Crystal form

Abstract

The present invention relates to a compound of formula (I): The present invention further relates to novel crystalline forms of the compound of formula (I) or its adduct, and to pharmaceutical compositions comprising the same, methods for preparing pharmaceutical compositions, and uses and medical treatments using the novel crystalline forms.

Description

[Technical field]

[0001] Technical Field The present invention relates generally to novel crystalline forms of certain histone deacetylase inhibitors and their adducts. The present invention also relates to pharmaceutical compositions comprising the novel crystalline forms, methods for preparing the novel crystalline forms and pharmaceutical compositions, and their use in treating diseases, such as proliferative or autoimmune diseases. [Background technology]

[0002] Background Histone deacetylases (HDACs) Histone deacetylases (HDACs) are important regulators of cancer (see PA Marks et al., Nature Rev. Cancer, 2001, 1, 194; JE Bolden et al., Nature Rev. Drug Discov., 2006, 5, 769; P. Gallinari et al., Cell Res., 2007, 17, 195; KB Glaser, Biochem. Pharmacol., 2007, 74, 659; L. Pan et al., Cell. Mol. Immunol., 2007, 4, 337; M. Haberland et al., Nature Rev. Genetics, 2009, 10, 32; Y. Zhang et al., Curr. Med. Chem., 2008, 15, 2840; S. Ropero and M. Esteller, Mol. Oncol., 2009, 16, 2840). 2007, 1, 19) and other diseases, such as those related to the central nervous system, autoimmune diseases (see AG Kazantsev and LM Thompson Nature Rev. Drug Discov. 2006, 7, 854).

[0003] Several families of HDAC inhibitors (HDACi) have been designed and their general structures can be found in different reviews (see A. Villar-Garea and M. Esteller Int. J. Cancer 2004, 112, 171; TA Miller et al. J. Med. Chem. 2003, 46, 5097; T. Suzuki and N. Miyata Curr. Med. Chem. 2005, 12, 2867; M. Paris et al. J. Med. Chem. 2008, 51, 1505). The general structure of these inhibitors consists of a ring structure, a spacer and a chelating group that can bind to the Zn(II) cation in the active center of different HDAC isoforms belonging to class I (HDAC1, HDAC2, HDAC3 and HDAC8), class II (HDAC4, HDAC5, HDAC6, HDAC7, HDAC9 and HDAC10) and class IV (HDAC11).

[0004] The mechanism of action of HDAC inhibitors is explained by their antagonistic properties against histone deacetylases, which are involved in the control of processes related to apoptosis, cell proliferation, tumor progression, cancer metastasis, cell adhesion, etc. These properties are achieved by preventing HDACs from binding to their natural ligands, which are cytoplasmic proteins such as histones or tubulin, as well as by inhibiting their normal catalytic activation, i.e., deacetylation of ε-N-acetyl lysine residues present in these proteins.

[0005] Despite having a similar mode of inhibition, occasionally some selectivity has been observed in the inhibition of different HDAC isoforms (see JC Wong et al., J. Am. Chem. Soc. 2003, 125, 5586; G. Estiu et al., J. Med. Chem., 2008, 51, 2898). The selectivity mentioned is of therapeutic interest (see KV Butler and AP Kozikowski, Curr. Pharm. Design, 2008, 14, 505; TC Karagiannis and A. El-Osta, Leukemia, 2007, 21, 61).

[0006] HDAC inhibitors One important class of HDAC inhibitors are trisubstituted pyrrole derivatives linked to chelating groups via aromatic and heteroaromatic groups, as described, for example, in WO 2011 / 039353. These compounds have been shown to be effective in the treatment of cancer (see WO 2011 / 039353).

[0007] Furthermore, these compounds have been shown to be effective in treating several autoimmune diseases, for example, they have been shown to be effective in animal models of autoimmune hepatitis and autoimmune encephalomyelitis (see WO2018 / 087082).

[0008] A particularly promising compound is 3-(3-furyl)-N-{4-[(hydroxyamino)carbonyl]benzyl}-5-(4-hydroxyphenyl)-1H-pyrrole-2-carboxamide (referred to herein as QTX125).

[0009] [ka]

[0010] QTX125 is a highly selective and potent HDAC6 inhibitor. It has shown high antitumor efficacy in xenograft murine models of mantle cell lymphoma (see Perez-Salvia, M. et al Haematologica 2018; 103:e540), lung cancer and pancreatic cancer. QTX125 has also shown high efficacy in two different mouse models of multiple sclerosis (see WO 2018 / 087082).

[0011] However, hydroxamic acids such as QTX125 are known to have very low solubility in water (see Patre, S. et al., International Conference on Environment and BioScience IPCBEE, 2011, vol. 21), and high pH values ​​are usually required to dissolve QTX125 in aqueous solutions. QTX125 has also been demonstrated to be physically and chemically unstable in solution.

[0012] As a result, there remains a need in the art to provide novel forms of QTX125 that are particularly useful, but not limited to, in pharmaceutical formulations, particularly novel forms of QTX125 that contain high concentrations of QTX125 at physiological pH, and that are stable and have low toxicity are particularly desirable.

[0013] Several patents and publications are cited herein in order to more fully describe and disclose the invention and the state of the art to which it pertains. Full citations for these documents are provided herein. Each of these documents is incorporated herein by reference in its entirety into this disclosure. Summary of the Invention

[0014] Summary of the Invention The present inventors have developed crystalline forms of QTX125 and its adducts which help to solve the practical problems mentioned above.

[0015] In one aspect, the present invention provides a compound of formula I:

[0016] [ka]

[0017] or a crystalline form of an adduct thereof, characterized by a powder X-ray diffraction (PXRD) pattern having peaks at 2θ=20.4°, 21.8°, 22.0°, 22.7° and 23.9° (±0.3° 2θ).

[0018] The compound of formula I is also referred to herein as QTX125. The compound of formula I (QTX125) is 3-(3-furyl)-N-{4-[(hydroxyamino)carbonyl]benzyl}-5-(4-hydroxyphenyl)-1H-pyrrole-2-carboxamide.

[0019] Another aspect of the present invention relates to pharmaceutical compositions comprising a crystalline form of the compound of formula I of the present invention or an adduct thereof.

[0020] Another aspect of the present invention relates to an in vitro complex comprising a crystalline form of the compound of formula I of the present invention or an adduct thereof.

[0021] Further aspects of the present invention relate to processes for the preparation of crystalline forms of the compounds of formula I of the present invention or their adducts, and to the crystalline forms of the compounds of formula I of the present invention or their adducts obtained from these processes.

[0022] Further aspects of the present invention relate to methods for preparing pharmaceutical compositions comprising a crystalline form of a compound of formula I of the present invention or an adduct thereof, and to pharmaceutical compositions obtainable by such methods.

[0023] Another aspect of the present invention relates to a crystalline form of the compound of formula I of the present invention or an adduct thereof, for use as a medicament.

[0024] Another aspect of the present invention relates to the use of a crystalline form of the compound of formula I of the present invention or its adduct in the preparation of a medicament.

[0025] According to another aspect, the present invention relates to a method for treating a mammal comprising administering to a patient in need of such treatment a therapeutically effective amount of at least one crystalline form of the compound of formula I of the present invention or an adduct thereof.

[0026] According to certain embodiments, the crystalline forms of the compound of formula (I) of the present invention or its adduct are useful in the treatment of various types of cancers by limiting tumor growth or other processes that halt the development of primary or metastatic tumors through the inhibition of certain histone deacetylases.

[0027] According to a particular embodiment, the adduct of the compound of formula I of the invention is an adduct with lysine, in particular a 1:2 adduct with L-lysine.

[0028] BRIEF DESCRIPTION OF THE DRAWINGS BRIEF DESCRIPTION OF THE DRAWINGS Embodiments and experiments illustrating the principles of the present invention will now be discussed with reference to the accompanying drawings in which: [Brief description of the drawings]

[0029] [Figure 1] FIG. 1A shows a thermogravimetric analysis (TGA) graph of crude QTX125. Delta Y displays 8.252%. FIG. 1B shows an overlay of graphs generated from differential scanning calorimetry (DSC) (dark grey) and TGA (light grey) of crude QTX125. Delta Y displays 8.252%. FIG. 1C shows a powder X-ray diffraction (PXRD) pattern of crude QTX125. [Diagram 2]Figure 2A shows the PXRD pattern of QTX125 Form2 isolated by extractive purification and water slurry. Figure 2B shows a plot generated from DSC of Form2 alone. The displayed onset is 213.97°C, the displayed peak is 221.72°C, the peak height is -7.1369mW, the area is -356.666mJ, and the delta H is -178.3332J / g. Figure 2C shows a plot generated from DSC of crude QTX125 (dark grey, top) overlaid with Form2 (light grey, bottom). The displayed onset, peak, peak height, area, and delta H values ​​are as shown in Figure 2B. For crude QTX125, the displayed corresponding peak and peak height values ​​are 210.55°C and 13.8539mW. Also shown is 22.6070 mW at 181.08 °C, which corresponds to a minor endothermic peak in crude QTX125. Figure 2D shows an overlay of the TGA (dark grey) and DSC (light grey) graphs of Form2. The displayed delta Y is 12.255%. [Diagram 3] Figure 3A shows an overlay of the PXRD pattern of Form 2 isolated by crystallization (top) and the PXRD pattern of Form 2 isolated by extractive purification and water slurry (bottom). Figure 3B shows a graph generated from DSC of Form 2 isolated by crystallization. Shown is an onset at 234.28°C, a peak at 237.25°C, a peak height of -30.1217mW, an area of ​​-545.835mJ, and a delta H of -227.4312J / g. Also shown is -11.8407mW at 233.20°C, which corresponds to a minor endothermic event. [Figure 4]FIG. 4A shows the PXRD patterns of Form2 isolated from (i) the first scale-up crystallization (top); (ii) the trial crystallization (middle); and crude QTX125 (bottom). FIG. 4B shows graphs generated from DSC of Form2 isolated from (i) the first scale-up reaction (lightest gray, bottom); and (ii) the trial crystallization (medium gray, middle); and crude QTX125 (dark gray, top). The peaks displayed are (i) 235.88° C. (peak height is −4.8278 mW); (ii) 236.84° C. (peak height is 0.5430 mW); and (iii) 210.31° C. (peak height is 13.8495 mW), respectively. Also displayed are 22.66056 mW- at 180.53 °C, which corresponds to a minor endothermic peak for crude QTX125, and 19.3510 mW- at 229.49 °C and 229.89 °C (peak height 18.1257 mW), which correspond to minor endothermic events for Form2. Figure 4C shows a graph generated from the DSC (light grey) and TGA (dark grey, top) of Form2 isolated from the first scale-up reaction; and an overlay of the TGA graph (light grey, bottom) of QTX125 Form2 isolated via trial crystallization. For the first scale-up of Form2, the displayed delta Y is 8.500% and for the trial crystallization of Form2, it is 9.229%. The displayed peak is 235.88 °C (peak height -4.8292 mW). Also reported is 18.1258 mW at 229.89° C., which corresponds to a minor endothermic event in Form 2. [Diagram 5] Figure 5A shows the PXRD pattern of Form2 isolated from the first scale-up (bottom) and the PXRD pattern of Form2 isolated from the second scale-up (top). Figure 5B shows an overlay of the DSC (light grey) and TGA (dark grey) patterns of Form2 isolated from the second scale-up reaction. The indicated delta Y is 8.384% and the indicated peak is at 238.66°C (peak height is -1.1993mW). Also shown is 19.2562mW at 228.78°C, which corresponds to a minor endothermic event for Form2. [Figure 6] Figure 6A shows the solubility profile of QTX125 Form2 in six aqueous solutions discussed in Example 5 (briefly: phosphate buffer pH 3.5; phosphate buffer pH 6.5; acetate buffer pH 4.5; citrophosphate buffer pH 4.5; 0.9% w / v NaCl, and water). Figure 6B shows the profile expansion of the solubility profile of QTX125 Form2 in five of these solutions (per Figure 6A, excluding water). [Figure 7] Figure 7A shows the solubility profile of the QTX125 1:2 L-lysine adduct in six aqueous buffers (same as those displayed in Figure 6A), and Figure 7B shows a profile expansion of the solubility profile of the adduct in four of these buffers (from Figure 7A except for 0.9% w / v NaCl and water). [Figure 8] Solubility profiles of two QTX125 (Form 2, black, and 1:2 L-lysine adduct, grey) entities in deionized water. [Figure 9] PXRD pattern of crystals of QTX125 1:2 L-lysine adduct. [Figure 10] Overlay graphs made from TGA and DSC of crystals of QTX125 1:2 L-lysine adduct. (a) shows TGA delta Y=0.850%; (b) shows TGA delta Y=0.593%; (c) shows a DSC peak of 21.2157 mW at 95.38°C; (d) shows a TGA delta Y=4.685%; (e) shows a DSC peak of 22.7664 mW at 155.85°C; (f) shows a DSC peak of 22.3195 mW at 167.51°C; (g) shows a DSC peak of 21.7741 mW at 184.40°C.

[0030] Detailed Description definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs.Methods and materials are described herein for use in this disclosure, but other suitable methods and materials known in the art can also be used.Materials, methods, and examples are illustrative only and are not intended to be limiting.All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety.In case of discrepancy, the present specification, including definitions, will prevail.

[0031] The term “C x -C y "Alkyl" refers to a straight or branched hydrocarbon chain consisting of carbon and hydrogen atoms, containing no unsaturation, having from x to y carbon atoms. For example, the term "C1-C4 alkyl" refers to a straight or branched hydrocarbon chain consisting of carbon and hydrogen atoms, containing no unsaturation, having from 1 to 4 carbon atoms, preferably from 1 to 3 carbon atoms ("C1-C3 alkyl"), attached to the remainder of the molecule via a single bond, examples of which include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, etc.

[0032] The term "about" preceding a stated value indicates that the value may have an uncertainty of ±20%, preferably ±10%, ±5%, ±2%, ±1% of the stated value.

[0033] The term "room temperature" refers to the ambient temperature of a typical laboratory, typically between 20°C and 30°C, preferably around 25°C, at atmospheric pressure.

[0034] The term "dry" refers to an ingredient, such as a crystalline form or composition, that has been subjected to drying. Optionally, it may refer to a solid material having a residual moisture content of less than 10%, preferably less than 8%, preferably less than 5%, preferably from about 0.1% to about 5%. Residual moisture content may be determined using Karl Fischer titration.

[0035] The term "injection" refers to any form of injection known to those skilled in the art, such as subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid, and intrasternal. Injection can also refer to the process of infusion (e.g., continuous administration) as well as bolus (discontinuous) administration.

[0036] The term "pharmaceutical acceptable salt" refers to a salt that, when administered to a recipient, can provide (directly or indirectly) a compound described herein. "Pharmaceutically acceptable" preferably refers to compositions and molecular entities that are physiologically acceptable and do not normally produce allergic reactions or similar undesirable reactions, such as stomach upset, dizziness, or the like, when administered to humans or animals. Preferably, the term "pharmaceutical acceptable" means approved by a state or federal regulatory agency or included in the United States Pharmacopeia or other generally recognized pharmacopoeias for use in animals, particularly humans.

[0037] The term "adduct" refers to the product of the direct addition of two or more different molecules, resulting in a single reaction product that contains every atom of every component. For example, the adduct of QTX125 and L-lysine, discussed further herein, is believed to be produced by the interaction between L-lysine and QTX125.

[0038] The adducts can be prepared by methods known in the art. It is noted that non-pharmaceutical acceptable adducts may also be useful for preparing pharmaceutical acceptable adducts and are therefore within the scope of the present invention.

[0039] The compounds of the invention are also intended to include compounds which differ only in the presence of one or more isotopically enriched atoms, e.g., the replacement of hydrogen by deuterium or tritium, or 11 C. 13 C or 14 Carbon enriched in C or 15 Compounds having the present structures except for the replacement of a carbon with an N-enriched nitrogen are within the scope of this invention.

[0040] The term "treatment" or "treating" refers to the administration of a compound or pharmaceutical composition of the present invention to ameliorate or eliminate a disease or one or more symptoms associated with a disease. The term "prevention" or "preventing" includes reducing the risk of the appearance or development of a disease.

[0041] Unless otherwise specified, "%" refers to weight-%.

[0042] By "±0.3° 2θ" it is meant that the peaks describing the PXRD pattern may vary from the stated value by up to 0.3° 2θ. In some embodiments, the peaks may vary from the stated value by up to 0.2° 2θ, such as by 0.1° 2θ or 0.0° 2θ.

[0043] The present invention has several advantageous features, including the following:

[0044] The crystalline forms of QTX125 and its adducts described herein have various unexpected properties.

[0045] First, it has improved photostability compared to other forms, such as amorphous forms.

[0046] Second, it has improved water insolubility at 37° C. compared to other forms, such as amorphous forms.

[0047] Third, the L-lysine (1:2) adduct described herein is unexpectedly more soluble in saline compared to other forms, such as amorphous forms.

[0048] Compounds of Formula I As noted above, the compound of formula I, also referred to herein as QTX125, is 3-(3-furyl)-N-{4-[(hydroxyamino)carbonyl]benzyl}-5-(4-hydroxyphenyl)-1H-pyrrole-2-carboxamide, having the following formula:

[0049] [ka] .

[0050] Methods for preparing the compound of formula I and evidence of its biological activity for various medical treatment applications are described, for example, in WO2018 / 087082, the contents of which are incorporated herein by reference. The compound of formula I can be used directly in the preparation of the crystalline form of the invention, or an adduct may first be formed (see below) before preparing the crystalline form.

[0051] The crystalline form of the compound of formula I according to the present invention has a characteristic PXRD pattern with peaks at 2θ=20.4°, 21.8°, 22.0°, 22.7° and 23.9° (±0.3° 2θ).

[0052] The peaks of the PXRD pattern can be further described in terms of the relative intensity of the peaks. In the following, the relative intensity is calculated as a percentage of the most intense peak as follows: Relative Intensity (%) = [Peak Intensity / Intensity of the most intense peak] x 100. The values ​​of the peak intensities are provided in counts. In this application, vs = very strong (relative intensity ≥ 70% of the most intense peak), s = strong (45% < relative intensity < 70%), m = medium (20% < relative intensity < 45%), w = weak (5% < relative intensity < 20%) and vw = very weak (relative intensity < 5%) are used.

[0053] In some embodiments, for example when the crystalline form is Form 2, the peaks have the following relative intensity profile: 2θ (±0.3° 2θ) = 20.4° (w); 21.8° (vs); 22.0° (s); 22.7° (w); and 23.9° (m).

[0054] In some embodiments, the characteristic PXRD pattern further has at least one peak selected from 9.0° 2θ, 12.6°, 26.6°, 30.5°, 32.2° (±0.3° 2θ).

[0055] Preferably, the characteristic PXRD pattern has at least two, more preferably at least three, more preferably at least four and most preferably all of the above additional peaks.

[0056] For example, a characteristic PXRD pattern can further have the following combination of peaks: 2θ (±0.3° 2θ)=9.0° and 12.6°, or 9.0° and 26.6°, or 9.0° and 30.5°, or 9.0° and 32.2°, or 12.6° and 26.6°, or 12.6° and 30.5°, or 12.6° and 32.2°, or 26.6° and 30.5°, or 26.6° and 32.2°, or 30.5° and 32.2°; or 2θ (±0.3° 2θ)=9.0°, 12.6°, and 26.6°, or 9.0°, 12.6°, and 30.5°, or 9.0°, 12.6°, and 32.2°, or 9.0°, 26.6°, and 30.5°, or 9.0°, 26.6°, and 32.2°, or 9.0°, 30.5°, and 32.2°, or 12.6°, 26.6°, and 30.5°, or 12.6°, 26.6°, and 32.2°, or 12.6°, 30.5°, and 32.2°, or 26.6°, 30.5°, and 32.2°; or 2θ (±0.3° 2θ)=9.0° and 12.6° and 26.6° and 30.5°, or 9.0° and 12.6° and 26.6° and 32.2°, or 9.0° and 12.6° and 30.5° and 32.2°, or 12.6° and 26.6° and 30.5° and 32.2°; or 2θ(±0.3° 2θ)=9.0° and 12.6° and 26.6° and 30.5° and 32.2°.

[0057] In a preferred embodiment, the characteristic PXRD pattern comprises a peak at 2θ=26.6° (±0.3° 2θ).

[0058] In some embodiments, the peaks have the following relative intensity profile: 2θ (±0.3° 2θ) = 9.0° (m), 12.6° (vw), 26.6° (m), 30.5° (w) and 32.2° (w).

[0059] More preferably, the characteristic PXRD pattern of the crystalline form of the compound of formula I according to the present invention has the peaks mentioned in Table A below:

[0060] [Table A]

[0061] More preferably, the peaks in Table A have the following relative intensity profile: 2θ (±0.3° 2θ)=9.0° (m), 12.6° (vw), 13.0° (w), 14.2° (m), 16.5° (m), 16.9° (m), 20.4° (w), 21.8° (vs), 22.0° (s), 22.7° (w), 23.9° (m), 26.6° (m), 30.5° (w) and 32.2° (w).

[0062] In addition to the peaks described above, a characteristic PXRD pattern of a crystalline form of a compound of formula I according to the present invention may further have one or more, for example two or three or all, of the following peaks, with preferred relative intensity profiles given in brackets: 2θ (±0.3° 2θ)=8.4°(w), 10.3°(w), 15.8°(m), 18.8°(s), 20.9°(w), 21.2°(m), 23.2°(w), 23.4°(w), and / or 29.1°(w).

[0063] In a preferred embodiment, the PXRD pattern is substantially similar or the same as the PXRD pattern shown in Figure 2A or Figure 3A or the top or middle row of Figure 4A, or Figure 5A.

[0064] A crystalline form of the compound of formula I having a PXRD pattern substantially similar or identical to the PXRD pattern shown in Figure 2A or Figure 3A or the top or middle row of Figure 4A, or Figure 5A, may be referred to herein as Form 2. That is, the Form 2 crystalline polymorph of the compound of formula I referred to herein has the above characteristics.

[0065] PXRD patterns may be measured with any suitable diffractometer. As an example, the PXRD patterns of the present application were obtained using a PANalytical X'Pert PRO diffractometer equipped with a PixCEL detector. Suitable diffractometers are typically used in transmission geometry. Suitable diffractometers may, for example, use Cu Kα radiation at 1.54056 Å and operate at 40 kV and 40 mA. The measurement range may be 2-38° 2θ. Analysis may be performed by any suitable means, such as suitable software. Any suitable sample preparation method may be used.

[0066] The crystalline forms of the compound of formula I detailed herein preferably have a purity of at least 97%. More preferably, the crystalline forms of the compound of formula I described herein have a purity of at least 97.5%, more preferably 99% or more, such as 98%, even more preferably 98.5% and most preferably 99.5%. The purity values ​​described herein are measured by high performance liquid chromatography (HPLC). Particularly suitable methods are provided in the examples.

[0067] More preferably, the DSC profile of the crystalline form of the compound of Formula I detailed herein exhibits an exothermic peak between 220-225° C., more preferably between 221 and 223° C. Most preferably, the DSC profile of the crystalline form of the compound of Formula I detailed herein is substantially similar or the same as that shown in FIG. 2B.

[0068] Adducts of compounds of formula I In the present invention regarding adducts, the compound of formula I is adducted with at least one other molecule. Typically, this adduct occurs before crystallization. For example, in the exemplified adduct of QTX125 and L-lysine described herein, L-lysine and QTX125 are mixed before crystallization occurs.

[0069] In some embodiments, the adduct is an adduct with an amino acid, such as a natural amino acid. Preferably, the adduct is an adduct with lysine, most preferably L-lysine.

[0070] Preferably, the adduct is a (1:2) adduct, where for every molecule of the compound of Formula I there are two molecules of the other molecule in the crystalline form.

[0071] In a particularly preferred exemplary embodiment, the adduct is a (1:2) adduct of a compound of formula I with L-lysine, i.e., there are two molecules of L-lysine for every molecule of the compound of formula I. This adduct may be prepared by a method substantially as described herein.

[0072] The crystalline form of the adduct of the compound of formula I according to the present invention has a characteristic PXRD pattern with peaks at 2θ=20.4°, 21.8°, 22.0°, 22.7° and 23.9° (±0.3° 2θ), where “±0.3° 2θ” has the meaning given above in connection with the crystalline form of the compound of formula I.

[0073] In some embodiments, such as when the crystalline form is a crystalline of a 1:2 adduct of the compound of formula I and L-lysine, the peaks have the following relative intensity profile: 2θ (±0.3° 2θ) = 20.4° (m); 21.8° (m); 22.0° (vs); 22.7° (vs); and 23.9° (vs). The definitions of relative intensity are as defined above in connection with the crystalline forms of the compound of formula I.

[0074] In some embodiments, the peaks of the crystalline form of the 1:2 adduct of the compound of formula I and L-lysine are characteristic at 2θ=20.6°, 21.8°, 22.3°, 22.7°, and 23.7° (±0.1° 2θ).

[0075] In some embodiments, the characteristic PXRD pattern further comprises at least one peak selected from 2θ=11.2°, 11.7°, 15.1°, 18.0°, and 26.1° (±0.3° 2θ). Preferably, the characteristic PXRD pattern comprises at least two, more preferably at least three, more preferably at least four, and most preferably all of the above additional peaks.

[0076] For example, a characteristic PXRD pattern may further have the following combination of peaks: 2θ (±0.3° 2θ)=11.2° and 11.7°, or 11.2° and 15.1°, or 11.2° and 18.0°, or 11.2° and 26.1°, or 11.7° and 15.1°, or 11.7° and 18.0°, or 11.7 and 26.1°, or 15.1° and 18.0°, or 15.1° and 26.1°, or 18.0° and 26.1°; or 2θ (±0.3° 2θ)=11.2°, 11.7°, and 15.1°, or 11.2°, 11.7°, and 18.0°, or 11.2°, 11.7°, and 26.1°, or 11.7°, 15.1°, and 18.0°, or 11.7°, 15.1°, and 26.1°, or 11.7°, 18.0°, and 26.1°, or 15.1°, 18.0°, and 26.1°; or 2θ (±0.3° 2θ)=11.2° and 11.7° and 15.1° and 18.0°, or 11.2° and 11.7° and 15.1° and 26.1°, or 11.2 and 11.7° and 18.0° and 26.1°, or 11.7° and 15.1° and 18.0° and 26.1°; or 2θ(±0.3° 2θ)=11.2° and 11.7° and 15.1° and 18.0° and 26.1°.

[0077] In some embodiments, the characteristic PXRD pattern comprises a peak at 2θ=26.1° (±0.3° 2θ).

[0078] In some embodiments, the peak has the following relative intensity profile: 2θ (±0.3° 2θ)=11.2°(m), 11.7°(w), 15.1°(w), 18.0°(m), 26.1°(m).

[0079] More preferably, the characteristic PXRD pattern of the crystalline form of the compound of formula I according to the present invention has the peaks mentioned in Table B below:

[0080] [Table B]

[0081] In some embodiments, the peaks in Table B have the following relative intensity profile: 2θ (±0.3° 2θ) = 5.6° (s), 8.2° (vs), 11.2° (m), 11.7° (w), 13.0° (w), 16.9° (w), 17.7° (m), 18.0° (m), 20.4° (m), 21.8° (w), 22.0° (m), 22.7° (s), 23.9° (s), 26.1° (m).

[0082] In addition to the peaks mentioned above, a characteristic PXRD pattern of a crystalline form of the adduct of the compound of formula I according to the present invention may further have one or more, for example two or three or all, of the following peaks, with preferred relative intensity profiles given in brackets: 2θ (±0.3° 2θ)=9.7°(w), 10.8°(w), 15.8°(w), 18.8°(s), 23.2°(s), 23.4°(s) and 24.8°(m).

[0083] In a preferred embodiment, the PXRD pattern is substantially similar or the same as the PXRD pattern shown in FIG.

[0084] The crystalline forms of the adducts of the compounds of formula I detailed herein preferably have a purity of at least 97%. More preferably, the crystalline forms of the adducts of the compounds of formula I described herein have a purity of at least 97.5%, more preferably 99% or greater, such as 98%, even more preferably 98.5% and most preferably 99.5%. As discussed elsewhere, methods for measuring purity typically involve HPLC.

[0085] The DSC profile of the crystalline form of the adduct of the compound of formula I detailed herein further preferably exhibits an exotherm peak between 150-160° C., more preferably between 153 and 157° C.; and / or an exotherm peak between 162-170° C., more preferably between 165 and 169° C.; and / or also an exotherm peak between 180-190° C., more preferably between 182 and 186° C. Most preferably, the DSC profile of the crystalline form of the compound of formula I detailed herein is substantially similar or the same as that shown in FIG.

[0086] Pharmaceutical Compositions The pharmaceutical compositions according to the present invention comprise a crystalline form of the compound of formula I according to the present invention or an adduct thereof.

[0087] The pharmaceutical composition according to the present invention comprises at least one crystalline form of the compound of formula I or its adduct as described herein. In some embodiments, two crystalline forms of the compound of formula I or its adduct may be present in the pharmaceutical composition. For example, the pharmaceutical composition according to the present invention may comprise a crystalline form of the compound of formula I in Form 2, as well as a crystalline form of an adduct of the compound of formula I, such as a 1:2 adduct with lysine.

[0088] In some embodiments, the crystalline form of the compound of formula I or its adduct in the pharmaceutical composition is in particulate form.In such a case, the pharmaceutical composition can be described as a suspension or slurry.In other words, the compound of formula I or its adduct is solid when the pharmaceutical composition is used.

[0089] The particles or crystallites constituting such pharmaceutical compositions may have any suitable average particle size, and the present invention is not limited thereby. For example, the average particle size may be at least 10 μm, at least 15 μm, or at least 25 μm. For example, the average particle size may be up to 100 μm, up to 90 μm, or up to 80 μm. A combination of these values ​​may be used to provide an exemplary average particle size range. Other exemplary average particle size ranges may be from 10 to 100 μm, such as from 15 to 80 μm, or from 25 to 60 μm. In such cases, the average particle size refers to the average particle size taken along the longest length of the particles. The sample size taken for the measurement of the average particle size may be any suitable size, such as 5 particles or 10 particles or 30 particles or 50 particles or more. Suitable measurement methods may include, for example, optical microscopy, or scanning electron microscopy, particularly optical microscopy.

[0090] The particles or crystallites may have any suitable shape, and the application is not intended to be limited thereby. Exemplary shapes include spherical, cubic, pyramidal or rod-like.

[0091] Exemplary final concentrations of QTX125 in a pharmaceutical composition according to the invention are at least 8 mg / mL, optionally up to 20 mg / mL, such as 8.5 mg / mL or more, 9 mg / mL or more, and 9.5 mg / mL or more.

[0092] In some embodiments, a crystalline form of the compound of formula I or its adduct of the present invention is used to prepare a liquid pharmaceutical composition. In such cases, the crystalline form of the compound of formula I or its adduct is dissolved (in a suitable medium) to provide the pharmaceutical composition. In such cases, the compound of formula I is not solid at the time of use.

[0093] The pharmaceutical compositions according to the present invention may contain, in addition to the crystalline form of the compound of formula I or its adduct as described herein, one or more other pharma- ceutically acceptable components well known to those skilled in the art, including, but not limited to, pharma- ceutically acceptable carriers, diluents, excipients, adjuvants, buffers, pH adjusting agents, preservatives, antioxidants, bacteriostatic agents, stabilizers, suspending agents, solubilizing agents, surfactants (e.g., wetting agents), colorants, and isotonic solutes (i.e., those that render the formulation isotonic with the blood or other relevant bodily fluids of the intended recipient). Suitable carriers, diluents, excipients, and the like can be found in standard pharmaceutical texts. For example, Handbook of Pharmaceutical Additives , 2nd Edition (eds. M. Ash and I. Ash), 2001 (Synapse Information Resources, Inc., Endicott, New York, USA), Remington's Pharmaceutical Sciences , 18th edition, Mack Publishing Company, Easton, Pa., 1990; Handbook of Pharmaceutical Excipients , 2nd edition, 1994.

[0094] Optionally, the pharmaceutical composition according to the invention further comprises a buffer (i.e. the composition further comprises a buffer salt dissolved therein). Optionally, the buffer may be selected from the group of MES, Bis-Tris, ADA, ACES, PIPES, MOPSO, BES, MOPS, TES, HEPES, DIPSO, MOBS, TAPSO, Tris-HCl, HEPPSO, POPSO, TEA, EPPS, Tricine, Gly-Gly, Bicine, HEPBS, TAPS, AMPD, TABS, AMPSO, CHES, CAPSO, APS, CHAPS, CABS, phosphate and histidine or a combination of the above.

[0095] Without wishing to be bound by theory, it is believed that the use of a buffer may help to stabilize the composition at physiological pH.

[0096] The concentration of the buffer salt in the aqueous pharmaceutical composition may range from 1 mM to 1M, preferably from 1 mM to 100 mM, preferably from 5 mM to 50 mM, preferably from 5 mM to 20 mM.

[0097] The pharmaceutical compositions may also include counterions and salts, such as sodium counterions, chloride ions or NaCl, dissolved in solution.

[0098] The pharmaceutical compositions may contain, in addition to the active ingredient, which is a crystalline form of a compound of Formula I or an adduct thereof as described herein, one or more other active agents, such as, for example, one or more other therapeutic or prophylactic agents.

[0099] In some embodiments, the crystalline form of the compound of formula I according to the present invention or its adduct or the pharmaceutical composition according to the present invention can be used together with at least one other drug to provide a combination therapy, which can be part of the same composition or can be provided as a separate composition and can be administered at the same time or at different times.

[0100] Optionally, the pharmaceutical composition of the invention comprises: -water - optionally a buffer salt or dissolved salt such as NaCl; - a crystalline form of a compound of formula I or an adduct thereof as described herein; and The pH of the pharmaceutical formulation is between pH 7 and pH 8.

[0101] In some embodiments, the crystalline form of the compound of formula I or its adduct described herein can be used to prepare an injectable aqueous pharmaceutical formulation or a dry pharmaceutical formulation obtained by drying such an aqueous pharmaceutical formulation. Such pharmaceutical formulations contain a crystalline form of the compound of formula I or its adduct and a crystalline form of the compound of formula II:

[0102] [ka]

[0103] (In the formula, each R 1 is independently selected from the following groups: -H or

[0104] [ka]

[0105] R 2 does not exist or C 1-4 Alkyl; Q is selected from the following groups: -H, -SO3 - , -OH, -C(O)R 3 or -C(OH)R 3 2; and R 3 are independently -H or C 1-4 The compound may be prepared from a compound of formula (I) selected from the group consisting of alkyl, wherein the molar ratio of the compound of formula I to the compound of formula II is 1:50-1:2; and the pH of the pharmaceutical formulation is between pH7 and pH8.

[0106] In some embodiments, the compound of formula II is: β-cyclodextrin, (C 1-4 alkyl)-β-cyclodextrin, (hydroxy-C 1-4 alkyl)-β-cyclodextrin, and hydroxypropyl β-cyclodextrin or sulfobutyl ether of β-cyclodextrin, such as β-cyclodextrin sulfobutyl ether (SBβCD).

[0107] Those skilled in the art can determine the desired concentration or amount of the active ingredient in such formulations.Exemplary final concentrations of the compound of formula I are at least 8 mg / mL, optionally up to 20 mg / mL, such as 8.5 mg / mL or more, 9 mg / mL or more, and more preferably 9.5 mg / mL or more.Exemplary molar ratios of the compound of formula I to the compound of formula II are 1:40 to 1:2.5, preferably 1:30 to 1:2.5, preferably 1:25 to 1:2.5, preferably 1:20 to 1:2.5, such as 1:15 to 1:2.5, preferably 1:10 to 1:2.5, preferably 1:9 to 1:2.5, preferably 1:8 to 1:2.5, preferably 1:6 to 1:2.5, more preferably 1:4.5 to 1:2.5.

[0108] Optionally, a pharmaceutical composition according to the invention is substantially free of meglumine.

[0109] Preparation method Crystalline forms of the compound of formula I may be prepared by a process comprising the steps of: (i) adding a compound of formula I to water to form a suspension; (ii) heating the suspension; (iii) adding one or more organic solvents prior to cooling; and (iv) isolating the crystalline form of the compound of formula I or an adduct thereof.

[0110] In a preferred embodiment, the one or more organic solvents comprise one or more of a C1-5 alcohol, tetrahydrofuran (THF) and dioxane. More preferably, the one or more organic solvents comprise, and most preferably consist of, one or more of the following: propanol, ethanol, THF and dioxane; and most preferably all of the above. Preferably, the volume ratio of C1-5 alcohol:THF:dioxane is up to 12.5:10:1.5, for example 6:3:0.867.

[0111] Preferably, the heating is carried out at a temperature of 70 to 120°C, for example at a temperature of 90 to 110°C.

[0112] Preferably, step (iii) is carried out with stirring. Stirring may be provided by any suitable means. Preferably, stirring is carried out for several hours between steps (iii) and (iv).

[0113] In some embodiments, after cooling in step (iii), step (iv) involves isolating the solid by filtration, treating with a solvent by displacement, and drying in vacuum with heating. In some embodiments, the solvent comprises ethanol and water. In some embodiments, the step of treating with a solvent comprises treating with ethanol, then water, then ethanol. In some embodiments, ethanol and water are used in an amount between 1-3 vol (e.g., 2 vol), i.e., 1-3 ml per gram of crystalline form. In some embodiments, drying in vacuum with heating involves heating to 100° C., e.g., to 80° C. or to 70° C. In some embodiments, the heating is at least to 30° C., e.g., to 35° C. or to 40° C. Any combination of these endpoints may be used to provide a suitable range. In some embodiments, the heating is between 30-100° C., e.g., between 40-75° C., e.g., between 40-60° C.

[0114] Crystalline forms of the adduct of the compound of formula I may be prepared by a process comprising the steps of: (i) adding an addition compound to ethanol to form a first mixture; (ii) adding a compound of formula I to water and one or more organic solvents to form a second mixture; (iii) combining the first mixture with the second mixture to form a composition; (iv) cooling the composition; and (v) isolating the crystalline form of the adduct of the compound of Formula I.

[0115] In a preferred embodiment, the adduct is an amino acid, preferably an L-amino acid, preferably lysine, most preferably L-lysine. Preferably, the adduct is as discussed above for the crystalline form of the adduct of the compound of formula I.

[0116] If a 1:1 adduct is desired, the compound to be adducted and the compound of formula I should be present in 1:1 equivalents. If a 1:2 adduct is desired, the compound to be adducted and the compound of formula I should be present in 2:1 equivalents, respectively. The corresponding adducts should be provided in the corresponding equivalent amounts.

[0117] In a preferred embodiment, the one or more organic solvents comprise tetrahydrofuran (THF), and more preferably, the organic solvent is tetrahydrofuran.

[0118] In a preferred embodiment, the adding and combining steps (i) to (iii) are carried out at 55-65°C.

[0119] In a preferred embodiment, the cooling step comprises two cooling steps (iii)-a and (iii)-b. In the first cooling step (iii)-a, the composition is cooled for a relatively short time, e.g., 0.3-1 hour, to reduce the temperature to around 5-15° C. In the second cooling step (iii)-b, the composition is cooled to room temperature for a relatively long time, e.g., several hours, e.g., 2-24 hours, e.g., 10-20 hours.

[0120] In a preferred embodiment, steps (i) to (iv) are carried out with stirring. Stirring may be performed by any suitable means, such as stirring. The stirring device used is not particularly limited, and suitable stirring devices include a vortex mixer, a magnetic stirrer, a helix mixer, or a paddle stirrer.

[0121] Medical uses, treatment methods In a further aspect, the present invention relates to a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof for use in the manufacture of a medicament.

[0122] The present invention also relates to a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof for use in the manufacture of a medicament for the treatment of cancer.

[0123] Alternatively, the present invention relates to a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof for use in the manufacture of a medicament for the treatment of an autoimmune disease.

[0124] In a further aspect, the present invention relates to a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof for use as a medicament.

[0125] Preferably, the present invention relates to a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof for use in the treatment of cancer.

[0126] Alternatively, the present invention relates to a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof for use in the treatment of an autoimmune disease.

[0127] In a further aspect, the present invention relates to a method of treatment comprising administering a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof to a patient in need of such treatment.

[0128] Preferably, the present invention relates to a method for treating cancer comprising administering a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof to a patient in need of cancer treatment.

[0129] Alternatively, the present invention relates to a method for treating an autoimmune disease comprising administering to a patient in need of such treatment a crystalline form of the compound of formula I or an adduct thereof according to the present invention or a pharmaceutical composition comprising a crystalline form of the compound of formula I or an adduct thereof.

[0130] Preferably, the cancer is selected from breast cancer, chronic myeloid (or myeloid) leukemia (CML), colon cancer, lymphoma (such as non-Hodgkin's lymphoma), fibrosarcoma, gastric cancer, glioblastoma, renal cancer, liver cancer, lung cancer, melanoma, nasopharyngeal cancer, oral cancer, orthotopic multiple myeloma, osteosarcoma, ovarian cancer, pancreatic cancer, and prostate cancer.

[0131] Preferably, the autoimmune disease is selected from autoimmune hepatitis; inflammatory demyelinating diseases of the central nervous system; systemic lupus erythematosus; acute anterior uveitis; Sjogren's syndrome; rheumatoid arthritis; type 1 diabetes; Graves' disease; and inflammatory bowel disease.

[0132] Inflammatory demyelinating diseases of the central nervous system are diseases in which the myelin supporting cells and / or myelin layer of the central nervous system, such as oligodendrocytes, are destroyed. Demyelination causes disruption of nerve signals between the brain and other parts of the body, ultimately resulting in a variety of signs and symptoms, including physical, mental, and sometimes psychiatric problems.

[0133] Specific and non-limiting examples of inflammatory demyelinating diseases include multiple sclerosis (MS) (such as relapsing MS, progressive MS, and optic spinal MS); neuromyelitis optica; acute desseminating encephalomyelitis; acute hemorrhagic leukoencephalitis; Baroconcentric sclerosis; Schilder's disease; Marburg MS; oncologic MS; sporadic sclerosis; optic neuritis; transverse myelitis; Susac syndrome; age-related white matter lesions; myalgic encephalomyelitis; Guillain-Barre syndrome; progressive inflammatory neuropathy; leukodystrophies (such as adrenoleukodystrophy and adrenomyeloneuropathy). Preferably, the autoimmune disease is multiple sclerosis or acute desseminating encephalomyelitis. The autoimmune disease is particularly acute desseminating encephalomyelitis, or particularly and most preferably multiple sclerosis.

[0134] Preferably, the autoimmune disease is selected from autoimmune hepatitis and inflammatory demyelinating diseases of the central nervous system.

[0135] In a particularly preferred embodiment, the autoimmune disease is an inflammatory demyelinating disease of the central nervous system, as described above.

[0136] In another particularly preferred embodiment, the autoimmune disease is autoimmune hepatitis.

[0137] The present inventors have found that QTX125, unlike other histone deacetylase inhibitors, advantageously shows no evidence of genotoxicity, particularly clastogenicity or agenicity.Similarly, QTX125 has unexpectedly been observed to have improved pharmacokinetic properties, particularly the best half-life and volume of distribution, compared to other histone deacetylase inhibitors.

[0138] Administration Preferably, the crystalline form of the compound of formula I or its adduct according to the present invention or the pharmaceutical composition comprising the crystalline form of the compound of formula I or its adduct is administered by injection, which may be by infusion (continuous) as well as bolus (discontinuous) administration.

[0139] Methods of administration by injection may be, for example, subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intravesical, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid and intrasternal injection.

[0140] Preferably, the administration is by intravenous infusion or intravenous injection (bolus administration), more preferably by intravenous infusion.

[0141] Subjects / Medications The subject of administration can be any animal. Preferably, the subject is a mammal, such as a rat, mouse, cat, dog, horse, pig, sheep, cow, primate, or human. Preferably, the subject is a human patient.

[0142] Generally, the effective amount of the compound of formula I to be administered will depend on a variety of factors, such as the severity of the disorder being treated and the weight of the subject. The active compound is usually administered one or more times per day, for example, once, twice, three or four times per day, with typical total daily doses ranging from 0.01 to 1,000 mg / kg / day.

[0143] Preferably, the compound of formula I is administered to a human patient at a dose of from 0.5 to 50 mg / kg, preferably from 0.5 to 30 mg / kg, preferably from 1 to 20 mg / kg, more preferably from 5 to 10 mg / kg.

[0144] Preferably, the compound of formula I is administered to a human patient in a dosage of from 25 mg to 4500 mg, preferably from 50 mg to 3000 mg, preferably from 250 mg to 1500 mg per day.

[0145] The compounds of the present invention can be used together with at least one other drug to provide a combination therapy, which can be part of the same composition or provided as separate compositions and can be administered at the same or different times.

[0146] kit Another aspect of the present invention relates to a kit comprising a crystalline form of the compound of formula I or an adduct thereof according to the present invention. The kit also comprises pharma- ceutically acceptable grade water, buffer or saline for use in preparing the dosage form. In some embodiments, the crystalline form of the compound of formula I or an adduct thereof according to the present invention is provided in a separate container from the pharma-ceutically acceptable grade water, buffer or saline in the kit.

[0147] Preferably, the crystalline form of the compound of formula I or its adduct is provided in a suitable container and / or in suitable packaging.

[0148] The kit may also include one or more delivery systems, such as syringes and needles, for delivering or administering the components provided therein, and may also include instructions for use (e.g., instructions for treating a subject).

[0149] Preferably, the kit also includes instructions for use, e.g., written instructions on how to administer the composition (e.g., injection procedures). Most preferably, the kit includes written instructions on how to prepare a suitable pharmaceutical composition from the components provided, and how to subsequently administer the prepared pharmaceutical composition.

[0150] As will be appreciated by those skilled in the art, features and preferred embodiments of one aspect of the invention will also pertain to other aspects of the invention. EXAMPLES

[0151] The following non-limiting examples are provided to illustrate the present invention.

[0152] Example 1 - Characterization of crude QTX125 and Form2 A scale-up process was used to obtain QTX125, and initial characterization and purity assessment of crude QTX125 was performed. Figure 1A shows undissolved substrate decomposing beyond 150°C, which we speculate is likely due to the release of hydroxylamine prior to complete decomposition. Figure 1B shows an overlay of DSC and TGA, which shows no formal endothermic melting. Instead, a large exothermic event coincides with the onset of decomposition as judged by TGA. A slight phase transition or melting was observed at approximately 150°C and again at 180°C. Figure 1C shows the PXRD profile of crude QTX125. The identified peaks are relatively broad, with an amorphous halo effect evident. The purity of crude QTX125 was determined to be 94.30% by high performance liquid chromatography (HPLC). [HPLC method described separately]. As shown in Table 1, four major impurities were identified.

[0153] [Table 1]

[0154] Here, RRT stands for relative retention time. Relative retention time. Relative retention time was calculated using the following formula: RRT = (T 不純物 / T 基準 ) where T = retention time, the base peak was the QTX125 peak.

[0155] The inventors concluded that the RRT of 1.03 corresponds to the carboxamide of QTX125, i.e.

[0156] [ka]

[0157] The RRT of 1.12 corresponds to the carboxylic acid of QTX125, i.e.

[0158] [ka]

[0159] We believe that this will correspond to the above.

[0160] The HPLC purity determination method used throughout these examples employed the following parameters:

[0161] JPEG2025514066000012.jpg137170

[0162] The main impurity was a carboxylic acid moiety, found to be present at >1.5% at RRT 1.12. A series of extractions were attempted to remove this and other impurities. The solvent tetrahydrofuran (THF) was used to aid in dissolution and allow the carboxylic acid to be selectively washed off. Partitioning between THF, water, and saturated sodium bicarbonate solution resulted in a brown solution, which could be phase separated by addition of brine. A single pass using this method improved the purity of QTX125 to >96% as the major impurity was selectively removed into the liquid (Table 2).

[0163] [Table 2]

[0164] As will now be described, flash evaporation was attempted to provide an amorphous phase from a solution of QTX125. Approximately 5.8 g of QXT125 was dissolved in a mixture of THF and water (1:1, 200 mL). To this was added 20 mL of saturated sodium bicarbonate to give a single phase solution. Brine (50 mL) was added, partitioned and the aqueous phase separated. The aqueous phase was back extracted with ethyl acetate (20 ml) and the combined organics concentrated under reduced pressure. The resulting light brown solid was slurried with water (15 ml), filtered and dried under vacuum at 45° C. to give a grey / brown powder (80%, JN572C dried).

[0165] Predominantly amorphous material was successfully isolated.

[0166] Alternatively, crash precipitation was attempted by dissolving crude QTX125 (1 g) in hot DMSO (3 mL) and adding it to ice-cold water (20 mL). After stirring for 10 min, the solid was filtered, returned to the vessel, slurried in water (15 mL), filtered, washed (15 mL), and pulled dry. The resulting grey / brown solid was dried under vacuum at 45 °C (98% yield, 95.77%, (rrt 0.89 / 0.90%, rrt 1.12 / 1.25%)). This yielded a filterable, very poorly crystalline (mostly amorphous) material. This isolation and potential purification method was also observed to increase purity to >95%, but at the expense of a major impurity (identified with an RRT of 1.03, increasing from 0.78% to 0.97%).

[0167] Next, the THF partitioning method was combined with crash precipitation. Approximately 5.8 g of crude QTX125 was dissolved in a mixture of THF (100 mL) and water (100 mL). 20 mL of saturated sodium bicarbonate was added to this mixture to obtain a single-phase solution. Then, brine (50 mL) was added, partitioned, the aqueous phase was separated, and the process was repeated.

[0168] The aqueous phase was then back-extracted with ethyl acetate (20 mL) and the combined organics were concentrated in vacuo. The resulting light brown solid was slurried with water (15 mL), filtered, and dried under vacuum at 45° C. to give a grey / brown powder. A hot solution of crude QTX125 in DMSO (1 g, 3 mL) was then polish-filtered into ice-cold water (20 mL) to induce rapid precipitation. After stirring for 10 min, the solid was filtered, returned to the vessel, slurried with water (15 mL), filtered, washed (15 mL), and pulled dry. The final purity of QTX125 was found to be slightly reduced to above 95.3%. However, more importantly, a new polymorph of QTX125 was identified. This entity, QTX125 “Form 2”, was clearly defined by PXRD, as shown in Figure 2A-C. In all examples, PXRD data were recorded in the range 2-38° 2θ using a PANalytical X'Pert PRO diffractometer equipped with a PixCEL detector used in transmission geometry (X-ray wavelength 1.54056 Å, Cu Kα radiation, 40 kV, 40 mA), with a typical step width of 0.013° 2θ and a measurement time of 25 s per step.

[0169] Briefly, the profile of Form2 observed by DSC is nearly similar to that of crude QTX125, with a minor exotherm preceding the major decomposition event. The decomposition of Form2 begins at a higher temperature than crude QTX125, as shown in the DSC overlay shown in Figure 2C. Note that the minor endothermic transition is also removed. The peak of Form2 is observed at a temperature approximately 10 °C higher than that of crude QTX125 (Figure 2C), suggesting that Form2 is more thermodynamically stable than crude TX125, which is also supported by the PXRD profile. As shown in Figure 2D, the combined DSC and TGA trace of Form2 shows good overlap of the exothermic decomposition events of both traces, indicating the behavior of a new entity.

[0170] Example 2 - Crystallization and intermediate process scale-up of QTX125 Since the purity of QTX125 was improved by ethanol / water slurry and a new solid product was identified (Example 1), crystallization of QTX125 was investigated. Note that "%th" here means % of theoretical yield. "Uncorrected" means that no purity correction is made, since the purity is assumed to be 100% when calculating the yield.

[0171] 250 mg of QTX125 was suspended in 12 volumes of ethanol / water 5% (v / v) at reflux and 8 volumes of THF were dispensed to give a solution containing QTX125 clumps. This was clarified into a crystallization tube, sealed and left for 48 hours, but no solid formation was observed. The solution was stirred and heated to 50°C under a gentle stream of nitrogen to concentrate the solution. Once solids were observed in the suspension, the mixture was cooled to ambient temperature, isolated by filtration and dried under vacuum at 45°C overnight. A total of 185 mg of QTX125 was recovered (74%th., uncorrected). Chemical purity was assessed to be 98.43% by HPLC with 0.44% acid impurity (RRT 1.12). 1 Residual solvent levels were assessed using H NMR assay: purity was determined to be 98%, with residual ethanol at 0.62% and THF at 0.43%.

[0172] The crystalline species of QTX125 was identified as Form 2, as shown in Figures 3A and 3B. Thermal analysis shows the typical exothermic decomposition of the crystalline form, but in this example a small endothermic event at 233 °C is prominent, which may represent an early melting transformation. TGA analysis is consistent with the description of the decomposition (as reported above for Form 2). The sample is relatively free of solvent and water.

[0173] In summary, small-scale crystallization of QTX125 was successfully demonstrated using a mixture of ethanol, THF and water. The improvement in chemical purity was confirmed by HPLC and 1 Although not optimized, the small-scale crystallization process was deemed to be the most suitable method for purifying QTX125 Form2.

[0174] Scale-up (Reaction I) The small scale crystallization method was scaled up to produce material for stability and solubility studies (Examples 3, 4 and 5). A total of 2.5668g of crude QTX125 was used and 10 volumes of THF were required to obtain a hazy brown mixture before clarification. The isolated solid was dried under vacuum at 50°C, and a total of 1.0617g of QTX125 was recovered (41.36%th., uncorrected). Chemical purity was estimated by HPLC to be 98.21% with 0.19% acid impurity (RTT 1.12). 1 Residual solvent levels were assessed using H NMR assay. Purity was determined to be 98% with 0.26% residual ethanol and 0.17% THF. As shown in Figures 4A to 4C, the crystalline species of QTX125 was again identified as Form 2. The DSC thermograph is nearly identical to that of Form 2 isolated from small scale crystallization, with no low temperature events and characterized by a minor exotherm, endotherm and a main exotherm at 235 °C. Additionally, the TGA thermograph shows no weight loss until 180 °C, after which a weight loss of 8.5% is observed coinciding with the main exotherm. The crystallized product is essentially solvent free, as corroborated by NMR.

[0175] In summary, repeated crystallization of QTX125 has successfully produced Form 2 with high chemical purity, excellent thermal properties, and little residual solvent (ethanol content within ICH limits, THF not exceeding 720 ppm). This is the "first scale-up" referred to elsewhere herein.

[0176] Scale-up (Reaction II) Given the low yield of Form2 from the first scale-up reaction, the crystallization protocol was repeated with a change in solvent regime to assess whether recovery of QTX125 could be improved while maintaining high chemical purity, this is the "second scale-up" referred to elsewhere herein.

[0177] A total of 2.571 g of crude QTX125 was suspended in 12.4 volumes of ethanol and 6 volumes of THF and heated to reflux with stirring. 4 mL of deionized water was charged under reflux to give a solution which was clarified in a crystallization flask at 80° C. The solution was stirred and cooled to 50° C. whereupon solid formation was observed. A stream of nitrogen was applied to the solution to concentrate the mixture until solids were observed in suspension. The mixture was returned to 50° C. and slowly cooled to ambient temperature. The solids were isolated by filtration and dried under vacuum at 50° C. to recover a total of 1.7557 g of QTX125 (68.29%th., uncorrected). Chemical purity was estimated to be 97.86% by HPLC with 0.36% acid impurity (RTT 1.12). DMSO 1 Residual solvent levels were assessed using H NMR assay. Purity was determined to be 97% with 0.48% residual ethanol and 0.34% THF. The crystalline species of QTX125 isolated in this manner was confirmed to be Form 2 by PXRD (Figure 5A). As shown in Figure 5B, the DSC and TGA thermographs were nearly identical to those of QTX125 isolated from the first scale-up reaction, characterized by no low-temperature events, a minor exotherm, an endotherm, and a main exotherm at 238 °C, coincident with a weight loss of 8.3%.

[0178] An alternative scale-up procedure is as follows:

[0179] QTX125, 1 wt (g per mL of solvent) was suspended in water (4 vol or 4 mL per g of QTX125) at 100° C. Propanol, 5 vol (i.e. 5 mL per g of QTX125), ethanol, 1 vol (i.e. 1 mL per g of QTX125), THF, 3 vol (i.e. 3 mL per g of QTX125), and dioxane, 0.867 vol (i.e. 0.867 mL per g of QTX125) were added. The solution was clarified into a crystallization vessel at 100° C. and cooled with stirring, during which a solid was observed to form. The mixture was stirred overnight.

[0180] The solids were isolated by filtration and the filter cake was treated with the following solvent: Ethanol, 2 vol (i.e. 2 mL per gram of QTX125) Water, 2 vol (i.e. 2 mL per 1 g of QTX125) Ethanol, 2 vol (i.e. 2 mL per gram of QTX125).

[0181] The solid was dried in vacuum at 50° C. overnight. Recovery: 0.5916 g, 55% th.uncorr. 1H NMR, DMSO, consistent with structure. Residual solvents: dioxane, 0.21%, propanol / ethanol, 0.14%. CP by HPLC. Recovery: 9.26 g, 62.06% th.uncorr. 1H NMR, DMSO, consistent with structure. Residual solvent: present but not quantifiable. CP by HPLC, 99.4 area %. No impurities greater than 0.5 area %. PXRD pattern consistent with Form 2. DSC thermograph, single exotherm at 242 °C consistent with Form 2.

[0182] Example 3 - Crystalline Form of Adduct of Compound of Formula I L-Lysine (2 equiv, 2M) was clarified in similarly clarified ethanol (43 ml, 48 vol (i.e. 48 mL per g QTX125)) with stirring at 60°C. QXT125 (1 wt (g per mL), 1 equiv, 0.9009 g) was dissolved in THF (3.6 ml, 4 vol (i.e. 4 mL per g QTX125)) and water (0.55 ml, 0.6 vol (i.e. 0.6 mL per g QTX125)), clarified to L-lysine solution at 60°C and cooled to 50°C for 0.5 h. The mixture was cooled with stirring over 18 h and stirring was continued at ambient temperature for 24 h. The solid was collected by filtration and the filter cake was washed with ethanol (2 x 10 ml) and dried under vacuum at 50°C.

[0183] Recovery: 1.4743 g, 96.24% th.uncorr. Chemical purity by HPLC: 96.85 area % (0.35% acid impurity, RRT 1.12). 1 H NMR assay: 96%, including residual ethanol (3.02%), with a 1:2 stoichiometry of QTX125 to L-lysine.

[0184] PXRD (Figure 9), TGA and DSC (Figure 10) indicate high crystallinity.

[0185] Example 4 - Evaluation of photostability of QTX125 The photostability of the novel crystalline form of QTX125 was evaluated in the solid state and in solution. Using "forced lighting" conditions, samples were exposed to 12 Klux / hr and 2.8 UV W / m 2 / h. The samples were stored at a temperature of 30° C. After incubation, the chemical purity of QTX125 was measured by HPLC.

[0186] The following indicators are provided to aid in navigation of the data presented in Tables 3.1.1 to 3.3.6: 3.1. Evaluation of photostability of x-QTX125 Form2. 3.2.x-Amorphous QTX125 photostability assessment. 3.3. Evaluation of photostability of x-QTX125 1:2 L-lysine adduct. 3.x.1-Sample (solid state) stored in an unsealed transparent glass bottle was subjected to forced illumination. 3.x.2-Samples (solid state) sealed under nitrogen in transparent glass bottles and subjected to forced illumination. 3.x.3-Samples (solid state) sealed under nitrogen in amber glass bottles illuminated under ambient laboratory conditions. 3.x.4-Samples (solid state) sealed under nitrogen in amber glass bottles and subjected to forced illumination. 3.x.5-Samples (in solution) sealed under nitrogen in amber glass bottles illuminated under ambient laboratory conditions. 3.x.6-Samples (in solution) sealed under nitrogen in amber glass bottles and subjected to forced illumination.

[0187] Summary data comparing the photostability of the novel crystalline forms of QTX125 after 171 hours of incubation are provided in Tables 3.4.1 through 3.4.4.

[0188] Evaluation of photostability of QTX125 Form2 Table 3.1.1 The photostability of QTX125 Form2 was evaluated in the solid state stored in unsealed clear glass vials. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0189] [Table 3.1.1]

[0190] Form 2 is shown to decompose into two major components over a period of 171 hours. The solid material is observed to change color from off-white to dark green.

[0191] Table 3.1.2 The photostability of QTX125 Form2 was evaluated in the solid state sealed under nitrogen in clear glass vials. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0192] [Table 3.1.2]

[0193] Form 2 is shown to decompose into two major components over a period of 171 hours (RRT 1.04 and 1.11). Data also indicate that secondary decomposition is occurring with new impurities identified at RRT 1.08 and 1.20 entities. The solid material was observed to change color from off-white to dark green.

[0194] Table 3.1.3 Photostability of QTX125 Form2 was evaluated in the solid state sealed under nitrogen in amber glass bottles. Samples were illuminated under ambient laboratory conditions. Values ​​provided are percent (%) of HPLC peak area.

[0195] [Table 3.1.3]

[0196] Form 2 shows little signs of decomposition when stored under these conditions.

[0197] Table 3.1.4 The photostability of QTX125 Form2 was evaluated in the solid state sealed under nitrogen in amber glass bottles. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0198] [Table 3.1.4]

[0199] Form2 is shown to degrade slightly. The major decomposition components identified are the same as those from amorphous QTX125 stored in unsealed clear glass bottles (shown in Table 3.2.1) or in clear glass bottles sealed under nitrogen (shown in Table 3.2.2). These results indicate that decomposition of Form2 may be minimized by limiting exposure to light, for example by storing samples in opaque or amber glass containers.

[0200] Table 3.1.5 Photostability of QTX125 Form2 was evaluated in solutions (THF and water) sealed under nitrogen in amber glass bottles. Samples were illuminated under ambient laboratory conditions. Values ​​provided are percent (%) of HPLC peak area.

[0201] [Table 3.1.5]

[0202] Form 2 has been shown to degrade when stored in solution under ambient laboratory conditions, with the primary decomposition components being the same as those from amorphous QTX125 stored in either unsealed clear glass vials (as shown in Table 3.2.1) or sealed clear glass vials under nitrogen (as shown in Table 3.2.2).

[0203] Table 3.1.6 The photostability of QTX125 Form2 was evaluated in solutions (THF and water) sealed under nitrogen in amber glass bottles. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0204] [Table 3.1.6]

[0205] Form2 is shown to degrade when stored in solution under ambient laboratory conditions. The primary decomposition components are the same as those from amorphous QTX125 stored in unsealed clear glass bottles (shown in Table 3.2.1) or in sealed clear glass bottles under nitrogen (shown in Table 3.2.2). Data also indicate that secondary decomposition of the components identified in RRT1.04 may occur.

[0206] Evaluation of photostability of amorphous QTX125 Table 3.2.1 The photostability of amorphous QTX125 was evaluated in the solid state stored in unsealed clear glass vials. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0207] [Table 3.2.1]

[0208] Amorphous QTX125 decomposes into two major components under forced illumination. The solid material was observed to change color from off-white / beige to brown.

[0209] Table 3.2.2 The photostability of amorphous QTX125 was evaluated in the solid state sealed under nitrogen in clear glass vials. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0210] [Table 3.2.2]

[0211] Amorphous QTX125 decomposes into two major components under forced illumination. The solid material was observed to change color from off-white / beige to brown.

[0212] Table 3.2.3 The photostability of amorphous QTX125 was evaluated in the solid state sealed under nitrogen in amber glass bottles. Samples were illuminated under ambient laboratory conditions. Values ​​provided are percent (%) of HPLC peak area.

[0213] [Table 3.2.3]

[0214] Amorphous QTX125 shows slight signs of decomposition when illuminated under ambient laboratory conditions.

[0215] Table 3.2.4 The photostability of amorphous QTX125 was evaluated in the solid state sealed under nitrogen in amber glass bottles. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0216] [Table 3.2.4]

[0217] Amorphous QTX125 decomposes slightly under forced lighting conditions. The major decomposition components are the same as those from amorphous QTX125 stored in unsealed clear glass bottles (shown in Table 3.2.1) or in sealed clear glass bottles under nitrogen (shown in Table 3.2.2). These results indicate that decomposition of amorphous QTX125 may be minimized by limiting exposure to light, for example by storing samples in opaque or amber glass containers.

[0218] Table 3.2.5 The photostability of amorphous QTX125 was evaluated in solutions (THF and water) sealed under nitrogen in amber glass bottles. Samples were illuminated under ambient laboratory conditions. Values ​​provided are percent (%) of HPLC peak area.

[0219] [Table 3.2.5]

[0220] Amorphous QTX125 decomposes when in solution under ambient laboratory conditions. The primary decomposition components are the same as those identified in Tables 3.2.1 and 3.2.2.

[0221] Table 3.2.6 The photostability of amorphous QTX125 was evaluated in solutions (THF and water) sealed under nitrogen in amber glass bottles. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0222] [Table 3.2.6]

[0223] Amorphous QTX125 decomposes when in solution under forced illumination conditions. The major decomposition components are the same as those identified in Tables 3.2.1 and 3.2.2.

[0224] Evaluation of photostability of QTX125 1:2 L-lysine adduct Table 3.3.1 The photostability of QTX125 1:2 L-lysine adduct was evaluated in the solid state stored in unsealed clear glass vials. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0225] [Table 3.3.1]

[0226] The QTX125 1:2 L-lysine adduct decomposes into two major components under forced illumination. No color change in the solid material could be identified.

[0227] Table 3.3.2 The photostability of QTX125 1:2 L-lysine adduct was evaluated in the solid state sealed under nitrogen in clear glass vials. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0228] [Table 3.3.2]

[0229] The QTX125 1:2 L-lysine adduct decomposes into two major components under forced illumination. No color change in the solid material could be identified.

[0230] Table 3.3.3 The photostability of QTX125 1:2 L-lysine adduct was evaluated in the solid state sealed under nitrogen in amber glass bottles. Samples were illuminated under ambient laboratory conditions. Values ​​provided are percent (%) of HPLC peak area.

[0231] [Table 3.3.3]

[0232] QTX125 1:2 L-lysine adduct shows little signs of decomposition when illuminated under ambient laboratory conditions.

[0233] Table 3.3.4 The photostability of QTX125 1:2 L-lysine adduct was evaluated in the solid state sealed under nitrogen in amber glass bottles. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0234] [Table 3.3.4]

[0235] The QTX125 1:2 L-lysine adduct degrades slightly under forced light conditions. The major decomposition components identified are the same as those from L-lysine adducts stored in unsealed clear glass vials (shown in Table 3.3.1) or in clear glass vials sealed under nitrogen (shown in Table 3.3.2). These results indicate that adduct decomposition may be minimized by limiting exposure to light, for example by storing samples in opaque or amber glass containers.

[0236] Table 3.3.5 Photostability of QTX125 1:2 L-lysine adduct was evaluated in solutions (THF and water) sealed under nitrogen in amber glass bottles. Samples were illuminated under ambient laboratory conditions. Values ​​provided are percent (%) of HPLC peak area.

[0237] [Table 3.3.5]

[0238] QTX125 1:2 L-lysine adduct degrades in solution and under lighting under ambient laboratory conditions. The major decomposition components are the same as those identified in Tables 3.3.1 and 3.3.2.

[0239] Table 3.3.6 The photostability of QTX125 1:2 L-lysine adduct was evaluated in solutions (THF and water) sealed under nitrogen in amber glass bottles. Samples were subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0240] [Table 3.3.6]

[0241] QTX125 1:2 L-lysine adduct degrades when in solution and under forced illumination. The major decomposition components are the same as those identified in Tables 3.3.1 and 3.3.2. These data indicate that storing the L-lysine adduct in solution accelerates decomposition and is therefore not recommended.

[0242] Summary Data Summary data comparing the photostability of QTX125 are provided below in Tables 3.4.1 through 3.4.4.

[0243] Table 3.4.1 Summary data showing photostability of QTX125 after 171 hours incubation. All samples were stored in the solid state, sealed under nitrogen in amber glass bottles, and illuminated under ambient laboratory conditions. Values ​​provided are percent (%) of HPLC peak area.

[0244] [Table 3.4.1]

[0245] In summary, all QTX125 instances show little to no signs of decomposition when stored in amber glass bottles and illuminated under ambient laboratory conditions.

[0246] Table 3.4.2 Summary data showing photostability of QTX125 after 171 hours incubation. All samples were stored in the solid state, sealed under nitrogen in amber glass bottles and subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0247] [Table 3.4.2]

[0248] In summary, while all QTX125 entities show little signs of decomposition when stored in amber glass bottles and illuminated under ambient laboratory conditions, the novel crystalline forms of QTX125 (i.e., Form 2 and the 1:2 L-lysine adduct) appear to exhibit improved photostability compared to amorphous QTX125.

[0249] Table 3.4.3 Summary data showing photostability of QTX125 after 171 hours incubation. All samples were stored in solution, sealed under nitrogen in amber glass bottles and illuminated under ambient laboratory conditions. Values ​​provided are percent (%) of HPLC peak area.

[0250] [Table 3.4.3]

[0251] In summary, the L-lysine 1:2 adduct shows improved photostability in solution compared to amorphous QTX125 and Form2.

[0252] Table 3.4.4 Summary data showing photostability of QTX125 after 171 hours incubation. All samples were stored in solution, sealed under nitrogen in amber glass bottles, and subjected to forced illumination. Values ​​provided are percent (%) of HPLC peak area.

[0253] [Table 3.4.4]

[0254] In summary, the L-lysine 1:2 adduct exhibits improved photostability in solution compared to amorphous QTX125 or Form2.

[0255] conclusion These data show that in the solid state, QTX125 Form 2 and the 1:2 L-lysine adduct show improved photostability compared to amorphous QTX125. Furthermore, the 1:2 L-lysine adduct shows improved photostability compared to both QTX125 Form 2 and amorphous QTX125 when stored in solution.

[0256] Example 5 - Evaluation of the aqueous solubility of QTX125 The solubility of the novel crystalline forms of QTX125 was evaluated in the following aqueous solutions: (i) phosphate buffer, pH 3.5, 0.9% w / v sodium chloride; (ii) phosphate buffer, pH 6.5, 0.9% w / v sodium chloride; (iii) acetate buffer, pH 4.5, 0.9% w / v sodium chloride; (iv) citrophosphate buffer, pH 4.5, 0.9% w / v sodium chloride; (v) sodium chloride, 0.9% w / v, and (vi) deionized water. All assays were completed at 37°C.

[0257] Briefly, 30 mg of QTX125 was dispensed into a container and 5 mL of the appropriate aqueous buffer was added. The mixture was vortexed to suspend and warmed to 37°C. At various intervals, 0.4 mL aliquots of the suspension were removed, filtered, and diluted for testing by HPLC. A single HPLC reference sample was used.

[0258] Evaluation of aqueous solubility of QTX125 Form2 Table 4.1 The solubility of QTX125 Form 2 was evaluated in aqueous buffers (i) to (vi). Values ​​provided are the measured concentrations of QTX125 (mg / mL).

[0259] [Table 4.1]

[0260] The data presented in Table 4.1 are visualized in Figures 6A and 6B. In summary, the solubility of Form2 in phosphate buffer pH 3.5 (i) and pH 6.5 (ii), acetate buffer pH 4.5 (iii), citrophosphate buffer pH 4.5 (iv) and sodium chloride 0.9% w / v (v) is very low. The concentration values ​​are 0.15x10 -3 From 1.76x10 -3 mg.ml -1 The solubility profiles follow the same pattern, showing an initial sharp increase in solubility followed by a decline and increase.

[0261] In contrast, the solubility of Form 2 in deionized water (vi) increased rapidly, reaching a maximum at 7 h, and then gradually decreased. PXRD analysis of the solid recovered at 24 h confirmed the identity of Form 2.

[0262] Assessment of the aqueous solubility of QTX125 1:2 L-lysine adduct Table 4.2 The solubility of QTX125 1:2 L-lysine adduct was evaluated in aqueous buffers (i) to (vi). Values ​​provided are the measured concentrations of QTX125 (mg / mL).

[0263] [Table 4.2]

[0264] The data presented in Table 4.3 are visualized in Figures 7A and 7B. In summary, the solubility of the adduct in phosphate buffers pH 3.5 (i) and pH 6.5 (ii), acetate buffer pH 4.5 (iii), and citrophosphate buffer pH 4.5 (iv) is very low. The concentration values ​​are 0.23x10 -3 From 0.39x10 -3 mg.ml -1The solubility is shown to increase or decrease over time as a result of the adduct dissolving and then precipitating from solution. PXRD analysis of the solids collected at 24 hours identified the precipitate as Form 2, suggesting that the 1:2 L-lysine adduct is converted to Form 2 by maturation in aqueous buffer.

[0265] In contrast, the solubility profiles of the adduct in sodium chloride 0.9% w / v (buffer v) and deionized water (vi) were similar, increasing rapidly and then decreasing after 1 h of incubation. The peak solubility values ​​measured at 1 h were 29.81 x 10 -3 mg.ml -1 and 24.5x10 -3 mg.ml -1 It should be noted that the true peaks may exceed these values ​​since no intermediate measurements were taken between 0 and 1 h.

[0266] conclusion Form 2 showed the highest aqueous solubility in deionized water (buffer vi), 0.02782 mg.ml after 7 h of incubation. -1 In contrast, the 1:2 L-lysine adduct was unexpectedly observed to be soluble in both deionized water (buffer vi) and sodium chloride solution (buffer v). The solubility peaks were 0.02450 mg.ml -1 and 0.02981 mg.ml -1 , reached after 1 h of incubation. Figure 8 provides a comparison of the solubility of the two QTX125 entities in deionized water (buffer vi). Both entities of QTX125 appear to be poorly soluble in phosphate buffers pH 3.5 (i) and pH 6.5 (ii), acetate buffer pH 4.5 (iii) and citrophosphate buffer pH 4.5 (iv) under the experimental conditions tested.

Claims

1. Characterized by a powder X-ray diffraction pattern with peaks at 2θ = 20.4°, 21.8°, 22.0°, 22.7°, and 23.9° (±0.3° 2θ), chemical formula I: 【Chemistry 1】 A crystalline form of the compound or an adduct thereof.

2. 2θ: 9.0°, 11.2°, 11.7°, 12.6°, 15.1°, 18.0°, 24.3°, 26.1°, 26.6°, 30.5°, and 32.2° 2θ (±0.3° 2θ), characterized by having an X-ray powder diffraction pattern with one or more additional peaks at: 9.0°, 11.2°, 11.7°, 12.6°, 15.1°, 18.0°, 24.3°, 26.1°, 26.6°, 30.5°, and 32.2° 2θ (±0.3° 2θ).

3. Table A or Table B below:

2. A crystalline form of the compound of formula I or its adduct as claimed in claim 1, characterized by having a powder X-ray diffraction pattern including the peaks shown in:

4. 2. A crystalline form of the compound of formula I or its adduct as claimed in claim 1, having a powder X-ray diffraction pattern substantially similar or identical to the powder X-ray diffraction pattern shown in FIG. 2A or FIG. 3A or FIG. 5A or FIG.

5. 2. The crystalline form of the compound of formula I according to claim 1, which is Form 2.

6. 2. The crystalline form of the compound of formula I or its adduct according to claim 1, wherein the compound of formula I is a crystalline form of an adduct of the compound of formula I with at least one molecule of lysine.

7. 7. The crystalline form of the compound of formula I or its adduct according to claim 6, wherein the compound of formula I is a crystalline form of a lysine (1:2) adduct in which the compound of formula I is added with two molecules of lysine.

8. 7. The crystalline form of the compound of formula I or its adduct according to claim 6, wherein the lysine is L-lysine.

9. 2. A crystalline form of the compound of formula I or its adduct according to claim 1 having a purity of at least 97%.

10. 10. A pharmaceutical composition comprising a crystalline form of the compound of formula I of claim 1 or an adduct thereof and a pharma- ceutically acceptable excipient or carrier.

11. 13. An in vitro complex comprising a crystalline form of the compound of formula I according to claim 1 or an adduct thereof and histone deacetylase 6 (HDAC6).

12. 10. A method for preparing a crystalline form of the compound of formula I or its adduct according to claim 1, comprising the steps of: (i) adding a compound of formula I to water to form a suspension; (ii) heating the suspension; (iii) adding one or more organic solvents prior to cooling; and (iv) isolating the crystalline form of the compound of formula I or an adduct thereof.

13. 10. A method for preparing a crystalline form of an adduct of a compound of formula I according to claim 1, comprising the steps of: (i) adding an addition compound to ethanol to form a first mixture; (ii) adding a compound of Formula I to water and one or more organic solvents to form a second mixture; (iii) combining the first mixture with the second mixture to form a composition; (iv) cooling the composition; and (v) isolating the crystalline form of the adduct of the compound of Formula I.

14. A crystalline form of the compound of formula I or its adduct obtained by the process of claim 12.

15. A crystalline form of an adduct of a compound of formula I obtained by the method of claim 13.

16. 13. A process for preparing a pharmaceutical composition comprising mixing a crystalline form of the compound of formula I or its adduct as claimed in claim 1 with a pharma- ceutically acceptable excipient or carrier.

17. A pharmaceutical composition obtainable by the method of claim 16.

18. A crystalline form of a compound of formula I or an adduct thereof as defined in claim 1, claim 14 or claim 15, or a pharmaceutical composition as defined in claim 10 or claim 17, for use in a method for inhibiting the function of histone deacetylase 6 in a mammalian subject in need of such inhibition.

19. 19. The use of claim 18, wherein the crystalline form or pharmaceutical composition of the compound of formula I or its adduct is used to treat a proliferative disease or an autoimmune disease in a mammalian subject.

20. 20. The crystalline form or pharmaceutical composition of the compound of formula I or its adduct for use according to claim 19, wherein the proliferative disease is cancer.

21. 21. The crystalline form or pharmaceutical composition of the compound of formula I or its adduct for use according to claim 20, wherein the cancer is a solid tumor, optionally a colon tumor, a pancreatic tumor, a liver tumor or an ovarian tumor.

22. A therapeutic agent comprising a crystalline form of the compound of formula I or an adduct thereof as defined in claim 1, claim 14 or claim 15, or a pharmaceutical composition as defined in claim 10 or claim 17.

23. 23. The method of claim 22, wherein the treatment is treating a proliferative or autoimmune disease in a mammalian subject.

24. 23. The method of claim 22, wherein the treatment is treating cancer in a mammalian subject, optionally wherein the cancer is a solid tumor.

25. 20. Use of a crystalline form of a compound of formula I or an adduct thereof as defined in claim 1, claim 14 or claim 15, or a pharmaceutical composition as defined in claim 10 or claim 17 in the manufacture of a medicament.

26. 27. Use of a crystalline form or pharmaceutical composition of a compound of formula I or its adduct as claimed in claim 25 in the manufacture of a medicament for the treatment of a proliferative or autoimmune disease in a mammalian subject.

27. 27. The use of a crystalline form or pharmaceutical composition of the compound of formula I or its adduct as claimed in claim 26, wherein the proliferative disease is cancer, and optionally the cancer is a solid tumor.

28. 13. A kit comprising pharma- ceutical acceptable grade water, buffer or saline for use in preparing a crystalline form or dosage form of the compound of formula I or an adduct thereof according to claim 1.

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