Crystalline forms of triethylenetetramine tetrahydrochloride and medical uses thereof

Abstract

The present invention describes a novel crystalline form of triethylenetetramine tetrachloride having room temperature stability over known forms and dichloride salts. The novel crystalline forms are characterized by having peaks at 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 + / -0.1 degrees 2 [theta] and Raman shifts 943, 1173, 152 7 and 1612 + / -5 cm <-1 > in the XRPD spectrum. The crystalline form of triethylenetetramine tetrachloride can be used for the treatment of Wilson disease. Also disclosed is a method of preparing a crystalline form of triethylenetetramine tetrachloride comprising adding an anti-solvent to an aqueous solution of a compound.

Description

[0001] This application is a divisional application with an application date of May 3, 2019, application number 201980006492.6, and invention name “Crystal form of triethylenetetramine tetrahydrochloride and its medical use”. Technical Field

[0002] The present invention relates to crystalline forms of triethylenetetramine tetrachloride (TETA.4HCl) and methods for preparing the crystalline forms. The present invention further relates to the use of crystalline triethylenetetramine tetrachloride to treat Wilson's disease. Background Art

[0003] Triethylenetetramine, or 1,2-ethylenediamine, N,N'-bis(2-aminoethyl) (TETA) has the following structure:

[0004]

[0005] The dichloride salt (TETA.2HCl) is a polyamine chelator for copper (II). Its copper chelating properties make it useful for treating various diseases, especially Wilson's disease. Wilson's disease is a genetic disease caused by mutations in the Wilson disease protein (ATP7B gene). This condition causes copper to accumulate in the body. The copper chelating ability of TETA.2HCl has also led to its use in treating a variety of diseases, such as internal organ damage in diabetic patients, Alzheimer's disease, and cancer (Henriet et al., International Journal of Pharmaceutics 511 (2016) 312-321).

[0006] 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 in part because it has proven difficult to provide TETA.2HCl in a suitable form that is sufficiently stable at room temperature. As a result, patients must store the tablets at lower temperature conditions, which is an onerous requirement for a treatment that must be taken with every meal throughout life.

[0007] Studies have also shown that changes in humidity can affect the stability of the salt. The salt is very sensitive to water and exists in different polymorphic forms depending on the humidity level. High humidity causes the compound to be unstable. These stability effects lead to challenges in formulating drugs suitable for patient treatment and require the storage of materials under special conditions, such as lower temperatures. Therefore, there is a need for improved treatments for Wilson's disease that can be administered orally and are stable under long-term environmental conditions.

[0008] 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 for treating Wilson's disease.

[0009] WO 2006 / 027705 describes the synthesis of triethylenetetramine including triethylenetetramine dichloride forms I and II. This document does not mention crystalline forms of triethylenetetramine tetrahydrochloride. Summary of the invention

[0010] The inventors were surprised to find that the new crystalline form of TETA.4HCl has improved handling properties and room temperature stability. Therefore, compared with the dichloride or known forms of tetrachloride salts, it is more suitable for being formulated into a drug. The previously known techniques for producing TETA.4HCl (e.g., anti-solvent crystallization processes carried out at room temperature and processes including high temperature drying steps) result in the crystalline form described herein being Type A. However, the inventors have found that by carefully controlling the manufacturing conditions, particularly the temperature and crystallization rate, a new crystalline form can be produced, referred to herein as Type B. This new form has good handling properties, as well as good stability and shelf life characteristics, and is therefore conducive to the production of new preparations, such as tablets, for the treatment of Wilson's disease.

[0011] Accordingly, the present invention provides a crystalline form of triethylenetetraamine tetrachloride having at least one of the following characteristics:

[0012] (i) an XRPD pattern having at least two peaks selected from peaks at 22.9, 25.4, 25.8, 26.6, 34.6, and 35.3 ± 0.1° 2θ; and / or

[0013] (ii) With the range from 943, 1173, 1527 and 1612 ± 5 cm -1 A Raman spectrum of at least two peaks selected from the peaks at the Raman shift.

[0014] Also provided is a pharmaceutical composition comprising the crystalline form described herein and one or more pharmaceutically acceptable carriers or diluents.

[0015] Also provided is a method for producing a crystalline form of triethylenetetramine tetrachloride, comprising adding an anti-solvent to an aqueous solution of triethylenetetramine tetrachloride and collecting the obtained crystals, wherein the addition of the anti-solvent is performed at a temperature of about 20° C. or below.

[0016] Also provided are crystalline forms of triethylenetetramine tetrachloride obtainable by or obtained by the methods described herein or pharmaceutical compositions comprising triethylenetetramine tetrachloride.

[0017] Also provided is a crystalline form or a pharmaceutical composition as described herein for use in treating the human or animal body by therapy, preferably in the prevention or treatment of Wilson's disease.

[0018] Also provided is a method of preventing or treating Wilson's disease in a subject in need thereof, the method comprising administering to the subject an effective amount of a crystalline form or a pharmaceutical composition described herein.

[0019] Also provided is the use of the crystalline form or pharmaceutical composition described herein in the preparation of a medicament for preventing or treating Wilson's disease.

[0020] Specific aspects of the invention are set out below:

[0021] 1. A crystalline form of triethylenetetramine tetrachloride having at least one of the following characteristics:

[0022] (i) an XRPD pattern having at least two peaks selected from peaks at 22.9, 25.4, 25.8, 26.6, 34.6, and 35.3 ± 0.1° 2θ; and / or

[0023] (ii) With the range from 943, 1173, 1527 and 1612 ± 5 cm -1 A Raman spectrum of at least two peaks selected from the peaks at the Raman shift.

[0024] 2. The crystalline form according to aspect 1, having an XRPD pattern of at least two peaks selected from peaks at 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1° 2θ.

[0025] 3. The crystalline form according to aspect 1 or aspect 2, which has an XRPD pattern of at least two peaks selected from peaks at 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3±0.1°2θ.

[0026] 4. The crystalline form of any one of aspects 1-3, having an XRPD pattern having peaks at 25.4, 34.6, and 35.3 ± 0.1° 2θ.

[0027] 5. The crystalline form of any one of aspects 1-4, comprising no more than 10 wt% of triethylenetetramine tetrachloride Form A, wherein triethylenetetramine tetrachloride Form A has an XRPD pattern having peaks at 25.2 and 35.7±0.1° 2θ.

[0028] 6. The crystalline form according to aspect 5, comprising no more than 5 wt% of triethylenetetramine tetrachloride Form A, wherein triethylenetetramine tetrachloride Form A has an XRPD pattern having peaks at 25.2 and 35.7±0.1° 2θ.

[0029] 7. The crystalline form according to any one of the preceding aspects, consisting essentially of triethylenetetraaminetetrachloride Form B, having:

[0030] (i) an XRPD pattern as defined in any one of Aspects 1 to 4; and / or

[0031] (ii) With the range from 943, 1173, 1527 and 1612 ± 5 cm -1 A Raman spectrum of at least two peaks selected from the peaks at the Raman shift.

[0032] 8. A pharmaceutical composition comprising the crystalline form according to any one of aspects 1 to 7 and a pharmaceutically acceptable carrier or diluent.

[0033] 9. The pharmaceutical composition according to aspect 8, which is a solid oral dosage form comprising the crystalline form according to any one of aspects 1 to 7 and a pharmaceutically acceptable carrier.

[0034] 10. The pharmaceutical composition according to aspect 8 or aspect 9, comprising no more than 10 wt%, preferably no more than 5 wt%, more preferably no more than 2 wt% of triethylenetetramine tetrachloride Type A, wherein the triethylenetetramine tetrachloride Type A has an XRPD pattern having peaks at 25.2 and 35.7±0.1°2θ.

[0035] 11. The pharmaceutical composition of aspect 10, which is substantially free of triethylenetetraminetetrachloride Form A having an XRPD pattern having peaks at 25.2 and 35.7±0.1° 2θ.

[0036] 12. A method for producing a crystalline form of triethylenetetraamine tetrachloride, the method comprising adding an anti-solvent to an aqueous solution of triethylenetetraamine tetrachloride and collecting the obtained crystals, wherein the addition of the anti-solvent is carried out at a temperature of about 20°C or less.

[0037] 13. The method according to aspect 12, wherein the rate of adding the anti-solvent to the solution is no more than 0.5 ml / min for each gram of TETA.4HCl dissolved in the aqueous solution.

[0038] 14. The method according to aspect 12 or aspect 13, comprising:

[0039] (i) adding an antisolvent to a TETA.4HCl aqueous solution at a temperature T1 for a period of time t1 and / or at an addition rate of R1;

[0040] (ii) optionally adding TETA.4HCl seeds;

[0041] (iii) optionally stirring the resulting mixture at T1 for a further period of time t1a;

[0042] (iv) optionally reducing the temperature to temperature T2 and stirring the mixture for a further period of time t2; and

[0043] (v) collecting the resulting crystals;

[0044] Wherein T1 is about 20°C or less; 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.

[0045] 15. The method according to aspect 14, comprising:

[0046] (i) adding an antisolvent to a TETA.4HCl aqueous solution at a temperature T1 for a period of time t1 and / or at an addition rate of R1;

[0047] (ii) optionally adding TETA.4HCl seeds;

[0048] (iii) optionally stirring the resulting mixture at T1 for a further period of time t1a;

[0049] (iv) reducing the temperature to temperature T2 and stirring the mixture for a further period of time t2; and

[0050] (v) collecting the resulting crystals;

[0051] Wherein T1 is about 20°C or less; T2 is about 10°C or less; t1 is at least 1 hour, R1 is 0.5 ml / min / g or less, t1a is at least 3 hours, and t2 is at least 30 minutes.

[0052] 16. The method according to aspect 15, comprising:

[0053] (i) adding an antisolvent to a TETA.4HCl aqueous solution at a temperature T1 for a period of time t1 and / or at an addition rate of R1;

[0054] (ii) adding TETA.4HCl seed crystals;

[0055] (iii) stirring the resulting mixture at T1 for a further period of time t1a;

[0056] (iv) reducing the temperature to temperature T2 and stirring the mixture for a further period of time t2; and

[0057] (v) collecting the resulting crystals;

[0058] Wherein T1 is about 15°C or below; T2 is about 5°C or below; 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.

[0059] 17. The method according to any one of aspects 12-16, further comprising drying the collected crystals at a temperature below about 40°C, preferably below about 30°C.

[0060] 18. The method according to any one of aspects 12 to 17, wherein the collected crystals are combined with a pharmaceutically acceptable carrier to prepare a pharmaceutical composition.

[0061] 19. The method according to aspect 18, wherein the method further comprises compressing the mixture of the crystals and a pharmaceutically acceptable carrier to form a tablet, and optionally coating the tablet with a sugar coating or a film coating.

[0062] 20. A crystalline form or a pharmaceutical composition obtainable by the method of any one of aspects 12-19.

[0063] 21. A crystalline form or pharmaceutical composition according to any preceding aspect for use in a method of treatment of the human or animal body by therapy.

[0064] 22. Use of the crystalline form or pharmaceutical composition according to aspect 21 in preventing or treating Wilson's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0065] Figure 1a and 1b is a flow chart showing a process for producing crude TETA.4HCl.

[0066] Figure 2 is a flow chart showing a method for producing TETA.4HCl Form B in substantially pure form.

[0067] Figure 3 Depicted are X-ray powder diffraction patterns of TETA.4HCl Form A ("curve 1") and a mixture of TETA.4HCl Form A and Form B ("curve 1+2"). Arrows highlight characteristic peaks of TETA.4HCl Form B.

[0068] Figure 4 The X-ray diffraction pattern of TETA.4HCl Form B is depicted.

[0069] Figure 5a Raman spectra of TETA.4HCl Form A (bottom) and Form B (top) are shown.

[0070] Figure 5bThe upper line shows the spectral coverage of Form A and the lower line shows the spectral coverage of Form B. The Form B peak is highlighted.

[0071] Figure 6 The Heckel plots of TETA.4HCl Form A and TETA.4HCl Form B are depicted.

[0072] Figure 7a Tablets of TETA.4HCl formed from TETA.4HCl Form A are shown. Figure 7b Images of the tablets after aging are shown. Figure 7c Raman analysis of an aged tablet is shown, where the dark areas correspond to the location of TETA.4HCl Form A in the tablet.

[0073] Figure 8a and 8b The FTIR-ATR spectrum of TETA.4HCl Form B is shown. Figure 8a Shown 4000-525cm -1 Full spectrum. Figure 8b The fingerprint area 1800-525cm is shown -1 .

[0074] Figure 9a X-ray powder diffraction analysis of samples of Example 7 after manual addition of ethanol at different speeds is shown and compared with reference patterns of Forms A and B. Figure 9b It shows that at 2θ = 20-29° Figure 9a of the amplification. Fig.9c It shows that at 2θ = 34-37° Figure 9a of the amplification.

[0075] Fig.10a X-ray powder diffraction analysis of a sample of Example 7 after programmed addition of ethanol is shown and compared to reference patterns of Forms A and B. Fig.10b It shows that at 2θ = 20-29° Fig.10a of the amplification. Fig.10c It shows that at 2θ = 34-37° Fig.10a of the amplification.

[0076] Fig.11a and 11b The mass changes with respect to relative humidity for the DVS analysis of Example 2 and Reference Example 3 are shown, respectively.

[0077] Figures 12a to 12c An XRPD spectral analysis of a product produced according to a prior art method is shown. DETAILED DESCRIPTION

[0078] Crystallized TETA.4HCl

[0079] The crystalline form of triethylenetetraamine tetrachloride (TETA.4HCl) described herein is referred to as Form B. The crystalline form can be characterized by one or more of its XRPD spectrum, Raman spectrum, melting point, FTIR spectrum, and 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. Therefore, the crystalline form of the present invention generally has at least one of the following characteristics:

[0080] (i) an X-ray powder diffraction (XRPD) pattern having at least two peaks selected from peaks at 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1° 2θ; and / or

[0081] (ii) With the range from 943, 1173, 1527 and 1612 ± 5 cm -1 A Raman spectrum of at least two peaks selected from the peaks at the Raman shift.

[0082] Typically, the crystalline form of TETA.4HCl of the present invention has an 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. Preferably, the XRPD pattern has at least three peaks selected from the peaks at 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1 ° 2θ, more preferably at least four peaks. More preferably, at least five peaks 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 selected from the peaks at 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.05 ° 2θ, preferably at least three peaks, four peaks, five peaks or all of the peaks. 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.

[0083] Typically, the peaks at 25.4 and 35.3 ± 0.1 ° 2θ are the most intense, especially the peak at 25.4 ± 0.1 ° 2θ. Preferably, the intensity of the peak at 25.4 ± 0.1 ° 2θ is at least twice, more preferably at least three times, the intensity of the next strongest peak. Typically, the intensity of the peak at 35.3 ± 0.1 ° 2θ is at least twice the intensity of the next strongest peak.

[0084] In general, the XRPD pattern of TETA.4HCl Form B is similar to Figure 4 The XRPD patterns shown in are substantially similar.

[0085] XRPD data can be obtained using a PANALYTICAL X'PERT PRO MPD diffractometer. Diffraction data are typically obtained by exposing a powder sample to a characteristic wavelength (λ) of Cu-K α The XRPD data were obtained by X-ray radiation. X-rays were generated from a Cu anode powered at 40 kV and 40 mA. Further details of the operating conditions used to obtain the XRPD data are listed in the Examples section of this article.

[0086] Typically, the crystalline form of TETA.4HCl of the present invention has a Raman spectrum with peaks at 943, 1173, 1527 and 1612±5 cm -1 Preferably, the Raman spectrum is at 943, 1173, 1527 and 1612 ± 5 cm -1 The crystalline form of TETA.4HCl preferably has a Raman spectrum with peaks at 943, 1173, 1527 and 1612±2 cm -1 TETA.4HCl has a displacement at two or more, preferably three, and more preferably all four. Particularly preferred TETA.4HCl has a displacement at 943 and 1173 ± 5 cm -1 , most preferably 943 and 1173±2cm -1 The Raman spectrum has a shift at . Usually, the Raman spectrum is similar to Figure 5a Spectra shown in (upper spectrum).

[0087] Raman spectra 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 listed in the Examples section herein.

[0088] TETA.4HCl Type B crystalline form is storage stable. Thus, typically, the XRPD pattern and / or Raman spectrum of a sample of the crystalline form of the present invention that has been stored at 20°C for 6 months, preferably 10 months, more preferably 12 months is identical or substantially identical to the XRPD pattern and / or Raman spectrum of the crystalline form of the present invention described above. Preferably, at least 90 wt%, more preferably at least 95 wt%, more preferably at least 98 wt% of the sample of the crystalline form of the present invention that has been stored at 20°C for 6 months, preferably 10 months, more preferably 12 months maintains the crystalline form (Type B) described herein.

[0089] TETA.4HCl Type B crystalline form is stable in a humid environment. Therefore, typically, the XRPD pattern and / or Raman spectrum of the crystalline form sample of the present invention that has been stored at 40°C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months is the same or substantially the same as the XRPD pattern and / or Raman spectrum of the crystalline form of the present invention described above. Preferably, at least 90 wt%, more preferably at least 95 wt%, more preferably at least 98 wt% of the crystalline form sample of the present invention that has been stored at 40°C and 75% humidity for 1 month retains the crystalline form (Type B) described herein. Preferably, at least 90 wt%, more preferably at least 95 wt%, more preferably at least 98 wt% of the crystalline form sample of the present invention that has been stored at 40°C and 75% humidity for 4 months, preferably 6 months retains the crystalline form (Type B) described herein.

[0090] Preferably, the storage stability of the crystalline form of the present invention is determined by an XRPD pattern. Therefore, preferably, the XRPD pattern of the crystalline form sample of the present invention that has been stored at 20°C for 6 months, preferably 10 months, more preferably 12 months is the same or substantially the same as the XRPD pattern of the crystalline form of the present invention described above. Preferably, at least 90wt%, more preferably at least 95wt%, more preferably at least 98wt% of the crystalline form sample of the present invention that has been stored at 20°C for 6 months, preferably 10 months, more preferably 12 months maintains the same or substantially the same XRPD pattern as the crystalline form (Type B) described herein. In addition, preferably, the XRPD pattern of the crystalline form sample of the present invention that has been stored at 40°C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months is the same or substantially the same as the XRPD pattern of the crystalline form of the present invention described above. Preferably, at least 90 wt %, more preferably at least 95 wt %, more preferably at least 98 wt % of the crystalline form samples of the present invention that have been stored at 40°C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months, retain the same or substantially the same XRPD pattern as the crystalline form (Type B) described herein.

[0091] Alternatively, the storage stability of the crystalline form of the present invention is determined by Raman spectroscopy. Therefore, preferably, the Raman spectrum of the crystalline form sample of the present invention that has been stored at 20°C for 6 months, preferably 10 months, more preferably 12 months is the same or substantially the same as the Raman spectrum of the crystalline form of the present invention described above. Preferably, at least 90wt%, more preferably at least 95wt%, more preferably at least 98wt% of the crystalline form sample of the present invention that has been stored at 20°C for 6 months, preferably 10 months, more preferably 12 months maintains the same or substantially the same XRPD pattern as the crystalline form (Type B) described herein. In addition, preferably, the Raman spectrum of the crystalline form sample of the present invention that has been stored at 40°C and 75% humidity for 1 month, preferably 4 months, more preferably 6 months is the same or substantially the same as the Raman spectrum of the crystalline form of the present invention described above. Preferably at least 90 wt %, more preferably at least 95 wt %, more preferably at least 98 wt % of the crystalline form samples of the present invention that have been stored at 40° C. and 75% humidity for 1 month, preferably 4 months, more preferably 6 months retain the same or substantially the same Raman spectrum as the crystalline form (Type B) described herein.

[0092] A unique advantage of the crystalline form of the present invention relates to its storage stability. After six months of storage at 40°C and 75% humidity, discolouration patches were observed for tablets obtained from TETA.4HCl Form A. Figure 7b Tablets obtained from TETA.4HCl Form A which has been aged are shown. This shows discoloration of the tablets over time. The present invention and the provision of TEA.4HCl Form B, in particular substantially pure TETA.4HCl Form B, are intended to address this problem. Tablets obtained from TETA.4HCl Form B are believed to have a reduced tendency to discolor over time.

[0093] The crystalline form of the present invention generally has an FTIR spectrum having wavelengths at 1475, 1525, 16010, 2380, 2435, 2580, 2830 and 2880 ± 5 cm -1 Preferably, the crystalline form of the present invention has a FTIR spectrum with peaks at 1525, 2435 and 2675±5 cm -1 At, more preferably at 1526, 2436 and 2674 ± 2cm -1 Preferably, the crystalline form of the present invention contains no more than 50 wt%, such as no more than 40 wt%, preferably no more than 20 wt%, more preferably no more than 10 wt% of the peak at 943 ± 2 cm in the FTIR spectrum. -1 Most preferably, the crystalline form is substantially free of peaks at 943 ± 2 cm -1 A crystal form with a peak at .

[0094] FTIR spectra are typically FTIR-ATR spectra, which can be obtained using a Nicolet iS5 FT-IR spectrometer in ATR diamond mode. Specific conditions suitable for obtaining FTIR spectra are further detailed in Example 4.

[0095] The crystalline form of the present invention generally has a melting temperature of about 260° C., typically about 259° C., as measured by DSC. DSC analysis can be performed as described in Example 4. For example, the analysis can be performed using a Toledo DSC3+ apparatus and the sample is provided in a 40 μL sealed aluminum pan, the lid of which is pierced at a rate of 50 mL / min under a nitrogen purge prior to analysis.

[0096] Analysis of the crystalline forms of the present invention by DVS can also be used to distinguish the crystalline forms B and A of the present invention. The crystalline forms of the present invention typically show a weight gain of 50-59%, typically 54-57%, at 90% RH and above. Typically, after completing the adsorption and desorption cycle (0% to 95% RH), the weight gain of the sample does not exceed 10%, preferably does not exceed 5%. This is in contrast to TETA.4HCl Form A, which shows a weight gain of 14-15% after adsorption / desorption (0-95% RH).

[0097] Typically, the crystalline form of TETA.4HCl according to the invention contains no more than 50 wt%, such as no more than 40 wt%, preferably no more than 20 wt%, more preferably no more than 10 wt% of TETA.4HCl Form A. Preferred crystalline forms of TETA.4HCl according to the invention are substantially free of TETA.4HCl Form A. Substantially free of Form A as described herein means that the crystalline form contains no more than 5 wt% of Form A, preferably no more than 2 wt%, more preferably no more than 1 wt%, most preferably no more than 0.5 wt% or no more than 0.1 wt% of Form A.

[0098] TETA.4HCl Form A is a crystalline form obtained under standard crystallization conditions (e.g., those described herein with reference to Example 3). Form A is characterized by an XRPD pattern having peaks at 25.2 and 35.7 ± 0.1 ° 2θ, typically at 25.2 and 35.7 ± 0.05 ° 2θ. Preferably, the XRPD spectrum of Form A also has peaks at 21.8, 26.9, and 28.2 ± 0.1 ° 2θ, typically at 21.8, 26.9, and 28.2 ± 0.05 ° 2θ. In particular, Form A is characterized by Figure 3 The XRPD pattern shown in ("curve 1") is also characterized by having a peak at 933 and / or 1513 ± 5 cm -1 At, usually at 933 and / or 1513 ± 2cm -1In particular, type A is characterized by having peaks at 933, 1167, 1513, and 1604 ± 5 ​​cm -1 At, usually at 933, 1167, 1513 and 1604 ± 2cm -1 Typically, Type A is characterized by Figure 5a Raman spectrum shown (lower spectrum).

[0099] Preferably, the crystalline form according to the present invention contains no more than 50 wt%, such as no more than 40 wt%, preferably no more than 20 wt%, more preferably no more than 10 wt%, more preferably no more than 5 wt%, no more than 2 wt%, no more than 1 wt%, most preferably no more than less than 0.5 wt% or no more than 0.1 wt% of such TETA.4HCl crystalline form: its XRPD pattern has peaks at 25.2 and 35.7±0.1°2θ, or has peaks at 21.8, 25.2, 26.9, 28.2 and 35.7±0.1°2θ.

[0100] Preferably, the crystalline form according to the present invention comprises no more than 50 wt%, such as no more than 40 wt%, preferably no more than 20 wt%, more preferably no more than 10 wt%, more preferably no more than 5 wt%, no more than 2 wt%, no more than 1 wt%, most preferably no more than 0.5 wt% or no more than 0.1 wt% of a crystalline form of TETA.4HCl having a Raman spectrum at 933 and / or 1513 ± 5 cm -1 At, usually at 933 and / or 1513 ± 2cm -1 There are peaks at 933, 1167, 1513 and 1604 cm-1 ± 5 cm -1 At, usually at 933, 1167, 1513 and 1604 ± 2cm -1 There are peaks.

[0101] Preferably, the crystalline form of TETA.4HCl comprises at least 90 wt% of Form B. More preferably, the crystalline form consists essentially of Form B, i.e. it is substantially pure TETA.4HCl Form B. Wherein the crystalline form consists essentially of Form B, it typically comprises at least 95 wt% of TETA.4HCl Form B, more preferably at least 98 wt%, more preferably at least 99 wt%, most preferably at least 99.5 wt% or 99.9 wt% of TETA.4HCl Form B, wherein TETA.3HCl Form B is characterized by an XRPD spectrum and / or a Raman spectrum as described herein, preferably TETA.4HCl Form B is characterized by an XRPD spectrum as described herein.

[0102] The TETA.4HCl crystals described herein are generally provided in a dry form. Therefore, they generally contain less than 1 wt% water, preferably less than 0.5 wt% water, more preferably less than 0.1 wt% or 0.05 wt% water. The total residual solvent is preferably less than 0.1 wt%, more preferably less than 0.5 wt%.

[0103] Production method of crystalline TETA.4HCl

[0104] TETA.4HCl can be prepared by techniques known in the art. For example, TETA free base is commercially available and can be converted to crystalline TETA hydrate and separated by conventional methods. TETA hydrate can be treated with aqueous HCl to obtain TETA.4HCl salt. Typically, TETA.4HCl salt is isolated in crude form as Type B polymorph before recrystallization.

[0105] TETA.4HCl in crystalline form can be obtained by antisolvent crystallization, usually from aqueous solution. The method involves adding an antisolvent to an aqueous solution of TETA.4HCl and collecting the resulting crystals. When carried out under standard crystallization conditions, such as by crystallization at room temperature or higher, and / or by a method including drying at elevated temperature, it has been found that this method can lead to a single crystalline form of TETA.4HCl, referred to herein as Form A. Form A crystals can be obtained even if the solvent system is changed.

[0106] For example, the present inventors have prepared TETA.4HCl using the methods described in WO 2006 / 027705 and found that these methods are capable of producing Type A crystals. The present inventors started with a mixture of isomers of triethylenetetramine and reproduced Example 17 of WO 2006 / 027705 using the crystallization conditions as described in Example 17 of WO 2006 / 027705. The product was analyzed by XRPD and the results are shown in Figure 12a-12c The obtained product contained characteristic peaks of TETA.4HCl Form A. However, certain characteristic peaks known for TETA.4HCl Form B, particularly those near 35° 2θ and the peak near 25.4° 2θ, were absent, indicating that the product prepared was TETA.4HCl Form A, which is known to be prepared by standard room temperature crystallization.

[0107] The present inventors have found that type B crystals can be obtained by using the same solvent system but changing the crystallization conditions, especially the treatment time and temperature.

[0108] At temperatures of about 20°C or less, and especially at temperatures of about 15°C or less, TETA.4HCl can be prepared as Form B. From about 20°C to 30°C, the crystalline form produced may depend on conditions other than just the crystallization temperature. Therefore, it is often necessary to control other conditions above about 20°C to ensure that Form B is produced. In particular, the crystalline form produced may depend on the crystallization rate. Thus, slow crystallization favors the formation of Form B, while faster crystallization favors Form A. Even at a temperature of 15-20°C, some Form A crystals will still be produced unless the crystallization is carried out slowly. For example, in the case of using anti-solvent addition to form crystals, it is preferred that the anti-solvent be added slowly to the solution to ensure that Form B is produced rather than Form A.

[0109] Typically, the crystalline form of the present invention is produced by crystallization at a temperature of about 20° C. or less, preferably about 15° C. or less, more preferably about 10° C. or less. In one embodiment, the preferred temperature for crystallization is 13° C. or less, more preferably 7-13° C. At a temperature of about 15° C. or less, particularly at a temperature of 13° C. or less, Form B is the thermodynamically favored form, and crystallization will generally result in substantially pure Form B.

[0110] Preferably, all steps in the crystallization process are carried out at a temperature below 30°C, preferably about 20°C or lower, preferably about 15°C or lower, and more preferably about 10°C or lower. When the temperature is above about 15°C, a mixture of Form A and Form B can be generated. When the temperature is above about 30°C, only Form A is generated. In order to ensure that the product generated is substantially pure Form B, it is preferred to maintain the temperature at about 15°C or lower at all times during crystallization. At a temperature between about 15°C and 20°C, Form B crystals can be generated by crystallization at a slow anti-solvent addition rate. In particular, the addition of Form B seeds combined with slow solvent addition promotes the formation of substantially pure Form B. The addition of the anti-solvent in a slow and controlled manner ensures that the crystallization progresses from the seeds and that no separate nucleation of Form A occurs.

[0111] Typically, crystallization is performed by adding an antisolvent to the aqueous solution of TETA.4HCl at an antisolvent addition rate of 0.5 mL / min or less per gram of TETA.4HCl dissolved in the aqueous solution. Thus, a preferred addition rate is 0.5 ml of antisolvent per minute per gram of TETA.4HCl, or less, i.e., 0.5 ml / min / g or less. A preferred rate of antisolvent addition is 0.2 ml / min / g or less, more preferably about 0.1 ml / min / g or less. A preferred addition rate is 0.01-0.2 ml / min / g, most preferably 0.01-0.1 ml / min / g.

[0112] Therefore, in order to provide substantially pure B-type crystals, it is preferred to crystallize at a temperature of about 15°C or less and preferably at an addition rate of 5 ml / min / g or less, more preferably 0.2 ml / min / g or less, such as about 0.1 ml / min / g. Most preferably, crystallization is performed at a temperature of 13°C or less (e.g., from 7-13°C) and preferably at an addition rate of less than 0.2 ml / min / g, such as about 0.1 ml / min / g or less.

[0113] Preferably, seeds of TETA.4HCl Form B are added. The seeds may be added before, during or after the antisolvent addition, typically before or during the antisolvent addition, most preferably before the antisolvent addition. If seeds are added during or after the antisolvent addition, they are preferably added before crystal formation is observed.

[0114] Preferred crystallization methods use TETA.4HCl, preferably pure TETA.4HCl, as a starting material. The presence of impurities in the starting material can affect the ability to crystallize the desired polymorph. Therefore, TETA.4HCl is preferably in isolated form, i.e., TETA.4HCl is separated from any reaction mixture in which it is produced (usually purified) before starting crystallization to form B. In addition, crude TETA.4HCl is preferably recrystallized before starting the process to produce form B. This also provides a higher purity starting material and enables reliable production of form B crystals by following the methods described herein.

[0115] TETA.4HCl is usually dissolved in an aqueous solution before crystallization. Typically, the solution contains 0.01-10 g TETA.4HCl / per ml of water. Preferably, the solution contains 0.1-5 g TETA.4HCl / per ml of water, most preferably 0.6-1.2 g TETA.4HCl / per ml of water. The volume of the antisolvent used for recrystallization is usually 0.5 ml or less per 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, most preferably 0.01-0.1 ml / g TETA.4HCl.

[0116] Preferred crystallization methods for Form B include:

[0117] (i) adding an antisolvent to an aqueous solution of TETA.4HCl, preferably a stirred aqueous solution, at a time period t1 and / or at an addition rate R1 at a temperature T1;

[0118] (ii) optionally adding TETA.4HCl seeds;

[0119] (iii) optionally stirring the resulting mixture at T1 for a further period of time t1a;

[0120] (iv) optionally reducing the temperature to temperature T2 and stirring the mixture for a further period of time t2; and

[0121] (v) collecting the resulting crystals.

[0122] T1 is about 20°C or less, preferably about 15°C or less, more preferably about 10°C or less. To produce substantially pure Form B, T1 is preferably about 15°C or less, more preferably about 10°C or less.

[0123] 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 are preferred, particularly ethanol.

[0124] The solution is typically stirred or mixed during the addition, typically by agitation. Further agitation of the mixture, such as stirring, may be performed in steps (iii) and (iv).

[0125] The crystallization process is usually carried out over an extended period of time. Therefore, t1 is usually at least 1 hour, preferably at least 1.5 hours. The addition rate of the antisolvent R1 is usually 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.

[0126] If seed crystals are added, they are usually added before, during or after step (i). Preferably, seed crystals are added before step (i) or during the anti-solvent addition of step (i). If seed crystals are added during or after the anti-solvent is added, the mixture is usually stirred for an extended period of time t1a, 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 collecting the crystals. This further stirring step is carried out during t2 (preferably at least 30 minutes). The further stirring step is carried out at a reduced temperature T2. T2 is usually less than T1, preferably at least 5°C lower than T1, more preferably at least 10°C. T2 is usually about 10°C or lower, preferably about 5°C or lower, more preferably about 0°C. It has been found that increasing the time or rate of crystallization and reducing the temperature of crystallization can provide a higher purity of Type B. Introducing seed crystals also helps to improve the purity of Type B crystals.

[0127] In the above method, usually, 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. 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 less, 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 less, t1a is at least 4 hours, and t2 is at least 30 minutes.

[0128] The crystallization is preferably carried out under an inert atmosphere, for example under nitrogen.

[0129] A preferred crystallization method comprises at least steps (i), (iv) and (v) above. A more preferred method comprises steps (i), (iii), (iv) and (v). A most preferred method comprises all of steps (i) to (v). Thus, a preferred method for making TETA.4HCl comprises:

[0130] (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 an addition rate of R1;

[0131] (ii) optionally adding TETA.4HCl seeds;

[0132] (iii) optionally stirring the resulting mixture at T1 for a further period of time t1a;

[0133] (iv) reducing the temperature to temperature T2 and stirring the mixture for a further period of time t2; and

[0134] (v) collecting the resulting crystals.

[0135] 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 less, t1a is at least 3 hours, and t2 is at least 30 minutes.

[0136] Another preferred process suitable for producing substantially pure TETA.4HCl Form B comprises:

[0137] (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 an addition rate of R1;

[0138] (ii) adding TETA.4HCl seed crystals;

[0139] (iii) stirring the resulting mixture at T1 for a further period of time t1a;

[0140] (iv) reducing the temperature to temperature T2 and stirring the mixture for a further period of time t2; and

[0141] (v) collecting the resulting crystals.

[0142] 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 less, t1a is at least 4 hours, and t2 is at least 30 minutes.

[0143] The crystals may be collected by any suitable means, as long as the temperature of the collection step is maintained below about 40° C., preferably below about 30° C. It has been found that a higher temperature step performed before the crystals are completely dry can only form type A crystals. Most preferably, the collection of the crystals is performed at a temperature below about 25° C., such as about 20° C. or lower.

[0144] Suitable methods for collecting the crystals include filtration and centrifugation. Typically, the resulting crystals are dried at a temperature typically below about 40°C, preferably below about 30°C. Prior to drying, the crystals may be washed with, for example, an anti-solvent. Suitable anti-solvents for washing are those described above, particularly methanol or ethanol, most preferably ethanol. Drying is typically vacuum drying, as heating will result in the formation of Type A crystals. Preferably, vacuum drying is performed at a temperature below about 40°C.

[0145] If desired, the resulting dry product can be further processed, for example by grinding or granulation. Form B is substantially stable upon grinding.

[0146] Collection and further processing steps, such as washing, drying and grinding, where appropriate, are generally carried out under an inert atmosphere, for example under nitrogen.

[0147] Pharmaceutical compositions and dosage forms

[0148] The pharmaceutical composition of the present invention comprises the crystallized TETA.4HCl Form B described herein and 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 a tablet, capsule, powder, semisolid, sustained release formulation, solution, suspension or any other suitable composition. Tablets, capsules and powders (particularly tablets) are preferred.

[0149] In alternative embodiments, the composition is administered parenterally, for example subcutaneously or intravenously.

[0150] Pharmaceutical dosage forms can be prepared by subjecting the crystals produced as described herein to further processing steps. Thus, compositions (typically oral dosage forms) can be prepared in the following manner: (a) obtaining TETA.4HCl Type B, for example, using the above method, (b) optionally grinding and / or granulating the resulting crystals, (c) mixing TETA.4HCl Type B with a pharmaceutically acceptable carrier, and (d) optionally mixing TETA.4HCl Type B with a carrier. Suitable carriers are further described below. When the oral dosage form is a tablet, the method may further include (e) compressing the mixture to form a tablet, and optionally coating the tablet with sugar or film. Alternatively, the solid oral dosage form may be a capsule or a powder. In this case, the method of the present invention may further include (e) packaging the resulting mixture in, for example, a capsule. Other standard steps, such as grinding, granulation, sugar coating or film coating, may be included in the method.

[0151] The pharmaceutical composition typically comprises at most 85 wt% TETA.4HCl, for example at most 50 wt% TETA.4HCl.Preferred compositions are sterile and non-pyrogenic.

[0152] Suitable pharmaceutically acceptable carriers for preparing oral dosage forms include, for example, dissolving agents, such as cyclodextrins or modified cyclodextrins; diluents, such as lactose, dextrose, sucrose, cellulose, corn starch or potato starch; lubricants, such as silicon dioxide, talc, stearic acid, magnesium or calcium stearate and / or polyethylene glycol; binders, such as starch, tragacanth, gelatin, syrup, gum arabic, sorbitol, methylcellulose, carboxymethylcellulose or polyvinylpyrrolidone; disintegrators, such as starch, alginic acid, alginates or sodium starch glycolate; effervescent mixtures; dyes; sweeteners; wetting agents, such as lecithin, polysorbate, lauryl sulfate; and non-toxic and pharmacologically inactive substances commonly used in pharmaceutical preparations. Such pharmaceutical preparations can be produced in a known manner, for example by mixing,

[0153] The composition of the present invention generally comprises a high proportion of TETA.4HCl Form B and a small amount of Form A. Preferably, the pharmaceutical composition of the present invention comprises no more than 10 wt% of TETA.4HCl Form A. Preferably, the composition of the present invention is substantially free of triethylenetetraaminetetrachloride Form A. Substantially free of Form A as used herein means that the composition contains no more than 5 wt%, preferably no more than 2 wt%, more preferably no more than 1 wt% or 0.5 wt%, and most preferably no more than 0.1 wt% of TETA.4HCl Form A.

[0154] Medical Uses

[0155] A therapeutically effective amount of a compound of the invention is administered to a subject. It will be understood that the specific dosage level for any particular subject will depend on a variety of factors, including the activity of the particular compound used, age, body weight, general health, sex, diet, administration time, administration route, excretion rate, drug combination, and severity of the particular disease being treated. The optimal dosage level and frequency of administration will generally be determined by clinical trials.

[0156] Depending on the age, weight and condition of the subject to be treated, the type and severity of the disease, the frequency and route of administration, the usual daily dose is up to 50 mg per kg body weight, for example 0.001-50 mg per kg body weight. Preferably, the daily dose level is 0.05 mg-2 g, preferably 0.1 mg-10 mg. The compounds of the present invention are usually administered to patients in non-toxic amounts.

[0157] The invention also provides a crystalline form as defined herein or a composition as defined herein for use in a method of treating the human or animal body by therapy. In particular, the crystalline form and composition of the invention may be used to reduce the copper content of a subject and / or reduce the toxic effects of residual copper in a subject. Thus, they may be used to treat disorders and diseases associated with elevated copper levels. In particular, they may be used to prevent or treat Wilson's disease.

[0158] TETA itself is a known treatment for Wilson's disease and diseases and disorders associated with elevated copper levels. Upon administration of the crystalline form of the present invention, the compound will dissolve in the body system and the therapeutic effect of the crystalline form is expected to be the same as the known TETA form.

[0159] The subject treated according to the invention may be a human or an animal subject, particularly a human or a mammal, typically a human.

[0160] Example

[0161] Reference Example 1: Synthesis of crude TETA.4HCl

[0162] like Figure 1a and 1b As shown, crude TETA.4HCl was produced.

[0163] Example 2: Synthesis of TETA.4HCl Form B

[0164] according to Figure 2 The procedure shown produces substantially pure TETA.4HCl Form B.

[0165] Reference Example 3: Synthesis of TETA.4HCl Type A

[0166] Crude TETA 4HCl was dissolved in 2 volumes of pure water under nitrogen and the resulting solution was clarified by filtration. The reactor was heated to 70°C (jacket reactor temperature) and when the temperature of the reaction mixture was between 55-60°C, 7 volumes of methanol were added at a rate to maintain the temperature in the mass between 55-65°C to recrystallize the product.

[0167] After stirring for at least 30 minutes at 65° C. (reactor jacket temperature), the reaction mixture is slowly cooled over a period of at least 2 hours and 30 minutes, taking into account a cooling rate of about 5° C. per 30 minutes, so as to obtain a temperature in the mass between 30° C. and 35° C. The suspension is then stirred for at least 1 hour while the temperature in the mass is maintained between 30° C. and 35° C.

[0168] The product was filtered on an enameled Nutsch filter with 10 μm membrane pores and washed twice with 1.5 volumes of methanol. The last wash was followed by forced filtration to dry the product. The product was dried in a vacuum oven at 60° C. for at least 14 hours.

[0169] Example 4: Analysis of TETA.4HCl crystal form

[0170] X-ray powder diffraction

[0171] Several milligrams of the samples obtained in Example 2 and Reference Example 3 were placed between three polymer foils ( and polypropylene). The diffraction pattern showed a broad peak with a weak intensity near 2θ=5.5°.

[0172] The samples were placed in a PANALYTICAL X'PERT PRO MPD diffractometer configured in transmission mode and analyzed using the conditions shown in Table 1 below. The powder samples were exposed to a characteristic wavelength (λ) of Cu-K α Diffraction data were obtained using X-ray radiation. X-rays were generated from a Cu anode supplied with 40 kV and 40 mA current. Analyses were performed between 2° and 50° (unless otherwise stated). The calibration of the diffractometer was verified before each analysis.

[0173] Figure 3 The XRPD pattern of a mixture of Form A and Form B ("curve 1+2") and the XRPD pattern of crystals produced according to Reference Example 3 (substantially pure Form A: "curve 1") are shown. The arrows mark peaks unique to Form B. Figure 4 An XRPD pattern of crystals produced according to Example 2 (substantially pure Form B) is shown.

[0174] Table 1: XRPD analysis conditions

[0175]

[0176]

[0177] Raman spectroscopy

[0178] The samples of Example 2 and Reference Example 3 were analyzed by Raman spectroscopy. A Renishaw RA802 pharmaceutical analyzer was used under the following conditions:

[0179] Table 2:

[0180] Laser wavelength 785nm Spectral dispersion <![CDATA[2cm -1 / pixel]]> Objective Hi Mag (x50–1μm capability) Focus Automatic (real-time track) Collection time 1s Laser power 50％

[0181] Spectrum Figure 5a and 5b In Figure 5b In the figure, the lower line represents Example 2 (Form B), while the upper line represents Reference Example 3 (Form A). The arrows depict the shifts that are characteristic of Form B. The Raman spectrum of Example 2 shows 943, 1173, 1527 and 1612 cm -1 The Raman spectrum of Reference Example 3 (Type A) has bands at 933, 1167, 1513 and 1604 cm -1 Peaks are shown.

[0182] FTIR-ATR analysis

[0183] Infrared spectra were measured on a Nicolet iS5 FT-IR spectrometer equipped with an iS7ATR module with the following parameters set:

[0184] Table 3:

[0185] model ATR Diamond Resolution <![CDATA[4cm -1 ]]> Scan times (measurement) 32scans Scan times (background) 32scans spectrum <![CDATA[4000cm -1 Up to 525cm -1 Absorbance]]>

[0186] An infrared spectrum was obtained for Example 2 (Form B). The spectrum shows Figure 8a and b.

[0187] DSC analysis

[0188] DSC analysis was performed using a Mettler Toledo DSC3+ (serial number B531255222) in a 40 μl sealed aluminum pan under nitrogen blowing at 50 mL / min, and the lid was perforated prior to analysis.

[0189] Table 4:

[0190] Sample ID Example 2 Sample weight 0.360mg Scan range 20℃–300℃ Scan rate 10° / min

[0191] An endothermic event corresponding to melting of the sample was observed. The onset and peak temperatures are shown in the table below.

[0192] Table 5:

[0193] T Start T peak Enthalpy Remark (℃) (℃) (J / g) 252.1 259.2 232.1 Melting

[0194] Onset temperature, peak temperature and enthalpy

[0195] Thermogravimetric Analysis (TGA)

[0196] Thermogravimetric analysis was performed using a Pyris 1 TGA analyzer (serial number 537N7052501) under nitrogen blowing at 20 mL / min in sealed aluminum pans that were perforated prior to analysis.

[0197] Table 6:

[0198] Sample ID Example 2 Sample weight 5.100mg Starting temperature 25℃ End temperature 300℃ Scan rate 10℃ / min

[0199] Thermogravimetric analysis showed that the weight loss started at 225 °C, which increased after 289.5 °C. This may be due to degradation. The weight loss was 10.24%: 2.64% at 225.0-289.5 °C and 7.60% at 289.5-299.0 °C

[0200] Dynamic Vapour Sorption (DVS) Analysis

[0201] DVS analysis was performed using a SMSDVS Intrinsic Analyzer (Serial No. PF 140088) in an open aluminum pan at 25°C with a nitrogen purge gas of 100 ml / min. The stability criterion was a weight change of less than 0.002% over a 5 minute time frame (minimum 10 minutes, maximum 100 minutes).

[0202] Table 7:

[0203]

[0204] Fig.11a (Example 2) and 11b (Reference Example 3) provide DVS isotherm plots. DVS analysis performed on these two samples showed a significant weight increase at high relative humidity values ​​(90% RH and above):

[0205] - For Example 2, at least +55.8%

[0206] -For Reference Example 3, at least +60.8%

[0207] The desorption phases show different behavior for the two solids. Example 2 almost reaches its reference weight in the second desorption phase. For reference example 3, at the end of the second desorption phase, the weight is still +14.4% higher than the reference weight (still 7.9% higher for the minimum value reached). It is noteworthy that for the latter, these steps have reached the time limit criteria (hence, the weight of the sample is not stable).

[0208] Example 5: Heckel test

[0209] 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 known weight of material is compressed in a die with a diameter of 10 mm and a flat punch is moved 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 component of the material deformation. The samples produced according to Example 2 and Reference Example 3 were subjected to the Heckel test.

[0210] method

[0211] The true density was determined by helium pychnometry.

[0212] Equipment used: Micromeritics AccuPyc II 1340

[0213] Test parameters:

[0214] Cup size: 3.5cm3

[0215] Purge times 5

[0216] Purge pressure 19.5 psig

[0217] Run times 10

[0218] Operating fill pressure 19.5 psig

[0219] Balance rate 0.02psig

[0220] The operating accuracy is

[0221] Full scale percentage 0.05%

[0222] Tests were performed in duplicate. (Assuming a target of <2% variability was achieved).

[0223] compression

[0224] A known weight of pure drug was compacted to theoretical zero porosity using a flat punch with a diameter of 10 mm. A compaction simulator was used under the following conditions:

[0225] Tool: 10mm round flat surface

[0226] Profile: V-shaped profile

[0227] Punch speed – slower by 0.1 mm / s

[0228] Punch speed – 300mm / s faster

[0229] Die lubrication: Yes, using magnesium stearate in acetone

[0230] Repetitions: 3

[0231] - Elasticity correction: Yes

[0232] During the compression process, the position of the punch and the measured force can be accurately determined by the load cell, resulting in a record of the key compression parameters. Temperature and humidity are monitored regularly during the test. The data is analyzed by the Compaction Analysis software program to generate the yield stress (Py) value using the Heckel equation:

[0233]

[0234] Where D = relative density of the briquette

[0235] P = applied pressure

[0236] K = slope of the line in the linear region

[0237] [Reference: RW Heckel. Trans. Metall. Soc. AIME 221 (1961) 1001-1008]

[0238] Strain Rate Sensitivity (SRS)

[0239] For some materials, the deformation characteristics change as a function of the rate at which the force is applied. This can be estimated by calculating the strain rate sensitivity. The yield pressure at high compression speeds is compared to the yield pressure at low compression speeds using the following formula:

[0240]

[0241] [Reference: RJRoberts and RCRoe, Chem.Eng.Sci.42(1987)p903].

[0242] result

[0243] True density

[0244] Table 8:

[0245]

[0246] *The difference between Runs 1 and 2 exceeded 2%. A third run was performed. The data from Run 2 were assumed to be outliers and were not used in the mean calculation.

[0247] Table 9: Briquetting results: Reference Example 3

[0248]

[0249] Laboratory conditions: 21.3℃ / 52.1%RH

[0250] Table 10: Briquetting results: Reference Example 3

[0251]

[0252] Laboratory conditions: 21.6℃ / 50.0% RH

[0253] Table 11: Summary of Results and Observations

[0254] Batch Processing Properties Reference Example 3 Yield pressure slow (0.1mm / s) 98.17Mpa(±3.07) Fast yield pressure (300mm / s) 127.42Mpa(±2.23) Strain rate sensitivity 29.8％

[0255] Table 12: Briquetting Results: Example 2

[0256] Slow speed 0.1mm / s Run 1 Run 2 Run 3 Run 4 Yield pressure(Mpa) 106.465 117.516 118.116 115.227 Calculation linear range (Mpa) 25-150 25-150 25-150 25-150 Upper punch peak force (kN) 20.211 24.088 24.175 23.906 Demolding force(kN) 0.044 0.053 0.080 0.060 Briquetting observation: Good gloss tablets Good gloss tablets Good gloss tablets Good gloss tablets Kiloponds 12.07 7.52 9.64 9.19

[0257] Laboratory conditions: 21.7℃ / 51.1%RH

[0258] Table 13: Briquetting Results: Example 2

[0259]

[0260] Laboratory conditions: 21.7℃ / 51.2%RH

[0261] Table 14: Summary of Results and Observations

[0262] Batch Processing Properties Example 2 Yield pressure slow (0.1mm / s) 114.33Mpa(±5.39) Fast yield pressure (300mm / s) 129.61Mpa(±0.44) Strain rate sensitivity 13.4％

[0263] The Heckel test is a measure of the deformation of the tablet formed. The briquette strength is an indication of how the residence time affects the cohesion of the briquette. It was found that the briquette produced by Reference Example 2 at a slow speed had a moderate tensile strength (6-11 kilograms of force (kiloppond)). At a fast speed, the strength was reduced to about 4-6 kilograms of force. On the other hand, Example 3 showed a strength of 7-12 kilograms of force at a slow speed, but showed a strength of 9-11 kilograms of force at a fast speed, indicating that Example 3 has a higher tensile strength of the briquette product under a fast briquetting process. Figure 6 The strain rate sensitivity of Example 2 and Reference Example 3 at different production speeds is shown.

[0264] Example 6: Aging of tablets

[0265] The TETA.4HCl obtained according to Reference Example 3 was compressed into tablets. The images of the tablets are shown in Figure 7a The tablets were aged at 40°C and 75% humidity for six months. After aging, the tablets were observed to have many discolored patches. Figure 7b Images of aged tablets are provided in .

[0266] The tablets were analyzed by Raman spectroscopy under the same conditions as described in Example 4 above, and the results were compared with the Raman spectra of Form A and Form B TETA.4HCl. Figure 7c Areas of the tablet showing the presence of TETA.4HCl Form A are shown in the darker areas. Figure 7b and Figure 7c It is clear from the comparison that the area of ​​TETA.4HCl Form A on the tablet surface correlates with the location of the discolored area.

[0267] Trace amounts of TETA.4HCl Form B were detected in the tablets, which were probably formed under compression of the TETA.4HCl forming the tablets. The areas of TETA.4HCl Form B did not correlate with the discolored areas observed in the aged tablets.

[0268] Example 7: Crystallization process

[0269] Crystallization tests for the preparation of Form B were performed at 20°C. The starting solution was prepared in a mixture of ethanol / water at (25:75) and ethanol was added until a (75:25) ratio was reached. The first test was performed by manually adding ethanol. Additional tests were then performed using a syringe pump, adding at a slow and controlled rate.

[0270] Manual addition (drop by drop)

[0271] In these tests, the addition was done manually using a micropipette. A solution of TETA.4HCl in a (25:75) ethanol / water mixture was placed under agitation and thermostated at 20°C. Table 15 lists the amounts of TETA.4HCl starting material.

[0272] Antisolvent (ethanol) was added dropwise regularly. Two addition "rates" were tested. After adding the antisolvent, the solid phase was sampled and analyzed by X-ray diffraction to determine the properties of the solid phase. The conditions of X-ray diffraction are as described in Example 4. The last test was carried out at the beginning of the addition, with a slower addition rate when type B seeds appeared (PE1716E007-L-5). This test resulted in a solid phase in which the signal of type A was not observed on the diffraction pattern.

[0273] The results are listed in Table 15 and the diffraction pattern obtained after analyzing the solid phase in the suspension at the end of the ethanol addition is shown in Figure 9a , b and c. Figure 9a -c shows the XRPD diffraction pattern, starting from the bottom line:

[0274] Reference for Type A ( Figure 9a -c in the bottom line)

[0275] Reference for Type B

[0276] PE1716E007-L-5

[0277] PE1716E007-L-4

[0278] PE1716E007-L-3

[0279] PE1716E007-L-2

[0280] PE1716E007-L-1

[0281] PE1716E007-R-1( Figure 9a -c at the top of the line)

[0282]

[0283] Programmatic addition (continuous)

[0284] In this series of tests, the addition of antisolvent was driven by a syringe pump. This enabled continuous addition at a very low rate. The same protocol as above was used: a solution of the starting material was prepared nearly saturated in a 25:75 ethanol / water mixture as shown in Table 16. The solution was then saturated with Form B until the solid phase remained in suspension: this ensured that seeds of Form B were present at the start of antisolvent addition.

[0285] Two addition rates were tested: 0.05 mL / min and 0.1 mL / min (for 750 mg of starting material in solution). Ethanol was added at these selected rates until an ethanol / water ratio of 75:25 was obtained. Two additional tests were performed, adding up to 82:18 (equivalent to a concentration of about 12.5%) and adding up to 87.5 / 12.5 (equivalent to a concentration of about 9%) (starting material / total weight).

[0286] X-ray diffraction analysis was performed using the conditions listed in Example 4. The results are shown in Table 16 below, and the diffraction pattern obtained after analyzing the solid phase in the suspension at the end of the ethanol addition is shown in Figure 10a-10cThese show the diffraction curves for two tests with final ethanol / water ratios of 75:25 and 87.5:12.5 where no signal for Form A was observed. For the test with 82:18, there is a small shoulder to the left of the peak at 2θ = 25.4° (corresponding to Form A).

[0287] Fig.10a -c shows the XRPD diffraction pattern, starting from the bottom line:

[0288] Reference for Type A ( Fig.10a -c bottom line)

[0289] Reference for Type B

[0290] PE1716E007-L-6

[0291] PE1716E007-L-7

[0292] PE1716E007-L-8

[0293] PE1716E007-L-9( Fig.10a -c at the top of the line)

[0294]

[0295] Example 8: Crystallization process

[0296] The following scheme shows an overview of the synthesis of triethylenetetramine (triethylenetetramine) tetrahydrochloride (TETA 4HCl).

[0297]

[0298] In step I, triethylenetetramine (TETA) is converted into the corresponding triethylenetetramine hydrate (TETA hydrate) by stirring in the presence of water and TBME. Isopropanol is added as an anti-solvent and, if necessary, TETA hydrate is used as a seed. TETA hydrate is crystallized, filtered and separated. Crude triethylenetetramine tetrahydrochloride (crude TETA 4HCl) is obtained by reacting triethylenetetramine hydrate (TETA hydrate) with aqueous hydrochloric acid in ethanol (step II). Crude triethylenetetramine tetrahydrochloride (crude TETA 4HCl) is recrystallized from a mixture of pure water and ethanol. In the presence of type B seed crystals, crude triethylenetetramine tetrahydrochloride (crude TETA4HCl) is further purified by recrystallization from a mixture of pure water and ethanol to obtain triethylenetetramine tetrahydrochloride (TETA 4HCl) (step III).

[0299] The process produces 110-130 kg of TETA 4HCl in batches from 125 kg of TETA. The overall yield of the synthesis of crude TETA 4HCl including two recrystallizations is about 50%.

[0300] like Figure 1a , 1b As shown in the flow chart of FIG. 2, recrystallization is performed to produce type B crystals, but in step II ( Figure 1b ) and Step III ( Figure 2 ), crude TETA.4HCl was added for further recrystallization. The process can be described as follows:

[0301] Once the equipment had been inerted, all manipulations were performed under a nitrogen flow.

[0302] Step I: Preparation of triethylenetetramine hydrate (TETA hydrate)

[0303] • Triethylenetetramine (nominal amount 125 kg) was charged into the reactor, followed by TBME (185 ± 5 kg).

[0304] Water (about 28 kg) was added and agitated for ≥ 15 minutes while maintaining the temperature at ≤ 30 °C.

[0305] If necessary, inoculate the solution with triethylenetetramine hydrate (about 0.1 kg) while stirring at 25-35°C.

[0306] To promote crystallization.

[0307] • Add isopropanol (64 ± 1 kg) at 25-35 °C.

[0308] The suspension was heated at 30-40°C for ≥15 minutes and then slowly cooled to 15-25°C over a period of ≥90 minutes.

[0309] Cool the suspension to -5-5°C and stir for ≥30 minutes

[0310] The product was filtered and centrifuged. Then, a sample was taken for analysis (GC determination) and determination of impurities.

[0311] If the sample is pure enough, load the wet TETA hydrate into a dryer and dry it at a temperature of ≤25°C.

[0312] Until the requirements of the next step are met.

[0313] The dried product was analyzed for purity by GC, appearance and residual water by KF, composition by FTIR, and sulfated ash and residual solvent by GC.

[0314] Step II: Production of crude triethylenetetramine tetrahydrochloride (crude TETA.4HCl)

[0315] Dissolve triethylenetetramine hydrate (TETA hydrate) in water (85±1 kg) and acidify with concentrated aqueous hydrochloric acid (200±5 kg) fed over ≥1 hour at ≤40°C. Check pH (target pH=1.0) and add concentrated aqueous hydrochloric acid until pH≤1.0.

[0316] - Cool the reaction mixture to 15-25 °C and stir for ≥ 10 minutes.

[0317] Treat the solution with ethanol (672 ± 5 kg) for ≥ 1.5 h, and maintain the temperature at ≤ 30 °C.

[0318] The suspension was cooled to -5-5°C and stirred for ≥30 minutes.

[0319] - The product was filtered and the solid was washed sequentially with ethanol (1 x 20 kg, then 3 x 25 kg).

[0320] Dissolve crude triethylenetetramine tetrahydrochloride (crude TETA.4HCl) in water (340±10 kg).

[0321] Treat the solution with ethanol (909 ± 10 kg) for ≥ 1.5 hours, maintaining the temperature at ≤ 30 °C.

[0322] The suspension was cooled to -5-5°C and stirred for ≥30 minutes.

[0323] - The product was filtered and the solid was washed sequentially with ethanol (1 x 14 kg, then 3 x 15 kg).

[0324] Step III: Production of triethylenetetramine tetrahydrochloride (TETA.4HCl)

[0325] Crude triethylenetetramine tetrahydrochloride (crude TETA.4HCl) was dissolved in water (340±10 kg).

[0326] - Treat the solution with ethanol (909 ± 15 kg) over a period of ≥ 1.5 hours, maintaining the temperature at 7-13 °C.

[0327] - During the addition of ethanol, the solution was seeded with TETA.4HCl (2 wt%).

[0328] The suspension was stirred for ≥5 hours, then cooled to -5°C and stirred for ≥30 minutes.

[0329] - The product was filtered and the solid was washed sequentially with ethanol (1 x 14 kg, then 3 x 15 kg).

[0330] • The purity of product samples was analyzed by GC.

[0331] The product is dried at a temperature of ≤40°C and ground if the control parameters of loss on drying are met.

[0332] The ground API was transferred into a double-layer food-grade polyethylene bag under nitrogen, then placed into an aluminum bag and sealed. The aluminum bag was inserted into the HDPE drum.

[0333] Reprocessing

[0334] The TETA 4HCl 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 triethylenetetramine tetrahydrochloride type B, having at least one of the following characteristics: (i) an XRPD pattern having at least two peaks selected from peaks at 22.9, 25.4, 25.8, 26.6, 34.6, and 35.3 ± 0.1° 2θ, and (ii) With the range from 943, 1173, 1527 and 1612 ± 5 cm -1 a Raman spectrum of at least two peaks selected from peaks at Raman shifts of wherein the crystalline form comprises no more than 50 wt% of triethylenetetramine tetrahydrochloride Form A, the triethylenetetramine tetrahydrochloride Form A having an XRPD pattern having peaks at 25.2 and 35.7 ± 0.1 ° 2θ, and the peaks of the XRPD pattern are Wavelength measurement.

2. The crystalline form of claim 1 having an XRPD pattern having peaks at 25.4, 34.6 and 35.3 ± 0.1° 2θ.

3. The crystalline form of claim 1, comprising no more than 40 wt% of triethylenetetramine tetrahydrochloride Form A, wherein the triethylenetetramine tetrahydrochloride Form A has an XRPD pattern having peaks at 25.2 and 35.7 ± 0.1° 2θ.

4. The crystalline form of claim 1, comprising no more than 20 wt% of triethylenetetramine tetrahydrochloride Form A, wherein the triethylenetetramine tetrahydrochloride Form A has an XRPD pattern having peaks at 25.2 and 35.7 ± 0.1° 2θ.

5. The crystalline form of claim 1, consisting essentially of triethylenetetramine tetrahydrochloride Form B, wherein the triethylenetetramine tetrahydrochloride Form B has: (i) an XRPD pattern as defined in claim 1; and / or (ii) With the range from 943, 1173, 1527 and 1612 ± 5 cm -1 A Raman spectrum of at least two peaks selected from the peaks at the Raman shift.

6. A pharmaceutical composition comprising the crystalline form according to claim 1 and a pharmaceutically acceptable carrier or diluent.

7. The pharmaceutical composition of claim 6, comprising no more than 10 wt% of triethylenetetramine tetrahydrochloride Form A, said triethylenetetramine tetrahydrochloride Form A having an XRPD pattern having peaks at 25.2 and 35.7 ± 0.1° 2θ.

8. The pharmaceutical composition of claim 6, comprising no more than 5 wt% of triethylenetetramine tetrahydrochloride Form A, wherein the triethylenetetramine tetrahydrochloride Form A has an XRPD pattern having peaks at 25.2 and 35.7 ± 0.1° 2θ.

9. A solid oral dosage form composition comprising the crystalline form according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

10. A pharmaceutical composition comprising crystalline triethylenetetramine tetrahydrochloride, wherein at least 50 wt% of the crystalline triethylenetetramine tetrahydrochloride is Form B and no more than 50 wt% of the crystalline triethylenetetramine tetrahydrochloride is Form A, wherein: The type B has at least one of the following characteristics: (i) an XRPD pattern having at least two peaks selected from peaks at 22.9, 25.4, 25.8, 26.6, 34.6 and 35.3 ± 0.1° 2θ, and, (ii) With the range from 943, 1173, 1527 and 1612 ± 5 cm -1 a Raman spectrum of at least two peaks selected from peaks at Raman shifts of The Form A has an XRPD pattern with peaks at 25.2 and 35.7 ± 0.1° 2Θ.

11. The pharmaceutical composition according to claim 10, wherein no more than 40 wt% of the crystalline triethylenetetramine tetrahydrochloride is the A form.

12. The pharmaceutical composition according to claim 11, wherein no more than 20 wt% of the crystalline triethylenetetramine tetrahydrochloride is the A form.

13. Use of the crystalline form according to any one of claims 1 to 5, the pharmaceutical composition according to any one of claims 6 to 8 or 10 to 12, or the solid oral dosage form composition according to claim 9 in the preparation of a medicament for preventing or treating Wilson's disease.

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