A method of treating cancer using oral estrogen receptor alpha inhibitors.

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 of related applications

[0001]

[0001] This application claims the interests of U.S. Provisional Patent Application No. 62 / 852,751, filed on 24 May 2019, and U.S. Provisional Patent Application No. 62 / 853,498, filed on 28 May 2019. These applications are incorporated herein by reference in their entirety. Background of the Invention

[0002]

[0002] Alterations in estrogen receptor (ER) signaling are known to play a crucial 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 the cell 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, mutations occur within different ERs to develop, where these mutations confer upon cancer cells the ability to activate the ER signaling pathway independently of the ligand. ERα, a specific ER known to be associated with breast cancer tumorigenesis, is a hormone-regulating 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 regulatory 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 is common in clinical practice and is associated with several mechanisms. 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 at the forefront of treatment 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 specific diseases. Endocrine therapy targeting ERα has been shown to be ineffective as a long-term treatment method for breast cancer because 20% to 40% of all breast cancers expressing ERα acquire mutations after long-term endocrine therapy, and these mutations cause the receptor to become unresponsive to conventional endocrine therapy (Robinson, DR, Nat. Genet. 2013; 45: 1446-1451). For this reason, there is a need in the field of chemotherapy to develop novel therapeutic methods that can effectively target mutated ERα isoforms that make cancer resistant to endocrine therapy. While pursuing the development of novel therapeutic methods for endocrine therapy-resistant breast cancer, a novel class of ERα inhibitors called Selective Estrogen Receptor Covalent Antagonists (SERCAs) was discovered. SERCAs inactivate 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 identified during this period is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, which is represented by the following formula I: [ka] This is shown by [the following].

[0005]

[0005] The free base form of (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide is sometimes 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, thereby causing the receptor to induce a specific structure 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 exhibit potent chemotherapeutic properties in various breast cancer cell lines and in 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 has not yet been determined how H3B-6545 should be administered to human breast cancer patients in need of treatment. Therefore, there is a need to provide formulations and administration regimens of the inhibitor that will enable H3B-6545 to be administered more effectively and safely to human subjects in need. Characterizing the pharmacokinetic (PK) results and subsequent PK profile will enable its general use in human breast cancer treatment methods. Brief Overview

[0007]

[0007] Embodiments may also provide an oral dosage form comprising, for example, a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, where formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, and when administered once daily to human subjects, the oral dosage form, when measured for each mg of formula I in the dosage form, yields an average C of approximately 1 ng / mL to approximately 4 ng / mL. max It is formulated to achieve the mean C max The concentration of C is approximately 2 ng / mL to approximately 4 ng / mL for each mg of formula I in the dosage form. In some embodiments, the average C max The concentration of the active ingredient in the dosage form is approximately 3 ng / mL to approximately 4 ng / mL for each mg of formula I. In some embodiments, the average C max The concentration of each mg of formula I in the aforementioned dosage form is in the range of 80% to 125% of 3 ng / mL and 80% to 125% of 3.5 ng / mL.

[0008]

[0008] In a further embodiment, the dosage form is used to reach the average C in about 2 hours to about 7 hours. max The average t maxformulated to achieve. In a further embodiment, the dosage form provides said average C in about 3 hours to about 7 hours, the average max t max is formulated to achieve. In a further embodiment, the dosage form provides said average C in about 3.5 hours to about 4.5 hours, the average max t max is formulated to achieve. In a further embodiment, the dosage form provides said average C in about 5.5 hours to about 6.5 hours, the average max t max is formulated to achieve.

[0009]

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

[0010]

[0010] A further embodiment provides an oral dosage form comprising the compound represented by Formula I or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically 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, and wherein when orally administered once daily to a human subject, the oral dosage form provides an average AUC of from about 16 h*ng / mL to about 44 h*ng / mL per mg of Compound of Formula I in said dosage form 0-24 formulated to achieve. In some embodiments, the average AUC 0-24 is from about 27 h*ng / mL to about 44 h*ng / mL per mg of Compound of Formula I in said dosage form. In some embodiments, the average AUC 0-24 ranges from 80% to 125% of 30 h*ng / mL to 80% to 125% of 44 h*ng / mL per mg of Compound of Formula I in said dosage form. In some embodiments, the dosage form comprises from about 100 mg to about 600 mg total equivalent of Compound of Formula I. In some embodiments, the dosage form comprises about 450 mg total equivalent of Compound of Formula I.

[0011]

[0011] Further embodiments provide an oral dosage form comprising a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, wherein the compound represented by formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, and the oral dosage form, when administered orally once daily to human subjects, provides an average t of formula I in the dosage form for about 8 to about 22 hours. 1 / 2 It is formulated to achieve this. In a further embodiment, average t 1 / 2 This is approximately 8 to 12 hours. In further embodiments, the average t 1 / 2 The incubation period is approximately 9 to 11 hours. In some embodiments, the dosage form contains a total equivalent of approximately 100 mg to 600 mg of formula I. In some embodiments, the dosage form contains a total equivalent of approximately 450 mg of formula I.

[0012]

[0012] Further embodiments provide an oral dosage form comprising a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, wherein the compound represented by formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, and the oral dosage form, when administered orally once daily to human subjects, has an average 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 It is formulated to achieve the mean AUC. In some embodiments, 0-inf The average AUC is approximately 29 h*ng / mL to approximately 67 h*ng / mL for each mg of formula I in the dosage form. In some embodiments, the average AUC 0-infThe amount of each mg of formula I in the dosage form ranges from 80% to 125% of 36 h*ng / mL to 80% to 125% of 57 h*ng / mL. 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] Further embodiments may include dosage forms described herein that have means for achieving the pharmacokinetic values ​​described herein.

[0014]

[0014] Further embodiments provide an oral dosage form comprising a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically 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-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, and the oral dosage form has an average AUC of about 16 h*ng / mL to about 41 h*ng / mL for each mg of formula I in the dosage form when administered orally once daily to human subjects. 0-t It is formulated to achieve the mean AUC. In a further embodiment, the mean AUC 0-t The amount of each mg of formula I in the dosage form ranges from 80% to 125% of 27 h*ng / mL to 80% to 125% of 36 h*ng / mL. 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]

[0015] In further embodiments, the previously reported dosage forms may be capsules comprising an internal phase comprising formula I or a pharmaceutically acceptable salt, lactose monohydrate, low-substituted hydroxypropyl cellulose, microcrystalline cellulose, hydroxypropyl cellulose, 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 a monoHCl salt form of formula I.

[0016]

[0016] In some previously reported embodiments, the oral dosage form is a tablet comprising an internal phase comprising formula I or a pharmaceutically acceptable salt, lactose monohydrate, low-substituted hydroxypropyl cellulose, hypromellose, colloidal silicon dioxide, and purified water; an external 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 a monoHCl salt form of formula I.

[0017]

[0017] Further embodiments provide a method for treating cancer in a human subject, comprising the step of administering to the subject an oral dosage form comprising a therapeutically effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, wherein formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, the therapeutically effective amount is a single daily dose in the range of about 100 mg to 600 mg, and the oral dosage form comprises an average 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 It has the average C max The concentration of C is approximately 2 ng / mL to approximately 4 ng / mL for each mg of formula I in the dosage form. In some embodiments, the average Cmax The concentration of the active ingredient in the dosage form is approximately 3 ng / mL to approximately 4 ng / mL for each mg of formula I. In some embodiments, the average C max The amount of each mg of formula I in the dosage form is in the range of 80% to 125% of 3 ng / mL to 80% to 125% of 3.5 ng / mL. In some embodiments, the dosage form has an average C of formula I for about 2 hours to about 7 hours. max The average t max It has. In some embodiments, the dosage form has the average C of formula I for about 3 hours to about 7 hours. max The average t max It has. In some embodiments, the dosage form has the average C of formula I for about 3.5 hours to about 4.5 hours. max The average t max It has. In some embodiments, the dosage form has the average C of formula I for about 5.5 hours to about 6.5 hours. max The average t max It holds.

[0018]

[0018] Further embodiments may provide a method for treating cancer in a human subject, comprising the step of administering to the subject an oral dosage form comprising a therapeutically effective amount of the compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient, wherein the compound represented by formula I is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, and the oral dosage form has an average 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 It has the mean AUC. In a further embodiment, the mean AUC 0-24 The average AUC is approximately 27 h*ng / mL to approximately 44 h*ng / mL for each mg of formula I in the dosage form. In further embodiments, the average AUC 0-24 For each mg of formula I in the aforementioned dosage form, the concentration ranges from 80% to 125% of 30 hN ng / mL to 80% to 125% of 44 hN ng / mL.

[0019]

[0019] Further embodiments provide a method for treating cancer in a human subject, comprising the step of administering to the subject an oral dosage form comprising a therapeutically effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically 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-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, and the oral dosage form is administered over an average period of about 8 to about 22 hours. 1 / 2 It has the following characteristics: In some embodiments, the average t 1 / 2 This is approximately 8 to 12 hours. In some embodiments, the average t 1 / 2 This is approximately 9 to 11 hours.

[0020]

[0020] Further embodiments provide a method for treating cancer in a human subject, comprising the step of administering to a human subject an oral dosage form comprising a therapeutically effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically 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-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, and the oral dosage form has an average 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 It has the mean AUC. In some embodiments, the mean AUC 0-inf The average AUC is approximately 29 h*ng / mL to approximately 67 h*ng / mL for each mg of formula I in the dosage form. In some embodiments, the average AUC 0-inf For each mg of formula I in the aforementioned dosage form, the concentration ranges from 80% to 125% of 36 h*ng / mL to 80% to 125% of 57 h*ng / mL.

[0021]

[0021] Further embodiments provide a method for treating cancer in a human subject, comprising the step of administering to the subject an oral dosage form comprising a therapeutically effective amount of a compound represented by formula I or a pharmaceutically acceptable salt thereof and at least one pharmaceutically 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-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, and the oral dosage form has an average 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 It has. In a further embodiment, the mean AUC 0-t For each mg of formula I in the aforementioned dosage form, the concentration ranges from 80% to 125% of 27 h*ng / mL to 80% to 125% of 36 h*ng / mL.

[0022]

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

[0023]

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

[0024]

[0024] In some embodiments reported herein, the oral dosage form is a tablet comprising an inner phase comprising a compound of formula I or a pharmaceutically acceptable salt, lactose monohydrate, low-substituted hydroxypropyl cellulose, 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 a monoHCl salt form of formula I.

[0025]

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

[0026]

[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 fasted person. In some embodiments, the oral dosage form is administered to a fed person.

[0027]

[0027] Embodiments include compound H3B-6545:

[0028]

[0028] [ka] (H3B-6545)

[0029]

[0029] A method for treating a patient with H3B-6545 or a pharmaceutically acceptable salt thereof is provided, comprising the steps of: identifying whether a decrease in one or more variant allele frequencies is observed in the patient's blood; selecting the patient if such decrease is observed; and administering a therapeutically effective amount of H3B-6545 or a pharmaceutically acceptable salt thereof to the selected patient. Variant allele frequency is the relative frequency of variant alleles (gene variants) at a particular location in a population, expressed as a fraction or percentage ratio.

[0030]

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

[0031]

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

[0032]

[0032] In some embodiments, a decrease in the frequency of variant alleles 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]

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

[0034]

[0034] In further embodiments, the reduction in mutant allele frequency is observed in ESR1. In further embodiments, the ESR1 mutation site 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 further embodiments, 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 further embodiments, the ESR1 mutation that results in the observed reduction in mutant allele frequency is S463P. In further embodiments, the ESR1 mutation that results in the observed reduction in mutant allele frequency is Y537N.

[0035]

[0035] In a further embodiment, the reduction in mutant allele frequency is observed at PIK3CA. In a further embodiment, the PIK3CA mutation site resulting 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 resulting 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]

[0036] Further embodiments include compound H3B-6545

[0037]

[0037] [ka] (H3B-6545)

[0038]

[0038] A method may be provided for discontinuing treatment with compound H3B-6545 or a pharmaceutically acceptable salt thereof from a patient receiving treatment with the compound H3B-6545 or a pharmaceutically acceptable salt thereof, the method comprising the steps of determining whether an increase in the frequency of one or more variant alleles is observed in the patient's blood, and if such increase is observed, discontinuing treatment for the patient.

[0039]

[0039] In some embodiments, an increase in the frequency of variant alleles 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]

[0040] In some embodiments, an increase in the frequency of mutant alleles is observed in ESR1. In further embodiments, the ESR1 mutation site resulting in the observed increase in the frequency of mutant alleles 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 resulting in the observed increase in the frequency of mutant alleles 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 the mutant allele frequency is observed at PIK3CA. In a further embodiment, the PIK3CA mutation site that results in the observed increase in the mutant allele frequency is selected from the group consisting of E542, E545, and H1047. In an even further embodiment, the PIK3CA mutation that results in the observed increase in the 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]

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

[0043]

[0043] In a further embodiment, the step of identifying the patient is that the compound H3B-6545: [ka] (H3B-6545)

[0044]

[0044] The treatment is performed after at least one treatment with a pharmaceutically acceptable salt thereof.

[0045]

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

[0046]

[0046] In further embodiments, an increase in the frequency of mutant alleles is observed in circulating tumor DNA. In further embodiments, a decrease in the frequency of mutant alleles is observed in circulating tumor DNA.

[0047]

[0047] The documents reported herein are incorporated by reference. In the event of any conflict between these documents and this document, this document shall prevail. [Brief explanation of the drawing]

[0048] [Figure 1] The time-course profile of the mean plasma concentration of H3B-6545 in Example 3 (Cycle 1, Day 1) is shown. Preliminary PK analysis indicates that H3B-6545 shows an increase roughly proportional to the plasma exposure dose from 100 mg to 600 mg. Tmax and half-life were considered dose and time independent. [Figure 2] The tumor response and duration of H3B-6545 treatment reported in Example 3 are shown. Partial response, stable response, progressive disease, and unassessable are indicated for each tumor evaluation. Arrows indicate treatment progression at the time of data cut. [Figure 3] This shows the percentage change in the total diameter of the target lesions reported in Example 3. [Figure 4] This diagram shows an overview of the sample sets for liquid biopsy and tissue biopsy. [Figure 5] The baseline genomics profiles of patients in Example 7 are shown. Figures 5A to 5E show the beaming assay. Figure 5A: Baseline ESR1 and PIK3CA mutation status in patients; Figures 5B and 5C: Clonality of ESR1 and PIK3CA mutations; Figures 5D and 5E: Amino acid distribution of PIK3CA and ESR1 mutations. [Figure 5F] This shows the oncoprint of mutations found in baseline tissue biopsies, as determined by the Oncomine comprehensive panel. [Figure 6A-6B] The agreement between the mutations observed in tissue biopsy and fluid biopsy in Example 7 is shown. Figure 6A shows the allele frequency distribution and agreement between the mutations in tissue biopsy and fluid biopsy. Figure 6B is a summary table of the agreement between the mutations in tissue biopsy and fluid biopsy. [Figure 7A-7C]Figure 7A shows ctDNA dynamics correlated with the overall best response to H3B-6545. AF = allele frequency, C = cycle, D = day, PD = progressive disease, PR = partial response, SD = stable disease. The dotted line represents the ddPCR assay LOD. Figure 7A shows the AF baseline / C2D1 ratio for each individually color-coded patient. A single patient may have multiple mutations. Figures 7B and 7C show exemplary ctDNA dynamics in patients with progressive disease (B) vs. partial response (C). Detailed explanation

[0049]

[0055] definition

[0050]

[0056] In this specification, the articles “a,” “an,” and “the” are considered to cover both singular and plural unless otherwise specified herein or unless otherwise clearly contradicted by the 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 forms.

[0051]

[0057] The terms “comprising,” “having,” “including,” and “containing” are open terms (i.e., “including, but not limited to”) unless otherwise stated. Furthermore, in any case where “comprising” or any other open term is used in one embodiment, it should be understood that such embodiment may be claimed in a narrower scope using the intermediate term “essentially consisting of” or the closed term “consisting of.”

[0052]

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

[0053]

[0059] As used herein, the term “mean” means the geometric mean determined from a set of independent measurements. For example, independent measurements may be collected from a statistically significant population. A further example is when used to describe pharmacokinetic parameters (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 the 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 a mean pharmacokinetic value obtained from a set of independently measured values.

[0054]

[0060] The following is a list of abbreviations and definitions of terms used in this application: AUC: Area under the plasma concentration-time curve. 0-x Area under the plasma concentration-time curve (AUC) from zero hours after administration to x hours (for example, x may represent 12 hours or 24 hours). 0-t : Area under the plasma concentration-time curve, AUC, from zero time to the last quantifiable concentration. 0-inf : Area under the plasma concentration-time curve from zero time 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 dispersion, LC-MS / MS: liquid chromatography-mass spectrometry / mass spectrometry, MAD: repeated dose escalation study, MTD: maximum tolerated dose, PD: pharmacodynamics, PK: pharmacokinetics, RT: reaction time, SAD: single dose escalation study, SD: standard deviation, t 1 / 2 : Terminal phase disappearance half-life, t max : Time to reach maximum (peak) concentration after drug administration. When used herein, t 1 / 2 This includes the terminal phase elimination half-life of the drug concentration, which is C max This may be the terminal phase disappearance half-life. When used herein, C max This includes the maximum drug concentration of the substance measured in human plasma.

[0055]

[0061] Two dosage forms with absorption rates and degrees differing by -20% / +25% or less are generally considered "bioequivalent." Another approach to mean bioequivalence involves calculating a 90% confidence interval for the ratio of the mean (population geometric mean) measurements for the test product and the reference product. To establish BE, the calculated confidence interval should typically be within the range of 80-125% for the ratio of the product mean. In addition to this general approach, other approaches, such as (1) logarithmic transformation of pharmacokinetic data, (2) methods for evaluating sequencing effects, and (3) methods for evaluating outliers, may be useful for establishing bioequivalence. For example, in (1) above, the confidence interval should typically be within the range of 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 “approximately” refer to measurable values, such as quantities and time periods, and include variations of ±20% or 10% from a given value, more preferably 5%, even more preferably 1%, and still more preferably 0.1%, because such variations are 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,” those skilled in the art will understand that providing an equivalent molar amount of the active compound as a pharmaceutically acceptable salt typically requires administering a larger mass of the salt than administering the compound alone (i.e., the amount of free base to the amount of salt has a 1:1 molar ratio). For example, the phrase “25 mg to 50 mg of the compound represented by formula I or a pharmaceutically acceptable salt thereof” refers to 25 mg to 50 mg (including the endpoint) of the free base of the compound of formula I, and 26 mg to 53 mg (including 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 coefficient conversion is also applicable to the crystalline form of formula I, which exists as a hydrate, solvate, or crystalline form, having both i) a hydrate or solvate and ii) a salt counterion. Furthermore, such potency adjustment coefficient conversion is also applicable to whether the cocrystalline solvent molecules and / or salt counterions exist in crystalline form in integral or non-integral hypothesis. Thus, those skilled in the art will understand that different potency adjustments can be made for monohydrochloride of H3B-6545, hemihydrochloride of H3B-6545, or unusual ratios, such as 1:1.3, 1:1.25, etc.

[0059]

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

[0060]

[0066] "H3B-6545 drug substance" refers to the monohydrochloride salt of H3B-6545, as reported in U.S. Patent 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 of treatment,” all of which refer to a human subject with breast cancer or a human subject at high risk of developing breast cancer relative to the general population. A human subject in need of treatment may be a human subject who has been previously diagnosed or identified as having breast cancer or a precancerous condition. Alternatively, a human subject in need of treatment may be a human subject at high risk of developing such a disorder relative to the general population (i.e., a subject who is more likely to develop such a disorder relative to the general population). A human subject in need of treatment may have a precancerous condition.

[0062]

[0068] Human subjects requiring this treatment may have refractory or resistant cancer (i.e., cancer that does not respond to treatment or has never responded to treatment). Subjects may be resistant at the start of treatment or may become resistant during treatment. In some embodiments, subjects requiring this treatment have experienced a recurrence of cancer after remission at the time of most recent treatment. In some embodiments, subjects requiring this treatment have undergone and failed all known effective treatments for cancer. In some embodiments, subjects requiring this treatment have received at least one prior treatment. In preferred embodiments, subjects have cancer or a cancerous condition.

[0063]

[0069] As used herein, “fasted state” refers to a human subject requiring fasting who has experienced at least 10 hours of fasting overnight prior to administration. Food should not be permitted for at least 4 hours after administration. Water may be permitted except for 1 hour before and after drug administration.

[0064]

[0070] As used herein, “feeding condition” refers to a human subject who has fasted overnight for at least 10 hours, consumed a high-fat breakfast within 30 minutes, and received a single dose of H3B-6545 within 30 minutes of the start of the meal. Water was permitted as desired, except for one hour before and after drug administration.

[0065]

[0071] As used herein, “treating” or “treat” means the management and care of a human subject aimed at combating a disease, condition or disorder, and includes the administration of H3B-6545 or a pharmaceutically acceptable salt, polymorph, hydrate or solvate thereof in dosage form to alleviate the symptoms or complications of such disease, condition or disorder, or to eliminate such disease, condition or disorder.

[0066]

[0072] H3B-6545 or its pharmaceutically acceptable salts and / or solvates may be used, or may be used to prevent related diseases, conditions or disorders, or to identify suitable candidates for such purposes. As used herein, “preventing,” “prevent,” or “protect against” means to reduce or eliminate the onset of symptoms or complications of such disease, condition or disorder.

[0067]

[0073] As used herein, “sample” means any biological sample derived from a human subject, including but not limited to cells, tissue samples, body 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 supplied by the subject under therapeutic or experimental conditions. Alternatively, the sample may be obtained by a physician through the usual practice of medicine in the art.

[0068]

[0074] As used herein, the term “dosage form” refers to a physically separated unit appropriate as a unit dose for a human subject being treated, each unit containing a predetermined amount of the active compound calculated to produce the desired therapeutic effect in conjunction with the necessary pharmaceutical carrier. Dosage forms are classified in terms of the route of administration and site of application, including, for example, oral, topical, rectal, vaginal, intravenous, subcutaneous, intramuscular, ocular, nasal, visual, and inhalation administration. Alternatively, dosage forms are classified in terms of the physical form, such as solid, semi-solid, or liquid. Dosage forms are any of the various forms, including, for example, IV bags, tablets, single pumps on aerosol inhalers, or vials. “Oral dosage form” refers to a dosage form that is readily administered to a human subject by mouth. Non-limiting examples of oral dosage forms include capsules and tablets. The amount of the 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 depending on the specific treatment in question.

[0069]

[0075] As used herein, the phrase “pharmaceutically acceptable” means a compound, material, composition, carrier and / or dosage form that has a suitable benefit-to-risk ratio and is appropriate for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response, or other problems or complications, within the bounds of sound medical judgment.

[0070]

[0076] As used herein, “pharmaceutically acceptable excipients” means excipients that are generally safe, non-toxic, and useful in the preparation of pharmaceutical compositions that are not biologically or otherwise undesirable, and include excipients acceptable for veterinary use and for human medicinal use. As used herein and in the claims, “pharmaceutically acceptable excipients” includes both one and more such excipients. For example, pharmaceutically acceptable excipients used in the formulation of the present invention may be diluents or inert carriers, lubricants, binders, or combinations thereof. pharmaceutically acceptable excipients used in the formulation of the present invention may further include fillers, antimicrobial agents, antioxidants, anti-caking agents, coating agents, or mixtures thereof.

[0071]

[0077] As used herein, the term “composition” includes products comprising a specific amount of a specific component and any product directly or indirectly resulting from a specific combination of the specific components. Terms relating to pharmaceutical compositions are intended to include products comprising an active ingredient (here, any of Formula I or its pharmaceutically acceptable salts, hydrates, and / or solvates) and an inert component constituting a carrier, and any product directly or indirectly resulting from any combination of two or more components, complex formation or aggregation, or dissociation of one or more components, or other types of reactions or interactions. Accordingly, the pharmaceutical compositions of the present invention include any compositions prepared by mixing a compound represented by Formula I (or its pharmaceutically acceptable salts, hydrates, and / or solvates) with a pharmaceutically acceptable excipient.

[0072]

[0078] As used herein, the term “therapeutic effective dose” refers to the amount of H3B-6545 that can produce a therapeutic effect in a human subject. A therapeutic effective dose is the amount that can treat, induce remission or prevent a specific disease or condition, or that exhibits a detectable therapeutic or inhibitory effect. The effect may be detected by any assay method known in the art. The exact effective dose for a subject depends on the subject's weight, size and health, the nature and severity of the condition, and the treatment selected for administration. A therapeutic effective dose for a given situation may be determined by routine experimental methods within the scope of the clinician’s skill and judgment.

[0073]

[0079] In a preferred embodiment, the disease or condition being treated is cancer. In another embodiment, the disease or condition being treated is a cytoproliferative disorder. A therapeutically effective dose of H3B-6545 may be administered in dosage form. A therapeutically effective dose of H3B-6545 may be in the form of a pharmaceutically acceptable salt, solvate, and / or hydrate.

[0074]

[0080] As used herein, “therapeutic effect” is the result of any type of medical treatment, and the result is judged to be desirable and beneficial. This applies whether the result is expected, unexpected, or not even intended by the treatment. Desirable or beneficial results may include inhibition of changes in cellular signaling pathways, inhibition of cell proliferation, preferably cancer cell proliferation, promotion of cell death, preferably cancer cell death, or reduction of tumor, all of which are observed without severe adverse effects. Delaying tumor growth, and preferably regression, and also preferably complete regression of the cancer, is another example of a therapeutic effect. A therapeutic effect may also be an objectively identifiable improvement recognized by a clinician or other qualified observer. For example, tumor regression in a patient may be measured by reference to the diameter of the tumor. A decrease in the diameter of the tumor indicates regression. Regression is also indicated by the non-recurrence of the tumor after discontinuation of treatment.

[0075]

[0081] Severe adverse effects include life-threatening effects (e.g., liver failure, cardiac arrhythmias, and certain types of allergic reactions), effects resulting in persistent or serious physical disability or hospitalization, or effects causing birth defects.

[0076]

[0082] As used herein, “pharmaceutically acceptable salt” refers to a derivative of H3B-6545 in which the parent compound has been modified by the preparation of its acid or base salt. Examples of pharmaceutically acceptable salts include, but are not limited to, basic residues, such as mineral or organic acid salts of amines, and acidic residues, such as alkalis or organic salts of carboxylic acids. Examples of pharmaceutically acceptable salts include 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 that it can be measured, calculated, or expressed as a quantity or numerical value. Quantifiable plasma concentration is the concentration of H3B-6545 that can be detected and measured in the plasma of a human subject after administration. Quantifiable AUC bioavailability is the fraction of H3B-6545 accessible into 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. 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 shown herein.

[0078]

[0084] As used herein, the term “solvate” means a solvent-added form containing either stoichiometric or nonstoichiometric 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 convertible from one isomeric form to the other.

[0080]

[0086] Unless otherwise stated or made clear from the context, the term “or” as used herein is understood to be inclusive.

[0081]

[0087] One aspect of the present invention provides a dosage form comprising a therapeutically effective amount of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient, which can be administered to a human subject in need thereof, wherein the therapeutically effective amount achieves a plasma concentration that can be quantified 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 yet another embodiment of the present invention, the dosage form is a solid oral dosage form. In yet another embodiment, the solid oral dosage form may be an immediate-release oral solid dosage form. The oral solid dosage form may be in the form of a tablet or a capsule. These forms may have a plurality of phases, for example, an inner phase and an outer phase.

[0082]

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

[0083]

[0089] At least one pharmaceutically acceptable excipient may be a diluent or inert carrier, a disintegrant, a lubricant, a binder, or a combination thereof. Other pharmaceutically acceptable excipients include fillers, antimicrobial agents, antioxidants, anti-caking agents, coating agents, or mixtures thereof.

[0084]

[0090] Examples of binders include corn starch, potato starch, other starches, gelatin, natural and synthetic gums, e.g., acacia, xanthan gum, sodium alginate, alginic acid, other alginates, tragacanth powder, guar gum, cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone (e.g., povidone, crospovidone, copovidone, etc.), methylcellulose, methocell, pregelatinized starch (e.g., Starch 1500® and Starch 1500 LM®, sold by Colorcon, Ltd.), hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose (FMC Corporation, Marcus Examples include, but are not limited to, 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, dextrose, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, dextrose, fructose, honey, anhydrous lactose, 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] Examples of disintegrants include, but are not limited to, agar-agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polaritrin potassium, sodium starch glycolate (e.g., Explotab), potato or tapioca starch, other starches, pregelatinized starch, clay, other algins, other celluloses, gums (e.g., gellan gum), low-substituted hydroxypropyl cellulose, polyplasdone, or mixtures thereof.

[0086]

[0092] Examples of lubricants include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, polyethylene glycol, other glycols, complitol, stearic acid, sodium lauryl sulfate, sodium stearyl fumarate (e.g., Pruv), vegetable fatty acid lubricants, talc, hydrogenated vegetable oils (e.g., peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, thyroid silica gel (AEROSIL 200, WRGrace Co., Baltimore, MD USA), synthetic silica solidification aerosol (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 phthalate acetate, ethylcellulose, gelatin, pharmaceutical glossing agents, hydroxypropylcellulose, hydroxypropyl methylcellulose (hypromellose), hydroxypropyl methylcellulose phthalate, methylcellulose, polyethylene glycol, polyvinyl acetate phthalate, shellac, sucrose, titanium dioxide, camauba wax, microcrystalline wax, gellan gum, maltodextrin, methacrylate, 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 by a coating system (e.g., the Opadry® fx film coating system) that is coated with, for example, 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 contained in a capsule having an internal phase comprising a therapeutically effective amount of H3B-6545 or a pharmaceutically acceptable salt thereof, lactose monohydrate, low-substituted hydroxypropyl cellulose, microcrystalline cellulose, hydroxypropyl cellulose, and colloidal anhydrous silica. The capsule also comprises an external phase comprising magnesium stearate.

[0090]

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

[0091]

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

[0092]

[0099] Pharmacopoecitable salts may include conventional non-toxic salts of the parent compound, or quaternary ammonium salts, formed from, for example, non-toxic inorganic or organic acids. For example, such conventional non-toxic salts include 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, Examples of salts obtained from inorganic and organic acids selected from lactobionic acid, lauryl sulfonic 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 amine acids, such as glycine, alanine, phenylalanine, and arginine, are listed, but are not limited to these.

[0093]

[0100] Other examples of pharmaceutically acceptable salts include hexanoic acid, cyclopentanepropionic acid, pyruvate, 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]-octa-2-ene-1-carboxylic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, and muconic acid. The present invention also includes salts formed when an acidic proton present in the parent compound is substituted with a metal ion, such as an alkali metal ion, an alkaline earth ion, or an ammonium ion, or when it coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, tromethamine, and N-methylglucamine. In salt form, the ratio of the compound to the cation or anion of the salt may be 1:1 or any other ratio, such as 3:1, 2:1, 1:2, or 1:3.

[0094]

[0101] In other embodiments, the dosage form may also comprise one or more active compounds (e.g., H3B-6545 or a salt thereof) in combination with at least one pharmaceutically acceptable excipient or carrier.

[0095]

[0102] Examples of solvates include, when the solvent is water, the solvate formed is a hydrate, and when the solvent is an alcohol, the solvate formed is an alcoholate. Hydrates are formed by a combination of one or more molecules of water and one molecule of a substance in which water maintains its molecular state as H2O. Hemihydrates are formed by a combination of one molecule of water and more than one molecule of a substance in which water maintains its molecular state as H2O.

[0096]

[0103] An oral dosage form comprising a therapeutically effective amount of H3B-6545, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient, which can be administered to human subjects in need thereof, may achieve quantifiable plasma concentrations approximately 0.5 to 12 hours, 0.5 to 5 hours, 0.5 to 3 hours, 1 to 5 hours, or 1 to 3 hours after administration.

[0097]

[0104] The therapeutically effective amounts of H3B-6545, or pharmaceutically acceptable salts thereof, that may be present in the oral dosage form are in the ranges of approximately 50 mg to approximately 1000 mg, approximately 100 mg to approximately 800 mg, approximately 100 mg to approximately 600 mg, approximately 200 mg to approximately 600 mg, or approximately 400 mg to approximately 600 mg.

[0098]

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

[0099]

[0106] An oral dosage form comprising a therapeutically effective amount of H3B-6545, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient, which can be administered to human subjects in need thereof, can achieve maximum quantifiable plasma concentrations of approximately 170 ng / mL to approximately 2000 ng / mL, approximately 200 ng / mL to approximately 1500 ng / mL, approximately 500 ng / mL to approximately 2000 ng / mL, approximately 500 ng / mL to approximately 1500 ng / mL, approximately 1000 ng / mL to approximately 2000 ng / mL, or approximately 1000 ng / mL to approximately 1500 ng / mL after administration.

[0100]

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

[0101]

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

[0102]

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

[0103]

[0110] A. Oral dosage form comprising approximately 100 mg to 600 mg of H3B-6545, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0104]

[0111] In some embodiments, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In such embodiments, the dose reaches the maximum quantifiable plasma concentration C after administration to a subject requiring treatment at about 150 ng / mL to about 2100 ng / mL. max(ng / mL) is achieved. In other embodiments, the dose achieves the maximum quantifiable plasma concentration after administration to subjects requiring treatment of approximately 250 ng / mL to approximately 1700 ng / mL. In further embodiments, the dose achieves the maximum quantifiable plasma concentration after administration to subjects requiring treatment of approximately 1200 ng / mL to approximately 1500 ng / mL. In these embodiments, C max This can be achieved in approximately 1 to 25 hours, 2 to 12 hours, 2 to 5 hours, or 4 hours.

[0105]

[0112] In one embodiment, the oral dosage form comprises about 100 to 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such embodiment, the oral dosage form comprises about 1.8 ng / mL to about 4.0 ng / mL of C per 1 mg of H3B-6545 max A range can be achieved. In a further embodiment, the oral dosage form contains approximately 1.8 ng / mL to approximately 3.4 ng / mL of C per 1 mg of H3B-6545. max The range can be achieved. In further embodiments, the oral dosage form contains approximately 3.0 ng / mL to approximately 4.0 ng / mL of C per 1 mg of H3B-6545. max The 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 pharmaceutically acceptable salt thereof, and at least one pharmaceutically 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 about 10.0 hours to about 13.0 hours, after administration to a human subject requiring it.

[0107]

[0114] In one embodiment, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In such embodiments, the dose can achieve quantifiable AUC bioavailability (0 to 24 hours) 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.

[0108]

[0115] In one embodiment, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such embodiments, the dose may achieve a quantifiable AUC bioavailability (0 to 24 hours) 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 the basis of "per 1 mg of H3B-6545", after administration to a human subject in need.

[0109]

[0116] In one embodiment, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In such embodiments, the dose can achieve quantifiable AUC bioavailability (0 to infinite 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.

[0110]

[0117] In one embodiment, the oral dosage form comprises about 100 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such embodiments, 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 the basis of "per 1 mg of H3B-6545" after administration to a human subject in need.

[0111]

[0118] B. Oral dosage form comprising approximately 300 mg to 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0112]

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

[0113]

[0120] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In such embodiment, the oral dosage form comprises about 3.0 ng / mL to about 4.0 ng / mL of C per 1 mg of H3B-6545. max A range can be achieved. In a further embodiment, the oral dosage form contains approximately 3.0 ng / mL to approximately 3.4 ng / mL of C per 1 mg of H3B-6545. max The 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 pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In some of these embodiments, the dose achieves a half-life of about 9.5 hours to about 14.5 hours, or about 10.0 hours to about 11.0 hours, after administration to a human subject in need.

[0115]

[0122] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In such embodiment, the dose can 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, after administration to a human subject in need.

[0116]

[0123] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In such embodiments, the dose may achieve a quantifiable AUC bioavailability (0 to 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 the basis of "per 1 mg of H3B-6545," after administration to a human subject in need.

[0117]

[0124] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient. In such embodiments, the dose can achieve quantifiable AUC bioavailability (0 to infinite 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 after administration to a human subject in need.

[0118]

[0125] In one embodiment, the oral dosage form comprises about 300 mg to about 600 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such embodiments, 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 the basis of "per 1 mg of H3B-6545" after administration to a human subject in need.

[0119]

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

[0120]

[0127] In some embodiments, the oral dosage form comprises about 450 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such embodiments, the dose is such that, after administration to a subject in need of treatment, the maximum quantifiable plasma concentration C is about 1,000 ng / mL to about 1,600 ng / mL. max(ng / mL) is achieved. 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. In these embodiments, C max This can be achieved in approximately 3 to 6 hours, or approximately 4 to 6 hours.

[0121]

[0128] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such embodiment, the oral dosage form contains about 2.4 ng / mL to about 3.5 ng / mL of C per 1 mg of H3B-6545. max A range can be achieved. In a further embodiment, the oral dosage form contains approximately 3.0 ng / mL to approximately 3.5 ng / mL of C per 1 mg of H3B-6545. max The range can be achieved. In further embodiments, the oral dosage form contains approximately 3.3 ng / mL to approximately 3.5 ng / mL of C per 1 mg of H3B-6545. max The range can be achieved.

[0122]

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

[0123]

[0130] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such embodiment, the dose may achieve 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.

[0124]

[0131] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such 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 the basis of "per 1 mg of H3B-6545", after administration to a human subject in need.

[0125]

[0132] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such embodiment, the dosage form can achieve a quantifiable AUC bioavailability (0 to infinite 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.

[0126]

[0133] In one embodiment, the oral dosage form comprises about 450 mg of H3B-6545 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient. In such 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 the basis of "per 1 mg of H3B-6545" after administration to a human subject in need.

[0127]

[0134] Embodiments may also relate to a method for treating breast cancer (i.e., ERα-positive and / or HER2-negative), the method comprising the step of administering any oral dosage form described herein to a human subject in need of it in order to obtain the described PK result.

[0128]

[0135] In all embodiments of the treatment method, the human subject requiring it may be in a state of feeding or fasting.

[0129]

[0136] Preclinical studies 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) are expected to determine the 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 depletion 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 ER covalent antagonist that inhibits both estrogen receptor α (ERα) mutant (MUT) and wild-type (WT) activity by irreversibly engaging a unique cysteine ​​not conserved in other nuclear hormone receptors. H3B-6545 potently suppresses ERα function without degrading the receptor.

[0132]

[0139] Embodiments provide a method for treating a patient with H3B-6545 or a pharmaceutically acceptable salt thereof, comprising the steps of: identifying whether a decrease in one or more variant allele frequencies is observed in the patient's blood; selecting the patient if such decrease is observed; and administering a therapeutically effective amount of H3B-6545 or a pharmaceutically acceptable salt thereof to the selected patient. Variant allele frequency is the relative frequency of variant alleles (gene variants) at a particular location in a population, expressed as a fraction or percentage.

[0133]

[0140] The inventors report a Phase 1 dose escalation 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 the H3B-6545 drug substance. The capsules used in subsequent examples were prepared using this method. Capsules with strengths of 25 mg, 50 mg, and 150 mg were formulated according to the table below.

[0136] [Table 1]

[0137] [Table 2]

[0138] [Table 3]

[0139]

[0144] In short, the capsules were prepared by forming an initial mixture of H3B-6545 drug substance, lactose monohydrate, low-substituted hydroxypropyl cellulose, microcrystalline cellulose, hydroxypropyl cellulose, and colloidal anhydrous silica in a high-shear mixer. Next, magnesium stearate was mixed with the initial mixture using the high-shear mixer. Then, the resulting mixture was compressed into a ribbon 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 Forming

[0141]

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

[0142] [Table 4]

[0143] [Table 5]

[0144]

[0147] The tablets were formed using the following method:

[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 supplied to a wet high-shear granulator, and granulation was carried out using purified water supplied by a pump to form wet granules. The wet granules were continuously dried at high temperature using a fluid bed dryer until the drying loss 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 core tablets. The coating suspension (prepared by suspending the coating agent in purified water) was then sprayed onto the core tablets using a pan coating machine.

[0148]

[0151] Blood samples were collected from each patient at the planned time and centrifuged. The plasma portion of each sample was then transferred to a K2EDTA tube and 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 analytical software.

[0149]

[0152] Blood samples (approximately 5 mL each) were collected from human patients at various time points before and after administration. Plasma concentrations of H3B-6545 were determined using verified high-performance liquid chromatography / tandem mass spectrometry (LC-MS / MS). 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 capsule or tablet form and had sufficient evaluable plasma concentration data to obtain at least one major PK parameter for each treatment. Plasma concentrations of H3B-6545 (as free base) were tabletized and summarized at each nominal time point, using descriptive statistics (number of subjects, arithmetic mean with standard deviation [SD], coefficient of variation [CV%], geometric mean, median, minimum, and maximum) as appropriate. Individual and mean (±SD) plasma concentration-time profiles were obtained for each treatment.

[0151]

[0154] The pharmacokinetic parameters of H3B-6545 were calculated using a non-compartmental method with appropriate models for plasma data and extravascular administration. PK parameters were defined as the area under the curve (AUC) up to the last quantifiable concentration. 0-t ), maximum concentration (C max ), and the point at which the maximum concentration occurs (t max This includes, but is not limited to, the data. If data is permitted, the area under the plasma concentration-time curve is estimated to infinity (AUC). 0-inf ), terminal phase disappearance half-life (t 1 / 2), apparent systemic clearance (CL / F), total systemic clearance after oral administration at steady state (CL ss / F), apparent volume of distribution during terminal phases (Vz / F), apparent steady-state volume of distribution (Vss), C max (RC max The aggregation ratio and AUC of ) 0-24 The aggregation ratio of (RAUC) was also obtained.

[0152]

[0155] Example 3: First interim analysis of capsule pharmacokinetics (PK) in fasted patients

[0153]

[0156] Example 3 reports the clinical and genomics analysis of a single trial. The study population for Example 3 was as follows: Women aged 18 or older with locally advanced or metastatic ER+HER2-BC Progression during at least one hormone therapy and at least one additional treatment / regimen, no maximum prior treatment history. ECOG performance status of 0 or 1 Sufficient bone marrow and organ function Remove if the patient has a bone-only disease or inflammatory BC.

[0154]

[0157] The method was used to analyze whether H3B-6545 inactivates both wild-type and mutant ERα by targeting cysteine ​​530 and adopting a specific antagonist conformation. Method: Women with locally progressive or metastatic HR+BC were treated with the H3B-6545 drug product administered orally once daily in capsule form over 28-day cycles, following progression at least once with hormonal therapy and at least one further treatment / regime (tx). Dose escalation was possible between patients using a 3+3 design, with the option of reinvesting previously eliminated doses. This example investigated the safety, pharmacokinetics, and pharmacodynamics of H3B-6545 in women with HR+BC to identify recommended subsequent doses for the study.

[0155]

[0158] Results: As of December 10, 2018, 32 patients were being treated with H3B-6545 drug products at doses of 100-450 mg / day. 97% had received prior treatment with CDK4 / 6 inhibitors, and 56% had received three or more lines of prior anticancer therapy. No dose-dependent toxicity was observed, and only one grade 3 treatment-related adverse event (TRAE) was observed (lymphopenia).

[0156]

[0159] The most common (over 10%) TRAEs included asymptomatic sinus bradycardia, diarrhea, nausea, fatigue, anemia, loss of appetite, and hot flashes. H3B-6545 was present for 2-4 hours. max It was rapidly absorbed. Plasma concentrations increased at doses of 100–450 mg and were comparable in C1D1 and C1D15. Consistent with the mechanism of action and preclinical data of H3B-6545, H3B-6545 inhibits ER target gene expression, showing a 50% reduction in Ki67 at all dose levels post-treatment. ESR1 (60%) and PIK3CA (34%) mutations were detected in plasma at baseline, and changes in mutant allele frequencies correlated with treatment response. Stable disease was observed in 15 patients (47%), and 34% of these patients completed at least 6 months of treatment. Partial responses (PRs) were observed in three patients: one patient (mutant) had received two lines of treatment prior to treatment; two patients (one mutant and one wild-type) had received more than five lines of treatment prior to treatment, including fulvestrant and capecitabine; and all three patients had received CDK4 / 6 inhibitors prior to treatment.

[0157]

[0160] The H3B-6545 drug product was well tolerated up to a 450 mg dose level, where initial signs of monotherapy antitumor activity were observed in past CDK4 / 6 situations. Dose escalation was continued in patients with advanced HER2-negative breast cancer.

[0158]

[0161] Drug administration and dosage: H3B-6545 was administered orally once daily over a 28-day cycle. Dose escalation was performed using a standard 3+3 study with dose cohorts of 100, 200, 300, 450, and 600 mg QD. The patient was able to reinstate the dose that had been previously eliminated. Inter-patient dose escalation was possible after the 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. 42 patients (91.3%) had previously received CDK4 / 6 inhibitors, and 32 patients (69.6%) had previously received fulvestrant. Thirty patients (65.2%) had previously received three or more lines of chemotherapy in an advanced / metastatic stage.

[0161]

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

[0162] [Table 6] TIFF0007917591000011.tif58149

[0163]

[0165] Safety: Two dose-dependent toxicities (DLTs), grade 3 fatigue and grade 3 pyolomatous measles, 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 QT prolongation on the electrocardiogram at 450 mg, anemia, increased aspartate aminotransferase, fatigue, and rash / measles at 600 mg. There were no TRAEs of grade 4 or higher. The most common TRAE, sinus bradycardia, was not considered dose- or concentration-dependent and did not require dose reduction, interruption, or discontinuation.

[0164]

[0166] TRAE is summarized in Table 7.

[0165] [Table 7]

[0166]

[0167] Pharmacokinetics: t max It took 2 to 4 hours. Plasma concentrations increased with increasing 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 the frequency of variant alleles show a correlation with clinical response.

[0168]

[0169] Patient case studies:

[0169]

[0170] A 50-year-old woman, first diagnosed with ER+HER2 breast cancer in 2006, had previously received treatments including letrozole / palbociclib, entinostat / exemestane, capecitabine, eribulin, and carboplatin / gemcitabine in a metastatic context. ESR1 Y537S and PI3KCA E545K mutations were detected in baseline tumor and plasma. At enrollment, her 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 total reduction in diameter of all target lesions from baseline to C3D1 and C7D1 was approximately -27.8% and -35.6%, respectively. After 4 cycles, the patient achieved a partial response, which was maintained with treatment at C11.

[0171]

[0172] Preliminary activity: Stable disease was observed in 17 patients (37.0%). Treatment for at least six months was completed by 14 patients (30.4%). Confirmed partial responses were observed in 3 patients (6.5%).

[0172]

[0173] Further information on preliminary activity is shown in Figure 2. Figure 3 shows the percentage change in the total diameter of the target lesions.

[0173]

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

[0174] [Table 8] TIFF0007917591000014.tif62149

[0175]

[0175] Conclusion Safety / Tolerability: Most TRAEs were Grade 1 or 2, manageable, and showed tolerance up to 450 mg QD, including 450 mg. Pharmacodynamics: Changes in mutant allele frequencies correlate with clinical responses. Preliminary activity: Signals suggesting antitumor activity were observed in heavily prior-treatment-treated patients after CDK4 / 6i, including three confirmed partial responses and mPFS = 7.2 months. Phase 2: Must begin after the RP2D decision is made.

[0176]

[0176] Example 4: Second Interim Analysis

[0177]

[0177] The clinical trial of Example 3 was continued, and the PK evaluation (of the present Example 4) was performed later based on a larger patient population. The summary in Table 1 and Table 2 of the analysis of the pharmacokinetics (PK) of the capsule in fasted patients show the PK parameters and profiles of human patients administered capsules of the H3B-6545 drug product prepared as in Example 1. Patients were administered 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 a human subject in need thereof, wherein the human subject was in a fasted state. QD means that the dose was administered "once daily (quaque die)" or only once per day. Parameter n is the number of human subjects in need thereof that were administered each dose.

[0179]

[0179] Tables 9 and 11, associated with each dose, contain the geometric mean and geometric mean coefficient of variation for all plasma PK parameters except T max and T lag , and said PK parameters are calculated from human subjects (n) in need thereof. T max , and T lag values include the minimum and maximum values, as well as the median 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 normalized to the plasma PK parameters per 1 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 Capsule in Fasting and Fed Patients

[0185]

[0181] Table 3 details the PK parameters and profiles of human patients administered the capsule prepared as in Example 1. Patients received a dose of a total equivalent of 450 mg of Formula I, as specified in Table 13. Table 14 shows the AUC exemplified in Table 13 0-t , AUC 0-24 , AUC 0-inf , and C max PK parameters are normalized to the plasma PK parameters per 1 mg of active pharmaceutical ingredient (API).

[0186] [Table 13]

[0187] [Table 14]

[0188]

[0182] Example 6: Pharmacokinetics (PK) of Tablets in Humans

[0189]

[0183] The capsule formulation described in Example 1 and the tablet formulation described in Example 2 were tested in healthy human subjects. All doses showed good tolerance. Similar exposure to H3B-6545 (C max (Both AUC and T) max This is described between capsules and tablets, as shown in Table 15.

[0190] [Table 15]

[0191]

[0184] Table 16 shows the relative bioavailability of capsule vs. tablet formulations of H3B-654.

[0192] [Table 16]

[0193]

[0185] Example 7 - Genomics Test

[0194]

[0186] Because there is no effective treatment for endocrine-resistant metastatic breast cancer (MBC), the inventors have developed H3B-6545, a novel selective Era covalent antagonist that can irreversibly inactivate both wild-type Era and mutant Era.

[0195]

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

[0196]

[0188] Method

[0197]

[0189] Fresh plasma samples were collected at baseline (pre-administration), cycle 1, day 15 (C1D15), C2D1, C3D1, and every 8 weeks thereafter, with the final sample collection at the time of disease progression. At baseline, hotspot mutations in ESR1, PIK3CA, and AKT1 were evaluated using beaming digital PCR. Patient-specific ctDNA mutations were then monitored by ddPCR. Baseline tumor biopsies were subjected to targeted next-generation sequencing (NGS) panels to identify hotspot mutations.

[0198]

[0190] Results

[0199]

[0191] In 77% (30 / 39) of patients, mutations were detected at baseline by beaming assay, with 21 / 39, 16 / 39, and 3 / 39 of these having 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 plasma beaming assay were also detected by tumor biopsy, where DNA mutations found in tissue were also detected. In 86% (12 / 14) of cases, they were detected in plasma. Continuous ctDNA monitoring revealed that in patients with confirmed partial response (3 / 3), ctDNA levels were undetectable by C2D1. In contrast, ctDNA levels were elevated from baseline in 3 / 4 of patients with progressive disease. The study shows the correlation between ctDNA ratio testing (day 15 / baseline and day 30 / baseline) and the response of PIK3CA and ESR1 mutations to H3B-6545.

[0200]

[0192] Conclusion regarding 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 metastatic situations. Furthermore, ctDNA dynamics are considered to provide a useful tool for monitoring the efficacy of H3B-6545.

[0202]

[0194] Drug administration and dosage: H3B-6545 was administered once daily (PO QD) in 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]

[0195] Liquid biopsy assay: 10 ml of Strek plasma was collected at the site and then processed in Sysmex Inostics. cfDNA was isolated from 2 ml of plasma for beaming and from 3 ml for Biorad ddPCR.

[0204]

[0196] Tumor Next-Generation Sequencing (NGS) Analysis: DNA from 10x5μm slides was isolated using the Recoverall Kit, and 10ng of DNA was used as input for the Oncomine Comprehensive Assay.

[0205]

[0197] The baseline characteristics of the study are shown in Table 17.

[0206] [Table 17]

[0207]

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

[0208]

[0199] Figures 5A to 5E show the baseline genomics profiles of patients in this embodiment. AA = amino acid. *AKT1 mutations observed in 3 out of 49 patients (data not shown).

[0209]

[0200] Figures 5A-5E show the beaming assay. A. Patient baseline ESR1 and PIK3CA mutation status, 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 agreement of mutations observed in tissue and fluid biopsies in Example 2. Figure 6A shows the allele frequency distribution and agreement of tissue and fluid biopsy mutations. Figure 6B is a table summarizing the agreement of tissue and fluid biopsy mutations.

[0212]

[0203] Figures 7A-7C show ctDNA dynamics correlated with the 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, separately colored. A single patient may have multiple mutations. Figures 7B and 7C. Exemplary ctDNA dynamics in patients with progressive disease (B) versus partial response (C).

[0213]

[0204] Patient Case Study - This is the same patient case study as reported in Example 3. A 50-year-old woman who was first diagnosed with ER+HER2-BC in 2006. Prior treatment in metastatic cases included letrozole / palbociclib, entinostat / exemestane, capecitabine, eribulin, and carboplatin / gemcitabine. At the time of registration, the patient had an ECOG performance status of 1, and disease sites included liver, bone, pleural effusion, pelvic ascites, and subcutaneous nodules. ESR1 Y537S and PI3KCA E545K mutations were detected in baseline tumor and plasma. H3B-6545 was initiated at a dose of 450 mg PO QD in June 2018. The reduction in the sum of diameters of all target lesions from baseline to C3D1 and C7D1 was -27.8% and -35.6%, respectively. After 4 cycles, the patient achieved a partial response, which was maintained during treatment at C11.

[0214]

[0205] Conclusion In late-line ER+ MBC, PIK3CA and ESR1 mutations were detected in 39% and 55% of baseline plasma samples, respectively. The concordance of tissue mutations to plasma mutations was 80%. The concordance of liquid biopsy mutations to tissue mutations was 51%. ctDNA levels were undetectable by C2D1 in patients who had confirmed partial response. ctDNA dynamics are considered to be a useful tool for monitoring the efficacy of H3B-6545 and for baseline genomic profiling.

[0215]

[0206] All publications and patent documents cited herein are incorporated by reference herein as if each such publication or document was specifically and individually indicated to be incorporated herein by reference. Citation of any publication or patent document is not intended as an admission that it is pertinent prior art, nor does it constitute an admission as to the content or date of such publication or document. The present invention has been described herein by this specification, and those skilled in the art will recognize that the present invention can be embodied in various embodiments, and that the foregoing description and examples are for illustrative purposes and are not intended to limit the scope of the following claims.

[0216]

[0207] The present invention may be embodied in other specific forms without deviating from its spirit or essential features. The embodiments described herein are therefore illustrative rather than limiting in all respects. The scope of the present invention is therefore indicated by the appended claims rather than by the foregoing description, and all variations that fall within the meaning and scope of equivalence of the claims are intended to be encompassed therein.

Claims

1. i) An internal phase comprising a compound represented by formula I or a pharmaceutically acceptable salt thereof, lactose monohydrate, low-substituted hydroxypropyl cellulose, microcrystalline cellulose, hydroxypropyl cellulose, colloidal anhydrous silica, and magnesium stearate, ii) External phase containing magnesium stearate An oral dosage form containing, The oral dosage form is a capsule, The formula I has the structure 【Chemistry 1】 The oral dosage form is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, represented by [the above].

2. The oral dosage form according to claim 1, wherein the oral dosage form includes a hypromellose capsule shell.

3. The oral dosage form according to claim 1 or 2, wherein the capsule comprises, by weight of the capsule, 25% to 30% of a compound represented by formula I or a pharmaceutically acceptable salt thereof, 10% to 15% of lactose monohydrate, 5% to 10% of low-substituted hydroxypropyl cellulose, 1% to 5% of microcrystalline cellulose, 0.5% to 5% of hydroxypropyl cellulose, 0.05% to 0.5% of colloidal anhydrous silica, and 0.1% to 1% of magnesium stearate.

4. The oral dosage form according to claim 3, wherein the capsule further comprises 40% to 45% hypromellose, 0.5% to 2% red iron oxide, and 0.5% to 2% titanium dioxide, per unit weight of the capsule.

5. i) An internal phase comprising a compound represented by formula I or a pharmaceutically acceptable salt thereof, lactose monohydrate, low-substituted hydroxypropyl cellulose, hypromellose, colloidal silicon dioxide, and purified water, ii) An outer phase containing microcrystalline cellulose and magnesium stearate An oral dosage form containing, The oral dosage form is a tablet. The formula I has the structure 【Chemistry 2】 The oral dosage form is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, represented by [the above].

6. The oral dosage form according to claim 5, further comprising a film coating containing hypromellose, talc, titanium dioxide, propylene glycol, iron(III) oxide, and purified water.

7. i) An internal phase comprising 150.00 to 160.00 mg of a compound represented by formula I or a pharmaceutically acceptable salt thereof, 67.00 to 68.00 mg of lactose monohydrate, 45.0 mg of low-substituted hydroxypropyl cellulose, 15.0 mg of microcrystalline cellulose, 9.0 mg of hydroxypropyl cellulose, 0.6 mg of colloidal anhydrous silica, and 1.5 mg of magnesium stearate, ii) An external phase containing 1.5 mg of magnesium stearate An oral dosage form containing, The oral dosage form is a capsule, The formula I has the structure 【Transformation 3】 The oral dosage form is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, represented by [the above].

8. The oral dosage form according to claim 7, wherein the oral dosage form includes a capsule shell.

9. The capsule shell is (i) 70 mg of hypromellose, (ii) Red iron oxide, and / or (iii) Titanium dioxide The oral dosage form according to claim 8, including the above.

10. i) An internal phase comprising 150.0 to 160.0 mg of a compound represented by formula I or a pharmaceutically acceptable salt thereof, 241.0 to 242.0 mg of lactose monohydrate, 90.0 mg of low-substituted hydroxypropyl cellulose, 18.0 mg of hypromellose, and 1.2 mg of colloidal silicon dioxide, ii) An outer phase containing 78.0 mg of microcrystalline cellulose and 12.0 mg of magnesium stearate An oral dosage form containing, The oral dosage form is a tablet. The formula I has the structure 【Chemistry 4】 The oral dosage form is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, represented by [the above].

11. The oral dosage form according to claim 10, wherein the oral dosage form comprises a film coating containing a coating agent and water.

12. The oral dosage form according to claim 11, wherein the film coating contains 30.0 mg of a coating agent, and the coating agent contains hypromellose, talc, titanium dioxide, propylene glycol, and iron(III) oxide.

13. i) An internal phase comprising 50.00 to 60.00 mg of a compound represented by formula I or a pharmaceutically acceptable salt thereof, 22.00 to 23.00 mg of lactose monohydrate, 15.0 mg of low-substituted hydroxypropyl cellulose, 5.0 mg of microcrystalline cellulose, 3.0 mg of hydroxypropyl cellulose, 0.2 mg of colloidal anhydrous silica, and 0.5 mg of magnesium stearate, ii) An external phase containing 0.5 mg of magnesium stearate An oral dosage form containing, The oral dosage form is a capsule, The formula I has the structure 【Transformation 5】 The oral dosage form is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, represented by [the above].

14. The oral dosage form according to claim 13, wherein the oral dosage form includes a capsule shell.

15. The oral dosage form according to claim 14, wherein the capsule shell contains hypromellose, red iron oxide, and titanium dioxide.

16. i) An internal phase comprising 25.00 to 30.00 mg of a compound represented by formula I or a pharmaceutically acceptable salt thereof, 11.00 to 12.00 mg of lactose monohydrate, 7.5 mg of low-substituted hydroxypropyl cellulose, 2.5 mg of microcrystalline cellulose, 1.5 mg of hydroxypropyl cellulose, 0.1 mg of colloidal anhydrous silica, and 0.25 mg of magnesium stearate, ii) An external phase containing 0.25 mg of magnesium stearate and An oral dosage form containing, The oral dosage form is a capsule, The formula I has the structure 【Transformation 6】 The oral dosage form is (E)-N,N-dimethyl-4-[2-[5-[(Z)-4,4,4-trifluoro-1-(3-fluoro-2H-indazole-5-yl)-2-phenylbuta-1-enyl]pyridine-2-yl]oxyethylamino]buta-2-enamide, represented by [the above].

17. The oral dosage form according to claim 16, wherein the oral dosage form includes a capsule shell.

18. The oral dosage form according to claim 17, wherein the capsule shell contains hypromellose, red iron oxide, and titanium dioxide.

19. The oral dosage form according to any one of claims 1 to 18, wherein the internal phase comprises one HCl salt form of formula I.

20. An oral dosage form according to any one of claims 1 to 19 for treating cancer in a human subject, wherein the cancer is ERα-positive breast cancer.

21. The oral dosage form according to claim 20, wherein the oral dosage form is part of a kit of two, three, or four doses of the same amount of oral dosage form for a single dose.

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