Solid forms of 3-(5-fluorobenzofuran-3-yl)-4-(5-methyl-5h-[1,3]dioxolo[4,5-f]indol-7-yl)pyrrole-2,5-dione

Abstract

To provide novel solid forms of 3-(5-fluorobenzofuran-3-yl)-4-(5-methyl-5H-[1,3]dioxolo[4,5-f]indol-7-yl)pyrrole-2,5-dione, which is useful for the treatment of brain tumor, lung cancer, breast cancer, ovarian cancer, bladder cancer, neuroblastoma, kidney cancer, or pancreatic cancer.SOLUTION: The present invention provides a crystalline form II of 3-(5-fluorobenzofuran-3-yl)-4-(5-methyl-5H-[1,3]dioxolo[4,5-f]indol-7-yl)pyrrole-2,5-dione, characterized by an X-ray powder diffraction pattern.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 62 / 572,603, filed October 16, 2017, the entirety of which is incorporated herein by reference.

[0002] The present disclosure relates to solid forms of 3-(5-fluorobenzofuran-3-yl)-4-(5-methyl-5H-[1,3]dioxolo[4,5-f]indol-7-yl)pyrrole-2,5-dione, processes for its preparation, pharmaceutical compositions thereof, and uses thereof in the treatment of diseases. [Background technology]

[0003] 3-(5-fluorobenzofuran-3-yl)-4-(5-methyl-5H-[1,3]dioxolo[4,5-f]indol-7-yl)pyrrole-2,5-dione ("9-ING-41") has the following chemical structure: [ka]

[0004] 9-ING-41 is reported to be useful in the treatment of cancer, including brain, lung, breast, ovarian, bladder, neuroblastoma, kidney, and pancreatic cancer, as well as in the treatment of traumatic brain injury.

[0005] The structure, properties, and / or biological activity of 9-ING-41 are described in U.S. Pat. No. 8,207,216, Gaisina et al., From a Natural Product Lead to the Identification of Potent and Selective Benzofuran-3-yl-(indol-3-yl)maleimides as Glycogen Synthase Kinase 3β Inhibitors That Suppress Proliferation and Survival of Pancreatic Cancer Cells, J. Med. Chem. 2009, 52, 1853-1863, and Hilliard et al., Glycogen synthase kinase 3β inhibitors induce apoptosis in ovarian cancer cells and inhibit in-vivo tumor growth, Anti-Cancer Drugs 2011, 22:978-985.

[0006] There is a need for new solid forms of 9-ING-41, including polymorphs and solvates. Summary of the Invention

[0007] The present disclosure relates to solid forms of 9-ING-41, processes for preparing the solid forms of 9-ING-41, pharmaceutical compositions comprising the solid forms of 9-ING-41, and methods of treatment comprising administering the solid forms of 9-ING-41.

[0008] In some embodiments, the disclosure relates to a solid form that is crystalline Form II of 3-(5-fluorobenzofuran-3-yl)-4-(5-methyl-5H-[1,3]dioxolo[4,5-f]indol-7-yl)pyrrole-2,5-dione ("9-ING-41").

[0009] The present disclosure also provides a process for preparing solid forms of 9-ING-41.

[0010] The present disclosure also provides pharmaceutical compositions having solid forms of 9-ING-41.

[0011] The present disclosure also provides a method of treating a disease comprising administering to a patient in need thereof a therapeutically effective amount of a disclosed solid form of 9-ING-41. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 shows the X-ray powder diffraction diagram (XRPD) of Form II of 9-ING-41. [Diagram 2] FIG. 2 shows the X-ray powder diffraction diagram (XRPD) of Form II of 9-ING-41. [Diagram 3] FIG. 3 shows the differential scanning calorimetry (DSC) profile of Form II of 9-ING-41. [Figure 4] FIG. 4 shows the thermogravimetric analysis (TGA) profile of Form II of 9-ING-41. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] The present disclosure relates to solid state forms of 9-ING-41, processes for preparing same, and pharmaceutical compositions having the solid state forms. The present disclosure also relates to the conversion of the described solid state forms of 9-ING-41 to other solid state forms of 9-ING-41, 9-ING-41 salts, and those solid state forms.

[0014] The name "9-ING-41" is an alternative name for 3-(5-fluorobenzofuran-3-yl)-4-(5-methyl-5H-[1,3]dioxolo[4,5-f]indol-7-yl)pyrrole-2,5-dione, and these names are used interchangeably herein.

[0015] The solid forms of 9-ING-41 according to the present disclosure may have advantageous properties selected from at least one of chemical or polymorphic purity, flowability, solubility, dissolution rate, bioavailability, morphology or crystal habit, chemical stability with respect to polymorphic transformation, thermal and mechanical stability, stability such as stability with respect to dehydration and / or storage stability, low hygroscopicity, low residual solvent content, and advantageous processing and handling properties such as compressibility and bulk density.

[0016] Herein, a crystalline form may be referred to as being characterized by graphical data as "shown" in a figure. Such data may include, for example, X-ray powder diffractograms (XRPD), differential scanning calorimetry (DSC) thermograms, thermogravimetric analysis (TGA) profiles, and dynamic vapor sorption profiles (DVS). As is well known in the art, graphical data may provide additional technical information that further defines each solid form that cannot necessarily be described by reference to numerical values ​​or peak positions alone. Thus, the term "substantially shown" when referring to the graphical data in the figures herein is not necessarily identical to that depicted herein, but falls within the experimental error or deviation as considered by those skilled in the art. Those skilled in the art will be able to easily compare the graphical data in the figures herein with graphical data generated for an unknown crystalline form and ascertain whether the two sets of graphical data characterize the same crystalline form or two different crystalline forms.

[0017] The solid, crystalline forms may be referred to herein as "polymorphically pure" or "substantially free of other forms." In this context, the phrase "substantially free of other forms" is understood to mean that the solid form contains about 20% or less, about 10% or less, about 5% or less, about 2% or less, about 1% or less, or 0% of other forms of the subject compound as measured, for example, by XRPD. Thus, solid forms of 9-ING-41 described herein as being substantially free of other solid forms are understood to contain about 80% (w / w) or more, about 90% (w / w) or more, about 95% (w / w) or more, about 98% (w / w) or more, about 99% (w / w) or more, or about 100% of the subject solid form of 9-ING-41. Thus, in some embodiments of the present disclosure, the described solid forms of 9-ING-41 may contain about 1% to about 20% (w / w), about 5% to about 20% (w / w), or about 5% to about 10% (w / w) of one or more other solid forms of 9-ING-41.

[0018] As used herein, unless otherwise stated, XRPD peaks reported herein are measured using CuKα radiation, λ=1.5419 Å.

[0019] The modifier "about" should be considered to disclose a range defined by the absolute values ​​of the two endpoints. For example, the phrase "about 2 to about 4" also discloses a range of "2 to 4." When used to modify a single number, the term "about" refers to plus or minus 10% of the indicated number and includes the indicated number. For example, "about 10%" indicates a range of 9% to 11%, and "about 1" means 0.9 to 1.1.

[0020] The term "solvate" as used herein refers to a crystalline form that incorporates a solvent into the crystalline structure, unless otherwise specified. When the solvent is water, the solvate is often called a "hydrate". The solvent in the solvate may be present in stoichiometric or non-stoichiometric amounts.

[0021] In some aspects, the present disclosure relates to solid forms of 9-ING-41.

[0022] In some embodiments, the solid form of 9-ING-41 is crystalline Form II of 9-ING-41. In other embodiments, the solid form is crystalline Form II of 9-ING-41 substantially free of other solid forms of 9-ING-41. Crystalline Form II of 9-ING-41 exhibits an XRPD substantially as shown in FIG.

[0023] The XRPD of crystalline Form II of 9-ING-41 shown in FIG. 1 has reflection angles (degrees 2-theta ±0.2 degrees 2-theta) as shown in Table 1. [Table 1] TIFF2024001040000004.tif197158

[0024] In some embodiments of the present disclosure, crystalline form II of 9-ING-41 is characterized by an XRPD pattern having a peak at one of the angles listed in Table 1. In other embodiments, crystalline form II of 9-ING-41 is characterized by an XRPD pattern having a plurality of peaks at one of the angles listed in Table 1 above. In other embodiments, crystalline form II of 9-ING-41 is characterized by an XRPD pattern having two peaks selected from the angles listed in Table 1 above. In other embodiments, crystalline form II of 9-ING-41 is characterized by an XRPD pattern including three peaks selected from the angles listed in Table 1 above. In other embodiments, crystalline form II of 9-ING-41 is characterized by an XRPD pattern including four peaks selected from the angles listed in Table 1 above. In other embodiments, crystalline form I of 9-ING-41 is characterized by an XRPD pattern including five peaks selected from the angles listed in Table 1 above. In another embodiment, crystalline form II of 9-ING-41 is characterized by an XRPD pattern that includes six peaks selected from the angles listed in Table 1 above. In another embodiment, crystalline form II of 9-ING-41 is characterized by an XRPD pattern that includes seven peaks selected from the angles listed in Table 1 above. In another embodiment, crystalline form II of 9-ING-41 is characterized by an XRPD pattern that includes eight peaks selected from the angles listed in Table 1 above. In another embodiment, crystalline form II of 9-ING-41 is characterized by an XRPD pattern that includes nine peaks selected from the angles listed in Table 1 above. In another embodiment, crystalline form II of 9-ING-41 is characterized by an XRPD pattern that includes ten peaks selected from the angles listed in Table 1 above. In another embodiment, crystalline form II of 9-ING-41 is characterized by an XRPD pattern that includes more than ten peaks selected from the angles listed in Table 1 above.

[0025] In some embodiments, crystalline form II of 9-ING-41 is characterized by an XRPD pattern with peaks at 7.8 degrees ± 0.2 degrees 2-theta. In other embodiments, crystalline form II of 9-ING-41 is characterized by an XRPD pattern with peaks at 7.8, 12.9, 15.8, and 18.9 degrees ± 0.2 degrees 2-theta. In other embodiments, crystalline form II of 9-ING-41 is characterized by an XRPD pattern with peaks at 12.9, 18.1, 28.0, and 28.5 degrees ± 0.2 degrees 2-theta.

[0026] In some embodiments of the present disclosure, crystalline Form II of 9-ING-41 is characterized by an XRPD pattern with peaks at three or more of the following: 7.8, 12.9, 15.8, 18.1, 18.9, 19.5, 22.1, 22.7, 28.0, and 28.5 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, crystalline Form II of 9-ING-41 is characterized by an XRPD pattern with peaks at four or more of the following: 7.8, 12.9, 15.8, 18.1, 18.9, 19.5, 22.1, 22.7, 28.0, and 28.5 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, crystalline Form II of 9-ING-41 is characterized by an XRPD pattern with peaks at five or more of the following: 7.8, 12.9, 15.8, 18.1, 18.9, 19.5, 22.1, 22.7, 28.0, and 28.5 degrees ± 0.2 degrees 2-theta. In some embodiments of the present disclosure, crystalline Form II of 9-ING-41 is characterized by an XRPD pattern with peaks at six or more of the following: 7.8, 12.9, 15.8, 18.1, 18.9, 19.5, 22.1, 22.7, 28.0, and 28.5 degrees ± 0.2 degrees 2-theta. It is characterized by an XRPD pattern having no less than seven peaks at 7.8, 12.9, 15.8, 18.1, 18.9, 19.5, 22.1, 22.7, 28.0 and 28.5 degrees ±0.2 degrees two-theta.

[0027] In some embodiments, crystalline Form II of 9-ING-41 exhibits an XRPD pattern substantially as shown in FIG.

[0028] Crystalline Form II of 9-ING-41 can be characterized by a DSC thermogram substantially as shown in Figure 3. As shown in Figure 3, crystalline Form II of 9-ING-41 produces an endothermic peak at 218.03°C with a peak onset temperature of 216.14°C and a melting enthalpy of 53.441 J / g when heated at a rate of 10°C / min. In some embodiments of the present disclosure, crystalline Form II of 9-ING-41 is characterized by a DSC thermogram with an endothermic peak at about 218°C. In other embodiments of the present disclosure, crystalline Form II of 9-ING-41 is characterized by a DSC melting enthalpy of about 53 J / g.

[0029] Crystalline Form II of 9-ING-41, when heated at a rate of 10° C. / min, may be characterized by a TGA profile substantially as shown in Figure 4. As shown in Figure 4, crystalline Form II of 9-ING-41 lost about 0.3521% of its weight upon heating between about 150° C. and about 250° C. when heated at a rate of 10° C. / min.

[0030] In some embodiments of the present disclosure, crystalline Form II of 9-ING-41 is characterized by an XRPD pattern with peaks at 7.8, 12.9, 15.8, 18.1, 18.9, 19.5, 22.1, 22.7, 28.0 and 28.5 degrees ± 0.2 degrees 2-theta, and a DSC thermogram with an endothermic peak at about 218°C when heated at a rate of 10°C / min.

[0031] In some embodiments, the present disclosure relates to a process for preparing the disclosed solid forms of 9-ING-41, hi some embodiments, the process comprises concentrating a solution of 9-ING-41 dissolved in a solvent or solvent mixture.

[0032] In some aspects, the disclosure relates to a process for preparing crystalline Form II of 9-ING-41. In some embodiments, the process comprises concentrating (e.g., in a vacuum or by evaporation) a solution of 9-ING-41 in a mixture of 3-methylbutanol / acetone in a volume ratio of about 1:1. In other embodiments, the process comprises cooling a warm (i.e., above 25° C.) solution of 9-ING-41 in a mixture of isoamyl alcohol:acetonitrile in a volume ratio of about 10:1.

[0033] In other aspects, the disclosure encompasses pharmaceutical compositions having a solid form of 9-ING-41 of the disclosure and at least one pharma- ceutically acceptable excipient. Pharmaceutically acceptable excipients will be known to those skilled in the art. The pharmaceutical composition can be administered in any convenient dosage form. Exemplary dosage forms include tablets, capsules, caplets, reconstitutable powders, elixirs, liquids, colloidal or other types of suspensions, emulsions, beads, beadlets, granules, microparticles, nanoparticles, and combinations thereof. The amount of the composition administered will depend on the subject being treated, the subject's weight, the severity of the condition being treated, the method of administration, and the judgment of the prescribing physician. In some embodiments, the pharmaceutical composition has crystalline Form II of 9-ING-41 and at least one pharma-ceutically acceptable excipient.

[0034] The present disclosure also encompasses sterile aqueous or organic solution formulations of 9-ING-41, the formulations being prepared from the solid forms of 9-ING-41 of the present disclosure. Thus, in some aspects, the present disclosure includes a process for preparing a pharmaceutical composition that is a solution having 9-ING-41. In some embodiments, the method for preparing a pharmaceutical composition that is a solution of 9-ING-41 includes dissolving a solid form of 9-ING-41 of the present disclosure in a solvent or solvent mixture. In some embodiments, the method includes dissolving crystalline Form II of 9-ING-41 in an aqueous solvent, a non-aqueous solvent, or a mixture of aqueous and / or non-aqueous solvents. The aqueous solvent, non-aqueous solvent, or mixture of aqueous and / or non-aqueous solvents in embodiments can include other dissolving components, such as, for example, polyethylene glycol, benzyl alcohol, polysorbate, tocopheryl polyethylene glycol succinate, as well as other surfactants, solubilizers, or other pharma- ceutically acceptable excipients. In another embodiment, the process comprises dissolving crystalline Form II of 9-ING-41 in an aqueous solvent.

[0035] The solid state forms of 9-ING-41 defined herein, as well as pharmaceutical compositions or formulations thereof, can be used as medicaments for the treatment of cancer, including, inter alia, brain tumors, lung cancer, breast cancer, ovarian cancer, bladder cancer, neuroblastoma, kidney cancer, and pancreatic cancer, as well as the treatment of traumatic brain injury.

[0036] Although the present disclosure has been described with reference to certain preferred embodiments, other embodiments will become apparent to those skilled in the art from consideration of the specification. The present disclosure is further illustrated by reference to the following examples. It will be apparent to those skilled in the art that many modifications, both to materials and methods, can be practiced without departing from the scope of the present disclosure.

[0037] Analysis method XRPD analysis XRPD analysis was carried out using an X-ray diffractometer (Bruker D8 Advance) equipped with a LynxEye detector, with the following instrumental parameters: Scan: 3°(2θ)~40°(2θ) Increment: 0.02°(2θ) Scan speed: 0.3 seconds / step Voltage: 40KV Current: 40mA Rotation: On Sample Hold: Zero background sample holder

[0038] The samples were scanned from 3° to 40° (2θ) with a step of 0.02° (2θ) on a D2 phaser X-ray powder diffractometer (Bruker). The tube voltage and current were 30 KV and 10 mA, respectively.

[0039] TGA analysis TGA analysis was performed on a TA Instruments TGA Q500 or a Discovery TGA 55 (TA Instruments, US). Samples were placed in tarred open aluminum pans and heated at a rate of 10 °C / min from room temperature to the final temperature.

[0040] DSC analysis DSC analysis was performed on a DSC Q200 or Discovery DSC 250 (TA Instruments, US). A weighted sample was placed in the DSC pinhole pan and the weight was accurately recorded. The sample was heated at a rate of 10°C / min to the final temperature.

[0041] DVS analysis DVS analysis was performed on an IGAsorp (HidenIsochema Ltd.) For isothermal studies, the chamber temperature was maintained at a constant 25.0 ± 1.0 °C water bath.

[0042] Polarized Light Microscope (PLM) PLM analysis was performed using a polarizing microscope ECLIPSE LV100POL (Nikon, JPN).

[0043] HPLC analysis Equipment: Agilent 1260 Infinity Series Diluent: Acetonitrile Flow rate: 1.5mL / min Mobile phase: A: 0.05% TFA in water B: 0.05% TFA in acetonitrile Injection volume: 1μL Column: XDB-C18, 4.6*50mm, 1.8μm Column temperature: 40℃ Detection: 220nm Run Time: 8 minutes (2 minute delay before next injection) Gradient (T / B%): 0.0 / 30, 6.0 / 100, 8.0 / 100

[0044] Working Example Example 1: Preparation of 9-ING-41 Crude 9-ING-41 can be obtained by the general methods described in U.S. Pat. No. 8,207,216 and Gaisina et al., From a Natural Product Lead to the Identification of Potent and Selective Benzofuran-3-yl-(indol-3-yl)maleimides as Glycogen Synthase Kinase 3β Inhibitors That Suppress Proliferation and Survival of Pancreatic Cancer Cells, J. Med. Chem. 2009, 52, 1853-1863).

[0045] Example 2: Preparation of 9-ING-41 Crystalline Form I Crystalline Form I of 9-ING-41 can be prepared by: Synthesis of intermediate 1 [ka]

[0046] A 3 L four-neck round bottom flask purged and maintained with an inert atmosphere of nitrogen was charged with 6-nitro-2H-1,3-benzodioxole-5-carbaldehyde (200 g, 1.02 mol, 1.00 equiv.), ammonium acetate (200 g, 2.59 mol, 2.53 equiv.), acetic acid (2 L), and nitromethane (313 g, 5.13 mol, 5.00 equiv.). The solution was stirred at 100° C. for 12 hours. The reaction was repeated three times. The solutions were combined and diluted with 20 L of water. The resulting solution was extracted with 3×10 L of ethyl acetate and the organic layers were combined. The mixture was washed with 3×10 L of brine, dried over anhydrous sodium sulfate, and concentrated under vacuum. This gave 450 g (crude) of 5-nitro-6-[(E)-2-nitroethenyl]-2H-1,3-benzodioxole (1) as a dark green solid. Synthesis of intermediate 2 [ka]

[0047] Iron (120 g, 2.14 mol, 17.01 equiv.) was slowly added to a suspension of 5-nitro-6-[(Z)-2-nitroethenyl]-2H-1,3-benzodioxole (30 g, 125.97 mmol, 1.00 equiv.), silica gel (120 g), toluene (200 mL), and cyclohexane (400 mL) in acetic acid (300 mL) at 80° C. under nitrogen. The resulting black mixture was stirred at 80° C. for 8 h. The reaction was repeated 10 times. The reaction mixture was combined. The solid was filtered off. The filtrate was concentrated under vacuum and the residue was applied to a silica gel column with ethyl acetate / petroleum ether (1 / 5). The collected fractions were combined and concentrated under vacuum to give 67.3 g (33%) of 2H,5H-[1,3]dioxolo[4,5-f]indole (2) as an off-white solid. Synthesis of intermediate 3 [ka]

[0048] Sodium hydride (19.9 g, 497.50 mmol, 1.18 equiv, 60%) was added portionwise to a solution of 2H,3H,5H-furo[2,3-f]indole (67.3 g, 422.78 mmol, 1.00 equiv) in N,N-dimethylformamide (1.3 L) at 0° C. under nitrogen. The mixture was stirred at 0° C. for 1 h and CHI (70.9 g, 499.51 mmol, 1.18 equiv) was added dropwise. The resulting solution was stirred at room temperature for 3 h. The solution was quenched by adding 1 L of ice water. The resulting solution was extracted with 3×1 L of ethyl acetate and the organic layers were combined. The mixture was washed with 3×1 L of brine, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was loaded onto a silica gel column with ethyl acetate / petroleum ether (1 / 10). The collected fractions were combined and concentrated in vacuo to give 71 g (97%) of 5-methyl-2H,3H,5H-furo[2,3-f]indole (3) as a pale yellow solid. Synthesis of intermediate 4 [ka]

[0049] Ethyl 2-chloro-2-oxoacetate (220 g, 1.61 mol, 3.96 equiv.) was added dropwise to a solution of 5-methyl-2H,3H,5H-furo[2,3-f]indole (70.4 g, 406.44 mmol, 1.00 equiv.) in ethyl ether (1.6 L) at 0 °C under nitrogen. The resulting solution was allowed to warm to room temperature and stirred for 4 h. The reaction was slowly quenched by adding 2 L of ice water and the pH value of the resulting solution was adjusted to 9 with Na2CO3. The resulting mixture was extracted with 3 × 1.5 L of ethyl acetate. The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under vacuum to give 92.8 g (84%) of ethyl 2-[5-methyl-2H,3H,5H-furo[2,3-f]indol-7-yl]-2-oxoacetate (4) as a pale yellow solid.

[0050] 1H NMR (300 MHz, DMSO-d6): δ 8.28(s,4H),7.56(s,4H),7.27(s,4H),6.17(s,1H),6.08(s,8H),4.35(q,J=7.1 Hz,7H),3.85(s,11H),3.35(s,2H),1.35(t,J=7.1 Hz,11H),1.25(s,2H). Synthesis of intermediate 5 [ka]

[0051] A 10 L four-neck round bottom flask was charged with 2-bromo-4-fluorophenol (500 g, 2.62 mol, 1.00 equiv), N,N-dimethylformamide (5 L), potassium carbonate (1253 g, 9.07 mol, 3.46 equiv) and ethyl (2E)-4-bromobut-2-enoate (1010 g, 5.23 mol, 2.00 equiv). The resulting solution was stirred at room temperature for 12 h. The solid was collected by filtration. The reaction was then quenched by adding 15 L of water and extracted with 3×10 L of ethyl acetate. The organic layers were combined and washed with 4×20 L of brine. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was loaded onto a silica gel column with ethyl acetate / petroleum ether (1 / 20). The collected fractions were combined and concentrated in vacuo to give 500 g (63%) of ethyl (2E)-4-(2-bromo-4-fluorophenoxy)but-2-enoate (5) as a white solid. Synthesis of intermediate 6 [ka]

[0052] A 2 L three-neck round bottom flask purged and maintained under an inert atmosphere of nitrogen was charged with ethyl (2E)-4-(2-bromo-4-fluorophenoxy)but-2-enoate (125 g, 412.37 mmol, 1.00 equiv), benzyltriethylazanium chloride (99 g, 434.64 mmol, 1.05 equiv), sodium formate dihydrate (45.1 g), Pd(OAc)2 (2.9 g, 12.92 mmol, 0.03 equiv), sodium carbonate (92 g, 868.01 mmol, 2.10 equiv), and N,N-dimethylformamide (1.25 L). The resulting solution was stirred at 80° C. for 12 hours. The reaction was repeated four times. The reaction mixture was combined and the solids were filtered off. The filtrate was diluted with 10 L of brine and extracted with 3×5 L of ethyl acetate. The organic layers were combined and washed with 4×6 L of brine. The mixture was dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was applied to a silica gel column with ethyl acetate / petroleum ether (1 / 20). The collected fractions were combined and concentrated under vacuum. This gave 258 g (crude) of ethyl 2-(5-fluoro-1-benzofuran-3-yl)acetate (6) as a pale yellow oil. Synthesis of intermediate 7 [ka]

[0053] A 5 L round bottom flask was charged with ethyl 2-(5-fluoro-1-benzofuran-3-yl)acetate (147 g, 661.53 mmol, 1.00 equiv), methanol (1 L), tetrahydrofuran (1 L), water (1 L), and LiOH (47.7 g, 1.99 mol, 3.01 equiv). The resulting solution was stirred at room temperature for 3 h. The reaction was repeated twice. The mixture was concentrated under vacuum and then extracted with 1 L of dichloromethane. The aqueous layer was collected and the pH of the layer was adjusted to 1-3 with hydrogen chloride (1 mol / L). The resulting solution was extracted with 3×1 L of ethyl acetate and the combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum. This afforded 160 g (62%) of 2-(5-fluoro-1-benzofuran-3-yl)acetic acid (7) as a white solid. Synthesis of intermediate 8 [ka]

[0054] A 10 L round bottom flask was charged with 2-(5-fluoro-1-benzofuran-3-yl)acetic acid (160 g, 824.1 mmol, 1.00 equiv), NH4Cl (436 g, 8.16 mol, 9.89 equiv), N,N-dimethylformamide (6 L), DIEA (1064 g, 8.24 mol, 9.99 equiv), and HATU (376 g, 988.88 mmol, 1.20 equiv). The resulting solution was stirred at room temperature for 12 h. The resulting solution was diluted with 10 L of water. The solid was collected by filtration to give 126 g (78%) of 2-(5-fluoro-1-benzofuran-3-yl)acetamide (8) as a white solid. Synthesis of 9-ING-41 in crystalline form I [ka]

[0055] t-BuOK (1200 mL, 1 mol / L in THF) was added dropwise to a solution of ethyl 2-[5-methyl-2H,3H,5H-furo[2,3-f]indol-7-yl]-2-oxoacetate (100 g, 365.9 mmol, 1.00 equiv.), 2-(5-fluoro-1-benzofuran-3-yl)acetamide (72 g, 372.7 mmol, 1.02 equiv.), tetrahydrofuran (3 L) at 0° C. under nitrogen. The reaction was stirred at room temperature for 2 h. The reaction was cooled to 0° C., poured into 2 L of NH4Cl (saturated solution in water) and extracted with 4×2 L of dichloromethane. The organic layers were combined, dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was loaded onto a silica gel column with ethyl acetate / dichloromethane / petroleum ether (1 / 1 / 5). The collected fractions were combined and concentrated under vacuum to give 107.9 g (74%) of 3-(5-fluoro-1-benzofuran-3-yl)-4-[5-methyl-2H,5H-[1,3]dioxolo[4,5-f]indol-7-yl]-2,5-dihydro-1H-pyrrole-2,5-dione as a red solid. The red solid is 9-ING-41 crystalline form I. MS-ESI: [M+H]+=405.

[0056] Example 3: Preparation of 9-ING-41 Crystalline Form II Crystalline Form II of 9-ING-41 was prepared by slow evaporation of a solution of 9-ING-41 as follows: 29.12 mg of 9-ING-41 (Form I) was weighed into a glass vial and 3 mL of 3-methylbutanol was added. 80.43 mg of 9-ING-41 (Form I) was weighed into another glass vial and 3 mL of acetone was added. The resulting suspension was filtered and 100 μL of each filtrate was mixed in a well of a 96-well plate. The mixture was allowed to evaporate under ambient conditions in a laboratory fume hood to give solid 9-ING-41 crystalline Form II. This material produced the XRPD shown in Figure 1.

[0057] Example 4: Preparation of 9-ING-41 Crystalline Form II Approximately 100 mg of 9-ING-41 Form I was weighed into a vial. 7 ml of a 10:1 (vol / vol) mixture of isoamyl alcohol:acetonitrile was added to the vial. The mixture was heated at 80° C. to obtain a solution which was subsequently filtered and seeded with Form II 9-ING-41. The mixture was stirred while cooling to room temperature and crystallized. XRPD showed the resulting solid to be Form II.

[0058] Example 5: Preparation of 9-ING-41 Crystalline Form II Approximately 2.0 g of Form I was weighed into a vial. 150 ml of a 10:1 (vol / vol) mixture of isoamyl alcohol:acetonitrile was added to the vial. The mixture was heated at 90° C. to obtain a solution which was subsequently filtered and seeded with Form II 9-ING-41. The mixture was stirred while cooling to room temperature and crystallized. The crystalline solid (1.46 g) was characterized by PLM, XRPD, DSC, and TGA. PLM showed the crystalline solid to be irregular clumps of crystals. XRPD (see FIG. 2) showed the crystalline solid to be Form II. DSC showed a first endotherm with an onset temperature of 216° C. attributed to melting of Form II, followed by an exotherm attributed to Form I crystallization, followed by a second endotherm with an onset temperature of 228° C. (see FIG. 3). This is attributed to melting of Form I. TGA showed the form to decrease by approximately 0.4% by weight before 250° C. See FIG. 4.

[0059] Example 6: Mechanical and Pressure Effects 10.0 mg of 9-ING-41 Form II was weighed into a mortar and ground for 5 minutes. The remaining solid was collected and analyzed by XRPD. The ground solid remained as Form II 9-ING-41.

[0060] 4.50 mg of Form II was compressed into a tablet. The tablet was then ground into a powder and analyzed by XRPD. Compression did not affect the crystalline morphology.

[0061] Example 7: Heat Treatment - Preparation of Amorphous 9-ING-41 9-ING-41 Form II was heated by DSC at a rate of 10° C. / min to a final temperature of 260° C. The molten product was immediately placed in an ice bath and held for 15 minutes. The residue was analyzed by XRPD, which showed the material to be amorphous.

[0062] 9-ING-41 Form II was heated at a rate of 10° C. / min to a final temperature of 260° C., then cooled to −40° C., reheated to 260° C. at the same heating rate, then cooled to 40° C. Analysis of the residue by XRPD showed the material to be amorphous.

[0063] Example 8: Solubility Study - 24 hours 9-ING-41, either Form I or Form II, was added to 1 mL of each of 10 different solvents in a vial to create a suspension. The samples were shaken at 2000 rpm in a shaker at room temperature for 24 hours. The mixtures were visually inspected to confirm saturation. After 24 hours, 1 mL of the suspension was filtered through a 0.45 μm nylon microfiltration membrane into separate clear glass vials. The filtrates were analyzed by HPLC and the remaining solids were analyzed using XRPD. The solubility results are shown in Table 2 below. [Table 2]

[0064] The remaining solids in experiments in which Form I was the starting material remained as Form I.

[0065] The remaining solid in the experiments where Form II was the starting material changed form, except for the water and sesame oil experiments, where the remaining solid was Form II. In the ethanol experiment, the remaining solid was a solvate. In the other experiments, the solid appeared to be Form I.

[0066] Example 9: Solubility Study - 1 Hour Step 1: A sufficient amount of Form I was added to 1 mL of solvent (EtOH, Labrasol, benzyl benzoate, benzyl alcohol, PEG400:Tween80:EtOH (75:8:17) and PEG300:BzOH:water (65:10:25)) to make a suspension. The sample was shaken at 2000 rpm in a shaker at room temperature for 24 hours.

[0067] Step 2: The vials were visually inspected to ensure saturation. 1 mL of the suspension was filtered through a 0.45 μm nylon microfiltration membrane into another clean, clear glass vial for 24 hours. The filtrate was analyzed by HPLC.

[0068] Step 3: Excess Form II was added to each of the six remaining filtrates from step 2 to create a suspension. The samples were shaken at 2000 rpm on a shaker for 1 hour. After 1 hour, the suspension was filtered through a 0.45 μm nylon microfiltration membrane into another clean glass vial. The filtrate was analyzed by HPLC and the remaining solid was analyzed using XRPD. [Table 3]

[0069] The remaining solid in Labrasol, benzyl benzoate, benzyl alcohol, and PEG400:Tween80:EtOH (75:8:17) was Form I, indicating that Form II of the solid rapidly converted to Form I. The crystalline form of the remaining solid did not change in EtOH and PEG300:BzOH:water (65:10:25).

[0070] Example 10: Solubility in Sesame Oil The solubility of 9-ING-41 in sesame oil cannot be measured by HPLC. Solubility was estimated by adding weighed portions of Form I (or Form II) to sesame oil. Samples were stirred at room temperature for 24 hours and dissolution was confirmed by visually inspecting the vials. Solids were added in this manner until no added solids dissolved in 24 hours. The results are shown in Table 4 below. [Table 4]

Claims

1. 1. An amorphous solid of 3-(5-fluorobenzofuran-3-yl)-4-(5-methyl-5H-[1,3]dioxolo[4,5-f]indol-7-yl)pyrrole-2,5-dione (9-ING-41), comprising: (a) heating crystalline Form II of 9-ING-41 to 260°C at a rate of 10°C / min, followed by holding at 0°C for 15 minutes; or (b) heating crystalline Form II of 9-ING-41 to 260°C at a rate of 10°C / min, then cooling to -40°C, then reheating to 260°C at a rate of 10°C / min, and then cooling to 40°C. and The crystalline Form II of 9-ING-41 is characterized by an X-ray powder diffraction pattern with peaks at 7.8, 12.9, 15.8, 18.1, 18.9, 28.0, and 28.5 degrees ±0.2 degrees two-theta on the 2-theta scale at lambda=1.54 Angstroms (Cu Kα), and a differential scanning calorimetry (DSC) thermogram with an endothermic peak at about 218° C. when heated at a rate of 10° C. / min. Amorphous solid.

2. A pharmaceutical composition comprising the amorphous solid of claim 1 and a pharmaceutically acceptable excipient.

3. Use of a therapeutically effective amount of the amorphous solid of claim 1 in the manufacture of a medicament for treating cancer.

4. The use of claim 3, wherein the cancer is brain tumor, lung cancer, breast cancer, ovarian cancer, bladder cancer, neuroblastoma, kidney cancer, or pancreatic cancer.

5. Use of a therapeutically effective amount of the amorphous solid of claim 1 in the manufacture of a medicament for treating traumatic brain injury in a patient.

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