound 1 (150 mg BID). The Y-axis values are normalized to 1.0 using the concentration measured on day 15 of a Course of Treatment. The data for this graph were obtained from a single patient who received 150 mg BID of the solid form of Compound 1 obtainable from Example 5 throughout a Course of Treatment of over 300 days (i.e., greater than 6 months).

[0128] Compound 1, with a plasma half-life of ~60 hours, achieved steady-state

concentration within 2 weeks of dosing and remained consistent over treatment duration. At 150 mg BID (RP2D) as a single agent, steady-state plasma concentrations are above the preclinical Ceff resulting in > 90% reduction in plasma 2-HG and below Compound 1 levels predicted, in NHP, to pose a QTc prolongation risk. At 150 mg BID as a single agent, a significant reduction (p < 0.0009) in plasma 2-HG levels was achieved by end of Cycle 1 and was sustained over the treatment duration. Combination therapy of azacitidine plus Compound 1 150 mg BID achieved steady state concentrations of Compound 1 above the preclinical Ceff, resulting in > 90% reduction in plasma 2-HG and below Compound 1 levels predicted to pose a QTc prolongation risk. A significant (p < 0.0001) reduction in plasma 2-HG levels at the end of Cycle 1 was observed with the combination of Compound 1 (150 mg BID) and azacitidine and was sustained over the treatment duration. However, a slower rate of decline compared to the single agent Compound 1 has been observed.

[0129] The PK/PD relationship of individual subjects’ plasma Compound 1 and 2-HG concentration across single agent treatment groups and irrespective of time on treatment is presented in Figure 22A. Compound 1 concentrations < 1,000 ng/mL correspond to early time point assessments (C1D0 - C1D15). No correlation of exposure to 2-HG reduction is observed until Compound 1 concentrations crossed the Ceff predicted by in vivo models to result in > 90% reduction in plasma 2-HG levels. As shown in Figure 22B, no correlation of exposure to 2-HG reduction is observed until Compound 1 concentrations crossed the Ceff predicted by in vivo models to result in > 90% reduction in plasma 2-HG levels. Maximum, most consistent exposure-response was observed at the median Css of Compound 1 with 150 mg BID (RP2D).

[0130] The PK/PD relationship of individual subjects’ plasma Compound 1 and 2-HG concentration across combination of Compound 1 and azacitidine treatment groups and irrespective of time on treatment is presented in Figure 22B. Compound 1 concentrations <

I,000 ng/mL correspond to early time point assessments (C1D0 - C1D15). No correlation of exposure to 2-HG reduction is observed until Compound 1 concentrations crossed the Ceff predicted by in vivo models to result in > 90% reduction in plasma 2-HG levels. Maximum, most consistent exposure-response was observed at the median Css of Compound 1 with 150 mg BID (RP2D).

[0131] Figures 23A and 23C are graphs showing the results from the clinical trial in Example

I I, showing the 150 mg BID administration Compound 1 (solid form obtainable from Example 5) was administered as a single agent to multiple patients over a Course of Treatment with times having a median of 94 days, and a range of 1 to 586 days. About 32% of the patients remain on treatment.

[0132] Figure 23B depicts responses for relapsed or refractory (R/R) AML patients, and shows prolonged duration of treatment with Compound 1 was observed with a first response occurring within 2 months of treatment with Compound 1. Responses < CR/CRi were noted to deepen with continued treatment resulting in CR/CRh/CRi rate of 41% in R/R AML. Clinical benefit (SD > 8 weeks) was observed in subjects without an IWG defined response. 10% of patients (1 AML and 2 MDS) remained on treatment. Treatment Discontinuation: Progressive Disease (PD) (9), death (6), transplant (4), Adverse Events (AE) (3), investigator’s decision (2), withdrawal of consent (1), and lack of response (3).

[0133] Administration of Compound 1 at 150 mg BID in the clinical trial of Example 11 reduced the measured levels of 2-HG in the blood of patients as shown in Figures 19A-19D and Figures 20B-20C. The 2-HG levels measured in the blood of patients during this clinical trial of Example 11 are shown in Figures 19A-19D, demonstrating reduction of 2-HG levels within about 1-2 28-day treatment cycles (150 mg Compound 1 is administered BID on each day for 28 consecutive days for each treatment cycle, as described in Example 11).

[0134] Figures 20B-20C are each a graph showing the concentration of 2-HG measured in the blood of patients receiving one of three different doses and dose intervals: 150 mg QD, 300 mg QD, or 150 mg BID (either receiving Compound 1 as a single agent or in combination with azacitidine as described in the clinical trial of Example 11, in each category). The 2-HG levels are measured prior to administration of Compound 1, and then measured after administration of Compound 1 up to cycle 2, day 1 after first receiving Compound 1 (as the solid form obtained from Example 5).

[0135] This disclosure is based in part on the discovery that administration of Compound 1 at 150 mg BID resulted in a higher blood exposure level than either 150 mg QD or 300 mg BID at day 15. See, for example, Figure 20A. Administration of Compound 1 at 150 mg QD BID resulted in a blood exposure level of <3000 ng/mL at day 15. Administration of Compound 1 at 300 mg QD BID did not result in improved blood levels at day 15. In contrast, administration of Compound 1 at 150 mg BID results in a blood exposure level of >3000 ng/mL at day 15.

Additionally, the invention is based in part on the discovery that administration of Compound 1 at 150 mg BID resulted in a lower 2-HG level in plasma than either 150 mg QD or 300 mg BID at day 15. See, for example, Figure 20B.

[0136] The oral dosage form of Compound 1 (Example 5) was administered to human patients as a single agent (150 mg QD, 300 mg QD, 150 mg BID and 100 mg QD until disease progression) in a human clinical trial treating AML/MDS in cancer patients harboring a mIDHI mutation, as described in the Examples below. Figure 20A is a graph showing the concentration of Compound 1 measured in the blood of patients after receiving Compound 1 (as the solid form obtained from Example 5) in one of three different dose and dose intervals: 150 mg QD, 300 mg QD or 150 mg BID (either receiving Compound 1 as a single agent or in combination with azacitidine as described in the clinical trial of Example 11, in each category).

[0137] The present disclosure includes methods for treating AML or MDS in patients having one or more R132X mIDH-1 mutations (e.g., as measured in a tissue sample obtained from the patient) and/or elevated 2-HG levels (e.g., 2-HG levels measured in a blood sample at above about 180 ng/mL), comprising administration of Compound 1 alone (e.g., as a single agent) or in combination with azacitidine or cytarabine. For methods where Compound 1 is administered as a combination, the subject being administered Compound 1 may be receiving or previously received treatment with azacitidine or cytarabine.

[0138] The methods of treatment can include the administration of Compound 1 such that on day 1 of cycle 4 of repeated 28-day treatment cycles (or day 1 of any subsequent cycle), the trough blood plasma concentration of Compound 1 has not decreased more than about 5-25%, about 5-20%, about 5-15%, about 5-10%, about 10-25%, about 10-20%, or about 10-15%, as compared to the trough blood plasma concentration on day 1 of cycle 2. Preferably, patients harboring a R132X mIDH-1 mutation can be administered 150 mg of Compound 1 twice daily (BID) every day on consecutive days (without holiday) for one or more continuous 28-day cycles.

[0139] Compound 1 can be administered to certain patients in combination with a hypomethylating agent such as azacitidine. IDHl mutations (e.g., in AML or MDS patients harboring a R132X mIDH-1 mutation) can result in abnormal hypermethylation of histones and DNA and suppression of normal cellular differentiation. The combination of Compound 1 and azacitidine can be administered for the treatment of patients with AML harboring IDHl mutations. For example, patients can be administered the Compound 1 daily (BID) in continuous 28-day cycles, alone or in combination with azacitidine (administered at the dose of 75 mg/m2 for 7 days IV/SC per every 28-day cycle). For example, Compound 1 can be administered at a dose of 150 mg QD or 150 mg BID in combination with azacitidine

(azacitidine administered per standard of care for a patient). Figure 24A is a graph showing the results from the clinical trial in Example 11, showing the 150 mg BID administration Compound 1 (solid form obtainable from Example 5) in combination with the administration of azacitidine (administered at the dose of 75 mg/m2 for 7 days IV/SC per every 28-day cycle), administered to multiple patients over a Course of Treatment with times having a median of 87 days, and a range of 10 to 351 days. About 48% of the patients remain on treatment.

[0140] As shown in Figure 24B, upon further duration of treatment, clinical responses observed in R/R or TN AML and MDS: cytopenias associated with azacitidine may influence depth of IWG response. An ORR of 46% was observed for the combination of Compound 1 with azacitidine in R/R AML and an ORR of 78% was observed in TN AML (CR/CRi of 66%). Treatment Discontinuation was caused by: PD (6), transplant (6), investigator’s decision (5), death (4), AE (2), and others: treatment failure, hospice, other treatment (1 each). 37% of patients (AML and MDS) remain on treatment.

[0141] In some methods, Compound 1 can be administered with cytarabine. Low dose cytarabine (LDAC) can be administered to certain AML patients (e.g., AML patients at or above about 60 years of age who are not candidates for intensive therapy, and harboring a R132X mIDH-1 mutation). The therapeutically effective combination of Compound 1 with LDAC can be administered to AML patients harboring IDHl mutation. For example, patients can be administered the Compound 1 daily (BID) in continuous 28-day cycles, alone or in combination with LDAC (administered at the dose of 20 mg BID SC for 10 days every 28-day cycle) until treatment discontinuation.

[0142] Subjects that are treated according to provided methods and combination therapies can have relapsed or refractory AML or MDS, or“high risk” MDS, and may have been previously treated with a mIDHI inhibitor. Such refractory AML or MDS (i) can be naive to prior hypomethylating therapy and IDHl inhibitor therapy and/or (ii) may have shown inadequate response or progressed immediately preceding hypomethylating therapy. The provided methods and combination therapies can be used to treat subjects with residual IDH- R132 mutations. The provided methods and combination therapies can also be used to treat subjects with AML or MDS in morphologic complete remission or complete remission with incomplete blood count recovery (CR/CRi) after cytotoxic-containing induction therapy.

[0143] The methods of treatment are based in part on a human clinical study of

administration of Compound 1 in 3 stages: a phase 1 dose-escalation stage, a phase 1 dose- expansion stage and a phase 2 stage, as further described in the Examples. Single agent

Compound 1 dose escalation was administered in once-daily (QD) doses of 150 and 300 mg, a twice-daily (BID) dose of 150 mg or a once daily dose of 100 and 150 mg with food to potentially improve bioavailability. During the course of single agent dose escalation, a parallel escalation arm can be initiated for Compound 1 in combination with azacitidine. This combination can be initiated once the first dose level cohort of Compound 1 in the single agent schedule (150 mg QD) is complete. Once the maximum tolerated dose or the maximum evaluated dose is identified for the single-agent and combination cohorts, select populations of patients can be enrolled into phase 1 dose expansion cohorts at the selected single agent or combination doses, to further characterize the safety profile and confirm the recommended phase 2 dose. After the recommended phase 2 dose in combination with azacitidine is selected, a cohort of 6 patients are treated with Compound 1, at that dose, in combination with low dose cytarabine. In the Phase 2 portion, specific populations of patients with AML/MDS harboring IDHl -R132 mutations are enrolled to receive Compound 1 either as a single agent or in combination with azacitidine at the recommended phase 2 doses. [0144] As outlined in Examples 11 and 13, Compound 1 demonstrates clinical activity as single agent in a high-risk Phase 1 population of AML/MDS patients with IDHl mutation. 41% CR/CRh/CRi in R/R AML (35% in all AML/MDS) was observed in patients treated with Compound 1 as a single agent. Transfusion independence was observed in both IWG

responders/non-responders. Durable disease control or stable disease 4-12+ months was observed in R/R AML. An observed reduction in bone marrow blasts is supportive of clinical benefit to patients. Compound 1 was well tolerated with patients maintained in treatment for a median of 5.6 months, likely contributing to rate and depth (CR/CRh) of response. Compound 1 plasma exposure correlates with 2-HG response. Compound 1 Css reduction of 2-HG supports 150 mg BID as the dose and schedule selected for evaluation in global Phase 2 trial outlined in Figure 8B.

[0145] As outlined in Examples 11 and 13, the combination of Compound 1 and azacitidine demonstrates clinical activity in a high-risk Phase 1 population of AML/MDS patients with IDHl mutation. Patients maintained treatment for a median of 5 months. Durable disease control (> 6 months) was observed in the absence of IWG response. 46% ORR and 35% CR/CRh/CRi in R/R AML, 78% ORR in TN AML was observed. Compound 1 is well tolerated in

combination with azacitidine and possesses low risk of QT prolongation (2 AEs reported).

Azacitidine combination modestly increased metabolic and gastrointestinal treatment emergent adverse events (TEAEs). Higher rates of neutropenia compared to SA treatment (Grade 3/4 17% vs 6%) were observed which may be impacting the depth (CR/CRh) response. Compound 1 plasma exposure was shown to correlate with 2-HG response. Compound 1 Css reduction of 2- HG supports selection of 150 mg BID as RP2D.

[0146] In some embodiments, the present disclosure additionally provides methods of treating AML or MDS in a patient harboring isocitrate dehydrogenase 1 mutations (mIDHI), which can comprise administering to a patient in need thereof a therapeutically effective amount of Compound 1 each day for at least three consecutive treatment cycles of 28 consecutive days each. The administration of a therapeutically effective amount of Compound 1 can result in the patient having a durable therapeutically effective trough blood plasma concentration of

Compound 1 in the patient throughout the course of treatment.

[0147] In some embodiments, the administration of a therapeutically effective amount of Compound 1 can result in the level of 2-HG in the patient’s plasma being maintained at or below about 200 ng/mL at the start of the third consecutive treatment cycle (e.g., prior to dosing on day 15 or Cycle 3, Day 1), and the steady state blood plasma concentration of Compound 1 in the patient being maintained at or above about 2,000 ng/mL and below about 7,500 ng/mL

(preferably, below about 7,200 ng/mL) throughout the course of treatment. In some

embodiments, the steady state blood plasma concentration of Compound 1 in the patient is maintained at or above about 1652 ng /mL and below about 7,840 ng/mL throughout the course of treatment.

Maintaining Therapeutically Effective Blood Plasma Concentration of Compound 1 Throughout a Course of Treatment

[0148] The disclosure is based in part on the discovery that oral administration of Compound 1 (in the pharmaceutically acceptable oral dosage form resulting from the preparation method of Example 5) to humans can provide steady state (trough) blood concentrations above a therapeutically effective amount (e.g., above the IC90 concentration for R132H and/or R132C mIDH-1, and/or concentrations of greater than about 2,000 ng/mL) throughout a course of treatment of at least up to 30 weeks and beyond starting from the initial administration of Compound 1. Compound 1 can be administered to patients having elevated blood 2-HG levels (i.e., above about 180 ng/mL), leading to a reduction in 2-HG levels in the blood within 15 consecutive days starting with the first day of the administration of Compound 1, followed by maintaining the levels of 2-HG to below 200 ng/mL throughout the ensuing course of treatment. Alternatively, Compound 1 can be administered to patients having elevated blood 2-HG levels (i.e., above about 180 ng/mL), leading to a reduction in 2-HG levels in the blood within 15 consecutive days starting with the first day of the administration of Compound 1, followed by maintaining the levels of 2-HG to below 180 ng/mL throughout the ensuing course of treatment. Compound 1 was administered in an oral dosage form (obtainable from the method in Example 5) twice per day (150 mg BID). After the initial 15 days of treatment with 150 mg BID of this oral dosage form of Compound 1, the steady state (trough) blood concentration of Compound 1 (pre-dose) was maintained above about 2,000 ng/mL and well below the predicted threshold for QTc risk (e.g., below about 7,200 ng/mL) throughout a course of treatment (e.g., at least up to about 30 weeks or longer, including 12-30 weeks or up to 6 months or longer, from initial administration of Compound 1). Alternatively, after the initial 15 days of treatment with 150 mg BID of this oral dosage form of Compound 1, the steady state (trough) blood concentration of Compound 1 (pre-dose) was maintained above about 1652 ng/mL and well below the predicted threshold for QTc risk (e.g., below about 7840 ng/mL) throughout a course of treatment (e.g., at least up to about 30 weeks or longer, including 12-30 weeks or up to 6 months or longer, from initial administration of Compound 1). In some embodiments of the methods of treatment disclosed herein, the steady state blood concentration of Compound 1 after day 15 is maintained at greater than about 10-times the measured blood concentrations of 2-HG in the patient (e.g., at or below about 200 ng/mL). A R132X mIDH-1 Selective Inhibitor Therapy provides administering to a patient in need thereof a total dose of 150 mg BID of a pharmaceutically acceptable form of Compound 1 provided in Example 5 in an oral dosage form, on consecutive days throughout a Course of Treatment. Compound 1 is preferably the only inhibitor of mutant IDH-1 (mIDH-1) having one or more R132X mIDH-1 mutation(s) administered to the patient throughout the Course of Treatment of the R132X mIDH-1 Selective Inhibitor Therapy. Unless otherwise indicated, the mIDH-1 selective inhibitor (e.g. Compound 1) can be administered as a single agent as the R132X mIDH-1 Selective Inhibitor Therapy, or in combination with other therapeutic agents that are not mIDH-1 inhibitors as a combination for the R132X mIDH-1 Selective Inhibitor Therapy.

[0149] Compound l is a potent and selective small molecule inhibitor of certain mutated forms of the isocitrate dehydrogenase 1 (IDH-1) enzyme. Compound 1 selectively inhibits mutant IDH-1 enzymes compared to the wild type IDH-1 enzyme, targeting the mutant IDH-1 variants defined herein as R132X mIDH-1 Mutation(s). Example 1 provides in vitro data demonstrating inhibition of various R132X mutations of mIDH-1 enzyme. For example, Compound 1 targets the mutant IDH-1 variants R132H, R132C, R132L, R132G, and R132S using assays described in Example 1 with IC50 concentrations that are approximately at least 180-fold lower than the wild-type IDH-1 enzyme in vitro. In addition, Compound 1 targets the R132H and R132C mutations of IDH-1 at IC50 concentrations demonstrating selectivity over wild-type IDH-1 enzyme in vitro (based on IC50 measurements as determined in Example 1 based on average +/- SEM, nM). Accordingly, preferred R132X mutations include R132H and R132C, as well as R132L, R132G, and R132S (or other R132X mutations having therapeutically relevant 2-HG IC50 values obtained using the in vitro assay of Example 1). In addition, Compound 1 selectively inhibits mutant IDH-1 compared to mutant IDH-2 forms. The selectivity of Compound 1 against other IDH isozymes was tested using diaphorase coupled assays employing either wild-type IDH-1 or one of 2 alternative mutated forms of IDH-2 (R140Q and R172K). Compound 1 had very weak activity against either wild type IDH-1 or R172K IDH-2 mutation (with enzymatic IC50 measurements obtained according to Example 1 of about 20-25 micromolar, compared with IC50 values of less than about 150 nanomolar obtained for the R132X m-IDH-1 Mutations). In addition, Compound 1 did not show any inhibition of R140Q IDH-2 up to a concentration of 100 micromolar. These selectivity data demonstrate that Compound 1 is a potent and selective inhibitor of enzymes harboring R132X mIDH-1 Mutation(s).

[0150] The R132X mIDH-1 Selective Inhibitor Therapy (single agent or combination) can be administered to adult patients with an IDH-1 mutation as detected by a medically appropriate (e.g., FDA-approved) test for mIDH-1 mutation(s). Preferably, the test is a diagnostic that identifies an R132X mIDH-1 Mutation(s) in the patient prior to the administration of Compound 1. Preferably, the patient is identified as harboring one or more R132X mIDH-1 Mutation(s) based on Next Generation Sequencing (NGS) detection on a tissue sample obtained from the patient prior to the administration of Compound 1 and/or administration of any R132X mIDH-1 Selective Inhibitor Therapy.

[0151] A patient in need of R132X mIDH-1 Selective Inhibitor Therapy can have an elevated level of 2-HG measured in the patient (e.g., in the blood plasma of the patient) prior to initiating any R132X mIDH-1 Selective Inhibitor Therapy. Preferably, the level of 2-HG measured in the blood of the patient declines during the first 2 weeks of the Course of Treatment of a R132X mIDH-1 Selective Inhibitor Therapy. For example, a patient may have a measured blood concentration level of 2-HG that is greater than about 200 ng/mL of 2-HG prior to the administration of Compound 1 pursuant to administration of a R132X mIDH-1 Selective Inhibitor Therapy to the patient in need thereof, and a measured blood concentration level of 2- HG of less than about 200 ng/mL during a Course of Treatment with a R132X mIDH-1 Selective Inhibitor Therapy. For example, in the human clinical trial disclosed in Example 11, all IDH- lm+ patients had elevated 2-HG, which was reduced upon treatment with Compound 1 by day 15 of the Course of Treatment, with about 30% demonstrating a response to the administration of Compound 1 at some point during the Course of Treatment. In this patient population, the normal 2-HG measured in patient blood at CRL was about 70 ± 17 ng/mL; the observed highest was about 91 ng/mL and the lowest was about 43 ng/mL. Preferably, the R132X mIDH-1 Selective Inhibitor Therapy consists of the administration of Compound 1 (i.e., Compound 1 is the only mIDH-1 inhibitor administered to the patient throughout the Course of Treatment).

[0152] Compound 1 is administered over a therapeutically effective Course of Treatment, which is preferably long enough to provide and sustain an intended therapeutic effect. For example, the Course of Treatment can be long enough to therapeutically reduce elevated 2-HG levels in a patient (e.g., reduce 2-HG levels measured in patient blood plasma to below about 200 ng/mL), with continued administration of Compound 1 to the patient in a manner that provides therapeutically effective steady state blood plasma concentration levels of Compound 1 (e.g., trough blood plasma concentrations greater than the IC90 concentration value for 2HG production measured for a R132X IDH-1 mutation identified in cells obtained from the patient). When treating patients with elevated 2-HG levels measured in the patient’s blood prior to administration of Compound 1, the Course of Treatment can be at least a number of consecutive days starting from the initial administration of Compound 1 to the patient with elevated 2-HG levels, and continuing with daily administration of Compound 1 (e.g., 150 mg BID) for at least a number of days effective to reduce the 2-HG levels measured in the blood of the patient to less than about 200 ng/mL (preferably less than 180 ng/mL) and/or a level considered medically appropriate for the patient (e.g., to a range determined to be medically normal for that patient in the treatment paradigm). Preferably, the Course of Treatment is at least 15 consecutive days starting with the day of the initial administration of Compound 1 to the patient and comprises 150 mg of the solid oral dosage form of Compound 1 (e.g., Example 5) administered to the patient twice per day (e.g., every 12 hours) every day for at least 15 days. The Course of Treatment can be one or more 28-day treatment cycles of daily BID administration of 150 mg of Compound 1 in the solid oral dosage form obtained from Example 5. The Course of Treatment can continue throughout a medically appropriate number of days for a patient. For example, the Course of Treatment can last for any medically appropriate number of consecutive 28-day treatment cycles, including a Course of Treatment lasting for 1, 2, 3, 4, 5, 6 consecutive treatment cycles of 28-days each, and/or a Course of Treatment of 20 weeks, and/or a Course of Treatment of 6 months or more. In some methods, the Course of Treatment is at least 6 months, or between at least 15 consecutive days and 6 months of consecutive days of treatment comprising administration of 150 mg BID of Compound 1 in a pharmaceutical form obtained from Example 5.

[0153] The R132X mIDH-1 Selective Inhibitor Therapy (e.g., oral administration of 150 mg BID of Compound 1 throughout a Course of Treatment) provided unexpectedly durable blood concentration levels throughout the Course of Treatment exceeding 6 months. The level of steady state blood concentration during the Course of Treatment was durable, meaning that the steady state blood concentration of Compound 1 did not decrease by more than 10% throughout a Course of Treatment with continued administration of Compound 1 at a dose of 150 mg BID each day, while remaining within a therapeutic concentration window defined by a minimum concentration above the IC90 determined in vitro for the 2-HG production of a R132X mIDH-1 mutation harbored by the patient (e.g., above about 2,000 ng/mL or above about 1652 ng/mL), and a maximum concentration value below the concentration associated with medically unacceptable risk of QTc prolongation (e.g., about 7,200 ng/mL or about 7840 ng/mL).

[0154] In addition, the R132X mIDH-1 Selective Inhibitor Therapy can reduce 2-HG levels in the blood of the patient, although this reduction did not correlate with disease response in the patients during the Course of Treatment. As shown in Figures 19A-19D, the R132X mIDH-1 Selective Inhibitor Therapy provided a sustained reduction in 2HG levels, meaning that elevated levels of the blood concentration of 2-HG in patient blood plasma were reduced relative to pre dose levels during the initial portion of a Course of Treatment (e.g., within the first 15 days) and then sustained at or below about the pre-dose level throughout the rest of the Course of

Treatment (e.g., at or below about 200 ng/mL for 6 consecutive 28-day treatment cycles or at or below about 180 ng/mL for 6 consecutive 28-day treatment cycles).

[0155] Figures 18A-18E is a graph showing the steady state (trough) blood concentration measured in patients after administration of 150 mg of Compound 1 BID, as described in the human clinical trial of Example 11. Figures 18A-18E are different graphs each showing the steady state blood concentration of Compound 1 measured in patient blood for a collection of patients in the human clinical trial described in Example 11. Figures 18A-18E are each graphs showing the measured steady state blood concentrations measured in each of individual patients included in the population, at the indicated time points, in the human clinical trial of Example 11. Notably, the steady state blood concentration of Compound 1 remained above a therapeutic minimum value (e.g., the IC90 value for 2-HG production by at least one of the R132X mutation(s) identified in patient’s tissue prior to administration of Compound 1) throughout the Course of Treatment (e.g., 30 weeks). In particular, the administration of Compound 1 in the oral dosage form described in Example 5 at 150 mg BID resulted in a sustained therapeutically effective trough blood plasma concentration above 2,000 ng/mL throughout a 30-week Course of Treatment as shown in Figures 18A-18E (e.g., including after cycle 3 of a 28-day treatment cycle, after week 20 and continuing to week 30). The trough concentration measurements of Compound 1 in patient blood did not decline below a therapeutically effective level at each point measured during the Course of Treatment. Referring to Figures 19A-19D, the administration of Compound 1 in the oral dosage form from Example 5 at 150 mg BID in the human clinical trial of Example 11 resulted in a sustained 2-HG level under 200 ng/mL in plasma throughout the Course of Treatment ranging from cycle 2, day 1 through cycle 8, day 1 (each cycle represents 28 consecutive days of oral administration of 150 mg BID of the solid form of Compound 1 obtainable from the production method of Example 5).

[0156] This disclosure is based in part on the discovery that a R132X mIDH-1 Selective Inhibitor Therapy where Compound 1 is the only mIDH-1 inhibitor administered (administration of Compound 1 at 150 mg BID) unexpectedly resulted in a steady state blood concentration that was durable (e.g., blood plasma steady state concentration of Compound 1 remains within about 20% (or does not decrease by more than about 20%), and preferably remains within 10% (or does not decrease by more than 10%) of the concentration measured the day after the initial 28- day cycle in a Course of Treatment). In addition, the administration of Compound 1 as described in Example 11 reduced elevated 2-HG concentrations in the blood of the patients within about 15 days and then sustained patient blood concentrations of 2-HG at less than about 200 ng/mL after about 15 days of a Course of Treatment.

[0157] The R132X mIDH-1 Selective Inhibitor Therapy can provide a sustained ratio of greater than about 10 (preferably greater than about 20) of blood plasma concentration of Compound 1 (e.g., trough concentrations measured pre-dose at concentrations of about 2,000 ng/mL or greater) to 2-HG blood level (e.g, concentrations of plasma concentrations of about 200 ng/mL or lower, including concentrations of about 100 ng/mL) after cycle 3 day 1 (i.e., after BID doses administered over the initial 15 consecutive days of treatment) of a 28-day treatment cycle. A plasma half-life of about 60 hours was estimated for Compound 1, with steady state achieved by week 2 of the course of treatment. The steady state blood concentrations of Compound 1 measured in the patients was above the IC90 value for 2-HG inhibition in R132X mIDH-1 cells (described in the Examples). As shown in Figures 18A-18E, the plasma exposures (steady state blood plasma concentration) of Compound 1 were durable (i.e., sustained) throughout the 30-week treatment duration. As shown in Figures 19A-19D, the plasma 2-HG concentrations were reduced to the normal range within 1 cycle (C2D1) and maintained throughout the treatment duration. No dose limiting toxicities of Compound 1 were observed during dose escalation studies, and the maximum tolerated dose (MTD) of Compound 1 was not reached. Preferably, the R132X mIDH-1 Selective Inhibitor Therapy can be administered to a patient that has not received any other R132X mIDH-1 inhibitor compound.

[0158] As described in Example 11, Figure 21 A shows the ratio of the steady state blood concentration of Compound 1, normalized to 1.0 using the concentration measured on day 15 of a Course of Treatment, for a single patient who received 150 mg BID of the solid form of Compound 1 obtainable from Example 5 throughout a Course of Treatment of over 300 days (i.e., greater than 6 months). The steady state blood exposure (concentration) of Compound 1 varied from about 90-133% of the concentration of Compound 1 measured in the patient at cycle 1, day 15, throughout the subsequent remainder of this Course of Treatment.

[0159] Optionally, a hypomethylating agent and/or a nucleic acid synthesis inhibitor can also be administered to the patient during the Course of Treatment. Suitable agents that can also be administered during the Course of Treatment include azacitabine and/or decitabine.

[0160] In some embodiments, a combination therapy of Compound 1 and azacitidine can be administered for the treatment of patients with certain forms of cancer (e.g., glioma) harboring IDH-1 mutations. For example, patients can be administered Compound 1 daily (BID) in continuous 28-day cycles, in combination with azacitidine (administered at the dose of 75mg/m2 for 7 days IV/SC per every 28-day cycle). Azacitidine (also, azacytidine or AZA herein) is believed to exert its antineoplastic effects by causing hypomethylation of DNA and direct cytotoxicity on abnormal hematopoietic cells in the bone marrow. Azacitidine can be administered during a Course of Treatment at a subcutaneous dose of 75 mg/m2 daily for 7 days every 4 weeks. The azacitidine dose can be increased to 100 mg/m2 if no beneficial effect was seen after 2 treatment cycles. The dose of azacitidine can be decreased and/or delayed based on hematologic response or evidence of renal toxicity. Azacitidine is indicated for treatment of patients with the following myelodysplastic syndrome subtypes: refractory anemia or refractory anemia with ringed sideroblasts (if accompanied by neutropenia or thrombocytopenia or requiring transfusions), refractory anemia with excess blasts, refractory anemia with excess blasts in transformation, and chronic myelomonocytic leukemia. In some embodiments, a method of treatment comprises (a) the (e.g., oral) administration of a total of 150 mg of

Compound 1 BID to a patient throughout a Course of Treatment; and (b) administering a therapeutically effective amount of azacitidine to the patient (e.g, administering azacitidine at a dose of 75 mg/m2 daily for 7 days every 4 weeks, wherein the azacitidine dose can be increased to 100 mg/m2 if no beneficial effect was seen after 2 treatment cycles and the dose of azacitidine can be decreased and/or delayed based on hematologic response or evidence of renal toxicity).

[0161] Decacitabine (5-aza-2'-deoxycytidine) is a nucleoside metabolic inhibitor indicated for treatment of patients with myelodysplastic syndromes (MDS) including previously treated and untreated, de novo and secondary MDS of all French-American-British subtypes (refractory anemia, refractory anemia with ringed sideroblasts, refractory anemia with excess blasts, refractory anemia with excess blasts in transformation, and chronic myelomonocytic leukemia) and intermediate- 1, intermediate-2, and high-risk International Prognostic Scoring System groups. In some embodiments, a method of treatment comprises the (e.g., oral) administration of a total of 150 mg of Compound 1 BID to a patient throughout a Course of Treatment; and (b) administering a therapeutically effective amount of decacitabine to the patient (e.g, administering decacitabine at a dose of 15 mg/m2 by continuous intravenous infusion over 3 hours repeated every 8 hours for 3 days and repeating this cycle every 6 weeks; or administering the

decacitabine at a dose of 20 mg/m2 by continuous intravenous infusion over 1 hour repeated daily for 5 days. Repeat cycle every 4 weeks).

Glioma

[0162] The present disclosure also provides methods for treating solid tumors in the CNS, including a brain cancer tumor, harboring a R132 IDH-1 mutation. For example, patients diagnosed with brain cancer harboring a mutant IDH-1 cancer cell can be treated with a therapeutically effective amount of Compound 1 in combination with azacitidine.

[0163] Compound l is a small molecule inhibitor of mutated forms of isocitrate

dehydrogenase 1 (IDH-1) enzyme, and is useful for the treatment of adult patients diagnosed with cancer having an IDH-1 mutation as detected by an FDA-approved test. Compound 1 can be administered to patients in need thereof in a therapeutically effective amount (e.g., 150 mg orally twice daily until disease progression or unacceptable toxicity). Patients for the treatment of cancer with Compound 1 can be selected based on the presence of IDH-1 mutations in the blood or bone marrow. In one embodiment, the recommended starting dose of Compound 1 is 150 mg taken orally twice daily with or without food until disease progression or unacceptable toxicity. For patients without disease progression or unacceptable toxicity, the patient can receive the therapeutically effective amount of Compound 1 for a minimum of 6 months to allow time for clinical response.

[0164] The disclosure is based in part on preclinical studies showing that Compound 1 can cross the blood brain barrier (BBB) in mouse models. Oral administration of Compound 1 showed high systemic bioavailabilty in multiple preclinical species. Permeability was excellent, with little evidence of efflux, and significant brain penetration was observed in mice (98% brain binding in murine animal model).

[0165] Preferably, patients diagnosed with glioma harboring a R132 IDH-1 mutation can be treated with a therapeutically effective combination of a pharmaceutical composition of

Compound 1 (e.g., an oral dosage form providing 150 mg of Compound 1 administered twice per day on consecutive days for a course of treatment comprising multiple treatment cycles totaling at least 6 months) and azacitidine. The azacitidine can be subcutaneously or intravenously administered to the patient in an azacitidine treatment cycle consisting of the administration of a total dose of 75 mg/m2 each day for 7 consecutive days beginning at the start of each treatment cycle, followed by 21 consecutive days without administration of the azacitidine to the patient.

A 48-hour dose-interruption of azacitidine is allowed for weekends or holidays. If no response is seen after 2 treatment cycles, azacitidine can be administered at a total dose of 100 mg/m2 each day.

[0166] Compound 1 is preferably administered on consecutive days throughout a Course of Treatment. For example, a Course of Treatment can comprise one or more 28-day treatment cycles. Preferably, the Course of Treatment is at least about 15 consecutive days, at least about a 28-consecutive day treatment cycle, or at least about four, five, six or more consecutive 28-day treatment cycles and more preferably, at least about 4 months or longer (e.g., 6 months or longer). Preferably, Compound 1 is administered twice per day (e.g., about every 12 hours) every day throughout a Course of Treatment. [0167] In some embodiments, patients can be treated with Compound 1 in combination with a hypomethylating agent such as azacitidine or decitabine. The recommended starting dose for azacitidine in the first treatment cycle, for all patients regardless of baseline hematology laboratory values, is 75 mg/m2 of body surface area, injected subcutaneously, daily for 7 days, followed by a rest period of 21 days (28-day treatment cycle). It is recommended that patients be treated for a minimum of 6 cycles. Treatment should be continued as long as the patient continues to benefit or until disease progression. In some methods, azacitidine is administered to the patient in need thereof at a dose of 75 mg/m2, SC, dl-7, q4 wk throughout a course of treatment, while receiving Compound 1 at a dose of 150 mg BID. In other methods, decitabine is administered to the patient in need thereof at a dose of 20 mg/m2, IV, dl-5, q4 wk, while receiving Compound 1 at a dose of 150 mg BID.

[0168] In one embodiment, patients diagnosed with glioma harboring a mIDHI can be treated with a mIDHI Inhibitor Therapy consisting of Compound 1 and azacitidine. Treatment with the hypomethylating agent azacitidine can cause tumor growth inhibition in a patient- derived IDHl -mutated glioma model by reducing DNA methylation and inducing glial differentiation. IDHl R132H mutations represent more than 90% of the IDH mutations present in low grade glioma and secondary GBM patients. The IDHl mutations R132C and R132S are also reported in glioma patients. At least in mIDHI harboring cancer cells, wild type and mutant IDHl form a heterodimeric complex that can produce very high 2-HG levels (up to 3-35 mM in glioma cells). For example, patients bearing IDHl mutations have elevated levels of 2-HG, which in some cases reach tumor concentrations >10 mM (glioma).

[0169] In another embodiment, patients diagnosed with chondrosarcoma harboring a mutant IDHl cancer cell can be treated with a therapeutically effective amount of Compound 1 alone or in combination with azacitidine. In some embodiments, a combination therapy comprising Compound 1 and azacitidine can be administered for the treatment of patients with

chondrosarcoma harboring IDHl mutations. For example, patients can be administered

Compound 1 daily (BID) in continuous 28-day cycles, in combination with azacitidine

(administered at the daily dose of 75 mg/m2 for 7 days IV/SC per every 28-day cycle).

[0170] Preferably, patients treated with a combination of Compound 1 and azacitidine receive a therapeutically effective amount of a mIDHI Inhibitor Therapy selected from a dose level indicated in Table 3 below. Table 3 - Preferred Dose Levels for mIDHI Inhibitor Therapy with Azacitidine

[0171] Patients diagnosed with hepatobiliary carcinoma (HBC) harboring a mutant IDH1 cancer cell can be treated with a therapeutically effective amount of Compound 1 alone or in combination with a PD-1 inhibitor (e.g., Pembrolizumab (Keytruda) or Nivolumab (Opdivo)). In some embodiments, a combination therapy of Compound 1 and the PD-1 inhibitor can be administered for the treatment of patients with a HBC cancer harboring IDH1 mutations. For example, patients can be administered compound 1 daily (BID) in continuous 28-day cycles, in combination with Pembrolizumab (e.g., administered at the dose of 200 mg every 3 weeks). For example, patients can be administered compound 1 daily (BID) in continuous 28-day cycles, in combination with Nivolumab (e.g., administered at the dose of 240 mg every 2 weeks or 480 mg every 4 weeks). Preferably, patients treated with a combination comprising Compound 1 and Nivolumab receive a therapeutically effective amount of a mIDHI Inhibitor Therapy selected from a dose level indicated in Table 4 below.

Table 4 - Preferred Dose Levels for mIDHI Inhibitor Therapy with Nivolumab

[0172] Patients diagnosed with IHCC harboring a mutant IDH1 cancer cell can be treated with a therapeutically effective amount of Compound 1 alone or in combination with a chemotherapy (e.g., gemcitabine and cisplatin). Preferably, patients treated with a combination of Compound 1 and gemcitabine and cisplatin chemotherapy receive a therapeutically effective amount of a mIDHI Inhibitor Therapy selected from a dose level indicated in Table 5 below.

Table 5 - Preferred Dose Levels for mIDHI Inhibitor Therapy with Gemcitabine/Cisplatin

Diagnostic Methods

[0173] Patients can be selected for treatment according to methods described herein using various diagnostic technologies. A patient can be identified as having a R132X mutation in mIDHI using a diagnostic method disclosed herein prior to the administration of Compound 1 to the patient. The R132X gene mutation can be determined prior to administration of Compound 1 to the patient. Compound 1 can be administered to patients who have received prior anticancer therapy or other concomitant (non-anticancer) medications. In some examples, Compound 1 is administered to patient who has not received a prior mIDH-1 inhibitor therapy.

[0174] In some embodiments, the present disclosure provides methods of treating acute myeloid leukemia (AML) in patients with an isocitrate dehydrogenase- 1 (IDH1) mutation, the method comprising steps of:

a. providing DNA from a sample obtained from a patient;

b. detecting an IDHl mutation in the DNA from the sample; and

c. administering to the patient with the IDHl mutation a total of 150 mg of Compound 1 twice daily in a pharmaceutically acceptable composition.

[0175] In some embodiments, the present disclosure provides methods of treating acute myeloid leukemia (AML) in patients with an isocitrate dehydrogenase- 1 (IDHl) mutation, the method comprising steps of:

a. isolating and purifying DNA from a sample obtained from a patient;

b. detecting an IDHl mutation in the DNA from the sample; and c. administering to the patient with the IDH1 mutation a total of 150 mg of Compound 1 twice daily in a pharmaceutically acceptable composition.

[0176] In some embodiments, the present disclosure provides methods of treating AML in patients with an isocitrate dehydrogenase- 1 (IDH1) mutation, the method comprising steps of: determining whether the patient has an IDHl mutation by:

i. obtaining a sample from the patient; and

ii. performing an assay (e.g., an FDA-approved diagnostic test, such as the IDHl Assay of Example 14) on the sample to determine if the patient has an IDHl mutation; and

if the patient has an IDHl mutation, then administering to the patient with the IDHl mutation a total of 150 mg of Compound 1 twice daily in a pharmaceutically acceptable composition, and

if the patient does not have an IDHl mutation, then not administering to the patient with the IDHl mutation a total of 150 mg of Compound 1 twice daily in a

pharmaceutically acceptable composition.

[0177] The present disclosure also provides methods of treating acute myeloid leukemia (AML) in patients with an isocitrate dehydrogenase- 1 (IDHl) mutation, the method comprising administering twice daily to a patient with an IDHl mutation 150 mg of Compound 1 in a pharmaceutically acceptable composition, wherein the IDHl mutation has been detected using an FDA-approved diagnostic test.

[0178] In some embodiments, the present disclosure provides methods of treating glioma in patients with an isocitrate dehydrogenase- 1 (IDHl) mutation, the method comprising steps of: a. providing DNA from a sample obtained from a patient;

b. detecting an IDHl mutation in the DNA from the sample; and

c. administering to the patient with the IDHl mutation a total of 150 mg of Compound 1 twice daily in a pharmaceutically acceptable composition.

[0179] In some embodiments, the present disclosure provides methods of treating glioma in patients with an isocitrate dehydrogenase- 1 (IDHl) mutation, the method comprising steps of: a. isolating and purifying DNA from a sample obtained from a patient;

b. detecting an IDHl mutation in the DNA from the sample; and

c. administering to the patient with the IDHl mutation a total of 150 mg of Compound 1 twice daily in a pharmaceutically acceptable composition. [0180] In some embodiments, the present disclosure provides methods of treating glioma in patients with an isocitrate dehydrogenase- 1 (IDH1) mutation, the method comprising steps of: determining whether the patient has an IDHl mutation by:

i. obtaining a sample from the patient; and

ii. performing an assay (e.g., an FDA-approved diagnostic test, such as the IDHl Assay of Example 14) on the sample to determine if the patient has an IDHl mutation; and

if the patient has an IDHl mutation, then administering to the patient with the IDHl mutation a total of 150 mg of Compound 1 twice daily in a pharmaceutically acceptable composition, and

if the patient does not have an IDHl mutation, then not administering to the patient with the IDHl mutation a total of 150 mg of Compound 1 twice daily in a

pharmaceutically acceptable composition.

[0181] The present disclosure also provides methods of treating glioma in patients with an isocitrate dehydrogenase- 1 (IDHl) mutation, the method comprising administering twice daily to a patient with an IDHl mutation 150 mg of Compound 1 in a pharmaceutically acceptable composition, wherein the IDHl mutation has been detected using an FDA-approved diagnostic test.

[0182] In some embodiments, the IDHl mutation is an IDHl R132 mutation. Examples of an IDHl R132 mutation include R132C, R132H, R132S, R132G, and R132L. In some embodiments, the IDHl R132 mutation is R132C. In some embodiments, the IDHl R132 mutation is R132H. In some embodiments, the IDHl R132 mutation is R132S. In some embodiments, the IDHl R132 mutation is R132G. In some embodiments, the IDHl R132 mutation is R132L. In some embodiments, the patient is harboring an IDHl mutation, such as an IDHl R132 mutation selected from the group consisting of R132C, R132H, R132S, R132G, and R132L.

[0183] In some embodiments, the AML is relapsed or refractory AML.

[0184] In some embodiments, the patient is receiving or has received anticancer therapy. In some embodiments, the patient is resistant or refractory to prior anticancer therapy. In some embodiments, the patient is receiving or has received therapy comprising azacitidine or cytarabine. [0185] Compound 1 can be administered as provided herein. For example, Compound 1 can be administered as an oral dosage form, such as a tablet or a capsule. For example, Compound 1 can be administered as part of a combination therapy comprising, e.g., Compound 1 and azacitidine or cytarabine. For example, Compound 1 can be administered as the PRODUCT described in Example 5.

[0186] In some embodiments, provided methods further comprise not administering

Compound 1 if the patient does not have an IDH1 mutation, as determined, e.g., by an FDA- approved diagnostic test.

[0187] In some embodiments, provided methods comprise detecting an IDHl mutation and administering Compound 1 as described herein. In some embodiments, IDHl mutations can be detected using an FDA-approved diagnostic test, such as the IDHl Assay described in Example 14.

[0188] In some embodiments, detecting an IDHl mutation comprises detecting a single nucleotide variant (SNV) coding the IDHl mutation. In some embodiments, the IDHl mutation is selected from the group consisting of R132C, R132H, R132G, R132S, and R132L. In some embodiments, detecting an IDHl R132C mutation comprises detecting the SNV: TGT. In some embodiments, detecting an IDHl R132H mutation comprises detecting the SNV: CAT. In some embodiments, detecting an IDHl R132G mutation comprises detecting the SNV: GGT. In some embodiments, detecting an IDHl R132S mutation comprises detecting the SNV: AGT. In some embodiments, detecting an IDHl R132L mutation comprises detecting the SNV: CTT.

[0189] In some embodiments, the IDHl mutation is detected using PCR technology with homogenous real-time fluorescence detection. In some embodiments, the IDHl mutation is detected using an in vitro polymerase chain reaction (PCR) assay for the qualitative detection of single nucleotide variants (SNVs) coding an IDHl R132 mutation selected from the group consisting of R132C, R132H, R132G, R132S, and R132L in the DNA from a sample.

[0190] In some embodiments, the diagnostic test uses a sample obtained from the patient. In some embodiments, the sample is a blood or tissue sample. In some embodiments, the sample is patient bone marrow. In some embodiments, the sample is patient blood. In some embodiments, the sample has been preserved with EDTA.

[0191] In some embodiments, provided methods further comprise: lysing cells from the sample at an elevated temperature in a lysis buffer comprising guanidine isothiocyanate;

capturing DNA released from the lysed cells using magnetic microparticles;

washing the captured DNA; and

eluting the captured DNA from the magnetic microparticles with elution buffer to give an extracted DNA sample.

[0192] In some embodiments, provided methods further comprise:

combining the extracted DNA sample, a DNA polymerase, oligonucleotide primers,

deoxyribonucleoside triphosphates (dNTPs), and magnesium chloride in a well of a 96- well plate to give a mixture;

sealing the 96-well plate with a cover;

activating the DNA polymerase at a high temperature;

subjecting the mixture to thermal cycling comprising multiple rounds of heating (e.g., to a high temperature suitable to melt double-stranded DNA) and cooling (e.g., to a low temperature suitable to promote annealing of primers to their respective targets); and measuring the real-time fluorescence signals of the mixture.

[0193] In some embodiments, the oligonucleotide primers are designed to specifically amplify (i) R132C and R132H mutations or (ii) R132G, R132S, and R132L mutations. In some embodiments, the oligonucleotide primers are designed to specifically amplify R132C and R132H mutations. In some embodiments, the oligonucleotide primers are designed to specifically amplify R132G, R132S, and R132L mutations. In some embodiments, two samples from the same patient are evaluated, so that one sample is mixed with oligonucleotide primers that are designed to specifically amplify R132C and R132H mutations and the other sample is mixed with oligonucleotide primers that are designed to specifically amplify R132G, R132S, and R132L mutations.

[0194] In some embodiments, the DNA polymerase is a thermophilic enzyme that has been chemically modified to render it inactive at ambient temperature.

[0195] In some embodiments, provided methods comprise an internal control. For example, in some embodiments, the mixture further comprises oligonucleotide primers designed to amplify a region of the IDH1 gene outside of codon 132, thereby serving as an endogenous internal control. [0196] In some embodiments, the real-time fluorescence signal of each IDHl mutation of either (i) R132C and R132H or (ii) R132G, R132S, and R132L is distinguishable in a single well. In some embodiments, the real-time fluorescence signal of the internal control and each IDHl mutation of either (i) R132C and R132H or (ii) R132G, R132S, and R132L is

distinguishable in a single well.

[0197] Without wishing to be bound by any particular theory, it may be desirable for a diagnostic test described herein to be performed in such a way as to prevent nucleic acid contamination. In some embodiments, the diagnostic test is performed in a sealed 96-well plate. In some embodiments, the diagnostic test is performed without opening the sealed 96-well plate. In some embodiments, aerosol barrier pipette tips are used for all pipetting in provided methods. In some embodiments, the diagnostic test is performed in a dedicated area.

[0198] In some embodiments, provided methods further comprise detecting a co-mutation described herein (e.g., using a method of detection as described herein).

[0199] Compound /Referring to Figure 5, Compound 1 can be prepared in a convergent synthesis from Intermediate A and Intermediate B as shown in Figure 5 via the nucleophilic displacement reaction under basic conditions of (S)-3-(l-aminoethyl)-6-chloroquinolin-2(lH)- one (Intermediate A) and the fluoropyridone (Intermediate B). 1H, 13C NMR and mass spectral data are consistent with the assigned structure. The asymmetric synthesis of Intermediate A started with the condensation of the commercially available quinoline aldehyde (1) with (R)-tert- butanesulfmamide (2) to form the chiral (R)-N-tert-butanesulfmimine (3), followed by addition of methyl magnesium bromide in di chi orom ethane to yield the desired product (4) as the major diastereoisomer (dr: 98:2). Cleavage of the chiral auxiliary and simultaneous hydrolysis of 2- chloroquinoline moiety under mildly acidic conditions using IN HC1 in dioxane gave

Intermediate A in quantitative yield. The structure of Intermediate A was confirmed by NMR and mass spectroscopy, and the enantiomeric purity was determined by chiral SFC analysis. The (S)-stereochemistry was confirmed by X-ray co-crystal structures of several inhibitor analogs prepared from the same chiral amine intermediate bound to mIDH-1 R132H. Intermediate (B) was prepared from commercially available 5-fluoropicolinonitrile in four steps. N-oxidation of 5- fluoropicolinonitrile followed by reflux of the N-oxide in acetic anhydride gave the acetate, following work-up and purification. Solvolysis of the acetate group followed by N-methylation under standard conditions gave a mixture of N-m ethylated and O-methylated products (4: 1). The minor O-methylated product was removed by column chromatography. NMR and mass spectral data are consistent with the structure of Intermediate Compound (B). Compound 1 (5-{[(l S)-l- (6-chloro-2-oxo- 1 ,2-dihydroquinolin-3 -yl)ethyl]amino} - 1 -methyl-6-oxo- 1 ,6-dihydropyridine-2- carbonitrile) has a molecular weight of 355 with a melting point onset temperature of 251.3 °C (DSC) and peak maximum 254.1 °C.

[0200] Compound 1 is also known as olutasidenib. Compound 1 can also be identified by the following chemical names:

• 2-Pyridinecarbonitrile, 5-[[(lA)-l-(6-chloro-l,2-dihydro-2-oxo-3- quinolinyl)ethyl]amino]- 1 ,6-dihydro- 1 -methyl-6-oxo-;

• 5 - { [( 1 A)- 1 -(6-chloro-2-oxo- 1 ,2-dihy droquinolin-3 -yl)ethyl] amino } - 1 -methyl-6-oxo- 1,6- dihydropyridine-2-carbonitrile; and

• (S)- 5-(( 1 -(6-chloro-2-oxo- 1 ,2-dihy droquinolin-3 -yl)ethyl)amino)- 1 -methyl-6-oxo- 1,6- dihydropyridine-2-carbonitrile.

[0201] Compound 1 also has the following identifiers:

• Code designation: FT-2102

• CAS Registry Number: 1887014-12-1

• UNII: 0T4IMT8S5Z

• WHO Number: 11036

[0202] An oral unit dosage form comprising a pharmaceutically acceptable solid form of Compound 1 (e.g., as obtained from Example 5) can be formulated as a drug product with various inactive components as excipients (e.g., as a tablet or capsule) (referred to as the “PRODUCT”). Each drug product excipient in PRODUCT meets the requirements of the respective current United States Pharmacopeia (USP) or National Formulary (NF) monograph. The capsule shells can comprise gelatin and about 2.9% w/w of titanium dioxide (E171).

Preferably, each oral unit dosage form comprises a total of 50 mg or 150 mg of the

pharmaceutically acceptable form of Compound 1 (e.g., micronized crystalline olutasidenib) combined (e.g., at 30-50% w/w) as the only active moiety with pharmaceutically acceptable excipients such as a filler (e.g., AVICEL PH101 @50 micron, AVICEL PH102 @100 micron), a disintegrant (e.g., Ac-Di-Sol), optionally one or more compounds as a lubricant (e.g., magnesium stearate), a glidant/anti-adherent, and/or anti-static (e.g., colloidal silicon dioxide). The excipients can form about 50-70% by weight of the pharmaceutical oral unit dosage form. In one example, a capsule or tablet comprises a total of about 33% of Compound 1, with the remaining weight of the capsule or tablet is formed from excipients and/or capsule material (e.g., a gelatin). Alternatively, the PRODUCT can be provide as tablet for oral administration. Each tablet can contain the following inactive ingredients: colloidal silicon dioxide, croscarmellose sodium, hypromellose acetate succinate, magnesium stearate, microcrystalline cellulose, and sodium lauryl sulfate. The tablet coating can include FD&C blue #2, hypromellose, lactose monohydrate, titanium dioxide, and/or triacetin.

[0203] It will be appreciated that use of headers in the present disclosure are provided for the convenience of the reader. The presence and/or placement of a header is not intended to limit the scope of the subject matter described herein.

Exemplary Embodiments

[0204] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient diagnosed with a cancer harboring a cancer cell with an IDH-1 R132 mutation selected from the group consisting of: R132L, R132G, and R132S, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1.

2. The method of embodiment 1, wherein the cancer does not harbor a IDH-2 mutation.

3. The method of embodiment 1, wherein the cancer does not harbor a IDH-2 mutation selected from the group consisting of: IDH-2 R172K and IDH-2 R140Q.

4. The method of embodiment 1, wherein the patient is diagnosed as having a R132

mutation based on a patient diagnostic.

5. The method of embodiment 4, wherein the patient diagnostic comprises detecting the R132 mutation in a tissue sample obtained from the patient.

6. The method of embodiment 5, wherein the tissue sample is obtained from the bone

marrow of the patient.

7. The method of any one of embodiments 4-6, wherein the R132 mutation is detected using next generation sequencing (NGS) without the use of PCR.

8. A method of treatment comprising the steps of: a. selecting a patient for treatment based on the presence of one or more IDH-1 mutations selected from the group consisting of: R132L, R132G, and R132S; b. administering Compound 1 to the selected patient from step (a) at a starting dose of 150 mg taken orally twice daily until disease progression or unacceptable toxicity.

The method of embodiment 8, where the IDH-1 mutation is detected in cancer cells obtained from the blood or bone marrow of the patient.

The method of embodiment 9, wherein the IDH-1 mutation is detected prior to administering Compound 1 to the patient.

The method of any one of embodiments 1-10, comprising the step of detecting the IDH-1 mutation in a cell from the patient using a next-generation sequencing (NGS)-based tumor genotyping assay.

The method of any one of embodiments 1-11, wherein administration of Compound 1 to the patient results in a decreased 2-hydroxyglutarate (2-HG) levels in the blood of the patient within the first 15 consecutive days of treatment of the patient with Compound 1. The method of any one of embodiments 1-12, wherein the method comprises

administering 150 mg of Compound 1 to the patient in the solid form obtained from the method of Example 5.

The method of any one of embodiments 1-12, wherein the method comprises

administering 150 mg of Compound 1 to the patient twice daily throughout a course of treatment.

The method of embodiment 14, wherein the course of treatment is at least 15 consecutive days.

The method of any one of embodiments 1-15, wherein Compound 1 is administered to the patient once every 12 hours on consecutive days throughout a course of treatment. The method of any one of embodiments 1-16, wherein Compound 1 is administered to the patient throughout a course of treatment of at least 6 months.

A method of inhibiting the production of 2-HG from a cell harboring a IHD-1 mutation selected from the group consisting of: R132L, R132G and R132S, the method comprising contacting the cell with Compound 1 in an amount, under conditions, and for a time sufficient to inhibit the production of 2-HG from the cell. A method of treating a patient diagnosed with a cancer harboring a cancer cell with an IDH-1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1.

The method of embodiment 19, wherein the patient is diagnosed with a cancer harboring an IDH-1 R132 mutation in a cell obtained from the patient, prior to the administration of Compound 1.

A method of treating a patient diagnosed with a cancer harboring a cancer cell with an IDH-1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1.

The method of embodiment 21, wherein the patient is diagnosed with a cancer harboring an IDH-1 R132 mutation in a cell obtained from the patient, prior to the administration of Compound 1.

A method of treating a patient diagnosed with a cancer, the method comprising

a. diagnosing the patient as having a mutant IDH-1 mutation in a cell obtained from the patient; and

b. administering a therapeutically effective amount of a pharmaceutical composition comprising Compound 1 to the patient in need of an inhibitor of the mutant IDH-1 enzyme that targets the mutant IDH-1 variants R132C at no greater than about 5 times the level of R132H; and

c. continuing to administer the pharmaceutical composition to the patient throughout a course of treatment of at least 6 months.

The method of embodiment 23, wherein the patient is in need of an inhibitor of mIDH-1 variants selected from the group consisting of R132L, R132G, and R132S;

The method of any one of embodiments 23-24, wherein the relative targeting of R132C and R132H variants of mIDH-1 is measured by the ratio of ICso values obtained using the assay of Example 3.

The method of any one of embodiments 23-25, wherein the patient is diagnosed as having an IDH-1 mutation in a cell from the patient using a next-generation sequencing (NGS)-based tumor genotyping assay.

The method of any one of embodiments 23-26, wherein the pharmaceutical composition is administered to the patient twice per day. The method of any one of embodiments 23-27, wherein the pharmaceutical composition is administered to the patient in a dose of 150 mg BID on consecutive days throughout the course of treatment.

The method of any one of embodiments 23-28, wherein Compound 1 in the

pharmaceutical composition has the solid form obtained from Example 5.

A method of inhibiting the production of inhibiting the production of 2-HG in a R132C mutated IDH-1 enzyme at no more than about 5 times the inhibition of 2-HG production in a R132H mutated IDH-1 enzyme, the method comprising contacting an IDH-1 enzyme not having arginine at position 132 with a composition comprising Compound 1 under conditions and for a time effective to inhibit 2-HG production in either an IDH-1 R132C or an IDH-1 R132H mutation in the mIDH-1 enzyme.

A method of treating a cancer in an adult patient, the cancer having a known mIDH-1 frequency of about 10-90%, the method comprising administering to a patient diagnosed with an IDH-1 mutation comprising an IDH-1 mutation selected from the group consisting of R132C, R132H, R132L, R132G, and R132S, the method comprising administering to the patient in need thereof a pharmaceutical composition comprising a total of 150 mg of a pharmaceutically acceptable solid form of 5-{[(lS)-l-(6-chloro-2- oxo- 1 ,2-dihydroquinolin-3 -yl)ethyl]amino} - 1 -methyl-6-oxo- 1 ,6-dihydropyridine-2- carbonitrile, twice per day on consecutive days for a course of treatment comprising 6 months.

The method of embodiment 31, wherein the patient is diagnosed as having an IDH-1 mutation in a cell from the patient using a next-generation sequencing (NGS)-based tumor genotyping assay.

The method of any one of embodiments 31-32, wherein the pharmaceutical composition is administered to the patient every 12 hours.

The method of any one of embodiments 31-33, wherein Compound 1 in the

pharmaceutical composition has the solid form obtained from Example 5.

A method of treating a chrondrosarcoma cancer having an IDH-1 mutation in an adult patient, the method comprising administering to the patient in need thereof a

pharmaceutical composition comprising a total of 150 mg of a pharmaceutically acceptable solid form of 5-{[(lS)-l-(6-chloro-2-oxo-l,2-dihydroquinolin-3- yl)ethyl]amino}-l-methyl-6-oxo-l,6-dihydropyridine-2-carbonitrile, twice per day on consecutive days for a course of treatment comprising 6 months.

The method of embodiment 35, wherein the patient is diagnosed as having an IDH-1 mutation in a cell from the patient using a next-generation sequencing (NGS)-based tumor genotyping assay.

The method of any one of embodiments 35-36, wherein the pharmaceutical composition is administered to the patient every 12 hours.

A method of treating a patient diagnosed with a form of cancer characterized by an IDH1 mutation selected from the group consisting of R132G, R132S and R132L, the method comprising orally administering a total of 150 mg of Compound 1 twice per day (e.g. only twice per day) to the patient in need thereof:

(Compound 1).

The method of embodiment 38, wherein the patient is diagnosed with a cancer characterized by a concurrent mutation selected from the group consisting of FLT3, NPM1, CEBPA and TP53.

The method of embodiment 38, wherein the patient is diagnosed with a cancer that is not characterized by an IDH2 mutation.

The method of embodiment 38, wherein the patient is diagnosed with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS) characterized by the IDH1 mutation.

The method of embodiment 41, wherein the patient is diagnosed with MDS or AML further characterized by a concurrent mutation selected from the group consisting of FLT3, NPM1, CEBPA and TP53.

The method of embodiment 38, wherein the patient is diagnosed with a cancer characterized by a concurrent mutation selected from the group consisting of DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXL1, FLT3, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53.

A method of treating a patient diagnosed with a hematological malignancy characterized by an IDH1 mutation selected from the group consisting of R132C, R123H, R132G, R132S and R132L and a concurrent FLT3 mutation, the method comprising

administering a total of 150 mg of Compound 1 orally twice per day (e.g. only twice per day) to the patient in need thereof:

(Compound 1).

The method of embodiment 44, wherein the patient is diagnosed with a hematological malignancy characterized by a co-mutation selected from the group consisting of DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXL1, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53.

The method of embodiment 44, comprising administering Compound 1 to the patient every day for 6 months.

The method of embodiment 44, wherein Compound 1 is administered to the patient as a single agent for the treatment of AML.

The method of embodiment 44, wherein Compound 1 is administered to the patient in combination with azacitidine during one or more 28-day treatment cycles, wherein

a. the azacitidine is administered to the patient at the dose of 75 mg/m² for 7 days IV/SC per every 28-day cycle; and

b. a total of 150 mg of Compound 1 is administered to the patient twice per day throughout the 28-day treatment cycles.

The method of embodiment 44, wherein the patient does not have an IDH-2 mutation. A method of treating a patient diagnosed with a form of cancer characterized by an IDH1 mutation selected from the group consisting of R132G, R132S and R132L, the method comprising orally administering to the patient in need thereof a total amount of 150 mg of Compound 1 BID to the patient in need thereof:

(Compound 1),

each day for a total of at least 6 months to treat the cancer characterized by the IDH1 mutation.

51. The method of embodiment 50, wherein the patient is diagnosed with a cancer

characterized by a concurrent FLT3 mutation.

52. The method of embodiment 50, wherein the patient is diagnosed with a cancer

characterized by a concurrent NPM1 mutation.

53. The method of embodiment 50, wherein the patient is diagnosed with a cancer

characterized by a concurrent CEBPA mutation.

54. The method of embodiment 50, wherein the patient is diagnosed with a cancer

characterized by a concurrent TP53 mutation.

55. The method of embodiment 50, wherein the patient is diagnosed with a cancer further characterized by both the mIDHI mutation and a co-mutation selected from the group consisting of DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXL1, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53.

56. The method of embodiment 51, wherein Compound 1 is administered as a single agent without azacitidine.

57. The method of embodiment 51, wherein Compound 1 is administered in combination with azacitidine to treat the mIDHI cancer.

[0205] The present disclosure contemplates, among other things, the following numbered embodiments: A method of treating a patient diagnosed with relapsed/refractory mIDHI gliomas comprising a step of administering to the patient in need thereof a total of 150 mg of olutasidenib twice per day (BID).

The method of embodiment 1, wherein the olutasidenib is administered orally.

The method of any one of embodiments 1-2, wherein the olutasidenib is administered to the patient in need thereof without administration of azacitidine.

The method of embodiment 3, wherein the olutasidenib is administered as a single agent to treat the relap sed/refractory mIDHI glioma in the patient.

The method of embodiment 1 or embodiment 2, wherein the olutasidenib is administered to the patient in need thereof in combination with administration of azacitidine to treat the relapsed/refractory mIDHI glioma in the patient.

The method of any one of embodiments 1-5, wherein the patient has previously received temozolomide to treat the mIDHI glioma in the patient, prior to administration of the olutasidenib to the patient.

The method of any one of embodiments 1-4, wherein the olutasidenib is administered to the patient daily throughout a course of treatment having a duration of at least 0.2 months.

The method of embodiment 7, wherein the duration of treatment is between 0.2 - 11.4 months.

The method of any one of embodiments 1-4, wherein the olutasidenib is administered to the patient daily throughout a course of treatment having a duration of at least 1 month. The method of embodiment 9, wherein the duration of treatment is 1-12 months.

The method of embodiment 9, wherein the duration of treatment is 1-11.4 months.

The method of embodiment 9, wherein the duration of treatment is one year.

The method of embodiment 5 or embodiment 6, wherein the olutasidenib is administered to the patient daily throughout a course of treatment having a duration of at least 0.2 months. 14. The method of embodiment 9, wherein the duration of treatment is 0.2-2.3 months.

15. The method of embodiment 9, wherein the duration of treatment is 1-3 months.

16. A method of treating a patient diagnosed with relapsed/refractory mlDHl solid tumors comprising the step of administering to the patient in need thereof a total of 150 mg of olutasidenib twice per day (BID).

17. The method of embodiment 16, wherein the olutasidenib is administered orally.

18. The method of embodiment 16 or embodiment 17, wherein the patient is diagnosed with a mlDHl solid tumor is selected from the group consisting of intrahepatic

cholangiocarcinoma (IHCC) and chondrosarcoma (CS).

19. The method of embodiment 18, wherein the patient is diagnosed with a mlDHl

intrahepatic cholangiocarcinoma (IHCC) solid tumor.

20. The method of embodiment 18, wherein the patient is diagnosed with a mlDHl

chondrosarcoma (CS) solid tumor.

21. The method of any one of embodiments 1-20, wherein the patient has a mlDHl tumor having a mutation selected from the group consisting of: R132C, R132G, R132S, R132H, and R132L.

[0206] The present disclosure contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient diagnosed with a cancer characterized by (i) an IDH-1 mutation selection from R132G, R132S, and R132L and (ii) a mutation selected from DNMT3A,

NPM1, SRSF2, NRAS, RUNX1, ASXL1, FLT3, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53, the method comprising orally administering 150 mg of Compound 1 :

twice daily to the patient in need thereof. 2. The method of embodiment 1, wherein the cancer is not characterized by an IDH2 mutation.

3. The method of embodiment 1 or 2, wherein the cancer is characterized by a mutation selected from FLT3, NPM1, CEBPA and TP53.

4. The method of any one of embodiments 1-3, wherein the cancer is characterized by a mutation selected from DNMT3A, TP53, ATM, and NRAS.

5. The method of any one of embodiments 1-4, wherein the cancer is acute myeloid leukemia or myelodysplastic syndrome.

6. The method of any one of embodiments 1-4, wherein the cancer is glioma.

7. The method of any one of embodiments 1-6, wherein Compound 1 is administered to the patient every day for at least 6 months.

8. The method of any one of embodiments 1-7, wherein Compound 1 is administered as a single agent.

9. The method of any one of embodiments 1-7, wherein Compound 1 is administered to the patient in combination with azacitidine during one or more 28-day treatment cycles, wherein: a. the azacitidine is administered to the patient at the dose of 75 mg/m² for 7 days IV/SC per every 28-day cycle; and

b. a total of 150 mg of Compound 1 is administered BID to the patient every day throughout the one or more 28-day treatment cycles.

10. A method of treating a patient diagnosed with a hematological malignancy characterized by (i) at least one IDHl mutation selected from R132C, R132H, R132G, R132S, and R132L and (ii) at least one mutation selected from CEBPA, DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXL1, FLT3, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53, the method comprising orally administering 150 mg of Compound 1 :

as a single agent twice daily to the patient in need thereof. 11. The method of embodiment 10, wherein the hematological malignancy is with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS).

12. The method of embodiment 10 or 11, wherein the hematological malignancy is characterized by at least one IDH1 mutation selected from R132G, R132S, and R132L.

13. The method of any one of embodiments 10-12, wherein the hematological malignancy is not characterized by an IDH2 mutation.

14. The method of any one of embodiments 10-13, wherein the hematological malignancy is characterized by at least one mutation selected from FLT3, NPM1, CEBPA and TP53.

12. A method of treating a patient diagnosed with glioma characterized by (i) at least one IDH1 mutation selected from R132C, R132H, R132G, R132S, and R132L and (ii) at least one mutation selected from DNMT3 A, TP53, ATM, and NRAS, the method comprising orally administering 150 mg of Compound 1 :

twice daily to the patient in need thereof.

13. The method of embodiment 12, wherein the glioma is characterized by at least one IDH1 mutation selected from R132G, R132S, and R132L.

14. The method of embodiment 12 or 13, wherein the glioma is not characterized by an IDH2 mutation.

15. The method of any one of embodiments 12-14, wherein Compound 1 is administered as a single agent for treatment of glioma.

16. The method of any one of embodiments 12-15, wherein Compound 1 is administered to the patient every day for at least 6 months.

17. A method of treating a patient diagnosed with a hematological malignancy characterized by (i) at least one IDHl mutation selected from R132C, R132H, R132G, R132S, and R132L and (ii) at least one mutation selected from CEBPA, DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXL1, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53, the method comprising orally administering 150 mg of Compound 1 :

twice daily in combination with azacitidine to the patient in need thereof.

18. The method of embodiment 17, wherein the hematological malignancy is characterized by at least one mutation selected from NPM1, SRSF2, RUNX 1 , ASXL1, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, EED, WT1, CBL, CSF3R,

ETNK1, PTPN11, ATM and TP53.

19. The method of embodiment 17 or 18, wherein the hematological malignancy is with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS).

20. The method of any one of embodiments 17-19, wherein the hematological malignancy is characterized by at least one IDH1 mutation selected from R132G, R132S, and R132L.

21. The method of any one of embodiments 17-20, wherein the hematological malignancy is not characterized by an IDH2 mutation.

22. The method of any one of embodiments 17-21, wherein Compound 1 is administered to the patient in combination with azacitidine during one or more 28-day treatment cycles, wherein: a. the azacitidine is administered to the patient at the dose of 75 mg/m² for 7 days IV/SC per every 28-day cycle; and

b. a total of 150 mg of Compound 1 is administered BID to the patient every day throughout the one or more 28-day treatment cycles.

[0207] The present disclosure contemplates, among other things, the following numbered embodiments:

1. A method of treating AML or MDS in a patient harboring isocitrate dehydrogenase 1 mutations (mIDH-1 ), the method comprising administering to a patient in need thereof a therapeutically effective amount of a R132X mIDH-1 Selective Inhibitor Therapy consisting of Compound 1 in an oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 15 days, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having a durable therapeutically effective trough blood plasma concentration of Compound 1 in the patient throughout the course of treatment.

2. A method of treating AML or MDS in a patient harboring isocitrate dehydrogenase 1 mutations ( mIDH-1 ), the method comprising administering to a patient in need thereof a therapeutically effective amount of Compound 1 for at least three consecutive treatment cycles of 28 consecutive days each, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having both:

a. a durable sustained therapeutically effective trough blood plasma concentration of

Compound 1 in the patient throughout the course of treatment, and

b. a reduced level of 2-HG in the patient’ s plasma after the first two consecutive treatment cycles.

3. The method of any one of embodiments 1 or 2, wherein

a. the steady state blood plasma concentration of Compound 1 in the patient is maintained at or above about 2,000 ng/mL throughout the course of treatment, and

b. the level of 2-HG in the patient plasma is maintained at or below about 200 ng/mL after the start of the third consecutive treatment cycle of the course of treatment on day 15 after the initial dose of Compound 1.

4. The method of any one of embodiments 1-3, wherein the Compound 1 is administered in a dose of 150 mg/day twice per day throughout the course of treatment.

5. The method of any one of embodiments 1-4, wherein the Compound 1 is administered with food to improve bioavailability of Compound 1.

6. The method of any one of embodiments 1-5, wherein the course of treatment is 12-30 weeks.

7. The method of any one of embodiments 1-6, wherein the method further comprises the administration of azacitidine to the patient throughout the course of treatment.

8. The method of any one of embodiments 1-6, wherein the patient is receiving or

previously received treatment with azacitidine.

9. The method of embodiment 7 or 8, wherein the azacitidine is subcutaneously or

intravenously administered to the patient in an azacitidine treatment cycle consisting of the administration of a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient.

The method of any one of embodiments 1-6, wherein the method further comprises the administration of cytarabine to the patient throughout the course of treatment.

The method of any one of embodiments 1-6, wherein the patient is receiving or previously received treatment with cytarabine.

The method of embodiment 10 or 11, wherein the cytarabine is subcutaneously or intravenously administered to the patient in a cytarabine treatment cycle consisting of the administration of a total dose of 20 mg/day each day for 7 consecutive days beginning at the start of each treatment cycle, followed 10 consecutive days without administration of the cytarabine to the patient.

The method of any one of embodiment 1-12, wherein the patient has been identified as having a R132X mutation in mIDH-1 using a diagnostic method comprising a next generation sequencing (NGS) analysis of a bone marrow or other tissue sample from the patient obtained prior to the administration of Compound 1 to the patient.

The method of any one of embodiments 1-13, wherein the subject has relapsed or refractory (R/R) AML.

The method of any one of embodiments 1-13, wherein the subject has AML or MDS with residual R132X mIDH-1 in morphologic complete remission or complete remission with incomplete blood count recovery (CR/CRi) after cytotoxic-containing induction therapy with residual IDH-R132X mutation.

The method of any one of embodiments 1-13, wherein the subject has relapsed or refractory AML or MDS previously treated with an IDHl inhibitor.

The method of any one of embodiments 1-13, wherein the subject has relapsed or refractory AML or MDS that are naive to prior hypomethylating therapy and IDHl inhibitor therapy

The method of any one of embodiments 1-13, wherein the subject has relapsed or refractory AML or MDS that has inadequately responded or has progressed immediately preceding hypomethylating therapy. The method of any one of embodiments 1-13, wherein the subject is a subject with relapsed or refractory AML or MDS that have been previously treated with single-agent IDH1 inhibitor therapy as their last therapy prior to study enrollment.

The method of any one of embodiments 1-19, wherein the subject has been diagnosed with AML with a R132X mIDH-1.

The method of any one of embodiments 1-19, wherein the subject has MDS with AML with a R132X mIDH-1.

The method of any one of embodiments 1-21, wherein the subject has been diagnosed with a R132X mIDH-1 selected from the group consisting of R132H, R132C or both R132H and R132C.

A method of treating AML or MDS in a patient harboring at least one R132X isocitrate dehydrogenase 1 mutation (mIDH-1), the method comprising administering to the patient in need thereof a R132X mIDH-1 Selective Inhibitor Therapy consisting of 150 mg BID of Compound 1 in an pharmaceutically acceptable oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 15 days.

The method of embodiment 23, wherein the course of treatment comprises at least 4 consecutive treatment cycles each consisting of 28 consecutive days of administering Compound 1 to the patient twice per day.

The method of embodiment 23, wherein the course of treatment comprises 12-30 weeks of consecutive days of administering Compound 1 to the patient twice per day.

A method of treating AML or MDS in a patient harboring at least one R132X isocitrate dehydrogenase 1 mutation (mIDH-1), the method comprising administering to the patient 150 mg BID of Compound 1 in a pharmaceutically acceptable oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 15 days.

A method of treating AML or MDS in a patient harboring at least one R132X isocitrate dehydrogenase 1 mutation (mIDH-1), the method comprising administering to the patient 150 mg BID of Compound 1 in a pharmaceutically acceptable oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 12 weeks. 28. A method of treating AML or MDS in a patient harboring at least one R132X isocitrate dehydrogenase 1 mutation (mIDH-1), the method comprising administering to the patient 150 mg BID of Compound 1 in a pharmaceutically acceptable oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 30 weeks.

29. A method of treating AML or MDS in a patient harboring one or more R132X isocitrate dehydrogenase 1 mutations ( mIDH-1 ), the method comprising administering to a patient in need thereof a therapeutically effective amount of a R132X mIDH-1 Selective Inhibitor Therapy consisting of Compound 1 in an oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 15 days, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having both:

a. a durable trough blood plasma concentration of Compound 1 in the patient measured at or above the IC90 concentration of Compound 1 for 2-HG suppression of the R132X mIDH-1 throughout the course of treatment, and

b. a level of 2-HG in the patient’s plasma of less than about 200 ng/mL within two initial consecutive treatment cycles that is maintained throughout the course of treatment.

30. The method of embodiment 29, wherein course of treatment is at least 3 consecutive 28- day treatment cycles.

31. The method of embodiments 29 or 30, wherein the trough blood plasma concentration of Compound 1 in the patient is measured between 2,000 ng/mL-7,200 ng/mL.

32. The method of any one of embodiments 29-31, wherein the level of 2-HG is maintained at about 180 ng/mL in the blood plasma of the patient throughout the course of treatment after the first 15 consecutive days of administering Compound 1 to the patient.

33. The method of any one of embodiments 1-32, wherein the concentration of Compound 1 measured in the blood of the patient does not decline more than 20% throughout the course of treatment compared to the steady state blood concentration of Compound 1 measured prior to administration of Compound 1 at cycle 2, day 1 (day 29).

34. The method of any one of embodiments 1-32, wherein the concentration of Compound 1 measured in the blood of the patient does not decline more than 10% throughout the course of treatment compared to the steady state blood concentration of Compound 1 measured prior to administration of Compound 1 at cycle 2, day 1 (day 29).

The method of any one of embodiments 33 and 34, wherein the course of treatment is a total of 12 weeks.

The method of any one of embodiments 33 and 34, wherein the course of treatment is a total of 30 weeks.

The method of any one of embodiments 22-36, wherein the subject has been diagnosed with a R132X mIDH-1 selected from the group consisting of R132H, R132C or both R132H and R132C.

The method of any one of embodiments 1-37, wherein Compound 1 is administered in the pharmaceutically acceptable dosage form obtainable from Step 6 of Example 5. A method of treating AML in a patient harboring at least one R132X isocitrate dehydrogenase 1 mutation (mIDH-1) selected from the group consisting of: R132C, R132H, R132S, R132G, and R132L, wherein the method comprises administering to the patient 150 mg BID of Compound 1 in a pharmaceutically acceptable oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing for at least 28 consecutive days.

A method of treating AML in a patient harboring one or more R132X isocitrate dehydrogenase 1 mutations (mIDH-1 ) selected from the group consisting of: R132C, R132H, R132S, R132G, and R132L, wherein the method comprises administering to a patient in need thereof a therapeutically effective amount of a R132X mIDH-1 Selective Inhibitor Therapy consisting of Compound 1 in an oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing for at least 15 consecutive days, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having a durable trough blood plasma concentration of Compound 1 in the patient measured at or above the IC90 concentration of Compound 1 for 2-HG suppression of the R132X mIDH-1 throughout the course of treatment.

The method of embodiment 40, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having a level of 2-HG in the patient’s plasma of less than about 200 ng/mL within two initial consecutive treatment cycles that is maintained throughout the course of treatment.

A method of treating AML in a patient harboring one or more R132X isocitrate dehydrogenase 1 mutations (mIDH-1 ) selected from the group consisting of: R132C, R132H, R132S, R132G, and R132L, wherein the method comprises administering to a patient in need thereof a therapeutically effective amount of a R132X mIDH-1 Selective Inhibitor Therapy consisting of Compound 1 in an oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing for at least 15 consecutive days, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having a level of 2-HG in the patient’s plasma of less than about 200 ng/mL within two initial consecutive treatment cycles that is maintained throughout the course of treatment.

The method of embodiment 42, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having a durable trough blood plasma concentration of Compound 1 in the patient measured at or above the IC90 concentration of Compound 1 for 2-HG suppression of the R132X mIDH-1 throughout the course of treatment.

A method of treating AML in a patient harboring one or more R132X isocitrate dehydrogenase 1 mutations (mIDH-1 ) selected from the group consisting of: R132C, R132H, R132S, R132G, and R132L, the method comprising administering to a patient in need thereof a therapeutically effective amount of a R132X mIDH-1 Selective Inhibitor Therapy consisting of Compound 1 in an oral dosage form for a course of treatment starting with the initial administration of Compound 1 and continuing for at least 15 consecutive days, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having a durable trough blood plasma concentration of Compound 1 in the patient measured at or above 2,000 ng/mL throughout the course of treatment of at least 6 months.

The method of embodiment 44, wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having a level of 2-HG in the patient’s plasma of less than about 200 ng/mL within two initial consecutive 28-day treatment cycles and is maintained throughout the course of treatment. 44. A method of treating AML in a patient harboring one or more R132X isocitrate dehydrogenase 1 mutations (mIDH-1) selected from the group consisting of: R132C, R132H, R132S, R132G, and R132L, wherein the method comprises administering to a patient in need thereof:

a. a therapeutically effective amount of Compound 1 in an oral dosage form for a Course of Treatment starting with the initial administration of Compound 1 and continuing for at least 15 consecutive days for one or more consecutive R132X mIDH-1 Selective

Inhibitor 28-day Treatment Cycles, wherein Compound 1 is the only R132X mIDH-1 selective inhibitor administered to the patient during the Course of Treatment; and b. azacitidine subcutaneously or intravenously administered to the patient throughout the one or more R132X mIDH-1 Selective Inhibitor 28-day Treatment Cycle(s), in an azacitidine treatment cycle consisting of the administration of a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient;

wherein the administration of the therapeutically effective amount of Compound 1 results in the patient having a durable trough blood plasma concentration of Compound 1 in the patient that is no less than about 90% of the concentration of Compound 1 measured after the first 15 days of the first R132X mIDH-1 Selective Inhibitor 28-day Treatment Cycle, throughout the Course of Treatment of at least 6 months.

45. A method of treating an adult patient diagnosed with a mutant IDH-1 form of AML, comprising administering to the patient in need thereof a combination therapy throughout a Course of Treatment lasting at least 6 months, wherein the combination therapy comprises a combination of:

a. 150 mg BID of Compound 1 each day throughout a Course of Treatment of at least 6 months, wherein the Compound 1 is the only mutant IDH-1 inhibitor administered to the patient during the Course of Treatment;

b. azacitidine subcutaneously or intravenously administered to the patient throughout the Course of Treatment at a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient. 46. A method of treating an adult patient diagnosed with a mutant IDH-1 form of AML, comprising administering to the patient in need thereof 150 mg BID of Compound 1 each day throughout a Course of Treatment of at least 6 months, in combination with azacitidine for the treatment of the AML in the adult patient, wherein Compound 1 is the only mutant IDH-1 inhibitor targeting R132C, R132H, R132S, R132G, or R132L variants of IDH-1 that is administered to the patient during the Course of Treatment.

47. The method of embodiment 46, wherein the azacitidine is administered subcutaneously or intravenously to the patient throughout the Course of Treatment at a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient.

[0208] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient harboring one or more R132X isocitrate

dehydrogenase 1 mutations ( mIDH-1 ), the method comprising administering to the patient in need thereof a R132X mIDH-1 Selective Inhibitor Therapy consisting of the oral administration of Compound 1 to the patient at a dose of 150 mg twice per day (BID) for a Course of Treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 15 days.

2. The method of embodiment 1, wherein Compound 1 is administered with food to improve bioavailability of Compound 1.

3. The method of any one of embodiments 1-2, wherein the Course of Treatment is at least 12 weeks.

4. The method of any one of claims 1-2, wherein the Course of Treatment is at least 20 weeks.

5. The method of any one of embodiments 1-2, wherein the Course of Treatment is at least 30 weeks.

6. The method of any one of embodiments 1-2, wherein the Course of Treatment is at least 6 months.

7. The method of any one of embodiments 1-6, wherein the subject has a R132X mlDH- 1 selected from the group consisting of R132H, R132C or both R132H and R132C. The method of any one of embodiments 1-7, wherein Compound 1 is administered in the solid form obtained from Example 5.

A method of treatment of a disease in a patient harboring a mIDH-1 mutation, the method comprising the steps of:

a. selecting a patient for treatment based on the presence of an IDH-1 mutation

detected in cells obtained from the patient;

b. administering Compound 1 to the selected patient from step (a) at a starting dose of 150 mg taken orally twice daily until disease progression or unacceptable toxicity.

The method of embodiment 9, further comprising administering Compound 1 to the selected patient for at least 6 months.

The method of any one of embodiments 9-10, wherein Compound 1 is administered to the patient as a single agent or in combination with other therapeutic agents.

The method of any one of embodiments 9-11, further comprising assessing blood counts and blood chemistries of the selected patient for leukocytosis and tumor lysis syndrome prior to the initiation of Compound 1 in step (b).

The method of any one of embodiments 9-12, further comprising the step (c) of monitoring a selected patient at a minimum of every 2 weeks for at least the first 3 months during treatment of the patient with compound 1 according to step (b).

The method of any one of embodiments 9-13, wherein Compound 1 is administered in the solid form obtained from Example 5 in a pharmaceutically acceptable oral dosage form.

The method of any one of embodiments 9-14 wherein the patient is selected for treatment based on the detection of one or more R132X IDH-1 mutations in cells obtained from the patient.

The method of embodiment 15, wherein the R132X IDH-1 mutation is detected using next generation sequencing of patient bone marrow tissue.

The method of any one of embodiments 15-16, wherein the R132X mutation is selected from the group consisting of R132H and R132C.

The method of any one of embodiments 16-17, wherein the R132X IDH-1 mutation is detected prior to the administration of Compound 1. A method of treating a patient harboring one or more R132X isocitrate

dehydrogenase 1 mutations ( mIDH-1 ), the method comprising orally administering to the patient 150 mg of an oral pharmaceutical dosage form of the compound 5-{[(lS)- 1 -(6-chloro-2-oxo- 1 ,2-dihydroquinolin-3 -yl)ethyl]amino} - 1 -methyl-6-oxo- 1 ,6- dihydropyridine-2-carbonitrile twice per day (BID) on consecutive days for at least 15 days.

The method of embodiment 19, wherein the oral pharmaceutical dosage form is the solid form of Compound 1 obtained by the process of Example 5.

The method of any one of embodiments 19-20, wherein the R132X mutation is selected from the group consisting of R132H and R132C.

The method of any one of embodiments 19-21, wherein the compound is

administered to the patient on consecutive days for at least 20 weeks.

The method of any one of embodiments 19-22, wherein the compound is

administered to the patient on consecutive days for at least 30 weeks.

The method of any one of embodiments 19-23, further comprising detection of the R132X IDH-1 mutation in the patient using next generation sequencing of patient bone marrow tissue prior to the administration of the compound to the patient.

A method of treatment comprising orally administering to a patient harboring a R132X mIDH-1 arginine mutation a dose of 150 mg BID of a pharmaceutically acceptable solid form of compound of formula (I) as obtained from Example 5

1

The method of embodiment 25, wherein the pharmaceutically acceptable solid form of Compound 1 is administered to the patient on consecutive days for at least 20 weeks.

The method of embodiment 25, wherein the pharmaceutically acceptable solid form of Compound 1 is administered to the patient on consecutive days for at least 30 weeks. The method of embodiment 25, wherein the pharmaceutically acceptable solid form of Compound 1 is administered to the patient on consecutive days for at least 6 months.

The method of any one of embodiments 25-28, wherein Compound 1 is administered to the patient as a single agent or in combination with other therapeutic agents.

The method of any one of embodiments 25-29, wherein the R132X mIDH-1 arginine mutation is selected from the group consisting of: R132H, R132C, R132G, R132L, and R132S.

The method of any one of embodiments 25-30, further comprising detection of the R132X IDH-1 arginine mutation in the patient using next generation sequencing of patient bone marrow tissue prior to the administration of Compound 1 to the patient. The method of any one of embodiments 1-31, wherein the patient has not received another mIDH-1 inhibitor compound.

The method of any one of embodiments 1-8, wherein the patient has greater than 200 ng/mL of 2-HG prior to initiating the R132X mIDH-1 Selective Inhibitor Therapy. The method of any one of embodiments 9-18, wherein the patient selected in step (a) has greater than 200 ng/mL of 2-HG prior to administering Compound 1 in step (b). The method of any one of embodiments 19-24, wherein the patient has greater than 200 ng/mL of 2-HG prior to administration of the oral pharmaceutical dosage form of the compound to the patient.

The method of any one of embodiments 25-31, wherein the patient has greater than 200 ng/mL of 2-HG prior to administration of the compound of formula (I) to the patient.

A method of treatment comprising orally administering to a patient having elevated blood levels of 2-HG greater than about 200 ng/mL and harboring at least one of a R132H, R132C, R132G, R132L, and R132S IDH-1 mutation, a dose of 150 mg BID each day of a pharmaceutically acceptable solid form of compound of formula (I) as obtained from Example 5 1

on consecutive days for at least 15 days.

38. The method of embodiment 37, wherein the pharmaceutically acceptable solid form of Compound 1 is administered to the patient on consecutive days for at least 20 weeks.

39. The method of embodiment 37, wherein the pharmaceutically acceptable solid form of Compound 1 is administered to the patient on consecutive days for at least 30 weeks.

40. The method of embodiment 37, wherein the pharmaceutically acceptable solid form of Compound 1 is administered to the patient on consecutive days for at least 6 months.

41. A method of treating a patient diagnosed with a cancer harboring one or more R132X isocitrate dehydrogenase 1 mutations ( mIDH-1 ) selected from the group consisting of R132C, R132H, R132L, R132G, and R132S, the method comprising the oral administration of Compound 1 as the only inhibitor of mIDH-1 to the patient, at a total dose of 150 mg twice per day (BID) throughout a Course of Treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 6 months.

42. A method of treating an adult patient diagnosed with a cancer harboring one or more R132X isocitrate dehydrogenase 1 mutations ( mIDH-1 ) selected from the group consisting of R132C, R132H, R132L, R132G, and R132S, the method comprising the oral administration of a total dose of 150 mg twice per day (BID) of Compound 1 in the solid form obtained from Example 5, on consecutive days throughout a Course of Treatment starting with the initial administration of Compound 1 and continuing on consecutive days for at least 6 months, wherein Compound 1 is the only inhibitor of mIDH-1 administered to the patient during a Course of Treatment. 43. The method of any one of embodiments 41-42, further comprising the administration of an hypomethylating agent to the patient during the Course of Treatment.

[0209] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient diagnosed with a glioma cancer harboring a cancer cell with an IDH-1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1 and a hypomethylating agent over a course of treatment comprising multiple 28-day treatment cycles.

2. A method of treatment comprising the steps of:

a. selecting a patient diagnosed with a glioma cancer harboring an IDH-1 mutation;

b. administering Compound 1 to the selected patient from step (a) at a starting dose of 150 mg taken orally twice daily for a treatment cycle of 28 consecutive days; and

c. administering azacitidine subcutaneously or intravenously to the patient during the same treatment cycle as step (b), in a total dose of 75 mg/m2 each day for the first 7 consecutive days of the treatment cycle followed by 21 consecutive days without the administration of azacitidine until the end of the treatment cycle, with the exception of optionally permitting a 48-hour dose interruption of azacitidine on a weekend of holiday during the treatment cycle.

3. The method of any one of embodiments 1-2, wherein the cancer does not harbor a IDH-2 mutation.

4. The method of any one of embodiments 1-3, wherein the patient has been diagnosed as having a R132 mutation based on a patient diagnostic.

5. The method of embodiment 4, wherein the patient diagnostic comprises detecting the R132 mutation in a biological sample obtained from the patient.

6. The method of embodiment 5, wherein the tissue sample is obtained from the CNS of the patient.

7. The method of any one of embodiments 4-6, wherein the R132 mutation is detected using next generation sequencing (NGS) without the use of PCR.

8. The method of any one of embodiments 1-7, wherein the method comprises

administering 150 mg of Compound 1 to the patient in the solid form obtained from the method of Example 5. 9. The method of any one of embodiments 1-8, wherein Compound 1 is administered to the patient once every 12 hours on consecutive days throughout a course of treatment.

10. The method of any one of embodiments 1-9, wherein Compound 1 is administered to the patient throughout a course of treatment of at least 6 months.

11. The method of any one of embodiments 2-9, wherein Compound 1 is administered to the patient throughout a course of treatment of at least 6 consecutive treatment cycles.

12. A method of treating a glioma cancer having an IDH-1 mutation in an adult patient, the method comprising administering to the patient in need thereof

a. a pharmaceutical composition comprising a total of 150 mg of a pharmaceutically acceptable solid form of 5-{[(lS)-l-(6-chloro-2-oxo-l,2-dihydroquinolin-3-yl)ethyl]amino}-l-methyl-6- oxo-l,6-dihydropyridine-2-carbonitrile obtained from Example 5, twice per day on consecutive days for a 28 day treatment cycle; and

b. administering azacitidine subcutaneously or intravenously to the patient during the same treatment cycle as step (a), in a total dose of 75 mg/m² each day for the first 7 consecutive days of the treatment cycle followed by 21 consecutive days without the administration of azacitidine until the end of the treatment cycle, with the exception of optionally permitting a 48-hour dose interruption of azacitidine on a weekend of holiday during the treatment cycle.

13. A method of treating a glioma cancer having an IDH-1 mutation in an adult patient, the method comprising administering to the patient in need thereof

a. a pharmaceutical composition comprising a total of 150 mg of a Compound 1

in a pharmaceutically acceptable solid form (e.g. that obtainable from Example 5), twice per day on consecutive days for a 28 day treatment cycle; and

b. administering azacitidine subcutaneously or intravenously to the patient during the same treatment cycle as step (a), in a total dose of 75 mg/m² each day for the first 7 consecutive days of the treatment cycle followed by 21 consecutive days without the administration of azacitidine until the end of the treatment cycle, with the exception of optionally permitting a

48-hour dose interruption of azacitidine on a weekend of holiday during the treatment cycle.

14. The method of any one of embodiments 1-13, wherein the patient is diagnosed with

glioblastoma multiforme prior to the administration of Compound 1.

15. The method of any one of embodiments 1-14, wherein the patient has elevated 2HG

blood levels prior to the administration of Compound 1.

16. The method of embodiment 15, wherein the level of 2HG measured in the blood of the patient is greater than about 200 ng/mL prior to the administration of Compound 1.

17. The method of any one of embodiments 1-16, wherein the level of 2HG measured in the blood of the patient is less than about 100 ng/mL measured on day 15 after the administration of Compound 1 for the first 14 consecutive days of the first treatment cycle.

[0210] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient diagnosed with a non-CNS solid tumor harboring a cancer cell with an IDH1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1 as a single agent.

2. A method of treating a patient diagnosed with glioma harboring a cancer cell with an IDH1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount Compound 1 in combination with a therapeutically effective amount of azacitidine.

3. A method of treating a patient diagnosed with a chondrosarcoma cancer harboring a cancer cell with an IDHl R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1 in combination with a therapeutically effective amount of azacitidine.

4. A method of treating a patient diagnosed with a hepatobiliary cancer harboring a cancer cell with an IDHl R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1 in combination with a therapeutically effective amount of a PD-1 inhibitor.

5. A method of treating a patient diagnosed with an intrahepatic cholangiocarcinoma cancer harboring a cancer cell with an IDHl R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of

Compound 1 in combination with a therapeutically effective amount of a gemcitabine and cisplatin chemotherapy.

The method of any one of embodiments 1-5, wherein Compound 1 is administered at a dose of 150 mg taken orally twice daily.

The method of any one of embodiments 1-6, wherein Compound 1 is orally administered as the solid form obtained from Example 5.

The method of any one of embodiments 1-7, further comprising the steps of:

a. selecting a patient diagnosed with the cancer harboring an IDHl mutation;

b. administering Compound 1 to the selected patient from step (a) at a starting dose of 150 mg taken orally twice daily for a treatment cycle of 28 consecutive days. A method of treating a solid tumor or CNS cancer having an IDHl mutation in an adult patient, the method comprising administering to the patient in need thereof a

pharmaceutical composition comprising a total of 150 mg of a pharmaceutically acceptable solid form of 5-{[(lS)-l-(6-chloro-2-oxo-l,2-dihydroquinolin-3- yl)ethyl]amino}-l-methyl-6-oxo-l,6-dihydropyridine-2-carbonitrile obtained from Example 5, twice per day on consecutive days for a 28 day treatment cycle.

The method of any one of embodiments 1-9, wherein the cancer does not harbor a IDH2 mutation.

The method of any one of embodiments 1-10, wherein the cancer does not harbor a IDH2 mutation selected from the group consisting of: IDH2 R172K and IDH2 R140Q.

The method of any one of embodiments 1-11, comprising the step of detecting the IDHl mutation in a cell from the patient using a next-generation sequencing (NGS)-based tumor genotyping assay.

The method of any one of embodiments 1-12, wherein administration of Compound 1 to the patient results in a decreased 2-hydroxyglutarate (2-HG) levels in the blood of the patient after the first 15 consecutive days of treatment of the patient with Compound 1. The method of any one of embodiments 1-13, wherein the method comprises

administering 150 mg of Compound 1 to the patient in the solid form obtained from the method of Example 5. The method of any one of embodiments 1-14, wherein the method comprises

administering 150 mg of Compound 1 to the patient twice daily throughout a course of treatment.

The method of embodiment 15, wherein the course of treatment is at least 15 consecutive days.

The method of any one of embodiments 1-16, wherein the Compound 1 is administered to the patient once every 12 hours on consecutive days throughout a course of treatment. The method of any one of embodiments 1-17, wherein the Compound 1 is administered to the patient throughout a course of treatment of at least 4 months.

The method of any one of embodiments 1-17, wherein the Compound 1 is administered to the patient throughout a course of treatment of at least 6 months.

A method of treating a patient diagnosed with a cancer selected from the group consisting of: glioma, chondrosarcoma, hepatobiliary, and intrahepatic cholangiocarcinoma, the cancer harboring an IDH1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of a mIDHI Inhibitor Therapy throughout a Course of Treatment of at least one 28-day treatment cycle, the mIDHI Inhibitor Therapy consisting of

a. Compound 1 in combination with azacitidine for the patient diagnosed with

glioma or chondrosarcoma cancer; or

b. Compound 1 in combination with a PD-1 inhibitor for the patient diagnosed with hepatobiliary cancer; or

c. Compound 1 in combination with gemcitabine and cisplatin chemotherapy for the patient diagnosed with intrahepatic cholangiocarcinoma.

The method of embodiment 20, wherein Compound 1 is administered at a dose of 150 mg BID.

The method of any one of embodiments 20-21, wherein the PD-1 inhibitor is nivolumab. The method of any one of embodiments 20-22, wherein the nivolumab is administered at a dose of 240 mg every 2 weeks or 480 mg every 4 weeks.

The method of any one of embodiments 20-23, wherein the azacitidine is administered at a dose of 75 mg/m², SC, dl-7, q4 wk throughout the Course of Treatment. 25. The method of any one of embodiments 20-24, wherein the Course of Treatment is at least 4 months.

26. The method of any one of embodiments 20-25, wherein the Course of Treatment is at least 6 months.

27. The method of any one of embodiments 1-26, wherein the Compound 1 is administered as an oral dosage form obtained from Example 5.

[0211] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient diagnosed with a chondrosarcoma cancer harboring a cancer cell with an IDHl R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1 in combination with a therapeutically effective amount of azacitidine.

2. The method of embodiment 1, wherein Compound 1 is administered at a dose of 150 mg taken orally twice daily.

3. The method of any one of embodiments 1-2, wherein Compound 1 is orally administered as the solid form obtained from Example 5.

4. The method of any one of embodiments 1-3, further comprising the steps of:

a. selecting a patient diagnosed with the cancer harboring an IDHl mutation;

b. administering Compound 1 to the selected patient from step (a) at a starting dose of 150 mg taken orally twice daily for a treatment cycle of 28 consecutive days.

5. The method of any one of embodiments 1-4, wherein the cancer does not harbor a IDH2 mutation.

6. The method of any one of embodiments 1-5, wherein the cancer does not harbor a IDH2 mutation selected from the group consisting of: IDH2 R172K and IDH2 R140Q.

7. The method of any one of embodiments 1-6, comprising the step of detecting the IDHl mutation in a cell from the patient using a next-generation sequencing (NGS)-based tumor genotyping assay.

8. The method of any one of embodiments 1-7, wherein administration of Compound 1 to the patient results in a decreased 2-hydroxyglutarate (2-HG) levels in the blood of the patient after the first 15 consecutive days of treatment of the patient with Compound 1. The method of any one of embodiments 1-8, wherein the method comprises

administering 150 mg of Compound 1 to the patient in the solid form obtained from the method of Example 5.

The method of any one of embodiments 1-9, wherein the method comprises

administering 150 mg of Compound 1 to the patient twice daily throughout a course of treatment.

The method of embodiment 10, wherein the course of treatment is at least 15 consecutive days.

The method of any one of embodiments 1-11, wherein the Compound 1 is administered to the patient once every 12 hours on consecutive days throughout a course of treatment. The method of any one of embodiments 1-12, wherein the Compound 1 is administered to the patient throughout a course of treatment of at least 4 months.

The method of any one of embodiments 1-12, wherein the Compound 1 is administered to the patient throughout a course of treatment of at least 6 months.

A method of treating a patient diagnosed with a cancer, wherein the cancer is

chondrosarcoma, the cancer harboring an IDHl R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of a mIDHI Inhibitor Therapy throughout a Course of Treatment of at least one 28-day treatment cycle, the mIDHI Inhibitor Therapy consisting of Compound 1 in combination with azacitidine for the patient diagnosed with glioma or chondrosarcoma cancer.

The method of embodiment 15, wherein Compound 1 is administered at a dose of 150 mg BID.

The method of any one of embodiments 15-16, wherein the azacitidine is administered at a dose of 75 mg/m², SC, dl-7, q4 wk throughout the Course of Treatment.

The method of any one of embodiments 15-17, wherein the Course of Treatment is at least 4 months.

The method of any one of embodiments 15-18, wherein the Course of Treatment is at least 6 months.

The method of any one of embodiments 1-19, wherein the Compound 1 is administered as an oral dosage form obtained from Example 5. [0212] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient diagnosed with an intrahepatic cholangiocarcinoma cancer harboring a cancer cell with an IDH1 R132 mutation, the method comprising

administering to the patient in need thereof a therapeutically effective amount of

Compound 1 in combination with a therapeutically effective amount of a gemcitabine and cisplatin chemotherapy.

2. The method of embodiment 1, wherein Compound 1 is administered at a dose of 150 mg taken orally twice daily.

3. The method of any one of embodiments 1-2, wherein Compound 1 is orally administered as the solid form obtained from Example 5.

4. The method of any one of embodiments 1-3, further comprising the steps of:

a. selecting a patient diagnosed with the cancer harboring an IDHl mutation;

b. administering Compound 1 to the selected patient from step (a) at a starting dose of 150 mg taken orally twice daily for a treatment cycle of 28 consecutive days.

5. The method of any one of embodiments 1-4, wherein the cancer does not harbor a IDH2 mutation.

6. The method of any one of embodiments 1-5, wherein the cancer does not harbor a IDH2 mutation selected from the group consisting of: IDH2 R172K and IDH2 R140Q.

7. The method of any one of embodiments 1-6, comprising the step of detecting the IDHl mutation in a cell from the patient using a next-generation sequencing (NGS)-based tumor genotyping assay.

8. The method of any one of embodiments 1-7, wherein administration of Compound 1 to the patient results in a decreased 2-hydroxyglutarate (2-HG) levels in the blood of the patient after the first 15 consecutive days of treatment of the patient with Compound 1.

9. The method of any one of embodiments 1-8, wherein the method comprises

administering 150 mg of Compound 1 to the patient in the solid form obtained from the method of Example 5.

10. The method of any one of embodiments 1-9, wherein the method comprises

administering 150 mg of Compound 1 to the patient twice daily throughout a course of treatment. 11. The method of embodiment 10, wherein the course of treatment is at least 15 consecutive days.

12. The method of any one of embodiments 1-11, wherein the Compound 1 is administered to the patient once every 12 hours on consecutive days throughout a course of treatment.

13. The method of any one of embodiments 1-12, wherein the Compound 1 is administered to the patient throughout a course of treatment of at least 4 months.

14. The method of any one of embodiments 1-12, wherein the Compound 1 is administered to the patient throughout a course of treatment of at least 6 months.

15. A method of treating a patient diagnosed with a cancer, wherein the cancer is intrahepatic cholangiocarcinoma, the cancer harboring an IDH1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of a mIDHI Inhibitor Therapy throughout a Course of Treatment of at least one 28-day treatment cycle, the mIDHI Inhibitor Therapy consisting of Compound 1 in combination with gemcitabine and cisplatin chemotherapy for the patient diagnosed with intrahepatic cholangiocarcinoma.

16. The method of embodiment 15, wherein Compound 1 is administered at a dose of 150 mg BID.

17. The method of any one of embodiments 15-16, wherein the Course of Treatment is at least 4 months.

18. The method of any one of embodiments 15-17, wherein the Course of Treatment is at least 6 months.

19. The method of any one of embodiments 1-18, wherein the Compound 1 is administered as an oral dosage form obtained from Example 5.

[0213] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient diagnosed with a hepatobiliary cancer harboring a cancer cell with an IDH1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of Compound 1 in combination with a therapeutically effective amount of a PD-1 inhibitor.

2. The method of embodimentl, wherein Compound 1 is administered at a dose of 150 mg taken orally twice daily. The method of any one of embodiments 1-2, wherein Compound 1 is orally administered as the solid form obtained from Example 5.

The method of any one of embodiments 1-3, further comprising the steps of:

a. selecting a patient diagnosed with the cancer harboring an IDHl mutation;

b. administering Compound 1 to the selected patient from step (a) at a starting dose of 150 mg taken orally twice daily for a treatment cycle of 28 consecutive days. The method of any one of embodiments 1-4, wherein the cancer does not harbor a IDH2 mutation.

The method of any one of embodiments 1-5, wherein the cancer does not harbor a IDH2 mutation selected from the group consisting of: IDH2 R172K and IDH2 R140Q.

The method of any one of embodiments 1-6, comprising the step of detecting the IDHl mutation in a cell from the patient using a next-generation sequencing (NGS)-based tumor genotyping assay.

The method of any one of embodiments 1-7, wherein administration of Compound 1 to the patient results in a decreased 2-hydroxyglutarate (2-HG) levels in the blood of the patient after the first 15 consecutive days of treatment of the patient with Compound 1. The method of any one of embodiments 1-8, wherein the method comprises

administering 150 mg of Compound 1 to the patient in the solid form obtained from the method of Example 5.

The method of any one of embodiments 1-9, wherein the method comprises

administering 150 mg of Compound 1 to the patient twice daily throughout a course of treatment.

The method of embodiment 10, wherein the course of treatment is at least 15 consecutive days.

The method of any one of embodiments 1-11, wherein the Compound 1 is administered to the patient once every 12 hours on consecutive days throughout a course of treatment. The method of any one of embodiments 1-12, wherein the Compound 1 is administered to the patient throughout a course of treatment of at least 4 months.

The method of any one of embodiments 1-12, wherein the Compound 1 is administered to the patient throughout a course of treatment of at least 6 months. 15. A method of treating a patient diagnosed with a cancer, wherein the cancer is hepatobiliary cancer, the cancer harboring an IDH1 R132 mutation, the method comprising administering to the patient in need thereof a therapeutically effective amount of a mIDHI Inhibitor Therapy throughout a Course of Treatment of at least one 28-day treatment cycle, the mIDHI Inhibitor Therapy consisting of Compound 1 in combination with a PD-1 inhibitor for the patient diagnosed with hepatobiliary cancer.

16. The method of embodiment 15, wherein Compound 1 is administered at a dose of 150 mg

BID.

17. The method of any one of embodiments 15-16, wherein the PD-1 inhibitor is nivolumab.

18. The method of any one of embodiments 15-17, wherein the nivolumab is administered at a dose of 240 mg every 2 weeks or 480 mg every 4 weeks.

19. The method of any one of embodiments 15-18, wherein the Course of Treatment is at least 4 months.

20. The method of any one of embodiments 15-19, wherein the Course of Treatment is at least 6 months.

21. The method of any one of embodiments 1-20, wherein the Compound 1 is administered as an oral dosage form obtained from Example 5.

[0214] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. Use of a pharmaceutical composition comprising Compound 1, or pharmaceutically

acceptable salt thereof,

1

in treating a cancer harboring an isocitrate dehydrogenase- 1 (IDH-1) mutation (mIDH-1) in a patient by administering a total of 300 mg of Compound 1 (or a corresponding amount in the form of a pharmaceutically acceptable salt thereof) to a patient each day during a course of treatment. The use of embodiment 1, wherein a 150 mg amount of Compound 1 (or a corresponding amount in the form of a pharmaceutically acceptable salt thereof) is administered to the patient twice per day (BID) throughout the course of treatment.

The use of embodiment 1, wherein the cancer is a mIDH-1 form of acute myeloid leukemia.

The use of embodiment 3, wherein the acute myeloid leukemia is relapsed or refractory or is drug-resistant.

The use of embodiment 1, wherein the cancer is a mIDH-1 solid tumor.

The use of embodiment 1, wherein the cancer is of a mIDH-1 glioma.

The use of embodiment 6, wherein the mIDH-1 glioma is an advanced glioma that has recurred or progressed prior to the administration of Compound 1.

The use of any one of the preceding embodiments, wherein the mlDHl is a R132X mutation.

The use of claim 8, wherein the R132X mIDH-1 mutation is selected from R132L, R132G and R132S.

The use of any one of the preceding embodiments, wherein the pharmaceutical composition comprising Compound 1 or a pharmaceutically acceptable salt thereof is orally administered to the patient.

The use of any one of the preceding embodiments, wherein Compound 1, (or a corresponding amount in the form of a pharmaceutically acceptable salt thereof)is administered as a single agent for the treatment of the cancer harboring the IDH-1 mutation.

The use of any of the preceding embodiments, wherein the course of treatment is at least 15 consecutive days to reach a steady state blood concentration of Compound 1 (or a corresponding amount in the form of a pharmaceutically acceptable salt thereof)in the patient.

The use of any one of the preceding embodiments, wherein the course of treatment is at least 6 months.

The use of any one of the preceding embodiments, wherein the pharmaceutical composition comprises Compound 1 in a Type A solid form characterized by a reflection X-ray powder diffraction (XRPD) pattern comprising characteristic peaks at 6.3, 12.8, 13.8, 23.6, and 27.8 degrees ±0.2° 2Q.

15. The use of any one of the preceding embodiments, wherein the pharmaceutical

composition comprises the following formulation for oral administration: (a) Type A solid form of Compound 1 in a relative weight of about 33, (b) a microcrystalline cellulose in a relative weight of about 61, (c) a croscamellose sodium in a relative weight of about 5 and a magnesium stearate in a relative weight of about 1;

wherein the Type A solid form of Compound 1 is characterized by a reflection X-ray powder diffraction (XRPD) pattern comprising characteristic peaks at 6.3, 12.8, 13.8, 23.6, and 27.8 degrees ±0.2° 2Q.

[0215] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A pharmaceutical composition comprising Compound 1 :

1

or pharmaceutically acceptable salt thereof, for use in treating a patient diagnosed with a form of cancer harboring an isocitrate dehydrogenase- 1 (IDH-1) mutation (mIDH-1) by administering a total of 300 mg of Compound 1 (or a corresponding amount in the form of a pharmaceutically acceptable salt thereof) to the patient each day during a course of treatment.

2. The pharmaceutical composition of embodiment 1, wherein a 150 mg amount of

Compound 1 (or a corresponding amount in the form of a pharmaceutically acceptable salt thereof) is administered to the patient twice per day (BID) throughout the course of treatment.

3. The pharmaceutical composition of embodiment 1, wherein the cancer is a mIDH-1 form of acute myeloid leukemia.

4. The pharmaceutical composition of embodiment 3, wherein the acute myeloid leukemia is relapsed or refractory or is drug-resistant. The pharmaceutical composition of embodiment 1, wherein the cancer is a mIDH-1 solid tumor.

The pharmaceutical composition of embodiment 1, wherein the cancer is of a mIDH-1 glioma.

The pharmaceutical composition of embodiment 6, wherein the mIDH-1 glioma is an advanced glioma that has recurred or progressed prior to the administration of Compound 1

The pharmaceutical composition of any one of the preceding embodiments, wherein the mIDHI is a R132X mutation.

The pharmaceutical composition of embodiment 8, wherein the R132X mIDH-1 mutation is selected from R132L, R132G and R132S.

The pharmaceutical composition of any one of the preceding embodiments, wherein the pharmaceutical composition comprising Compound 1 or a pharmaceutically acceptable salt thereof is orally administered to the patient.

The pharmaceutical composition of any one of the preceding embodiments, wherein Compound 1, (or a corresponding amount in the form of a pharmaceutically acceptable salt thereof) is administered as a single agent for the treatment of the cancer harboring the IDH-1 mutation.

The pharmaceutical composition of any of the preceding embodiments, wherein the course of treatment is at least 15 consecutive days to reach a steady state blood concentration of Compound 1 (or a corresponding amount in the form of a

pharmaceutically acceptable salt thereof) in the patient.

The pharmaceutical composition of any one of the preceding embodiments, wherein the course of treatment is at least 6 months.

The pharmaceutical composition of any one of the preceding embodiments, wherein the pharmaceutical composition comprises Compound 1 in a Type A solid form

characterized by a reflection X-ray powder diffraction (XRPD) pattern comprising characteristic peaks at 6.3, 12.8, 13.8, 23.6, and 27.8 degrees ±0.2° 2Q.

The pharmaceutical composition of any one of the preceding embodiments, wherein the pharmaceutical composition comprises the following formulation for oral administration: (a) Type A solid form of Compound 1 in a relative weight of about 33, (b) a microcrystalline cellulose in a relative weight of about 61, (c) a croscamellose sodium in a relative weight of about 5 and a magnesium stearate in a relative weight of about 1; wherein the Type A solid form of Compound 1 is characterized by a reflection X-ray powder diffraction (XRPD) pattern comprising characteristic peaks at 6.3, 12.8, 13.8, 23.6, and 27.8 degrees ±0.2° 2Q.

[0216] The present disclosure also contemplates, among other things, the following numbered embodiments:

1. A method of treating a patient diagnosed with a form of cancer harboring a R132X IDH1 mutation, the method comprising administering to the patient in need thereof a total of 150 mg of the compound of Formula (1) twice per day (BID)

1

2. The method of embodiment 1 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

3. The method of embodiment 1 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

4. The method of any one of the enumerated embodiments above, wherein the compound of Formula (1) is orally administered to the patient.

5. The method of any one of the enumerated embodiments above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient throughout a course of treatment of at least 15 consecutive days.

6. The method of embodiment 5 above, further comprising administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

7. The method of embodiment 5 above, further comprising administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a 6 month course of treatment. 8. The method of any one of the enumerated embodiments 1-7 above, wherein the compound of Formula (1) is administered to the patient in an oral unit dosage form.

9. A method of treating a patient diagnosed with acute myeloid leukemia (AML) having an IDH1 mutation, the method comprising administering to the patient in need thereof a combination of azacitidine and the compound of Formula (1),

1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day, and the azacitidine is administered to the patient at a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient.

10. The method of embodiment 9 above, wherein the acute myeloid leukemia is relapsed or refractory or is drug-resistant.

11. The method of any one of the embodiments 9-10 above, wherein the IDHl mutation is a R132X IDHl mutation.

12. The method of any one of the embodiments 9-11 above, wherein the R132X IDHl

mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

13. The method of any one of the embodiments 9-11 above, wherein the R132X IDHl

mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

14. The method of any one of the embodiments 9-13 above, further comprising administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

15. The method of any one of the embodiments 9-13 above, further comprising administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days. 16. The method of any one of the embodiments 9-13 above, further comprising administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

17. A method of treating a patient diagnosed with myelodysplastic syndrome (MDS) having an IDHl mutation, the method comprising administering to the patient in need thereof a combination of azacitidine and the compound of Formula (1),

1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day, and the azacitidine is administered to the patient at a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient.

18. The method of embodiment 17 above, wherein the IDHl mutation is a R132X IDHl mutation.

19. The method of embodiment 18 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

20. The method of embodiment 18 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

21. The method of any one of the embodiments 17-20 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

22. The method of any one of the embodiments 17-20 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

23. The method of any one of the embodiments 17-20 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months. 24. A method of treating a patient diagnosed with myelodysplastic syndrome (MDS) having an IDHl mutation, the method comprising administering to the patient in need thereof the compound of Formula (1),

1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day.

25. The method of embodiment 24 above, wherein the acute myeloid leukemia is relapsed or refractory or is drug-resistant.

26. The method of any one of the embodiments 24-25 above, wherein the IDHl mutation is a R132X IDHl mutation.

27. The method of embodiment 26 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

28. The method of embodiment 26 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

29. The method of any one of the embodiments 24-28 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

30. The method of any one of the embodiments 24-28 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

31. The method of any one of the embodiments 24-28 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

32. A method of treating a patient diagnosed with glioma having an IDHl mutation, wherein the method comprises administering to the patient in need thereof the compound of Formula (1), 1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day.

33. The method of any one of the embodiments 32 above, wherein the IDH1 mutation is a R132X IDH1 mutation.

34. The method of embodiment 33 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

35. The method of embodiment 34 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

36. The method of any one of the embodiments 32-35 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

37. The method of any one of the embodiments 32-35 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

38. The method of any one of the embodiments 32-35 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

39. A method of treating a patient diagnosed with chondrosarcoma having an IDHl mutation, wherein the method comprises administering to the patient in need thereof the compound of Formula (1),

1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day. 40. The method of any one of the embodiments 39 above, wherein the IDH1 mutation is a R132X IDH1 mutation.

41. The method of embodiment 40 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

42. The method of embodiment 40 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

43. The method of any one of the embodiments 39-42 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

44. The method of any one of the embodiments 39-42 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

45. The method of any one of the embodiments 39-42 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

46. A method of treating a patient diagnosed with hepatobiliary cholangiocarcinoma having an IDHl mutation, wherein the method comprises administering to the patient in need thereof the compound of Formula (1),

1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day.

47. The method of embodiment 46 above, wherein the IDHl mutation is a R132X IDHl mutation.

48. The method of embodiment 47 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

49. The method of embodiment 47 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations. 50. The method of any one of the embodiments 46-49 above, further comprising administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

51. The method of any one of the embodiments 46-49 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

52. The method of any one of the embodiments 46-49 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

53. A method of treating a patient diagnosed with intrahepatic cholangiocarcinoma having an IDH1 mutation, wherein the method comprises administering to the patient in need thereof the compound of Formula (1),

1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day.

54. The method of embodiment 53 above, wherein the IDHl mutation is a R132X IDH1 mutation.

55. The method of embodiment 54 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

56. The method of embodiment 54 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

57. The method of any one of the embodiments 53-56 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

58. The method of any one of the embodiments 53-56 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days. 59. The method of any one of the embodiments 53-56 above, further comprising administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

60. A method of treating a patient diagnosed with glioma having an IDH1 mutation, wherein the method comprises administering to the patient in need thereof a combination of azacitidine and the compound of Formula (1),

1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day, and the azacitidine is administered to the patient at a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient.

61. The method of embodiment 60 above, wherein the IDHl mutation is a R132X IDH1 mutation.

62. The method of embodiment 61 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

63. The method of embodiment 61 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

64. The method of any one of the embodiments 60-63 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

65. The method of any one of the embodiments 60-63 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

66. The method of any one of the embodiments 60-63 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months. 67. A method of treating a patient diagnosed with chondrosarcoma having an IDH1 mutation, wherein the method comprises administering to the patient in need thereof a combination of azacitidine and the compound of Formula (1),

1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day, and the azacitidine is administered to the patient at a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient.

68. The method of embodiment 67 above, wherein the IDHl mutation is a R132X IDH1 mutation.

69. The method of embodiment 68 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

70. The method of embodiment 68 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

71. The method of any one of the embodiments 67-70 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

72. The method of any one of the embodiments 67-70 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

73. The method of any one of the embodiments 67-70 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

74. A method of treating a patient diagnosed with hepatobiliary cholangiocarcinoma having an IDHl mutation, wherein the method comprises administering to the patient in need thereof a combination of azacitidine and the compound of Formula (1), 1

wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day, and the azacitidine is administered to the patient at a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient.

75. The method of embodiment 74 above, wherein the IDHl mutation is a R132X IDH1 mutation.

76. The method of embodiment 75 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

77. The method of embodiment 75 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

78. The method of any one of the embodiments 74-77 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

79. The method of any one of the embodiments 74-77 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

80. The method of any one of the embodiments 74-77 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

81. A method of treating a patient diagnosed with intrahepatic cholangiocarcinoma having an IDHl mutation, wherein the method comprises administering to the patient in need thereof a combination of azacitidine and the compound of Formula (1),

1 wherein a total of 150 mg of the compound of Formula (1) is administered to the patient twice per day (BID) each day, and the azacitidine is administered to the patient at a total dose of 75 mg/m² each day for 7 consecutive days beginning at the start of each treatment cycle, followed 21 consecutive days without administration of the azacitidine to the patient.

82. The method of embodiment 81 above, wherein the IDHl mutation is a R132X IDH1 mutation.

83. The method of embodiment 82 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132H and R132C IDHl mutations.

84. The method of embodiment 82 above, wherein the R132X IDHl mutation is selected from the group consisting of: R132S, R132G, and R132L IDHl mutations.

85. The method of any one of the embodiments 81-84 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of between 15 days and 6 months.

86. The method of any one of the embodiments 81-84 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 15 days.

87. The method of any one of the embodiments 81-84 above, further comprising

administering a total of 150 mg BID of the compound of Formula (1) to the patient on consecutive days throughout a course of treatment of at least 6 months.

88. The method of any one of the embodiments 1-87 above, wherein the compound of Formula (1) is administered in a crystalline form.

89. The method of any one of the embodiments 1-88 above, wherein the compound of Formula (1) is administered as a Type A crystalline form.

90. The method of any one of the embodiments 1-89 above, wherein the compound of Formula (1) is orally administered in a capsule comprising a total of 150 mg of the compound of Formula (1).

91. The method of any one of the embodiments 1-90 above, wherein the compound of Formula (1) is orally administered in multiple capsules each comprising a total of 50 mg of the compound of Formula (1).

[0217] The present disclosure also contemplates, among other things, the following numbered embodiments: 1. A method of treating acute myeloid leukemia (AML) in patients with an isocitrate dehydrogenase- 1 (IDH1) mutation, the method comprising steps of:

a. isolating and purifying DNA from a sample obtained from a patient;

b. detecting an IDHl mutation in the DNA from the sample; and

c. administering to the patient with the IDHl mutation a total of 150 mg of olutasidenib twice daily in a pharmaceutically acceptable composition.

2. The method of embodiment 1, wherein the IDHl mutation is an IDHl R132 mutation.

3. The method of embodiment 2, wherein the IDHl R132 mutation is selected from the group consisting of R132C, R132H, R132S, R132G, and R132L.

4. The method of embodiment 1, wherein the detecting an IDHl mutation comprises detecting a single nucleotide variant (SNV) coding the IDHl mutation, wherein the IDHl mutation is selected from the group consisting of R132C, R132H, R132G, R132S, and R132L.

5. The method of embodiment 4, wherein the IDHl mutation is detected using polymerase chain reaction (PCR) technology with homogeneous real-time fluorescence detection.

6. The method of embodiment 1, wherein the detecting an IDHl mutation comprises using an in vitro PCR assay for the qualitative detection of single nucleotide variants (SNVs) coding an IDHl R132 mutation selected from the group consisting of R132C, R132H, R132G, R132S, and R132L in the DNA from the sample.

7. The method of embodiment 1, wherein the sample is obtained from patient bone marrow.

8. The method of embodiment 1, wherein the sample is obtained from patient blood.

9. The method of embodiment 1, wherein the pharmaceutically acceptable composition is formulated for oral administration.

10. The method of embodiment 9, wherein the pharmaceutically acceptable composition is a capsule.

11. The method of embodiment 5, wherein the method further comprises combining the extracted DNA sample with oligonucleotide primers designed to specifically amplify (i) R132C and R132H mutations or (ii) R132G, R132S, and R132L mutations.

12. The method of embodiment 11, wherein the real-time fluorescence signal of each IDHl mutation of either (i) R132C and R132H or (ii) R132G, R132S, and R132L is distinguishable in a single well. 13. A method of treating acute myeloid leukemia (AML) in patients with an isocitrate dehydrogenase- 1 (IDH1) mutation, the method comprising administering twice daily to a patient with an IDH1 mutation 150 mg of olutasidenib in a pharmaceutically acceptable composition, wherein the IDH1 mutation has been detected using an FDA-approved diagnostic test.

14. The method of embodiment 13, wherein the FDA-approved diagnostic test is the IDH1 Assay of Example 14.

15. The method of embodiment 13, wherein the patient is receiving or has received therapy comprising azacitidine or cytarabine.

16. The method of embodiment 13, wherein the AML is relapsed or refractory.

17. A method of treating AML in patients with an isocitrate dehydrogenase- 1 (IDHl) mutation, the method comprising steps of:

determining whether the patient has an IDHl mutation by:

i. obtaining a sample from the patient; and

ii. performing an assay on the sample to determine if the patient has an IDHl mutation; and if the patient has an IDHl mutation, then administering to the patient with the IDHl mutation a total of 150 mg of olutasidenib twice daily in a pharmaceutically acceptable composition, and

if the patient does not have an IDHl mutation, then not administering to the patient with the IDHl mutation a total of 150 mg of olutasidenib twice daily in a pharmaceutically acceptable composition.

18. The method of embodiment 17, wherein the IDHl mutation is an IDHl R132 mutation.

19. The method of embodiment 18, wherein the IDHl R132 mutation is selected from the group consisting of R132C, R132H, R132S, R132G, and R132L.

20. The method of embodiment 17, wherein the assay is the IDHl Assay of Example 14.

EXAMPLES

Example 1 : Compound 1 potently and selectively inhibited 2-HG production in IDH-1 R132H and IDH-1 R132C mutant enzymes in biochemical assays, compared to wild type IDH-1 enzyme and mutant IDH-2 enzymes.

[0218] The biochemical potencies of Compound 1 against IDH-1 R132H and IDH-1 R132C mutants were determined in diaphorase-coupled assays, which measure activity by the determination of the level of remaining co-substrate NADPH after the enzymatic reaction is quenched (Figure 2A and Figure 2B). Figure 2A and Figure 2B are schematics illustrating the working principle of the diaphorase-coupled assay for measuring potency and selectivity of Compound 1 for IDH-1 and IDH-2 enzymes. Recombinant homodimeric IDH-1 R132H or IDH- 1 R132C mutant enzymes were used in these assays.

[0219] Results are shown in Table 6, relative to the ICso value obtained for R132H IDH-1 mutated enzyme. Referring to data in Table 6, Compound 1 was found to selectively inhibit the enzymatic activity of the IDH-1 R132H and IDH-1 R132C mutations with an ICso value within a factor of about 5 (i.e., the ICso value measured for IDH-1 R132C mutant enzyme was about 5 times higher than the ICso measured in the IDH-1 R132H mutated enzyme). The selectivity of Compound 1 against other IDH isozymes was also tested utilizing diaphorase coupled assays employing either wild-type IDH-1 or one of 2 alternate mutated forms of IDH-2, namely IDH-2

R172K and IDH-2 R140.

Table 6

[0220] Compound 1 had comparatively very weak activity against wild type IDH-1 (ICso value of about 922 times greater than the ICso value measured for IDH-1 R132H). Compound 1 also demonstrated very weak activity against IDH-2 R172K that was more than 1,000 greater than the ICso value measured for IDH-1 R132H. Compound 1 did not show any inhibition of IDH-2 R140Q up to a concentration of 100 mM. These selectivity data indicate that Compound 1 is a potent and selective inhibitor of IDH-1 R132 mutations.

Example 2: Compound 1 exhibited specific binding to a surface containing immobilized IDH-1 R132 mutant protein (compared to a comparator surface with immobilized BCL6), with two binding sites having different Kd values detected by Surface Plasmon Resonance analysis.

[0221] The biophysical interaction between Compound 1 and IDH-1 R132H was further characterized using Surface Plasmon Resonance (SPR) technology. Compound 1 was shown to exhibit specific binding to the surface containing immobilized IDH-1 R132H mutant protein compared to a control surface on which the unrelated protein BCL6 was immobilized, where no binding was observed (Figure 3 A and Figure 3B, respectively). Analysis of the SPR data revealed two binding sites between Compound 1 and IDH-1 R132H, with Kd values of 31 nM (with konl = 2.04 ±0.03 X 105 M-ls-1 and koffl= 0.0063 ± 0.0001 s-1) and 1200 nM (with kon2 = 1.56 ±0.03 X 105 M-ls-1 and koffl= 0.187 ± 0.001 s-1 ), respectively. It is likely that the apparent low affinity binding site is an artifact of the immobilization of the protein on the surface of the chip, and as the Kd value for the high affinity binding site is close to the enzymatic ICso of Compound 1 for IDH-1 R132H, this was used to confirm specific binding of Compound 1 to IDH-1 R132H.

Example 3: Compound 1 potently inhibited 2-HG production in IDH-1 R132G, IDH-1 R132L, and IDH-1 R132S mutant cell lines in cell based assays, with ICso values greater than IDH-1 R132C mutant cell lines.

[0222] The cellular potency of Compound 1 in suppressing intracellular 2-HG levels was determined in cell lines expressing five different mutated IDH-1 proteins found in human cancers (R132H, R132C, R132G, R132L, R132S). The human fibrosarcoma cell line HT-1080 harbors a naturally occurring heterozygous IDH-1 R132C mutation. The human colorectal carcinoma cell line HCT 116 is wild type for IDH-1, but heterozygous mutations coding for IDH-1 R132H or R132C were introduced by knock-in into the endogenous IDH-1 gene locus. Finally, the human astrocytoma cell line U-87 MG is also wild type for IDH-1, but expression of five different mutated IDH-1 proteins was achieved by stable transfection.

[0223] The parental HCT116 line (colon) line does not produce high levels of 2-HG, but the variants used herein (X-MAN HCT-116 lines obtained from Horizon Discovery Ltd.) are engineered to knock-in a heterozygous mutation of either IDH-1 R132H or IDH-1 R132C. This recapitulates the cellular context in mIDH-1 cancer cells where there are both wild type and mutant IDH-1 subunits that together form a heterodimer that is responsible for the production of elevated levels of 2-HG. These modified lines can be used as models of IDH-1 mutant disease.

[0224] Each of these cell lines was treated with Compound 1 for 24 hr, and intracellular 2- HG levels were determined by mass spectroscopy. As shown in the Table 7, Compound 1 suppressed 2-HG production in each cell line, with ICso values ranging from less than 10 nM to less than 150 nM. Compound 1 is therefore a potent inhibitor of a variety of clinically relevant IDH-1 mutations in a cellular context. Table 7 shows the ICso values measured relative to the ICso value obtained for U-87 MG/IDH-1 R132G.

Table 7

*Mean ±/- SEM where applicable

Example 4: Testing Compound 1 in mouse xenograft models using HCT 116 cells with R132 C and R132H mutations.

[0225] In order to assess the in vivo activity of Compound 1, PK-PD experiments in mice bearing HCT-116 xenografts (derived from Horizon Discovery isogenic cell lines harboring IDH1-R132H and IDH1-R132C knock-in mutations) were used to determine the degree of exposure required to suppress 2-HG levels. Compound 1 was administered to HCT116-IDHl- R132H/+ xenograft bearing female BALB/c Nude mice at three oral doses (12.5, 25, and 50 mg/kg) in 12-hour intervals. Plasma and xenograft tumor samples were collected at 4, 12, and 24 hours post last dose to determine the exposure of Compound 1 in plasma and tumor, as well as to measure the inhibition of IDH1 mutant activity in tumor based on the reduction in levels of 2-HG. In IDH1-R132H/+ xenograft models, the free concentration of Compound 1 was comparable in plasma and xenograft tumors, and exposures were dose-dependent (Figure 4A and Figure 4B). In comparison to the vehicle treated group, Compound 1 showed a time and dose- dependent inhibition of 2-HG levels in plasma and in tumor (Figure 4C). At the highest dose tested in these studies (50 mg/kg), treatment with Compound 1 inhibited 2-HG levels in the tumor by >90% for up to 24 hours after the last dose in the HCT116-IDH1-R132H/+ xenograft model, and to similar levels for at least 12 hours in the HCT116-IDH1-R132C/+ model.

Calculations based upon the percentage of suppression of 2-HG concentration in tumor versus the free drug concentration in tumor gave in vivo IC50 values of 26 nM and 36 nM in the HCT116-IDH1-R132H or HCT116-IDH1-R132C models, respectively. When corrected for unbound levels of Compound 1, there is an excellent correlation in potency among the biochemical assay, cellular assay, and in vivo studies.

Table 8

[0226] In order to optimize the dosing schedule of Compound 1 to achieve sustained >90% 2-HG inhibition in mIDH-1 in vivo, HCT116 (IDH-1 R132H)and HCT116 (IDH-1 R132C) xenograft-bearing mice were treated with Compound 1 at 25 and 50 mg/kg BID (3 doses). The free drug concentration of Compound 1 at 12 hour post final dose is above the in vivo IC90 for all doses, and a greater than 90% reduction of 2-HG levels in tumor were achieved in each case. The free drug concentration decreased to 3-10X the in vivo IC50 at 24 hour post final dose, and Compound 1 showed 80-90% inhibition. There was less than 20 nM free drug concentration in tumor at 48 and 72 hours after final dose, and at that point there was less than 50% 2-HG inhibition in tumor samples, consistent with the reduced level of Compound 1.

[0227] Briefly, 5x106 HCT-116 IDH1-R132H/+ cells (Horizon Discovery) in PBS was inoculated subcutaneously at the right flank of the 6 weeks old female BALB/c nude mice. When the tumor size reached 360-400 mm3, mice were randomized by tumor volume into nine mice per group. The tumor bearing mice were treated with vehicle (9: 1 PEG400:Ethanol) or

Compound 1 for three doses with 12 hr dosing interval. The dosing volume was 10 pL/g. The plasma samples and tumor samples were collected at 4, 12 and 24 hr post final dose (N=3 mice per time point) for the subsequent measurement of compound level in plasma and tumor samples and of 2-HG level in the tumor samples by UPLC-MS-MS system.

[0228] In a separate dosing example, 5x106 HCT-116 IDH1-R132C/+ cells (Horizon Discovery) in PBS was inoculated subcutaneously at the right flank of the 6-8 weeks old female BALB/c nude mice. When the tumor size reached -250 mm3, mice were randomized by tumor volume into nine mice per group. The tumor bearing mice were treated with vehicle (9: 1 PEG400:Ethanol) or Compound 1 for six doses with 12 hr dosing interval. The dosing volume was 10 pL/g. The plasma samples and tumor samples were collected at 4, 8 and 12 hr post final dose (N=4 mice per time point) for the subsequent measurement of compound level in plasma and tumor samples and of 2-HG level in the tumor samples by UPLC-MS-MS system.

[0229] For each assay, the total concentration of Compound 1 was determined in mM and free Compound 1 concentration was calculated by multiplying the total Compound 1

concentration by 0.043 given that Compound 1 is 95.7% protein binding in mouse plasma. The percentage of 2-HG inhibition in individual tumor sample in the treated groups was normalized to the average of 2-HG concentration in the vehicle group at the corresponding sampling time using the following calculation: % 2-HG inhibition = 100*(A-B)/A, where A is the average of 2- HG concentration at the corresponding sampling time, B is the 2-HG concentration in the tumor treated with given dose of Compound 1 and sacked at the given sampling time. The in vivo potency of Compound 1 for suppressing 2-HG in tumor is calculated by plotting the percentage of 2-HG inhibition against corresponding free Compound 1 concentration in tumor and fitting the data with four-parameter logistic equation.

IDH1-R132H mutation

[0230] IDH1-R132H mutation resulted in elevation of 2-HG level in hematological and solid cancers. HCT-116 IDH1-R132H/+ xenograft tumor was used to assess the in vivo potency of Compound 1 to suppress 2-HG in tumor lysates. The tumor bearing mice were randomized by tumor size into twelve mice per group. The mice were treated with Compound 1 at 6.25, 12.5, 25, or 50 mg/kg for six doses with dose interval of 12 hr. The plasma and tumor samples were collected at 4, 8, and 12 hr post last dose with four mice per time point. The Compound 1 concentration in plasma and tumor samples was analyzed by LC-MS method. The 2-HG level in tumor samples was analyzed by LC-MS method. The percentage of 2-HG suppression in tumor lysate at given dose of Compound 1 was then normalized to 2-HG level in the vehicle control group. The dose and time dependent 2-HG inhibition by Compound 1 was observed in this study. The degree of 2-HG inhibition in tumor lysates was correlated with the free drug concentration in the corresponding tumor lysate. The calculated in vivo potency of Compound 1 to suppress 2- HG in tumor was 26.0 nM.

[0231] Upon correcting for unbound Compound 1 concentration, there was a good correlation between the enzymatic, cellular 2-HG, and in vivo 2-HG potencies of Compound 1 for IDH1-R132H mutant. IDH1-R132C mutation

[0232] IDH1-R132C mutation resulted in elevation of 2-HG level in hematological and solid cancers. HCT-116 IDH1-R132C/+ xenograft tumor was used to assess the in vivo potency of Compound 1 to suppress 2-HG in tumor lysates. The tumor bearing mice were randomized by tumor size into nine mice per group. The mice were treated with Compound 1 at 12.5, 25, or 50 mg/kg for three doses with dose interval of 12 hr. The plasma and tumor samples were collected at 4, 12, and 24 hr post last dose with three mice per time point. The Compound 1 concentration in plasma and tumor samples was analyzed by LC-MS method. The 2-HG level in tumor samples was analyzed by LC-MS method. The percentage of 2-HG suppression in tumor lysate at given dose of Compound 1 was then normalized to 2-HG level in the vehicle control group. The dose and time dependent 2-HG inhibition by Compound 1 was observed in this study. The degree of 2-HG inhibition in tumor lysates was correlated with the free drug concentration in the corresponding tumor lysate. The calculated in vivo potency of Compound 1 to suppress 2-HG in tumor was 36.0 nM.

[0233] Upon correcting for unbound Compound 1 concentration, there was a good correlation between the enzymatic, cellular 2-HG, and in vivo 2-HG potencies of Compound 1 for IDH1-R132C mutant.

Results

[0234] Given the role of 2-HG in suppressing normal differentiation of mt-IDHl cells (Figueria et al., 2010; Saha et al., 2014), it is hypothesized that in order to reverse and maintain this effect, it is necessary to achieve a very high degree of target inhibition on a continuous basis. Therefore, in order to optimize the dosing schedule of Compound 1, it was necessary to achieve sustained >90% 2-HG inhibition in mt-IDHl in vivo. For the HCT116-IDH1R132H xenograft assay, the 12 and 24 hour time points were chosen to reflect the compound level and

corresponding 2-HG inhibition at the Ctrough of BID and QD dosing schedules. The 48 and 72 hour time points were selected to investigate whether Compound 1 had long lasting effects on 2- HG inhibition. The free drug concentration of Compound 1 at 12 hour post final dose is above the in vivo IC90 for all doses, and a greater than 90% reduction of 2-HG levels in tumor were achieved in each case. The free drug concentration decreased to 3-10X the in vivo IC50 at 24 hour post final dose, and the compound showed 80-90% inhibition. There was less than 20 nM free drug concentration in tumor at 48 and 72 hours after final dose, and at that point there was less than 50% 2-HG inhibition in tumor samples, consistent with the reduced level of Compound 1. These data support the premise that constant target coverage by a significant margin is required to achieve sustained 2-HG inhibition. This experiment also suggests that a BID schedule is the preferred dosing regimen for Compound 1 in order to continuously achieve >90% 2-HG inhibition. This level of inhibition has recently been correlated to clinical efficacy with AG-221 in mt-IDH2 harboring AML patients (Fan et al., 2014)).

[0235] The present disclosure contemplates, among other things, recognition that the total concentration (Ceff) of Compound 1 must be above 1652 ng/mL in human patients in order to achieve 90% inhibition of 2-HG and above 2000 ng/mL to achieve greater than 90% inhibition of 2-HG. Ceff was determined using assays outlined in this Example. In two separate mouse experiments, HCT-116 IDH1-R132H/+ xenografts and HCT-116 IDH1-R132C/+ xenograft tumor were used to assess the in vivo potency of Compound 1 to suppress 2-HG in tumor lysates. Compound 1 concentration in plasma and tumor samples and 2-HG level in tumor samples was measured. The degree of 2-HG inhibition in tumor lysates was correlated with the free drug concentration in the corresponding tumor lysate (see Figure 4D). Given the role of 2-HG in suppressing normal differentiation of mt-IDHl cells (Figueria et al., 2010; Saha et al., 2014), the present disclosure hypothesized that in order to reverse and maintain this effect, it is necessary to achieve a very high degree of target inhibition with Compound 1 on a continuous basis. It was previously proposed that >90% inhibition of 2-HG correlates to clinical efficacy in mt-IDH2 harboring AML patients (FAN, B. et al., Evaluation of the pharmacokinetic/pharmocodynamic (PK/PD) relationships of an oral, selective, first-in-class, potent IDHl inhibitor, AG-221, from a phase 1 trial in patients with advanced IDH2 mutant positive hematologic malignancies, Blood, 124: 3737, 6 pages (2014)). Using the curve from Figure 4D, the level of free drug concentration of Compound 1 was determined to be 0.256 mM in order to achieve 90% inhibition of 2-HG.

[0236] Using a rapid equilibrium dialysis approach, the plasma protein binding for a human patient was determined to be 94.5%. (Waters, N.J., et al. (2008)). Validation of a rapid equilibrium dialysis approach for the measurement of plasma protein binding. (J Pharm Sci 97(10): 4586-95.) Accordingly, the total concentration (Ceff) can be determined: 0.256/((100- 94.5)/100) =4.65 mM=1652 ng/mL. Example 5: Pharmaceutical Compositions in an Oral Dosage Form of Compound 1

[0237] A therapeutically effective amount of Compound 1 can be orally administered (e.g., an amount providing a steady state blood concentration greater than the IC90 for 2-HG

production for cancer cells having the IDH-1 R132 mutation disclosed herein, and less than an amount of about 7,200 ng/mL). For example, a therapeutically effective amount of Compound 1 can provide a steady state blood concentration of about 2,000 ng/mL to 7,200 ng/mL throughout the course of treatment. The therapeutically effective amount can be up to about 150 mg of Compound 1 in the solid form obtained by the method of Example 5, administered to the patient BID on consecutive days, e.g., throughout a course of treatment of at least about 6 months.

[0238] Step 1: Compound 1 can be obtained using the chemical synthesis disclosed in PCT patent application publication WO2016/044789A1 (published March 24, 2016; filed September 18, 2015) and summarized in Figure 5. Examples 1, 21 and 25 from WO2016/044789A1 are incorporated herein by reference, along with associated analytical methods disclosed in the publication WO2016/044789A1. Briefly, Compound 1 can be obtained using the method of Example 25 (pages 92-93), based on the reaction of Intermediate II-l (obtainable using the method of Example 1 on pages 26-27) and Intermediate III-l (obtainable using the method of Example 21 on pages 79-82). Using this method, Compound 1 was obtained as a white solid (790 mg) m.p. 262-264 °C. ¾ NMR (300 MHz, DMSO- de) d: 12.07 (s, 1H), 7.75 (s, 1H), 7.73 (d, J= 2.2 Hz, 1H), 7.51 (dd, J= 8.6, 2.3 Hz, 1H), 7.31 (d, J = 8.8 Hz, 1H), 6.97 (d, J = 8.0 Hz, 1H), 6.93 (d, J = 7.7 Hz, 1H), 5.95 (d, J = 8.0 Hz, 1H), 4.68 (m, 1H), 3.58 (s, 3H), 1.50 (d, J =

6.6 Hz, 3H). LCMS (Method 3): 100% pure @ 254 nm, Rt 10.78 min, m z 355, 357 [M+H]+.

The filtrate and the colored fractions (TLC pure) from the second ISCO were combined and treated with activated charcoal and filtered (until the filtrate is colorless). The filtrate was then concentrated under reduced pressure on rotavap to remove dichlorometane until a lot of white solid precipitated out. The white solid was collected by filtration and washed with cold MeOH. It was then mixed with MeCN/H20 (10 mL/25 mL) and lyophilized to afford the title compound 1- 13 as a white solid (970 mg) m.p. 262-264 °C. ¾ NMR (300 MHz, DMSO-d6) d: 12.06 (s, 1H), 7.75 (s, 1H), 7.73 (d, J = 2.5 Hz, 1H), 7.51 (dd, J = 8.6, 2.3 Hz, 1H), 7.31 (d, J = 8.8 Hz, 1H),

6.97 (d, J = 8.0 Hz, 1H), 6.92 (d, J= 8.0 Hz, 1H), 5.95 (d, J= 8.0 Hz, 1H), 4.68 (m, 1H), 3.58 (s, 3H), 1.50 (d, J = 6.9 Hz, 3H). LCMS (Method 3): 100% pure @ 254 nm, m/z 355, 357 [M+H]⁺. The total yield for combined two batches is >67%. [0239] Step 2: Next, a solid form of Compound 1 can be obtained that is useful in an oral dosage form. Unless otherwise indicated, the studies herein were performed using a pharmaceutically acceptable solid form in an oral dosage form of Compound 1 that can be obtained by the method of Step 2 of Example 5. All volumes are with respect to the quantity of Compound 1 (v/w). Compound 1 obtained from Step 1 above is dissolved in 18 volumes of dichloromethane. The resulting solution is then concentrated under reduced pressure to approximately 5 volumes. To the mixture is added 5 volumes of ethyl acetate. The mixture is concentrated under reduced pressure to 5 volumes. To the mixture is added an additional 5 volumes of ethyl acetate, and the mixture again concentrated under reduced pressure to 5 volumes. The mixture is diluted to 10 volumes with ethyl acetate, and the mixture stirred at room temperature for 18 hours and then cooled to 0 °C. The mixture is stirred at 0 °C for 3 hours and then filtered. The solids are rinsed with ethyl acetate and dried under vacuum (counterbalanced by nitrogen) at ambient temperature.

[0240] Step 3: The oral dosage form of Compound l is a pharmaceutically acceptable solid form of Compound 1 and can be obtained using the method of Example 5 Step 2. The oral dosage form does not contain associated solvent or a counterion. In particular, the oral dosage form of Compound 1 can be a capsule comprising drug substance (Compound 1) blended with excipients to improve powder flow and encapsulated in a Coni-Snap® hard gelatin capsule suitable for oral dosage in humans.

[0241] A pharmaceutically acceptable solid form of Compound 1 can be identified using reflection X-ray powder diffraction (XRPD) pattern of Compound 1. High resolution X-ray Powder Diffraction experiments can be performed with Panalytical X’Pert3 Powder XRPD on a Si zero-background holder. The 2 theta position can be calibrated against Panalytical 640 Si powder standard. Details of the XRPD method are listed below in Table 9, with XRPD peaks reported as diffraction angles at 2 theta, with d-spacing measured in angstroms.

Table 9

[0242] An example of a pharmaceutically acceptable solid form of Compound l is a solid form characterized by a reflection X-ray powder diffraction (XRPD) pattern comprising characteristic peaks at 6.3, 12.8, 13.8, 23.6, and 27.8 degrees ±0.2° 2Q. A pharmaceutically acceptable solid form of Compound l is a solid form characterized by characterized by an X-ray Powder Diffraction (XRPD), having diffractions at angles (2 theta ±0.2) of 6.3, 12.8, 13.8, 23.6, and 27.8, corresponding to d-spacing (angstroms ±0.2) of 14.0, 6.9, 6.4, 3.8, and 3.2, respectively. In some embodiments, a pharmaceutically acceptable solid form of Compound 1 can be identified by X-ray Powder Diffraction (XRPD), having characteristic diffractions at angles (2 theta ± 0.2) of 5.7, 6.3, 8.5, 10.6, 12.8, 13.8, 17.3, 22.0, 22.8, 23.6, and 27.8. In some embodiments, a pharmaceutically acceptable solid form of Compound 1 can be identified by X- ray Powder Diffraction (XRPD), having characteristic diffractions at angles (2 theta ± 0.2) of 5.7, 6.3, 8.5, 10.6, 12.8, 13.8, 17.3, 22.0, 22.8, 23.6, and 27.8, corresponding to d-spacing (angstroms ± 0.2) of 15.4, 14.0, 8.4, 6.9, 6.4, 5.1, 4.0, 3.9, 3.8, and 3.2, respectively.

Example 6: Comparative compounds demonstrated greater disparity between 2-HG inhibition in R132C and R132H IDH-1 cells, compared to Compound 1.

[0243] The comparative activity of each of a series of mIDH-1 inhibitor compounds including Compound 1 were measured using the cell based assay in Example 3. The ratio of the IC50 values obtained from IDH-1 R132C HCT116 mutant cells (IC50 mM g mean) / IC50 values obtained from IDH-1 R132H HCT116 mutant cells (IC50 pM g mean) is provided in Table 10. Compound 1 had the lowest ratio among the tested compounds, indicating near equipotent activity of Compound 1 as measured with the R132C and R132H IDH-1 mutant cell assay of Example 3 (using the HCT 116 cells described in Example 3). Compound 1 showed

comparative activity inhibiting 2-HG production from m IDH-1 R132C and R132H cell lines (using the assay of Example 3) that was within 5-fold, compared to more disparate differences in activity ranging from about 8-fold to over 200 fold (240) in comparative compound A-H in Table 10. Table 10

Example 7: Determination of central nervous system multiparameter optimization (CNS MPO)

[0244] Central nervous system multiparameter optimization (CNS MPO) may be used to prioritize compounds based on their likelihood to be brain-penetrant. The scoring function uses six key physicochemical properties (scoring each parameter on a scale of zero to one) to arrive at a composite score ranging from 0-6. Higher CNS MPO scores are correlated with a higher likelihood of a compound being brain-penetrant. The reported CNS MPO scores were calculated following the method reported in: Wager, T. T., Hou, X., Verhoest, P. R., and Villalobos, A. (2010) Moving beyond rules: The development of a central nervous system multiparameter optimization (CNS MPO) approach to enable alignment of druglike properties. ACS Chem. Neurosci. 7, 435-449.

[0245] A summary of the MPO scores for several mIDH inhibitors are found in Table 11 : Table 11

*For Compound 1 and Compounds 1-1 through 1-29, WO2016/044789 defines ICso values for

IDH1 R132H as“++++”: < 0.01 mM;“+++”: between 0.01 mM and 0.1 pM;“++”: from 0.1 pM to 1 pM; and ICso values for IDH1 R132C as“++++”:< 0.1 pM;“+++”: between 0.1 pM and 1 pM;“++”: from 1 pM to 10 pM; and“+”: > 10 pM

a. CNS MPO score calculated based on the method described in T. Wager, ACS Chem. Neurosci. (2010), 1, 435-449.

b. Literature reported data

c. Predicted BBB+: CNS MPO score >3.8

d. WO2016/044789 also reports Compound 1 (as 1-13) as having activity in HCT116 mutant IDH1 R132H and R132C cells as +++ and +++, respectively.

e. WO2016/044789 also reports 1-25 as having activity in HCT116 mutant IDHl R132H and R132C cells as ++++ and ++++, respectively.

1. Popovici-Muller, L, et al. Discovery of AG-120 (Ivosidenib): A First-in-Class Mutant IDHl Inhibitor for the Treatment of IDHl Mutant Cancers. ACS Med. Chem. Lett., 2018, 9(4), 300- 305.

2. Yen, K. , et al. Abstract B126: AG-881, a brain penetrant, potent, pan-mutant IDH (mIDH) inhibitor for use in mIDH solid and hematologic malignancies, AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; October 26-30, 2017; Philadelphia, PA.

3. Cho, Y. S. , et al. Discovery and evaluation of clinical candidate IDH305, a brain penetrant mutant IDH1 Inhibitor. ACS Med. Chem. Lett. 2017, 8 , 1116-1121.

4. Levell, J. R. , et al. Optimization of 3-pyrimidin-4-yl-oxazolidin-2-ones as allosteric and mutant specific inhibitors of IDHl. ACS Med. Chem. Lett. 2017, 5, 151-156.

5. Okoye-Okafor, U. C., et al. New IDHl mutant inhibitors for treatment of acute myeloid leukemia. Nat. Chem. Biol. 2015, 77, 878-886.

6. Pusch, S. , et al. Pan-mutant IDHl inhibitor BAY 1436032 for effective treatment of IDHl mutant astrocytoma in vivo. Acta Neuropathologica 2017, 733(4), 629-644.

Example 8: Determination of Predicted C_bram Ratio of mIDH inhibitors

[0246] The distribution of Compound 1, AG-120 or AG-881 into the brain was measured ex- vivo, individually, in Sprague Dawley rats (n=4/molecule) following a 6 hour intravenous infusion of a 7.5 mg/kg dose. Brain tissue and plasma samples were collected at the end of the infusion time and were processed for bioanalysis via tandem HPLC-mass spectrometry analysis (LCMS) to determine the total amount of compound present. In parallel, brain and plasma samples were subjected to equilibrium dialysis as described by N.J. Waters et.al (J. Pharm. Sci. (2008) 97(10):4586-95) to determine the unbound fraction (Table 12). The brain distribution or partitioning coefficient (Kpuu) was then calculated as the ratio of the unbound concentration of drug in brain (Fu, brain) to the unbound concentration of drug in plasma (Fu, plasma). Similarly, all calculations of effective plasma concentrations in rodents or humans were conducted using the unbound/free fraction measured for each molecule in each species using conventional equilibrium dialysis as described by Waters.

[0247] To determine the projected effective free concentration in humans, the plasma concentrations in tumor bearing mice were measured using 90% inhibition in tumors (IC90) of the biomarker 2-HG, as the minimal effective concentration to provide therapeutic benefit. In tumor bearing mice studies, the unbound plasma concentration at IC90 was determined to be 90.7 ng/mL (Table 12). In human clinical trials, a dose of 150 mg BID for Compound 1 showed a mean plasma concentration (C&ee, avg) of 171 ng/mL, which when corrected by the expected brain partitioning (Kpuu = 0.42) and free fraction in brain (Fu, brain =0.05) provides an estimate of 3.6 ng/mL unbound concentration in brain. The target effective concentration in human brain based on mouse models (to achieve IC90) is 1.91 ng/mL. Hence, Compound 1 partitions into the brain by 2-fold greater than projected levels required to achieve therapeutic benefit.

Table 12

Calculated brain distribution of Ceff,fu plasma in mice

**Based on Cycle 1 PK, 2-fold decrease over time

Example 9 - Compound 1 penetrates the blood brain barrier in murine models

[0248] The blood brain barrier penetration and free brain exposure of Compound 1 was investigated in the male CD-I mouse (Figure 6). The brain exposure of Compound 1 following oral dosing (5 mg/kg) was evaluated. Following a 5 mg/kg oral dose in CD1 mice, the brain to plasma ratio of Compound 1 was found to be 0.24 suggesting that the compound has reasonable brain penetration characteristics, and that at higher doses Compound 1 would have the potential to achieve therapeutic brain levels.

[0249] This was confirmed by dosing at 100 mg/kg (Figure 7), where the brain to plasma ratio was determined to be 0.38. Importantly, following a single 100 mg/kg PO dose, the free Cmax concentration of Compound 1 was about 200 nM (approximately ten fold IC50), dropping to about 130 nM (about six fold IC50) at the 7 hour time point, which is consistent with exposures that produced >90% suppression of 2-HG in both HCT116 (IDH-1 R132H) and HCT116 (IDH-1 R132C) xenograft PK-PD studies. Based on assessment in the mouse, Compound 1 crosses the blood-brain barrier to reach free concentration levels in the brain consistent with

pharmacological activity. An oral 100 mg/kg dose gives a free Cmax in brain of 200 nM, dropping to about 130 nM at the 7 hr time point. Example 10: Testing Compound 1 in cynomolgus monkey models.

Animal Acquisition and Acclimation

[0250] A total of 22 male and 22 female experimentally naive cynomolgus monkeys, approximately 2 years and 7 months to 3 years and 11 months of age at transfer, were transferred from the stock colony. The animals were originally received from Worldwide Primates Inc. Animals were quarantined upon arrival and quarantine activities, including intrapalpebral tuberculin skin tests, were performed. The animals were considered suitable prior to being released from quarantine. During acclimation as part of the stock colony, the monkeys were examined by a staff veterinarian, weighed, and observed daily with respect to general health and any signs of disease.

Randomization, Assignment to Study, and Maintenance

[0251] Using a standard, by weight, measured value randomization procedure, 20 male and 20 female animals (weighing 2.50 to 3.15 kg and 2.35 to 3.20 kg, respectively, at randomization) were assigned to the control and 3 treatment groups. Animals assigned to study had body weights within ±20% of the mean body weight for each sex. Extra animals obtained for the study, but not placed on study, were transferred to the stock colony. Each animal was assigned an animal number to be used in the data collection system and was implanted with a microchip bearing a unique identification number. Each animal was also identified by a permanent tattoo with the vendor animal number. The individual animal number, implant number, tattoo number and study number comprised a unique identification for each animal. Each cage was identified by the animal number, study number, group number, and sex.

Administration

[0252] Male and female cynomolgus monkeys were assigned to four groups. Animals were assigned to the study as indicated below in Table 13. Six animals per sex in Group 1 were administered vehicle control article only. Four animals per sex in Groups 2 and 3, and 6 animals per sex in Group 4 were administered test article. Animals in Groups 2 through 4 were dosed for 28 days. 2 animals/sex in Groups 1 & 4 were assigned for 28-day recovery assessment. Animals were dosed via oral gavage twice daily, every 12 hours, at a volume of 10 mL/kg/dose

(20 mL/kg/day). Animals in Groups two through four were administered 30, 100/50, or

300/200/100 mg/kg/day (15, 50/25, or 150/100/50 mg/kg/dose) respectively. The dose levels were lowered for Group 3 and Group 4 animals based clinical observations during the dosing phase. The vehicle control article was Kolliphor EL:Tween 80 (70:30, v/v). Animals designated for recovery sacrifice (2 animals/sex in Groups 1&4) underwent 4 weeks of recovery assessment.

Table 13

a Animals in Group 1 received the vehicle, Kolliphor EL:Tween 80 (70:30, v/v).

b Two animals/sex were maintained for a 28-day recovery period.

c Beginning on Day 14, the 50 mg/kg/dose twice daily (BID) dose (100 mg/kg/day) was reduced to 50 mg/kg/dose once daily (50 mg/kg/day) for the remainder of the study.

d Animals at 300 mg/kg/day were placed on a dosing holiday beginning with the second dose on Days 4 through Day 11. The 150 mg/kg/dose twice daily (BID) dose (300 mg/kg/day) was reduced to 100 mg/kg/dose twice daily (200 mg/kg/day) at a concentration of 10 mg/mL for Days 12 and 13. Beginning on Day 14, the 100 mg/kg/dose twice daily (BID) dose (200 mg/kg/day) was reduced to 100 mg/kg/dose once daily (100 mg/kg/day) for the remainder of the study.

[0253] Assessment of toxicity was based on mortality, clinical observations, body weights, food consumption, ophthalmic observations, electrocardiographic (ECG) measurements, and clinical and anatomic pathology. Blood samples were collected for toxicokinetic evaluations. Electrocardiographic Assessment

[0254] With the exception of one animal in the high dose group, there was no effect of Compound 1 on qualitative ECG parameters. Frequent ventricular premature complexes (Day 1, 3-4 hours post-dose) and ventricular tachycardia (Day 28 pre-dose and 3-4 hours post-dose) were observed in one animal following administration of the 300/200/100 mg/kg/day dose. As these ventricular arrhythmias are not considered normal variants and were observed following the high dose, these findings may have been test article-related. Noteworthy effects on quantitative ECG parameters were limited to QTc interval duration. When evaluated statistically by sex, mean QTc interval duration was longer (vs. concurrent vehicle control value) in 300/200/100 mg/kg/day males at the Day 1 post-dose interval and at the Day 28 pre-dose and post-dose intervals and in 100/50 mg/kg/day females at the pre-test and Day 28 intervals. The difference in 100/50 mg/kg/day females was not considered to be test article-related as it was present prior to initiation of test article administration. In 300/200/100 mg/kg/day males, the increase in mean QTc interval duration may have been test article-related as it was observed at the highest dose level and exhibited a progressive increase with continued dosing. The magnitude of change from pretest values was mild to moderate (Day 1 post-dose: 7.14%; Day 28 pre-dose: 7.73%; Day 28 post-dose: 10.6%). Compared to pretest values, the magnitude of the increase in QTc interval duration in males at the Day 28 post-dose interval was 10.61%, which approximates the 10% change seen in the Japanese QT PRODACT studies (Ando, K., Hombo, T., Kanno, A., Ikeda, H., et al. QT PRODACT: in vivo QT assay with a conscious monkey for assessment of the potential for drug induced QT interval prolongation. J Pharmacol Sci. 2005; 99(5):487-500) of drugs known to cause QT prolongation in people. The effect on QTc interval duration was reversible, not being present at the recovery interval. The threshold Cmax plasma concentration in a monkey that experienced an average group QT_C prolongation of 10.6% was 7840 ng/mL.

Conclusion

[0255] Cynomolgus monkeys tolerated oral doses of Compound 1 twice daily at 15 mg/kg/dose (30 mg/kg/day) and once daily at 50 mg/kg/day. Compound 1-related post-mortem findings at the end of the treatment period included macroscopic findings of black discoloration in the liver of 3/4 males and 2/4 females at 300/200/100 mg/kg/day, which correlated to multinucleated cells in the sinusoids. Lower thymus weights, which correlated to an increased incidence and/or severity of lymphoid depletion, were observed in Compound 1 -treated animals and were considered secondary to stress and ill-health. Microscopically, Compound 1 was associated with multinucleated cells in the sinusoids of the liver and mucosal atrophy of the intestinal tract in males and females at 100/50 and/or 300/200/100 mg/kg/day. The histological changes in the liver were considered adverse and correlated with a number of the serum chemistry changes. At the recovery interval, intestinal changes were absent, suggesting reversibility, and multinucleated cells in the sinusoids of the liver were decreased in severity, indicating partial recovery. Test article-related increase in the mean QT_C level was observed at the highest dose level 300/200/100 mg/kg/day in the male monkeys. The threshold plasma Cmax concentration in a monkey that experienced an average group QT_C prolongation of 10.6% was 7840 ng/mL.

[0256] The no-observed-adverse effect-level (NOAEL) for Compound 1 was considered to be 30 mg/kg/day (15 mg/kg/dose BID). Systemic exposure (Cmax and AUCTlast; combined- sex) at the NOAEL on Day 28 was 2490 ng/mL and 23600 ng h/mL, respectively. Based on the expected reversibility of adverse hepatic findings, 50 mg/kg/day was considered the highest nonseverely toxic dose (HNSTD). Systemic exposure (Cmax and AUCTlast; combined-sex) at the HNSTD on Day 28 was 4350 ng/mL and 53900 ng h/mL, respectively.

Example 11: Compound 1 Induces Mutation Clearance in Patients with Acute Myeloid Leukemia (AML) or Myelodysplastic Syndrome (MDS) Treated in Phase 1 Dose Escalation and Expansion Study

[0257] Isocitrate dehydrogenase 1 mutations (i mIDH-1 ) occur in 7-14% of AML patients (“pts”) and 3% of MDS pts. Compound 1 is a highly potent, selective small molecule inhibitor of mIDH-1 without anticipated CYP or QTc liabilities at the recommended phase 2 dose.

This study evaluated the safety, pharmacokinetics (PK), pharmacodynamics (PD) and clinical activity of the novel anticancer drug Compound 1, administered to patients with relapsed or refractory acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS). Compound 1 is a potent, selective, orally bioavailable, small-molecule inhibitor of mutated isocitrate

dehydrogenase 1 (IDHl) and is intended for the treatment of patients harboring IDHl mutations, in both hematologic and solid tumors.

[0258] The presence of mutations at codon 132 in IDHl imparts a neomorphic activity to the enzyme, resulting in the production of the“oncometabolite” (R)-2-hydroxyglutarate (2-HG), which has pleotropic roles in tumorigenesis. Studies in genetically engineered mouse models and models derived from cancer patient samples support the hypothesis that mutated IDHl produces 2-HG, the downstream effects of which cause epigenetic changes that consequently block the proper differentiation of progenitor cells and lead to cancer. These data support the therapeutic rationale that inhibition of mutated IDHl will lower (R)-2-hydroxyglutarate (2-HG) levels and restore normal cellular differentiation.

Inclusion Criteria

[0259] To be considered eligible to participate in this study, a patient met the inclusion criteria listed below: Pathologically proven AML (except acute promyelocytic leukemia with the t(15; 17) translocation) or intermediate risk, high risk or very high risk MDS as defined by the World Health Organization (WHO) criteria or Revised International Prognostic Scoring System (IPSS-R) harboring IDH1-R132 mutations, and one of the following based on enrollment stage or treatment cohort:

a. Single Agent Phase 1 Cohorts including Dose-Escalation/ Dose-Expansion:

AML/MDS either R/R to standard therapy, or for whom standard treatments are contraindicated

b. Combination (Compound 1 + azacitidine) Phase 1 Dose-Escalation/ Dose-Expansion (patients must meet one of the following):

i. Patients with AML that is either R/R to standard therapy, or for whom standard treatments are contraindicated

ii. Patients with MDS that is either R/R to standard therapy, or are treatmentnaive, who are eligible for azacitidine therapy

c. Combination (Compound 1 + cytarabine) Phase 1 Dose-Escalation/Dose-Expansion Cohort: Patients > 60 years with treatment-naive AML for whom standard treatments are contraindicated

d. Phase 2 Cohort 1 (Single Agent) only: AML R/R to standard therapy

e. Phase 2 Cohort 2 (Single Agent) only: AML in morphologic CR/CRi after prior therapy (+/- HSCT) with residual IDH1-R132 mutation (> 0.01%) detected in the bone marrow

f. Phase 2 Cohort 3 (Single Agent) only: R/R AML/MDS that have been previously treated with IDH1 inhibitor therapy AND for whom standard treatments are contraindicated

g. Phase 2 Cohort 4 (Compound 1 + Azacitidine) only: Patients < 60 years old with R/R AML/MDS with no prior hypomethylating agent therapy AND no prior IDH1 inhibitor therapy

h. Phase 2 Cohort 5 (Compound 1 + Azacitidine) only: R/R AML/MDS that have

inadequately responded to or have progressed on prior treatment with a

hypomethylating agent i. Phase 2 Cohort 6 (Compound 1 + Azacitidine) only: R/R AML/MDS that have been previously treated with a single agent IDHl inhibitor as their last therapy prior to study enrollment

j . Phase 2 Cohort 7 (Single Agent) only: Treatment naive AML patients for whom

standard treatments are contraindicated

k. Phase 2 Cohort 8 (Compound 1 + Azacitidine) only: Treatment naive AML patients who are candidates for azacitidine as a first line treatment

(Note for Phase 2 Cohort 7 and Phase 2 Cohort 8: Treatment naive is defined as no prior treatment for AML. Patients may have received a prior treatment for another hematologic malignancy.)

Patients must have documented IDHl -R132 gene-mutated disease as evaluated by the site

Patients > 18 years old

Eastern Cooperative Oncology Group (ECOG) performance status of 0 to 2

Signed informed consent prior to beginning study and undergoing procedures

No prior solid organ allograft

Acceptable liver function:

a. Bilirubin < 2 times upper limit of normal (ULN) (< 3 times ULN in patients with

Gilbert Syndrome)

b. Aspartate transaminase (AST, also referred to as SGOT), alanine transaminase (ALT, also referred to as SGPT) and alkaline phosphatase (ALP) < 3 times ULN

cceptable renal function:

a. Serum creatinine < 1.5 times ULN or calculated creatinine clearance > 50 mL/min (Cockcroft and Gault 1976)

ecovery from the non-hematologic toxic effects of prior treatment to Grade < 1, or baseline value according to NCI CTCAE classification (excluding infertility, alopecia, or Grade 1 neuropathy)

Baseline QTcF < 450 msec (average of the QTcF values of screening triplicate ECGs) Note: This criterion does not apply to patients with a bundle branch block (BBB); for patients with BBB, a cardiology consult is recommended to ensure that QTcF is not prolonged. 11. Negative serum pregnancy test if female of childbearing potential

12. For fertile men and women, agreement to use highly effective contraceptive methods for the duration of study participation and 90 days after the last dose of study medication

13. Agreement for male patients not to donate sperm and for female patients of childbearing potential not to donate ova during the study and for 90 days after the final dose of study drug

14. Phase 2 Cohorts 1 - 8 (SA and combination) only: Pre-treatment peripheral blood and bone marrow aspirate available for retrospective central confirmation of IDH1-R132 mutation is required. Note: Central confirmation of IDH1-R132 mutation is not required for study enrollment.

Exclusion Criteria

[0260] To be eligible for entry into the study, the patient did not meet any of the exclusion criteria

listed below:

1. Phase 1 Single Agent Dose-escalation/Dose-expansion Cohorts and Phase 2 Cohorts 1, 4, 5, 7 and 8 only: Patients who have been treated with an IDH1 targeted therapy are excluded

2. Phase 2 Single Agent Cohorts 1-3 and 7 only: Patients with IDH2 mutation detection at baseline or history of IDH2m inhibitor treatment are excluded

3. History of prior malignancy unless disease-free for > 12 months or considered surgically cured; patients with nonmelanoma skin cancers or with carcinomas in situ are eligible regardless of the time from diagnosis (including concomitant diagnoses)

4. Patients with symptomatic central nervous system (CNS) metastases or other tumor

location (such as spinal cord compression, other compressive mass, uncontrolled painful lesion, bone fracture, etc.) necessitating an urgent therapeutic intervention, palliative care, surgery or radiation therapy

5. Patients with previous allogeneic HSCT, if they meet any of the following criteria: < 100 days from time of HSCT; active acute or chronic graft vs. host disease (GvHD); or receiving immunosuppressive therapy as treatment or prophylaxis against GvHD Note: Doses < 20 mg methylprednisolone (or its equivalent) daily are not an exclusion criterion. Treatment with radiation therapy, major surgery (requiring general anesthesia) within one month prior to study drug dosing

Treatment with chemotherapy or small molecule anticancer therapeutic within five half- lives of the agent or within 21 days if the half-life is unknown. Patients reenrolling in Cohort 6 after relapse/progression on Cohort 1 are exempt from this washout requirement (i.e. can continue Compound 1 treatment until re-enrollment) 8. Treatment with an anticancer therapeutic antibody less than four weeks before first dose of study drug Treatment with other experimental therapies or participation in another clinical trial within a period of time that is less than the cycle length or within 21 days prior to starting study drug, whichever is shorter

Patients unable to swallow oral medications, or patients with gastrointestinal conditions (e.g., malabsorption, resection, etc.) deemed by the Investigator to jeopardize intestinal absorption

Congestive heart failure (New York Heart Association Class III or IV) or unstable angina pectoris; previous history of myocardial infarction within one year prior to study entry, uncontrolled hypertension, or uncontrolled arrhythmias

Patients with a family history of QT prolongation

Concomitant medication(s) known to cause Torsades de Pointes (TdP) initiated less than the duration required to reach steady-state plasma concentration (approximately five half-lives) before first dose of study drug (medications used as needed [PRN] (e.g. Zofran) are exempt)

Concurrent treatment with chronic corticosteroids except if chronic treatment with < 20 mg of methylprednisolone daily or equivalent (pulse steroids for treatment or prophylaxis are allowed [e.g., for transfusion or medication reactions])

Known HIV positivity

Active, uncontrolled bacterial, viral, or fungal infections, requiring systemic therapy (prophylactic systemic antimicrobials permitted)

Uncontrolled disease-related metabolic disorder (e.g., hypercalcemia)

Pregnant or nursing women or women of childbearing potential not using highly effective contraception; male patients not using highly effective contraception. Note: Women of childbearing potential and men must agree to use highly effective contraception prior to study entry and for the duration of study participation and 90 days after. Should a woman become pregnant or suspect she is pregnant while participating in this study, she should inform her treating physician immediately.

19. Serious nonmalignant disease (e.g., hydronephrosis, liver failure, or other conditions) that could compromise protocol objectives in the opinion of the Investigator and/or the Sponsor

20. Unwillingness or inability to comply with procedures either required in this protocol or considered standard of care

21. Medical, uncontrolled disease-related metabolic disorder, psychiatric, cognitive, or other conditions that may compromise the patient’s ability to understand the patient information, give informed consent, comply with the study protocol, or complete the study

22. History of severe allergic reaction to azacitidine (if patient enrolling into azacitidine combination cohort) or low-dose cytarabine (if patient enrolling into cytarabine combination cohort)

23. Prisoners or patients who are involuntarily incarcerated or are compulsorily detained for treatment of either a psychiatric or physical (e.g. infectious disease) illness. Note: Under certain specific circumstances, a person who has been imprisoned may be included or permitted to continue as a patient, if local regulations permit. Strict conditions apply and approval is required.

Treatment/Intervention Plan

[0261] Compound 1 was administered as a single agent or in combination with azacitidine or cytarabine. Compound 1 was supplied as 50 mg or 150 mg capsules and was administered per the protocol defined frequency and dose level. Azacitidine was administered per site's standard of care. Cytarabine will be administered per site's standard of care.

[0262] The Phase 1 stage of the study was split into 2 distinct parts: a dose escalation part, which utilized an open-label design of Compound 1 (single agent), or Compound 1 + azacitidine (combination agent), or Compound 1 + cytarabine (combination agent) administered via one or more intermittent dosing schedules followed by a dose expansion part. The dose expansion part enrolled patients in up to 5 expansion cohorts, exploring single-agent Compound 1 activity as well as combination activity with azacitidine or cytarabine. Patients received only a single dose of study drug (single-agent arm and combination arm) on Cycle 1 Day 1. Following the completion of the relevant Phase 1 cohorts, Phase 2 began enrollment. Patients were enrolled across 6 different cohorts, examining the effect of Compound 1 (as a single agent) and

Compound 1 with azacitidine (combination) on various AML/MDS disease states. Conditions examined include acute myeloid leukemia (also known as acute myelogenous leukemia) and myelodysplastic syndrome. Figure 8A and Figure 8B summarize certain cohorts from a phase 1 study for mIDHI AML and MDS.

Table 14 - Arms and Interventions of Phase 1 Trial.

[0263] Following the completion of Phase 1, Phase 2 enrollment began. Patients were enrolled across 6 different cohorts, examining the effect of Compound 1 (as a single agent) and Compound 1 + azacitidine (combination) on various AML/MDS disease states. The Phase 2 cohorts are summarized in Table 15 below:

Table 15.

Primary Outcome Measures

[0264] The outcome of the study can be evaluated using the following criteria:

1. Maximum Tolerated Doses (MTDs) or Maximum Evaluated Doses (MEDs) [Phase 1] Time Frame: Within first 4 weeks of treatment.

2. Number of Participants with a Dose Limiting Toxicity (DLT) [Phase 1]

Time Frame: Within first 4 weeks of treatment. DLT Criteria can include:

^■ > Gr 3 non-hematologic toxicity or laboratory finding

^■ Gr 4 hematologic toxicity by Day 42 in absence of disease

^■ Inability to tolerate at least 75% of Cycle 1 treatment

3. Doses recommended for future studies [Phase 1] Time Frame: Within first 4 weeks of treatment.

4. Complete Response (CR, CRi, MLFS, Marrow CR) Rate of Compound 1 as a single agent or in combination with Azacitidine in patients with AML/MDS [Phase 2]

Time Frame: As per IWG Response Assessment Guidelines for AML and MDS based on investigator's assessment through study completion, e.g. modified IWG AML 2003/MDS 2006.

Secondary Outcome Measures

[0265] The outcome of the study can also be evaluated using the following criteria: Area under the plasma concentration versus time curve (AUC) [Phase 1 and Phase 2] Time Frame: Blood samples for PK analysis collected at multiple visits during the first 60 days of treatment and on day 1 of all cycles following the first 30 days.

Peak Plasma Concentration (Cmax) [Phase 1 and Phase 2] Time Frame: Blood samples for PK analysis collected at multiple visits during the first 60 days of treatment and on day 1 of all cycles following the first 30 days.

Time of peak plasma concentration (Tmax) [Phase 1 and Phase 2] Time Frame: Blood samples for PK analysis collected at multiple visits during the first 60 days of treatment and on day 1 of all cycles following the first 30 days.

Time for half of the drug to be absent in blood stream following dose (T 1/2) [Phase 1 and Phase 2] Time Frame: Blood samples for PK analysis collected at multiple visits during the first 60 days of treatment and on day 1 of all cycles following the first 30 days. Rate at which drug is removed from blood stream (CL/F) [Phase 1 and Phase 2]

Time Frame: Blood samples for PK analysis collected at multiple visits during the first 60 days of treatment and on day 1 of all cycles following the first 30 days.

Rate of drug distribution within the blood stream (Vd/F) [Phase 1 and Phase 2]

Time Frame: Blood samples for PK analysis collected at multiple visits during the first 60 days of treatment and on day 1 of all cycles following the first 30 days.

Reduction of 2-HG levels in plasma [Phase 1 and Phase 2] Time Frame: Blood samples for PK/PD analysis collected at multiple visits during the first 60 days of treatment and on day 1 of all cycles following the first 30 days.

Evidence of antileukemic or antimyelodysplastic activity of Compound 1 as determined by complete response (CR), CRi (complete remission with incomplete hematologic recovery), morphologic leukemia-free state (MLFS), Marrow CR, partial remission (PR), and stable disease (SD) as a single-agent or in combination with azacitidine or cytarabine [Phase 1] Time Frame: As per IWG Response Assessment Guidelines for AML and MDS based on investigator's assessment through study completion.

Incidence and severity of adverse events, clinical laboratory abnormalities, and changes in ECG parameters as assessed by CTCAE v4.0 as a single-agent or in combination with azacitidine [Phase 2] Time Frame: Safety will be assessed from time of first dose through 28 days post last dose. 10. Additional measures of antileukemic or antimyelodysplastic activity as determined by CRh, Overall Response (OR), and Stable Disease of Compound 1 alone or in

combination with azacitidine [Phase 2] Time Frame: As per IWG Response Assessment Guidelines for AML and MDS based on investigator's assessment through study completion.

11. Time to Response (TTR) [Phase 2] Time Frame: From first dose of study drug through time of first response by blood recovery count.

12. Duration of Response (DOR) [Phase 2] Time Frame: From time of first response by blood recovery count through relapse.

13. Event-Free Survival (EFS) [Phase 2] Time Frame: From time of entry on study through progression.

14. Overall Survival (OS) [Phase 2] Time Frame: From time of entry on study through death or date last known alive at end of follow-up.

Disease History and Baseline Characteristics of Participants

[0266] A summary the disease history and participant demographics is provided below:

Table 16

* Including 3 pts treated with 100 mg QD with food.

**Not inclusive of all types; pt could have received more than one type

[0267] A summary of the baseline disease characteristics is shown below:

Table 17

*As reported by investigator per local tests **One pt with R100 mutation

[0268] A summary of the Investigator-Assessed Response is shown below:

Table 18

*one pt excluded from efficacy analysis due to the lack of a R132X mutation;

pt continued on treatment and achieved a marrow CR

** ORR = overall response rate (CR/CRm + CRh + Cri + MLFS + Marrow

CR)

[0269] Exemplary AML patient disposition is summarized in Table 19.

Table 19

aOther reasons include lack of response (for monotherapy and combination therapy groups); entering hospice, and alternative treatments (for combination therapy group)

[0270] Non-hematologic and hematologic TEAEs are summarized in Table 20 and Table 21 below. No DLTs were observed in dose escalation cohorts. IDH differentiation syndrome (IDH- DS) adverse events were experienced by 4 monotherapy patients (13%; grade 3,3; grade 2,1) and 6 combination therapy patients (13%; grade 3,3; grade 2,1). Most (7) were observed during cycle 1 (2 in cycle 2, 1 in cycle 5). All cases resolved with treatment interruption/reduction, dexamethasone, and/or supportive treatment. No recurrences were observed. Of the patients who experienced a IDH-DS AE, 2 of 4 monotherapy patients and 5 of 6 combination therapy patients achieved a response. QT prolongation was reported in 3 combination therapy patients (7%); all events were transient, and patients resumed treatment with no recurrences. Five of 32 patients in monotherapy and 5 of 46 patients in combination therapy had grade > 3 LFT (ALT, AST, TB) abnormalities, and two patients (1 monotherapy, 1 combination therapy) discontinued treatment due to these events.

Table 20

Table 21

[0271] Clinical response of patients with AML is summarized in Figure 9.

[0272] Survival of patients with AML is summarized in Figure 10.

[0273] 97% of patients had > 1 co-mutation, and 39% of patients had > 3 co-mutations

(Figure 11). [0274] Exemplary MDS patient disposition is summarized in Table 22. Median time on treatment for all patients with MDS treated with olutasidenib (± AZA) was 8.0 months.

Table 22

[0275] A summary of clinical activity in MDS patients is summarized in Table 23.

Table 23

a Efficacy evaluable population. One patient was excluded from efficacy analysis due to lack of

R132X mutation. ^b ORR = CR + marrow CR + PR. NR, not reached.

[0276] Figure 12 summarizes time on treatment for patients with MDS.

[0277] Table 24 summarizes hematologic improvement in clinical benefit and marrow CR for MDS patients. Table 24.

R132X mutation.

[0278] TEAEs (> 20% overall) in MDS patients are summarized in Table 25. IDH- differentiation syndrome was observed in 3 (13%) of patients. LFT (AST, AST, bilirubin) abnormalities were observed in 4 pts; 2 (Gl, G3) continued dosing through the elevation, 1 (G3) improved with dose reduction, and 1 (G3) discontinued treatment after a positive re-challenge.

Table 25

[0279] Twenty -two mutations were identified in 17 patients with samples available for central analysis (see Figure 13). Three patients had multiple baseline IDH1 mutations.

Dosing

[0280] The present disclosure includes, among other things, the novel understanding that administration of 300 mg of Compound 1 (e.g., 150 mg BID or 300 mg QD) to a patient or population of patients results in a sustained therapeutically effective trough blood plasma concentration (Css). Such a Css of Compound 1 resulted in a durable reduction in 2-HG plasma level over the course of at least 6 treatment cycles.

[0281] As outlined in Example 11, the concentration total plasma concentration of

Compound 1 and the plasma concentration of 2-HG was measured in the blood of patients receiving one of three different dose and dose intervals: 150 mg QD, 300 mg QD or 150 mg BID (either receiving Compound 1 as a single agent or in combination with azacitidine as described in the clinical trial of Example 11, in each category). The 2-HG levels were measured prior to administration of Compound 1, and then measured after administration of Compound 1 up to cycle 2, day 1 after first receiving Compound 1 (as the solid form obtained from Example 5).

[0282] As shown in Figure 14 and Figure 15, the administration of Compound 1 at 150 mg BID resulted in a trough blood plasma concentration above 1,652 ng/mL after cycle 2 of a 28- day treatment cycle as both a single agent and in combination with azacitidine. Additionally, as shown in Figure 16 and Figure 17, a reduction of 2-HG level was observed. The decrease in 2- HG concentration observed in patients receiving Compound 1 at 150 mg BID or 300 mg QD is unexpectedly better than those who received Compound 1 at 150 mg QD, both in terms of magnitude and variability of effect.

[0283] As shown in Figure 18A-18E, the plasma exposures (steady state blood plasma concentration) of Compound 1 were durable (i.e., sustained) throughout at least a 6 cycle treatment duration. As shown in Figures 19A-19D the plasma 2-HG concentrations were reduced within 1 cycle (C2D1) and maintained throughout the treatment duration. Variant Allele Frequency (VA l·) Analysis - AML

[0284] 229 samples (213 from white blood cells (PaxGene and EDTA) and 16 from bone marrow analysis) were obtained from 59 AML patients treated with either Compound 1 as a single agent or Compound 1 in combination with azacitidine in the Phase 1 study. Next generation sequencing was carried out through target enrichment using HaloPlex® Target followed by Illumina® sequencing; coverage > 100X across 75 genes. Digitial droplet PCR (ddPCR) was carried out through an input of 20 ng on a Stilla 3 -channel system; VAF data based on > 20,000 droplets.

[0285] As shown in Figure 25, good correlation between ddPCR and NGS was observed, which justifies using ddPCR for on-treatment assessment of IDH1 VAF. As shown in Figure 26, detection of IDH1 from BMA can be useful in patients with low IDH1 VAF in WB.

[0286] Of the 59 pts with local and central IDHlm results (all sample types included), 53/59 (90%) central testing confirmed presence of IDHlm at study entry.

[0287] As shown in Figure 27A, Figure 27B, Figure 28, and Figure 29, upon treatment with Compound 1, significant reduction in IDH1 VAF across categories was observed. 25 patients achieved an objective response and 6 patients with SD had available longitudinal samples for analysis (VAF at > C3). IDH1 mutation clearance / significant reduction is observed in 10/25 (40%) patients with an IWG response to Compound 1 (Figure 27A). In patients with stable disease, 3/6 (50%) had clearance / significant reduction of the IDHlm VAF (Figure 27B).

Figures 28 and 29 show that clinical response is associated with a decrease in 2-HG and clearance of the IDHlm clone.

VAF Analysis MDS

[0288] As shown in Figure 30, treatment with Compound 1 induces IDH1 mutation clearance and reduces 2-HG levels. 44% of patients (4/9) experienced mutation clearance (VAF <1%; only patients who received > 3 cycles of treatment and > 1 post-baseline VAF assessment were analyzed for mutation clearance). Rapid reduction of 2-HG was observed by end of cycle 1

Mechanism Resistance/Escape

[0289] The following details two case studies of individual patients.

Case Study 1: IDH2-Mediated Resistance [0290] As shown in Figure 31, an R/R AML patient with known IDH2m at baseline was treated with Compound 1 in combination with azacitidine. The patient remained in stable disease for 6 cycles then progressed. Compound 1 induced clearance of the IDHlm clone, however azacitidine was not effective in controlling the IDH2m clone that eventually drove the clinical progression.

Case Study 2: Presence of Additional Non-IDHlm Clones Drive Resistance

[0291] A treat naive AML secondary to MDS patient treated with Compound 1 as a single agent. As shown in Figure 32, this patient remained stable for 15 cycles with no achievement of an IWG response, however IDH1 mutation clearance and normalization of 2-HG were observed.

Conclusion

[0292] Compound 1 demonstrates clinical activity as a single agent and in combination with azacitidine in a high-risk Phase 1 population of AML/MDS patients with IDH1 mutation. In R/R AML, 41% and 46% pts achieve ORR with Compound 1 as a single agent and Compound 1 in combination with azacitidine treatment, respectively. 90% of pts enrolled with a history of IDHlm determined locally had a IDHlm confirmed centrally. Baseline co-mutation analyses demonstrated no correlation with clinical response (likely due to the small number of patients). Compound 1 induces IDH1 mutation clearance or significant reduction in treatment naive and R/R AML patients regardless of IWG response. Of the 25 patient that achieved an objective response and with available samples (VAF at > C3), 10 (40%) had clearance or significant VAF reduction to < 1%. Six stable disease patients had samples available and three (50%) had clearance or significant VAF reduction. Initial analysis of patients who relapse/progress on Compound 1 suggests non-IDHm-driven mechanism of escape.

[0293] Compound 1 is well-tolerated as a single agent and in combination with AZA in patients with MDS. Patients with MDS remained on treatment for a median of 8 months.

Compound 1 demonstrated preliminary clinical activity as a single agent and in combination with AZA in treatment naive and relap sed/refractory patients with MDS. 50% (3/6) patients treated with Compound 1 monotherapy achieved CR/mCR; 56% (9/16) patients treated with Compound 1 + AZA achieved CR/mCR. Clinical benefit with hematologic improvement was observed with Compound 1 monotherapy and combination therapy in 17% and 31% of patients, respectively. Mutation clearance was observed in 44% of evaluable patients. Rapid and sustained reduction of 2-HG was seen by the end of cycle 1. Example 12: A Phase lb/2 Study of Compound 1 in Patients with Advanced CNS and Solid Tumors with an IDH1 Mutation.

[0294] Patients having any of the following solid tumors that harbor a IDHl mutation received Compound 1 (unless otherwise indicated, at a dose of 150 mg of the solid form provided in Example 5, administered orally BID) as a single agent or in combination with additional therapies:

• Compound 1 was administered to patients diagnosed with glioma or chondrosarcoma as a single agent or in combination with 5-azacitidine (also referred to herein as“5- azacytidine” or“azacytidine” or“azacitidine”);

• Compound 1 was administered to patients diagnosed with hepatobiliary cancers as a single agent or in combination with a PD-1 inhibitor (preferably, nivolumab);

• Compound 1 was administered to patients diagnosed with intrahepatic

cholangiocarcinoma as a single agent or in combination with gemcitabine and cisplatin (“GemCis”); and

• Compound 1 was administered to patients diagnosed with other non-CNS solid tumors as a single agent.

[0295] Each patient had a histologically or cytologically-confirmed IDHl R132X gene- mutated advanced solid tumor prior to receiving Compound 1. In particular, some patients receiving Compound 1 had a histologically or cytologically-confirmed IDHl R132X gene- mutated advanced glioma that has recurred or progressed following standard therapy. Patients receiving Compound 1 can have relapsed or refractory glioma (per WHO criteria 2016) with confirmed IDHl mutation. Other patients receiving Compound 1 had relapsed or refractory hepatobiliary tumors with confirmed IDHl mutation previously treated with an approved therapy for HBC. Other patients receiving Compound 1 had recurrent, refractory or either locally advanced or metastatic chondrosarcoma with confirmed IDHl mutation not amenable to complete surgical excision. Other patients receiving Compound 1 had advanced, nonresectable or metastatic intrahepatic cholangiocarcinoma with confirmed IDHl mutation not eligible for curative resection or transplantation. Some patients receiving Compound 1 had relapsed or refractory other solid tumors with confirmed IDHl mutation.

[0296] Patients were assessed for pharmacokinetics (PK) (e.g., by collecting a blood sample) at regular intervals throughout a course of treatment. In particular, pre-dose PK assessment was performed at least on days 1, 2, 8, 15, and 22 of the course of treatment for patients having a course of treatment comprising one or more 28-day treatment cycles. (Additional post dose assessment can be performed at cycle 1 day 1 and cycle 2 day 1.) In addition, pre-dose PK assessments were collected on day 1, 2 and 15 of cycle 2 of a 28-day treatment cycle during the course of treatment. Additional pre-dose PK assessment was performed on day 1 of Cycle 3 and subsequent 28-day treatment cycles during the course of treatment.

[0297] Compound 1 was administered to patients who met the following criteria for inclusion: > 18 years of age; Life expectancy of > 4 months; Documented IDH1 gene-mutated malignancy based on local test evaluation; Able to provide tumor tissue sample (archival); and cancer diagnosis as described above. Preferably, patients also met one or more of the following additional inclusion criteria:

• Recovered to < Grade 2 or baseline toxicity (except alopecia) from prior therapy (per CTCAE v 4.03);

• Eastern Cooperative Oncology Group (ECOG) performance status 0-2;

• Adequate bone marrow function (e.g., Absolute neutrophil count (ANC) > 1.5 x 109/L without any growth factors in prior 7 days and Hemoglobin > 8.0 g/dL (with or without transfusion support); Platelet count > 75 x 10⁹/L (with or without transfusion support); Cohort 4b (GemCis combination); and platelet count > 100 x 109/L (with or without transfusion support));

• Child-Pugh Class A (HBC only);

• Adequate hepatic function (e.g, Aspartate aminotransferase (AST) (serum glutamic oxaloacetic transaminase [SGOT]) / alanine aminotransferase (ALT) (serum glutamic pyruvate transaminase [SGPT]) < 2.5 x institutional upper limit of normal (ULN). For patients with suspected malignancy related elevations, < 5 x ULN; and Total bilirubin < 1.5 x ULN. For patients with suspected malignancy related elevation < 3 x institutional upper limit of normal); and

• Adequate renal function (e.g, Creatinine clearance per Cockcroft-Gault equation of > 60 mL/min).

[0298] Compound 1 was not administered to patients who met one or more of the following exclusion criteria:

• Previous solid organ or hematopoietic cell transplant • Prior anticancer treatment (e.g, No prior treatment with small molecule, antibody, or other anticancer therapeutic within 21 days (or 5 half-lives), whichever is shorter of first dose of study treatment (6 weeks for nitrosoureas or mitomycin C); No previous treatment with an IDH1 inhibitor; No prior radiation therapy (including radiofrequency ablation) within 4 weeks prior to initiation of study treatment; No prior stereotactic body radiation therapy (SBRT) within 2 weeks prior to initiation of study treatment; and No prior chemoembolization or radioembolization within 4 weeks)

• Congestive heart failure (New York Heart Association Class III or IV) or unstable angina pectoris; previous history of myocardial infarction within one year prior to study entry, uncontrolled hypertension, or uncontrolled arrhythmias

• History of QT prolongation or baseline QT interval corrected with Fridericia’s method (QTcF) > 450 ms (average of triplicate readings) (NOTE: criterion does not apply to patients with a right or left bundle branch block)

• Concomitant medication(s) associated with QTc interval prolongation or Torsades de Pointes (TdP) initiated less than the duration required to reach steady-state plasma concentration (approximately five half-lives) before first dose of study drug (medications used as needed [PRN] (e.g. Zofran) are exempt).

• Pregnant or nursing women or women of childbearing potential not using adequate

contraception; male patients not using adequate contraception

• Other malignancy within the last 5 years except: adequately treated non-melanoma skin cancer, curatively treated in situ cancer of the cervix, ductal carcinoma in situ (DCIS), stage 1, grade 1 endometrial carcinoma, or other solid tumors including lymphomas (without bone marrow involvement) curatively treated with no evidence of disease for >

5 years

• Major surgery within 4 weeks of starting study treatment or not recovered from any

effects of prior major surgery (uncomplicated central line placement or fine needle aspirate are not considered major surgery);

• Patients receiving > 6 mg/day of dexamethasone or equivalent

• Patients with gastrointestinal disorders likely to interfere with absorption of the study medication

• Known history of HIV positivity • Active infection with hepatitis B or C virus (Hep B or C viral load>100 international units/milliliter or local institutional equivalent)

• Unstable or severe uncontrolled medical condition (e.g., unstable cardiac function,

unstable pulmonary condition including pneumonitis and/or interstitial lung disease, uncontrolled diabetes) or any important medical or psychiatric illness or abnormal laboratory finding that would, in the Investigator’s judgment, increase the risk to the patient associated with his or her participation in the study and

• PD-1 combination only: Patients with active autoimmune disease (Note: patients with well controlled diabetes or hypothyroidism are eligible).

[0299] Each patient received Compound 1 throughout a medically appropriate course of treatment. In general, patients received Compound 1 (either single-agent Compound 1 or combination therapy as indicated above) until disease progression or unacceptable toxicity. At the start of the course of treatment, each patient received Compound 1 as the solid form obtainable by the method of Example 5 at a dose of 150 mg BID administered continuously in one or more 28-day treatment cycles.

[0300] The DLT-Evaluable Analysis Set was defined as all patients in the Safety Lead-in Periods (single agent Compound 1, combination Compound 1 + 5-azacitidine, combination Compound 1 + GemCis and combination Compound 1 + PD-1 inhibitor such as nivolumab) who either experienced a DLT during Cycle 1 or completed at least 75% of the prescribed Cycle 1 dose. These analysis sets were used to assess the tolerability of Compound 1.

[0301] The Safety Analysis Set was defined as all patients who received any amount of study drug(s) (Compound 1 and combination agents, if appropriate). This analysis set was the primary analysis set for all safety endpoints, excluding DLT evaluation. Safety analysis was by cohort and by treatment within cohort if more than one dose or dosing combination were initiated for a particular indication cohort.

[0302] The Response-Evaluable Analysis Set was defined as all patients with measurable disease at baseline who received the study drug(s) specific to the part of their particular cohort (either Compound 1 monotherapy or Compound 1 in combination), and had at least 1 post baseline response assessment or discontinued the treatment phase due to disease progression (including death caused by disease progression) within 8 weeks (+2 -week window) of the first dose of study treatment. This analysis set was the primary analysis set for efficacy endpoints such as ORR. All response evaluations were by cohort, and by treatment within cohort if more than 1 doses or dosing combinations are initiated for a particular indication cohort.

[0303] Patient safety measurements and clinical laboratory measurements were performed throughout the course of treatment for each cohort. Safety measurements included assessment of patient concomitant medications and procedures, AE/SAE assessment, symptom-directed physical examination and ECOG performance status. Clinical laboratory measurement assessment included blood chemistry and hematology and other measurements specific to individual cohorts.

[0304] For patients receiving Compound 1 as a single agent, or in combination with 5- azacitidine (glioma, chondrosarcoma) or in combination with a PD-1 inhibitor (HBC),

Compound 1 was administered in a 28-day treatment cycle (28 consecutive days, at a dose of 150 mg BID) and patient safety measurement and clinical blood chemistry and hematology were obtained on the following days during the course of treatment for patients in the first 28-day treatment cycle during the clinical trial: day 1, day 8 (+/- 2), day 15 (+/- 2), day 22 (+/- 2). In treatment cycle 2 and beyond, these assessments were obtained at day 1 {+1-2) and day 15 {+1-2).

[0305] For patients receiving Compound 1 in combination with chemotherapy (e.g., GemCis for cholangiocarcinoma), Compound 1 was administered in a 28-day treatment cycle (28 consecutive days, at a dose of 150 mg BID) and patient safety measurement and clinical blood chemistry and hematology were obtained on the following days during the course of treatment for patients in the first six 28-day treatment cycle during the clinical trial: day 1, day 8 {+1-2), day 15 {+1-2), day 22 {+1-2). The combination was given for a total of six treatment cycles. In treatment cycle 7 and beyond, these assessments are obtained at day 1 {+1-2) and day 15 {+1-2). The patient could continue on single agent Compound 1 treatment without combination agent as directed by treating physician.

Safety Lead-In Period

[0306] The study included a Safety Lead-in Period where single-agent 150 mg Compound 1 BID was administered over 28 days (1 cycle). The Safety-Lead-in Period employed a traditional 3+3 design, whereby 3 patients with solid tumors and 3 patients with gliomas were treated with Compound 1 150 mg BID and monitored for dose-limiting toxicities (DLTs) during the first cycle of study treatment. • If no DLTs occur in the first 3 patients in either group (solid tumors or glioma), and available pharmacokinetic (PK)/pharmacodynamic (PD) data support the dose, enrollment continues in the 4 disease-specific cohorts described below.

• If a DLT occurs in the first 3 patients in either group, an additional 3 patients are treated at that dose level in the relevant group. If no DLTs occur in these additional 3 patients (i.e.< 2 DLTs per 6 patients) and available PK/PD data support the dose, enrollment continues in the 4 disease-specific cohorts described below.

• If there are > 2 DLTs at the starting dose, lower doses or an altered dosing schedule of Compound 1 can be considered. Likewise, higher doses may be evaluated based upon safety, PK, and PD data as determined by the SRC.

Cohort 1: Glioma (n = 16-31)

[0307] Compound 1 is used to treat patients diagnosed with a glioma cancer diagnosis. In particular, Compound 1 is administered to patients meeting the inclusion criteria above and one or more the following disease related inclusion criteria: histologically or cytologically confirmed IDH1 gene-mutated advanced glioma, and a diagnosis of glioblastoma multiforme with confirmed IDHl gene-mutated disease with first or second recurrence. Cohort 1 includes patients with glioma harboring an IDHl mutation that is relapsed or refractory. Glioma patients are treated with single-agent Compound 1 (Cohort la). Cohort la employs a Simon’s 2-stage design, in which 8 patients are treated with single-agent Compound 1 for a minimum of 4 cycles (cycle = 28 days) and assessed for efficacy and safety (Stage 1). If > 1 clinical response is observed in Stage 1, then Stage 2 (n=15) initiates with single-agent Compound 1. If no clinical responses are observed in Stage 1 with single-agent Compound 1, then combination therapy is examined (Compound 1+ 5-azacytidine) (Cohort lb). Cohort lb employs a Simon’s 2-stage design, whereby 8 patients are treated in Stage 1 with combination therapy for a minimum of 4 cycles (cycle = 28 days) and assessed for efficacy and safety. If > 1 clinical response is observed in Stage 1 of Cohort lb, then Stage 2 (n=15) is initiated with combination therapy. During Stage 1 aggregate safety data are monitored by the SRC. If unacceptable toxicity is observed in Stage 1, then the dose and schedule may be modified by the SRC. (Note: any glioma patients enrolled in the safety Lead-in Period are considered part of Stage 1 enrollment.) Cohort 2: Hepatobiliary Carcinoma (HBC) ( = 21-63 )

[0308] Compound 1 is used to treat patients diagnosed with a hepatobiliary carcinoma (HBC) cancer diagnosis. In particular, Compound 1 is administered to patients meeting the inclusion criteria above and one or more the following disease related inclusion criteria:

Relapsed/refractory or intolerant to approved standard-of-care therapy (included: hepatocellular carcinoma, biliary carcinoma or other hepatobiliary carcinomas); Histologically or cytologically confirmed IDHl gene-mutated with measurable disease per RECIST 1.1 criteria; and Child-Pugh Class A.

[0309] Cohort 2 includes patients with relap sed/refractory HBC harboring an IDHl mutation. HBC patients are initially treated with single-agent Compound 1 (Cohort 2a). Prior exposure to nivolumab is permitted for patients of Cohort 2a. Cohort 2a employs a Simon’s 2- stage design, in which 8 patients are treated with single-agent Compound 1 for a minimum of 4 cycles (cycle = 28 days) and assessed for efficacy and safety (Stage 1). If > 1 clinical response is observed in Stage 1, then Stage 2 (n=15) is initiated with single-agent Compound 1. If no clinical responses are observed in Stage 1 with single-agent Compound 1, then combination therapy can be examined (Compound 1 + PD1 inhibitor) (Cohort 2b). Cohort 2b employs a Simon’s 2-stage design, whereby 13 patients are treated in Stage 1 with combination therapy for a minimum of 4 cycles (cycle = 28 days) and assessed for efficacy and safety. If > 4 clinical response is observed in Stage 1 of Cohort 2b, then Stage 2 (n=42) can initiate with combination therapy. Prior exposure to nivolumab is not permitted for patients of Cohort 2b. During Stage 1 aggregate safety data is monitored by the SRC. If unacceptable toxicity is observed in Stage 1, then the dose and schedule may be modified by the SRC. (Note: any HBC patients enrolled in the Safety Lead-in Period are considered part of Stage 1 enrollment.)

Cohort 3: Chondrosarcoma ( h = 16-31)

[0310] Compound 1 is used to treat patients diagnosed with a chondrosarcoma cancer diagnosis. In particular, Compound 1 is administered to patients meeting the inclusion criteria above and one or more the following disease related inclusion criteria: Relapsed or refractory and either locally advanced or metastatic and not amenable to complete surgical excision; and histologically or cytologically confirmed IDHl gene-mutated with measurable disease per RECIST 1.1 criteria. Cohort 3 includes patients with relapsed/refractory, locally advanced or metastatic chondrosarcoma harboring an IDHl mutation. Chondrosarcoma patients are treated with single-agent Compound 1 (Cohort 3a). Cohort 3a will employ a Simon’s 2-stage design, in which 8 patients will be treated with single-agent Compound 1 for a minimum of 4 cycles (cycle = 28 days) and assessed for efficacy and safety (Stage 1). If > 1 clinical response is observed in Stage 1, then Stage 2 (n=15) will initiate with single-agent Compound 1. If no clinical responses are observed in Stage 1 with single-agent Compound 1, then combination therapy is examined (Compound 1 + 5-azacytidine) (Cohort 3b). Cohort 3b employs a Simon’s 2-stage design, whereby 8 patients are treated in Stage 1 with combination therapy for a minimum of 4 cycles (cycle = 28 days) and assessed for efficacy and safety. If > 1 clinical response is observed in Stage 1 of Cohort 3b, then Stage 2 (n=15) initiates with combination therapy. During Stage 1 aggregate safety data are monitored by the SRC. If unacceptable toxicity is observed in Stage 1, then the dose and schedule may be modified by the SRC. (Note: any chondrosarcoma patients enrolled in the Safety Lead-in Period are considered part of Stage 1 enrollment.)

Cohort 4: Intrahepatic Cholangiocarcinoma ( n = 21-63 )

[0311] Compound 1 is used to treat patients diagnosed with an intrahepatic

cholangiocarcinoma (IHCC) cancer diagnosis. In particular, Compound 1 is administered to patients meeting the inclusion criteria above and one or more of the following disease related inclusion criteria: Advanced nonresectable or metastatic IHCC not eligible for curative resection or transplantation; Phase lb/Safety Lead-in of Phase 2: relapsed or refractory disease; and histologically or cytologically confirmed IDH1 gene-mutated with measurable disease per RECIST 1.1 criteria. Cohort 4 includes patients with advanced IHCC harboring an IDH1 mutation. IHCC patients are treated with single-agent Compound 1 (Cohort 4a). Patients of cohort 4a must be ineligible for standard therapies. Cohort 4a employs a Simon’s 2-stage design, in which 8 patients are treated with single-agent Compound 1 for a minimum of 4 cycles (cycle = 28 days) and assessed for efficacy and safety (Stage 1). If > 2 clinical responses are observed in Stage 1, then Stage 2 (n=15) initiates with single-agent Compound 1. If < 2 clinical responses are observed in Stage 1 with single-agent Compound 1, then combination therapy is examined (Compound 1 + GemCis) (Cohort 4b). Patients in Cohort 4b have received no more than one cycle of Gem/Cis therapy. Cohort 4b employs a Simon’s 2-stage design, whereby 13 patients are treated in Stage 1 with combination therapy for a minimum of 4 cycles (cycle = 21 days) and assessed for efficacy and safety. If > 4 clinical responses are observed in Stage 1 of Cohort 4b, then Stage 2 (n=42) initiates with combination therapy. During Stage 1 aggregate safety data are monitored by the SRC. If unacceptable toxicity is observed in Stage 1, then the dose and schedule may be modified by the SRC. (Note: any IHCC patients enrolled in the Safety Lead-in Period are considered part of Stage 1 enrollment.)

[0312] In some examples, patients diagnosed with relap sed/refractory IHCC receive Compound 1 single agent, whereas patients newly diagnosed and treatment naive IHCC receive Compound 1 in combination with GemCis chemotherapy.

Cohort 5: Other non-CNS solid tumors with IDH1 mutations (n=12)

[0313] Compound 1 is used to treat patients diagnosed with non-CNS solid tumors with IDH1 mutations (preferably detectable 2-HG in plasma). Cohort 5 includes patients with relapsed or refractory non-CNS solid tumors harboring an IDH1 mutation. In particular, Compound 1 is administered to patients meeting the inclusion criteria above and one or more of the following disease related inclusion criteria: relapsed or refractory to standard-of-care therapy with no other available therapeutic options; and histologically or cytologically confirmed IDH1 gene-mutated with measurable disease per disease appropriate response criteria. This cohort includes treatment with single agent Compound 1. Due to the diverse population, this is an exploratory cohort without pre-defmed efficacy/futility determinations. Aggregate safety data is monitored by the SRC and if unacceptable toxicity is observed in > 2 patients, the cohort can be closed for additional enrollment.

[0314] For Cohorts 1-5 above, the efficacy assessments obtained include those listed in the table below. (MRS = 1H magnetic resonance spectroscopy; MRI = magnetic resonance imaging).

Table 26

Results - Glioma

[0315] Isocitrate dehydrogenase 1 mutations (mlDHl) are present in > 70% of patients with Grade II/III gliomas resulting in production and accumulation of (R)-2-hydroxyglutarate causing DNA hypermethylation and promoting tumorigenesis. Patients (pts) with relapsed/refractory (R/R) mlDHl gliomas received Compound 1 150 mg BID, orally either as single agent (SA) or in combination (CO) with azacitidine in a dose confirmation Phase lb followed by efficacy evaluation Phase 2 study (NCT: 03684811).

[0316] As of 12-Sept-2019, 24 pts with glioma (grade at enrollment: II/III/IV; n =4, n=12, and n=8, respectively) were treated with Compound 1 monotherapy. The median age was 46 years (range: 23-64); 63% were men and 38% were women. Patients had a median of 2 prior treatments, and 88% had received prior temozolomide. IDH1 mutation status was locally determined (IHC, NGS, or PCR). Baseline patient characteristics are summarized below. Values are presented as n (%) except for age and prior regimens.

Table 27

[0317] Treatment-emergent adverse events (TEAEs) are summarized below. No DLTs were observed in dose-confirmation Phase lb study with single agent Compound 1. Transaminase elevations resolved without sequelae in all patients. No QTc prolongation was reported as an adverse event. Twelve patients discontinued - 11 for disease progression and 1 for grade 4 acute hepatitis.

Table 28

a All Grade 3.

[0318] Duration of Compound 1 monotherapy treatment in predominantly enhancing gliomas is shown in Figure 33. Median duration of treatment at time of data cut was 3.7 months (range: 0.9-9.4).

[0319] Best percent change in tumor burden with Compound 1 monotherapy per investigator assessment (RANO) is shown in Figure 34.

[0320] Percent change in tumor burden with Compound 1 monotherapy per central volumetric assessment (BCIR) is shown in Figure 35.

[0321] Best overall response for patients treated with Compound 1 monotherapy is summarized in Table 29.

Table 29

a _I One enrolled patient did not have measurable disease and was not included in efficacy analysis.

[0322] Figure 36 summarizes local biomarker testing and best clinical response. Most tumors had >1 co-mutation as determined from tumor biopsy. 11/24 patients had no mutation testing done.

[0323] Compound 1 plasma concentrations in glioma patients (n=24) reached steady-state within 2 weeks of initiation of dosing and remained consistent over time. At steady-state, Compound 1 concentrations were below those predicted to pose a QT prolongation risk and above the concentration predicted to reduce 2-HG levels by 90% (based on preclinical data described above). Results are summarized in Figure 37.

[0324] Plasma D-2HG was measured at baseline and during the course of treatment.

Baseline and maximum reduction (after a minimum of 28 days of Compound 1 treatment) are shown for 15 disease-evaluable patients with paired samples. Results are summarized in Figure 38A and Figure 38B. Median duration of treatment at time of data cut was 3.7 months (range: 0.9-9.4). [0325] Compound 1 was measured in patient plasma and CSF. Results are summarized in Table 30. Compound 1 human plasma protein binding was 94.5% (5.5% unbound), and the unbound brain partition coefficient (Kpuu) for Compound 1 in humans was 0.54. This confirms that Compound 1 is brain penetrant when administered at a clinically relevant dose. Compound 1 CSF concentration is predicted to be at the IC75 for 2-HG reduction.

Table 30

[0326] Compound 1 at 150 mg BID demonstrates acceptable safety profile in patients with relapsed/refractory IDH1 -mutated glioma. Steady-state Compound 1 plasma concentrations were above preclinical minimum effective concentration and below concentrations predicted to pose a QT prolongation risk in non-human primates. CNS penetration was demonstrated by CSF exposures and disease control, thus confirming preclinical experiments (e.g., as described herein). Nine of 15 patients with glioma demonstrated a reduction of 2-HG at steady-state in plasma; however, 2-HG concentrations in plasma may not provide an accurate estimation of 2- HG reduction in tumor. Efficacy results suggest that Compound 1 monotherapy shows positive clinical activity and disease control in a population with enhanced relap sed/refractory IDHl- mutated glioma.

[0327] As of 31 -Oct-2019, 29 pts with R/R mIDHI glioma were treated with Compound 1 as SA (n = 24) or CO (n = 5). The median age was 45 yrs (range: 23-64) & 62% were male. WHO Glioma Grade (Gr) at study entry was: II (17%), III (52%) & IV (31%). Median number of prior treatments was 2 (1-5); 86% had received prior temozolomide. mIDHI status was locally determined (IHC, NGS or PCR): R132H (86%), R132L (7%), R132C (3.5%) & unspecified (3.5%). The median duration of Compound 1 treatment for SA & CO was 4.8 (1-11.4) & 1 (0.2- 2.3) months, respectively. Fifteen pts discontinued (disease progression [n = 12], AE [n = 1], withdrew consent [n = 1], other [n = 1]). For SA, the most common (> 25%) TEAEs (all grades, regardless of attribution) were: fatigue (50%), nausea (50%), diarrhea (33%), ALT increase (29%) & headache (29%). For CO, TEAEs that occurred in > 2 pts were: nausea (n = 4), fatigue (n = 2), neutropenia (n = 2), ALT increase (n = 2) & AST increase (n = 2). There were 2 protocol defined DLTs in the CO cohort, 1 pt with Gr 4 ALT, Gr 3 AST & Gr 3 GGT elevations & 1 pt with Gr 3 ALT elevation. No pts experienced a TEAE of QTcF prolongation. SA best responses are shown in Table 31; CO pts are too early for response assessment. The median PFS for SA was 8.3 months. Twenty (87%) and 11 (48%) pts were alive and progression-free at 6 & 12 months, respectively.

[0328] SA Compound 1 at 150 mg BID demonstrates acceptable safety and tolerability with preliminary clinical activity in glioma pts. Evaluation of CO is ongoing.

Table 31

* 1 pt not evaluable

[0329] As of 2020-03-31, 44 patients with R/R mIDHI glioma were treated with Compound 1. Baseline demographics and disease characteristics are summarized in Table 32.

Table 32

[0330] Patient dispositions are summarized in Table 33.

Table 33

a Patient continuing on monotherapy at data cutoff.

[0331] Figure 39 shows Compound 1 plasma concentration levels. Consistent plasma concentrations at levels predicted to safely provide benefit were achieved and maintained over treatment duration. Co-administration with AZA did not alter pharmacokinetics of Compound 1.

[0332] Table 34 summarizes TEAEs (> 15%) reported. (TEAE cutoff applied to

monotherapy population, N = 26.) 6 (23%) monotherapy and 3 (50%) combination patients experienced > Grade 3 LFT abnormality. In two patients, this led to treatment discontinuation (1 monotherapy; 1 combination). Two DLTs (> Grade 3 transaminase elevations) were observed for combination treatment, which met stopping criteria. Table 34

[0333] Figure 40 summarizes duration on treatment with Compound 1 monotherapy.

Disease control (PR + SD) was observed in 50% of patients, and 9 (38%) patients were stable for greater than 4 months. Median duration of treatment was 4.2 months (range: 0.03 to 15.9). In enhancing patients, median duration of treatment was 3.7 months (range: 0.03 to 15.9). Median time to first response was 1.9 months. Median duration of response was 10.1 months.

[0334] Investigator-assessed best overall response per RANO is summarized in Table 35 A.

Figure 41 shows % change from baseline in investigator assessment per RANO.

Table 35A

a Response-evaluable analysis set; 2 patients non-evaluable.

[0335] Independent central volumetric assessment of tumor burden (BICR) is summarized in Table 35B. Figure 42 shows % change from baseline for central volumetric assessment (BICR). Table 35B

a Response-evaluable analysis set; 2 patients non-evaluable; 2 patients had no data at data cutoff.

[0336] A case study of a 50 year old female patient diagnosed with mIDH (R132H) glioblastoma follows. The patient was diagnosed in 2014 and underwent treatment, including gross total resection, chemoradiation and temozolomide. The patient enrolled in a Compound 1 clinical trial in March 2019, where the patient was treated with Compound 1 150 mg BID. After 15 cycles, the patient remains on treatment, off steroids, and is clinically stable. Figure 43 shows a timeline of the patient’s therapy, including exemplary brain scans throughout the patient’s treatment with Compound 1.

[0337] Compound 1 was well tolerated as monotherapy in glioma patients. Compound 1 plasma concentrations were maintained over the treatment duration at levels predicted to safely provide benefit. Clinically relevant concentrations of Compound 1 were observed in the CSF, confirming CNS penetration observed in preclinical models. Compound 1 demonstrated preliminary disease control rate of 50% in a heavily pretreated patients with predominantly enhancing recurrent mlDHl glioma. Nine patients had SD for greater than 4 months. One patient achieved a PR per investigator assessment by RANO. Four patients achieved > 50% tumor reduction by blinded independent central volumetric assessment (BICR).

Results mlDHl Solid Tumors

[0338] Isocitrate dehydrogenase 1 mutations (mlDHl) are present in a variety of solid tumors resulting in production and accumulation of (R)-2- hydroxyglutarate causing DNA hypermethylation and promoting tumorigenesis. Patients with advanced relapsed/refractory (R/R) mlDHl solid tumors received Compound 1 150 mg BID orally. Following a dose confirmation cohort (Phase lb), patients with intrahepatic cholangiocarcinoma (IHCC), chondrosarcoma (CS), or unspecified mlDHl solid tumors (Other) were enrolled in a Phase 2 efficacy evaluation (NCT: 03684811). [0339] As of 31 -Oct-2019, 44 patients with relapsed or refractory mIDHI solid tumors were treated with Compound 1. Diagnosis included: IHCC (n = 26), CS (n = 13), and Other (n = 5). The median age was 58 years (range: 29-81) and 43% were male. Median number of prior treatments was 2 (1-10). mIDHI status was locally determined (IHC, NGS or PCR): R132C (61%), R132G (7%), R132S (7%), R132H (2%), R132L (2%), Others (2%) & unspecified (18%). Fourteen patients discontinued treatment (disease progression [n = 6; 3 IHCC, 2 CS, 1 Other], AE [n = 4; 3 IHCC, 1 CS], PI decision [n = 3; IHCC] & withdraw consent [n = 1;

IHCC]). Treatment emergent adverse events (all grades, regardless of attribution) that occurred in > 15% of pts were: nausea (43%), fatigue (25%), decreased appetite (22%), AST increase (18%), ALT increase (16%), and constipation (16%). No protocol-defined DLTs occurred. Best responses by tumor type are shown in Table 36.

[0340] Single agent Compound 1 at 150 mg BID demonstrates acceptable safety and tolerability with preliminary clinical activity in patients with R/R mIDHI solid tumors.

Table 36

* salivary tumor, lung adenocarcinoma, duodenal tumor (n = 1 for each) and cancer of unknown primary (n = 2)

Example 13: Case Studies of Human Clinical Trial Testing of Compound 1

[0341] Patient X is 66 y/o female, diagnosed with AML who initially received induction treatment with high dose cytarabine to which the patient was refractory. Subsequently, the patient enrolled in a clinical trial study, where she was treated with single agent (SA) Compound 1 150mg BID and achieved a complete remission (CR) after one cycle of treatment (28 days). Patient continued treatment while in CR for 7 additional cycles. Patient then relapsed and discontinued study treatment.

[0342] Patient Y is 62 y/o male, diagnosed with FLT3-positive secondary AML (secondary to MDS). Patient received intensive chemotherapy induction with cytarabine and daunorubicin in combination with midostaurin (FLT3 inhibitor) but unfortunately was refractory. He enrolled in a clinical trial study, where he was treated with Compound 1 150 mg BID in combination with azacitidine for a total of 8 cycles (1 cycle = 28 days). He achieved complete remission with IDH1 mutation clearance (CRm) by cycle 6 and discontinued study treatment after cycle 8 to undergo bone marrow transplant (HSCT).

[0343] Patient Z is a 50 year old diagnosed with grade III IDHlm glioma (anaplastic astrocytoma) previously treated with chemotherapy and radiation according to the applicable standard of care. This patient was subsequently enrolled on the clinical study treated with Compound 1 at 150 mg twice daily (BID) each day. Following treatment with Compound 1 for 2 cycles (each cycle=28 consecutive days receiving 150 mg Compound 1 BID), by MRI, patient was determined by the investigator to have experienced a partial response by RANO criteria (>50% decrease in tumor, no new lesions, on stable dose corticosteroids, no progression of measurable disease). After receiving 2 cycles of Compound 1 (150 mg BID), the patient remains on treatment with 150 mg BID Compound 1 per protocol.

[0344] 3 Patients received a Compound 1 at 100 mg once daily (QD) each day. Blood samples were collected every 28 days for measurement of plasma concentrations of Compound (single agent) Blood was collected at the following times relative to Compound 1 administration:

• Cycle 1 Day 1 : predose, and postdose at 30 minutes, 1, 2, 4, and 8 hours

• Cycle 1 Days 2, 8, 15, and 22: predose

• Cycle 2 Day 1 : predose, and postdose at 30 minutes, 1, 2, 4, and 8 hours

• Cycle 2 Day 2: 24 hours after C2D1 dosing (± 2 hours) for patients in dose expansion

• Cycle 2 Day 4: predose [72 hours after C2D1 dosing (± 4 hours)] for patients in dose expansion only

• Cycle 2 Day 15: predose

• Cycle 3 and Beyond: Predose on Day 1 of every cycle

[0345] The obseved Cmin associated with this case study can be found in Figure 14.

Example 14: Diagnostic for Identifying AML Patients Having a Susceptible mIDHI

Mutation

[0346] Abbott REALTIME IDHl is a commercially available, FDA-Approved in vitro polymerase chain reaction (PCR) assay for the qualitative detection of single nucleotide variants (SNVs) coding five IDHl R132 mutations (R132C, R132H, R132G, R132S, and R132L) in DNA extracted from human blood (EDTA) or bone marrow (EDTA). PMA Applicant: Abbott Molecular Inc., 1300 E. Touhy Avenue, Des Plaines, IL 60018; FDA Approval Date: July 20, 2018). Abbott RealTime IDHl is for use with the Abbott m2000rt System. It will be appreciated that further details on using IDHl Assay of this Example are available in product literature and instruction manuals accompanying the assay and/or the real-time PCR system.

[0347] The Abbott RealTime IDHl is indicated as an aid in identifying acute myeloid leukemia (AML) patients with an isocitrate dehydrogenase- 1 (IDHl) mutation for treatment with an FDA-Approved mIDHI inhibitor. This test is for prescription use only. The Abbott RealTime IDHl detects single nucleotide variants (SNVs) coding five IDHl mutations (R132C, R132H, R132G, R132S, and R132L) by using PCR technology with homogeneous real-time fluorescence detection. The assay uses human blood or bone marrow aspirate specimens and reports a qualitative result. The table below lists the IDHl mutations detected by the Abbott RealTime IDHl assay.

Table 37

Biological Principles of the Procedure

[0348] The IDHl Assay of this Example consists of two kits:

• IDHl amplification reagent kit

• IDHl control kit

[0349] Specimens for the IDHl Assay of this Example are processed manually using reagents (e.g., lysis buffer containing guanidine isothiocyanate, magnetic microparticles, wash buffers, and/or elution buffer) to isolate and purify sample DNA. The amplification reagents are combined into two amplification master mixes. The purified DNA sample is combined with the master mixes in a 96-well optical reaction plate, and the plate is transferred to a real-time PCR instrument for amplification and detection of IDHl mutations. The specimen result is automatically reported on a real-time PCR workstation at run completion. Assay controls are included within each run and are processed through DNA extraction, amplification, and detection steps of the assay to assess run validity.

[0350] Software parameters specific to the IDH1 Assay of this Example are contained in an assay application specification file, which is loaded onto a real-time PCR instrument by using a CD-ROM disk.

DNA Extraction

[0351] The purpose of DNA extraction is to isolate and purify genomic DNA from EDTA preserved blood or bone marrow aspirate specimens to make it accessible for amplification and to remove potential inhibitors of amplification. This process is accomplished by using magnetic particle technology to isolate and purify DNA. During the DNA extraction procedure, cells are lysed at an elevated temperature in a lysis buffer containing guanidine isothiocyanate. DNA is captured on magnetic microparticles, and inhibitors are removed by performing a series of washes with wash buffers. The bound DNA is eluted from the microparticles with elution buffer and is ready for PCR amplification.

Reagent Preparation and Reaction Plate Assembly

[0352] IDH1 oligonucleotide reagents (Oligonucleotide Reagent 1 and Oligonucleotide Reagent 2) are each manually combined with a DNA polymerase and activation reagent to create 2 unique master mixes. These master mixes are added to 2 separate wells of a 96-well optical reaction plate along with aliquots of the extracted DNA sample. After manual application of an optical adhesive cover, the plate is transferred to a real-time PCR instrument.

Amylifwation/Detection

[0353] Each master mix is designed to amplify and detect 2 or 3 IDHl amino acid mutations (codon with mutant nucleotide underlined). Oligonucleotide 1 master mix amplifies and detects R132C (TGT) and R132H (CAT). Oligonucleotide 2 master mix amplifies and detects R132G (GGT), R132S (AGT), and R132L (CTT). Refer to Table 38. In addition, both master mixes are designed to amplify and detect a region of the IDHl gene outside of codon 132, which serves as an endogenous internal control (IC). Table 38. IDH Mutation Detected by Each Master Mix

[0354] During the amplification reaction on a real-time PCR instrument, the target DNA is amplified by DNA polymerase in the presence of primers, deoxyribonucleoside triphosphates (dNTPs), and magnesium chloride (MgC12). The DNA polymerase used in the assay is a thermophilic enzyme that has been chemically modified, rendering it inactive (e.g., inactive at ambient temperature).

[0355] During the amplification reaction of the IDHl Assay of this Example, DNA polymerase is first activated at a high temperature. During each subsequent round of thermal cycling, a high temperature is used to melt double-stranded DNA strands, followed by a low temperature where primers anneal to their respective targets and are extended to generate double- stranded DNA products. Exponential amplification of the products is achieved through repeated cycling between high and low temperatures. Amplification of IDHl mutation and IC targets takes place simultaneously in the same PCR well.

[0356] IDHl products are detected during the annealing/extension step by measuring the real-time fluorescence signals of the IDHl mutation and IC-specific probes, respectively. The IDHl mutation and IC-specific probes are labeled with different fluorophores, allowing their signals to be distinguishable in a single PCR well.

Assay Protocol

[0357] The IDHl Assay protocol includes the following steps:

A. Manual preparation (i.e., DNA extraction) of samples (specimens and controls).

B. PCR assay setup using the sample eluates and an IDHl amplification reagent kit.

C. Amplification/detection on a real-time PCR instrument.

Assay Results

[0358] For each patient sample, 2 PCR reactions are evaluated. The IDHl Assay of this Example is a qualitative assay for which specimen interpretations are reported as“Mutation Detected” or“Not Detected.” For specimens with interpretations of“Mutation Detected”, the identity of the IDHl mutation detected is reported.

Prevention of Nucleic Acid Contamination

[0359] The possibility of nucleic acid contamination is minimized because:

• IDHl Assay of this Example performs amplification and fluorescence detection in a sealed 96- well optical reaction plate.

• Detection is carried out automatically without the need to open the 96-well optical reaction plate.

• Aerosol barrier pipette tips are used for all pipetting. The pipette tips are discarded after use.

• Separate dedicated areas are used to perform IDHl Assay of this Example.

Claims

CLAIMS We claim:

1. A method of treating a patient diagnosed with a cancer characterized by (i) an IDH-1 mutation selected from R132G, R132S, and R132L and (ii) a mutation selected from DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXL1, FLT3, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53, the method comprising orally administering 150 mg of Compound 1 :

Compound 1

twice daily to the patient in need thereof.

2. The method of claim 1, wherein the cancer is not characterized by an IDH2 mutation.

3. The method of claim 1 or 2, wherein the cancer is characterized by a mutation selected from FLT3, NPM1, CEBPA and TP53.

4. The method of any one of claims 1-3, wherein the cancer is characterized by a mutation selected from DNMT3A, TP53, ATM, and NRAS.

5. The method of any one of claims 1-4, wherein the cancer is acute myeloid leukemia or myelodysplastic syndrome.

6. The method of any one of claims 1-4, wherein the cancer is glioma.

7. The method of any one of claims 1-6, wherein Compound 1 is administered to the patient every day for at least 6 months.

8. The method of any one of claims 1-7, wherein Compound 1 is administered as a single agent.

9. The method of any one of claims 1-7, wherein Compound 1 is administered to the patient in combination with azacitidine during one or more 28-day treatment cycles, wherein:

a. the azacitidine is administered to the patient at the dose of 75 mg/m² for 7 days IV/SC per every 28-day cycle; and

b. a total of 150 mg of Compound 1 is administered BID to the patient every day throughout the one or more 28-day treatment cycles.

10. A method of treating a patient diagnosed with a hematological malignancy characterized by (i) at least one IDHl mutation selected from R132C, R132H, R132G, R132S, and R132L and (ii) at least one mutation selected from CEBPA, DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXL1, FLT3, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53, the method comprising orally administering 150 mg of Compound 1 :

as a single agent twice daily to the patient in need thereof.

11. The method of claim 10, wherein the hematological malignancy is with acute myeloid leukemia (AML) or myelodysplastic syndrome (MDS).

12. A method of treating a patient diagnosed with glioma characterized by (i) at least one IDHl mutation selected from R132C, R132H, R132G, R132S, and R132L and (ii) at least one mutation selected from DNMT3 A, TP53, ATM, and NRAS, the method comprising orally administering 150 mg of Compound 1 :

twice daily to the patient in need thereof.

13. The method of claim 12, wherein Compound 1 is administered as a single agent for treatment of glioma.

14. A method of treating a patient diagnosed with a hematological malignancy characterized by (i) at least one IDHl mutation selected from R132C, R132H, R132G, R132S, and R132L and (ii) at least one mutation selected from CEBPA, DNMT3A, NPM1, SRSF2, NRAS, RUNX1, ASXLl, STAG2, TET2, SMC1A, SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, BCOR, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53, the method comprising orally administering 150 mg of Compound 1 :

twice daily in combination with azacitidine to the patient in need thereof.

15. The method of claim 14, wherein the hematological malignancy is characterized by at least one mutation selected from NPM1, SRSF2, RUNX1, ASXL1, STAG2, TET2, SMC1A,

SF3B1, U2AF1, PHF6, JAK2, MPL, NF1, ASXL2, EED, WT1, CBL, CSF3R, ETNK1, PTPN11, ATM and TP53.

16. A method of treating a patient diagnosed with a glioma characterized by an IDH1 mutation and at least one co-mutation selected from DNMT3A, TP53, ATM, and NRAS, the method comprising orally administering 150 mg of olutasidenib twice daily to the patient in need thereof.

17. The method of claim 16, comprising repeatedly administering the olutasidenib to the patient in need thereof on consecutive days throughout a course of treatment of at least 15 consecutive days.

18. The method of claim 17, wherein the course of treatment is one or more consecutive 28- day treatment cycles.

19. The method of claim 18, wherein the course of treatment is at least 6 consecutive treatment cycles.

20. The method of claim 19, wherein the course of treatment is at least 9 consecutive treatment cycles.