Crystal complex

Abstract

The present invention relates to a crystalline complex of sulforaphane and α-cyclodextrin, pharmaceutical compositions comprising the complex; methods for their preparation; and the use of said complex as a medicament.

Description

[Technical Field]

[0001] The present invention relates to a crystalline complex of sulforaphane and α-cyclodextrin, pharmaceutical compositions comprising the complex; methods for their preparation; and the use of said complex as a medicament. [Background technology]

[0002] Sulforaphane is a compound derived from cruciferous vegetables such as cabbage, broccoli, broccoli sprouts, Brussels sprouts, cauliflower, cauliflower sprouts, bok choy, kale, collard greens, arugula, kohlrabi, mustard, turnip, red radish, and watercress. In plants, it exists in a bound form as the glucosinolate glucoraphanin. In nature, sulforaphane is formed from glucoraphanin by the enzyme myrosinase following damage to plant cells caused by, for example, chewing.

[0003] Racemic sulforaphane, also known as 4-methylsulfinylbutyl isothiocyanate, has the following structure: [ka]

[0004] Naturally occurring sulforaphane is chiral and exists predominantly in the (R) configuration.

[0005] Sulforaphane has recently been shown to upregulate glutathione, leading to antioxidant and detoxifying properties [Zhang & Tang, Acta Pharm. Sinica (2007), 1343-1354; Pastore et al., Clinica Chimica Acta (2003), 19-39]; activate nuclear factor erythroid 2-related factor 2 (Nrf2), leading to anti-inflammatory and anti-proliferative effects [Houghton, Oxidative Medicine & Cellular Longevity (2019), https: / / doi.org / 10.1155 / 2019 / 2716870]; and inactivate signal transducer and activator of transcription 3 (STAT3), leading to anti-angiogenic and apoptotic properties relevant to oncology [Liu et al., Nature Scientific Reports (2017), doi:10.1038 / s41598-017-12855-w; Clarke et al., Cancer Letters(2008), 291-304; Lenzi et al., Cancer Treat. Res.(2014), 207-223]; and binding to macrophage inhibitory factor (MIF) and nuclear factor kappa-light-chain-enhancer of activated B cells [Clulow et al., Chem. Comm.(2017), doi:10.1039 / c6cc08797c].

[0006] Despite sulforaphane's significant therapeutic potential, its development as a pharmaceutical has been hindered by the compound's inherent chemical instability. Sulforaphane exists in the form of an unstable oil that rapidly decomposes under ambient conditions. This makes sulforaphane particularly difficult to manufacture, formulate, and distribute.

[0007] Complexation of sulforaphane with cyclodextrin has been shown to be a particularly effective approach for stabilizing sulforaphane. International Publication No. 2008 / 091608 describes the synthesis of racemic sulforaphane and the preparation of stabilized complexes of sulforaphane with α-, β-, and γ-cyclodextrin. The Examples section therein describes different methods for preparing specific α-cyclodextrin complexes, with the highest reported sulforaphane loading being 7.1 wt %, corresponding to a molar ratio of sulforaphane to α-cyclodextrin of approximately 1:2. International Publication No. 2013 / 179057 describes an improved scale-up procedure for the synthesis of sulforaphane and the preparation of a complex of sulforaphane with α-cyclodextrin. WO 2013 / 179056 discloses a method for preparing a sulforaphane:cyclodextrin complex by isolating chiral (R)-sulforaphane from a crude natural extract via cyclodextrin complexation. Summary of the Invention [Problem to be solved by the invention]

[0008] A robust, reproducible, and stable solid state of stabilized sulforaphane is required to be suitable for large-scale commercial production, handling, and storage.

[0009] The present invention has been devised with the above in mind. [Means for solving the problem]

[0010] Disclosed herein is a crystalline complex of sulforaphane and α-cyclodextrin that has low water content and improved stability over a range of ambient humidity conditions. The complex may have long-term stability at temperatures up to 40° C. The complex may have long-term stability at up to 70% relative humidity.

[0011] According to a first aspect of the present invention, there is provided a crystalline complex of sulforaphane and α-cyclodextrin having a water content of less than 8% w / w, wherein the crystalline form of the complex is stable at 25°C and between 0% and 60% relative humidity (RH).

[0012] According to a second aspect, there is provided a crystalline complex of sulforaphane and α-cyclodextrin (Form 3), wherein the crystalline form of the complex is characterized by X-ray powder diffraction (XRPD) peaks at 5.3 and 10.7±0.2 degrees 2θ, when measured in reflectance mode.

[0013] According to a third aspect of the present invention, there is provided a method for forming a crystalline complex of sulforaphane and α-cyclodextrin according to the first or second aspect, the method comprising: a) providing a complex of sulforaphane and α-cyclodextrin; b) drying the composite from step a) while stirring the composite at a pressure of less than 200 mbar until the moisture content of the composite is less than 6% w / w; The compound comprises:

[0014] According to a fourth aspect of the present invention, there is provided a solid pharmaceutical composition comprising an effective amount of a crystalline complex according to the first or second aspect of the present invention, and optionally at least one pharmaceutically acceptable excipient.

[0015] According to a fifth aspect of the present invention, there is provided a crystalline complex according to the first or second aspect of the present invention, or a pharmaceutical composition according to the fourth aspect of the present invention, for use in therapy. In one embodiment, the crystalline complex or pharmaceutical composition is used to treat a disease or disorder mediated by Nrf2 or STAT3. In one embodiment, the crystalline complex or pharmaceutical composition is used to treat a disease or disorder mediated by Nrf2 or STAT3, including cancer, subarachnoid hemorrhage, intracerebral hemorrhage, ischemic stroke, delayed cerebral ischemia, atherosclerosis, middle cerebral artery infarction, pulmonary arterial hypertension, Alport syndrome, non-alcoholic steatohepatitis, focal segmental glomerulosclerosis, Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, cirrhosis, non-alcoholic fatty liver disease, hepatitis, acute kidney injury, sepsis, chronic kidney disease, systemic lupus erythematosus nephritis, pulmonary emphysema, pulmonary ed ... The present invention relates to the treatment of various conditions, including pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, inflammatory lung disease, lymphocytic interstitial pneumonia, splenomegaly, type I diabetes, type II diabetes, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, hyperglycemia, arthritis, scleroderma, atopic dermatitis, lymphadenopathy, alopecia, hyperthyroidism, esophageal achalasia, thrombocytopenia, neutropenia, autoimmune hemolytic anemia, frontotemporal dementia, autism spectrum disorder, multiple sclerosis, or Friedreich's ataxia. In a preferred embodiment, the cancer is breast cancer. In another embodiment, the cancer is a glioma, such as glioblastoma multiforme.

[0016] According to a sixth aspect of the present invention there is provided a crystalline complex according to the first or second aspect of the present invention, or a pharmaceutical composition according to the fourth aspect of the present invention, for use in therapy in combination with one or more additional therapeutic agents. In one embodiment, the treatment is treatment of breast cancer and the additional therapeutic agent comprises an aromatase inhibitor, tamoxifen, exemestane, fulvestrant, an oral SERD or CDK4 / 6 inhibitor.

[0017] Detailed Description The disclosed composites, compositions, manufacturing processes, and methods may be more readily understood by reference to the following detailed description, which forms a part of this disclosure: It is to be understood that the disclosed composites, compositions, manufacturing processes, and methods are not limited to the specific composites, compositions, manufacturing processes, and methods described and / or illustrated herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only, and is not intended to be limiting of the claimed composites, compositions, manufacturing processes, and methods.

[0018] Reference to a particular value includes at least that particular value unless the context clearly dictates otherwise. When a range of values ​​is expressed, another embodiment includes from the one particular value and / or to the other particular value. Further, reference to values ​​stated in ranges includes each and every value within that range. All ranges are inclusive and combinable.

[0019] When values ​​are expressed as approximations, by use of the antecedent "about," it will be understood that the particular value forms another embodiment.

[0020] It will be understood that certain features of the disclosed composites, compositions, manufacturing processes, and methods, which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed composites, compositions, manufacturing processes, and methods, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0021] As used herein, the singular forms "a," "an," and "the" include the plural forms.

[0022] The term "about" used in reference to a numerical range, cutoff, or specific value indicates that the recited value may vary by up to about 10%. Many of the numerical values ​​used herein are determined experimentally, and those skilled in the art should understand that such determinations can and often do vary between different experiments. The values ​​used herein should not be considered unduly limited by this inherent variation. Thus, the term "about" is used to encompass variations of ±10% or less, ±5% or less, ±1% or less, ±0.5% or less, or ±0.1% or less from the stated value.

[0023] As used herein, "treating" and similar terms refers to reducing the severity and / or frequency of a symptom, eliminating said symptom and / or the underlying cause of said symptom, reducing the frequency or likelihood of the symptom and / or its underlying cause, delaying, preventing and / or slowing the progression of a disease and / or disorder, such as cancer or a benign proliferative disorder, and ameliorating or correcting the damage caused directly or indirectly by a disease and / or disorder, such as cancer or a benign proliferative disorder.

[0024] As used herein, the phrase "therapeutically effective amount" refers to an amount of a composition comprising at least one active pharmaceutical ingredient described herein effective to achieve a particular biological or therapeutic result, including, but not limited to, the biological or therapeutic results disclosed, described, or exemplified herein. A therapeutically effective amount may vary depending on factors such as the disease state, age, sex, and weight of the individual, and the ability of the composition to elicit a desired response in the subject. Such results include, but are not limited to, relief, remission, and / or regression of benign or malignant disease, or prevention of the onset of benign or malignant disease, as determined by any means appropriate in the art.

[0025] As used herein, a "subject" includes a vertebrate, a mammal, a domestic animal, or preferably a human.

[0026] Crystal complex Previous preparations of a 1:1 complex of sulforaphane and α-cyclodextrin have successfully isolated crystalline material, which typically exists as Form 1, containing 8-15% w / w water, or as a highly hydrated Form 2 containing approximately 13-20% w / w water, or as a mixture of these forms. Crystalline complexes prepared similarly to the method disclosed in WO 2013 / 179057 (Example 2) were found to be Form 1. The inventors have discovered a new crystalline form of sulforaphane and α-cyclodextrin complex (Form 3) that is less hygroscopic, has a lower water content, and, surprisingly, is more physically stable over a wider range of humidity conditions. Furthermore, while all crystalline forms isolated from sulforaphane and α-cyclodextrin complexes appear to be channel hydrates, Form 3 has surprisingly been found to have a more stable water content up to 60% RH at 25°C. Additional advantages that may be associated with the crystalline forms of the present invention include high form purity (e.g., having a single physical form) allowing for easier formulation, handling, and storage of the complex; good compaction for tablet formation; and rapid dissolution in biologically relevant media.

[0027] According to a first aspect of the present invention, there is provided a crystalline complex of sulforaphane and α-cyclodextrin having a water content of less than 8% w / w, wherein the crystalline form of the complex is stable at 25°C and between 0% and 60% relative humidity.

[0028] In one embodiment, the crystalline complex is a channel hydrate. Thus, a crystalline channel hydrate complex of sulforaphane and α-cyclodextrin is provided having a water content of less than 8% w / w, wherein the crystalline form of the complex is stable at 25°C and between 0% and 60% relative humidity.

[0029] The water content of a sample can be determined by any suitable means, as will be apparent to those skilled in the art. In one embodiment, the water content is determined by Karl Fischer titration. In one embodiment, the water content is determined by thermogravimetric analysis (TGA). In one embodiment, the water content is determined by gravimetric vapor sorption (GVS). After formation of a crystalline complex according to the present invention, the water content may be as low as 1-3% w / w. However, after exposing the complex to ambient conditions (25°C, 40% RH), the material typically equilibrates (e.g., over 3-7 days) to a water content of up to 5-7% w / w. However, Form 3, once equilibrated to 5-7% w / w water, has been found to maintain its physical form at 60% RH and to be unexpectedly stable in the 0%-60% humidity range. However, Form 1 exhibits an increased tendency to absorb additional water at 60% RH and convert to a mixture of forms (e.g., Form 1 and Form 2).

[0030] In one embodiment, the crystalline complex has a water content of less than 7.5% w / w, less than 7.0% w / w, less than 6.5% w / w, less than 6.0% w / w, less than 5.5% w / w, less than 5.0% w / w, or less than 4.5% w / w. In a preferred embodiment, the crystalline complex has a water content of less than 7% w / w. In a more preferred embodiment, the crystalline complex has a water content of less than 6% w / w.

[0031] In one embodiment, the crystalline complex comprises 1.5 to 7.5% w / w, 2.0 to 7.0% w / w, 2.5 to 7.5% w / w, 2.5 to 6.5% w / w, 2.5 to 6.0% w / w, 2.5 to 5.5% w / w, 3.0 to 8.0% w / w, 3.0 to 7.5% w / w, 3.0 to 6.5% w / w, 3.0 to 6.0% w / w, 3.0 to 5.5% w / w, 3.5 to 8.0% w / w, 3.5 to 7.5% w / w The crystalline composite has a water content of 1.0-7.9% w / w, such as 3.5-6.5% w / w, 3.5-6.0% w / w, 3.5-5.5% w / w, 4.0-8.0% w / w, 4.0-7.5% w / w, 4.0-6.5% w / w, 4.0-6.0% w / w, 4.0-5.5% w / w, 5.0-7.5% w / w, 5.0-7.0% w / w, 5.0-6.5% w / w, or 5.5-6.5% w / w. In one embodiment, the crystalline composite has a water content of 2.0-7.0% w / w.

[0032] It may be desirable to avoid over-drying the complex to avoid the potential for increased impurity formation. Thus, in a more preferred embodiment, the crystalline complex has a water content of 5.0-7.0% w / w, such as 5.5-6.5% w / w. In a preferred embodiment, the crystalline complex has a water content of about 6% w / w. In one embodiment, the water content, as determined by thermogravimetric analysis (TGA), is less than 6% w / w. In one embodiment, the water content, as determined by thermogravimetric analysis (TGA), is between 5.0-7.0% w / w, such as between 5.0-6.0% w / w.

[0033] As used herein, a crystalline form of a complex is "stable" if the crystalline form does not convert to another form under given conditions. In other words, stability relates to the physical stability of the solid form. This may be assessed by X-ray powder diffraction (XRPD) of a sample of the complex performed under given conditions. In one embodiment, the stability of the complex may be determined by variable humidity-XRPD, as described herein. In one embodiment, the crystalline form of the complex does not change between 25°C and 0% to 60% relative humidity when a sample of the complex is analyzed by variable humidity-XRPD. Thus, it will be appreciated that the crystalline form of the present invention has a broader stability domain compared to other forms of the complex. In one embodiment, the crystalline form of the complex is Form 3, and when a sample of the complex is analyzed by variable humidity-XRPD, the complex does not convert to Form 1 at 25°C and 0% to 60% relative humidity. In one embodiment, the crystalline form of the complex is Form 3, and when a sample of the complex is analyzed by variable humidity-XRPD, the complex does not convert to Form 2 at 25°C and 0% to 60% relative humidity.

[0034] In one embodiment, the physical form of a crystalline complex according to the present invention remains unchanged after storage at a temperature between 15°C and 25°C and a RH between 0% and 60%, as measured by XRPD, for at least one month. In a further embodiment, the physical form of a crystalline complex according to the present invention remains unchanged after storage at a temperature between 15°C and 25°C and a RH between 0% and 60%, as measured by XRPD, for at least two months (e.g., at least three months, at least four months, at least five months, or at least six months). In one embodiment, the physical form of a crystalline complex according to the present invention remains unchanged after storage at about 40°C and a RH between 0% and 60%, as measured by XRPD, for at least one month. In a further embodiment, the physical form of a crystalline complex according to the present invention remains unchanged after storage at about 40°C and a RH between 0% and 60%, as measured by XRPD, for at least two months (e.g., at least three months, at least four months, at least five months, or at least six months).

[0035] In another embodiment, the stability of a crystalline form according to the present invention may refer to the chemical stability of sulforaphane contained in the complex. Sulforaphane purity may be assessed against an internal standard by high-performance liquid chromatography (HPLC) of a sample of the complex dissolved in water, according to standard procedures known in the art. In one embodiment, the sulforaphane loading of a crystalline complex according to the present invention is greater than 90% after storage at 15-25°C and 0-60% RH for at least one month, where loading refers to the percentage of sulforaphane purity by HPLC relative to the purity measured at the beginning of the storage period (t=0). In a further embodiment, the sulforaphane loading of a crystalline complex according to the present invention is greater than 90% after storage at 15-25°C and 0-60% RH for at least two months (e.g., at least three months, at least four months, at least five months, or at least six months). In another embodiment, the sulforaphane loading of a crystalline complex according to the invention is greater than 90% after storage at 40°C and 0-60% RH for at least one month, where loading refers to the percentage of sulforaphane purity by HPLC relative to the purity measured at the beginning of the storage period (t=0). In a further embodiment, the sulforaphane loading of a crystalline complex according to the invention is greater than 90% after storage at 40°C and 0-60% RH for at least two months (e.g., at least three months, at least four months, at least five months, or at least six months). In another embodiment, the sulforaphane loading of a crystalline complex according to the invention is greater than 90% after storage in a sealed container at 40°C and 75% RH for at least one month, where loading refers to the percentage of sulforaphane purity by HPLC relative to the purity measured at the beginning of the storage period (t=0). In a further embodiment, the sulforaphane loading of a crystalline complex according to the invention is greater than 90% after storage in a sealed container at 40°C and 75% RH for at least 2 months (e.g., at least 3 months, at least 4 months, at least 5 months, or at least 6 months).

[0036] In one embodiment, a crystalline complex according to the first or second aspect of the invention comprises less than 5% total related impurities by HPLC after storage at 5° C. for at least 2 months (e.g., at least 3 months, at least 4 months, at least 5 months, or at least 6 months). Total related impurities refers to impurities related to sulforaphane and not other components of the complex, such as α-cyclodextrin, water, or solvent, as would be understood by one of skill in the art. In one embodiment, a crystalline complex according to the invention comprises less than 4% total related impurities by HPLC (e.g., less than 3%, less than 2%, or less than 1.5%) after storage at 5° C. for at least 2 months.

[0037] In one embodiment, the crystalline complex according to the first or second aspect of the invention comprises less than 5% total related impurities by HPLC after storage at 25° C. and 60% RH for at least 2 months (such as at least 3 months, at least 4 months, at least 5 months, or at least 6 months). In one embodiment, the crystalline complex according to the invention comprises less than 4% (such as less than 3%, less than 2.5%, less than 2%, or less than 1.5%) total related impurities by HPLC after storage at 25° C. and 60% RH for at least 2 months.

[0038] In one embodiment, the crystalline complex according to the first or second aspect of the invention comprises less than 0.25% by HPLC area of ​​a dimeric impurity having the following structure after storage at 5° C. for at least 2 months (such as at least 3 months, at least 4 months, at least 5 months, or at least 6 months): [ka]

[0039] In advantageous embodiments, the crystalline complex comprises less than 0.20% (such as less than 0.15%, less than 0.10%, or less than 0.05%) of the dimeric impurity by HPLC area after storage at 5° C. for at least 2 months. In one embodiment, the crystalline complex according to the invention comprises less than 0.40% (such as less than 0.30%, less than 0.25%, less than 0.20%, less than 0.15%, or less than 0.10%) of the dimeric impurity by HPLC area after storage at 25° C. and 60% RH for at least 2 months (such as at least 3 months, at least 4 months, at least 5 months, or at least 6 months). In advantageous embodiments, the crystalline complex comprises less than 0.35% (such as less than 0.30%, less than 0.25%, less than 0.20%, less than 0.15%, or less than 0.10%) of the dimeric impurity by HPLC area after storage at 25° C. and 60% RH for at least 2 months.

[0040] In one embodiment, the crystalline complex according to the first or second aspect of the invention comprises less than 1% α-cyclodextrin adduct by HPLC area after storage at 5° C. for at least 2 months (such as at least 3 months, at least 4 months, at least 5 months, or at least 6 months). In an advantageous embodiment, the crystalline complex comprises less than 0.8% α-cyclodextrin adduct by HPLC area (such as less than 0.7%, less than 0.6%, less than 0.5%, less than 0.4%, or less than 0.3%) after storage at 5° C. for at least 2 months. In one embodiment, the crystalline complex according to the invention comprises less than 2.0% α-cyclodextrin adduct by HPLC area after storage at 25° C. and 60% RH for at least 2 months (such as at least 3 months, at least 4 months, at least 5 months, or at least 6 months). In advantageous embodiments, the crystalline complex comprises less than 1.5% (such as less than 1.25%, less than 1.0%, less than 0.80%, less than 0.70%, or less than 0.50%) α-cyclodextrin adduct by HPLC area after storage at 25° C. and 60% RH for at least 2 months.

[0041] In one embodiment, a crystalline complex according to the first or second aspect of the present invention comprises less than 0.25% by HPLC area of ​​the dimeric impurity shown above and less than 1% by HPLC area of ​​an α-cyclodextrin adduct after storage at 5°C for at least 2 months (e.g., at least 3 months, at least 4 months, at least 5 months, or at least 6 months). In one embodiment, a crystalline complex according to the present invention comprises less than 0.40% by HPLC area of ​​a dimeric impurity having the above structure and less than 2.0% by HPLC area of ​​an α-cyclodextrin adduct after storage at 25°C and 60% RH for at least 2 months (e.g., at least 3 months, at least 4 months, at least 5 months, or at least 6 months). In a preferred embodiment, the molar ratio of sulforaphane to α-cyclodextrin in the complex is in the range of 0.8:1 to 1.2:1, such as in the range of 0.9:1 to 1.1:1. In a more preferred embodiment, the molar ratio of sulforaphane to α-cyclodextrin in the complex is about 1:1. Preferably, the molar ratio of sulforaphane to α-cyclodextrin in the complex is 1: 1. In alternative embodiments, the molar ratio of sulforaphane to α-cyclodextrin in the complex is in the range of 0.2: 1 to 0.9: 1, such as in the range of 0.4: 1 to 0.6: 1, or about 0.5: 1.

[0042] In a preferred embodiment, the sulforaphane present in the complex of the first aspect is racemic sulforaphane. Accordingly, a crystalline complex of racemic sulforaphane and α-cyclodextrin having a water content of less than 8% w / w is provided, wherein the crystalline form of the complex is stable at 25°C and between 0% and 60% relative humidity. In one embodiment, a crystalline complex of racemic sulforaphane and α-cyclodextrin having a water content of less than 8% w / w is provided, wherein the molar ratio of sulforaphane to α-cyclodextrin in the complex is in the range of 0.9:1 to 1.1:1, and the crystalline form of the complex is stable at 25°C and between 0% and 60% relative humidity. In one embodiment, a crystalline complex of racemic sulforaphane and α-cyclodextrin is provided having a water content of less than 6% w / w, wherein the molar ratio of sulforaphane to α-cyclodextrin in the complex is 1:1, and the crystalline form of the complex is stable between 0% and 60% relative humidity at 25°C.

[0043] According to a second aspect of the present invention, there is provided a crystalline complex of sulforaphane and α-cyclodextrin (Form 3), wherein the crystalline form of the complex is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ, when measured in reflectance mode.

[0044] As used herein, reference to Form 3 of the sulforaphane and α-cyclodextrin complex refers to a complex having a molar ratio of sulforaphane to α-cyclodextrin of about 1:1. Determining the molar ratio is: 1 This is performed according to H-NMR spectroscopy. Due to the inherent error limit in calculating the ratio, the molar ratio of sulforaphane to α-cyclodextrin in the complex may range from 0.9:1 to 1.1:1.

[0045] The crystalline form of the complex may be analyzed by XRPD in reflectance or transmission mode. The crystalline form of the complex according to the second aspect is characterized by peaks at 5.3 and 10.7±0.2 degrees 2θ when measured by XRPD in reflectance mode. In one embodiment, the crystalline complex according to the second aspect is further characterized by additional XRPD peaks at 8.1 and 16.1±0.2 degrees 2θ when measured in reflectance mode.

[0046] In one embodiment, the crystalline complex of sulforaphane and α-cyclodextrin (Form 3) exhibits an X-ray powder diffraction pattern, measured at room temperature using Cu Kα radiation, substantially the same as the X-ray powder diffraction pattern shown in Figure 4. In one embodiment, the crystalline complex of sulforaphane and α-cyclodextrin (Form 3) exhibits an X-ray powder diffraction pattern in reflection mode, measured at room temperature using Cu Kα radiation, substantially the same as the X-ray powder diffraction pattern shown in Figure 4A. In one embodiment, the crystalline complex of sulforaphane and α-cyclodextrin (Form 3) exhibits an X-ray powder diffraction pattern in transmission mode, measured at room temperature using Cu Kα radiation, substantially the same as the X-ray powder diffraction pattern shown in Figure 4B.

[0047] The term "substantially the same" in reference to XRPD means that variations in peak positions and relative intensities of the peaks are taken into account. For example, for 2θ values, typical accuracy is within ±0.2° 2θ. One skilled in the art will understand that relative peak intensities will exhibit instrument-to-instrument variations as well as variations due to crystallinity, preferred orientation, sample preparation, and other factors known in the art.

[0048] In a preferred embodiment, a Form 3 crystalline complex of racemic sulforaphane and α-cyclodextrin is provided, wherein the crystalline form of the complex is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflectance mode. Most preferably, a crystalline complex of racemic sulforaphane and α-cyclodextrin is provided, wherein the complex has a molar ratio of sulforaphane to α-cyclodextrin of about 1:1 and is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflectance mode. Most preferably, a crystalline complex of racemic sulforaphane and α-cyclodextrin is provided, wherein the complex has a molar ratio of sulforaphane to α-cyclodextrin of about 1:1; a water content of less than 8% w / w (such as less than 6% w / w, or between 5-7% w / w); and is characterized by XRPD peaks at 5.3 and 10.7±0.2°2θ when measured in reflector mode.

[0049] The crystalline complex of the present invention may advantageously be present in particulate form. When formed by the preferred process described herein comprising agitation drying, the crystalline complex according to the first or second aspect of the present invention is formed with a low level of agglomerated particles (typically referring to particles greater than 2000 μm (2 mm) in diameter). The degree of agglomeration may be determined by passing a sample of the material through a 2000 micron sieve and quantifying the excess residue relative to the input amount (% w / w).

[0050] In one embodiment, a crystalline complex according to the invention is provided, wherein the complex comprises less than 5% w / w of particles greater than 2000 μm in diameter. In an embodiment, a crystalline complex according to the invention is provided, wherein the complex comprises less than 4.5% w / w (such as less than 4.0% w / w, less than 3.5% w / w, or less than 3.0% w / w) of particles greater than 2000 μm in diameter.

[0051] manufacturing process In a third aspect, there is provided a method for forming a crystalline complex of sulforaphane and α-cyclodextrin according to the first or second aspect, the method comprising: a) providing a crystalline complex of sulforaphane and α-cyclodextrin; b) drying the composite from step a) while stirring the composite at a pressure of less than 200 mbar until the moisture content of the composite is less than 6% w / w; The compound comprises:

[0052] Sulforaphane can be derived from natural sources or prepared by synthetic procedures. Sulforaphane derived from natural sources is chiral and exists predominantly in the (R) configuration. In step a), the sulforaphane is preferably racemic sulforaphane. Racemic sulforaphane can be synthesized by various known methods, such as those disclosed by Schmid and Karrer (Helvetica Chimica Acta (1948), 1497), WO 2008 / 091608, or WO 2013 / 179057.

[0053] A complex of racemic sulforaphane and α-cyclodextrin can be prepared according to Example 1 or as disclosed in WO 2013 / 179057. Typically, the molar ratio of sulforaphane to α-cyclodextrin in the complex is about 1:1. In a preferred embodiment, step a) comprises providing a crystalline complex of racemic sulforaphane and α-cyclodextrin, wherein the complex has a molar ratio of sulforaphane to α-cyclodextrin of 1:1.

[0054] In one embodiment, the crystalline complex of sulforaphane and α-cyclodextrin provided in step a) exists as Form 1. Form 1 is characterized by a peak at 9.8±0.2°2θ as measured by XRPD in reflectance mode. Typically, Form 1 contains 8-15% w / w water. In one embodiment, the crystalline complex of sulforaphane and α-cyclodextrin provided in step a) contains more than 8% w / w water, such as more than 10% w / w water or more than 12% w / w water. In one embodiment, the crystalline complex of sulforaphane and α-cyclodextrin provided in step a) contains 10-15% w / w water, such as 12-14% w / w water. In one embodiment, the crystalline complex of sulforaphane and α-cyclodextrin provided in step a) exists as Form 2. Form 2 is characterized by peaks at 9.5, 14.2, and 23.6±0.2°2θ as measured by XRPD in reflectance mode. Typically, Form 2 contains 13-20% w / w water. In one embodiment, the crystalline complex of sulforaphane and α-cyclodextrin provided in step a) exists as a mixture of Form 1 and Form 2.

[0055] It has been found that using a very wet crystalline complex as input to the drying process can result in balling or agglomeration problems in step b) of the process. Therefore, advantageously, the crystalline complex of sulforaphane and α-cyclodextrin provided in step a) has a water content of 25% w / w or less, such as less than 25% w / w, less than 24% w / w, less than 23% w / w, less than 22% w / w, less than 21% w / w, or less than 20% w / w. In one embodiment, the complex provided in step a) has a water content of 8-25% w / w, such as 10-25% w / w, 10-22% w / w, or 10-20% w / w.

[0056] Thus, if the material provided in step a) has a water content of more than 25% w / w, preferably the complex is dried without agitation over an initial period prior to step b) until the complex has a water content of 25% w / w or less, such as 20% w / w or less.

[0057] The drying in step b) is carried out at a pressure of less than 200 mbar. In an advantageous embodiment, the drying in step b) is carried out at a pressure of less than 150 mbar, such as less than 100 mbar, less than 80 mbar or less than 75 mbar. In a most advantageous embodiment, the drying in step b) is carried out at a pressure of about 50-70 mbar.

[0058] In one embodiment, the drying in step b) is carried out under a flow of inert gas. In a convenient embodiment, the inert gas is nitrogen. Conveniently, the inert gas (such as nitrogen) is supplied at a flow rate of 0.1 to 1.0 L / min, such as 0.3 to 0.6 L / min, or 0.4 to 0.5 L / min.

[0059] The drying of step b) is carried out while stirring the complex. The stirring may be carried out by any suitable means. Static (unstirred) drying typically results in inefficient formation of the crystalline form of the invention, especially at large scales where sample heterogeneity and particle aggregation may be problematic. In a preferred embodiment, the drying of step b) is carried out with constant or substantially constant stirring. By "substantially constant" it is meant that the stirring may be stopped for short periods of time so that material can be removed, for example to check the moisture content. In one embodiment, the drying of step b) is carried out with stirring for more than 80%, such as more than 90% or more than 95% of the duration of the drying step.

[0060] In one embodiment, the stirring in step b) is provided at a speed of more than 5 rpm. In a more advantageous embodiment, the stirring is provided at a speed of more than 10 rpm, such as 20-40 rpm. In an advantageous embodiment, the drying in step b) is carried out using overhead stirring. The overhead stirring may be used to agitate the filter cake in a suitable drying apparatus, such as a Nutsche filter dryer. In an advantageous embodiment, the overhead stirring is provided at a speed of more than 5 rpm. In a more advantageous embodiment, the overhead stirring is provided at a speed of more than 10 rpm, such as 20-40 rpm.

[0061] In a preferred embodiment, the drying in step b) is carried out for at least 5 hours, such as at least 6 hours or at least 7 hours. In one embodiment, the drying in step b) is carried out for 5 to 24 hours, such as for 5 to 12 hours or for 7 to 12 hours. In one embodiment, the drying in step b) is carried out for about 9 to 10 hours.

[0062] Because sulforaphane can decompose at elevated temperatures, drying is advantageously carried out at ambient temperature. In a preferred embodiment, drying in step b) is carried out at a temperature between 10° C. and 30° C. In one embodiment, drying in step b) is carried out at a temperature between 10° C. and 25° C., or between 15° C. and 30° C., such as between 15° C. and 25° C., or about 20° C.

[0063] In one embodiment, the drying in step b) is carried out at a temperature between 10°C and 30°C for 5 to 12 hours. In one embodiment, the drying in step b) is carried out at a temperature between 10°C and 30°C for 5 to 12 hours with substantially constant agitation. In one embodiment, the drying in step b) is carried out under a flow of inert gas at a temperature between 10°C and 30°C for 5 to 12 hours with substantially constant agitation. In one embodiment, the drying in step b) is carried out under a flow of inert gas and a pressure of less than 100 mbar at a temperature between 10°C and 30°C for 5 to 12 hours with substantially constant agitation.

[0064] The crystalline complex of sulforaphane and α-cyclodextrin according to the present invention contains less than 8% w / w water, such as 5-7% w / w water. To ensure conversion to the complex according to the present invention, it is necessary to dry the complex from step a) until the water content of the complex is less than 6% w / w. In one embodiment, drying in step b) is carried out until the water content of the complex is less than 5% w / w, such as less than 4% w / w, less than 3% w / w, or less than 2% w / w. In a preferred embodiment, drying in step b) is carried out until the water content of the complex is between 4% and 6% w / w, such as between 5% and 5.5% w / w.

[0065] Form 3 is characterized by a major XRPD peak at 10.7±0.2°2θ when measured in reflectance mode. If the sample is overdried (e.g., less than approximately 4% w / w water), the major XRPD peak may be greater than 10.9°2θ when measured in reflectance mode. However, after drying is complete, exposure of the complex to ambient conditions will result in a slow equilibration of the water content to approximately 5-7% w / w, with the major XRPD peak coinciding with 10.7±0.2°2θ.

[0066] The drying process described above may be carried out more efficiently if the surface area of ​​the composite is maximized. Thus, optionally prior to step b), the surface area of ​​the crystalline composite may be increased by de-clumping and / or other comminution processing of the material. In one embodiment, prior to step b), the composite is subjected to a further processing step (such as milling or grinding) that increases the surface area of ​​the composite.

[0067] As can be seen from the large-scale formation of Form 3 complexes described in Example 5, without agitation, conversion to Form 3 does not occur unless Form 1 is dried for extended periods of time, optionally at elevated temperatures and under a sufficiently strong vacuum. The filter drying studies described in Example 5B demonstrate that Form 3 is reproducibly obtained after drying at ambient temperature under vacuum for less than 12 hours when the drying step is carried out with essentially constant agitation of the Form 1 complexes.

[0068] In one embodiment, there is provided a crystalline complex of sulforaphane and α-cyclodextrin obtainable or obtained by the process described herein.

[0069] In certain embodiments, a) providing a crystalline complex of sulforaphane and α-cyclodextrin; b) drying the complex from step a) under a flow of inert gas at a pressure of less than 100 mbar and at a temperature between 10°C and 30°C with substantially constant agitation until the water content of the complex is less than 6% w / w; The present invention provides a crystalline complex of sulforaphane and α-cyclodextrin that is obtainable or obtained by a process comprising:

[0070] composition According to a fourth aspect of the present invention, there is provided a solid pharmaceutical composition comprising an effective amount of a crystalline complex according to the first or second aspect of the present invention, and optionally at least one pharmaceutically acceptable excipient.

[0071] In one embodiment, the pharmaceutical composition comprises a crystalline complex of sulforaphane and α-cyclodextrin, wherein greater than 50% of the complex is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflectance mode. Preferably, greater than 55%, 60%, 65%, 75%, 80%, or 85% of the complex is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflectance mode. In a most preferred embodiment, greater than 90% of the complex is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflectance mode. The percentage of the complex present in a particular form may be approximated by XRPD or Raman spectroscopy following construction of a calibration curve.

[0072] Over-drying Form 3 may result in a complex having the crystalline structure of Compacted Form 3 (e.g., a decrease in the distance between crystal lattice layers). Such Compacted Form 3 may have a major XRPD peak at 10.9±0.2 degrees 2θ when measured in reflectance mode. Because Compacted Form 3 tends to gradually equilibrate back to Form 3 unless kept under anhydrous conditions, mixtures of Form 3 and Compacted Form 3 have the potential to provide the beneficial properties of Form 3 described herein. In one embodiment, a pharmaceutical composition comprises a crystalline complex of sulforaphane and α-cyclodextrin, wherein the complex comprises a mixture of Form 3 and Compacted Form 3. In one embodiment, a pharmaceutical composition comprises a crystalline complex of sulforaphane and α-cyclodextrin, wherein the complex is characterized by XRPD peaks at 10.7 and 10.9±0.2 degrees 2θ when measured in reflectance mode. In one embodiment, greater than 60% of the complex is Form 3 (major XRPD peak at 10.7±0.2 degrees 2θ) and less than 40% is compressed Form 3 (major XRPD peak at 10.9±0.2 degrees 2θ). Suitably, greater than 80% of the complex is Form 3 (major XRPD peak at 10.7±0.2 degrees 2θ) and less than 20% is compressed Form 3 (major XRPD peak at 10.9±0.2 degrees 2θ).

[0073] Pharmaceutical compositions according to the present invention may be administered by any suitable means, which can be determined by one skilled in the art based on the disease or disorder being treated with the composition.

[0074] The solid compositions of the present invention may be in a form suitable for oral use (e.g., tablets, lozenges, hard or soft capsules, dispersible powders or granules), administration by inhalation (e.g., finely divided powder), or administration by insufflation (e.g., finely divided powder). Preferably, the solid pharmaceutical composition is for oral administration.

[0075] The compositions of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more coloring agents, sweeteners, flavoring agents and / or preservatives.

[0076] As used herein, "pharmaceutically acceptable excipient" means a pharmaceutically acceptable substance, composition, or vehicle that is involved in giving form or consistency to a pharmaceutical composition. Each excipient must be compatible with the other ingredients of the pharmaceutical composition when mixed, so as to avoid interactions that would substantially reduce the efficacy of the conjugate of the present invention when administered to a patient, and interactions that would result in a pharmaceutically unacceptable pharmaceutical composition. In addition, of course, each excipient must be of sufficiently high purity so as to be pharmaceutically acceptable.

[0077] Suitable pharmaceutically acceptable excipients vary depending on the particular dosage form selected. Furthermore, suitable pharmaceutically acceptable excipients may be selected for the particular function they may serve in the composition. For example, certain pharmaceutically acceptable excipients may be selected for their ability to facilitate the production of a uniform dosage form. Certain pharmaceutically acceptable excipients may be selected for their ability to facilitate the production of a stable dosage form. Certain pharmaceutically acceptable excipients may be selected for their ability to facilitate the transport or transfer of the complexes or sulforaphane of the present invention from one organ or part of the body to another organ or part of the body once administered to a patient. Certain pharmaceutically acceptable excipients may be selected for their ability to improve patient compliance.

[0078] Suitable pharmaceutically acceptable excipients include the following excipient types: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavoring agents, flavor-masking agents, colorants, anti-caking agents, humectants, chelating agents, plasticizers, thickeners, antioxidants, preservatives, stabilizers, surfactants, and buffers. One of ordinary skill in the art will understand that a particular pharmaceutically acceptable excipient may serve more than one function, or may serve alternative functions, depending on the amount of the excipient present in the formulation and the other ingredients present in the formulation.

[0079] Those skilled in the art possess the knowledge and skill in the art to be able to select appropriate pharmaceutically acceptable excipients in appropriate amounts for use in the present invention. Furthermore, there are many resources available to those skilled in the art that describe pharmaceutically acceptable excipients and may be useful in selecting appropriate pharmaceutically acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

[0080] The pharmaceutical compositions of the present invention are prepared using techniques and methods known to those skilled in the art, some of which are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

[0081] An effective amount of the conjugate of the invention for use in treating a disease or disorder is an amount sufficient to symptomatically alleviate the symptoms of the disease or disorder, slow the progression of the disease or disorder, or reduce the risk of deterioration in a patient with symptoms of the disease or disorder in a warm-blooded animal, particularly a human.

[0082] The amount of active conjugate that is combined with one or more excipients to produce a single dosage form will necessarily vary depending upon the host treated and the particular route of administration. For example, a formulation intended for oral administration to humans will generally contain from 0.5 mg to 1.0 g (more preferably 100 to 500 mg, e.g., 300 mg) of active conjugate, compounded with an appropriate and convenient amount of excipient, which may vary, for example, from about 5 to about 98% by weight of the total composition.

[0083] The magnitude of an administered dose of a conjugate of the invention for therapeutic or prophylactic purposes will necessarily vary according to well-known principles of medicine, depending on the nature and severity of the condition, the age, weight, sex, and diet of the subject, and the route of administration.

[0084] When the complex of the present invention is used for therapeutic or prophylactic purposes, it is generally administered in a daily dose ranging, for example, from 0.1 mg to 75 mg per kg of body weight, given in divided doses as needed. Oral administration may be suitable, particularly in tablet form. Typically, a unit dosage form contains about 0.5 mg to 1.0 g of the complex of the present invention. In a preferred embodiment, the unit dosage form of the composition contains about 100 mg to 500 mg, such as about 200 mg to 400 mg, of the complex of the present invention. In a most preferred embodiment, the unit dosage form of the composition contains about 300 mg of the complex of the present invention. The unit dosage form for oral administration may be a tablet or a capsule. In a preferred embodiment, the solid pharmaceutical composition of the present invention is formulated as a tablet. In an alternative preferred embodiment, the solid pharmaceutical composition of the present invention is formulated as a capsule.

[0085] The daily dose may be administered as a single dose. Alternatively, administration may be two or more times daily. By way of example, the pharmaceutical composition (e.g., as a tablet or capsule) may be orally administered at least once daily, such as once daily, or twice daily.

[0086] Suitable amounts of the active complex to be administered as a daily dose are about 1 mg to about 1 g, such as about 5 mg to about 2 g, about 10 mg to about 1 g, such as about 5 mg to about 500 mg, such as about 10 mg to about 500 mg, such as about 10 mg to about 400 mg, such as about 200 mg to about 400 mg, such as about 250 mg to about 350 mg, such as about 280 mg to about 320 mg, such as about 290 mg to about 310 mg, such as about 300 mg, such as 300 mg, such as about 50 mg to about 900 mg, such as about 100 mg to about 800 mg, such as about 300 mg to about 700 mg, such as about 500 mg to about 700 mg, such as about 600 mg, or 600 mg, such as about 1 mg to about 5 g.

[0087] medical use According to a fifth aspect of the present invention there is provided a crystalline complex according to the first or second aspect of the present invention, or a pharmaceutical composition according to the fourth aspect of the present invention, for use in therapy.

[0088] In one embodiment there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use as a medicament.

[0089] In one embodiment, the pharmaceutical composition is for use as a medicament, wherein the pharmaceutical composition is administered orally.

[0090] In one embodiment, the complex or pharmaceutical composition is useful for the treatment and / or prevention of a disease and / or disorder. In one embodiment, there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use in treating a disease or disorder mediated by Nrf2 or STAT3. In one embodiment, there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use in treating a disease or disorder mediated by Nrf2 activation or STAT3 inactivation.

[0091] In another embodiment, the present invention relates to the use of a complex or composition of the invention as defined herein in the manufacture of a medicament for use in the treatment of a disease or disorder mediated by Nrf2 activation or STAT3 inactivation.

[0092] In another embodiment, the present invention relates to a method for treating a disease or disorder mediated by Nrf2 activation or STAT3 inactivation, said method comprising administering a therapeutically effective amount of a complex or composition of the present invention as defined herein to a subject in need of such treatment.

[0093] In one embodiment, the therapeutic target for the treatment of cancer, subarachnoid hemorrhage, intracerebral hemorrhage, ischemic stroke, delayed cerebral ischemia, atherosclerosis, middle cerebral artery infarction, pulmonary arterial hypertension, Alport syndrome, non-alcoholic steatohepatitis, focal segmental glomerulosclerosis, Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, cirrhosis, non-alcoholic fatty liver disease, hepatitis, acute kidney injury, sepsis, chronic kidney disease, systemic lupus erythematosus nephritis, emphysema, pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, inflammatory lung disease, lymphocytic interstitial pneumonia, pulmonary fibrosis, pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, inflammatory lung disease, lymphocytic fibrosis, pulmonary ... In accordance with the present invention, there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use in the treatment of interstitial pneumonia, splenomegaly, type I diabetes, type II diabetes, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, hyperglycemia, arthritis, scleroderma, atopic dermatitis, lymphadenopathy, alopecia, hyperthyroidism, oesophageal achalasia, thrombocytopenia, neutropenia, autoimmune hemolytic anemia, frontotemporal dementia, autism spectrum disorder, multiple sclerosis or Friedreich's ataxia.

[0094] In one embodiment there is provided a complex or composition of the invention as defined herein for use in the treatment of cancer, subarachnoid hemorrhage, intracerebral hemorrhage, ischemic stroke, delayed cerebral ischemia, pulmonary arterial hypertension, Alport syndrome, non-alcoholic steatohepatitis, focal segmental glomerulosclerosis, Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, frontotemporal dementia, autism spectrum disorder, multiple sclerosis or Friedreich's ataxia.

[0095] In another embodiment, the present invention provides a method for treating cancer, subarachnoid hemorrhage, intracerebral hemorrhage, ischemic stroke, delayed cerebral ischemia, atherosclerosis, middle cerebral artery infarction, pulmonary arterial hypertension, Alport syndrome, non-alcoholic steatohepatitis, focal segmental glomerulosclerosis, Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, cirrhosis, non-alcoholic fatty liver disease, hepatitis, acute kidney injury, sepsis, chronic kidney disease, systemic lupus erythematosus nephritis, emphysema, pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, and inflammatory lung diseases. , lymphocytic interstitial pneumonia, splenomegaly, type I diabetes, type II diabetes, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, hyperglycemia, arthritis, scleroderma, atopic dermatitis, lymphadenopathy, alopecia, hyperthyroidism, oesophageal achalasia, thrombocytopenia, neutropenia, autoimmune hemolytic anemia, frontotemporal dementia, autism spectrum disorder, multiple sclerosis or Friedreich's ataxia.

[0096] In another embodiment, the present invention provides a method for treating cancer, subarachnoid hemorrhage, intracerebral hemorrhage, ischemic stroke, delayed cerebral ischemia, atherosclerosis, middle cerebral artery infarction, pulmonary arterial hypertension, Alport syndrome, nonalcoholic steatohepatitis, focal segmental glomerulosclerosis, Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, cirrhosis, nonalcoholic fatty liver disease, hepatitis, acute kidney injury, sepsis, chronic kidney disease, systemic lupus erythematosus nephritis, emphysema, pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, inflammatory lung disease, lymphocytic interstitial pneumonia. , splenomegaly, type I diabetes, type II diabetes, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, hyperglycemia, arthritis, scleroderma, atopic dermatitis, lymphadenopathy, alopecia, hyperthyroidism, esophageal achalasia, thrombocytopenia, neutropenia, autoimmune hemolytic anemia, frontotemporal dementia, autism spectrum disorder, multiple sclerosis or Friedreich's ataxia, the method comprising administering to a subject in need of such treatment a therapeutically effective amount of a complex or composition of the invention as defined herein.

[0097] In one embodiment there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use in the treatment of cancer, subarachnoid hemorrhage, delayed cerebral ischemia, non-alcoholic steatohepatitis, or autism spectrum disorder.

[0098] In another embodiment, the invention relates to the use of a conjugate or composition of the invention as defined herein in the manufacture of a medicament for use in the treatment of cancer, subarachnoid hemorrhage, delayed cerebral ischemia, non-alcoholic steatohepatitis, or autism spectrum disorder.

[0099] In another embodiment, the present invention relates to a method for treating cancer, subarachnoid hemorrhage, delayed cerebral ischemia, non-alcoholic steatohepatitis, or autism spectrum disorder, the method comprising administering to a subject in need of such treatment a therapeutically effective amount of a complex or composition of the present invention as defined herein.

[0100] The cancer to be treated may be a solid tumor (such as breast cancer, colorectal cancer, lung cancer, liver cancer, bladder cancer, cervical cancer, hepatocellular carcinoma, squamous cell carcinoma, melanoma, glioma, head and neck cancer, pancreatic cancer, or prostate cancer) or a hematological cancer (such as leukemia, acute lymphoblastic leukemia, juvenile myelomonocytic leukemia, non-Hodgkin's lymphoma, or diffuse large B-cell lymphoma). In a preferred embodiment, the cancer is breast cancer. In a most preferred embodiment, the breast cancer is ER+ or HER2- metastatic breast cancer. In another embodiment, the cancer is a glioma, such as high-grade glioma or glioblastoma multiforme.

[0101] combination The conjugates and compositions of the invention may be used as monotherapy or in combination with other compounds or treatments to prevent or treat a disease or disorder.

[0102] Thus, according to a sixth aspect of the invention there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use in therapy in combination with one or more additional therapeutic agents, the choice of which will, of course, depend on the disease or condition to be treated and its severity.

[0103] The use of combination therapies to treat certain medical conditions is common.

[0104] As used herein, when the term "combination" is used, it should be understood that this refers to simultaneous, separate or sequential administration. In one aspect of the present invention, "combination" refers to simultaneous administration. In another aspect of the present invention, "combination" refers to separate administration. In a further aspect of the present invention, "combination" refers to sequential administration. When administration is sequential or separate, the delay in administration of the second component should not be such that the beneficial effect of the combination is lost.

[0105] In one embodiment, there is provided a combination comprising the complex or composition of the invention as defined above and another therapeutic agent, suitable for use in the treatment of a disease or condition involving activation of Nrf2 or inactivation of STAT3.

[0106] In one embodiment there is provided a combination suitable for use in the prevention or treatment of cancer, subarachnoid hemorrhage, intracerebral hemorrhage, ischemic stroke, delayed cerebral ischemia, pulmonary arterial hypertension, Alport syndrome, non-alcoholic steatohepatitis, focal segmental glomerulosclerosis, Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, frontotemporal dementia, autism spectrum disorder, multiple sclerosis or Friedreich's ataxia, the combination comprising a complex or composition of the invention as defined above and one or more additional therapeutic agents.

[0107] In one embodiment there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use in the treatment of cancer in combination with one or more additional therapeutic agents and / or in combination with one or more additional treatments (e.g. radiation therapy).

[0108] In one embodiment there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use in the treatment of breast cancer in combination with one or more additional therapeutic agents selected from an aromatase inhibitor, tamoxifen, exemestane, fulvestrant, an oral SERD or CDK4 / 6 inhibitor.

[0109] In one embodiment there is provided a combination suitable for use in the prevention or treatment of breast cancer, the combination comprising a complex or composition of the invention as defined above and one or more additional therapeutic agents selected from an aromatase inhibitor, tamoxifen, exemestane, fulvestrant, an oral SERD or CDK4 / 6 inhibitor.

[0110] In one embodiment there is provided a crystalline complex according to the first or second aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention, for use in the treatment of glioma (such as glioblastoma multiforme) in combination with one or more of radiation therapy and temozolomide.

[0111] In one embodiment there is provided a combination suitable for use in the prevention or treatment of glioma (such as glioblastoma multiforme), the combination comprising a complex or composition of the invention as defined above and one or more of radiation therapy and temozolomide.

[0112] The following numbered statements refer to certain aspects and embodiments of the present disclosure, although they are not claims. 1. A crystalline complex of sulforaphane and α-cyclodextrin having a water content of less than 8% w / w, wherein the crystalline form of the complex is stable at 25°C and between 0% and 60% relative humidity. 10. The crystalline complex of statement 1 having a water content of less than 2.6% w / w. 3. The crystalline complex of claim 1 or 2, wherein the molar ratio of sulforaphane to α-cyclodextrin in the complex is in the range of 0.9:1 to 1.1:1. 4. The crystalline complex of statement 1 or statement 2, wherein the molar ratio of sulforaphane to α-cyclodextrin in the complex is about 1:1. 5. Crystalline complex of sulforaphane with α-cyclodextrin (Form 3), wherein the crystalline form of the complex is characterized by XRPD peaks at 5.3 and 10.7±0.2°2θ when measured in reflectance mode. 6. The crystalline complex of statement 5, further characterized by additional XRPD peaks at 8.1 and 16.1±0.2 degrees 2θ when measured in reflectance mode. 7. A crystalline complex according to statement 5, which exhibits an X-ray powder diffraction pattern substantially the same as the X-ray powder diffraction pattern shown in Figure 4 when measured at room temperature using Cu Kα radiation. 8.a) providing a complex of sulforaphane and α-cyclodextrin; b) drying the composite from step a) at a temperature of not less than 25°C and a pressure of less than 50 mbar for at least 12 hours until the moisture content of the composite is less than 6% w / w; 8. A method for forming a crystalline complex of sulforaphane and α-cyclodextrin as described in any one of statements 1 to 7, comprising: 9. The method of statement 8, wherein the drying of step b) is carried out at a pressure of less than 10 mbar. 10. The method of claim 8 or claim 9, wherein the drying of step b) is carried out for at least 18 hours. 11. The method of any one of claims 8 to 10, wherein the drying in step b) is carried out at a temperature between 25°C and 30°C for 24 to 200 hours. 12. The method of any one of claims 8 to 11, wherein prior to step b), the composite is subjected to a further processing step that increases the surface area of ​​the composite. 13. A crystalline complex of sulforaphane and α-cyclodextrin obtained by the method according to any one of claims 8 to 12. 14. A solid pharmaceutical composition comprising an effective amount of the crystalline complex of any one of statements 1-7 or 13, and optionally at least one pharmaceutically acceptable excipient. 15. The solid pharmaceutical composition of statement 14, wherein greater than 50% of the crystalline complex of sulforaphane and α-cyclodextrin is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflectance mode. 16. The solid pharmaceutical composition of statement 14, wherein greater than 80%, such as greater than 90%, of the crystalline complex of sulforaphane and α-cyclodextrin is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflectance mode. 17. The solid pharmaceutical composition of statements 14-16, wherein the composition is formulated as a tablet. 18. The solid pharmaceutical composition of statements 14-16, wherein the composition is formulated as a capsule. 19. A conjugate according to statements 1 to 7 or 13, or a pharmaceutical composition according to statements 14 to 18, for use in therapy. 20. A conjugate according to statements 1 to 7 or 13, or a pharmaceutical composition according to statements 14 to 18, for use in treating a disease or disorder mediated by Nrf2 or STAT3. 21. Cancer, subarachnoid hemorrhage, intracerebral hemorrhage, ischemic stroke, delayed cerebral ischemia, atherosclerosis, middle cerebral artery infarction, pulmonary arterial hypertension, Alport syndrome, nonalcoholic steatohepatitis, focal segmental glomerulosclerosis, Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, cirrhosis, nonalcoholic fatty liver disease, hepatitis, acute kidney injury, sepsis, chronic kidney disease, systemic lupus erythematosus nephritis, emphysema, pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, inflammatory lung disease, phosphorus 19. A conjugate according to statements 1 to 7 or 13, or a pharmaceutical composition according to statements 14 to 18, for use in the treatment of leukemia, pulmonary fibrosis, splenomegaly, type 1 diabetes, type 2 diabetes, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, hyperglycemia, arthritis, scleroderma, atopic dermatitis, lymphadenopathy, alopecia, hyperthyroidism, esophageal achalasia, thrombocytopenia, neutropenia, autoimmune hemolytic anemia, frontotemporal dementia, autism spectrum disorder, multiple sclerosis or Friedreich's ataxia. 22. The use according to statement 21, wherein the cancer is breast cancer. 23. The use of statement 22, wherein the breast cancer is ER+ or HER2- metastatic breast cancer. 24. A conjugate according to statements 1-7 or 13, or a pharmaceutical composition according to claims 14-18, in combination with one or more additional therapeutic agents. 25. The combination of statement 24, wherein the additional therapeutic agent comprises an aromatase inhibitor, tamoxifen, exemestane, fulvestrant, an oral SERD or CDK4 / 6 inhibitor.

[0113] Example Specific embodiments of the present invention are further described below with reference to the accompanying drawings. [Brief explanation of the drawings]

[0114] [Figure 1] 1 is an XRPD analysis in reflector mode of Example 1 (Form 1) performed on a Panalytical X'pert instrument. [Figure 2] Figure 1 shows an XRPD overlay in both reflection and transmission mode of Form 1 performed on a Panalytical Empyrean instrument. [Figure 3] Figure 1 shows an XRPD overlay in both reflection and transmission mode of Example 2 (Form 2) performed on a Panalytical Empyrean instrument. [Figure 4] Figure 1 shows XRPD overlays in both reflection (A) and transmission (B) modes of Example 3 (Form 3) performed on a Panalytical Empyrean instrument. [Figure 5] 1 shows XRPD overlays performed in reflectance mode for Configurations 1, 2, and 3 performed on a Panalytical Empyrean instrument (Configurations 2 and 3) and a Bruker AXSD8 instrument (Configuration 1). [Figure 6] 1 shows XRPD overlays in reflectance mode of Form 1 and Example 3 (Form 3) after Example 3 was exposed to ambient conditions for 0, 7, 9, and 29 days, performed on a Panalytical Empyrean instrument. [Figure 7] 1 shows the H-NMR spectrum of Example 3 carried out at 400 MHz in DMSO-d. [Figure 8] (A) XRPD analysis of Example 4 in transmission mode performed on a Panalytical X'pert Pro instrument, and (B) XRPD overlay of Forms 1 and 3 (in both reflection and transmission modes) with Example 4 (in transmission mode). [Figure 9] Figure 1 shows a variable humidity-XRPD overlay in reflectance mode for Form 1 run at 25°C. [Figure 10] 5 shows an XRPD analysis in reflector mode of Example 5.1.1 performed on a Panalytical X'pert instrument. [Figure 11] 5 shows an XRPD analysis in reflector mode of Example 5.1.2 performed on a Panalytical X'pert instrument. [Figure 12] 1 shows an XRPD analysis in reflector mode of Example 5.2 performed on a Panalytical X'pert instrument. [Figure 13]1 shows an XRPD analysis in reflector mode of Example 5.2.1 performed on a Panalytical X'pert instrument. [Figure 14] 1 shows XRPD analysis in reflectance mode of Example 5.3 performed on a Panalytical X'pert instrument: (A) sample after 88 hours of filter drying, and (B) sample after subsequent tray drying. [Figure 15] 1 shows an XRPD analysis in reflector mode of Example 5.4 performed on a Panalytical X'pert instrument. [Figure 16] 1 shows an XRPD analysis in reflector mode of Example 5.4.1 performed on a Panalytical X'pert instrument. [Figure 17] Figure 1 shows XRPD overlays performed in reflectance mode on a Panalytical Empyrean instrument of Form 1, Form 2, and a slurry sample of Example 4 in water (Form 3) at various times after removing the sample from the slurry. [Figure 18] 1 shows GVS isotherm plots from multiple adsorption-desorption cycles for (A) Example 1 (Modification 1) and (B) Example 4 (Modification 3). [Figure 19] Figure 1 shows a variable humidity-XRPD overlay of Example 4 (Form 3) in reflectance mode run at 25°C. [Figure 20] Figure 1 shows XRPD overlays in reflection mode of Example 4 (Form 3) discs after compression at 20 kg, 50 kg and 100 kg. [Figure 21] 1 shows the drying profile of sulforaphane:α-cyclodextrin complex with a moisture content of 15.6% w / w when dried in a Nutsche filter dryer at 20° C. and 20 mbar vacuum with static mixing and stirring (10 rpm) according to Example 5B (Test 1). [Figure 22] 1 shows the drying profile of sulforaphane:α-cyclodextrin complex with a moisture content of 16.5% w / w when dried in a Nutsche filter dryer at 20°C, 20 mbar vacuum, and substantially constant agitation (10-20 rpm) according to Example 5B (Test 2). [Figure 23] Figure 5 shows XRPD overlays in reflectance mode of Form 3 (Example 5.3) after storage at 40°C and 75% RH for 1 month (top) and at 25°C and 60% RH for 3 months (bottom). DETAILED DESCRIPTION OF THE INVENTION

[0115] In the examples, the following abbreviations are used: DMSO - Dimethyl sulfoxide eqv - molar equivalents FaSSIF - Fasted State Simulated Intestinal Fluid GVS - Gravimetric Vapor Sorption HPLC - High Performance Liquid Chromatography KF-Karl Fischer Water Analysis NMR-Nuclear Magnetic Resonance RH - Relative Humidity RPM - Revolutions per minute RRT - Relative Retention Time TGA-Thermogravimetric analysis XRPD - X-ray powder diffraction

[0116] Equipment and Methodology XRPD XRPD diffractograms were collected on a PANalytical Empyrean diffractometer using Cu Ka radiation (45 kV, 40 mA) in transmission geometry. A 0.5° slit, a 4 mm mask, and a 0.04 rad Soller slit with a focusing mirror were used on the incident beam. A PIXcel placed on the diffracted beam 3D The detector was equipped with a receiving slit and a 0.04 rad Soller slit. The software used for data collection was X'Pert Data Collector using the X'Pert Operator Interface. Data were analyzed and presented using Diffrac Plus EVA or HighScore Plus.

[0117] Samples were prepared and analyzed in metal 96-well plates in transmission mode. X-ray transparent film was used between metal sheets on the metal well plates, and powder (approximately 1-2 mg) was used as received. The metal plate scan mode used a gonioscan axis. Details of the standard screening data collection method are as follows: ●Angle range: 2.5 to 32.0°2θ ●Step size: 0.0130°2θ ●Collection time: 12.75 seconds / step (total collection time 2.07 minutes).

[0118] Non-ambient conditions: XRPD diffractograms were collected in reflector geometry using Cu Ka radiation (45 kV, 40 mA) on a PANalytical Empyrean diffractometer. The instrument was equipped with an Anton Paar CHC plus diffractometer fitted with graphite / Kapton windows. + The stage was fitted with an air-cooled or rough vacuum pumping system using an Edwards RV3 pump in conjunction with a proUmid MHG32 modular humidity generator. A programmable divergence slit (auto mode), a 10 mm fixed incident beam mask, a Ni filter, and a 0.04 rad Soller slit were used on the incident beam. A PIXcel placed on the diffracted beam 3D The detector was equipped with a programmable anti-scatter slit (automatic mode) and a 0.04 rad Soller slit. The software used for data collection was X'Pert Data Collector, and data were analyzed and presented using Diffrac Plus EVA or Highscore Plus.

[0119] For variable humidity (VH-XRPD) experiments, samples were prepared and analyzed in an Anton Paar chrome-plated thin sample holder with a silicon wafer insert. Measurement parameters followed the standard screening data collection method (detailed above). For all variable humidity XRPD experiments, patterns were recorded every hour.

[0120] In some experiments, XRPD diffractograms were collected on a Panalytical X'pert Pro MPD X-ray diffractometer equipped with a Cu X-ray tube and a Pixcel detector system.

[0121] Additionally, XRPD diffractograms were collected on a Bruker AXS D8 diffractometer using Cu Kα radiation (40 kV, 40 mA) and a θ-2θ goniometer equipped with a Ge monochromator in a reflection geometry. The incident beam passed through a 2.0 mm anti-scatter slit, followed by a 0.2 mm anti-scatter slit and a knife edge. The diffracted beam passed through an 8.0 mm receiving slit with a 2.5° Soller slit, followed by a Lynxeye detector. The software used for data collection and analysis was Diffrac Plus XRD Commander and Diffrac Plus EVA, respectively. Samples were run under ambient conditions as flat specimens using the as-received powder. Samples were prepared on polished, zero-background (510) silicon wafers, either by gently pressing the wafer flat or by filling a cut cavity. Samples were rotated in their own plane. Standard data collection method details are as follows: angle range: 2-42°2θ; step size: 0.05°2θ; collection time: 0.5 s / step (total collection time: 6.40 min).

[0122] NMR 1 H NMR spectra were collected on a Bruker 400 MHz instrument controlled by a DRX400 console equipped with an autosampler. Samples were prepared in DMSO-d6 solvent. The automated experiment was run using the ICON-NMR configuration within Topspin software, following a standard Bruker loading experiment ( 1 H). Offline analysis was performed using an ACD Spectrus processor.

[0123] TGA TGA data were collected on a TA Instruments Discovery TGA equipped with a 25-position autosampler. Typically, 5-10 mg of each sample was placed in a tared aluminum DSC pan and heated from ambient temperature to 350 °C at 10 °C / min. A 25 ml / min nitrogen purge was maintained over the sample. The instrument control software was TRIOS, and data were analyzed using TRIOS or Universal Analysis.

[0124] GVS Adsorption isotherms were obtained using an SMS DVS Intrinsic moisture sorption analyzer controlled by DVS Intrinsic Control software. Sample temperature was maintained at 25 °C by the instrument control. Humidity was controlled by mixing dry and humid nitrogen streams at a total flow rate of 200 ml / min. Relative humidity was measured by a calibrated Rotronic probe (dynamic range 1.0–100% RH) placed near the sample. Sample weight change (mass relaxation) as a function of RH was continuously monitored with a microbalance (accuracy ±0.005 mg).

[0125] Typically, 5-30 mg of sample was placed in a tared mesh stainless steel basket under ambient conditions. Samples were loaded and removed at 40% RH and 25°C (typical room conditions). Moisture sorption isotherms were performed as outlined below (two scans per complete cycle). Standard isotherms were performed at 25°C in 10% RH intervals over the range of 0-90% RH. Typically, two cycles (four scans) were performed. Data analysis was performed in Microsoft Excel using the DVS Analysis Suite.

[0126] [Table 1]

[0127] Determination of water by KF titration The water content of each sample was measured at 150°C using an 851 Titrano Coulometer on a Metrohm 874 oven sample processor with Hydranal Coulomat AG oven reagent and a nitrogen purge. Weighed solid samples were placed in sealed sample vials. Duplicate measurements were performed using approximately 10 mg of sample per titration. Averages of these results are shown unless otherwise noted. Data collection and analysis were performed using Tiamo software.

[0128] Example 1: Formation of a Complex of Sulforaphane and α-Cyclodextrin (Form 1) To degassed α-cyclodextrin (15 g, 0.01 equiv.) in water (30 mL) was added 1-isothiocyanato-4-methylthiobutane (250 g, 1 equiv.—see WO 2013 / 179057 for the preparation of this precursor) at room temperature. The solution was cooled to below 2°C, and 31.5% aqueous HO (176 g, 1.05 equiv.) was added over 20 minutes, allowing the internal temperature to rise only to 2°C during the addition. The reaction mixture was allowed to warm to room temperature overnight with stirring. Tests showed 2.6% starting material remained, so the solution was cooled to below 2°C, and additional HO was added until tests indicated less than 1% starting material remained. The solution was filtered through a Buchner funnel to remove solids. The filtrate was used without further treatment.

[0129] α-Cyclodextrin (1523 g, 1 equiv.) was dissolved in boiling water (4 L) and cooled to 50 °C. The sulforaphane filtrate was slowly added, and the mixture was stirred at 50 °C for 1 h and then at room temperature for 24 h. The reaction mixture was then cooled and stirred at approximately 5 °C for 4 h. The resulting slurry was filtered in 1.5 L batches through a chilled Buchner funnel to keep the solution cool. The collected solid was dried overnight under vacuum on the Buchner funnel with a nitrogen stream blowing through the cake. The solid was transferred to a 5 L round-bottom Morton flask and dried under vacuum on a rotary evaporator using a 25 °C water bath for 19 h. The complex was obtained (1479 g, batch reference number LS-13-0002-S-8002) with a water content of 13.2% w / w by Karl Fischer analysis. The material had a purity of 97.5% by HPLC.

[0130] Analysis of Example 1 by XRPD (FIG. 1) showed it to be Form 1. The XRPD peak picking of Form 1 in reflector mode is summarized in Table 2.

[0131] [Table 2]

[0132] Example 1A: Complexation of sulforaphane with α-cyclodextrin by the procedure of WO 2013 / 179057 To degassed α-cyclodextrin (30 g, 0.01 eq.) in water (1 L) was added 1-isothiocyanato-4-methylthiobutane (501 g, 1 eq.) at room temperature. The solution was cooled to 0°C and degassed for 30 minutes. To the suspension was slowly added 35% aqueous H2O2 (305 ml, 1 eq.) while maintaining the internal temperature below 4°C. The reaction mixture was stirred at ice bath temperature for 8 hours and then allowed to warm to room temperature with stirring overnight. The solution was filtered to remove solids. The filtrate was refrigerated for 5 hours before use in the next step.

[0133] α-Cyclodextrin (3015 g, 1 equiv.) was dissolved in water (8 L) by heating to 55 °C. The solution was cooled to room temperature, and the sulforaphane filtrate from the previous step was added in one portion. The mixture was stirred at room temperature overnight. The reaction mixture was then cooled in an ice bath and stirred at that temperature for 3 h. The precipitated solid was filtered through a Buchner funnel and dried on the filter under vacuum overnight. The filter cake was transferred to a 10 L round-bottom flask and dried under high vacuum at room temperature over the weekend to yield a white solid (2.74 kg, 76.9% yield; batch reference number 191PAL79). The solid had a water content of 11.3% w / w by Karl Fischer analysis and a purity of 98.5% by HPLC. 1 H-NMR analysis confirmed that Example 1A was a 1:1 complex of sulforaphane:α-cyclodextrin.

[0134] Example 2: Formation of a Complex of Sulforaphane with α-Cyclodextrin (Form 2) A sample (100 mg) of Form 1 complex of sulforaphane and α-cyclodextrin was placed in a 4 mL vial and water (200 μL, 2 rel. vol.) was added to form a slurry. The vial was capped and left at room temperature overnight.

[0135] Analysis of an aliquot of the slurry of Example 2 by XRPD (FIG. 3) showed it to be a distinct form designated Form 2. The XRPD peak picking for Form 2 in reflector mode is summarized in Table 3.

[0136] [Table 3]

[0137] XRPD peak picking for Form 2 in transmission mode is summarized in Table 4.

[0138] [Table 4]

[0139] Example 3: Small-scale formation of sulforaphane and α-cyclodextrin complex (Form 3) A sample of sulforaphane:α-cyclodextrin Form 1 complex (300 mg) was weighed into a crystallization dish, covered with perforated tin foil, and placed in a vacuum oven at 40°C for 24 hours. The sample was removed from the oven and allowed to cool to ambient temperature, leaving a white solid (batch reference number: DC-1771-19-02).

[0140] Analysis of Example 3 by XRPD in reflectance mode (FIG. 4A) and transmission mode (FIG. 4B) showed it to be a new form designated Form 3. The XRPD peak picking for Form 3 in reflectance mode is summarized in Table 5.

[0141] [Table 5]

[0142] FIG. 5 shows an overlay of XRPD scans of Forms 1, 2, and 3 performed in reflectance mode.

[0143] Example 3 was reanalyzed by XRPD in reflectance mode after removal from the vacuum oven (day 0), 7, 9, and 29 days. Subsequent analyses (FIG. 6) showed that Example 3 was still present as Form 3 up to 29 days after removal from the drying oven.

[0144] 1 H-NMR (DMSO-d6, 400 MHz) confirmed that Example 3 was a 1:1 sulforaphane:α-cyclodextrin complex with no evidence of decomposition (Figure 7).

[0145] Example 3 was also analyzed by TGA at 1, 3, 7, 9, 15, and 20 days after removal from the vacuum oven and exposure to ambient conditions, as listed in Table 6.

[0146] [Table 6]

[0147] As the TGA results show, after formation, Form 3 gradually absorbs atmospheric moisture and stabilizes, with a water content of approximately 6 wt%, which corresponds to 3.5–4 water molecules per 1:1 sulforaphane:α-cyclodextrin complex.

[0148] Example 4: Formation of a Complex of Sulforaphane with α-Cyclodextrin (Mesophase Between Form 1 and Form 3) To degassed α-cyclodextrin (6 g, 0.01 eq.) in water (200 mL) was added 1-isothiocyanato-4-methylthiobutane (100 g, 1 eq.) at room temperature. The solution was cooled to 0°C, and 35% aqueous HO (60.2 mL, 1 eq.) was slowly added. The reaction mixture was allowed to warm to room temperature overnight with stirring. The reaction mixture was filtered to remove solids. The filtrate was used without further treatment.

[0149] α-Cyclodextrin (603.1 g, 1 equiv.) was dissolved in water (1.6 L) and added to the sulforaphane filtrate. The mixture was stirred overnight at room temperature. The reaction mixture was then cooled in an ice bath for 1 hour. The resulting slurry was filtered through a Buchner funnel and dried overnight under high vacuum (408 g, batch reference number 191PAL68). A sample removed from the filter cake was found to have a purity of 97.8% by HPLC and a water content of 5.8% w / w by Karl Fischer analysis. 1 H NMR analysis (D2O) confirmed the formation of a 1:1 sulforaphane:α-cyclodextrin complex.

[0150] Analysis of Example 4 by XRPD (FIGS. 8A and 8B) showed it to be an intermediate phase between Form 1 and Form 3 (major peak reflection mode at 10.49° 2θ).

[0151] The presence of such mesophases was also observed when a sample of Form 1 was exposed to variable humidity (VH) conditions at 25°C and the solid-state form was analyzed by XRPD (Figure 9). A sample of Form 1 was placed in a VH chamber set at 40% RH and 25°C. The temperature was maintained at 25°C while the RH was decreased in 10% increments to 10% RH and then increased in 10% increments to 90% RH. Upon decreasing the RH, Form 1 converted to a mesophase at 30% RH, then changed to Form 3 at 20% RH. Upon increasing the RH, the sample remained in Form 3 until the mesophase was again observed at 70% RH. The sample then rapidly transitioned from Form 1 at 80% RH to Form 2 at 90% RH.

[0152] Example 5: Large-scale formation of Form 3 complexes Example 5.1 Form 1 complex was prepared as in Example 1 to give a white solid with a purity of 98.8% by HPLC and 13.0% w / w water by KF.

[0153] Samples (10 g) were placed in crystallization dishes and dried in a vacuum oven for 72 hours at either room temperature (Example 5.1.1) or 50°C (Example 5.1.2).

[0154] Example 5.1.1 (Batch Reference A-19-0074) was found to contain 4.7% w / w water by KF and was found to be a mixture of Form 1 and primarily Form 3 by XRPD (Figure 10).

[0155] Example 5.1.2 (Batch Reference A-19-0075) was found to contain 1.8% w / w water by KF and was found by XRPD (Figure 11) to be predominantly Form 3 with very little mesophase.

[0156] Example 5.2 Sulforaphane:α-cyclodextrin complex was prepared at a 3 kg scale as in Example 1, except that after formation, the precipitated complex was filtered off through a stainless steel funnel. The cake in the funnel was dried under vacuum with an N bleed for 16 hours, then under a stream of N without vacuum for 72 hours, yielding a white solid (batch reference number 491PAL17; A-19-0098) containing 13.6% w / w water by KF. XRPD analysis of Example 5.2 (FIG. 12) showed it to be a mixture of Form 1 and Form 2.

[0157] A sample of this material (50 g) was tray dried using a freeze dryer set under vacuum for 72 hours with the tray temperature set at 21° C. (Example 5.2.1). Example 5.2.1 (batch reference A-19-0118) was found to contain 1.8% w / w water by KF and was found to be predominantly Form 3 by XRPD (FIG. 13).

[0158] Example 5.3 Sulforaphane:α-cyclodextrin complex was prepared on a 3 kg scale as in Example 1, except that after formation, the precipitated complex was filtered off through a stainless steel funnel. The cake in the funnel was dried under vacuum with limited stirring for 88 hours using a N bleed to give a white solid containing 8.5% w / w water by KF. XRPD analysis (Figure 14A; A-19-0116) showed it to be a mixture of Form 1 and Form 3.

[0159] A 1.5 kg batch of this material was further dried in trays in a drying oven at 21° C. and <1 Torr (<1.3 mbar) for 72 hours to give Example 5.3 (Batch Reference Number 491PAL18) with 2.5% w / w water by KF, and XRPD analysis (FIG. 14B; A-20-0220) showed split peaks indicating it was primarily Form 3 (10.70°2θ) with a small amount of compressed Form 3 (10.91°2θ).

[0160] Example 5.4 Sulforaphane:α-cyclodextrin complex was prepared at a 3 kg scale similar to Example 1, except that after formation, the precipitated complex was filtered off through a stainless steel funnel. The cake in the funnel was dried under vacuum with an N bleed for 72 hours, then dried under a stream of N without vacuum for 6 days. The solid was then transferred to a 20 L round-bottom flask and dried on a rotary evaporator at 25°C without stirring for 4 days (Batch Reference No. 491PAL22). Example 5.4 was found by XRPD (Figure 15) to be primarily Form 3, with some mesophase material.

[0161] A sample of this material (100 g) was tray dried using a freeze dryer set under vacuum for 72 hours with the tray temperature set at 21° C. (Example 5.4.1). Example 5.4.1 (batch reference A-19-0132) was found to contain 1.7% w / w water by KF and was found to be predominantly Form 3 by XRPD (Figure 16).

[0162] Example 5B: Form 3 Complex Formation - Filter Dryer Test Drying studies were performed on water-wet sulforaphane:α-cyclodextrin complex with a water content of 16±4% w / w by KF and isolated by filtration according to Example 1. The studies were performed on an approximately 100 g scale using a laboratory-scale Nutsche filter dryer.

[0163] The filter dryer was equipped with a temperature-controlled jacket, nitrogen supply, overhead agitation, a condenser, and a vacuum connection to the condenser pipeline. The condenser was operated at 2°C during testing to prevent water vapor from freezing the vacuum line. The jacket was set to 15°C before material loading. The wet composite was loaded through the vessel manway, the vessel was sealed, and agitation and vacuum (see Table 7 for agitation speed and vacuum setpoints) were applied to the vessel. Once the desired vacuum was achieved, the jacket temperature was increased to the desired setpoint (Table 7). A nitrogen bleed of approximately 0.45 L / min was applied to the vessel in the headspace above the loaded material, and the flow rate was monitored with an in-line flow meter. Drying was interrupted and samples were taken from the vessel to monitor drying progress by KF and / or XRPD.

[0164] [Table 7]

[0165] Overview of Test 1 Test 1 was performed with alternating periods of static drying (no agitation) and agitated drying at 10 rpm. The vacuum achieved was 19-20 mbar. Four samples were taken over a 10-hour cumulative drying period and tested for moisture content (see Figure 21 and Table H). The drying profile shows that the material initially dried rapidly, but the drying rate slowed after the moisture content fell below approximately 6% w / w. After the static drying period, more aggregated material was observed; however, these aggregates were partially broken up during the agitated drying period. The final dried material exhibited an XRPD peak at 10.70° 2θ, consistent with Form 3.

[0166] A summary of the moisture content and XRPD of the samples from Test 1 at various times during drying is shown in Table 8.

[0167] [Table 8]

[0168] Overview of Test 2 Test 2 was performed with constant agitation at 10-20 rpm. The vacuum achieved was 19-22 mbar. Six samples were taken over a cumulative drying period of 55 hours and tested for moisture content (see Figure 22). The final moisture content was 3.1% w / w, and the drying profile was consistent with Test 1, indicating that the material initially dried rapidly but then slowed down after the moisture content dropped below approximately 6% w / w. Fewer agglomerates were observed compared to Test 1, and the agglomerates decreased over time. The final dried material exhibited an XRPD peak at 10.91° 2θ, suggesting that the final material with 3.1% w / w water was overdried.

[0169] Overview of Test 3 Test 3 was performed with drying under constant agitation at 30 rpm. The vacuum achieved was 23-25 ​​mbar. A drying target of 5.0-5.5% w / w water was set. The final water content after 10 hours of drying was 5.5% w / w. Similar levels of aggregation were observed as in Test 2. The final dried material exhibited an XRPD peak at 10.76° 2θ, consistent with Form 3. This trail showed that under these conditions, a 10-hour agitated drying time was sufficient to obtain Form 3.

[0170] Overview of Test 4 Test 4 was carried out by drying with constant stirring at 30 rpm, but the temperature was increased to 30°C. The vacuum achieved was 20-22 mbar. The final moisture content after 10 hours of drying was 4.3% w / w. The final dried material exhibited an XRPD peak at 10.75° 2θ, consistent with Form 3. Similar levels of aggregation were observed as in Test 2.

[0171] Test 5 Overview Test 5 was carried out by drying at 30 rpm and 20°C with constant stirring, but the vacuum was reduced to 50-70 mbar. The final moisture content after 9 hours of drying was 5.3% w / w. The final dried material exhibited an XRPD peak at 10.71° 2θ, consistent with Form 3. Similar levels of aggregation were observed as in Test 2.

[0172] A sample taken after 5 hours of drying had a water content of 6.5% w / w and a major XRPD peak at 10.54°2θ. This suggests that a drying time of at least 9 hours would be required to obtain material with the target water content of 5.0-5.5% w / w if the vacuum was reduced. This drying time is consistent with Tests 1 and 3, which used a 20 mbar vacuum, and it was concluded that a reduction in vacuum strength from 20 mbar to 50-70 mbar could be tolerated without affecting drying efficiency.

[0173] Test 6 Overview Test 6 was run using a different feed material with a higher moisture content. The material from Test 1 was slurried in 2.57 volumes of water in a filter dryer vessel at 5° C. for 3.5 hours. The slurry cake was blown down at a pressure of 0.5 to 0.75 bar G to give a material with a moisture content of 26.4% w / w. This material was blown down under nitrogen for a further hour and then held overnight at a jacket temperature of 0° C. The blown down cake showed evidence of cracking and had a moisture content of 29.5% w / w.

[0174] This material, with a high moisture content of 25-30% w / w, was then dried at 20°C for 9 hours under a vacuum intensity of 27-28 mbar. The cake in the filter dryer was smaller in height than that observed in previous tests, resulting in insufficient mixing. Initial agitation at 30 rpm showed mixing within the cake similar to that observed previously, but balling (golf-ball-sized clumps) was observed within 15 minutes of agitation initiation. At this point, agitation was stopped, the agitator direction was changed from "mixing" to "smoothing," and agitation was continued at 10 rpm to break up the balling. Once this was achieved, the agitation direction was changed back to "mixing," and agitation continued at 10 rpm. After 1 hour, when no more balling was observed, the agitation speed was increased to 30 rpm. No significant agglomeration was observed throughout the remainder of the drying test.

[0175] The final moisture content after 9 hours of drying was 5.1% w / w. The final dried material exhibited an XRPD peak at 10.55° 2θ consistent with Form 3. However, the overall XRPD pattern showed reduced intensity and broad peaks likely associated with the presence of mixed phases.

[0176] Filter Drying Test - Evaluation of Flocculation The degree of agglomeration in each of the filter drying tests 2-6 was assessed by passing a portion of each discharged dried material through a 2000 micron sieve and quantifying the excess residue relative to the input amount. The remaining agglomerates were analyzed by XRPD in the same manner as the test samples, with the modification of using a zero-background silicon holder with a cavity due to the sample volume. The results are summarized in Table 9 below.

[0177] [Table 9]

[0178] It can be seen that while the dried samples typically contain less than 5% w / w of agglomerates with particle sizes greater than 2000 μm, the material from Test 6 had a much higher level of agglomerates at 23.1% w / w. The XRPD main peak values ​​in Table I also show that conversion to Form 3 is less complete for the agglomerated material than for the bulk fine material.

[0179] Filter Drying Test - Conclusion Optimal drying conditions comprise agitated drying (preferably continuous agitated drying) to reduce drying time and agglomeration to ensure reproducible production of Form 3.

[0180] A suitable nitrogen bleed and a vacuum of at least 70 mbar combined with agitation drying allows efficient drying to Form 3 within a time frame of less than 10 hours.

[0181] A drying target of 5.0-5.5% w / w water was adequate to obtain Form 3 material (major XRPD peak in reflection mode at 10.7±0.2°2θ).

[0182] It is not necessary to carry out agitation drying at temperatures above 20°C, although temperatures up to 30°C may be tolerated while minimizing degradation of the sulforaphane:α-cyclodextrin complex.

[0183] Using input materials with too high a moisture content (greater than approximately 25% w / w water) will adversely affect drying efficiency and the physical properties of the dried material, necessitating an additional non-agitated drying step.

[0184] Example 6: Testing of Form 3 aqueous operation Example 3 - Form 3 (100 mg) was placed in a 4 ml vial and HO (200 μl, 2 volumes) was added. The sample was stoppered and left overnight at ambient temperature. A spatula-full of the slurry was placed in a well and analyzed by XRPD in transmission mode. XRPD analysis was repeated on the same sample in the well every 5 minutes for 36 minutes. An XRPD was also recorded after 4 hours. The XRPD is shown in Figure 17. Form 3 converted to Form 2 when slurried in water, as seen by the XRPD patterns recorded after 14 and 36 minutes. After standing at ambient conditions for an additional 4 hours, the sample converted from Form 2 to Form 1.

[0185] Exposure to various humidity levels GVS Samples of Example 1 (Form 1) and Example 3 (Form 3) were analyzed for their adsorption / desorption isotherms by GVS. Figure 18A shows the GVS data for the Form 1 sample. The sample contained approximately 8% w / w water at starting conditions of 40% RH and 25°C. When the RH was increased to 90%, the sample took up an additional 4-5% w / w water, resulting in a total water content of 12.9% w / w. There was a noticeable hysteresis between 70% and 30% RH. After the first adsorption / desorption cycle, Form 1 was converted to Form 3, as confirmed by XRPD. Thus, the second adsorption profile seen in Figure 18A is that of Form 3. Figure 18B shows the GVS data for the Form 3 sample. The sample contained approximately 3-4% w / w water at starting conditions of 40% RH and 25°C. Increasing the RH to 90% caused the sample to take up an additional 7–8% w / w of water, resulting in a total water content of 11.1% w / w. Significant water uptake, equivalent to approximately 5% w / w, occurred during the first adsorption cycle at RH between 60–70%. XRPD analysis confirmed that the crystalline complex remained in Form 3 after GVS.

[0186] These results indicate that Form 3 has better control over water content than Form 1. The GVS data show that between 0% and 60% RH, Form 3 only takes up about 4% w / w of water, whereas at 60% RH, Form 1 takes up up to 8-9% w / w of water.

[0187] VH-XRPD Variable humidity (VH) XRPD experiments were performed to investigate the existence of Form 3 under different RH conditions. The VH-XRPD humidity profile for the sample from Example 3 (Form 3) is shown in Figure 19. The sample was loaded into a VH chamber and set to 40% RH and 25°C. The RH was then increased in 10% increments up to 90% RH, held at each RH increment for up to 8 hours, and then gradually decreased back to 40% RH while maintaining the temperature at 25°C. Form 3 persisted up to 60% RH, but a mesophase formed upon increasing the RH to 70% RH, resulting in Form 2 at 80% RH. The sample remained stable as Form 2 for 8 hours at 80% RH. At 90% RH, the sample deliquesced after 2 hours of Form 2 in the humidity chamber. Reducing the VH to 80% RH reverted Form 2 to a highly crystalline material. At 70% RH, Form 2 converted to Form 1, which persisted until the sample was removed from the 40% RH humidity chamber.

[0188] VH-XRPD experiments show that Form 3 is stable over a wide RH range. From 0% to 60% RH, Form 3 is stable at 25°C. Form 3 begins to convert to another form only after 70% RH is reached. Thus, Form 3 offers the advantage of improved handling over a wide RH range and better control of the crystalline complex form during storage over Form 1. Table 10 summarizes the three forms of crystalline 1:1 sulforaphane:α-cyclodextrin complex identified and their stability at 25°C:

[0189] [Table 10]

[0190] Compression Test A 15 mg sample of Example 3 (Form 3) was compressed into a 3 mm concave disk at pressures of 20 kg, 50 kg, and 100 kg for 2 minutes each. 1 H-NMR showed no decomposition of the compressed sample. Figure 20 shows an XRPD overlay of the compressed sample, confirming that the sample still exists as Form 3, although there was a slight loss in intensity of the major Bragg peaks.

[0191] Consolidation Test Samples of Form 1 and Form 3 were subjected to Haeckel analysis to determine the yield stress of the material during compression under controlled conditions. A known weight of material was compressed in a 10 mm diameter die with a flat-faced punch moving at a set speed. The die was lubricated with magnesium stearate in acetone. Punch force was measured frequently and accurately, while punch displacement was used to calculate powder volume. Yield pressures were calculated at low (0.1 mm / s) and high (300 mm / s) punch speeds to assess the time-dependent component to material deformation. The true density of the material was determined by helium pycnometer (Micromeritics AccuPyc II 1340) using purge and run-fill pressures of 19.5 psig and an equilibration rate of 0.02 psig. Tests were performed in duplicate. Temperature and humidity were monitored at intervals during testing.

[0192] The data was analyzed by a consolidation analysis software program to generate a value for yield pressure (Py) using the Haeckel equation: ln(1 / 1-D)=kP+A where D = relative density of the compact form; P = applied pressure; and K = slope of the line in the linear region [Heckel, Trans. Metall. Soc. AIME 221 (1961) 1001-1008].

[0193] Strain rate sensitivity (SRS) can be calculated to determine whether the deformation characteristics of a material change with the rate of applied force. The yield stress during high-speed compression is compared to the yield stress during low-speed compression using the following equation [Roberts and Roe, Chem. Eng. Sci. 42 (1987) p. 903]: %SRS=100x[(Py Fast-Py Slow) / Py Slow]

[0194] Loss on drying of the samples was performed at the beginning and end of the compression test to monitor moisture absorption during the test. Sample sizes of 2.0 g ± 0.2 g were spread onto aluminum trays and dried at 105 °C until stable using a Mettler HB43-S halogen moisture meter.

[0195] The results of the compaction test are shown in Table 11.

[0196] [Table 11]

[0197] The compaction results show that morphology 1 is classified as soft ductile, whereas morphology 3 is classified as medium hard brittle / ductile.

[0198] The yield pressure for Form 1 at low speed was 76 MPa, indicating that low to moderate force was required for the material to deform. The yield pressure for Form 3 at low speed was 109 MPa, indicating that greater force was required for the material to deform.

[0199] Strain rate sensitivity (SRS) measures the change in compaction behavior with production compaction speed. At high speed, the yield pressure increased to 89 MPa for Configuration 1 and 140 MPa for Configuration 3. The results show an SRS of +16.5% (Configuration 1) and +29.1% (Configuration 3). Both configurations exhibit differences in deformation with increasing speed, which may lead to differences in compaction upon scale-up. At higher speeds, increased force is required to achieve the same level of compaction. There was no evidence of adhesion to the punch or die walls for either configuration.

[0200] Compaction results revealed that Form 3 has a moderate yield pressure in the ideal range for tablets (80-120 MPa).

[0201] Storage at high humidity and / or high temperature A sample of Example 3 (Form 3) was stored at 25° C. and 97% RH for 7 days. 1H-NMR did not reveal any decomposition, but XRPD showed that Form 3 was converted to Form 1 at this ultra-high humidity level.

[0202] A sample of Example 3 (Form 3) was stored at 40° C. and 75% RH for 7 days. 1 Analysis by H-NMR showed that some decomposition occurred under these conditions. Similar decomposition was observed for Form 1 under these conditions.

[0203] Example 7: Preliminary stability study of Form 3 Analysis of samples of the sulforaphane:α-cyclodextrin complex prepared herein was carried out by HPLC according to the following method: ● Column - Phenomenex Gemini C18, 5 μm, 110 Å, 250 × 4.6 mm; ●Mobile phase A: water (+0.1%TFA); ● Mobile phase B: acetonitrile (+0.1% TFA); ●Flow rate: 1.5mL / min; ●Injection volume: 10μL; ●Column temperature: 25℃; ●Running time: 40 minutes; ● Sample preparation - Approximately 320 mg ± 30 mg of sample was accurately weighed and transferred to a 25 mL volumetric flask, and approximately 15 mL of diluent (70% water, 30% acetonitrile, 0.1% TFA) was added and mixed until dissolved. Diluted to volume with diluent and mixed. 3-4 mL was transferred to a syringe and filtered using a 0.45 PTFE filter, discarding the first 1-2 mL, and filtered into an autosampler vial, which was sealed with a PTFE-lined cap (prepared in duplicate); HPLC gradient according to Table 12

[0204] [Table 12]

[0205] Sulforaphane and certain related impurity peaks are summarized in Table 13. Impurity characterization was performed using LC-MS and1 The results were analyzed by H-NMR. Relative retention times (RRT) are based on the HPLC method described above.

[0206] [Table 13]

[0207] Samples of Form 1 (Batch Reference No. LS19-0004-S-8002), Form 3 prepared by minimal agitation drying (Example 5.3; Batch Reference No. 491-PAL-18), and Form 3 prepared by continuous agitation filter drying (Example 5B-Test 5; Batch Reference No. 00173-005) were stored in 60 ml Duma® bottles with 45 mm Duma® caps at 5° C. (Table 14) and 25° C. / 60% RH (Table 15) for 8 weeks and the samples were analyzed by HPLC according to the method described above.

[0208] [Table 14]

[0209] [Table 15]

[0210] It should be noted that the "initial" time point is not immediately after synthesis, but immediately after the initiation of the respective stability study (e.g., Example 5B-Test 5 samples were refrigerated after synthesis for 5-6 months before the initiation of stability testing). Therefore, this preliminary (8-week) stability study is not a fully comparative study of samples prepared at the same time and stored under comparable conditions.

[0211] Example 7B: Moisture Content of Stability Samples Samples of Form 1 (Batch Reference No. LS19-0004-S-8002), Form 3 prepared by minimal agitation drying (Example 5·3; Batch Reference No. 491-PAL-18), and Form 3 prepared by continuous agitation filter drying (Example 5B-Test 5; Batch Reference No. 00173-005) were analyzed for moisture content by KF at the start of the stability study (initial) and after 2, 4, and 8 weeks of storage at 25°C / 60% RH in 60 ml Duma® bottles (without desiccant) with 45 mm Duma® caps; see Table 16.

[0212] [Table 16]

[0213] In another study, a sample of Form 3 (Example 5.3; Batch Reference No. 491-PAL-18) having an initial moisture content of 2.3% w / w by KF was tested for moisture content at various time points after double-bagged storage in HDPE bottles under various conditions, as shown in Table 17.

[0214] [Table 17]

[0215] In general, overdried Form 3 (e.g., less than 4% w / w water) tends to take up water, but the data above show that even when Form 3 material is exposed to high levels of humidity for extended periods of time, such as up to 6 months, the water content does not increase significantly beyond about 5-6% w / w.

[0216] Figure 23 shows an overlay of the XRPD plots of the 1-month (40°C / 75%RH) and 3-month (25°C / 60%RH) samples obtained in this study, showing that the major peak is consistent with Form 3 (10.7±0.2°2θ).

[0217] Example 8: Chemical stability of Forms 1 and 3 in biorelevant media Samples of Form 1 (Batch Reference L17-0004-S-8002) and Form 3 (Example 5.4.1 (Batch Reference 491PAL22; A-19-0132)) in 0.1 M HCl and FaSSIF medium (1.2 mg / ml) were stored at ambient and 37° C. for up to 7 days. Chemical stability was assessed by measuring the concentration of the parent compound by HPLC in triplicate on days 0, 3, and 7.

[0218] [Table 18]

[0219] The data in Table 18 show that while some parental loss was observed in acidic media, there was approximately 1.5-fold more parental loss over the same period in FaSSIF. Little difference was observed between Forms 1 and 3 in terms of parental loss rate.

[0220] Example 9: Equilibrium solubility of Form 1 and Form 3 in biorelevant media Equilibrium solubility in 0.1 M HCl and FaSSIF media at 37°C was performed using the shake flask method for samples of Form 1 (Batch Reference L17-0004-S-8002) and Form 3 (Example 5.4.1 (Batch Reference 491PAL22; A-19-0132)). Samples were taken after 4 and 24 hours and centrifuged at 15,000 rpm for 5 minutes. The supernatant was removed and diluted with an appropriate solvent before being subjected to HPLC analysis of the parent concentration. Samples made in FaSSIF produced atypical HPLC data and variable results.

[0221] [Table 19]

[0222] Although some variation in the equilibrium solubility of Forms 1 and 3 was observed, the data in Table 19 show that both forms were highly soluble and comparable (within experimental error) in both 0.1 M HCl and FaSSIF media. Solubility was higher in acidic media versus FaSSIF media for both forms.

[0223] Example 10: Kinetic solubility of Form 1 and Form 3 in biorelevant media Small-scale dynamic solubility studies were performed at nominal concentrations of 200-400 mg / ml to ensure that solid material remained at the end of the experiment for XRPD analysis. Approximately 100 mg of sample—Form 1 (Batch Reference L17-0004-S-8002) or Form 3 (Example 5.4.1 (Batch Reference 491PAL22; A-19-0132))—was weighed into a 1 ml vial, and 250 μl of 0.1 M HCl was added to the vial for the 15 and 30 minute time points, and 500 μl of a 1:1 mixture of 0.1 M HCl and FaSSIF was added to the vial for the 60 and 180 minute time points. The vial was agitated and maintained at 37°C.

[0224] Tests were performed in duplicate. Each sample was centrifuged to remove suspended solids. An aliquot of the supernatant was transferred to a second vial for further centrifugation, then transferred to an LC vial, diluted with an appropriate solvent, and analyzed for parent compound by HPLC. The remaining solids were analyzed by XRPD.

[0225] [Table 20]

[0226] As shown by the data in Table 20, both Form 1 and Form 3 dissolved rapidly in 0.1 M HCl, achieving approximately 300 mg / ml by the 15 minute time point. The pH shift, representing transition from the stomach to the intestinal environment, resulted in a decrease in the parent concentration to approximately 180 mg / ml at the 60 and 180 minute time points. XRPD analysis of the residual solids indicates that Form 1, or a mixture of Forms 1 and 3, was observed at the end of the study.

Claims

1. A crystalline complex of sulforaphane and α-cyclodextrin having a water content of less than 8% w / w, wherein the crystalline form of the complex is stable at 25°C and between 0% and 60% relative humidity.

2. 2. The crystalline complex of claim 1, having a water content of less than 6% w / w.

3. The molar ratio of sulforaphane to α-cyclodextrin in the complex is i. in the range of 0.9:1 to 1.1:1; ii. 1:1; The crystalline composite of claim 1 or claim 2.

4. The crystalline form of the complex is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflection mode; optionally: It may be further characterized by additional XRPD peaks at 8.1 and 16.1±0.2 degrees 2θ when measured in reflectance mode Crystalline complex of sulforaphane and α-cyclodextrin (Form 3).

5. A crystalline complex according to any one of claims 1 to 4, wherein the complex comprises less than 5% w / w of particles greater than 2000 μm in diameter.

6. a) providing a complex of sulforaphane and α-cyclodextrin; b) drying the composite from step a) while stirring the composite at a pressure of less than 200 mbar until the moisture content of the composite is less than 6% w / w; 6. A method for forming a crystalline complex of sulforaphane and α-cyclodextrin according to any one of claims 1 to 5, comprising:

7. one or more of the following steps: i. the drying of step b) is carried out at a pressure of less than 100 mbar; ii. the drying of step b) is carried out under a flow of inert gas, which may optionally be nitrogen; iii. The drying of step b) is carried out with constant agitation; iv. the agitation in step b) is provided at a speed greater than 5 rpm; v. the drying of step b) is carried out for at least 5 hours; vi. the drying of step b) is carried out at a temperature between 10°C and 30°C; vii. the complex provided in step a) has a water content of 10-25% w / w, and / or viii. Drying step b) is carried out until the moisture content of the complex is 4-6% w / w. The method of claim 6, wherein:

8. A solid pharmaceutical composition comprising an effective amount of the crystalline complex of any one of claims 1 to 5, and optionally at least one pharmaceutically acceptable excipient.

9. 10. The solid pharmaceutical composition of claim 8, wherein greater than 50% of the crystalline complex of sulforaphane and α-cyclodextrin is characterized by XRPD peaks at 5.3 and 10.7±0.2 degrees 2θ when measured in reflectance mode.

10. 10. The solid pharmaceutical composition of claim 8 or 9, wherein the composition is formulated as a tablet or capsule.

11. i. a disease or disorder mediated by Nrf2 or STAT3; ii. Cancer, subarachnoid hemorrhage, intracerebral hemorrhage, ischemic stroke, delayed cerebral ischemia, atherosclerosis, middle cerebral artery infarction, pulmonary arterial hypertension, Alport syndrome, nonalcoholic steatohepatitis, focal segmental glomerulosclerosis, Huntington's disease, Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis, cirrhosis, nonalcoholic fatty liver disease, hepatitis, acute kidney injury, sepsis, chronic kidney disease, systemic lupus erythematosus nephritis, emphysema, pulmonary fibrosis , chronic obstructive pulmonary disease, asthma, inflammatory lung disease, lymphocytic interstitial pneumonia, splenomegaly, type I diabetes, type II diabetes, diabetic nephropathy, diabetic retinopathy, diabetic cardiomyopathy, hyperglycemia, arthritis, scleroderma, atopic dermatitis, lymphadenopathy, alopecia, hyperthyroidism, esophageal achalasia, thrombocytopenia, neutropenia, autoimmune hemolytic anemia, frontotemporal dementia, autism spectrum disorder, multiple sclerosis or Friedreich's ataxia; iii. breast cancer; or iv. ER+ or HER2- metastatic breast cancer A pharmaceutical composition according to any one of claims 8 to 10 for use in the treatment of

12. 11. The conjugate of any one of claims 1 to 5 or the pharmaceutical composition of any one of claims 8 to 10 in combination with one or more additional therapeutic agents, wherein the additional therapeutic agents comprise an aromatase inhibitor, tamoxifen, exemestane, fulvestrant, an oral SERD or CDK4 / 6 inhibitor.

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