Crystalline forms of triethylenetetramine tetrahydrochloride and their medical uses

Abstract

The present invention describes a new crystalline form of triethylenetetramine tetrachloride that has improved room temperature stability relative to known forms and dichlorides. The new crystalline form exhibits peaks in the XRPD spectrum at 22.9, 25.4, 25.8, 26.6, 34.6, and 35.3±0.1°2θ, and peaks at 943, 1173, 1527, and 1612±5 cm -1 The crystalline form of triethylenetetramine tetrachloride is characterized by a Raman shift of 1000 nm. The crystalline form of triethylenetetramine tetrachloride is useful for treating Wilson's disease. Also provided is a method for preparing the crystalline form of triethylenetetramine tetrachloride, which comprises transferring an anti-solvent into an aqueous solution of the compound. [Selection diagram] None

Description

Technical Field

[0001] Field of the Invention The present invention relates to the crystalline form of triethylenetetramine tetrachloride (TETA·4HCl) and a method for producing the crystalline form. The present invention further relates to the treatment of Wilson's disease using crystalline triethylenetetramine tetrachloride.

Background Art

[0002] Background of the Invention Triethylenetetramine, or 1,2-ethanediamine, N,N'-bis(2-aminoethyl) (TETA) has the structure of:

[0003]

Chemical formula

[0004] and has the following structure. The dichloride salt (TETA·2HCl) is a polyamine chelating agent for copper (II). Its copper chelating properties make it useful in various conditions, particularly in the treatment of Wilson's disease. Wilson's disease is a hereditary disorder caused by mutations in the Wilson's disease protein (ATP7B gene). This condition leads to the accumulation of copper in the body. The copper chelating ability of TETA·2HCl also leads to considerations for the treatment of many conditions such as visceral damage in diabetic patients, Alzheimer's disease, and cancer (Henriet et al, International Journal of Pharmaceutics 511 (2016) 312 - 321).

[0005] However, although TETA.2HCl has been known for many years to be useful in the treatment of Wilson's disease, it has not been a successful treatment. This is at least partly because it has been found difficult to provide TETA.2HCl in a suitable form that has sufficient stability at room temperature. Thus, patients need to store the tablets under low temperature conditions, which is a cumbersome requirement for a treatment that needs to be taken with every meal throughout their lifetime.

[0006] The study also shows that changes in humidity can affect the stability of the salt. The salt is very sensitive to water and exists in different polymorphs depending on the humidity level. As a result of high humidity, the compound becomes unstable. These stability effects lead to challenges in formulating a drug suitable for the treatment of patients and the need to store the material under special conditions such as low temperature. Therefore, there is a need for an improved treatment for Wilson's disease that can be delivered orally and is stable for a long period under ambient conditions.

[0007] EP 1778618 describes synthetic techniques for producing TETA and its salts, including the 2HCl salt and the 4HCl salt. Only the 2HCl salt is said to be useful in the treatment of Wilson's disease.

[0008] WO 2006 / 027705 describes the synthesis of triethylenetetramine including triethylenetetramine dihydrochloride in Form I and Form II. This document does not mention the crystal form of triethylenetetramine tetrahydrochloride. SUMMARY OF THE INVENTION

[0009] Summary of the Invention The inventors have surprisingly found that a new crystalline form of TETA·4HCl has improved handleability and room temperature stability. Therefore, it is more useful for formulation into drugs than the known forms of the dichloride or tetrachloride salts. Previously known techniques for manufacturing TETA·4HCl (such as anti-solvent crystallization processes carried out at room temperature and processes including high temperature drying steps) lead to the crystalline form described herein as Form A. However, the inventors have found that by carefully controlling the manufacturing conditions, in particular the temperature and rate of crystallization, a new crystalline form, identified herein as Form B, can be produced. This new form has good handleability and good stability and shelf-life characteristics and is therefore beneficial for the manufacture of new formulations for treating Wilson's disease, such as tablets.

[0010] Accordingly, the present invention provides the following features: (i) an XRPD pattern having at least two peaks selected from the peaks of 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1° 2θ; and / or (ii) a Raman spectrum having at least two peaks selected from the peaks of Raman shifts of 943, 1173, 1527 and 1612 ± 5 cm -1 of a crystalline form of triethylenetetramine tetrachloride. The present invention also provides a crystalline form of triethylenetetramine tetrachloride having at least one of the above. which contains, in an amount of 10% by weight or less, Form A of triethylenetetramine tetrahydrochloride having an XRPD pattern with peaks at 25.2 and 35.7 ± 0.1° 2θ, and wherein the peaks of the XRPD pattern are those measured using a wavelength of 1.5418 Å is provided.

[0011] Also provided is a pharmaceutical composition comprising the crystalline form described herein together with one or more pharmaceutically acceptable carriers or diluents.

[0012] A method for producing a crystalline form of triethylenetetramine tetrachloride is also provided, which comprises adding an anti-solvent to an aqueous solution of triethylenetetramine tetrachloride and recovering the resulting crystals, wherein the addition of the anti-solvent is carried out at a temperature of about 20 °C or lower. of the present invention is provided.

[0013] The crystalline form of triethylenetetramine tetrachloride, or a pharmaceutical composition containing triethylenetetramine tetrachloride obtained or obtainable by the methods described herein, is also provided.

[0014] Also provided is the crystalline form or pharmaceutical composition described herein for use in the treatment of a human or animal body by therapy, preferably for the prevention or treatment of Wilson's disease.

[0015] Also provided is a method for the prevention or treatment of Wilson's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the crystalline form or pharmaceutical composition described herein.

[0016] Also provided is the use of the crystalline form or pharmaceutical composition described herein in the manufacture of a medicament for the prevention or treatment of Wilson's disease.

[0017] Certain embodiments of the invention are set forth below: 1. The following features: (i) An XRPD pattern having at least two peaks selected from the peaks of 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1° 2θ; and / or (ii) A Raman spectrum having at least two peaks selected from the peaks of Raman shifts of 943, 1173, 1527 and 1612 ± 5 cm -1 of at least one of triethylenetetramine having a crystalline form tetrahydrochloride which is which contains, in an amount of 10% by weight or less, Form A of triethylenetetramine tetrahydrochloride having an XRPD pattern with peaks at 25.2 and 35.7 ± 0.1° 2θ, and a crystalline form wherein the peaks of the XRPD pattern are those measured using a wavelength of 1.5418 Å

[0018] 2. The crystalline form of embodiment 1 having an XRPD pattern having at least two selected from the peaks of 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1° 2θ.

[0019] ​​3. A crystalline form of embodiment 1 or embodiment 2 having an XRPD pattern with at least three peaks selected from 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1° 2θ.

[0020] 4. A crystalline form of any one of embodiments 1 to 3 having an XRPD pattern with peaks at 25.4, 34.6 and 35.3 ± 0.1° 2θ 。

[0021] 5 . Triethylenetetramine having an XRPD pattern with peaks at 25.2 and 35.7 ± 0.1° 2θ tetrahydrochloride in an embodiment containing 5% by weight or less of form A any one of 1 to 4 of the crystalline form.

[0022] 6 . Essentially (i) an XRPD pattern defined by any one of claims 1 to 4; and / or (ii) a Raman spectrum having at least two peaks selected from the peaks of Raman shifts at 943, 1173, 1527 and 1612 ± 5 cm -1 of triethylenetetramine in the form of form B of any one of the above embodiments. tetrahydrochloride of the crystalline form.

[0023] 7 . A pharmaceutical composition comprising a crystalline form of any one of embodiments 1 to 6 and a pharmaceutically acceptable carrier or diluent.

[0024] 8 . A pharmaceutical composition according to embodiment 6 which is a solid oral dosage form comprising a crystalline form of any one of embodiments 1 to 7 and a pharmaceutically acceptable carrier.

[0025] 9 . Triethylenetetramine having an XRPD pattern with peaks at 25.2 and 35.7 ± 0.1° 2θ tetrahydrochloride in form A by 5An embodiment containing 2% by weight or less, more preferably 2% by weight or less 7 or an embodiment 8 of the pharmaceutical composition

[0026] 10 . An embodiment substantially free of Form A of triethylenetetramine tetrachloride having an XRPD pattern with peaks at 25.2 and 35.7±0.1° 2θ 9 of the pharmaceutical composition

[0027] 11 . Adding an antisolvent to an aqueous solution of triethylenetetramine tetrahydrochloride and recovering the resulting crystals, wherein the addition of the antisolvent is carried out at a temperature of about 20°C or lower described in embodiment 1 triethylenetetramine tetrahydrochloride method for producing the crystalline form

[0028] 12 . An embodiment wherein the addition rate of the antisolvent to the solution is 0.5 ml / min or less per gram of TETA.4HCl dissolved in the aqueous solution 11 The method according to

[0029] 13 . (i) adding an antisolvent to an aqueous solution of TETA.4HCl at a temperature T1, for a period of time t1, and / or at an addition rate R1; (ii) optionally, adding seeds of TETA.4HCl; (iii) optionally, stirring the resulting mixture at T1 for an additional period t1a; (iv) optionally, lowering the temperature to temperature T2 and stirring the mixture for an additional period t2; and (v) recovering the resulting crystals comprising, wherein T1 is about 20°C or lower; T2 is at least 5°C lower than T1; t1 is at least 1 hour, R1 is 0.5 ml / min / g or less, t1a is at least 2 hours, and t2 is at least 30 minutes 11 or an embodiment 12 of the method

[0030] 14 . (i) Adding an antisolvent to an aqueous solution of TETA·4HCl at temperature T1 for a period of time t1 and / or at a rate R1; (ii) Optionally, adding seed crystals of TETA·4HCl; (iii) Optionally, further stirring the resulting mixture at T1 for a period of time t1a; (iv) Lowering the temperature to temperature T2 and further stirring the mixture for a period of time t2; and (v) Recovering the resulting crystals comprising, wherein T1 is about 20 °C or lower; T2 is about 10 °C or lower; t1 is at least 1 hour, R1 is 0.5 ml / min / g or lower, t1a is at least 3 hours, and t2 is at least 30 minutes 13 of the method.

[0031] 15 . (i) Adding an antisolvent to an aqueous solution of TETA·4HCl at temperature T1 for a period of time t1 and / or at a rate R1; (ii) Adding seed crystals of TETA·4HCl; (iii) Stirring the resulting mixture at T1 for a further period of time t1a; (iv) Lowering the temperature to temperature T2 and further stirring the mixture for a period of time t2; and (v) Recovering the resulting crystals comprising, wherein T1 is about 15 °C or lower; T2 is about 5 °C or lower; t1 is at least 1 hour, R1 is 0.2 ml / min / g or lower, t1a is at least 4 hours, and t2 is at least 30 minutes 14 of the method.

[0032] 16 . Further comprising drying the recovered crystals at a temperature of less than about 40 °C, preferably less than about 30 °C 11 ~ 15 any one of the methods.

[0033] 17. A mode of manufacturing a pharmaceutical composition by combining the recovered crystals with a pharmaceutically acceptable carrier 11 ~ 16 Any one of the methods.

[0034] 18 . A mode further including compressing a mixture of the crystals and a pharmaceutically acceptable carrier to form tablets, and optionally sugar-coating or film-coating the tablets 17 The method.

[0035] 19 . A mode 11 ~ 18 A crystal form or a pharmaceutical composition obtained by any one of the methods of

[0036] 20 . Any one of the crystal forms or pharmaceutical compositions of the above modes for use in a method of treating the human or animal body by therapy.

[0037] 21 . A crystal form or a pharmaceutical composition for use in the use of mode 20 for the prevention or treatment of Wilson's disease.

Brief Description of the Drawings

[0038]

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Mode for Carrying Out the Invention

[0039] Detailed Description of the Invention Crystalline TETA·4HCl The crystalline form of triethylenetetramine tetrachloride (TETA.4HCl) described in this specification is distinguishable as Form B. This crystalline form can be characterized by one or more of its XRPD spectrum, its Raman spectrum, its melting point, its FTIR spectrum, and its DVS behavior. Details of each of these characteristics of the crystalline form are described below. Typically, the crystalline form of the present invention is characterized by its XRPD spectrum and / or its Raman spectrum, most preferably by its XRPD spectrum. Accordingly, the crystalline form of the present invention typically has the following characteristics: (i) An X-ray powder diffraction (XRPD) pattern having at least two peaks selected from the peaks at 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1° 2θ; and / or (ii) A Raman spectrum having at least two peaks selected from the peaks at Raman shifts of 943, 1173, 1527 and 1612 ± 5 cm -1 and having at least one of the following.

[0040] Typically, the crystalline form of TETA·4HCl of the present invention has an XRPD pattern having at least two peaks selected from the peaks of 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3±0.1° 2θ. The XRPD pattern preferably has at least three peaks, more preferably at least four peaks, selected from the peaks of 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3±0.1° 2θ. More preferably, at least five or all of these peaks are observed in the XRPD pattern. More preferably, the crystalline form of TETA·4HCl of the present invention has an XRPD pattern having at least two peaks, preferably at least three, four, five, or all peaks, selected from the peaks of 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3±0.05° 2θ. It is particularly preferred that the crystalline form of TETA·4HCl has an XRPD pattern having peaks at 25.4, 34.6 and 35.3±0.1° 2θ, more preferably at 25.4, 34.6 and 35.3±0.05° 2θ.

[0041] Typically, the peaks at 25.4 and 35.3±0.1° 2θ, particularly the peak at 25.4±0.1° 2θ, are the strongest. The peak at 25.4±0.1° 2θ is preferably at least 2 times, more preferably at least 3 times the intensity of the next strongest peak. Typically, the peak at 35.3±0.1° 2θ is at least 2 times the intensity of the next strongest peak.

[0042] Typically, the XRPD pattern of Form B of TETA·4HCl is substantially the same as that shown in FIG. 3 and is represented in FIG.

[0043] XRPD data can be obtained using a PANALYTICAL X'PERT PRO MPD diffractometer. The diffraction data are typically obtained with a powder sample using Cu-K radiation having a characteristic wavelength (λ) of 1.5418 Å. αIt is obtained by exposure to X-rays. The X-rays were generated from a Cu anode supplied with a current of 40 kV and 40 mA. Details of the operating conditions for obtaining XRPD data are described in the Examples section of this specification.

[0044] Typically, the crystalline form of TETA·4HCl of the present invention has a Raman spectrum with shifts at two or more of 943, 1173, 1527 and 1612±5 cm -1 -1. The Raman spectrum shows peaks at 943, 1173, 1527 and 1612±5 cm -1 -1, preferably at least two, preferably three, more preferably all four. It is particularly preferred that the crystalline form of TETA·4HCl has a Raman spectrum with shifts at two or more, preferably three, more preferably all four of 943, 1173, 1527 and 1612±2 cm -1 -1. It is particularly preferred that the crystalline form of TETA·4HCl has a Raman spectrum with shifts at 943 and 1173±5 cm -1 , most preferably at 943 and 1173±2 cm -1 -1. Typically, the Raman spectrum is similar to that shown in Fig. 4 (the upper spectrum).

[0045] The Raman spectrum can be obtained, for example, using a Renishaw RA802 Pharmaceutical Analyser. This can be operated at a laser wavelength of 785 nm. Further operating conditions are described in the Examples section of this specification.

[0046] The crystalline form of TETA·4HCl, Form B, is storage-stable. Accordingly, the XRPD pattern and / or Raman spectrum of a sample of the crystalline form of the present invention stored at 20°C for 6 months, preferably 10 months, more preferably 12 months are typically the same as or substantially the same as those of the crystalline form of the present invention described above. At least preferably 90% by weight, more preferably at least 95% by weight, more preferably at least 98% by weight of a sample of the crystalline form of the present invention stored at 20°C for 6 months, preferably 10 months, more preferably 12 months retains the crystalline form, Form B, described herein.

[0047] The crystalline form of TETA·4HCl, Form B, is stable in a humid environment. Accordingly, the XRPD pattern and / or Raman spectrum of a sample of the crystalline form of the present invention stored at 40°C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months are typically the same as or substantially the same as those of the crystalline form of the present invention described above. At least preferably 90% by weight, more preferably at least 95% by weight, more preferably at least 98% by weight of a sample of the crystalline form of the present invention stored at 40°C and 75% humidity for 1 month is the crystalline form, Form B, described herein. At least preferably 90% by weight, more preferably at least 95% by weight, more preferably at least 98% by weight of a sample of the crystalline form of the present invention stored at 40°C and 75% humidity for 4 months, preferably 6 months retains the crystalline form, Form B, described herein.

[0048] Preferably, the storage stability of the crystalline form of the present invention is determined by the XRPD pattern. Therefore, the XRPD pattern of a sample of the crystalline form of the present invention stored at 20°C for 6 months, preferably 10 months, more preferably 12 months is preferably the same as or substantially the same as that of the crystalline form of the present invention described above. At least preferably 90% by weight, more preferably at least 95% by weight, more preferably at least 98% by weight of the sample of the crystalline form of the present invention stored at 20°C for 6 months, preferably 10 months, more preferably 12 months retains an XRPD pattern that is the same as or substantially the same as that of the crystalline form, Form B, described herein. Further, the XRPD pattern of a sample of the crystalline form of the present invention stored at 40°C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months is preferably the same as or substantially the same as that of the crystalline form of the present invention described above. At least preferably 90% by weight, more preferably at least 95% by weight, more preferably at least 98% by weight of the sample of the crystalline form of the present invention stored at 40°C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months retains an XRPD pattern that is the same as or substantially the same as that of the crystalline form, Form B, described herein.

[0049] Instead, the storage stability of the crystalline form of the present invention is determined by Raman spectrum. Therefore, the Raman spectrum of a sample of the crystalline form of the present invention stored at 20 °C for 6 months, preferably 10 months, more preferably 12 months is preferably the same as or substantially the same as the Raman spectrum of the crystalline form of the present invention described above. Preferably at least 90% by weight, more preferably at least 95% by weight, more preferably at least 98% by weight of the sample of the crystalline form of the present invention stored at 20 °C for 6 months, preferably 10 months, more preferably 12 months retains an XRPD pattern that is the same as or substantially the same as that of the crystalline form, Form B, described herein. Further, the Raman spectrum of a sample of the crystalline form of the present invention stored at 40 °C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months is preferably the same as or substantially the same as that of the crystalline form of the present invention described above. Preferably at least 90% by weight, more preferably at least 95% by weight, more preferably at least 98% by weight of the sample of the crystalline form of the present invention stored at 40 °C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months retains a Raman spectrum that is the same as or substantially the same as that of the crystalline form, Form B, described herein.

[0050] A particular advantage of the crystalline form of the present invention relates to its storage stability. In the storage of tablets obtained from TETA·4HCl of Form A, after storage at 40 °C and 75% humidity for 6 months, discolored patches are observed. Tablets obtained from Form A of aged TETA·4HCl are shown in Fig. 7b. This shows the discoloration of the tablets over time. The present invention, and the provision of Form B of TETA·4HCl, particularly substantially pure Form B of TETA·4HCl, aim to address this problem. Tablets obtained from Form B of TETA·4HCl are considered to have a reduced tendency to discolor over time.

[0051] The crystalline form of the present invention typically has two or more, preferably four or more, more preferably five or six or more, and most preferably all of 1475, 1525, 16010, 2380, 2435, 2580, 2830 and 2880 ± 5 cm-1 has an FTIR spectrum with peaks at. The crystalline form of the present invention preferably has peaks at 1525, 2435 and 2675 ± 5 cm -1 and most preferably at 1526, 2436 and 2674 ± 2 cm -1 has an FTIR spectrum with peaks at. The crystalline form of the present invention has a peak at 943 ± 2 cm in the FTIR spectrum -1 and contains 0 wt% or less of the crystalline form having a peak at. The crystalline form 、1 is preferably free of the crystalline form having a peak at 943 ± 2 cm , preferably or substantially free of the crystalline form having a peak at 943 ± 2 cm -1 .

[0052] The FTIR spectrum is typically an FTIR-ATR spectrum and can be obtained using a Nicolet iS5 FT-IR spectrometer in ATR diamond mode. Specific conditions suitable for obtaining the FTIR spectrum are shown in more detail in Example 4.

[0053] The crystalline form of the present invention typically has a melting temperature of about 260 °C, typically about 259 °C, as measured by DSC. The DSC analysis can be performed as described in Example 4. For example, the analysis can be performed using a Toledo DSC3+ device, providing the sample in a 40 μL sealed aluminum pan with holes in the lid under a nitrogen flush of 50 mL / min.

[0054] Analysis of the crystalline form of the present invention by DVS can also be used to distinguish the crystalline form B of the present invention from Form A. The crystalline form of the present invention typically shows a weight increase of 50 - 59%, typically 54 - 57%, at 90% RH or higher. After completion of the sorption and desorption cycles (0% - 95% RH), the weight increase of the sample is typically 10% or less, preferably 5% or less. This is in contrast to Form A of TETA.4HCl, which shows a weight increase of 14 - 15% after sorption / desorption (0 - 95% RH).

[0055] The crystalline form of TETA.4HCl of the present invention is different from Form A of TETA.4HCl 、1It is contained at 0% by weight or less. The preferred crystal form of TETA·4HCl of the present invention substantially does not contain Form A of TETA·4HCl. As used herein, not containing Form A means that the crystal form contains 5% by weight or less of Form A, preferably 2% by weight or less, more preferably 1% by weight or less, most preferably 0.5% by weight or less, or 0.1% by weight or less of Form A.

[0056] Form A of TETA·4HCl is a crystal form obtained under standard crystallization conditions such as those described in Reference Example 3 herein. Form A is characterized by an XRPD pattern having peaks at 25.2 and 35.7±0.1° 2θ, typically 25.2 and 35.7±0.05° 2θ. The XRPD spectrum of Form A preferably also has peaks at 21.8, 26.9 and 28.2±0.1° 2θ, typically 21.8, 26.9 and 28.2±0.05° 2θ . Form Form A also has peaks at 933 and / or 1513±5 cm -1 Typically at 933 and / or 1513±2 cm -1 And can also be characterized by a Raman spectrum having peaks at. In particular, Form A has peaks at 933, 1167, 1513 and 1604±5 cm -1 Typically at 933, 1167, 1513 and 1604±2 cm -1 And is characterized by a Raman spectrum having peaks at. Typically, Form A is characterized by a Raman spectrum as shown in FIG. 4 (Spectrum below).

[0057] The crystal form of the present invention has an XRPD pattern having peaks at 25.2 and 35.7±0.1° 2θ, or having peaks at 21.8, 25.2, 26.9, 28.2 and 35.7±0.1° 2θ, of the crystal form of TETA·4HCl 、1 0% by weight or less contains; Preferably , the crystalline form of the present invention has an XRPD pattern having peaks at 25.2 and 35.7 ± 0.1° 2θ, or has a crystalline form of TETA.4HCl having an XRPD pattern having peaks at 21.8, 25.2, 26.9, 28.2 and 35.7 ± 0.1° 2θ, It is contained at 5% by weight or less, 2% by weight or less, 1% by weight or less, most preferably 0.5% by weight or less or 0.1% by weight or less.

[0058] The crystalline form of the present invention has peaks at 933 and / or 1513 ± 5 cm -1 Typically at 933 and / or 1513 ± 2 cm -1 Or has peaks at 933, 1167, 1513 and 1604 cm -1 ± 5 cm -1 Typically at 933, 1167, 1513 and 1604 ± 2 cm -1 And has a Raman spectrum of the crystalline form of TETA.4HCl having peaks at these positions. The crystalline form of TETA.4HCl contains 、1 0% by weight or less contains; Preferably , the crystalline form of the present invention has peaks at 933 and / or 1513 ± 5 cm -1 Typically at 933 and / or 1513 ± 2 cm -1 has peaks at or has a Raman spectrum having peaks at 933, 1167, 1513 and 1604 cm -1 ± 5 cm -1 Typically at 933, 1167, 1513 and 1604 ± 2 cm -1 has a crystalline form of TETA.4HCl having peaks at 5% by weight or less, 2% by weight or less, 1% by weight or less, and most preferably 0.5% by weight or more or 0.1% by weight or less.

[0059] The crystalline form of TETA.4HCl contains at least 90% by weight of Form B. Preferably, the crystalline form consists essentially of Form B, i.e., it is substantially pure Form B of TETA.4HCl. When the crystalline form consists essentially of Form B, it contains , at least At least 95% by weight of Form B of TETA.4HCl, more preferably at least 98% by weight, more preferably at least 99% by weight, most preferably at least 99.5% by weight or 99.9% by weight of Form B of TETA.4HCl, where Form B of TETA.3HCl is characterized by the XRPD spectrum and / or Raman spectrum described herein, and preferably Form B of TETA.4HCl is characterized by the XRPD spectrum described herein.

[0060] The crystals of TETA.4HCl described herein are typically provided in a dry form. Accordingly, they typically contain less than 1% by weight of water, preferably less than 0.5% by weight of water, more preferably less than 0.1% or 0.05% by weight of water. All residual solvents are preferably less than 0.1% by weight, more preferably less than 0.5% by weight.

[0061] Process for the preparation of crystalline TETA.4HCl TETA.4HCl can be prepared by techniques known in the art. For example, the TETA free base is commercially available and can be converted to crystalline TETA hydrate and isolated by conventional methods. The TETA hydrate can be treated with aqueous HCl to obtain the TETA.4HCl salt. Typically, the TETA.4HCl salt is isolated in a crude form before recrystallizing as the polymorph of Form B.

[0062] Crystalline forms of TETA.4HCl can typically be obtained from aqueous solutions by an antisolvent crystallization process. Such a process involves adding an antisolvent to an aqueous solution of TETA.4HCl and recovering the resulting crystals. When carried out under standard crystallization conditions, for example, by crystallizing at room temperature or above and / or by methods involving drying at elevated temperature, such methods have been found to lead to a single crystalline form of TETA.4HCl identified herein as Form A. Crystals of Form A have also been obtained by varying the solvent system.

[0063] For example, the inventors used the method described in WO 2006 / 027705 to produce TETA.4HCl and found that these methods led to the production of crystals of Form A. The inventors reproduced Example 17 of WO 2006 / 027705 starting from a mixture of triethylenetetramine isomers and using the crystallization conditions described in Example 17 of WO 2006 / 027705. The product was analyzed by XRPD and the results are shown in Figures 12a - 12c. The resulting product contained the characteristic peaks of Form A of TETA.4HCl. However, the specific peaks identified as characteristic of Form B of TETA.4HCl, particularly those around 35° 2θ and 25.4° 2θ, were absent, suggesting that the product produced was Form A of TETA.4HCl, which is identified as being produced by standard room temperature crystallization.

[0064] The inventors have found that by using the same solvent system but varying the crystallization conditions, particularly the time and temperature of the process, crystals of Form B can be obtained.

[0065] At temperatures of about 20 °C or lower, particularly about 15 °C or lower, TETA.4HCl can be produced as Form B. At about 20 °C to 30 °C, the crystal form produced can depend simply on conditions other than the crystallization temperature. Thus, above about 20 °C, typically further conditions need to be controlled to ensure the production of Form B. In particular, the crystal form produced can depend on the rate of crystallization. Thus, slow crystallization favors the formation of Form B, while faster crystallization favors Form A. Even at temperatures of 15 - 20 °C, some crystals of Form A can be produced unless crystallization is carried out slowly. For example, when an antisolvent is added to form crystals, the antisolvent should preferably be added slowly to the solution to ensure that Form B rather than Form A is produced.

[0066] The crystalline form of the present invention is typically produced by crystallization at a temperature of about 20 °C or lower, preferably about 15 °C or lower, more preferably about 10 °C or lower. In one aspect, the preferred temperature for crystallization is 13 °C or lower, more preferably 7 - 13 °C. At a temperature of about 15 °C or lower, particularly 13 °C or lower, Form B is the thermodynamically preferred form, and crystallization will generally result in substantially pure Form B.

[0067] All steps of the crystallization process are preferably carried out at a temperature of less than 30 °C, preferably about 20 °C or lower, preferably about 15 °C or lower, more preferably about 10 °C or lower. If the temperature exceeds about 15 °C, a mixture of Form A and Form B may be produced. If the temperature exceeds about 30 °C, only Form A will result. To ensure that the product produced is substantially pure Form B, the temperature is preferably maintained at about 15 °C or lower at all times during crystallization. At a temperature between about 15 °C and 20 °C, crystals of Form B can be produced by carrying out crystallization at a slow rate of anti-solvent addition. In particular, the addition of seeds of Form B in combination with slow solvent addition promotes the formation of substantially pure Form B. The slow and controlled addition of the anti-solvent ensures that crystallization develops from the seeds and no separate nucleation of Form A occurs.

[0068] Typically, crystallization is carried out by adding the anti-solvent at a rate of 0.5 ml / min or less of the anti-solvent added to the aqueous solution of TETA·4HCl per gram of TETA·4HCl dissolved in the aqueous solution. Thus, the preferred addition rate is 0.5 ml or less of the anti-solvent per gram of TETA·4HCl per minute, i.e., 0.5 ml / min / g or less. The preferred rate of anti-solvent addition is 0.2 ml / min / g or less, more preferably about 0.1 ml / min / g or less. The preferred addition rate is 0.01 - 0.2 ml / min / g, most preferably 0.01 - 0.1 ml / min / g.

[0069] Therefore, to provide substantially pure form B crystals, crystallization is preferably carried out at about 15 °C or lower and preferably at an addition rate of 5 ml / min / g or lower, more preferably 0.2 ml / min / g or lower, for example about 0.1 ml / min / g. Most preferably, crystallization is carried out at 13 °C or lower, for example 7 - 13 °C, and preferably at an addition rate of less than 0.2 ml / min / g, for example about 0.1 ml / min / g or lower.

[0070] Seeds of form B of TETA·4HCl are preferably added. The seeds can be added before, during, or after the anti-solvent addition, typically before or during the anti-solvent addition, most preferably before the anti-solvent addition. When the seeds are added during or after the anti-solvent addition, they are preferably added before crystal formation is observed.

[0071] A preferred method of crystallization uses TETA·4HCl as the starting material, preferably purified TETA·4HCl. The presence of impurities in the starting material can affect the ability to crystallize the desired polymorph. Therefore, TETA·4HCl is preferably in an isolated form, i.e., it is isolated (and typically purified) from any reaction mixture in which it was produced before crystallization to produce form B is initiated. Further, crude TETA·4HCl is preferably recrystallized before the process to produce form B is initiated. This also provides a higher purity starting material and enables the reliable production of form B crystals by following the methods described herein.

[0072] TETA·4HCl is typically dissolved in an aqueous solution before crystallization. The solution typically contains 0.01 - 10 g of TETA·4HCl per 1 ml of water. The solution preferably contains 0.1 - 5 g of TETA·4HCl per 1 ml of water, and most preferably 0.6 - 1.2 g of TETA·4HCl per 1 ml of water. The volume of the antisolvent used for recrystallization is typically 0.5 ml or less per 1 gram of TETA·4HCl dissolved in the aqueous solution. The preferred amount of the antisolvent is 0.2 ml / g TETA·4HCl or less, more preferably about 0.1 ml / g TETA·4HCl or less. The preferred amount of the antisolvent is 0.01 - 0.2 ml / g TETA·4HCl, and most preferably 0.01 - 0.1 ml / g TETA·4HCl.

[0073] A preferred method for the crystallization of Form B is: (i) adding an antisolvent to an aqueous solution of TETA·4HCl, preferably a stirred aqueous solution, at a temperature T1 for a period of time t1 and / or at an addition rate R1; (ii) optionally, adding seed crystals of TETA·4HCl; (iii) optionally, further stirring the resulting mixture at T1 for a period of time t1a; (iv) optionally, lowering the temperature to temperature T2 and further stirring the mixture for a period of time t2; and (v) recovering the resulting crystals comprising.

[0074] T1 is about 20 °C or lower, preferably about 15 °C or lower, more preferably about 10 °C or lower. To produce substantially pure Form B, T1 is preferably about 15 °C or lower, more preferably about 10 °C or lower.

[0075] The antisolvent can be any solvent in which TETA·4HCl is substantially insoluble. Suitable antisolvents include ethanol, methanol, acetonitrile, propan - 2 - ol, acetone, and 1,4 - dioxane, and mixtures thereof. Methanol and ethanol and mixtures thereof, particularly ethanol, are preferred.

[0076] The solution is typically agitated or mixed, typically by stirring, during addition. Further agitation of the mixture, such as stirring, can be applied during steps (iii) and (iv).

[0077] The crystallization process is typically carried out over a long period. Thus, t1 is typically at least 1 hour, preferably at least 1.5 hours. The addition rate R1 of the antisolvent is typically 0.5 ml / min / g or less. The preferred rate R1 is 0.2 ml / min / g or less, more preferably about 0.1 ml / min / g or less, such as 0.01 - 0.2 ml / min / g, most preferably 0.01 - 0.1 ml / min / g.

[0078] When adding seed crystals, these are typically added before, during, or after the implementation of step (i). Preferably, the seed crystals are added before step (i) or during the addition of the antisolvent in step (i). When adding the seed crystals during or after the addition of the antisolvent, the mixture is typically stirred for a further period, t1a, which is preferably at least 2 hours, such as at least 3 hours or 4 hours, such as about 5 hours. Preferably, a further stirring period at a lower temperature is also included before recovering the crystals. This further stirring step is carried out for a period t2 which is preferably at least 30 minutes. The further stirring step is carried out at a low temperature, T2. T2 is typically lower than T1, preferably at least 5 °C lower than T1, more preferably at least 10 °C lower than T1. T2 is typically about 10 °C or lower, preferably about 5 °C or lower, more preferably about 0 °C. It has been found that an increase in the crystallization time or rate and a decrease in the crystallization temperature provide a higher purity form B. The introduction of seed crystals also helps to improve the purity of the crystal form of form B.

[0079] In the above method, typically, T1 is about 20 °C or lower; T2 is at least 5 °C lower than T1; t1 is at least 1 hour, R1 is 0.5 ml / min / g or lower, t1a is at least 2 hours, and t2 is at least 30 minutes. Preferably, T1 is about 20 °C or lower; T2 is about 10 °C or lower; t1 is at least 1 hour, R1 is 0.5 ml / min / g or lower, t1a is at least 3 hours, and t2 is at least 30 minutes. More preferably, T1 is about 15 °C or lower; T2 is about 5 °C or lower; t1 is at least 1 hour, R1 is 0.2 ml / min / g or lower, t1a is at least 4 hours, and t2 is at least 30 minutes. Crystallization is preferably carried out under an inert atmosphere, such as under nitrogen.

[0080] A preferred crystallization method includes at least the above steps (i), (iv) and (v). A more preferred method includes steps (i), (iii), (iv) and (v). The most preferred method includes all of steps (i) to (v). Therefore, a preferred method for producing TETA·4HCl is: (i) Adding an antisolvent to an aqueous solution of TETA·4HCl, typically a stirred aqueous solution, at a temperature T1, for a period of time t1, and / or at a rate R1; (ii) Optionally, adding seed crystals of TETA·4HCl; (iii) Optionally, further stirring the resulting mixture at T1 for a period t1a; (iv) Lowering the temperature to temperature T2 and further stirring the mixture for a period t2; and (v) Recovering the resulting crystals including therein that T1 is about 20 °C or lower; T2 is about 10 °C or lower; t1 is at least 1 hour, R1 is 0.5 ml / min / g or lower, t1a is at least 3 hours, and t2 is at least 30 minutes.

[0081] An even more preferred method suitable for producing substantially pure Form B of TETA·4HCl is: (i) Adding an antisolvent to an aqueous solution of TETA·4HCl, typically a stirred aqueous solution, at a temperature T1, for a period of time t1, and / or at a rate R1; (ii) Adding seed crystals of TETA·4HCl; (iii) Further stirring the resulting mixture at T1 for a period of time t1a; (iv) Lowering the temperature to temperature T2 and further stirring the mixture for a period of time t2; and (v) Recovering the resulting crystals comprising, wherein T1 is about 15 °C or less; T2 is about 5 °C or less; t1 is at least 1 hour, R1 is 0.2 ml / min / g or less, t1a is at least 4 hours, and t2 is at least 30 minutes.

[0082] The crystals can be recovered by any suitable means, provided that the temperature during the recovery step is maintained below about 40 °C, preferably below about 30 °C. It has been found that higher temperature steps carried out before the crystals are completely dry lead only to crystals of Form A. Most preferably, the recovery of the crystals is carried out at below about 25 °C, for example, about 20 °C or less.

[0083] Methods suitable for recovering the crystals include filtration and centrifugation. The resulting crystals are then typically dried at a temperature below about 40 °C, preferably below about 30 °C. The crystals can be washed, for example, with an antisolvent before drying. Suitable antisolvents for washing are those described above, especially methanol or ethanol, most preferably ethanol. Since heating leads to the production of crystals of Form A, drying is typically by vacuum drying. Vacuum drying below about 40 °C is preferred. Preferably.

[0084] The resulting dried product can be further processed, if necessary, for example, by grinding or granulation. Crystal Form B is substantially stable to grinding.

[0085] When relevant, the recovery and further processing steps such as washing, drying, grinding, etc. are typically carried out under an inert atmosphere such as nitrogen.

[0086] Pharmaceutical Composition and Dosage Form The pharmaceutical composition of the present invention comprises the crystalline form B of TETA·4HCl described herein, together with one or more pharmaceutically acceptable carriers or diluents. The pharmaceutical composition can take any suitable form, but is preferably an oral dosage form. For example, the composition can take the form of tablets, capsules, powders, semi-solids, sustained-release formulations, solutions, suspensions or any other suitable composition. Tablets, capsules and powders, especially tablets, are preferred.

[0087] In an alternative embodiment, the composition is administered parenterally, for example subcutaneously or intravenously.

[0088] The pharmaceutical dosage form can be produced by performing further processing steps on the crystals produced as described herein. Thus, the composition, typically an oral dosage form, can be produced by (a) obtaining the form B of TETA·4HCl, for example using the method described above, (b) optionally grinding and / or granulating the obtained crystals, (c) combining the form B of TETA·4HCl with a pharmaceutically acceptable carrier, and (d) optionally mixing the form B of TETA·4HCl and the carrier. Suitable carriers are further described below. When the oral dosage form is a tablet, the process can further include (e) compressing the mixture to form a tablet and optionally sugar-coating or film-coating the tablet. Alternatively, the solid oral dosage form can be a capsule or a powder. In this case, the method of the present invention can further include (e) packaging the obtained mixture, for example in a capsule. Further standard steps, such as grinding, granulating, sugar-coating or film-coating, can be included in the process.

[0089] The pharmaceutical composition typically comprises up to 85% by weight of TETA·4HCl, for example up to 50% by weight of TETA·4HCl. Preferred compositions are sterile and pyrogen-free.

[0090] Pharmaceutically acceptable carriers suitable for the preparation of oral dosage forms include, for example, solubilizing agents such as cyclodextrin or modified cyclodextrin; diluents such as lactose, dextrose, sucrose, cellulose, corn starch or potato starch; lubricants such as silica, talc, stearic acid, magnesium or calcium stearate, and / or polyethylene glycol; binders such as starch, tragacanth gum, gelatin, syrup, acacia, sorbitol, methyl cellulose, carboxymethyl cellulose or polyvinyl pyrrolidone; disaggregating agents such as starch, alginic acid, alginate or sodium starch glycolate; foaming mixtures; dyes; sweeteners; wetting agents such as lecithin, polysorbate, lauryl sulfate; and non-toxic and pharmacologically inert substances commonly used in pharmaceutical formulations. Such pharmaceutical formulations can be manufactured by known methods, for example by means of mixing.

[0091] The compositions of the present invention typically contain a high proportion of Form B of TETA·4HCl and a small amount of Form A. . This The pharmaceutical compositions of the invention contain Form A of TETA·4HCl at 10% by weight or less. Preferably, the compositions of the present invention are substantially free of Form A of triethylenetetramine tetrachloride. As used herein, substantially free of Form A means that the composition contains Form A of TETA·4HCl at 5% by weight or less, preferably 2% by weight or less, more preferably 1% by weight, or 0.5% by weight or less, and most preferably 0.1% by weight or less.

[0092] Medical use A therapeutically effective amount of the compound of the present invention is administered to a subject. It will be understood that the specific dosage level for a particular subject depends on a variety of factors including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination, and the severity of the particular disease being treated. Optimal dosage levels and dosing frequencies are usually determined by clinical trials.

[0093] A typical daily dose is up to 50 mg per kg of body weight, for example 0.001 - 50 mg per kg of body weight, depending on the age, body weight and condition of the subject to be treated, the type and severity of the disease, and the frequency and route of administration. Preferably, the daily dose level is 0.05 mg - 2 g, preferably 0.1 mg - 10 mg. The compounds of the present invention are typically administered to patients in non-toxic amounts.

[0094] The present invention also provides crystalline forms as defined herein or compositions as defined herein for use in a method of treating the human or animal body by therapy. In particular, the crystalline forms and compositions of the present invention are useful in reducing copper levels in a subject and / or reducing the toxic effects of copper retention in a subject. Accordingly, they are useful in the treatment of disorders and diseases associated with elevated copper levels. In particular, they are useful in the prevention and treatment of Wilson's disease.

[0095] TETA itself is a known treatment for Wilson's disease and diseases and disorders associated with elevated copper levels. After administration of the crystalline form of the present invention, the compound dissolves in the in vivo system, and the therapeutic effect of the crystalline form can be expected to be the same as that of the known forms of TETA.

[0096] The subjects addressed by the present invention can be human or animal subjects, particularly human or mammalian, typically human.

Examples

[0097] Examples Reference Example 1: Synthesis of Crude TETA·4HCl As shown in Figures 1a and 1b, crude TETA·4HCl was prepared.

[0098] Example 2: Synthesis of Form B of TETA·4HCl Form B of TETA·4HCl in substantially pure form was prepared according to the procedure shown in Figure 2.

[0099] Reference Example 3: Synthesis of Form A of TETA·4HCl Crude TETA 4HCl was dissolved in two volumes of purified water under nitrogen, and the resulting solution was then purified by filtration. The reactor was heated to 70 °C (jacket reactor temperature), and while the temperature of the reaction mixture was between 55 and 60 °C, 7 volumes of methanol were added at a rate such that the temperature in the mass remained between 55 and 65 °C to recrystallize the product.

[0100] After stirring at 65 °C (jacket reactor temperature) for at least 30 minutes, the reaction mixture was slowly cooled over a period of at least 2 hours 30 minutes while maintaining a cooling rate of about 5 °C per 30 minutes to a temperature between 30 and 35 °C in the mass. The suspension was then stirred for at least 1 hour at a temperature in the mass maintained between 30 and 35 °C.

[0101] The product was filtered through an enamel Nutsch filter having a membrane porosity of 10 μm and washed twice with 1.5 volumes of methanol. After the last wash, forced filtration was carried out to dry the product. The product was dried in a vacuum oven at 60 °C for at least 14 hours.

[0102] Example 4: Analysis of the crystalline form of TETA.4HCl X-ray powder diffraction The above-described example in 2 A few milligrams of the obtained sample were placed between three polymer foils (Kapton® and polypropylene). Kapton® shows a broad peak in the diffractogram with a weak intensity near 2θ = 5.5°.

[0103] The sample was placed in a PANALYTICAL X'PERT PRO MPD diffractometer configured in transmission mode and analyzed using the conditions shown in Table 1 below. The diffraction data was obtained by exposing the powder sample to Cu-K α X-rays having a characteristic wavelength (λ) of 1.5418 Å. The X-rays were generated from a Cu anode supplied with a current of 40 kV and 40 mA. The analysis was carried out between 2° and 50° (unless otherwise specified). The calibration of the diffractometer was confirmed prior to each analysis 。

[0104] Figure 3 shows the XRPD pattern of the crystal (substantially pure Form B) manufactured according to Example 2.

[0105]

Table 1

[0106] Raman spectroscopy The samples of Example 2 and Reference Example 3 were analyzed by Raman spectroscopy. The Renishaw RA802 Pharmaceutical Analyser was used under the following conditions:

[0107]

Table 2

[0108] The spectra are provided in Figures 4 and in 5 Figure in 5 The lower line represents Example 2 (Form B), while the upper line represents Reference Example 3 (Form A). The arrows indicate the shifts specific to Form B. The Raman spectrum of Example 2 shows bands at 943, 1173, 1527 and 1612 cm -1 The Raman spectrum of Reference Example 3 (Form A) shows peaks at 933, 1167, 1513 and 1604 cm -1 .

[0109] FTIR-ATR analysis The infrared spectra were measured with a Nicolet iS5 FT-IR spectrometer equipped with an iS7 ATR module with the parameters set as follows:

[0110]

Table 3

[0111] For Example 2 (Form B), an infrared spectrum was obtained. The spectrum is shown in FIGS. 8a and 8b.

[0112] DSC analysis DSC analysis was performed using a Mettler Toledo DSC3+ (serial number B531255222) in a 40 μl sealed aluminum pan with a hole in the lid before analysis under a nitrogen flush of 50 mL / min.

[0113] [Table 4]

[0114] An endothermic event corresponding to the melting of the sample was observed. The onset temperature and peak temperature are shown in the following table.

[0115] [Table 5]

[0116] Thermogravimetric analysis (TGA) Thermogravimetric analysis was performed using a Pyris 1 TGA analyzer (serial number 537N7052501) in a sealed aluminum pan with a hole opened before analysis under a nitrogen flush of 20 mL / min.

[0117] [Table 6]

[0118] Thermogravimetric analysis showed a weight loss starting at 225°C, which increased after 289.5°C. This is probably due to decomposition. The weight loss was 10.24%: 2.64% between 225.0 and 289.5°C and 7.60% between 289.5 and 299.0°C.

[0119] Dynamic vapor sorption (DVS) analysis The DVS analysis was performed using the SMS DVS Intrinsic analyser (serial number PF 140088) in an open aluminium pan at 25 °C with a nitrogen purge gas flow rate of 100 ml / min. The stability criterion was a weight change of less than 0.002% over a time frame of 5 minutes (minimum 10 minutes, maximum 100 minutes).

[0120]

Table 7

[0121] DVS isotherm plots are provided in Figures 11a (Example 2) and 11b (Reference Example 3). The DVS analysis performed on these two samples shows significant weight gain at high relative humidity values (above 90% RH). - In Example 2, at least +55.8% - In Reference Example 3, at least +60.8%

[0122] The desorption stage shows different behavior for the two solids. Example 2 reaches approximately the reference weight during the second desorption stage. In Reference Example 3, the weight is +14.4% higher than the reference weight at the end of the second desorption stage (and +7.9% higher than the minimum value reached). It is noteworthy that for the latter, the time limit criterion was reached at these steps (thus, the weight of the sample was not stable).

[0123] Example 5: Heckel test The purpose of the Heckel test is to compress the test material under controlled conditions to derive the yield pressure of the bulk material. A material of known weight is compressed in a 10 mm diameter die with a flat-surface finished punch moving at a set speed. The force on the punch is accurately measured at frequent intervals, while the displacement of the punch is used to calculate the volume of the powder. The yield pressure is calculated at low and high punch speeds to evaluate the time-dependent factor for the deformation of the material. Samples produced in Example 2 and Reference Example 3 were subjected to the Heckel test.

[0124] Method Measurement of True Density by Helium Pychnometry Equipment Used: Micromeritics AccuPyc II 1340 Test Parameters: Cup Size 3.5 cm3 Number of Purging 5 Purging Pressure 19.5 psig Number of Runs 10 Execution Filling Pressure 19.5 psig Equilibrium Rate 0.02 psig Execution Accuracy Yes Percentage of Full Scale 0.05% The test was performed repeatedly (assuming that a variation of less than 2% was achieved).

[0125] Compression A pure drug of known weight is compressed to a theoretical zero porosity using a 10 mm diameter flat-faced punch. A compression simulator was used under the following conditions. Tooling: 10 mm round flat finish Profile: V-shaped profile Punch Speed - Low Speed 0.1 mm / sec Punch Speed - High Speed 300 mm / sec Die Lubrication: Yes, by magnesium stearate in acetone Number of Repetitions: 3 Elastic Correction: Yes

[0126] During compression, the position of the punch tip is accurately measured, the force is measured by a load cell, and records of the main compression parameters are made. Temperature and humidity were monitored at intervals during the test.

[0127] The data was analyzed by a Compaction Analysis software program, Heckel equation:

[0128]

Number

[0129] (where D = relative density of the compact P = applied pressure K = slope of the line in the linear region) using which the value of the yield pressure (Py) was generated [Reference: R.W. Heckel. Trans. Metall. Soc. AIME 221 (1961)1001 - 1008].

[0130] Strain rate sensitivity (SRS) In some materials, the deformation characteristics vary with the rate of the applied force. This can be inferred by calculating the strain rate sensitivity. The yield pressure of high - speed compression is given by the following equation:

[0131]

Equation

[0132] using which it is compared with that of low - speed compression [Reference:R.J. Roberts and R.C. Roe, Chem. Eng. Sci. 42(1987) p903].

[0133] Results True density

[0134]

Table 8

[0135]

Table 9

[0136]

Table 10

[0137]

Table 11

[0138]

Table 12

[0139]

Table 13

[0140]

Table 14

[0141] The Heckel test is a measure of the deformation of the formed tablets. The strength of the compact is an indicator of how the residence time affects the adhesion of the compact. The compacts of the reference examples produced at low speed 3 were found to have a medium tensile strength (6 - 11 pounds). At high speed, the strength decreased to about 4 - 6 pounds. On the other hand, the examples 2 showed a strength of 7 - 12 pounds at low speed, but 9 - 11 pounds at high speed, which indicates that the examples 2 have a greater tensile strength of the compressed product with fast compaction fates. Figure 6 shows the strain rate sensitivity of Example 2 and Reference Example 3 at different production speeds.

[0142] Example 6: Aging of Tablets TETA·4HCl obtained according to Reference Example 3 was compressed to form tablets. An image of the tablets is provided in Figure 7a. The tablets were aged for 6 months at 40 °C and 75% humidity. After aging, it was observed that the tablets had many discolored patches. An image of the aged tablets is provided in Figure 7b.

[0143] Tablets were analyzed by Raman spectroscopy under the same conditions as shown in Example 4 above, and the results were compared with the Raman spectra of Forms A and B of TETA·4HCl. Figure 7c shows the area of the tablet indicating the presence of Form A of TETA·4HCl in the dark region. As is clear from the comparison of Figures 7b and c, the area of Form A of TETA·4HCl correlates with the position of the discolored region on the tablet surface.

[0144] Traces of Form B of TETA·4HCl were detected in tablets that may have been formed under compression of TETA·4HCl to form the tablets. The area of Form B of TETA·4HCl does not correlate with the discolored regions observed in the aged tablets.

[0145] Example 7: Crystallization process Crystallization tests for the preparation of Form B were carried out at 20 °C. The starting solution was prepared in a mixture of ethanol / water (25:75), and ethanol was added until a ratio of (75:25) was reached. The first test was carried out by manually adding ethanol. Subsequently, additional tests were carried out using a syringe pump for slow and controlled addition.

[0146] Manual addition (dropwise) In these tests, the addition was carried out manually using a micropipette. A solution of TETA·4HCl in a (25:75) ethanol / water mixture was placed under stirring and thermostatted at 20 °C. The amounts of starting materials of TETA·4HCl are shown in Table 15.

[0147] The antisolvent (ethanol) was added dropwise at regular intervals. Two addition "rates" were tested. After the addition of the antisolvent, the solid phase was sampled and analyzed by X-ray diffraction to determine the nature of the solid phase. The conditions for X-ray diffraction were as shown in Example 4. The last test was carried out at a slower addition rate using seeds of Form B present at the start of the addition (PE1716E007-L-5). This test led to a solid phase that did not have the signal of Form A observed in the diffractogram.

[0148] The results are shown in Table 15, and the diffractograms obtained after analysis of the solid phase in the suspension at the end of ethanol addition are shown in FIGS. 9a, 9b, and 9c. FIGS. 9a - c start with the lowest line: the reference for Form A (the bottom line in FIGS. 9a - c) Reference for Form B PE1716E007 - L - 5 PE1716E007 - L - 4 PE1716E007 - L - 3 PE1716E007 - L - 2 PE1716E007 - L - 1 PE1716E007 - R - 1 (the top line in FIGS. 9a - c) shows the XRPD diffractograms of.

[0149]

Table 15

[0150] Program addition (continuous) In this series of tests, the addition of the antisolvent was driven by a syringe pump. This enabled continuous addition at a very slow rate. The same protocol as above was used: a solution of the starting material close to saturation was prepared with a 25:75 ethanol / water mixture as shown in Table 16. The solution was then saturated with Form B until a solid phase remained in the suspension: this ensured the presence of seeds of Form B at the start of antisolvent addition.

[0151] (for 750 mg of starting material in solution) Two addition rates: 0.05 mL / min and 0.1 mL / min were tested. Ethanol was added at these selected rates until a 75:25 ethanol / water ratio was obtained. Two additional tests were carried out by adding up to 82:18 corresponding to a concentration of approximately 12.5%, and up to a maximum ratio of 87.5 / 12.5 corresponding to a concentration of approximately 9% (starting material / total weight).

[0152] X-ray diffraction analysis was performed using the conditions shown in Example 4. The results are shown in Table 16 below, and the diffractograms obtained after analysis of the solid phase in the final suspension with ethanol addition are shown in FIGS. 10a to 10c. These show diffraction profiles without the signal of Form A observed in two tests at a final ethanol / water ratio of 75:25 and a test at 87.5:12.5. A small shoulder to the left of the peak at 2θ = 25.4° (corresponding to Form A) in the test at 82:18.

[0153] Figs. 10a to c start from the lowest line: Reference of Form A (the lowest line in FIGS. 10a to c) Reference of Form B PE1716E007-L-6 PE1716E007-L-7 PE1716E007-L-8 XRPD diffractograms of PE1716E007-L-9 (the top line in FIGS. 10a to c) are shown.

[0154] [Table 16]

[0155] Example 8: Crystallization process An overview of the synthesis of triethylenetetramine (trientine) tetrahydrochloride (TETA 4HCl) is shown in the following scheme.

[0156] [Chemical formula]

[0157] In Process I, triethylenetetramine (TETA) is converted to the corresponding triethylenetetramine hydrate (TETA hydrate) by stirring in the presence of water and TBME. Isopropanol is added as an antisolvent and, if necessary, the TETA hydrate is seeded. The TETA hydrate is crystallized, filtered, and isolated. Crude triethylenetetramine tetrahydrochloride (crude TETA 4HCl) is obtained by reacting triethylenetetramine hydrate (TETA hydrate) with an aqueous hydrochloric acid solution in ethanol (Process II). Crude triethylenetetramine tetrahydrochloride (crude TETA 4HCl) is recrystallized from a mixture of purified water and ethanol. Crude triethylenetetramine tetrahydrochloride (crude TETA 4HCl) is further purified by recrystallization from a mixture of purified water and ethanol in the presence of seeds of Form B to obtain triethylenetetramine tetrahydrochloride (TETA 4HCl) (Process III).

[0158] The process produces 110 - 130 kg of TETA 4HCl per batch size from 125 kg of TETA. The overall yield of the synthesis is approximately 50% including two recrystallizations of the crude TETA 4HCl.

[0159] The recrystallization for producing the crystals of Form B is carried out as summarized in the flowcharts of Figures 1a, 1b, and 2, with an additional recrystallization of the crude TETA.4HCl being added before the end of Process II (Figure 1b) and before Process III (Figure 2). The process can be described as follows:

[0160] If the equipment is deactivated, all operations are carried out under a nitrogen flow.

[0161] Process I: Production of Triethylenetetramine Hydrate (TETA Hydrate) · Charge triethylenetetramine (nominal amount 125 kg) into the reactor, followed by TBME (185 ± 5 kg). While maintaining the temperature at ≤ 30°C, add water (about 28 kg) with stirring for ≥ 15 minutes. · To promote crystallization if necessary, seed the solution with triethylenetetramine hydrate (about 0.1 kg) while stirring at 25 - 35 °C. · Add isopropanol (64 ± 1 kg) at 25 - 35 °C. · Heat the suspension at 30 - 40 °C for ≥ 15 minutes and then cool it slowly to 15 - 25 °C over ≥ 90 minutes. · Cool the suspension to - 5 - 5 °C and stir for ≥ 30 minutes. · Filter and centrifuge the product. Then collect samples for analysis (GC assay) and measurement of impurities. · If the sample is sufficiently pure, fill the wet TETA hydrate into a dryer and dry it at ≤ 25 °C until the requirements set in the next step are met. · Analyze the purity of the dried product by GC and appearance, analyze the residual water by KF, analyze the identity by FTIR, and analyze the sulfate ash and residual solvents by GC.

[0162] Step II: Production of crude triethylenetetramine tetrahydrochloride (crude TETA·4HCl) · Dissolve triethylenetetramine hydrate (TETA hydrate) in water (85 ± 1 kg) and acidify by adding concentrated hydrochloric acid aqueous solution (200 ± 5 kg) at ≤ 40 °C over ≥ 1 hour. Check the pH value (target pH = 1.0) and add concentrated hydrochloric acid aqueous solution until pH ≤ 1.0 is satisfied. · Cool the reaction mixture to 15 - 25 °C and stir for ≥ 10 minutes. · Treat the solution with ethanol (672 ± 5 kg) added over ≥ 1.5 hours while maintaining the temperature at ≤ 30 °C. · Cool the suspension to - 5 - 5 °C and stir for ≥ 30 minutes. · Filter the product and wash the solid successively with ethanol (1 × 20 kg, then 3 × 25 kg). · Dissolve crude triethylenetetramine tetrahydrochloride (crude TETA·4HCl) in water (340 ± 10 kg). · Treat the solution with ethanol (909 ± 10 kg) added over ≥ 1.5 hours while maintaining the temperature at ≤ 30 °C. · Cool the suspension to - 5 - 5 °C and stir for ≥ 30 minutes. · Filter the product and wash the solid successively with ethanol (1 x 14 kg, then 3 x 15 kg).

[0163] Step III: Production of triethylenetetramine tetrahydrochloride (TETA·4HCl) · Dissolve the crude triethylenetetramine tetrahydrochloride (crude TETA·4HCl) in water (340 ± 10 kg). · Treat the solution with ethanol (909 ± 15 kg) added over ≥ 1.5 h while maintaining the temperature at 7 - 13°C. · During the addition of ethanol, seed the solution with TETA·4HCl (2 wt%). · Stir the suspension for ≥ 5 h, then cool to -5°C and stir for ≥ 30 min. · Filter the product and wash the solid successively with ethanol (1 x 14 kg, then 3 x 15 kg). · Analyze the purity of a sample of the product by GC. · Dry the product at ≤ 40°C and, if the control parameters for drying loss are met, grind the product. · Transfer the ground product under nitrogen to a double food-quality polyethylene bag, then place it in an aluminum bag and seal. Insert the aluminum bag into an HDPE drum.

[0164] Reprocessing The TETA4HCl obtained after recrystallization is tested for impurities by GC. If the impurity level is too high, Step III can be repeated.

Claims

1. A crystalline form of Form B of triethylenetetramine tetrahydrochloride having an XRPD pattern with peaks at 25.4, 34.6 and 35.3±0.1 degrees 2θ, The content of Form B is at least 95% by weight; the content of triethylenetetramine tetrahydrochloride form A having an XRPD pattern with peaks at 25.2 and 35.7±0.1 degrees 2θ is 5% by weight or less; and A crystalline form in which the peaks in the XRPD pattern are measured using a wavelength of 1.5418 Å.

2. 10. A pharmaceutical composition comprising the crystalline form of claim 1 and a pharma- ceutically acceptable carrier or diluent.

3. 3. The pharmaceutical composition of claim 2, which is a solid oral dosage form comprising the crystalline form of claim 1 and a pharma- ceutically acceptable carrier.

4. 4. The pharmaceutical composition of claim 3, containing less than 2% by weight of Form A of triethylenetetramine tetrahydrochloride having an XRPD pattern with peaks at 25.2 and 35.7±0.1 degrees 2θ.

5. 5. The pharmaceutical composition of claim 3 or claim 4, containing less than 1% by weight of Form A of triethylenetetramine tetrahydrochloride having an XRPD pattern with peaks at 25.2 and 35.7±0.1 degrees 2θ.

6. 2. The crystalline form of claim 1 for use in the prevention or treatment of Wilson's disease.

7. A pharmaceutical composition according to any one of claims 2 to 5 for use in the prevention or treatment of Wilson's disease.

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