Crystalline forms of estrogen receptor antagonist

Abstract

The present disclosure provides crystalline solid forms of an estrogen receptor (ER) inhibitor, compositions thereof, and methods of treating ER-mediated conditions.

Description

[0001] Related Applications

[0002] This application claims the benefit of PCT application number PCT / CN2022 / 094231 filed on May 20, 2022, the entire contents of which are incorporated herein by reference. Background Art

[0003] Estrogen receptors (ER) play an important role in various cancers, including breast cancer. A variety of treatments targeting the estrogen receptor and / or its activity have been developed. Summary of the invention

[0004] Still need can completely inhibit estrogen receptor anti-estrogen drugs, described estrogen receptor includes by encoding estrogen receptor-α (ERα), estrogen receptor 1 (ESR1) wild type and mutant form (for example, those containing activating mutation) encoding those of gene.Selective estrogen receptor modulator (SERM) or degradation agent (SERD) is this kind of therapy particularly useful or promising tool.Recently, a kind of estrogen receptor antagonist, called complete estrogen receptor antagonist (CERAN), has become the promising therapy of completely inhibiting estrogen receptor.

[0005] Compared to other estrogen receptor antagonists, CERANs are considered "complete" because they can inactivate two different transcriptional activation functions (AF1 and AF2) of the estrogen receptor. Previous non-CERAN therapies fail when an activating mutation occurs in the gene encoding estrogen receptor 1, allowing AF1 and AF2 to be activated even in the absence of estrogen. The present disclosure provides salts, solid forms, and compositions of compounds for complete antagonism of estrogen receptors and uses thereof, providing options for the treatment of subjects suffering from cancer and / or where the subject has an estrogen receptor 1 (ESR1) mutation.

[0006] Compound (1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-1-(4-((1-propylazetidin-3-yl)oxy)phenyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole ("Compound 1"):

[0007]

[0008] is a complete estrogen receptor antagonist disclosed in PCT Publication No. WO 2017 / 059139 (the entire contents of which are incorporated herein by reference), designated as Compound B. There remains a need to identify salt, solid, hydrate and / or solvate forms of Compound 1 that can be used for various therapeutic applications.

[0009] During the process development of Compound 1, the applicant recognized that when preparing a solid dosage form (e.g., a tablet or capsule) containing Compound 1, the amorphous form of Compound 1 exhibits certain properties, such as flowability, bulk density, and handleability, which makes the process of generating a solid dosage form containing Compound 1 difficult.

[0010] In addition, some attempts to formulate Compound 1 into a form more suitable for manufacturing and formulation have instead resulted in a form in which Compound 1 is solvated with certain potentially toxic, organic solvents. Applicants recognize that both the free base and salt forms of Compound 1 exhibit a high tendency to solvate, which presents certain challenges when attempting to develop a non-solvated or hydrated form of Compound 1. As shown in Examples 6-9, polymorph screening experiments did not identify a non-solvated or hydrated form of the free base of Compound 1, and only identified a few non-solvated or hydrated salt forms of Compound 1.

[0011] However, the present disclosure solves the above problems and provides a form that, in some embodiments, exhibits desirable properties such as improved stability, hygroscopicity, flowability, compressibility, ease of processing, manufacturing consistency, particle size distribution, bulk density, pharmacokinetics, bioavailability, and ease of formulation. For example, the present disclosure encompasses the recognition that certain crystalline solid forms of Compound 1 as a fumarate salt ("Compound 1 fumarate") can be used in the compositions and methods described herein:

[0012]

[0013] In some embodiments, the present disclosure provides a non-solvated crystalline solid form of a fumarate salt of Compound 1. In some embodiments, the present disclosure provides a hydrated crystalline solid form of a fumarate salt of Compound 1.

[0014] In some embodiments, the present disclosure provides Compound 1 fumarate salt Form E, as described herein.

[0015] In some embodiments, the present disclosure provides a method of inhibiting an estrogen receptor or a mutation thereof in a biological sample, comprising contacting the biological sample with an estrogen receptor antagonist (eg, Compound 1 fumarate salt Form E).

[0016] In some embodiments, the present disclosure provides compositions comprising one or more forms of Compound 1 or Compound 1 fumarate provided herein. In some embodiments, the present disclosure provides pharmaceutical compositions comprising one or more forms of Compound 1 or Compound 1 fumarate provided herein and a pharmaceutically acceptable carrier.

[0017] In some embodiments, the present disclosure provides methods of treating a patient or subject suffering from a cancer associated with an estrogen receptor or an estrogen receptor mutation comprising administering an estrogen receptor antagonist (eg, Compound 1 fumarate salt Form E).

[0018] In some embodiments, the present disclosure provides methods of treating estrogen receptor (ER)-related diseases, disorders, and conditions (e.g., cancer) and / or otherwise modulating (e.g., inhibiting) estrogen receptors in the brain, comprising administering an estrogen receptor antagonist (e.g., Compound 1 fumarate salt Form E).

[0019] In some embodiments, the present disclosure provides methods for treating ER-related diseases, disorders, or conditions (e.g., ER-related cancers, including but not limited to tumors in the brain or cancers comprising tumors in the brain, such as brain metastases) by administering a specific full estrogen receptor antagonist (e.g., Compound 1 fumarate Form E) according to a regimen that achieves preferential accumulation in the tumor relative to the patient's plasma (i.e., accumulation in the tumor to a concentration higher than that in the plasma). BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 is the XRPD pattern of Compound 1 fumarate Form E.

[0021] Figure 2 : are the TGA curve (top) and DSC curve (bottom) of Compound 1 fumarate Form E.

[0022] Figure 3 is the XRPD pattern of Compound 1 fumarate Form E.

[0023] Figure 4 is the TGA curve of compound 1 fumarate form E.

[0024] Figure 5 is the DSC curve of Compound 1 fumarate form E.

[0025] Figure 6 is the DVS diagram of Compound 1 fumarate form E.

[0026] Figure 7 is a series of XRPD patterns from competitive slurry experiments of Compound 1 fumarate Form E and Compound 1 fumarate Form A anhydrate in isopropanol.

[0027] Figure 8 is a series of XRPD patterns from competitive slurry experiments of Compound 1 fumarate Form E and Compound 1 fumarate Form A anhydrate in water.

[0028] Fig. 9is a series of XRPD patterns from competitive slurry experiments of Compound 1 fumarate Form E and Compound 1 fumarate Form A anhydrate in ethyl acetate. DETAILED DESCRIPTION

[0029] Compound 1

[0030] Compound 1 is a full estrogen receptor antagonist disclosed in PCT Publication No. WO 2017 / 059139 (the entire contents of which are incorporated herein by reference), designated as Compound B. Exemplary methods using Compound 1 are described in PCT Publication Nos. WO 2021 / 007146 and WO 2021 / 178846, the entire contents of each of which are incorporated herein by reference.

[0031] The synthesis of compound 1 is described in detail in Example 10 of WO 2017 / 059139 and in Example 1 herein.

[0032] Compound 1 Fumarate

[0033] In some embodiments, the present disclosure provides a crystalline solid form of a fumarate salt of Compound 1, such as Compound 1 fumarate salt Form E. Compound 1 fumarate salt Form E is one of a plurality of polymorphic solid forms of a fumarate salt of Compound 1. As used herein, the term "polymorph" refers to the ability of a compound to exist in one or more different crystal structures. For example, pharmaceutically relevant physical properties of one or more polymorphs may vary between one form and another (e.g., solubility, stability, and / or hygroscopicity).

[0034] In some embodiments, Compound 1 fumarate salt Form E exists in a non-solvated form. A crystalline solid form that does not incorporate any water or solvent into the crystal structure is "non-solvated". In some embodiments, Compound 1 fumarate salt Form E exists in an anhydrous form. A crystalline solid form that does not incorporate any water into the crystal structure is an "anhydrous". In some embodiments, Compound 1 fumarate salt Form E is a non-solvated anhydrous.

[0035] In some embodiments, the crystalline form of the fumarate of Compound 1 exists in the form of a solvate and / or a hydrate. As used herein, the term "solvate" refers to a solid form in which a stoichiometric or non-stoichiometric amount of one or more solvents are incorporated into a crystal structure. For example, a solvated or heterogeneously solvated polymorph may independently contain 0.05, 0.1, 0.2, 0.5, 1.0, 1.5, 2.0 equivalents of one or more solvents incorporated into the crystal lattice. As used herein, the term "hydrate" refers to a solvate in which the solvent incorporated into the crystal structure is water.

[0036] It is understood that "Compound 1 fumarate" refers to a complex form comprising Compound 1 non-covalently associated with the co-former fumaric acid. Such non-covalent associations include, for example, ionic interactions, dipole-dipole interactions, π-stacking interactions, hydrogen bonding interactions, etc. It is understood that the term "Compound 1 fumarate" encompasses salt forms resulting from ionic interactions between Compound 1 and fumaric acid as well as non-ionic associations between Compound 1 and fumaric acid.

[0037] As provided herein, Compound 1 fumarate salt Form E has distinct XRPD peaks not previously reported in disclosures of Compound 1. As used herein, the term "about" when used in reference to a degree 2-theta value refers to the state value ± 0.2 degrees 2-theta.

[0038] In some embodiments, the present disclosure provides a complex form comprising Compound 1 and fumaric acid (i.e., Compound 1 fumarate), wherein the complex form is Compound 1 fumarate Form E. In some embodiments, Compound 1 fumarate Form E comprises a 1:1 ratio of fumaric acid to Compound 1. In some embodiments, Compound 1 fumarate Form E is an anhydrate.

[0039] In some embodiments, the provided forms (e.g., forms of Compound 1 and Compound 1 fumarate) are characterized by having peaks in their XRPD patterns selected from "substantially all" of the provided lists, optionally within ±0.2 degrees 2θ of the specified values. It should be understood that an XRPD pattern of "substantially all" of the provided lists of peaks refers to an XRPD pattern containing at least 80% (e.g., 80%, 85%, 90%, 95%, 99%, or 100%) of the listed peaks. In some embodiments, the XRPD pattern contains at least 90% of the listed peaks. In some embodiments, the XRPD pattern contains all of the listed peaks. In some embodiments, the XRPD pattern contains all of the listed peaks except one. In some embodiments, the XRPD pattern contains all of the listed peaks except two. In some embodiments, the XRPD pattern contains all of the listed peaks except three.

[0040] In some embodiments, the provided forms (e.g., forms of Compound 1 and Compound 1 fumarate) are characterized by having a "substantially similar" figure or spectrum to the drawings provided herein. It should be understood that a figure or spectrum having "substantially similar" to the drawings provided herein is a figure or spectrum containing one or more features of the provided drawings (e.g., position (degree 2θ) values, temperature values, weight loss % values, intensity, curve shape, etc.) so that it can be identified that the form characterized by the figure or spectrum (e.g., solid and / or salt form) is the same as the form characterized in the drawings. For example, in some embodiments, an XRPD figure having substantial similarity to the provided drawings is a figure containing substantially all of the same peaks, optionally within a range of ±0.2 degrees 2θ of the peaks in the reference drawings. In some embodiments, an XRPD figure having substantial similarity to the provided drawings is a figure containing substantially all of the same peaks, optionally within a range of ±0.2 degrees 2θ of the peaks in the reference drawings, with approximately the same intensity.

[0041] In some embodiments, Compound 1 fumarate salt Form E is characterized by one or more peaks in its XRPD pattern selected from about 5.83, about 7.03, about 8.69, about 12.88, about 13.43, about 14.68, about 15.65, about 16.65, and about 18.46 degrees 2θ. In some embodiments, Compound 1 fumarate salt Form E is characterized by two or more peaks in its XRPD pattern selected from about 5.83, about 7.03, about 8.69, about 12.88, about 13.43, about 14.68, about 15.65, about 16.65, and about 18.46 degrees 2θ. In some embodiments, Compound 1 fumarate salt Form E is characterized by three or more peaks in its XRPD pattern selected from those at about 5.83, about 7.03, about 8.69, about 12.88, about 13.43, about 14.68, about 15.65, about 16.65, and about 18.46 degrees 2θ.

[0042] In some embodiments, Compound 1 fumarate salt Form E is characterized by peaks at about 5.83, about 7.03, about 8.69, about 12.88, about 13.43, about 14.68, about 15.65, about 16.65, and about 18.46 degrees 2θ in its XRPD pattern. In some embodiments, Compound 1 fumarate salt Form E is characterized by peaks in its XRPD pattern at substantially all of the following positions:

[0043]

[0044]

[0045] In some embodiments, Compound 1 fumarate salt Form E is characterized by one or more of the following:

[0046] (i) with Figure 1 and / or Figure 3 An XRPD pattern substantially similar to the XRPD pattern depicted in;

[0047] (ii) with Figure 2 and / or Figure 4 A TGA graph substantially similar to the TGA graph depicted in; and

[0048] (iii) Figure 2 and / or Figure 5 The DSC graph depicted in is essentially similar to the DSC graph depicted in .

[0049] Prepare the provided form

[0050] In some embodiments, the present disclosure provides a method for preparing a provided solid form (e.g., Compound 1 fumarate Form E). In some embodiments, Compound 1 fumarate Form E is prepared by contacting Compound 1 (e.g., amorphous Compound 1, crystalline Compound 1, or a mixture thereof) with fumaric acid. In some embodiments, the present disclosure provides a method for preparing Compound 1 fumarate Form E, comprising the steps of: providing Compound 1; and combining Compound 1 with fumaric acid (optionally in a suitable solvent) to provide Compound 1 fumarate Form E. In some embodiments, about 1.0, about 1.1, about 1.2, or about 2.0 equivalents of fumaric acid are added.

[0051] In some embodiments, Compound 1 fumarate Form E is prepared by dissolving Compound 1 fumarate (e.g., amorphous Compound 1 fumarate, crystalline Compound 1 fumarate, or a mixture thereof) in a suitable solvent and then restoring Compound 1 fumarate to a solid phase. In some embodiments, Compound 1 fumarate Form E is prepared by mixing Compound 1 fumarate (e.g., amorphous Compound 1 fumarate, crystalline Compound 1 fumarate, or a mixture thereof) in a suitable solvent under suitable conditions and isolating Compound 1 fumarate Form E.

[0052] In some embodiments, the suitable solvent is selected from 2-butanol, ethylene dichloride, ethanol, heptane, isopropanol, N-methylpyrrolidone, and water, or any combination thereof.

[0053] In some embodiments, the method for preparing Compound 1 fumarate Form E comprises the step of heating a mixture comprising a fumarate salt of Compound 1 to a suitable temperature (e.g., about 30°C to about 60°C). In some embodiments, the method for preparing Compound 1 fumarate Form E comprises the step of stirring a mixture comprising a fumarate salt of Compound 1 at ambient temperature. In some embodiments, the method for preparing Compound 1 fumarate Form E comprises the step of cooling a mixture comprising a fumarate salt of Compound 1 to a suitable temperature (e.g., about -20°C to about 0°C).

[0054] In some embodiments, Compound 1 fumarate Form E is precipitated from a mixture (e.g., a solution, suspension, or slurry). In some embodiments, Compound 1 fumarate Form E is crystallized from a solution. In some embodiments, Compound 1 fumarate Form E is crystallized from a solution after solution seeding (e.g., adding Compound 1 fumarate Form E to a solution). In some embodiments, Compound 1 fumarate Form E is precipitated or crystallized from a mixture after cooling, adding an antisolvent, and / or removing all or part of the solvent by a method such as evaporation, distillation, filtration, reverse osmosis, absorption, or reaction.

[0055] In some embodiments, the method for preparing Compound 1 Fumarate Form E includes the step of isolating Compound 1 Fumarate Form E. It should be understood that Compound 1 Fumarate Form E can be isolated by any suitable means. In some embodiments, Compound 1 Fumarate Form E is separated from the supernatant by filtration. In some embodiments, Compound 1 Fumarate Form E is separated from the supernatant by decantation.

[0056] In some embodiments, the isolated Compound 1 fumarate salt Form E is dried (eg, in air or under reduced pressure, optionally at elevated temperature).

[0057] In some embodiments, Compound 1 fumarate Form E is prepared by converting a solid form of Compound 1 fumarate to Compound 1 fumarate Form E.

[0058] In some embodiments, Compound 1 fumarate form E is prepared by a process comprising the following steps: Compound 1 (e.g., amorphous Compound 1) is combined in a suitable solvent (e.g., isopropanol) at a suitable temperature (e.g., about 40°C) under stirring. In some embodiments, the process further comprises adding a first portion (e.g., about 0.5 equivalents) of fumaric acid. In some embodiments, the process further comprises adding seeds of Compound 1 fumarate form E. In some embodiments, the process further comprises adding a second, third, and / or fourth portion (e.g., about 0.2-0.3 equivalents) of fumaric acid. In some embodiments, the process further comprises adding a suitable antisolvent (e.g., heptane). In some embodiments, the process further comprises cooling the mixture to ambient temperature (e.g., about 25°C). In some embodiments, the process further comprises separating the solid form of Compound 1 fumarate form E by a method such as filtration.

[0059] Composition

[0060] In some embodiments, the present disclosure also provides compositions comprising Compound 1 fumarate salt Form E.

[0061] In some embodiments, the composition provided comprising Compound 1 fumarate Form E is substantially free of impurities. As used herein, the term "substantially free of impurities" means that the composition does not contain a large amount of foreign matter. Such foreign materials may include starting materials, residual solvents, or any other impurities that may be produced by the preparation and / or separation of a crystalline solid form. In some embodiments, the composition comprises at least about 90% by weight of Compound 1 fumarate Form E. In some embodiments, the composition comprises at least about 95% by weight of Compound 1 fumarate Form E. In some embodiments, the composition comprises at least about 99% by weight of Compound 1 fumarate Form E.

[0062] In some embodiments, the compositions provided comprising compound 1 fumarate form E are substantially pure (e.g., comprising at least about 95%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5% or 99.8% by weight of the crystalline solid form based on the total weight of the composition). In some embodiments, the composition comprising compound 1 fumarate form E comprises no more than about 5.0% of total organic impurities. In some embodiments, the composition comprising compound 1 fumarate form E comprises no more than about 3.0% of total organic impurities. In some embodiments, the composition comprising compound 1 fumarate form E comprises no more than about 1.5% of total organic impurities. In some embodiments, the composition comprising compound 1 fumarate form E comprises no more than about 1.0% of total organic impurities. In some embodiments, the composition comprising compound 1 fumarate form E comprises no more than about 0.5% of total organic impurities. In some embodiments, the percentage of total organic impurities is measured by HPLC.

[0063] In some embodiments, the composition comprises compound 1 fumarate form E and an amorphous solid form (e.g., an amorphous solid form of compound 1 and / or a fumarate of compound 1). In some embodiments, the composition comprising a crystalline solid form is substantially free of an amorphous solid form. As used herein, the term "substantially free of an amorphous solid form" means that the composition does not contain a significant amount of an amorphous solid form. In some embodiments, the composition comprises at least about 90% by weight of compound 1 fumarate form E. In some embodiments, the composition comprises at least about 95% by weight of compound 1 fumarate form E. In some embodiments, the composition comprises at least about 99% by weight of compound 1 fumarate form E. In some embodiments, the composition comprises no more than about 10% by weight of an amorphous solid form (e.g., an amorphous solid form of compound 1 and / or a fumarate of compound 1). In some embodiments, the composition comprises no more than about 5% by weight of an amorphous solid form (e.g., a crystalline solid form of compound 1 and / or a fumarate of compound 1). In some embodiments, the composition comprises no more than about 1% by weight of an amorphous solid form (eg, a crystalline solid form of Compound 1 and / or Compound 1 fumarate salt).

[0064] Pharmaceutical composition

[0065] In some embodiments, the present disclosure provides a pharmaceutical composition comprising Compound 1 fumarate salt Form E and a pharmaceutically acceptable carrier.

[0066] In some embodiments, provided pharmaceutical compositions contain an amount of Compound 1 (e.g., in the form of Compound 1 fumarate Form E) that is effective for measurably inhibiting estrogen receptor (ER) or a mutant thereof in a biological sample or patient. In some embodiments, provided pharmaceutical compositions are formulated for oral administration.

[0067] In some embodiments, provided pharmaceutical compositions comprise Compound 1 fumarate salt Form E and one or more fillers, disintegrants, lubricants, glidants, anti-adherents and / or antistatic agents, and the like.

[0068] The pharmaceutical compositions of the present disclosure can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, intraperitoneally, intracisternal, or via an implanted reservoir. In some embodiments, the pharmaceutical compositions provided are administered orally, intraperitoneally, or intravenously. In some embodiments, the pharmaceutical compositions provided are administered orally.

[0069] In some embodiments, the pharmaceutical composition provided is an oral dosage form (e.g., a capsule or tablet). In some embodiments, the pharmaceutical composition provided is a tablet. In some embodiments, the pharmaceutical composition provided is a capsule.

[0070] In some embodiments, provided pharmaceutical compositions are solid pharmaceutical compositions (eg, solid dosage forms such as capsules or tablets).

[0071] In some embodiments, the pharmaceutical compositions provided include an amount of Compound 1 suitable for providing to humans that is equivalent to a dose of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 mg / kg of Compound 1 in mice. In some embodiments, the pharmaceutical compositions provided include an amount of Compound 1 suitable for providing to humans that is equivalent to a dose of at least 3 mg / kg of Compound 1 in mice. In some embodiments, the pharmaceutical compositions provided include an amount of Compound 1 suitable for providing to humans that is equivalent to a dose of at least 5 mg / kg of Compound 1 in mice. In some embodiments, the pharmaceutical compositions provided include an amount of Compound 1 suitable for providing to humans that is equivalent to a dose of at least 10 mg / kg of Compound 1 in mice. In some embodiments, the pharmaceutical compositions provided include an amount of Compound 1 suitable for providing to humans that is equivalent to a dose of at least 15 mg / kg of Compound 1 in mice. In some embodiments, provided pharmaceutical compositions comprise an amount of Compound 1 suitable for providing to humans that is equivalent to a dose of at least 20 mg / kg of Compound 1 in mice. In some embodiments, provided pharmaceutical compositions comprise an amount of Compound 1 suitable for providing to humans that is equivalent to a dose of at least 25 mg / kg of Compound 1 in mice. In some embodiments, provided pharmaceutical compositions comprise an amount of Compound 1 suitable for providing to humans that is equivalent to a dose of at least 30 mg / kg of Compound 1 in mice.

[0072] In some embodiments, the provided pharmaceutical composition is administered once a day (QD). In some embodiments, the provided pharmaceutical composition is administered twice a day (QD). In some embodiments, the provided pharmaceutical composition is administered every other day (QOD). In some embodiments, the provided pharmaceutical composition is administered once a week (QW). In some embodiments, the provided pharmaceutical composition is administered once every four weeks (Q4W).

[0073] In some embodiments, provided pharmaceutical compositions (e.g., unit dosage forms) contain about 15 mg to about 120 mg of Compound 1. In some embodiments, provided pharmaceutical compositions (e.g., unit dosage forms) contain about 15 mg to about 100 mg of Compound 1. In some embodiments, provided pharmaceutical compositions (e.g., unit dosage forms) contain about 60 mg to about 120 mg of Compound 1. In some embodiments, provided pharmaceutical compositions (e.g., unit dosage forms) contain about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, or about 100 mg of Compound 1. In some embodiments, provided pharmaceutical compositions (e.g., unit dosage forms) contain about 15 mg of Compound 1. In some embodiments, provided pharmaceutical compositions (e.g., unit dosage forms) contain about 30 mg of Compound 1. In some embodiments, provided pharmaceutical compositions (e.g., unit dosage forms) contain about 60 mg of Compound 1. In some embodiments, provided pharmaceutical compositions (e.g., unit dosage forms) contain about 90 mg of Compound 1. In some embodiments, provided pharmaceutical compositions (eg, unit dosage forms) comprise about 120 mg of Compound 1. In some embodiments, the unit dosage form is a capsule. In some embodiments, the unit dosage form is a tablet.

[0074] It should be understood that reference to the amount of Compound 1 (e.g., in mg) in connection with, for example, a pharmaceutical composition, a dosing regimen, etc., refers to the amount of Compound 1 in free base form. Thus, Compound 1 may be provided and / or utilized, for example, in salt form, such that the amount of the salt (or other form) is an amount corresponding to a "free base equivalent" of Compound 1.

[0075] In some embodiments, provided pharmaceutical compositions are prepared by: (i) providing Compound 1 fumarate salt Form E; and (ii) formulating Compound 1 fumarate salt Form E with a suitable excipient to provide a pharmaceutical composition.

[0076] use

[0077] The compounds and compositions described herein are generally useful for inhibition of estrogen receptors (ER) and their mutants. In some embodiments, the present disclosure encompasses the insight that the compounds and compositions described herein are useful for treating ER-related disorders (e.g., ER-related cancers, such as breast cancer, including metastatic brain cancer), detecting ER-related diseases, and / or identifying certain tumors.

[0078] For example, in some embodiments, the present disclosure provides certain treatment methods for subjects suffering from ER-related diseases, disorders, or conditions. In some embodiments, the ER-related diseases, disorders, or conditions are cancer. In some embodiments, the ER-related diseases, disorders, or conditions are selected from breast cancer, bone cancer, lung cancer, colorectal cancer, endometrial cancer, prostate cancer, ovarian cancer, vaginal cancer, endometriosis, and uterine cancer. In some embodiments, the ER-related diseases, disorders, or conditions are breast cancer.

[0079] In some embodiments, the subject has been determined to have or is suspected of having a cancer that has metastasized (e.g., to the brain, bones, lungs, liver, or central nervous system). In some embodiments, the subject has been determined to have or is suspected of having brain metastases. In some embodiments, the subject has developed brain metastases associated with ER-related cancers (e.g., breast cancer) or estrogen receptor mutations.

[0080] In some embodiments, the methods provided include administering Compound 1 (e.g., as Compound 1 fumarate Form E) to a subject previously treated with an ER inhibitor. In some such embodiments, the methods provided include administering Compound 1 (e.g., as Compound 1 fumarate Form E) to a subject previously treated with a selective estrogen receptor modulator (SERM), including, for example, tamoxifen, endoxifene, raloxifene, toremifene, lasofoxifene, and ospemifene.

[0081] In some embodiments, the methods provided include administering Compound 1, or a crystalline form or complex form thereof, to a subject suffering from an ER-related disorder (e.g., breast cancer) who is unresponsive to SERM therapy, including, for example, tamoxifen, endoxifen, raloxifene, toremifene, lasofoxifene, and ospemifene.

[0082] In some embodiments, the subject relapses during or after treatment with a SERM, including, for example, tamoxifen, endoxifen, raloxifene, toremifene, lasofoxifene, and ospemifene.

[0083] In some embodiments, the methods provided include administering Compound 1 (e.g., as Compound 1 fumarate Form E) to a subject with an estrogen receptor-positive (ER+) and human epidermal growth factor receptor-negative (HER-) disease. In some embodiments, the methods provided include administering Compound 1 (e.g., as Compound 1 fumarate Form E) to a subject with an estrogen receptor-positive (ER+) and human epidermal growth factor receptor-positive (HER+) disease.

[0084] In some embodiments, the amount of compound 1 (e.g., as compound 1 fumarate form E) administered to a subject is about 15 mg to about 360 mg. In some embodiments, the amount of compound 1 administered to a subject is about 30 mg to about 360 mg. In some embodiments, the amount of compound 1 administered to a subject is about 30 mg to about 300 mg. In some embodiments, the amount of compound 1 administered to a subject is about 60 mg to about 120 mg. In some embodiments, the amount of compound 1 administered to a subject is about 15 mg to about 100 mg. In some embodiments, the amount of compound 1 administered to a subject is about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, or about 100 mg. In some embodiments, the amount of compound 1 administered to a subject is about 120 mg, about 150 mg, about 210 mg, or about 300 mg. In some embodiments, the amount of compound 1 administered to a subject is about 30 mg. In some embodiments, the amount of Compound 1 administered to a subject is about 60 mg. In some embodiments, the amount of Compound 1 administered to a subject is about 90 mg. In some embodiments, the amount of Compound 1 administered to a subject is about 120 mg.

[0085] In some embodiments, the amount of Compound 1 (e.g., as Compound 1 fumarate Form E) administered to a subject is about 15 mg to about 360 mg per day (QD). In some embodiments, the amount of Compound 1 administered to a subject is about 30 mg to about 360 mg per day (QD). In some embodiments, the amount of Compound 1 administered to a subject is about 30 mg to about 300 mg per day (QD). In some embodiments, the amount of Compound 1 administered to a subject is about 60 mg to about 120 mg per day (QD). In some embodiments, the amount of Compound 1 administered to a subject is about 15 mg to about 100 mg QD. In some embodiments, the amount of Compound 1 administered to a subject is about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, or about 100 mg QD. In some embodiments, the amount of Compound 1 administered to a subject is about 120 mg, about 150 mg, about 210 mg, or about 300 mg QD. In some embodiments, the amount of Compound 1 administered to a subject is about 30 mg QD. In some embodiments, the amount of Compound 1 administered to a subject is about 60 mg QD. In some embodiments, the amount of Compound 1 administered to a subject is about 90 mg QD. In some embodiments, the amount of Compound 1 administered to a subject is about 120 mg QD.

[0086] In some embodiments, compound 1 (e.g., as compound 1 fumarate form E) is administered to a subject in a unit dosage form. In some embodiments, the unit dosage form is a capsule or tablet. In some embodiments, the unit dosage form comprises about 15 mg to about 120 mg of compound 1. In some embodiments, the unit dosage form comprises about 15 mg to about 100 mg of compound 1. In some embodiments, the unit dosage form comprises about 60 mg to about 120 mg of compound 1. In some embodiments, the unit dosage form comprises about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, or about 100 mg of compound 1. In some embodiments, the unit dosage form comprises about 15 mg of compound 1. In some embodiments, the unit dosage form comprises about 30 mg of compound 1. In some embodiments, the unit dosage form comprises about 60 mg of compound 1. In some embodiments, the unit dosage form comprises about 90 mg of compound 1. In some embodiments, the unit dosage form comprises about 120 mg of compound 1. In some embodiments, the unit dosage form is a capsule. In some embodiments, the unit dosage form is a tablet.

[0087] In some embodiments, the total daily dose of compound 1 administered to a subject is about 15 mg to about 360 mg per day (QD). In some embodiments, the total daily dose of compound 1 administered to a subject is about 30 mg to about 360 mg. In some embodiments, the total daily dose of compound 1 administered to a subject is about 30 mg to about 300 mg. In some embodiments, the total daily dose of compound 1 administered to a subject is about 60 mg to about 120 mg. In some embodiments, the total daily dose of compound 1 administered to a subject is about 15 mg to about 100 mg QD. In some embodiments, the total daily dose of the amount of compound 1 administered to a subject is about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, or about 100 mg QD. In some embodiments, the total daily dose of compound 1 administered to a subject is about 120 mg, about 150 mg, about 210 mg, or about 300 mg. In some embodiments, the total daily dose of Compound 1 administered to a subject is about 30 mg QD. In some embodiments, the total daily dose of Compound 1 administered to a subject is about 60 mg. In some embodiments, the total daily dose of Compound 1 administered to a subject is about 90 mg. In some embodiments, the total daily dose of Compound 1 administered to a subject is about 120 mg.

[0088] Combination therapy

[0089] The present disclosure encompasses the recognition that certain combinations of agents can be advantageously used to fully antagonize estrogen receptors. Thus, in some embodiments, the present disclosure provides methods for treating subjects suffering from ER-related disorders (e.g., cancer, such as breast cancer), comprising administering a full estrogen receptor antagonist and a secondary anticancer agent. For example, in some embodiments, the full estrogen receptor antagonist is Compound 1 (e.g., as Compound 1 fumarate Form E). In some embodiments, the secondary anticancer agent is a CDK 4 / 6 inhibitor, a PI3KCA inhibitor, or an mTOR inhibitor.

[0090] In some embodiments, the present disclosure provides a method for treating a patient or subject suffering from cancer, the method comprising administering a complete estrogen receptor antagonist (e.g., Compound 1, e.g., as Compound 1 fumarate form E) and a CDK4 / 6 inhibitor (i.e., an agent that inhibits one or both of CDK4 and CDK6). In some embodiments, the anticancer agent is a CDK4 / 6 inhibitor selected from palbociclib, ribociclib, abemaciclib, leroxilib, treraciclib, and SHR6390. In some embodiments, the CDK4 / 6 inhibitor is palbociclib. In some embodiments, the CDK4 / 6 inhibitor is ribociclib. In some embodiments, the CDK4 / 6 inhibitor is abemaciclib. In some embodiments, the CDK4 / 6 inhibitor is leroxilib. In some embodiments, the CDK4 / 6 inhibitor is treraciclib. In some embodiments, the CDK 4 / 6 inhibitor is SHR6390.

[0091] In some embodiments, the present disclosure provides a method for treating a patient or subject suffering from cancer, the method comprising administering a complete estrogen receptor antagonist (e.g., Compound 1, e.g., as Compound 1 fumarate form E) and a PIK3CA inhibitor. In some embodiments, the PIK3CA inhibitor is selected from alpelisib, taselisib, and LY3023414. In some embodiments, the PIK3CA inhibitor is alpelisib. In some embodiments, the PIK3CA inhibitor is taselisib. In some embodiments, the PIK3CA inhibitor is LY3023414.

[0092] In some embodiments, the present disclosure provides a method for treating a patient or subject suffering from cancer, the method comprising administering a complete estrogen receptor antagonist (e.g., Compound 1, e.g., as Compound 1 fumarate form E) and an mTOR inhibitor. In some embodiments, the mTOR inhibitor is selected from sirolimus, temsirolimus, everolimus, and LY3023414. In some embodiments, the mTOR inhibitor is sirolimus. In some embodiments, the mTOR inhibitor is temsirolimus. In some embodiments, the mTOR inhibitor is everolimus. In some embodiments, the mTOR inhibitor is LY3023414.

[0093] In some embodiments, the present disclosure provides methods of treating subjects with ER+ and HER+ diseases with a full estrogen receptor antagonist (e.g., Compound 1, e.g., as Compound 1 fumarate Form E) and a HER2 inhibitor. In some embodiments, the HER2 inhibitor is selected from tucatinib, pertuzumab, lapatinib, trastuzumab, ado-trastuzumab emtansine, trastuzumab, detrastuzumab, and neratinib.

[0094] It should be understood that the combination therapy comprising a complete estrogen receptor antagonist and an anticancer agent as described herein may include administering the agent simultaneously or separately. For example, in some embodiments, a complete estrogen receptor antagonist and an anticancer agent are administered simultaneously. In some embodiments, the anticancer agent is administered before administering the complete estrogen receptor antagonist. In some embodiments, the anticancer agent is administered after administering the complete estrogen receptor antagonist.

[0095] Example

[0096] The examples provided herein record and support certain aspects of the present disclosure, but are not intended to limit the scope of any claim. The following non-limiting examples are provided to further illustrate certain teachings provided by the present disclosure. According to the application, it will be understood by those skilled in the art that various changes may be made to the specific embodiments described in the present example without departing from the spirit and scope of this teaching.

[0097] In the examples below, the following abbreviations may be used: aq. (aqueous solution); ACN (acetonitrile); CSA (camphorsulfonic acid); d (day); DCM (dichloromethane); DEA (diethylamine); DHP (dihydropyran); DMF (N,N-dimethylformamide); DIPEA (N,N-diisopropylethylamine); DMAP (4-dimethylaminopyridine); DMSO (dimethyl sulfoxide); EA (ethyl acetate); ee (enantiomeric excess); equiv. (equivalent); ethanol (EtOH); h (hour); Hex (hexane); HPLC (high performance liquid chromatography); IPA (isopropanol); KHMDS (bis(trimethylsilyl) (potassium amide); LAH (lithium aluminum hydride); LCMS (liquid chromatography-mass spectrometry); LDA (lithium diisopropylamide); LiHMDS (lithium bis(trimethylsilyl)amide); MeOH (methanol); min (minute); NMR (nuclear magnetic resonance); Pd / C (palladium on carbon); PPh3O (triphenylphosphine oxide); Pt / C (platinum on carbon); rb (round bottom); Rf (retention factor); rt or RT (room temperature); SM (starting material); TEA (triethylamine); THF (tetrahydrofuran); THP (tetrahydropyran); TLC (thin layer chromatography); TsOH (p-toluenesulfonic acid or p-toluenesulfonic acid); and UV (ultraviolet).

[0098] Materials and methods

[0099] X-ray powder diffraction (XRPD)

[0100] XRPD was performed using a PANalytical X'Pert PRO MPD or Empyrean diffractometer using an incident Cu radiation beam generated by an Optix long microfocus source. An elliptically graded multilayer mirror was used to focus the Cu Kα X-rays through the sample and to the detector. Prior to analysis, a silicon sample (NISTSRM 640f) was analyzed to verify that the position of the observed Si 111 peak was consistent with the NIST certified position. The sample was sandwiched between Kapton films and analyzed in transmission geometry. A beam cutoff, a short anti-scatter extender, and an anti-scatter knife edge were used to minimize the background generated by air. Soller slits were used for both the incident and diffracted beams to minimize the broadening caused by axial divergence. Diffraction patterns were collected using a scanning position sensitive detector (X'Celerator) located 240 mm from the sample and Data Collector software v.5.5.

[0101] Alternatively, XRPD was performed using a Bruker D8 Focus X-ray diffractometer equipped with a LynxEye detector. The sample scan range was 3° to 42° (2θ) with a step size of 0.02° (2θ). The tube voltage and current were 40 KV and 40 mA, respectively.

[0102] Thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC)

[0103] TGA / DSC analysis was performed using a Mettler-Toledo TGA / DSC3+ analyzer. Temperature and enthalpy adjustments were made using indium, tin, zinc, aluminum, gold, and phenyl salicylate, and then demonstrated with indium. Balance was demonstrated with calcium oxalate. The sample was placed in an open aluminum pan, hermetically sealed, the lid pierced, and then inserted into the TG furnace. A weighed aluminum pan (configured as a sample pan) was placed on a reference platform. The furnace was heated under nitrogen.

[0104] Alternatively, TGA was performed using a TGA Q500 (TA Instruments, US). About 1-5 mg of sample was placed in an open, tared aluminum pan, automatically weighed, and inserted into a TGA furnace. The sample was heated to a final temperature (about 300° C.) at a rate of 10° C. / min. DSC characterization was performed on a DSC 250 (TA Instruments, US). About 1-5 mg of sample was placed in a DSC pinhole pan. The sample was heated to a final temperature (about 300° C.) at a rate of 10° C. / min. The change in heat flux with temperature was recorded.

[0105] Dynamic Vapor Sorption (DVS)

[0106] DVS was performed using Intrinsic DVS (System Measurement System, UK). Approximately 30-50 mg of sample was placed in a sample basket and suspended in the measurement chamber. For isotherm testing, the chamber temperature was maintained at a constant 25±1°C by a water bath. The samples were tested in step mode with a target RH of 0 to 90% full cycle. The analysis was performed in 10% RH increments. The duration of each RH was set to 60 min to allow the sample to reach equilibrium with the chamber environment. Data were collected in 20 s increments.

[0107] Gas chromatography (GC)

[0108] GC analysis was performed on a GC8890 (Agilent, US) using helium as carrier gas, nitrogen as make-up gas, and an FID detector. The sample was 10 mg / mL in dimethylacetamide. The vaporized sample was carried to the chromatographic column by the carrier gas (mobile phase). The parameters are summarized as follows:

[0109]

[0110] Example 1: Synthesis of Compound 1

[0111] A complete synthesis of Compound 1 is provided in PCT Publication No. WO 2017 / 059139, which is incorporated herein by reference and repeated below.

[0112] Preparation of 4-((1-propylazetidin-3-yl)oxy)benzaldehyde:

[0113]

[0114] Step 1: Preparation of 1-propionylazetidin-3-one

[0115]

[0116] Compound 3-azetidinone hydrochloride (10.000g, 93.0mmol, 1.0 equivalent), anhydrous 1,2-dichloroethane (200mL) and diisopropylethylamine (38.9mL, 223mmol, 2.4 equivalents) are added to a round-bottom flask (500mL) to obtain a light yellow suspension. The suspension is ultrasonically treated for 1h and then cooled to -10°C (dry ice / MeOH) for 10min. Propionyl chloride (9.8mL, 112mmol, 1.2 equivalents) is added dropwise to the cooled suspension to obtain an orange solution. The reaction is removed from the bath and stirred at room temperature for 16h. The solvent is removed to obtain a semi-solid. The semi-solid is suspended in EA (300mL) and the suspension is filtered. The solid is rinsed with EA (2x 100mL). TLC analysis (10% MeOH / DCM, KMnO7 stain / heat) indicated three spots: Rf: 0.2, 0.5, 0.7. TLC (50% EA / Hex, KMnO7 stain / heat) indicated two spots: Rf: 1, 0.3. The filtrate was concentrated, adsorbed on silica gel (25 g), and chromatographed over silica gel (100 g small column) for 15 min with DCM (5 minutes) and then 0-10% MeOH. The product was initially isolated from the column in DCM and continued to be eluted from the column with up to 10% MeOH. TLC was performed in both solvent systems to determine if there was any propionyl chloride in the early fractions. The fractions containing the product were combined and concentrated to give the title compound as a yellow liquid (11.610 g, 98.2%).

[0117] 1 H NMR (300MHz, CDCl3) δ: 4.80 (d, J = 5.6Hz, 4H), 2.29 (q, J = 7.5Hz, 2H), 2.01 (s, 3H), 1.18 (t, J = 7.5Hz, 3H).

[0118] Step 2. Preparation of 1-propylazetidin-3-ol

[0119]

[0120] Lithium aluminum hydride (10.397 g, 273.9 mmol, 3.0 equivalents) was suspended in THF (200 mL) and cooled in an ice bath. A solution of 1-propionylazetidin-3-one (11.610 g, 91.3 mmol, 1.0 equivalent) in THF (100 mL) was added dropwise to the reaction mixture through an equalizing funnel over 30 min. The addition funnel was removed. A condenser was then installed on the flask and the reaction was heated to reflux in an oil bath at 75 ° C for 16 h. The reaction was cooled in an ice bath for 20 min, and sodium sulfate decahydrate (glauber's salt, 25 g) was added in small portions over 20 min. After the addition was complete, the mixture was stirred at room temperature for 2 h. The mixture was stirred by The mixture was filtered through a bed of 4% paraffin (2 cm) and the solid was rinsed with EA (2 x 250 mL). The clear solution was concentrated to a light yellow liquid (9.580 g, 91.1%). NMR indicated the presence of THF and EA. The material was used to prepare the compounds of the following examples without further purification.

[0121] 1 H NMR (300 MHz, CDCl3) δ: 4.39 (quintet, J = 6 Hz, 1H), 3.62-3.56 (m, 2H), 2.90-2.85 (m, 2H), 2.41 (t, J = 7.5 Hz, 2H), 1.34 (sextet, J = 7.2 Hz, 2H), 0.87 (t, J = 7.8 Hz, 3H).

[0122] Step 3. Preparation of 4-((1-propylazetidin-3-yl)oxy)benzaldehyde

[0123]

[0124] Use Teflon TMStir bar 4-fluorobenzaldehyde (15.00g, 120.9mmol, 0.9 equivalent), 1-propylazetidin-3-ol (15.00g, 130.2mmol, 1.0 equivalent), cesium carbonate (88.40g, 271.3mmol, 2.1 equivalent) and N, N-dimethylformamide (284mL) were mixed in a 500mL round-bottom flask. The flask was sealed and heated at 95°C in a heating block for 6h. The reaction was analyzed by LCMS to indicate that the aldehyde was consumed. The suspension was filtered through a sintered glass funnel and the solid was washed with ethyl acetate (100mL). The filtrate was concentrated to an orange suspension. The suspension was mixed with water (200mL) and ethyl acetate (200mL), and the organic layer was washed with water (3x 200mL), brine, dried over anhydrous magnesium sulfate, filtered and concentrated to an orange liquid (21.74g, 76.1%). This material was used without further purification.

[0125] 1 HNMR (300 MHz, CDCl3), δ9.87 (s, 1H), 7.82 (d, J = 9.0 Hz, 2H), 6.86 (d, J = 8.7 Hz, 2H), 4.86 (quintet, J = 5.7 Hz, 1H), 3.85-3.80 (m, 2H), 3.13-3.08 (m, 2H), 2.48 (t, J = 7.2 Hz, 2H), 1.46-1.34 (m, 2H), 0.91 (t, J = 7.2 Hz, 3H).

[0126] Preparation of (R)-1-(1H-indol-3-yl)-N-((R)-1-phenylethyl)propan-2-amine:

[0127]

[0128] Indole-3-acetone (25.0 g, 144 mmol, 1.0 equiv) was added to a solution of (R)-(+)-1-phenylethylamine (23.0 mL, 181 mmol, 1.3 equiv) in dichloromethane (600 mL) under N2 at 25 °C, and the mixture was stirred for 1 h. The reaction was cooled to 0-5 °C, and sodium triacetoxyborohydride (100 g, 472 mmol, 3.3 equiv) was added to the ice-cooled solution via a powder addition funnel within 30 minutes. The orange solution was stirred at 0 °C for 1 h, and then allowed to warm to RT. The reaction was stirred at RT for 19 h. At this point, ESI+ indicated the absence of indole starting material. Under vigorous stirring, saturated NaHCO3 solution (100 mL) was added in 5 mL portions at 10 °C over 15 min. The solution was stirred for 15 min and saturated Na2CO3 solution (200 mL) was added over 15 minutes. Solid K2CO3 (9g) was added in 3g portions, at which point the pH of the aqueous layer was 12 and no more bubbles were formed. The layers were filtered and separated. The red organic layer was washed with saturated NaHCO3 aqueous solution (2x 100mL). The aqueous layers were combined and extracted with DCM (2x100mL). The combined organic layers were dried over Na2SO4, filtered and concentrated to give a crude product (49g). TLC (90:10DCM:MeOH) showed four spots (Rf=0.63, 0.50, 0.16, 0.26), two of which were separated diastereoisomers (Rf=0.16 and 0.26). The crude product was adsorbed onto silica gel and purified by flash chromatography (330g small column, 0-100% EA:Hex). The fractions containing R,R diastereoisomers were combined and purified for a second time using the same flash chromatography conditions to give 24g of product (about 82% ee). The previous successful separation was achieved with a 40:1 silica gel:crude ratio, so the mixture was divided into 3 portions and separated on 3x 330g silica gel cartridges (0-40% EA / Hex 20min, isocratic 40% EA / Hex 40min). All fractions containing the desired product were >99% diastereoisomerically pure. The pure fractions were concentrated and combined to give (R)-1-(1H-indol-3-yl)-N-((R)-1-phenylethyl)-propan-2-amine as an orange semi-solid (11.91 g, 29.6%).

[0129] 1H NMR (CDCl3, 300 MHz) R, R diastereomers: δ 0.96 (d, J = 6.6 Hz, 3H), 1.30 (d, J = 6.6 Hz, 3H), 2.68 (q, J = 7.2 Hz, 1H), 2.97 (m, 2H) 4.00 (q, J = 6.3 Hz, 1H), 7.43-6.97 (m, 10H), 7.96 (br s, 1H). R, S diastereomers: δ 1.11 (d, J = 5.7 Hz, 3H), 1.30 (d, J = 5.4 Hz, 3H) 2.80 (m, 3H), 3.92 (q, J = 6.9 Hz, 1H), 6.93-7.40 (m, 10H), 8.13 (br s, 1H); due to insufficient purity, the aromatic region is difficult to distinguish from the R, R diastereomer.

[0130] LCMS: ES+ [M+H]+ 279.0.

[0131] Preparation of (2R)-1-(1H-indol-3-yl)propan-2-amine:

[0132]

[0133] Compound (R)-1-(1H-indol-3-yl)-N-((R)-1-phenylethyl)propan-2-amine (11.91 g, 42.8 mmol, 1.0 equiv) was dissolved in methanol (250 mL) and added to a 2L Parr bottle. The solution was bubbled with N2 for 10 min. 20% Pd(OH)2 on carbon wetted with water (10.71 g, 76.3 mmol, 1.8 equiv) was added, the bottle was pressurized with 50 psi hydrogen, and shaken in a Parr apparatus for 22 h. LCMS analysis indicated that the reaction was complete. The suspension was passed through The crude product was dissolved in DCM and washed with saturated Na2CO3 solution (50 mL), and the aqueous layer was extracted with DCM (2 x 50 mL). The organic layers were combined, dried and concentrated to give (2R)-1-(1H-indol-3-yl)propan-2-amine as a light brown solid without further purification (6.68 g, 89.6%).

[0134] 1H NMR (CDCl3, 300 MHz) δ 1.17 (d, J = 6.6 Hz, 3H), 2.66 (dd, J = 8.4, 14.7 Hz, 1H), 2.88 (dd, J = 5.4, 14.1 Hz, 1H), 3.27 (sextet, J = 1.5 Hz, 1H), 7.05-7.22 (m, 3H), 7.37 (d, J = 7.5 Hz, 1H), 7.62 (d, J = 8.7 Hz, 1H), 8.00 (br s, 1H).

[0135] LCMS: ES+[M+H]+174.9.

[0136] Preparation of 2-fluoro-2-methylpropanol:

[0137]

[0138] Methyl 2-fluoro-2-methylpropanoate (5.01 g, 40.5 mmol, 1.0 eq) was added dropwise over 15 min to a stirred suspension of lithium aluminum hydride (2.50 g, 65.9 mmol, 1.6 eq) in anhydrous ether (100 mL) cooled in an ice bath. After 2 h, 2.0 mL of water, 2.0 mL of 15% w / v NaOH, and 5.0 mL of water were added dropwise. After 15 min, the white suspension was diluted with DCM and washed with water by Gravity filter and wash the solid with DCM.The filtrate was concentrated (200 mbar, 25 °C) to give 2-fluoro-2-methylpropanol as a colorless oil (2.09 g, 56.1%).

[0139] 1 H NMR (300MHz, CDCl3) δ1.34 (d, J = 21.3Hz, 6H), 1.95 (br t, 1H), 3.56 (dd, J = 6.6, 20.7Hz, 2H).

[0140] Preparation of 2-fluoro-2-methylpropyl trifluoromethanesulfonate:

[0141]

[0142] Trifluoromethanesulfonic anhydride (5.0 mL, 29.7 mmol, 1.3 eq) was added dropwise to a 0°C solution of 2-fluoro-2-methylpropanol (2.090 g, 22.7 mmol, 1.0 eq) and 2,6-lutidine (3.40 mL, 29.4 mmol, 1.3 eq) in DCM (25 mL). After 2 hours, the red solution turned light brown. TLC (20:80 EA:Hex, KMnO4 staining) indicated that the starting material was absent. The reaction mixture was washed with 1 M HCl solution (2 x 20 mL) and saturated NaHCO3 solution (2 x 20 mL). The aqueous layer was back extracted with DCM (20 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure (150 mbar, 25°C) to give 2-fluoro-2-methylpropyl trifluoromethanesulfonate as a red oil (4.39 g, 86.3%).

[0143] 1 H NMR (300MHz, CDCl3) δ1.46 (d, J = 20.4Hz, 6H), 4.41 (d, J = 18.6Hz, 2H). 19 F NMR (282MHz, CDCl3) δ-147.1,-74.5.

[0144] Preparation of (R)-N-(1-(1H-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-1-amine:

[0145]

[0146] The compound 2-fluoro-2-methylpropyl trifluoromethanesulfonate (9.587 g, 42.8 mmol, 1.1 eq) (solution in DCM, 16% DCM wt%, 11.4384 g) was added to a solution of (2R)-1-(1H-indol-3-yl)propan-2-amine (6.680 g, 38.3 mmol, 1.0 eq), anhydrous 1,4-dioxane (60.000 ml, 701.4 mmol, 18.3 eq) and freshly distilled diisopropylethylamine (8.500 ml, 48.8 mmol, 1.3 eq). The dark brown solution was heated at 90 °C for 3 hours. After 3 h, LCMS indicated that a small amount of indoleamine starting material was still present. TLC (10% MeOH / DCM) indicated that the triflate (Rf=0.54) had been consumed. The NMR of the unused triflate SM (286-30) indicated that the triflate did not decompose after overnight, so another 0.1 equivalent (0.9883 g, 13% DCM weight %, 0.8563 g triflate SM) was added and the reaction was heated at 90° C. for 2 h. LCMS indicated that the reaction was complete and TLC (10% MeOH / DCM) showed one spot (Rf=0.24) (TLC with 50% EA / Hex, 1 streak-like spot Rf<=0.12, another spot at Rf=0). EtOAc (50 mL) was added, the solution was washed with NaHCO (2× 50 mL), and the combined aqueous layers were washed with EtOAc (50 mL). The combined organic extracts were dried over NaSO and concentrated under reduced pressure. The crude product (brown oil, 14.8 g) was purified by flash silica gel chromatography (240 g column, 0-100% EA / Hex). The desired product eluted as a long tailing peak. The pure fractions were concentrated to give (R)-N-(1-(1H-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-1-amine (4.211 g, 17.0 mmol) as a dark yellow oil.

[0147] 1 H NMR (300MHz, CDCl3) δ1.10 (d, J=6.3Hz, 3H), 1.34 (dd, J=3.0, 21.9Hz, 6H), 2.68-2.95 (m, 4H), 3.02 (sextet, J= 6.6Hz,1H),7.05(d,J=2.4Hz,1H),7.26-7.11(m,2H),7.36(d,J=6.9Hz,1H),7.62(d,J=7.5Hz,1H),8.18(br s,1H). 19 F NMR (282MHz, CDCl3) δ-144.2.m / z: ES+[M+H]+249.0.

[0148] Preparation of compound 1

[0149] 4-((1-propylazetidin-3-yl)oxy)benzaldehyde (0.096 g, 0.4 mmol, 1.3 eq) was added to a solution of (R)-N-(1-(1H-indol-3-yl)propan-2-yl)-2-fluoro-2-methylpropan-1-amine (0.070 g, 0.3 mmol, 1.0 eq) in anhydrous toluene (1.50 mL) and glacial acetic acid (0.100 mL, 1.7 mmol, 6.2 eq). Molecular sieves were added and the solution was stirred at 80 °C under N2 and darkness for 8 hours. The reaction solution was diluted in DCM, filtered, and washed with saturated Na2CO3 solution. The aqueous layer was extracted with DCM and the combined organic layers were dried over Na2SO4. The solution was filtered and concentrated. The residue was dissolved in acetonitrile (2 mL), filtered through a syringe filter, and then purified by preparative LC (40 to 90% ACN:H2O over 18 min, then isocratic 90% ACN for 7 min). The pure fractions were concentrated and dried to give (1R,3R)-2-(2-fluoro-2-methylpropyl)-3-methyl-1-(4-((1-propylazetidin-3-yl)oxy)phenyl)-2,3,4,9,-tetrahydro-1H-pyrido[3,4-b]indole as a white powder.

[0150] 1 H NMR(300MHz, CDCl3)δ0.90(t,J=7.5Hz,3H),1.09(d,J=7.2Hz,3H),1.26-1.50(m,8H),2.45-2.77(m,6H),3.01(t,J=7.2Hz,2H),3.34 (m,1H),3.77(m,2H),4.60(quin,J=5.7Hz,1H),5.03(s,1H),6.64(d,J=8.1Hz,2H),7.10-7.21(m,5H),7.54(d,J=7.5Hz,1H),8.19(br s,1H).m / z:ES+[M+H]+450.2.

[0151] Example 2: Preparation and Characterization of Compound 1 Fumarate Form E

[0152] Compound 1 fumarate form E is obtained according to the following exemplary procedure: Compound 1 fumarate form A (about 30-100 mg) is slurried in isopropanol at ambient temperature for 20 days. The solid is separated to obtain compound 1 fumarate form E. The preparation of compound 1 fumarate form A is as follows: fumaric acid (52.6 mg) is weighed and placed in a 20-mL glass vial. A 40 mg / mL ethyl acetate solution (15 mL) of amorphous compound 1 is added to the vial, and the mixture is stirred at RT. The sample collected after stirring for 1 day is confirmed to be compound 1 fumarate form A by XRPD. The resulting suspension is filtered, and the wet cake is vacuum dried at 50 ° C for 5h. Collect solids to obtain compound 1 fumarate form A (231.9 mg, about 92.2% yield).

[0153] The XRPD pattern of compound 1 fumarate form E is as follows Figure 1 shown.

[0154] like Figure 2 As shown in the DSC curve in , the sample shows an endothermic peak at 147°C (onset temperature) / 156°C (peak temperature). Figure 2 Also shown is a TGA curve which shows no weight loss up to 180° C. Compound 1 fumarate salt Form E was determined to be unsolvated.

[0155] 1 HNMR analysis indicated that the stoichiometry of fumaric acid:Compound 1 was approximately 1:1.

[0156] Compound 1 fumarate form E is also prepared as follows: amorphous compound 1 (80.5 mg) and fumaric acid (24.4 mg) are mixed and suspended in isopropanol (2 mL) under stirring on a magnetic stirrer. Heptane (1 mL) is added to the clear solution, and the sample is placed in a refrigerator. After about one day, the solid is separated by centrifugal filtration and analyzed by XRPD.

[0157] Compound 1 fumarate form E is also prepared as follows: amorphous compound 1 (2.0 g) and isopropanol (30 mL) are loaded into a 50 mL reactor vessel. Compound 1 is dissolved at 40 ° C, and the stirring rate is maintained at 300 rpm (two-blade paddle). Fumaric acid (0.5 equivalent) is added and dissolved after stirring for 5 min. Then the seeds of compound 1 fumarate form E (1.0 weight %) are added. After stirring for 1 h, fumaric acid (0.2 equivalent) is added. After stirring for another 1 h, fumaric acid (0.2 equivalent) is added. After stirring for another 1 h, fumaric acid (0.3 equivalent) is added. Then, heptane (30 mL) is added within 4 h. The mixture is kept at 40 ° C for 1 h, then cooled to 25 ° C within 3 h, and then stirred for 10 h. The suspension is filtered, and the wet cake is dried in a vacuum oven at 40 ° C for 16 h to obtain compound 1 fumarate form E (2.2 g, 90% yield).

[0158] Compound 1 fumarate form E is also prepared as follows: Amorphous compound 1 (20.0 g) and isopropanol (300 mL) are loaded into a 1000 mL reactor vessel. Compound 1 is dissolved at 40 ° C, and the stirring rate is maintained at 300 rpm (retreat curve impeller, RCI). Fumaric acid (0.5 equivalent) is added and stirred for 20 min. Then, a seed of compound 1 fumarate form E (1.0 wt %) is added. After stirring for 1 h, fumaric acid (0.2 equivalent) is slowly added. After stirring for another 1 h, fumaric acid (0.2 equivalent) is slowly added. After stirring for another 1 h, fumaric acid (0.3 equivalent) is slowly added. Then, heptane (300 mL) is added within 4 h. The mixture is kept at 40 ° C for 1 h, then cooled to 25 ° C within 3 h, and then stirred for 10 h. The suspension was filtered and the wet cake was dried in a vacuum oven at 40 °C for 16 h to give Compound 1 fumarate salt Form E (23.2 g, 92% yield). XRPD analysis ( Figure 3 ) confirmed that the substance was compound 1 fumarate form E. TGA analysis ( Figure 4 ) shows a weight loss of 0.438% starting from 151.2°C, DSC ( Figure 5 ) showed a melting point of 157.9°C (peak temperature). Gas chromatography (GC) indicated an isopropanol content of 3264 ppm and a heptane content of 1434 ppm. DVS analysis ( Figure 6 ) indicates that Compound 1 fumarate salt Form E is not hygroscopic.

[0159] Exemplary XRPD data for Compound 1 fumarate salt Form E are summarized below:

[0160]

[0161]

[0162] Example 3: Solubility studies

[0163] A solubility study of Compound 1 and Compound 1 fumarate in different solvents at 25° C. was performed. The solubility was measured by both kinetic and gravimetric methods.

[0164] Dynamic method: Under the conditions of a certain amount of solute and a certain temperature, gradually add solvent under stirring for 15 minutes to reach equilibrium. When the solute is completely dissolved, record the amount of solvent and calculate the solubility.

[0165] Gravimetric method: Add excess solid and a certain amount of solvent to an 8 mL vial, stir for 24 hours, and filter. Take 1 mL of the clear upper liquid, dry at 50 °C for 24 hours, weigh, and calculate the solubility.

[0166] Compound 1 fumarate form A anhydrous is prepared as follows: amorphous compound 1 (1.0 g) and ethyl acetate (25 mL) are added to a 50 mL reactor vessel. Compound 1 is dissolved at 25 ° C, and the stirring rate is maintained at 300 rpm (two-blade paddle). Fumaric acid (1.2 equivalents) is added once. After stirring for 10 min, a large amount of solids are precipitated. The mixture is stirred for 15 h. The suspension is filtered, and the wet cake is dried in a vacuum oven at 40 ° C for 16 h to obtain compound 1 fumarate form A anhydrous (1.035 g, 82% yield).

[0167] The results are summarized in Table 1. Amorphous Compound 1 has high solubility in isopropanol, and fumaric acid has relatively low solubility in isopropanol. Compound 1 fumarate form E has extremely low solubility in isopropanol at 25°C, and the solubility decreases significantly with increasing heptane ratio. The solubility of Compound 1 fumarate form E is much lower than that of Compound 1 fumarate form A anhydrate, indicating that form E is more stable than form A.

[0168] Table 1

[0169]

[0170] Example 4: Bulk density study

[0171] The bulk density of Compound 1 Fumarate Form E was measured by gently introducing a known sample mass into a graduated cylinder (50 mL), smoothing the powder without compacting it, and recording the apparent untapped volume to the nearest scale unit. The experiment was repeated three times and the results are summarized in Table 2.

[0172] Table 2

[0173] run Bulk density (g / mL) 1 0.1288 2 0.1233 3 0.1196 average value 0.1239

[0174] Example 5: Stability Study

[0175] Amorphous Compound 1, Compound 1 Fumarate Form E, and Compound 1 Fumarate Form A Anhydrate were placed in an oven at 60°C for two weeks to evaluate their stability. As shown in Table 3, Compound 1 Fumarate Form E and Compound 1 Fumarate Form A Anhydrate exhibited improved stability relative to amorphous Compound 1.

[0176] Table 3

[0177]

[0178] A competitive slurry experiment was conducted with Compound 1 Fumarate Form E and Compound 1 Fumarate Form A Anhydrate as follows:

[0179] Experiment #1: Compound 1 fumarate Form E (100 mg), Compound 1 fumarate Form A anhydrate (100 mg) and isopropanol (2 mL) were added to two 8-mL vials and mixed in an oscillator at 25°C and 40°C, respectively. Samples were collected for analysis at 24h and 72h. The XRPD analysis results are shown in Figure 7 After 24 h at 25°C or 40°C, Compound 1 fumarate Form A anhydrate was completely converted to Compound 1 fumarate Form E, indicating that Form E is more stable than Form A anhydrate in isopropanol.

[0180] Experiment #2: Compound 1 Fumarate Form E (100 mg), Compound 1 Fumarate Form A Anhydrate (100 mg) and water (2 mL) were added to two 8-mL vials and mixed in an oscillator at 25°C and 40°C, respectively. Samples were collected for analysis at 24h and 72h. The XRPD analysis results are shown in Figure 8 After 72 h at 25°C or 40°C, a mixture of Compound 1 Fumarate Form E and Compound 1 Fumarate Form A anhydrate remained, indicating that the conversion between the forms is very slow in water.

[0181] Experiment #3: Compound 1 fumarate Form E (100 mg), Compound 1 fumarate Form A anhydrate (100 mg) and ethyl acetate (2 mL) were added to two 8-mL vials and mixed in an oscillator at 25°C and 40°C, respectively. Samples were collected for analysis at 24h and 72h. The XRPD analysis results are shown in Fig. 9 After 72 h at 25°C or 40°C, a mixture of Compound 1 fumarate Form E and Compound 1 fumarate Form A anhydrate remained, indicating that the conversion between the forms was slow in ethyl acetate.

[0182] Example 6: Polymorph Screening of Compound 1

[0183] Starting from amorphous compound 1, polymorph screening of compound 1 was performed under 100 experimental conditions. A total of eight screening methods were used, including antisolvent addition, reverse antisolvent addition, 5°C slurry, RT slurry, slow evaporation, slow cooling, temperature cycling, and solid vapor diffusion. Polymorph screening identified at least two crystalline forms of compound 1, both of which were solvates. Form A was found to exist in the form of a variety of different isostructural solvates (e.g., acetonitrile, acetone, and tetrahydrofuran solvates). Form B was determined to be a DMSO solvate. The results are summarized in Table 4, as shown below:

[0184] Table 4

[0185] method Number of experiments Identified crystal forms Anti-solvent addition 8 Amorphous Slow evaporation 9 Form A and amorphous Slow cooling 11 Form A and amorphous RT sizing 30 Form A, Form B and amorphous Slurry at 5℃ 11 Form A and amorphous Solid vapor diffusion 15 Form A, Form B and amorphous Reverse antisolvent addition 8 Amorphous Temperature Cycle 8 Form A and amorphous total 100

[0186] Antisolvent addition experiments were performed under 8 conditions. About 15 mg of compound 1 was dissolved in 0.4-3.0 mL of solvent to obtain a clear solution. The solution was magnetically stirred, and then 0.1 mL of antisolvent was added in each step of the first 1 mL, and 0.5 mL was added stepwise until precipitation occurred or the total amount of antisolvent reached 5.0 mL. The resulting precipitate was isolated for XRPD analysis. As summarized in Table 5, only amorphous compound 1 was observed.

[0187] Table 5

[0188]

[0189]

[0190] Slow evaporation experiments were performed at RT under 9 different conditions. About 15 mg of compound 1 was dissolved in 0.5 mL of solvent. All solutions and suspensions were filtered using a 0.45 μm PTFE membrane and the filtrate was used in the following steps. The visually clear solution was covered with an HPLC cap, a hole was pierced in the cap with a pipette tip, and then evaporated at room temperature. The isolated solid was used for XRPD analysis. The results summarized in Table 6 show that Form A was obtained under specific conditions:

[0191] Table 6

[0192] Solvent (v / v) Observation form ACN Form A MeOH Amorphous EtOH Amorphous acetone Amorphous THF gel IPA gel EtOAc gel <![CDATA[CHCl3]]> gel DCM gel

[0193] Slow cooling experiments were performed in 11 solvent systems. About 20 mg of compound 1 was dissolved in 1.0-2.0 mL of solvent at 60 ° C and filtered into a new vial using a 0.45 μm PTFE membrane. The filtrate was slowly cooled from 60 ° C to 5 ° C at a rate of 0.05 ° C / min. The resulting solid was kept isothermal at 5 ° C and then separated for XRPD analysis. An antisolvent was added to the clear solution to cause precipitation. If no solid was observed after adding the antisolvent, slow evaporation was performed. The results summarized in Table 7 indicate that Form A was obtained under specific conditions:

[0194] Table 7

[0195] Solvent (v / v) Observed form ACN Form A <![CDATA[MeOH / H2O(1:4)]]> Amorphous* <![CDATA[EtOH / H2O(1:4)]]> Amorphous* <![CDATA[Acetone / H2O (1:4)]]> Form A+Amorphous* <![CDATA[ACN / H2O(1:4)]]> Form A <![CDATA[THF / H2O(1:4)]]> Form A+Amorphous* <![CDATA[MeOH / H2O(1:9)]]> Amorphous* <![CDATA[EtOH / H2O(1:9)]]> Amorphous* <![CDATA[Acetone / H2O (1:9)]]> Amorphous* <![CDATA[ACN / H2O(1:9)]]> Form A <![CDATA[THF / H2O(1:9)]]> Amorphous*

[0196] *Use slow evaporation procedure as described above.

[0197] Slurry conversion experiments were performed in 30 solvent systems at RT. About 20 mg of Compound 1 was suspended in 0.3 mL of solvent at RT for 4 days. The remaining solid was separated for XRPD analysis. The results summarized in Table 8 indicate that Form A and Form B were obtained under specific conditions:

[0198] Table 8

[0199]

[0200]

[0201] *Theoretical water activity based on software simulation.

[0202] Slurry conversion experiments were performed at 5°C in 11 solvent systems. Approximately 30 mg of Compound 1 was suspended in 0.3 mL of solvent at 5°C for 4 days. The remaining solid was isolated for XRPD analysis. The results summarized in Table 9 indicate that Form A was obtained under specific conditions:

[0203] Table 9

[0204]

[0205] *Use slow evaporation program.

[0206] Solid vapor diffusion experiments were performed using 12 different solvents. About 15 mg of compound 1 was weighed into a 3-mL vial, which was placed in a 20-mL vial containing 4 mL of a volatile solvent. The 20 mL vial was sealed with a cap and stored at RT for 9 days to allow the solvent vapor to interact with the sample. The solid was tested by XRPD. The results summarized in Table 10 show that Form A and Form B were obtained under specific conditions:

[0207] Table 10

[0208] Solvents Observation form <![CDATA[H2O]]> Amorphous MeOH Amorphous EtOH gel IPA gel acetone gel MEK gel MIBK gel IPA gel THF gel 2-MeTHF gel MTBE gel DCM gel ACN Form A Toluene gel DMSO Form B

[0209] In 8 solvent systems, reverse antisolvent addition experiments were performed by first placing 1 mL of antisolvent in a 3-mL glass vial in a 5°C refrigerator. Approximately 10 mg of compound 1 was then dissolved in 1 mL of solvent in a 2-mL glass vial. After the suspension was magnetically stirred for 2 hours to obtain a clear solution, the solution was quickly filtered into a 5°C antisolvent. The sample was then placed at 5°C for crystallization. If no crystallization occurred after 1 day, the sample was moved to -20°C for precipitation. The remaining solid was separated for XRPD analysis. The results summarized in Table 11 indicate that only amorphous compound 1 was obtained:

[0210] Table 11

[0211] Solvents Solvent resistance Observation form MeOH <![CDATA[H2O]]> Amorphous EtOH <![CDATA[H2O]]> Amorphous IPA <![CDATA[H2O]]> Amorphous acetone <![CDATA[H2O]]> Amorphous THF <![CDATA[H2O]]> Amorphous ACN <![CDATA[H2O]]> Amorphous DMSO <![CDATA[H2O]]> Amorphous DMF <![CDATA[H2O]]> Amorphous

[0212] Temperature cycling experiments were performed in 8 solvent systems. About 20 mg of compound 1 was suspended in 0.1 mL of solvent in a 23-mL glass vial at RT. The suspension was then heated to 60 ° C and balanced for two hours. The slurry was slowly cooled to 5 ° C at a rate of 0.1 ° C / min, and then heated to 60 ° C within one hour. The cycle was repeated once more, and then cooled to 5 ° C at a rate of 0.1 ° C / min. The sample was stored at 5 ° C, and the solid was then separated and analyzed using XRPD. The results summarized in Table 12 show that Form A was obtained.

[0213] Table 12

[0214] Solvent (v / v) Observation form ACN Form A <![CDATA[H2O]]> Amorphous <![CDATA[EtOH / H2O(1:4)]]> Amorphous <![CDATA[DMSO / H2O(1:4)]]> Amorphous <![CDATA[ACN / H2O(aw=0.2,989 / 11)]]> Form A <![CDATA[ACN / H2O(aw=0.4,978 / 22)]]> Form A <![CDATA[ACN / H2O(aw=0.6,959 / 41)]]> Form A <![CDATA[ACN / H2O(aw=0.8,925 / 75)]]> Form A

[0215] Example 7: Salt screening of compound 1

[0216] Salt screening was performed at room temperature (RT). A total of 100 salt screening experiments were performed using 25 acids in 4 different solvent systems. Specifically, the stock solutions of Compound 1 are summarized in Table 13. A summary of the salt screening is presented in Table 14.

[0217] Table 13

[0218]

[0219] Table 14

[0220]

[0221]

[0222] * The sample was slurried at 5°C for 1 hour and turned into a black gel / oil.

[0223] All 14 hits were characterized by XRPD, TGA, DSC and solution NMR. The characterization results are summarized in Table 15.

[0224] Table 15

[0225]

[0226]

[0227] ND = Not Determined

[0228] Three salts were selected for further characterization - malate, fumarate and oxalate. The salts were scaled up to several hundred milligrams. The characterization data are summarized in Table 16.

[0229] Table 16

[0230]

[0231] *: Based on water adsorbed at 25°C / 80%RH: Very hygroscopic: >15%, Hygroscopic: 2-15%, Slightly hygroscopic: 0.2-2%, Non-hygroscopic: <0.2%.

[0232] Example 8: Additional polymorph screening of compound 1

[0233] Polymorph screening was performed using amorphous compound 1. To help design the experiment, the kinetic solubility of the compound was estimated. The estimation was performed using the solvent aliquot addition method, and the dissolution was judged by visual observation. The results are provided in Table 17. In Table 17, the solvent ratios (v / v) are approximate; the values ​​are rounded to the nearest integer. If complete dissolution is achieved by adding a single aliquot, the solubility is reported as ">"; if no solid is present, the solubility is reported as "<". The actual solubility may be greater than the calculated value due to the use of an excessively large solvent aliquot sample or due to a slow dissolution rate.

[0234] Table 17

[0235] Solvent system Solubility (mg / mL) ACN / water (60 / 40) <6 Dioxane / water (60 / 40) <13 Heptane >191 IPA 82 Methanol / water (40 / 60) <55* 2-MeTHF / MeOH / water (60 / 20 / 20) 106 NMP / water (68 / 32) 7

[0236] *Apparent hydrophobicity

[0237] Based on the solubility data, micro (about 5-10 mg) and medium (about 30-80 mg) scale crystallization experiments were designed using techniques such as slow evaporation of the melt, slurry and vapor effects. The addition of seed crystals and selected salt formers were also explored. The experiments included multiple steps, where observations from the initial steps guided the approach to subsequent steps.

[0238] The resulting samples were visually observed by polarized light microscopy and analyzed by XRPD for preliminary evaluation. If the resulting solid exhibited a unique XRPD pattern, the solution 1 They were further characterized by H NMR to confirm the chemical composition, and by TGA and DSC to evaluate the thermal behavior and the presence of volatiles. The screening conditions and results are summarized in Table 18.

[0239] Table 18

[0240]

[0241]

[0242]

[0243] BE = birefringence / extinction; SCXRD = single crystal X-ray diffraction

[0244] Table 19 provides a summary of the characterization data for the materials produced in this experiment. Refer to Table 15 for sample numbers.

[0245] Table 19

[0246]

[0247] Example 9: Polymorph Screening of Fumarate Salt of Compound 1

[0248] Compound 1 fumarate form A ethyl acetate solvate was used for polymorph screening. Form A ethyl acetate solvate was prepared as follows: amorphous compound 1 (3.0025 g) was suspended in ethyl acetate (60 mL) to obtain a clear solution. Fumaric acid (774.6 mg) was added to the solution and additional precipitation was observed. The mixture was stirred at ambient temperature for about a week. The formed solid was separated by filtration using an injector with positive displacement. Approximately 4.5 g of undried solid was recovered.

[0249] The screening consisted primarily of long-term slurry experiments. To aid in the design of the screening experiments, the kinetic solubility of the fumarate salt form A ethyl acetate solvate was estimated. The estimation was performed on a 3-11 mg scale using a solvent aliquot addition method, and the dissolution was judged by visual observation. The results are provided in Table 20. The solubility is estimated at ambient temperature and reported to the nearest mg / mL; if complete dissolution is achieved by a single aliquot addition, the solubility is reported as ">";

[0250] Table 20

[0251] Solvent system (ratio v%) Solubility (mg / mL) ACN <2* Anisole <1 t-BuOAc 1* 2-BuOH 17 DCE <9 DCE / EtOH(96 / 4) Dissolve at 8 Dioxane >104 IPA 20 IPA / Water(50 / 50) >124 2-MeTHF 32 MIBK 4 MTBE 2 TFE >100

[0252] *Reprecipitate after first aliquot.

[0253] Long-term slurry experiments were performed by stirring suspensions of Compound 1 fumarate Form A ethyl acetate solvate (about 30-100 mg) in various solvent systems at ambient temperature. The solvent system was selected based on solubility estimates. After stirring for 20 days, the solid was separated by centrifugal filtration (Table 21).

[0254] Table 21

[0255]

[0256]

[0257] Kinetic experiments using about 30-50 mg of Compound 1 fumarate form A ethyl acetate solvate include crystallization techniques such as fast and slow evaporation; solvent / antisolvent ripening precipitation; crystallization below ambient temperature; and organic and water vapor influencing factors (Table 22). For evaporation experiments, the filtered solution of the test material was left uncovered at ambient temperature for rapid evaporation, or covered with aluminum foil with pinholes for slow evaporation. For solvent / antisolvent precipitation, a solution of the starting material was prepared at ambient temperature or elevated temperature and filtered using a 0.2 μm nylon filter. The solution was mixed with an appropriate antisolvent by direct or reverse addition. The precipitated solid was either immediately separated by vacuum filtration or placed at ambient temperature for ripening. For crystallization below ambient temperature, a solution of the starting material was prepared at ambient temperature and filtered using a 0.2 μm nylon filter. The filtered solution was then placed under conditions below ambient temperature for slow crystallization. The precipitated solid was separated by centrifugal filtration. For the vapor effect experiment, the solid of the starting material was sampled into a vial, and the vial was placed in an RH tank at a set temperature (prepared as described in Greenspan, L., Journal of Research of the National Bureau of Standards Section A: Physics and Chemistry, Vol. 81A, No. 1, 1977, p. 89, doi: 10.6028 / jres.081a.011) or a secondary container filled with water. After a specified time, the sample was collected and analyzed.

[0258] Table 22

[0259]

[0260] BE = birefringence / extinction; SCXRD = single crystal X-ray diffraction

[0261] The glasses and films obtained in the kinetic experiments were crystallized by stirring for about 22 days (Table 23). The sample numbers refer to Table 22.

[0262] Table 23

[0263]

[0264] Table 24 provides a summary of the characterization data for the materials produced by this experiment.

[0265] Table 24

[0266]

[0267]

[0268]

[0269] Salt formation experiments were also performed by stirring a mixture of amorphous compound 1 and fumaric acid (1:1.2 ratio). The precipitated solid was separated by centrifugal filtration. The results are summarized in Table 25.

[0270] Table 25

[0271]

[0272]

Claims

1. A crystalline solid form of a fumarate salt of Compound 1: wherein the solid form is Form E.

2. The crystalline solid form of claim 1, wherein the crystalline solid form is characterized by one or more peaks in its XRPD pattern selected from those at about 5.83, about 7.03, about 8.69, about 12.88, about 13.43, about 14.68, about 15.65, about 16.65, and about 18.46 degrees 2θ.

3. The crystalline solid form of claim 1, wherein the crystalline solid form is characterized by peaks at about 5.83, about 7.03, about 8.69, about 12.88, about 13.43, about 14.68, about 15.65, about 16.65, and about 18.46 degrees 2θ in its XRPD pattern.

4. The crystalline solid form of claim 1, wherein the crystalline solid form is characterized by peaks in its XRPD pattern located at substantially all of the following positions:

5. The crystalline solid form of claim 1, wherein the crystalline solid form is characterized by one or more of the following: (i) an XRPD pattern substantially similar to the XRPD pattern depicted in Figure 1 and / or Figure 3; (ii) a TGA graph substantially similar to the TGA graph depicted in FIG. 2 and / or FIG. 4 ; and (iii) a DSC graph substantially similar to the DSC graph depicted in Figure 2 and / or Figure 5.

6. A pharmaceutical composition comprising the crystalline solid form of any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition of claim 6, wherein the pharmaceutical composition is solid.

8. The pharmaceutical composition of claim 6 or 7, wherein the pharmaceutical composition is formulated for oral administration.

9. A method of inhibiting an estrogen receptor or a mutant thereof in a biological sample, the method comprising contacting the biological sample with a crystalline solid form as claimed in any one of claims 1 to 5.

10. A method of inhibiting an estrogen receptor or a mutant thereof in a patient, the method comprising contacting the patient with a solid form as claimed in any one of claims 1 to 5.

11. A method of treating a disease, disorder or condition associated with an estrogen receptor in a patient, the method comprising administering to the patient a therapeutically effective amount of the crystalline solid form of any one of claims 1-5.

12. The method of claim 11, wherein the disease, disorder or condition is selected from the group consisting of breast cancer, bone cancer, lung cancer, colorectal cancer, endometrial cancer, prostate cancer, ovarian cancer, vaginal cancer, endometriosis and uterine cancer.

13. The method of claim 12, wherein the disease, disorder or condition is breast cancer.

14. The method of any one of claims 11-13, further comprising administering another anticancer agent.

15. The method of claim 14, wherein the anticancer agent is a CDK4 / 6 inhibitor, a PI3KCA inhibitor, or an mTOR inhibitor.

16. A method for preparing a crystalline solid form of a fumarate salt of Compound 1: Compound 1 Fumarate wherein the crystalline solid form is Form E, The method comprises making compound 1: Contact with fumaric acid in a suitable solvent to obtain Compound 1 fumarate salt Form E.

17. The method of claim 16, wherein the suitable solvent is isopropanol.

18. The method of claim 16 or 17, comprising dissolving Compound 1 in the suitable solvent and adding the fumaric acid portionwise.

19. The method of any one of claims 16-18, comprising adding seed crystals of Compound 1 fumarate Form E.

20. The process of claim 18 or 19, wherein at least one portion of fumaric acid is added prior to adding the seed crystals.

21. The method of any one of claims 18-20, wherein at least one portion of fumaric acid is added after adding the seed crystals.

22. A composition comprising a crystalline solid form of a fumarate salt of Compound 1: wherein the solid form is Form E, and wherein the composition is substantially free of impurities.

23. A composition comprising a crystalline solid form of a fumarate salt of Compound 1: wherein the solid form is Form E, and wherein the composition is substantially pure.

24. A composition comprising a crystalline solid form of a fumarate salt of Compound 1: wherein the solid form is Form E, and wherein the composition is substantially free of an amorphous solid form.

Images