Method for treating cancer with oral dosage form of estrogen receptor-alpha inhibitor

Abstract

To provide pharmaceutical compositions comprising an inhibitor of human ERα, and methods of cancer therapy using the ERα inhibitor.SOLUTION: The present invention provides an oral dosage form comprising i) (E)-NN-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide or a pharmaceutically acceptable salt thereof, and ii) at least one pharmaceutically acceptable excipient, wherein the oral dosage form, when administered orally once daily to a human subject, is formulated to achieve a mean Cmx of about 1 ng / mL to about 4 ng / mL as measured for every mg of formula (I) in the dosage form.SELECTED DRAWING: None

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001]

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 852,751, filed May 24, 2019, and U.S. Provisional Patent Application No. 62 / 853,498, filed May 28, 2019, which are incorporated herein by reference in their entireties. BACKGROUND OF THEINVENTION

[0002]

[0002] Alterations in estrogen receptor (hereinafter ER) signaling are known to play an important role in the development of various types of breast cancer. Activation of ER signaling typically depends on the expression of endogenous steroid hormones, such as estradiol, and their ability to penetrate into cells and interact with intracellular ERs. Once activated, ER signaling promotes various cellular processes, such as proliferation, angiogenesis, metabolism, and cell survival (Toss A.et al.2017,Int.J.Mol.Sci.,18(1):85). During tumorigenesis, it is common for mutations to occur in different ERs to develop, where the mutations confer the cancer cells the ability to activate the ER signaling pathway in a ligand-independent manner. A particular ER known to be associated with breast cancer tumorigenesis, ERα, is a hormone-regulated transcription factor present in 50% of all breast cancers (Lumachi F.,Curr.Med Chem.2013;20:596-604).

[0003]

[0003] Expression of estrogen receptor alpha (ERα, ESR1), a hormone-regulated transcription factor, occurs in approximately 70% of breast cancers. Lumachi F., Curr. Med. Chem. 2013;20:596-604. Numerous anti-ERα therapies have been developed. Resistance to ERα antagonists involves several mechanisms and is common in the clinic. One mechanism, ERα mutations, occurs in 20-40% of endocrine therapy-resistant metastases. Li S, Cell Rep. 2013;4:1116-1130; Robinson DR, Nat Genet. 2013;45:1446-1451; Toy W, Nat Genet. 2013;45:1439-1445; Chandarlapaty S, JAMA Oncol. 2016;2:1310-1315; Spoerke JM, Nat Commun. 2016;7:11579. Mutations in ERα result in ligand-independent activation of the ERα pathway.

[0004]

[0004] Endocrine therapy is the forefront of therapeutic methods used to treat or cure breast cancer. Endocrine therapy is a treatment that involves promoting or inhibiting the expression or function of hormones in the body to treat a particular disease. Endocrine therapy targeting ERα has been shown to be ineffective as a long-term treatment method for breast cancer because 20%-40% of all breast cancers that express ERα acquire mutations after long-term endocrine treatment that render the receptor unresponsive to conventional endocrine treatment (Robinson, DR, Nat. Genet. 2013; 45: 1446-1451). Thus, there is a need in the field of chemotherapy to develop novel therapeutic methods that can effectively target the mutant isoforms of ERα that render cancer resistant to endocrine treatment. In the pursuit of developing new therapeutic approaches for endocrine therapy-resistant breast cancer, a novel class of ERα inhibitors, termed Selective Estrogen Receptor Covalent Antagonists (hereinafter SERCA), was discovered. SERCA inactivates ER signaling by targeting cysteine ​​residues on the ER that are not present in other nuclear hormone receptors (Puyang, X., Cancer Discov. 2018, 8(9):1176-1193). One SERCA that emerged during this period is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, which has the following formula I: [ka] As shown by:

[0005]

[0005] The free base form of (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide may be referred to as H3B-6545. H3B-6545 is a covalent small molecule inhibitor that inactivates both wild-type ERα (ERα-WT) and mutant ERα (ERα-mut) without degrading the receptor. H3B-6545 can interact with the Cys530 residue of ERα, causing the receptor to adopt a unique conformation that inhibits the receptor's ability to promote ligand-independent ERα signaling transduction (Puyang, X., Cancer Discov. 2018, 8(9):1176-1193). H3B-6545 has been shown to exert potent chemotherapeutic properties in various breast cancer cell lines and patient-derived xenograft models in nude mice (Smith, PG, et al., Cancer Res. 2017).

[0006]

[0006] Although H3B-6545 has been shown to be effective in in vitro and in vivo models, it remains to be determined how H3B-6545 should be administered to human breast cancer patients in need of treatment. Thus, there is a need to provide a formulation and administration regimen of the inhibitor that allows H3B-6545 to be administered more effectively and safely to human subjects in need thereof. Characterization of the pharmacokinetic (hereafter PK) outcomes and subsequent PK profile will allow for its general use in human breast cancer treatment methods. Brief Overview

[0007]

[0007] Embodiments may provide, for example, an oral dosage form comprising a compound according to formula I, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient, wherein said formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, said oral dosage form having a mean C of about 1 ng / mL to about 4 ng / mL, as measured for each mg of formula I in said dosage form, when administered once daily to a human subject. max In some embodiments, the average C max is about 2 ng / mL to about 4 ng / mL for each mg of Formula I in the dosage form. max is about 3 ng / mL to about 4 ng / mL for each mg of Formula I in the dosage form. max ranges from 80%-125% of 3 ng / mL to 80%-125% of 3.5 ng / mL for each mg of Formula I in said dosage form.

[0008] In a further embodiment, the dosage form has a mean C max The average of t maxIn a further embodiment, the dosage form is formulated to achieve said mean C in about 3 hours to about 7 hours. max The average of t max In a further embodiment, the dosage form is formulated to achieve said mean C in about 3.5 hours to about 4.5 hours. max The average of t max In a further embodiment, the dosage form is formulated to achieve said mean C in about 5.5 hours to about 6.5 hours. max The average of t max It is formulated to achieve this.

[0009] In some embodiments of the above, the dosage form comprises from about 100 mg to about 600 mg total equivalents of Formula I. In some embodiments, the dosage form comprises about 450 mg total equivalents of Formula I.

[0010] A further embodiment provides an oral dosage form comprising a compound according to formula I, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient, wherein said formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, said oral dosage form having a mean AUC of about 16 h*ng / mL to about 44 h*ng / mL for each mg of formula I in said dosage form, when orally administered once daily to a human subject. 0-24 In some embodiments, the mean AUC 0-24 is about 27 h*ng / mL to about 44 h*ng / mL for each mg of Formula I in the dosage form. 0-24 ranges from 80%-125% of 30h*ng / mL to 80%-125% of 44h*ng / mL for each mg of Formula I in the dosage form. In some embodiments, the dosage form contains a total equivalent of about 100 mg to about 600 mg of Formula I. In some embodiments, the dosage form contains a total equivalent of about 450 mg of Formula I.

[0011] A further embodiment provides an oral dosage form comprising a compound of formula I or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient, wherein the compound of formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, and wherein the oral dosage form, when orally administered once daily to a human subject, has an average t 1 / 2 In a further embodiment, the average t 1 / 2 is about 8 hours to about 12 hours. In a further embodiment, the average t 1 / 2 is about 9 hours to about 11 hours. In some embodiments, the dosage form comprises about 100 mg to about 600 mg total equivalent of Formula I. In some embodiments, the dosage form comprises about 450 mg total equivalent of Formula I.

[0012] A further embodiment provides an oral dosage form comprising a compound of formula I or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient, wherein the compound of formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, wherein the oral dosage form, when orally administered once daily to a human subject, has a mean AUC of about 21 h*ng / mL to about 67 h*ng / mL for each mg of formula I in the dosage form. 0-inf In some embodiments, the compound is formulated to achieve a mean AUC 0-inf is about 29 h*ng / mL to about 67 h*ng / mL for each mg of Formula I in the dosage form. 0-infranges from 80%-125% of 36h*ng / mL to 80%-125% of 57h*ng / mL for each mg of Formula I in the dosage form. In some embodiments, the dosage form contains a total equivalent of about 100 mg to about 600 mg of Formula I. In further embodiments, the dosage form contains a total equivalent of about 450 mg of Formula I.

[0013]

[0013] Still further embodiments may include the dosage forms described herein having means for achieving the pharmacokinetic values ​​described herein.

[0014] A further embodiment provides an oral dosage form comprising a compound of formula I or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient, wherein said compound of formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, said oral dosage form, when orally administered once daily to a human subject, has a mean AUC of about 16 h*ng / mL to about 41 h*ng / mL for each mg of formula I in said dosage form. 0-t In a further embodiment, the mean AUC 0-t ranges from 80%-125% of 27h*ng / mL to 80%-125% of 36h*ng / mL for each mg of Formula I in the dosage form. In a further embodiment, the dosage form contains a total equivalent of about 100 mg to about 600 mg of Formula I. In a further embodiment, the dosage form contains a total equivalent of about 450 mg of Formula I.

[0015] In further embodiments, the previously reported dosage form may be a capsule comprising an internal phase comprising Formula I or a pharma- ceutically acceptable salt, lactose monohydrate, low-substituted hydroxypropylcellulose, microcrystalline cellulose, hydroxypropylcellulose, colloidal anhydrous silica, and magnesium stearate, and an external phase comprising magnesium stearate. In some embodiments, the capsule is a hypromellose capsule. In some embodiments, the capsule comprises the mono-HCl salt form of Formula I.

[0016] In some previously reported embodiments, the oral dosage form is a tablet comprising Formula I or a pharma- ceutically acceptable salt, an inner phase comprising lactose monohydrate, low-substituted hydroxypropylcellulose, hypromellose, colloidal silicon dioxide, and purified water, an outer phase comprising microcrystalline cellulose and magnesium stearate, and a film coating comprising hypromellose, talc, titanium dioxide, propylene glycol, iron oxide, and purified water. In further embodiments, the tablet comprises the mono-HCl salt form of Formula I.

[0017] A further embodiment provides a method of treating cancer in a human subject comprising administering to the subject an oral dosage form comprising a therapeutically effective amount of a compound of formula I, or a pharma- ceutical acceptable salt thereof, and at least one pharma- ceutical acceptable excipient, wherein said compound of formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, said therapeutically effective amount being a single daily dose in the range of about 100 mg to 600 mg, and said oral dosage form having a mean C of about 1 ng / mL to about 4 ng / mL in blood plasma of said subject for each mg of formula I in said dosage form. max In some embodiments, the average C max is about 2 ng / mL to about 4 ng / mL for each mg of Formula I in the dosage form.max is about 3 ng / mL to about 4 ng / mL for each mg of Formula I in the dosage form. max is in the range of 80%-125% of 3 ng / mL to 80%-125% of 3.5 ng / mL for each mg of Formula I in the dosage form. In some embodiments, the dosage form has a C max The average of t max In some embodiments, the dosage form has the average C of formula I of about 3 hours to about 7 hours. max The average of t max In some embodiments, the dosage form has the average C of formula I of about 3.5 hours to about 4.5 hours. max The average of t max In some embodiments, the dosage form has the average C of formula I of about 5.5 hours to about 6.5 hours. max The average of t max has.

[0018] A further embodiment may provide a method of treating cancer in a human subject comprising administering to the subject an oral dosage form comprising a therapeutically effective amount of a compound of formula I, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient, wherein the compound of formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, and wherein the oral dosage form has a mean AUC 0-24 In a further embodiment, the mean AUC 0-24 is about 27 h*ng / mL to about 44 h*ng / mL for each mg of Formula I in the dosage form. 0-24 ranges from 80%-125% of 30 h*ng / mL to 80%-125% of 44 h*ng / mL for each mg of Formula I in said dosage form.

[0019] A further embodiment provides a method of treating cancer in a human subject comprising administering to the subject an oral dosage form comprising a therapeutically effective amount of a compound of formula I or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient, wherein the compound of formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, and the oral dosage form has a mean t 1 / 2 In some embodiments, the average t 1 / 2 In some embodiments, the average t 1 / 2 is about 9 to 11 hours.

[0020] A further embodiment provides a method of treating cancer in a human subject comprising administering to the human subject an oral dosage form comprising a therapeutically effective amount of a compound of formula I or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient, wherein the compound of formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, and wherein the oral dosage form has a mean AUC 0-inf In some embodiments, the mean AUC 0-inf is about 29 h*ng / mL to about 67 h*ng / mL for each mg of Formula I in the dosage form. 0-inf ranges from 80%-125% of 36 h*ng / mL to 80%-125% of 57 h*ng / mL for each mg of Formula I in said dosage form.

[0021] A further embodiment provides a method of treating cancer in a human subject comprising administering to the subject an oral dosage form comprising a therapeutically effective amount of a compound of formula I or a pharma- ceutical acceptable salt thereof and at least one pharma- ceutical acceptable excipient, wherein the compound of formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, and wherein the oral dosage form has a mean AUC 0-t In a further embodiment, the mean AUC 0-t ranges from 80%-125% of 27 h*ng / mL to 80%-125% of 36 h*ng / mL for each mg of Formula I in said dosage form.

[0022] In some embodiments reported herein, the dosage form contains from about 100 mg to about 600 mg total equivalents of Formula I. In other embodiments, the dosage form contains from about 200 mg to about 600 mg total equivalents of Formula I. In other embodiments, the dosage form contains from about 300 mg to about 600 mg total equivalents of Formula I. In other embodiments, the dosage form contains from about 450 mg total equivalents of Formula I.

[0023] In some embodiments reported herein, the oral dosage form is a capsule comprising a compound of formula I or a pharma- ceutically acceptable salt, an internal phase comprising lactose monohydrate, low-substituted hydroxypropylcellulose, microcrystalline cellulose, hydroxypropylcellulose, colloidal anhydrous silica, and magnesium stearate, and an external phase comprising magnesium stearate. In some embodiments, the capsule is a hypromellose capsule. In some embodiments, the capsule comprises the mono-HCl salt form of formula I.

[0024] In some embodiments reported herein, the oral dosage form is a tablet comprising a compound of formula I or a pharma- ceutically acceptable salt, an inner phase comprising lactose monohydrate, low-substituted hydroxypropylcellulose, hypromellose, colloidal silicon dioxide, and purified water, an outer phase comprising microcrystalline cellulose and magnesium stearate, and a film coating comprising hypromellose, talc, titanium dioxide, propylene glycol, iron oxide, and purified water. In some embodiments, the tablet comprises the mono-HCl salt form of formula I.

[0025]

[0025] In some embodiments reported herein, the cancer is breast cancer. In some embodiments, the breast cancer is an ERα-positive breast cancer. In some embodiments, the breast cancer expresses wild-type ERα. In some embodiments, the breast cancer expresses mutant ERα.

[0026] In some embodiments reported herein, the oral dosage form is administered once daily. In some embodiments, the oral dosage form is administered to a human in a fasting state. In some embodiments, the oral dosage form is administered to a human in a fed state.

[0027]

[0027] An embodiment is the compound H3B-6545:

[0028]

[0028] [ka] (H3B-6545)

[0029] or a pharmaceutically acceptable salt thereof, comprising the steps of: determining whether one or more decreases in mutant allele frequency are observed in the patient's blood; selecting the patient if said decreases are observed; and administering to the selected patient a therapeutically effective amount of H3B-6545 or a pharmaceutically acceptable salt thereof. Mutant allele frequency is the relative frequency of a mutant allele (variant of a gene) at a particular locus in a population, expressed as a fraction or percentage ratio.

[0030]

[0030] H3B-6545 is reported in US Pat. No. 9,796,683, which is incorporated herein by reference.

[0031]

[0031] In some embodiments reported herein, the pharma ceutically acceptable salt of H3B-6545 is the HCl salt of H3B-6545.

[0032]

[0032] In some embodiments, a decrease in mutant allele frequency is observed in one or more alleles selected from the group consisting of PIK3CA, ESR1, TSC1, TP53, FGFR1, CCND1, ARID1A, POLE, FGF19, MET, NOTCH3, FGF3, AKT1, KRAS, MYC, ERBB2, ERBB3, FGFR3, PMS2, PTEN, RB1, BRAF, MDM2, ATR, ATRX, BRCA2, SETD2, ATM, FANCA, JAK2, NF1, SLX4, and SMAD4.

[0033] In a further embodiment, a decrease in mutant allele frequency is observed in AKT1. In a further embodiment, the AKT1 mutation position that results in the observed decrease in mutant allele frequency is E17. In a further embodiment, the AKT1 mutation that results in the observed decrease in mutant allele frequency is E17K.

[0034]

[0034] In a further embodiment, the reduction in mutant allele frequency is observed in ESR1. In yet a further embodiment, the ESR1 mutation position that results in the observed reduction in mutant allele frequency is selected from one or more members of the group consisting of E380, D538, L536, S463, and Y537. In a further embodiment, the ESR1 mutation that results in the observed reduction in mutant allele frequency is selected from one or more members of the group consisting of E380Q, D538G, L536H, L536P, L536R, S463P, Y537C, Y537N, and Y537S. In a further embodiment, the ESR1 mutation that results in the observed reduction in mutant allele frequency is S463P. In a further embodiment, the ESR1 mutation that results in the observed reduction in mutant allele frequency is Y537N.

[0035] In a further embodiment, the reduction in mutant allele frequency is observed in PIK3CA. In a further embodiment, the PIK3CA mutation position that results in the observed reduction in mutant allele frequency is selected from one or more members of the group consisting of E542, E545, and H1047. In a further embodiment, the PIK3CA mutation that results in the observed reduction in mutant allele frequency is selected from one or more members of the group consisting of E542K, E545K, H1047L, and H1047R.

[0036] A further embodiment is the compound H3B-6545

[0037]

[0037] [ka] (H3B-6545)

[0038]

[0038] A method for stopping treatment with compound H3B-6545 or a pharma- ceutically acceptable salt thereof in a patient receiving treatment with compound H3B-6545 or a pharma- ceutically acceptable salt thereof may be provided, the method comprising determining whether one or more increases in mutant allele frequency are observed in the patient's blood, and if said increases are observed, stopping treatment of the patient.

[0039]

[0039] In some embodiments, an increase in mutant allele frequency is observed in one or more alleles selected from PIK3CA, ESR1, TSC1, TP53, FGFR1, CCND1, ARID1A, POLE, FGF19, MET, NOTCH3, FGF3, AKT1, KRAS, MYC, ERBB2, ERBB3, FGFR3, PMS2, PTEN, RB1, BRAF, MDM2, ATR, ATRX, BRCA2, SETD2, ATM, FANCA, JAK2, NF1, SLX4, and SMAD4.

[0040] In some embodiments, an increase in mutant allele frequency is observed in ESR1. In still further embodiments, the ESR1 mutation position that results in the observed increase in mutant allele frequency is selected from one or more members selected from the group consisting of E380, D538, L536, S463, and Y537. In further embodiments, the ESR1 mutation that results in the observed increase in mutant allele frequency is selected from at least one member selected from the group consisting of E380Q, D538G, L536H, L536P, L536R, S463P, Y537C, Y537N, and Y537S.

[0041]

[0041] In a further embodiment, an increase in mutant allele frequency is observed in PIK3CA. In yet a further embodiment, the PIK3CA mutation position resulting in the observed increase in mutant allele frequency is selected from the group consisting of E542, E545, and H1047. In an even further embodiment, the PIK3CA mutation resulting in the observed increase in mutant allele frequency is selected from the group consisting of E542K, E545K, H1047L, and H1047R. In a further embodiment, the PIK3CA mutation is E545K.

[0042] In a further embodiment, the increased mutant allele frequency is observed in AKT1. In a further embodiment, the AKT1 mutation position that results in the observed increased mutant allele frequency is E17. In a further embodiment, the AKT1 mutation that results in the observed increased mutant allele frequency is E17K.

[0043] In still further embodiments, the identifying step comprises determining whether the patient is a patient receiving the compound H3B-6545: [ka] (H3B-6545)

[0044]

[0044] or a pharma- ceutically acceptable salt thereof.

[0045]

[0045] In a further embodiment, the patient has breast cancer. In yet a further embodiment, the breast cancer is estrogen receptor alpha (ERα) positive breast cancer. In yet an even further embodiment, the ERα is mutated.

[0046] In yet a further embodiment, an elevated mutant allele frequency is observed in circulating tumor DNA. In a further embodiment, a decreased mutant allele frequency is observed in circulating tumor DNA.

[0047]

[0047] The documents reported in this specification are incorporated by reference. In case these documents conflict with this document, this document shall take precedence. [Brief description of the drawings]

[0048] [Figure 1] Figure 1 shows the mean plasma concentration profile over time for H3B-6545 (Cycle 1, Day 1) in Example 3. Preliminary PK analysis shows that H3B-6545 exhibits a roughly dose-proportional increase in plasma exposure from 100 mg to 600 mg. tmax and half-life appeared to be dose and time independent. [Diagram 2] Figure 1 shows tumor response and duration of H3B-6545 treatment as reported in Example 3. Partial response, stable response, progressive disease, and non-evaluable are indicated for each tumor assessment. Arrows represent ongoing treatment at the time of data cut. [Diagram 3] 4 shows the percentage change in the sum of the diameters of the target lesions reported in Example 3. [Figure 4] FIG. 1 shows a schematic diagram of liquid biopsy and tissue biopsy sample sets. [Diagram 5] 5A-5E show baseline genomics profiles of patients in Example 7. BEAMing assay: Fig. 5A. Baseline ESR1 and PIK3CA mutation status in patients, Fig. 5B and Fig. 5C. Clonality of ESR1 and PIK3CA mutations, Fig. 5D and Fig. 5E. Amino acid distribution of PIK3CA and ESR1 mutations. [Figure 5F] 1 shows the oncoprint of mutations found in baseline tissue biopsies as determined by the Oncomine comprehensive panel. [Figure 6A-6B] Figure 6 shows the concordance of mutations found in tissue and liquid biopsies in Example 7. Figure 6A shows the allele frequency distribution and concordance of tissue and liquid biopsy mutations. Figure 6B is a summary table of the concordance of tissue and liquid biopsy mutations. [Figure 7A-7C]Figure 7 shows ctDNA kinetics correlated with overall best response to H3B-6545. AF=allele frequency, C=cycle, D=day, PD=progressive disease, PR=partial response, SD=stable disease. Dotted lines represent ddPCR assay LOD. Figure 7A shows the ratio of AF baseline / C2D1 for each patient colored individually. A patient may have multiple mutations. Figures 7B and 7C show exemplary ctDNA kinetics in patients with progressive disease (B) vs partial response (C). Detailed Description

[0049]

[0055] definition

[0050]

[0056] The use of the articles "a," "an," and "the" herein is intended to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. For example, the term "a disintegrant" refers to one or more disintegrants contained in or suitable for use in the formulations described herein. Similarly, the term "a therapeutic amount" refers to one or more therapeutic amounts contained in or suitable for use in the dosage form.

[0051]

[0057] The terms "comprising," "having," "including," and "containing" are open-ended (i.e., meaning "including, but not limited to"), unless otherwise stated. Furthermore, whenever "comprising" or other open-ended terminology is used in an embodiment, it is to be understood that the same embodiment may also be claimed more narrowly using the intermediate term "consisting essentially of" or the closed term "consisting of."

[0052]

[0058] The expressions "bioequivalent" or "bioequivalence" are technical terms and are intended to be defined according to Approved Drug Products with Therapeutic Equivalence Evaluations, 34th edition, published by the United States Department of Health and Human Services, commonly known as the "Orange Book". Bioequivalence of different formulations of the same drug substance involves equivalence with respect to the rate and extent of drug absorption. The extent and rate of absorption of the test formulation is compared to the reference formulation to determine whether the two formulations are bioequivalent. Standard bioequivalence studies are performed in a crossover fashion with extensive testing involving administering single doses of the test and reference drugs to a number of volunteers, usually 12-24 healthy normal adults, and then measuring blood or plasma levels of the drug over time. Detailed guidelines for establishing the bioequivalence of a drug formulation with a reference formulation are published by the FDA Office of Generic Drugs, Division of Bioequivalence.

[0053]

[0059] As used herein, the term "mean" refers to the geometric mean determined from a set of independent measurements. For example, the independent measurements may be collected from a statistically significant population. As a further example, when used to describe a pharmacokinetic parameter (e.g., "mean C max ”, “Average AUC 0-x ”, “Average AUC 0-t ”, “Average AUC 0-inf ”, “Average t max " or "Average t 1 / 2 (or "mean half-life")), "mean" refers to the geometric mean pharmacokinetic value obtained from a population from which each individual measurement was collected. Thus, as used herein, a dosage form may be administered to a human subject, wherein the dosage form has an average pharmacokinetic value obtained from a collection of independently measured values.

[0054]

[0060] The list of abbreviations and definitions of terms used in this application is as follows: AUC: Area under the plasma concentration-time curve; AUC 0-x : Area under the plasma concentration-time curve from zero to x hours after administration (e.g., x can represent 12 hours or 24 hours), AUC 0-t : Area under the plasma concentration-time curve from time zero to the last quantifiable concentration, AUC 0-inf : Area under the plasma concentration-time curve from time zero to infinity, ANCOVA: Analysis of variance, CI: Confidence interval, C max : Maximum drug concentration, C x : Plasma concentration at x hours after administration, CV: Coefficient of variance, LC-MS / MS: Liquid chromatography mass spectrometry / mass spectrometry, MAD: Multiple ascending dose study, MTD: Maximum tolerated dose, PD: Pharmacodynamics, PK: Pharmacokinetics, RT: Response time, SAD: Single ascending dose study, SD: Standard deviation, t 1 / 2 : Terminal elimination half-life, t max t: Time to maximum (peak) concentration after drug administration. As used herein, t 1 / 2 contains the terminal elimination half-life of the drug concentration, which is C max As used herein, C may be a terminal elimination half-life of max contains the maximum drug concentration of the substance measured in human plasma.

[0055]

[0061] Two dosage forms whose rate and extent of absorption differ by less than -20% / +25% are generally considered to be "bioequivalent". Another approach for mean bioequivalence involves the calculation of a 90% confidence interval for the ratio of the means (population geometric means) of measurements for the test product and the reference product. To establish BE, the calculated confidence interval should usually be within 80-125% for the ratio of the product means. In addition to this general approach, other approaches, such as (1) logarithmic transformation of pharmacokinetic data, (2) methods for assessing sequence effects, and (3) methods for assessing outliers, may be useful for establishing bioequivalence. For example, in (1) above, the confidence interval should usually be within 80-125% for the difference in the mean values ​​of the logarithmically transformed PK parameters.

[0056]

[0062] As used herein, the terms "about," "approximately," or "approximate," when referring to measurable values, such as amounts, time periods, and the like, are meant to include a ±20% or 10% variation from the particular value, more preferably a 5%, even more preferably a 1%, and even more preferably a 0.1% variation, as such variation is appropriate in a given context.

[0057]

[0063] When a dose or dose range is referred to by mass or weight in the format of "compound of formula I" or "H3B-6545" or "free base of H3B-6545", one skilled in the art will understand that providing an equivalent molar amount of an active compound as a pharma- ceutically acceptable salt typically requires administration of a greater mass of salt than administration of the compound alone (i.e., the amount of free base and the amount of salt have a 1:1 molar ratio). For example, the phrase "25 mg to 50 mg of a compound of formula I or a pharma- ceutically acceptable salt thereof" contemplates an amount of 25 mg to 50 mg (inclusive of the endpoint) of the free base of the compound of formula I, as well as an amount of 26 mg to 53 mg (inclusive of the endpoint) of the monohydrochloride salt of the compound of formula I. This conversion may be referred to, for example, as a "salt conversion factor", "salt correction factor" or "potency adjustment factor".

[0058]

[0064] The potency adjustment factor conversion is also applicable to crystalline forms of Formula I that exist as hydrates, solvates, or crystalline forms of Formula I with both i) hydrates or solvates and ii) salt counterions. Furthermore, such potency adjustment factor conversion is also applicable whether the co-crystal solvent molecules and / or salt counterions are present in the crystalline form in integral or non-integral stoichiometric ratios. Thus, one skilled in the art will appreciate that different potency adjustments can be made for the monohydrochloride salt of H3B-6545, the hemihydrochloride salt of H3B-6545, or unusual ratios such as 1:1.3, 1:1.25, etc.

[0059]

[0065] Consistent with the above paragraph, as used herein, a "corresponding" amount (e.g., mass, weight, dose, etc.) of Formula I (or H3B-6545, the free base of H3B-6545, or any other synonym thereof as used herein) refers to the amount of any salt and / or hydrate after potency adjustment factors.

[0060]

[0066] "H3B-6545 drug substance" refers to the monohydrochloride salt of H3B-6545, as reported in US Pat. No. 10,640,483, which is incorporated herein by reference.

[0061]

[0067] As used herein, "human subject" is interchangeable with "human subject in need of treatment" or "human subject in need thereof," all of which refer to a human subject having breast cancer or at high risk of developing breast cancer relative to the general population. A human subject in need thereof may be a human subject that has previously been diagnosed or identified as having breast cancer or a precancerous condition. Alternatively, a human subject in need thereof may be a human subject at high risk of developing such a disorder relative to the general population (i.e., a subject susceptible to developing such a disorder relative to the general population). A human subject in need thereof may have a precancerous condition.

[0062]

[0068] The human subject in need thereof may have a refractory or resistant cancer (i.e., a cancer that does not respond or has never responded to treatment). The subject may be resistant at the start of treatment or may become resistant during treatment. In some embodiments, the subject in need thereof has a recurrence of cancer after remission during the most recent treatment. In some embodiments, the subject in need thereof has undergone and failed all known effective therapies for the treatment of cancer. In some embodiments, the subject in need thereof has undergone at least one prior treatment. In a preferred embodiment, the subject has cancer or a cancerous condition.

[0063]

[0069] As used herein, "fasted state" refers to a human subject in need of fasting, who has undergone an overnight fast of at least 10 hours prior to administration. No food should be allowed for at least 4 hours after administration. Water may be allowed except for 1 hour before and after drug administration.

[0064]

[0070] As used herein, "fed state" refers to human subjects who received a single dose of H3B-6545 within 30 minutes of the start of a meal after an overnight fast of at least 10 hours and after consuming a high-fat breakfast within 30 minutes. Water was allowed as desired, except for 1 hour before and after drug administration.

[0065]

[0071] As used herein, "treating" or "treat" refers to the management and care of a human subject for the purpose of combating a disease, condition or disorder, and includes the administration of a dosage form of H3B-6545 or a pharma- ceutically acceptable salt, polymorph, hydrate or solvate thereof to alleviate the symptoms or complications of a disease, condition or disorder, or to eliminate such disease, condition or disorder.

[0066]

[0072] H3B-6545 or a pharma- ceutically acceptable salt and / or solvate thereof can or may be used to prevent the associated disease, condition or disorder, or to identify suitable candidates for such purposes. As used herein, "preventing," "prevent," or "protecting against" refers to reducing or eliminating the onset of symptoms or complications of such disease, condition, or disorder.

[0067]

[0073] As used herein, "sample" refers to any biological sample from a human subject, including, but not limited to, cells, tissue samples, bodily fluids (including, but not limited to, mucus, blood, plasma, serum, urine, saliva, and semen), tumor cells, and tumor tissue. Preferably, the sample is selected from bone marrow, peripheral blood cells, blood, plasma, and serum. The sample may be provided by the subject under treatment or examination. Alternatively, the sample may be obtained by a physician, according to routine practice in the art.

[0068]

[0074] As used herein, the term "dosage form" refers to a physically discrete unit suitable as a unitary dose for a human subject to be treated, each unit containing a predetermined amount of active compound calculated to produce a desired therapeutic effect in association with the necessary pharmaceutical carrier. Dosage forms are classified in terms of the route of administration and the site of application, including, for example, oral, topical, rectal, vaginal, intravenous, subcutaneous, intramuscular, ocular, nasal, ocular, and inhalation administration. Alternatively, dosage forms are classified in terms of the physical form, such as, for example, solid, semi-solid, or liquid. Dosage forms may be any of a variety of forms, including, for example, an IV bag, a tablet, a single pump on an aerosol inhaler, or a vial. "Oral dosage form" refers to a dosage form that is easily administered to a human subject through the mouth. Non-limiting examples of oral dosage forms include capsules and tablets. The amount of active ingredient in a unit dose (e.g., a formulation of the disclosed compound or its salt, hydrate, or solvate) is an effective amount and varies according to the particular treatment involved.

[0069]

[0075] As used herein, the phrase "pharmacologically acceptable" refers to compounds, materials, compositions, carriers and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0070]

[0076] As used herein, "a pharma- ceutically acceptable excipient" means an excipient useful for the preparation of a pharmaceutical composition that is generally safe, non-toxic, and not biologically or otherwise undesirable, and includes excipients acceptable for veterinary use as well as for human pharmaceutical use. As used in the specification and claims, "a pharma- ceutically acceptable excipient" includes both one and more than one such excipient. For example, a pharma- ceutically acceptable excipient used in the formulation of the present invention may be a diluent or inert carrier, a lubricant, a binder, or a combination thereof. A pharma- ceutically acceptable excipient used in the formulation of the present invention may further include a filler, an antimicrobial agent, an antioxidant, an anti-caking agent, a coating agent, or a mixture thereof.

[0071]

[0077] The term "composition" as used herein includes a product that includes a specific amount of a specific component and any product that results directly or indirectly from the combination of the specific amount of the specific component. Such a term with respect to pharmaceutical composition is intended to include a product that includes an active ingredient (herein, any of formula I or its pharma-ceutically acceptable salts, hydrates, and / or solvates) and an inactive ingredient that constitutes a carrier, and to include any product that results directly or indirectly from the combination, complexation or aggregation of any two or more components, or the dissociation, other types of reaction or interaction of one or more components. Thus, the pharmaceutical composition of the present invention includes any composition prepared by mixing a compound represented by formula I (or a pharma-ceutically acceptable salt, hydrate, and / or solvate) with a pharma-ceutically acceptable excipient.

[0072]

[0078] As used herein, the term "therapeutically effective amount" refers to an amount of H3B-6545 that can produce a therapeutic effect in a human subject. A therapeutically effective amount is an amount that can treat, ameliorate, or prevent a specified disease or condition, or that exhibits a detectable therapeutic or inhibitory effect. The effect can be detected by any assay method known in the art. The exact effective amount for a subject will depend on the subject's weight, size, and health, the nature and extent of the condition, and the treatment selected for administration. The therapeutically effective amount for a given situation can be determined by routine experimental methods that are within the skill and judgment of the clinician.

[0073]

[0079] In a preferred embodiment, the disease or condition to be treated is cancer. In another embodiment, the disease or condition to be treated is a cell proliferation disorder. The therapeutically effective amount of H3B-6545 can be administered in a dosage form. The therapeutically effective amount of H3B-6545 can be in the form of a pharmaceutically acceptable salt, solvate, and / or hydrate.

[0074]

[0080] As used herein, a "therapeutic effect" is the result of any type of medical treatment, the result of which is deemed desirable and beneficial. This is true whether the result is an expected, unexpected, or even unintended outcome of the treatment. The desirable or beneficial result may be inhibition of alterations in cell signaling pathways, inhibition of cell proliferation, preferably cancer cell proliferation, promotion of cell death, preferably cancer cell death, or shrinkage of the tumor, all of which are observed without severe adverse effects. Slowing tumor growth, and preferably causing regression, and also preferably complete regression of the cancer, are other examples of therapeutic effects. Therapeutic effects can also be an objectively identifiable improvement noted by a clinician or other qualified observer. For example, tumor regression in a patient may be measured with reference to the diameter of the tumor. A decrease in the diameter of the tumor indicates regression. Regression is also indicated by the inability of the tumor to recur after treatment has stopped.

[0075]

[0081] Severe adverse effects include those that are life-threatening (e.g., liver failure, abnormal heart rhythms, and certain types of allergic reactions), result in persistent or significant disability or hospitalization, or cause birth defects.

[0076]

[0082] As used herein, "pharmaceutically acceptable salts" refers to derivatives of H3B-6545, in which the parent compound is modified by making its acid or base salt. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues, such as amines, alkali or organic salts of acidic residues, such as carboxylic acids, and the like. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts or tertiary ammonium salts of the parent compound, formed from non-toxic inorganic or organic acids.

[0077]

[0083] As used herein, "quantifiable" means capable of being measured, calculated or expressed as a quantity or numerical value. A quantifiable plasma concentration is the concentration of H3B-6545 that can be detected and measured in the plasma of a human subject after administration. A quantifiable AUC bioavailability is the fraction of H3B-6545 that can access the systemic circulation of a human subject, which can be calculated by analyzing the plasma concentration of H3B-6545 in plasma samples taken from a human subject over a set period of time. A quantifiable half-life is the detectable or calculated time during which the plasma concentration of H3B-6545 is 50% of the maximum quantifiable plasma concentration of H3B-6545. The methods and materials required to quantify the above-mentioned PK parameters are generally known to those skilled in the art. Specific quantification methods are presented within this application.

[0078]

[0084] As used herein, the term "solvate" refers to a solvent addition form that contains either stoichiometric or non-stoichiometric amounts of solvent.

[0079]

[0085] As used herein, the term "tautomer" means one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to the other.

[0080]

[0086] As used herein, unless specifically stated otherwise or clear from the context, the term "or" is understood to be inclusive.

[0081]

[0087] One aspect of the present invention provides a dosage form that can be administered to a human subject in need thereof, comprising a therapeutically effective amount of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable carrier or excipient, wherein said therapeutically effective amount achieves a quantifiable plasma concentration after administration. In one embodiment of the present invention, the dosage form is an oral dosage form. In another embodiment of the present invention, the dosage form is a solid dosage form. In another embodiment of the present invention, the dosage form is a solid oral dosage form. In yet another embodiment, the solid oral dosage form can be an immediate release oral solid dosage form. The oral solid dosage form can be in the form of a tablet or capsule. These forms can have multiple phases, including, for example, an internal phase and an external phase.

[0082]

[0088] In one embodiment, the dosage form is substantially free of water. In this context, "substantially" free of water means that the water content of the formulation as packaged is less than 7%, less than 5%, less than 1%, or less than 0.5% of the total weight of the formulation. In one embodiment, the amount of water is 0.1-5% (e.g., 0.1-1%, or 0.1-0.5%) of the total weight of the formulation. In one embodiment, the amount of water in the formulation of the present invention produced via a spray coating process is less than 0.5%.

[0083]

[0089] The at least one pharma- ceutically acceptable excipient may be a diluent or inert carrier, a disintegrant, a lubricant, a binder, or a combination thereof. Pharmaceutically acceptable excipients may also include fillers, antimicrobial agents, antioxidants, anti-caking agents, coating agents, or mixtures thereof.

[0084]

[0090] Exemplary binders include corn starch, potato starch, other starches, gelatin, natural and synthetic gums such as acacia, xanthan, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum, cellulose and its derivatives (e.g., ethyl cellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone (e.g., povidone, crospovidone, copovidone, etc.), methylcellulose, methocel, pregelatinized starch (e.g., STARCH 1500® and STARCH 1500 LM® sold by Colorcon, Ltd.), hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose (FMC Corporation, Marcus Hook, PA, USA), Emdex, Plasdone or mixtures thereof; fillers such as talc, calcium carbonate (e.g., granules or powder), dibasic calcium phosphate, tribasic calcium phosphate, calcium sulfate (e.g., granules or powder), microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, dextrose, fructose, honey, lactose anhydrous, lactose monohydrate, lactose and aspartame, lactose and cellulose, lactose and microcrystalline cellulose, maltodextrin, maltose, mannitol, microcrystalline cellulose; and guar gum, molasses, sucrose, or mixtures thereof.

[0085]

[0091] Exemplary disintegrants include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate (e.g., Explotab), potato or tapioca starch, other starches, pregelatinized starch, clays, other algins, other celluloses, gums (e.g., gellan gum), low-substituted hydroxypropylcellulose, ployplasdone, or mixtures thereof.

[0086]

[0092] Exemplary lubricants include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, compritol, stearic acid, sodium lauryl sulfate, sodium stearyl fumarate (e.g., Pruv), vegetable-based fatty acid lubricants, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, com oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, syloid silica gel (AEROSIL 200, W. R. Grace Co., Baltimore, MD USA), coagulated aerosol of synthetic silica (Deaussa Co., Piano, TX USA), pyrogenic silicon dioxide (CAB-O-SIL, Cabot Co., Boston, MA USA), or mixtures thereof.

[0087]

[0093] Exemplary coating agents may include, but are not limited to, sodium carboxymethylcellulose, cellulose acetate phthalate, ethylcellulose, gelatin, pharmaceutical glazes, hydroxypropylcellulose, hydroxypropylmethylcellulose (hypromellose), hydroxypropylmethylcellulose phthalate, methylcellulose, polyethylene glycol, polyvinyl acetate phthalate, shellac, sucrose, titanium dioxide, camauba wax, microcrystalline wax, gellan gum, maltodextrin, methacrylates, microcrystalline cellulose, and carrageenan, or mixtures thereof.

[0088]

[0094] In one embodiment, the dosage form is a solid oral dosage form that can be optionally treated with a coating system (e.g., Opadry® fx film coating system), for example, coated with Opadry® Blue (OY-LS-20921), Opadry® White (YS-2-7063), Opadry® White (YS-1-7040), and Black Ink (S-1-8106).

[0089]

[0095] In one embodiment, the oral dosage form is configured in a capsule having an internal phase comprising a therapeutically effective amount of H3B-6545 or a pharma- ceutically acceptable salt thereof, lactose monohydrate, low-substituted hydroxypropylcellulose, microcrystalline cellulose, hydroxypropylcellulose, and colloidal anhydrous silica, and the capsule also has an external phase comprising magnesium stearate.

[0090]

[0097] In another embodiment, the capsule is hypromellose and further comprises red iron oxide and titanium dioxide.

[0091]

[0098] In another embodiment, the oral dosage form is a capsule comprising, by total weight of the capsule, 25%-30% therapeutically effective amount of H3B-6545 or a pharma- ceutically acceptable salt, 10%-15% lactose monohydrate, 5%-10% low-substituted hydroxypropylcellulose, 1%-5% microcrystalline cellulose, 0.5%-5% hydroxypropylcellulose, 0.05%-0.5% colloidal anhydrous silica, 0.1%-1% magnesium stearate, 40%-45% hypromellose, 0.5%-2% red iron oxide, and 0.5%-2% titanium dioxide.

[0092]

[0099] Pharmaceutically acceptable salts can include the conventional non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids, or the quaternary ammonium salts, such as 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, bicarbonate, carbonic acid, citric acid, edetic acid, ethanedisulfonic acid, 1,2-ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, glycolyarsanilic acid, hexylresorcinic acid, hydrabamic acid, hydrobromic acid, hydrochloric acid, hydroiodic acid, hydroxymaleic acid, hydroxynaphthoic acid, isethionic acid, lactic acid, Included are, but are not limited to, salts derived from inorganic and organic acids selected from lactobionic acid, laurylsulfonic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, napsylic acid, nitric acid, oxalic acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, polygalacturonic acid, propionic acid, salicylic acid, stearic acid, subacetic acid, succinic acid, sulfamic acid, sulfanilic acid, sulfuric acid, tannic acid, tartaric acid, toluenesulfonic acid, and commonly obtained amino acids, such as glycine, alanine, phenylalanine, arginine, and the like.

[0093]

[0100] Other examples of pharma- ceutically acceptable salts include hexanoic acid, cyclopentanepropionic acid, pyruvic acid, malonic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo-[2.2.2]-oct-2-ene-1-carboxylic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, and muconic acid. The present invention also includes salts formed when acidic protons present in the parent compound are replaced by metal ions, e.g., alkali metal ions, alkaline earth ions, or ammonium ions, or coordinate with organic bases, such as ethanolamine, diethanolamine, triethanolamine, tromethamine, and N-methylglucamine. In the salt form, the ratio of compound to cation or anion of the salt can be 1:1 or any ratio other than 1:1, e.g., 3:1, 2:1, 1:2, or 1:3.

[0094]

[0101] In other embodiments, dosage forms may also include one or more active compounds (eg, H3B-6545 or a salt thereof) in combination with at least one pharma- ceutically acceptable excipient or carrier.

[0095]

[0102] Examples of solvates may include that when the solvent is water, the solvate formed is a hydrate, and when the solvent is alcohol, the solvate formed is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one molecule of a substance in which water maintains its molecular state as HO. Hemihydrates are formed by the combination of one molecule of water with more than one molecule of a substance in which water maintains its molecular state as HO.

[0096]

[0103] An oral dosage form comprising a therapeutically effective amount of H3B-6545, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier or excipient, which may be administered to a human subject in need thereof, may achieve a quantifiable plasma concentration from about 0.5 hours to about 12 hours, from about 0.5 hours to about 5 hours, from about 0.5 hours to about 3 hours, from about 1 hour to about 5 hours, or from about 1 hour to about 3 hours after administration.

[0097]

[0104] A therapeutically effective amount of H3B-6545, or a pharma- ceutically acceptable salt thereof, that may be present in an oral dosage form is in the range of about 50 mg to about 1000 mg, about 100 mg to about 800 mg, about 100 mg to about 600 mg, about 200 mg to about 600 mg, or about 400 mg to about 600 mg.

[0098]

[0105] In a preferred embodiment, the therapeutically effective amount of H3B-6545, or a pharma- ceutically acceptable salt thereof, that may be present in an oral dose is 450 mg.In one embodiment of the present invention, the above-mentioned dose is a daily dose.

[0099]

[0106] An oral dosage form comprising a therapeutically effective amount of H3B-6545, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier or excipient, which may be administered to a human subject in need thereof, may achieve a maximum quantifiable plasma concentration, following administration, of about 170 ng / mL to about 2000 ng / mL, about 200 ng / mL to about 1500 ng / mL, about 500 ng / mL to about 2000 ng / mL, about 500 ng / mL to about 1500 ng / mL, about 1000 ng / mL to about 2000 ng / mL, or about 1000 ng / mL to about 1500 ng / mL.

[0100]

[0107] An oral dosage form comprising a therapeutically effective amount of H3B-6545, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier or excipient, which may be administered to a human subject in need thereof, may achieve a half-life of about 5 hours to about 24 hours, about 8 hours to about 24 hours, about 8 hours to about 15 hours, about 10 hours to about 15 hours, or about 15 hours to about 24 hours after administration.

[0101]

[0108] An oral dosage form comprising a therapeutically effective amount of H3B-6545, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier or excipient, which may be administered to a human subject in need thereof, may achieve a quantifiable AUC bioavailability, following administration, of about 2,000 ng*hr / mL to about 20,000 ng*hr / mL, about 5,000 ng*hr / mL to about 20,000 ng*hr / mL, about 10,000 ng*hr / mL to about 20,000 ng*hr / mL, about 15,000 ng*hr / mL to about 20,000 ng*hr / mL, about 5,000 ng*hr / mL to about 15,000 ng*hr / mL, or about 10,000 ng*hr / mL to about 15,000 ng*hr / mL.

[0102]

[0109] An oral dosage form comprising a therapeutically effective amount of H3B-6545, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier or excipient, which may be administered to a human subject in need thereof, may be orally administered as a single dose once daily for a 20-day cycle, a 21-day cycle, a 22-day cycle, a 23-day cycle, a 24-day cycle, a 25-day cycle, a 26-day cycle, a 27-day cycle, a 28-day cycle, or until a therapeutic effect occurs in the subject in need thereof. The dosage form may also be administered in cycles of less than 20 days.

[0103]

[0110] A. An oral dosage form comprising about 100 mg to about 600 mg of H3B-6545, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient.

[0104]

[0111] In some embodiments, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient. In such embodiments, the dose provides a maximum quantifiable plasma concentration C of about 150 ng / mL to about 2100 ng / mL following administration to a subject in need of treatment. max(ng / mL). In other embodiments, the dose achieves a maximum quantifiable plasma concentration after administration to a subject in need of treatment of about 250 ng / mL to about 1700 ng / mL. In further embodiments, the dose achieves a maximum quantifiable plasma concentration after administration to a subject in need of treatment of about 1200 ng / mL to about 1500 ng / mL. In these embodiments, the C max may be accomplished in a time period of about 1 hour to about 25 hours, about 2 hours to about 12 hours, about 2 hours to about 5 hours, or about 4 hours.

[0105]

[0112] In one embodiment, the oral dosage form comprises about 100-600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient. In such an embodiment, the oral dosage form has a C of about 1.8 ng / mL to about 4.0 ng / mL per mg of H3B-6545. max In a further embodiment, the oral dosage form can achieve a C of about 1.8 ng / mL to about 3.4 ng / mL per mg of H3B-6545. max In yet a further embodiment, the oral dosage form can achieve a C of about 3.0 ng / mL to about 4.0 ng / mL per mg of H3B-6545. max A range can be achieved.

[0106]

[0113] In some embodiments, the oral dosage form comprises about 100 mg to 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient. In some of these embodiments, the dosage form achieves a half-life of about 8.0 hours to about 22.0 hours, or a half-life of about 10.0 hours to about 13.0 hours, after administration to a human subject in need thereof.

[0107]

[0114] In one embodiment, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0-24 hr) of about 1,600 ng*hr / mL to about 23,000 ng*hr / mL, about 2,500 ng*hr / mL to about 18,500 ng*hr / mL, about 4,000 ng*hr / mL to about 20,000 ng*hr / mL, or about 9,000 ng*hr / mL to about 17,000 ng*hr / mL after administration to a human subject in need thereof.

[0108]

[0115] In one embodiment, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0-24 hr) of about 17 ng*hr / mL / mg to about 40 ng*hr / mL / mg, about 17 ng*hr / mL / mg to about 37 ng*hr / mL / mg, or about 26 ng*hr / mL / mg to about 40 ng*hr / mL / mg, measured on a "per mg of H3B-6545" basis, following administration to a human subject in need thereof.

[0109]

[0116] In one embodiment, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0 to infinity time) of about 2,000 ng*hr / mL to about 40,000 ng*hr / mL, about 5,000 ng*hr / mL to about 25,000 ng*hr / mL, or about 20,000 ng*hr / mL to about 23,000 ng*hr / mL after administration to a human subject in need thereof.

[0110]

[0117] In one embodiment, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0 to infinity) of about 21 ng*hr / mL / mg to about 66 ng*hr / mL / mg, about 21 ng*hr / mL / mg to about 55 ng*hr / mL / mg, or about 30 ng*hr / mL / mg to about 66 ng*hr / mL / mg, measured on a "per mg of H3B-6545" basis, after administration to a human subject in need thereof.

[0111]

[0118] B. An oral dosage form comprising about 300 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient.

[0112]

[0119] In some embodiments, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient. In such embodiments, the dose provides a maximum quantifiable plasma concentration C of about 900 ng / mL to about 2100 ng / mL following administration to a subject in need of treatment. max (ng / mL). In other embodiments, the dose achieves a maximum quantifiable plasma concentration of about 1100 ng / mL to about 1900 ng / mL after administration to a subject in need of treatment. In further embodiments, the dose achieves a maximum quantifiable plasma concentration of about 1200 ng / mL to about 1700 ng / mL after administration to a subject in need of treatment. max may be accomplished in a time period of about 1 hour to about 25 hours, about 2 hours to about 10 hours, about 2 hours to about 5 hours, or about 4 hours.

[0113]

[0120] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient. In such an embodiment, the oral dosage form has a C of about 3.0 ng / mL to about 4.0 ng / mL per mg of H3B-6545. max In a further embodiment, the oral dosage form can achieve a C of about 3.0 ng / mL to about 3.4 ng / mL per mg of H3B-6545. max A range can be achieved.

[0114]

[0121] In some embodiments, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable excipient. In some of these embodiments, the dose achieves a half-life of about 9.5 hours to about 14.5 hours, or a half-life of about 10.0 hours to about 11.0 hours after administration to a human subject in need thereof.

[0115]

[0122] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability of about 9,000 ng*hr / mL to about 23,000 ng*hr / mL, or about 12,000 ng*hr / mL to about 19,000 ng*hr / mL, following administration to a human subject in need thereof.

[0116]

[0123] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0-24 hours) of about 30 ng*hr / mL / mg to about 40 ng*hr / mL / mg, or about 30 ng*hr / mL / mg to about 37 ng*hr / mL / mg, measured on a "per mg" basis, after administration to a human subject in need thereof.

[0117]

[0124] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose, following administration to a human subject in need thereof, may achieve a quantifiable AUC bioavailability (0 to infinity time) of about 11,000 ng*hr / mL to about 40,000 ng*hr / mL, about 11,000 ng*hr / mL to about 33,000 ng*hr / mL, or about 12,000 ng*hr / mL to about 40,000 ng*hr / mL.

[0118]

[0125] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0 to infinity) of about 37 ng*hr / mL / mg to about 66 ng*hr / mL / mg, about 37 ng*hr / mL / mg to about 54 ng*hr / mL / mg, or about 38 ng*hr / mL / mg to about 66 ng*hr / mL / mg, measured on a "per mg of H3B-6545" basis, after administration to a human subject in need thereof.

[0119]

[0126] C. An oral dosage form comprising about 450 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient.

[0120]

[0127] In some embodiments, the oral dosage form comprises about 450 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such embodiments, the dose provides a maximum quantifiable plasma concentration C of about 1,000 ng / mL to about 1,600 ng / mL following administration to a subject in need of treatment. max(ng / mL). In other embodiments, the dose achieves a maximum quantifiable plasma concentration of about 1,100 ng / mL to about 1,500 ng / mL after administration to a subject in need of treatment. In further embodiments, the dose achieves a maximum quantifiable plasma concentration of about 1,200 ng / mL to about 1,400 ng / mL after administration to a subject in need of treatment. max can be achieved in a time period of about 3 hours to about 6 hours, or about 4 hours to about 6 hours.

[0121]

[0128] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the oral dosage form has a C of about 2.4 ng / mL to about 3.5 ng / mL per mg of H3B-6545. max In a further embodiment, the oral dosage form can achieve a C of about 3.0 ng / mL to about 3.5 ng / mL per mg of H3B-6545. max In yet a further embodiment, the oral dosage form can achieve a C of about 3.3 ng / mL to about 3.5 ng / mL per mg of H3B-6545. max A range can be achieved.

[0122]

[0129] In some embodiments, the oral dosage form comprises about 450 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In some of these embodiments, the dosage form achieves a half-life of about 8.0 hours to about 11.0 hours, or a half-life of about 9.0 hours to about 10.0 hours after administration to a human subject in need thereof.

[0123]

[0130] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0-24 hours) of about 12,000 ng*hr / mL to about 20,000 ng*hr / mL, about 14,000 ng*hr / mL to about 16,000 ng*hr / mL, or about 12,000 ng*hr / mL to about 17,000 ng*hr / mL after administration to a human subject in need thereof.

[0124]

[0131] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0-24 hours) of about 27 ng*hr / mL / mg to about 43 ng*hr / mL / mg, about 30 ng*hr / mL / mg to about 40 ng*hr / mL / mg, or about 30 ng*hr / mL / mg to about 35 ng*hr / mL / mg, measured on a "per mg of H3B-6545" basis, following administration to a human subject in need thereof.

[0125]

[0132] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dosage form may achieve a quantifiable AUC bioavailability (0 to infinity time) of about 16,000 ng*hr / mL to about 26,000 ng*hr / mL, about 16,000 ng*hr / mL to about 23,000 ng*hr / mL, or about 16,000 ng*hr / mL to about 18,000 ng*hr / mL after administration to a human subject in need thereof.

[0126]

[0133] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharma- ceutically acceptable salt thereof and at least one pharma- ceutically acceptable excipient. In such an embodiment, the dose may achieve a quantifiable AUC bioavailability (0 to infinity) of about 36 ng*hr / mL / mg to about 57 ng*hr / mL / mg, about 38 ng*hr / mL / mg to about 52 ng*hr / mL / mg, or about 35 ng*hr / mL / mg to about 40 ng*hr / mL / mg, measured on a "per mg of H3B-6545" basis, following administration to a human subject in need thereof.

[0127]

[0134] Embodiments may also relate to a method of treating breast cancer (i.e., that is ERα-positive and / or HER2-negative), the method comprising administering to a human subject in need thereof any of the oral dosage forms described herein to obtain the described PK results.

[0128]

[0135] In all embodiments relating to the methods of treatment, the human subject in need thereof may be in a fed or fasted state.

[0129]

[0136] Preclinical trials support dose-dependent inhibition of ERαWT- and ERαmut-dependent transcription and subsequent reduction in cell proliferation. The safety, pharmacokinetics (PK), and pharmacodynamics (PD) of H3B-6545 in women with ER+, HER2-negative breast cancer (BC) will identify a recommended phase 2 dose (RP2D).

[0130]

[0137] Constitutive activating mutations in the ESR1 gene, found in approximately 30% of MBC tumors, are located within the ligand-binding domain and may confer resistance to estrogen deprivation therapy (e.g., aromatase inhibition) and promote resistance to anti-ER therapy. Li S.,et al.Cell Reports,2013;4:1116-1130, Robinson DR,Wu YM,Vats P,et al.Nature genetics,2013;45:1446-1451, Toy W,Shen Y, Won H,et al.Nature genetics.2013;45:1439-1445.

[0131]

[0138] H3B-6545 is an orally available selective estrogen receptor alpha (ERα) covalent antagonist that inhibits both mutant (MUT) and wild-type (WT) activity of estrogen receptor alpha (ERα) by irreversibly engaging a unique cysteine ​​that is not conserved in other nuclear hormone receptors. H3B-6545 potently suppresses ERα function without degrading the receptor.

[0132]

[0139] An embodiment provides a method of treating a patient with H3B-6545 or a pharma- ceutically acceptable salt thereof, comprising identifying whether one or more decreases in mutant allele frequency are observed in the patient's blood, and selecting the patient if said decreases are observed, and administering to the selected patient a therapeutically effective amount of H3B-6545 or a pharma- ceutically acceptable salt thereof. Mutant allele frequency is the relative frequency of a mutant allele (a genetic variant) at a particular locus in a population, expressed as a fraction or percentage.

[0133]

[0140] We report a phase 1 dose escalation study of H3B-6545, a first-in-class highly selective ERα covalent antagonist (SERCA), in women with ER-positive, HER2-negative breast cancer (HR+BC).

[0141] EXAMPLES

[0134]

[0142] Example 1: Capsule formulation

[0135]

[0143] Example 1 reports the formulation of capsules containing various amounts of H3B-6545 drug substance. The capsules used in the subsequent examples were made using this method. Capsules of 25 mg, 50 mg, and 150 mg strengths were formulated according to the following table.

[0136] [Table 1]

[0137] [Table 2]

[0138] [Table 3]

[0139]

[0144] Briefly, the capsules were made by forming an initial mixture of H3B-6545 drug substance, lactose monohydrate, low-substituted hydroxypropylcellulose, microcrystalline cellulose, hydroxypropylcellulose, and colloidal anhydrous silica in a high shear mixer. Magnesium stearate was then mixed with the initial mixture using the high shear mixer. The resulting mixture was then compressed into ribbons using a roller compactor, passed through a screen (sizing), and mixed with magnesium stearate using a tumbler mixer. The final mixture was filled into hypromellose capsules using an encapsulator.

[0140]

[0145] Example 2: Tablet Formation

[0141]

[0146] Example 2 reports the formation of tablets containing H3B-6545 drug substance. The tablets discussed in the following examples were made using this method. H3B-6545 fill-coated tablets containing 150 mg of H3B-6545 drug substance as free base were made according to the following formulation:

[0142] [Table 4]

[0143] [Table 5]

[0144]

[0147] Tablets were formed in the following manner:

[0145]

[0148] Lactose monohydrate, low substituted hydroxypropyl cellulose, hypromellose, and colloidal silicon dioxide were added to a wet high shear granulator and mixed to form a first mixture. The H3B-6545 drug substance and the first mixture were then added to a convection mixer to form a second mixture.

[0146]

[0149] The second mixture was continuously fed into a wet high shear granulator and granulated with purified water pumped in to form wet granules. The wet granules were continuously dried at elevated temperature using a fluid bed dryer until the loss on drying of the dry granules reached 1.5% or less.

[0147]

[0150] The dried granules were then milled using a screen mill. The milled granules, microcrystalline cellulose, and magnesium stearate were then placed in a convection mixer and mixed to form a mixed granule that was compressed into a core tablet. The coating suspension (prepared by suspending the coating agent in purified water) was then sprayed onto the core tablet using a pan coating machine.

[0148]

[0151] Blood samples were collected from each patient at scheduled time points and centrifuged. The plasma portion of each sample was then transferred to a K2EDTA tube, which was then transported to a biochemical analysis laboratory for concentration measurement using LC-MS / MS. The concentration data was then analyzed using Phoenix® WinNonlin software to obtain PK parameters. The obtained PK parameters were further summarized using the analysis software.

[0149]

[0152] Blood samples (approximately 5 mL each) were collected from human patients at various time points pre- and post-dose. Plasma concentrations of H3B-6545 were determined using a validated high performance liquid chromatography / tandem mass spectrometry (LC-MS / MS) method. The lower limit of quantification was 0.100 ng / mL.

[0150]

[0153] The PK analysis set included subjects who received the H3B-6545 drug product in capsules or tablets and had sufficient evaluable plasma concentration data to obtain at least one primary PK parameter for each treatment. Plasma concentrations of H3B-6545 (as the free base) were tableted and summarized at each nominal time point using descriptive statistics (number of subjects, arithmetic mean, coefficient of variation [CV%], geometric mean, median, minimum, and maximum values ​​shown with standard deviation [SD]) as appropriate. Individual and mean (±SD) plasma concentration-time profiles were obtained for each treatment.

[0151]

[0154] Pharmacokinetic parameters of H3B-6545 were calculated using non-compartmental methods using the plasma data and an appropriate model for extravascular administration. PK parameters were calculated using the area under the curve to the last quantifiable concentration (AUC 0-t ), maximum concentration (C max ), and the time when the maximum concentration occurs (t max If data permit, the area under the plasma concentration-time curve is extrapolated to infinity (AUC 0-inf ), terminal elimination half-life (t 1 / 2), apparent total body clearance (CL / F), total total body clearance after oral administration at steady state (CL ss / F), apparent volume of distribution during the terminal phase (Vz / F), apparent steady-state volume of distribution (Vss), C max (RC max ) Accumulation ratio and AUC 0-24 The accumulation ratio of (RAUC) was also obtained.

[0152]

[0155] Example 3: First Interim Analysis of Capsule Pharmacokinetics (PK) in Patients in the Fasting State

[0153]

[0156] Example 3 reports the clinical and genomic analysis of a single study. The study population in Example 3 was as follows: Women aged 18 years or older with locally advanced or metastatic ER+HER2-BC Progression on at least one hormonal therapy and at least one additional therapy / regimen, no maximum prior therapy ECOG performance status of 0 or 1 Adequate bone marrow and organ function Remove if present with bone only disease or inflammatory BC.

[0154]

[0157] Methods were used to analyze whether H3B-6545 inactivates both wild-type and mutant ERα by targeting cysteine ​​530 and forcing it to adopt a unique antagonist conformation. Methods: Women with locally advanced or metastatic HR+BC are treated (tx) with H3B-6545 drug product administered orally in capsules once daily for 28-day cycles after progression on at least one hormonal treatment and at least one additional treatment / regimen. Dose escalation uses a 3+3 design with the option to backfill previously excluded doses, allowing for dose escalation between patients. This example investigated the safety, pharmacokinetics, and pharmacodynamics of H3B-6545 in women with HR+BC to identify recommended subsequent doses for testing.

[0155]

[0158] RESULTS:As of December 10, 2018, 32 patients had been treated with H3B-6545 drug product at doses ranging from 100 to 450 mg / day, 97% had received prior treatment with a CDK4 / 6 inhibitor, and 56% had received ≥3 lines of prior anticancer therapy. No dose-limiting toxicities were observed, and only one grade 3 treatment-related adverse event (TRAE) was observed (lymphocyte count decrease).

[0156]

[0159] The most common (≥10%) TRAEs included asymptomatic sinus bradycardia, diarrhea, nausea, fatigue, anemia, decreased appetite, and hot flashes. H3B-6545 was effective in treating ADHD with a t max Absorption was rapid at 1 mg / kg / day. Plasma concentrations increased at doses of 100-450 mg and were comparable in C1D1 and C1D15. Consistent with H3B-6545's mechanism of action and preclinical data, H3B-6545 inhibits ER target gene expression and shows a 50% decrease in Ki67 at all dose levels after treatment. ESR1 (60%) and PIK3CA (34%) mutations were detected in plasma at baseline, and changes in mutant allele frequency correlated with response to treatment. Stable disease was observed in 15 patients (47%), with 34% of patients completing at least 6 months of treatment. Partial responses (PRs) were observed in three patients; one patient (mutant) had received two lines of prior therapy, two patients (one mutant and one wild-type) had received more than five lines of prior therapy including fulvestrant and capecitabine, and all three patients had received prior CDK4 / 6 inhibitors.

[0157]

[0160] The H3B-6545 drug product was well tolerated up to the 450 mg dose level, where early signs of single-agent antitumor activity were seen in the historical CDK4 / 6 setting. Dose escalation continued in patients with advanced HER2-negative breast cancer.

[0158]

[0161] Drug Administration and Dosage: H3B-6545 was administered orally once daily for 28-day cycles. Dose escalation was performed using a standard 3+3 study, with dose cohorts of 100, 200, 300, 450, and 600 mg QD. Patients were able to backfill previously excluded doses. Dose escalation among patients was permitted after completion of cycle 3.

[0159]

[0162] Results (as of April 20, 2019)

[0160]

[0163] Patient Population: Forty-six patients were treated with H3B-6545 at doses ranging from 100 to 600 mg QD. Forty-two (91.3%) patients had received prior CDK4 / 6 inhibitors and 32 (69.6%) had received prior fulvestrant. Thirty (65.2%) patients had received ≥3 lines of prior anticancer therapy in the advanced / metastatic setting.

[0161]

[0164] Demographic and baseline characteristics are shown in Table 6.

[0162] [Table 6] TIFF2025038130000011.tif58149

[0163]

[0165] Safety: Two dose-limiting toxicities (DLTs), grade 3 fatigue and grade 3 cardiac urticaria, occurred in the 600 mg QD cohort. The most common (≥10%) treatment-related adverse events (TRAEs) included sinus bradycardia, nausea, fatigue, anemia, diarrhea, and aspartate aminotransferase (Table 2). Six grade 3 TRAEs were observed: syncope and electrocardiogram QT prolongation at 450 mg, anemia, increased aspartate aminotransferase, fatigue, and rash urticaria at 600 mg. There were no grade 4 or higher TRAEs. The most common TRAE, sinus bradycardia, did not appear to be dose or concentration dependent and did not require dose reduction, interruption, or discontinuation.

[0164]

[0166] TRAEs are summarized in Table 7.

[0165] [Table 7]

[0166]

[0167] Pharmacokinetics: t max was 2 to 4 hours. Plasma concentrations increased with dose (100-600 mg) (Figure 1). Plasma concentrations were comparable between C1D1 and C1D15.

[0167]

[0168] Pharmacodynamics: ESR1 (55%) and PIK3CA (39%) mutations were detected in baseline plasma. Changes in mutant allele frequency correlate with clinical response.

[0168]

[0169] Patient Case Studies:

[0169]

[0170] One, a 50-year-old woman originally diagnosed with ER+HER2 breast cancer in 2006, had received prior therapy including letrozole / palbociclib, entinostat / exemestane, capecitabine, eribulin, and carboplatin / gemcitabine in the metastatic setting. ESR1 Y537S and PI3KCA E545K mutations were detected in baseline tumor and plasma. At the time of enrollment, ECOG performance status was 1, and disease sites included liver, bone, pleural effusion, pelvic ascites, and subcutaneous nodules. H3B-6545 was initiated in June 2018 at a dose of 450 mg QD.

[0170]

[0171] The reduction in the sum of the diameters of all target lesions from baseline to C3D1 and C7D1 was approximately -27.8% and -35.6%, respectively. After four cycles, the patient achieved a partial response, which was maintained on treatment at C11.

[0171]

[0172] Preliminary Activity: Stable disease was observed in 17 (37.0%) patients. At least 6 months of treatment was completed by 14 (30.4%) patients. Confirmed partial responses were observed in three (6.5%) patients.

[0172]

[0173] Further information on preliminary activity is shown in Figure 2. Figure 3 shows the percent change in the sum of the diameters of the target lesions.

[0173]

[0174] Table 8 shows the tumor response and progression-free survival.

[0174] [Table 8] TIFF2025038130000014.tif62149

[0175]

[0175] Conclusion Safety / Tolerability: Most TRAEs were grade 1 or 2, were manageable, and were tolerated up to, and including, 450 mg QD. Pharmacodynamics: Changes in variant allele frequency show a correlation to clinical response. Preliminary Activity: Encouraging signals of antitumor activity were observed in the heavily pretreated, post-CDK4 / 6i setting, including 3 confirmed partial responses and mPFS=7.2 months. Phase 2: Must begin after determination of RP2D.

[0176]

[0176] Example 4: Second Interim Analysis

[0177]

[0177] The clinical trial of Example 3 was continued, and PK evaluation (of this Example 4) was performed at a later date based on a larger patient population. Table 1 summary of the pharmacokinetic (PK) analysis of capsules in fasting patients and Table 2 show the PK parameters and profile of human patients administered capsules of H3B-6545 drug product prepared as in Example 1. Patients received doses of 100 mg, 200 mg, 300 mg, 450 mg, or 600 mg total equivalent of Formula I, as specified in the table. Table 1 reflects the PK values ​​for patients measured on day 1 of the first cycle. Table 2 reflects the PK values ​​for patients measured on day 15 of the first cycle.

[0178]

[0178] PK parameters were calculated after administration to human subjects in need thereof, where the human subjects were in the fasting state. QD means that the dose was administered "quaque die" or only once a day. The parameter n is the number of human subjects in need thereof that received each dose.

[0179] Tables 9 and 11 show the T max and T lag The geometric means and geometric mean coefficients of variation for all plasma PK parameters, except for T, are calculated from human subjects (n) in need thereof. max , and T lag The values ​​include the minimum and maximum values ​​as well as the median values ​​calculated among human subjects in need thereof. Tables 10 and 12 show the AUC 0-t ,AU0C 0-24 , AUC0 0-inf , and C max PK parameters are normalized to plasma PK parameters per mg of active pharmaceutical ingredient (API).

[0180] [Table 9]

[0181] [Table 10]

[0182] [Table 11]

[0183] [Table 12]

[0184]

[0180] Example 5: Pharmacokinetics (PK) of 450 mg Capsules in Fasting and Fed Patients

[0185] Table 3 details the PK parameters and profiles of human patients administered capsules prepared as in Example 1. Patients received a total equivalent dose of 450 mg of Formula I as specified in Table 13. Table 14 shows the AUC 0-t , AUC 0-24 , AUC 0-inf , and C max PK parameters are normalized to plasma PK parameters per mg of active pharmaceutical ingredient (API).

[0186] [Table 13]

[0187] [Table 14]

[0188]

[0182] Example 6: Pharmacokinetics (PK) of tablets in humans

[0189] The capsules described in Example 1 and the tablets described in Example 2 were tested in healthy human subjects. All doses were well tolerated. Similar exposures (C max and AUC) and T max was noted between capsules and tablets as shown in Table 15.

[0190] [Table 15]

[0191]

[0184] The relative bioavailability of capsule vs. tablet formulations of H3B-654 is shown in Table 16.

[0192] [Table 16]

[0193]

[0185] Example 7 - Genomics Testing

[0194]

[0186] Due to the lack of effective treatments for endocrine-resistant metastatic breast cancer (MBC), the inventors developed H3B-6545, a novel selective Era covalent antagonist that can irreversibly inactivate both wild-type and mutant Era.

[0195]

[0187] The objectives of this study are: 1) to characterize the hotspot mutation profile in heavily pretreated MBC and correlate ESR1, PIK3CA, and AKT1 mutations in plasma vs tumor tissue, 2) to determine whether mutations in ESR1 or PIK3CA predict response to H3B-6545, and 3) to evaluate whether long-term tracking of ctDNA correlates with response to H3B-6545.

[0196]

[0188] Method

[0197] Fresh plasma samples were collected at baseline (pre-dose), cycle 1, day 15 (C1D15), C2D1, C3D1 and every 8 weeks thereafter, with a final sample collection at disease progression. At baseline, hotspot mutations in ESR1, PIK3CA, and AKT1 were assessed using beaming digital PCR. Patient-specific ctDNA mutations were then monitored by ddPCR. Baseline tumor biopsies were subjected to a targeted next-generation sequencing (NGS) panel to identify hotspot mutations.

[0198]

[0190] Results

[0199] In 77% (30 / 39) of patients, mutations were detected at baseline by the Beaming assay; of these, 21 / 39, 16 / 39, and 3 / 39 had mutations in ESR1, PIK3CA, and AKT1, respectively. 20% (9 / 39) of patients showed co-mutations in ESR1 and PIK3CA. In 60% (9 / 15) of patients, DNA mutations identified by the plasma Beaming assay were also detected by tumor biopsy, where DNA mutations found in tissue were also detected in plasma in 86% (12 / 14) of cases. Serial ctDNA monitoring revealed that in patients with confirmed partial responses (3 / 3), ctDNA levels were undetectable by C2D1. In contrast, ctDNA levels increased from baseline in 3 / 4 of patients with progressive disease. Testing of ctDNA ratios (day 15 / baseline and day 30 / baseline) and correlation of PIK3CA and ESR1 mutations with response to H3B-6545 are shown.

[0200] Conclusions for Example 7

[0201]

[0193] ctDNA is a reliable sample type for evaluating ESR1, PIK3CA, and AKT1 mutations in MBC, overcoming the challenges of obtaining biopsies in the metastatic setting. Furthermore, ctDNA kinetics is believed to provide a useful tool for monitoring the efficacy of H3B-6545.

[0202] Drug Administration and Dosage: H3B-6545 administered once daily (PO QD) on a 28-day cycle Dose escalation using a standard 3+3 study design with dose cohorts of 100, 200, 300, 450, and 600 mg / day

[0203] Liquid biopsy assay: 10 ml of Strek plasma was collected at the site and then processed to plasma in Sysmex Inostics. cfDNA was isolated from 2 ml of plasma for beaming and 3 ml for Biorad ddPCR.

[0204]

[0196] Tumor next-generation sequencing (NGS) analysis: After DNA from 10x5 μm slides was isolated using the Recoverall Kit, 10 ng of DNA was used as input for the Oncomine comprehensive assay.

[0205]

[0197] Study baseline characteristics are shown in Table 17.

[0206] [Table 17]

[0207]

[0198] Figure 4 shows an overview of the liquid and tissue biopsy sample sets.

[0208]

[0199] Figures 5A-5E show the baseline genomics profile of the patients in this example. AA = amino acid. *AKT1 mutations found in 3 / 49 patients (data not shown).

[0209]

[0200] Figures 5A-5E show BEAMing assays: A. Baseline ESR1 and PIK3CA mutation status of patients, B and C. Clonality of ESR1 and PIK3CA mutations, D and E. Amino acid distribution of PIK3CA and ESR1 mutations.

[0210]

[0201] Figure 5F shows the OncoPrint of mutations found in baseline tissue biopsies as determined by the Oncomine comprehensive panel.

[0211]

[0202] Figures 6A and 6B show the concordance of mutations found in tissue and liquid biopsies in Example 2. Figure 6A shows the allele frequency distribution and concordance of tissue and liquid biopsy mutations. Figure 6B is a table summarizing the concordance of tissue and liquid biopsy mutations.

[0212]

[0203] Figures 7A-7C show ctDNA kinetics correlating with overall best response to H3B-6545. AF = allele frequency, C = cycle, D = day, PD = progressive disease, PR = partial response, SD = stable disease. Dotted lines represent ddPCR assay LOD. Figure 7A. AF baseline / C2D1 ratio for each patient colored separately. One patient may have multiple mutations. Figures 7B and 7C. Exemplary ctDNA kinetics in patients with progressive disease (B) vs. partial response (C).

[0213]

[0204] Patient Case Study - This is the same patient case study reported in Example 3. A 50-year-old woman was initially diagnosed with ER+HER2-BC in 2006. Prior therapy in the metastatic setting included letrozole / palbociclib, entinostat / exemestane, capecitabine, eribulin, and carboplatin / gemcitabine. At the time of enrollment, ECOG performance status was 1 and disease sites included liver, bone, pleural effusion, pelvic ascites, and subcutaneous nodules. ESR1 Y537S and PI3KCA E545K mutations were detected in baseline tumors and plasma. H3B-6545 was initiated in June 2018 at a dose of 450 mg PO QD. The reduction in the sum of the diameters of all target lesions from baseline to C3D1 and C7D1 was -27.8% and -35.6%, respectively. After four cycles, the patient achieved a partial response that was maintained during treatment with C11.

[0214]

[0205] Conclusion In late-line ER+ MBC, PIK3CA and ESR1 mutations were detected in 39% and 55% of baseline plasma, respectively. Concordance to tissue plasma mutation levels was 80%. Concordance of liquid biopsy to tissue mutation levels was 51%. ctDNA levels were undetectable by C2D1 in patients with confirmed partial responses. ctDNA kinetics may be a useful tool for monitoring the efficacy of H3B-6545 and for baseline genomics profiling.

[0215]

[0206] All publications and patent documents cited in this specification are incorporated herein by reference as if such publications or documents were specifically and individually indicated to be incorporated herein by reference. Citation of publications and patent documents is not intended as an admission that any is a relevant art, nor does it constitute an admission as to the contents or date thereof. The invention has now been described by specification, and those skilled in the art will recognize that the invention may be embodied in various embodiments, and that the foregoing description and examples are intended to be illustrative and not limiting on the scope of the following claims.

[0216]

[0207] The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. The foregoing embodiments are therefore intended in all respects to be illustrative rather than limiting of the invention described herein. The scope of the invention is, therefore, indicated by the appended claims, rather than the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Claims

1. i) a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; 1. An oral dosage form comprising: The formula I has the structure 【Chemistry 1】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide represented by the formula: When the oral dosage form is orally administered once daily to a human subject, it has a mean C of about 1 ng / mL to about 4 ng / mL, measured for each mg of Formula I in the dosage form. ｍａｘ The oral dosage form is formulated to achieve the following:

2. Said average C ｍａｘ is from about 2 ng / mL to about 4 ng / mL for each mg of Formula I in the dosage form.

3. Said average C ｍａｘ is from about 3 ng / mL to about 4 ng / mL for each mg of Formula I in the dosage form.

4. Said average C ｍａｘ for each mg of Formula I in said dosage form: From 80% to 125% of 3 ng / mL, 2. The oral dosage form of claim 1, wherein the concentration is in the range of 80% to 125% of 3.5 ng / mL.

5. About 2 hours to about 7 hours, the average C ｍａｘ The average of t ｍａｘ The oral dosage form according to any one of claims 1 to 4, which is formulated to achieve:

6. About 3 hours to about 7 hours, the average C ｍａｘ The average of t ｍａｘ 6. The oral dosage form of claim 5, formulated to achieve:

7. About 3.5 hours to about 4.5 hours, the average C ｍａｘ The average of t ｍａｘ 7. The oral dosage form of claim 6, formulated to achieve:

8. About 5.5 hours to about 6.5 hours, ｍａｘ The average of t ｍａｘ 7. The oral dosage form of claim 6, formulated to achieve:

9. 7. The oral dosage form of any one of claims 1 to 6, comprising a total equivalent amount of formula I of about 100 mg to about 600 mg.

10. 7. The oral dosage form of any one of claims 1 to 6, comprising a total equivalent amount of about 450 mg of formula I.

11. i) a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; 1. An oral dosage form comprising: The formula I has the structure 【Chemistry 2】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, When the oral dosage form is orally administered once daily to a human subject, the mean AUC of about 16 h*ng / mL to about 44 h*ng / mL for each mg of Formula I in the dosage form. ０－２４ The oral dosage form is formulated to achieve the following:

12. The average AUC ０－２４ is from about 27 h*ng / mL to about 44 h*ng / mL for each mg of Formula I in the dosage form.

13. The average AUC ０－２４ for each mg of Formula I in said dosage form: From 80% to 125% of 30h*ng / mL, 12. The oral dosage form of claim 11, wherein the NADH concentration is in the range of 80% to 125% of 44h*ng / mL.

14. 12. The oral dosage form of claim 11, comprising from about 100 mg to about 600 mg total equivalents of Formula I.

15. 12. The oral dosage form of claim 11, comprising about 450 mg total equivalent of formula I.

16. i) a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; 1. An oral dosage form comprising: The formula I has the structure 【Chemistry 3】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, When the oral dosage form is administered once daily to a human subject, the average t １／２ The oral dosage form is formulated to achieve the following:

17. The average t １／２ is from about 8 hours to about 12 hours.

18. The average t １／２ 18. The oral dosage form of claim 17, wherein the time from about 9 hours to about 11 hours.

19. 17. The oral dosage form of claim 16, comprising from about 100 mg to about 600 mg total equivalents of formula I.

20. 17. The oral dosage form of claim 16, comprising about 450 mg total equivalent of formula I.

21. i) a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; 1. An oral dosage form comprising: The formula I has the structure 【Chemistry 4】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, When the oral dosage form is administered once daily to a human subject, the mean AUC of about 21 h*ng / mL to about 67 h*ng / mL for each mg of Formula I in the dosage form. ０－ｉｎｆ The oral dosage form is formulated to achieve the following:

22. Average AUC ０－ｉｎｆ is from about 29 h*ng / mL to about 67 h*ng / mL for each mg of Formula I in the dosage form.

23. The average AUC ０－ｉｎｆ for each mg of Formula I in said dosage form: From 80% to 125% of 36h*ng / mL, 22. The oral dosage form of claim 21, in the range of 80% to 125% of 57 h*ng / mL.

24. 22. The oral dosage form of claim 21 comprising from about 100 mg to about 600 mg total equivalents of formula I.

25. 22. The oral dosage form of claim 21, comprising about 450 mg total equivalent of formula I.

26. i) a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; 1. An oral dosage form comprising: The formula I has the structure 【Chemistry 5】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, When the oral dosage form is administered once daily to a human subject, the mean AUC of about 16 h*ng / mL to about 41 h*ng / mL for each mg of Formula I in the dosage form. ０－ｔ The oral dosage form is formulated to achieve the following:

27. The average AUC ０－ｔ for each mg of Formula I in said dosage form: From 80% to 125% of 27h*ng / mL, 27. The oral dosage form of claim 26, in the range of 80% to 125% of 36h*ng / mL.

28. 27. The oral dosage form of claim 26, comprising from about 100 mg to about 600 mg total equivalents of formula I.

29. 27. The oral dosage form of claim 26, comprising about 450 mg total equivalent of formula I.

30. i) an internal phase comprising a composition of Formula I or a pharma- ceutically acceptable salt, lactose monohydrate, low-substituted hydroxypropylcellulose, microcrystalline cellulose, hydroxypropylcellulose, colloidal anhydrous silica, and magnesium stearate; and ii) An outer phase comprising magnesium stearate 30. The oral dosage form of claim 1, 11, 16, 21, or 26, which is a capsule comprising:

31. 31. The oral dosage form of claim 30, wherein the capsule is a hypromellose capsule.

32. 32. The oral dosage form of claim 31 comprising a mono-HCl salt form of formula I.

33. i) an internal phase comprising a composition of formula I or a pharma-ceutically acceptable salt thereof, lactose monohydrate, low-substituted hydroxypropylcellulose, hypromellose, colloidal silicon dioxide, and purified water; ii) an outer phase comprising microcrystalline cellulose and magnesium stearate; and iii) a film coating comprising hypromellose, talc, titanium dioxide, propylene glycol, iron(III) oxide, and purified water.

27. The oral dosage form of any one of claims 1, 11, 16, 21, and 26, which is a tablet comprising:

34. 34. The oral dosage form of claim 33 comprising a mono-HCl salt form of formula I.

35. 1. A method of treating cancer in a human subject, comprising: i) a therapeutically effective amount of a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; administering to the subject an oral dosage form comprising The formula I has the structure 【Chemistry 6】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, said therapeutically effective amount being a single daily dose ranging from about 100 mg to 600 mg; The oral dosage form has a mean C of about 1 ng / mL to about 4 ng / mL in the subject's blood plasma for each mg of Formula I in the dosage form. ｍａｘ The method comprising:

36. Said average C ｍａｘ is from about 2 ng / mL to about 4 ng / mL for each mg of Formula I in the dosage form.

37. Said average C ｍａｘ is from about 3 ng / mL to about 4 ng / mL for each mg of Formula I in the dosage form.

38. Said average C ｍａｘ for each mg of Formula I in said dosage form: From 80% to 125% of 3 ng / mL, The method of claim 35, wherein the concentration is in the range of 80% to 125% of 3.5 ng / mL.

39. The dosage form has a mean C ｍａｘ The average of t ｍａｘ The method according to any one of claims 35 to 38, comprising:

40. The dosage form has a mean C ｍａｘ The average of t ｍａｘ 40. The method of claim 39, having the following structure:

41. The dosage form has a mean C ｍａｘ The average of t ｍａｘ 41. The method of claim 40, having the following structure:

42. The dosage form has a mean C ｍａｘ The average of t ｍａｘ 41. The method of claim 40, having the following structure:

43. 1. A method of treating cancer in a human subject, comprising: i) a therapeutically effective amount of a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; administering to the subject an oral dosage form comprising The formula I has the structure 【Chemistry 7】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, The oral dosage form has a mean AUC of about 16 h*ng / mL to about 44 h*ng / mL for each mg of Formula I in the dosage form. ０－２４ The method comprising:

44. The average AUC ０－２４ is from about 27 h*ng / mL to about 44 h*ng / mL for each mg of Formula I in the dosage form.

45. The average AUC ０－２４ for each mg of Formula I in said dosage form: From 80% to 125% of 30h*ng / mL, 44. The method of claim 43, wherein the concentration of 44h*ng / mL is in the range of 80% to 125%.

46. 1. A method of treating cancer in a human subject, comprising: i) a therapeutically effective amount of a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; administering to the subject an oral dosage form comprising The compound of formula I has the structure 【Chemistry 8】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, The oral dosage form has a mean t １／２ The method comprising:

47. The average t １／２ is from about 8 hours to about 12 hours.

48. The average t １／２ is from about 9 hours to about 11 hours.

49. 1. A method of treating cancer in a human subject, comprising: i) a therapeutically effective amount of a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; administering to the subject an oral dosage form comprising The formula I has the structure 【Chemistry 9】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, The oral dosage form has a mean AUC of about 21 h*ng / mL to about 67 h*ng / mL for each mg of Formula I in the dosage form. ０－ｉｎｆ The method comprising:

50. Average AUC ０－ｉｎｆ is from about 29 h*ng / mL to about 67 h*ng / mL for each mg of Formula I in the dosage form.

51. The average AUC ０－ｉｎｆ for each mg of Formula I in said dosage form: From 80% to 125% of 36h*ng / mL, 57h*ng / mL.

52. 1. A method of treating cancer in a human subject, comprising: i) a therapeutically effective amount of a compound of formula I or a pharma- ceutically acceptable salt thereof; ii) at least one pharma- ceutically acceptable excipient; administering to the subject an oral dosage form comprising The formula I has the structure 【Chemistry 10】 (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazol-5-yl)-2-phenylbut-1-enyl]pyridin-2-yl]oxyethylamino]but-2-enamide, The oral dosage form has a mean AUC of about 16 h*ng / mL to about 41 h*ng / mL for each mg of Formula I in the dosage form. ０－ｔ The method comprising:

53. The average AUC ０－ｔ for each mg of Formula I in said dosage form: From 80% to 125% of 27h*ng / mL, 53. The method of claim 52, wherein the concentration is in the range of 80% to 125% of 36h*ng / mL.

54. 53. The method of any one of claims 35, 43, 46, 49 and 52, wherein the dosage form comprises a total equivalent amount of formula I of about 100 mg to about 600 mg.

55. 55. The method of claim 54, wherein the dosage form comprises a total equivalent of from about 200 mg to about 600 mg of Formula I.

56. 56. The method of claim 55, wherein the dosage form comprises a total equivalent of from about 300 mg to about 600 mg of Formula I.

57. 57. The method of claim 56, wherein the dosage form comprises the equivalent of about 450 mg total of formula I.

58. The oral dosage form comprises: i) an internal phase comprising a composition of Formula I or a pharma- ceutically acceptable salt, lactose monohydrate, low-substituted hydroxypropylcellulose, microcrystalline cellulose, hydroxypropylcellulose, colloidal anhydrous silica, and magnesium stearate; and ii) An outer phase comprising magnesium stearate 42. The method of any one of claims 24, 32, 35, 38 and 41, wherein the capsule comprises

59. 48. The method of claim 47, wherein the capsule is a hypromellose capsule.

60. 49. The method of claim 48 comprising the mono-HCl salt form of formula I.

61. The oral dosage form comprises: i) an internal phase comprising a composition of Formula I or a pharma-ceutically acceptable salt thereof, lactose monohydrate, low-substituted hydroxypropylcellulose, hypromellose, colloidal silicon dioxide, and purified water; ii) an external phase comprising microcrystalline cellulose and magnesium stearate; and iii) a film coating comprising hypromellose, talc, titanium dioxide, propylene glycol, iron (III) oxide, and purified water.

42. The method of any one of claims 24, 32, 35, 38 and 41, wherein the tablet comprises

62. 62. The method of claim 61 comprising the mono-HCl salt form of formula I.

63. 53. The method of any one of claims 35, 43, 46, 49 and 52, wherein the cancer is breast cancer.

64. The method of claim 61, wherein the breast cancer is an ERα-positive breast cancer.

65. 65. The method of claim 64, wherein the breast cancer expresses wild-type ERa.

66. 65. The method of claim 64, wherein the breast cancer expresses a mutant ERa.

67. 53. The method of any one of claims 35, 43, 46, 49 and 52, wherein the oral dosage form is administered once daily.

68. 58. The method of claim 57, wherein the oral dosage form is administered once daily.

69. 53. The method of any one of claims 35, 43, 46, 49 and 52, wherein the oral dosage form is administered to a human in a fasted state.

70. 53. The method of any one of claims 35, 43, 46, 49 and 52, wherein the oral dosage form is administered to a human in a fed state.

Images