MANUFACTURING PROCESS FOR AMIFAMPRIDINE PHOSPHATE

Abstract

The present invention relates to a crystalline form of amifampridine phosphate having an irregular or plate-like, not needle-like crystal habit and to a process for its manufacture.

Claims

1. Phosphate salt of amifampridine, wherein the salt is obtainable by a process comprising the steps: i) preparing an alcoholic solution containing amifampridine, wherein the solvent is an alcohol optionally in admixture with not more than 10 wt % water, ii) precipitating the phosphate salt of amifampridine by adding an aqueous phosphoric acid solution to the alcoholic solution obtained in step (i), wherein the aqueous phosphoric acid solution contains at least 70 wt % phosphoric acid, and iii) isolating the phosphate salt of amifampridine obtained in step (ii).

2. The salt according to claim 1, characterized by the following peaks of the powder X-ray diffractogram (Cu-Kα1=1.54059 Å): 11.06, 12.16, 12.69, 12.91, 15.45, 17.12, 17.42, 18.07, 20.21, 21.60, 22.22, 22.57, 22.95, 24.03, 24.59, 25.11, 25.54, 26.05, 26.69, 27.02, 27.76, 28.21, 28.69, 29.54, 30.19, 30.27 and 30.44±0.20° 2θ.

3. The salt according to claim 2, wherein the powder X-ray diffractogram does not contain an additional peak in the range of 5.00 to 30.44±0.20° 2θ.

4. The salt according to claim 1, characterized by the following peaks of the powder X-ray diffractogram (Cu-Kα1=1.54059 Å): 15.5, 17.4, 25.5 and 26.1±0.20° 2θ.

5. The salt according to claim 2 or 4, wherein the powder X-ray diffractogram does not contain peaks at 8.39, 16.16, 19.69 and 21.16±0.20° 2θ.

6. A process for preparing a phosphate salt of amifampridine, wherein the process comprises the steps: i) preparing an alcoholic solution containing amifampridine, wherein the solvent is an alcohol optionally in admixture with not more than 10 wt % water, ii) precipitating the phosphate salt of amifampridine by adding an aqueous phosphoric acid solution to the alcoholic solution obtained in step (i), wherein the aqueous phosphoric acid solution contains at least 70 wt % phosphoric acid, and iii) isolating the phosphate salt of amifampridine obtained in step (ii).

7. The process according to claim 6, wherein the solvent is ethanol optionally in admixture with not more than 10 wt % water.

8. The process according to claim 7, wherein the solvent is ethanol in admixture with not more than 5 wt % water.

9. The process according to any one of claims 6 to 8, wherein the aqueous phosphoric acid solution contains at least 80 wt %, preferably at least 85 wt % phosphoric acid.

10. A process for preparing a solid unit dosage form for oral administration containing a phosphate salt of amifampridine, wherein the process comprises the steps: i) preparing an alcoholic solution containing amifampridine, wherein the solvent is an alcohol optionally in admixture with not more than 10 wt % water, ii) precipitating the phosphate salt of amifampridine by adding an aqueous phosphoric acid solution to the alcoholic solution obtained in step (i), wherein the aqueous phosphoric acid solution contains at least 70 wt % phosphoric acid, iii) isolating the phosphate salt of amifampridine obtained in step (ii), iv) mixing the phosphate salt of amifampridine obtained in step (iii) and a pharmaceutical excipient to obtain a powdery blend, and v) converting the powdery blend obtained in step (iv) into the solid unit dosage form.

11. The process according to claim 10, wherein the solvent is ethanol optionally in admixture with not more than 10 wt % water.

12. The process according to claim 11, wherein the solvent is ethanol in admixture with not more than 5 wt % water.

13. The process according to any one of claims 10 to 12, wherein the aqueous phosphoric acid solution contains at least 80 wt %, preferably at least 85 wt % phosphoric acid.

14. The process according to any one of claims 10 to 13, wherein the solid unit dosage form is an optionally film-coated tablet.

15. The process according to claim 14, wherein step (v) comprises a) subjecting the powdery blend obtained in step (iv) to compression to obtain the tablet, or b1) subjecting the powdery blend obtained in step (iv) to compaction, b2) milling the compacted material obtained in step (b1) to obtain granules, b3) optionally mixing the granules obtained in step (b2) and a pharmaceutical excipient to obtain a mixture, and b4) subjecting the granules obtained in step (b2) or the mixture obtained in step (b3) to compression to obtain the tablet.

Description

FIGURES

[0019] FIG. 1: XRPD of form 1

[0020] FIG. 2: XRPD of form 1 in admixture with a small amount of form 2 (about 15%)

[0021] FIG. 3: Microscopic image of form 1 obtained by the process of the present invention in silicone oil

[0022] FIG. 4: Microscopic image of form 1 obtained by the process of the present invention in silicone oil

[0023] FIG. 5: Microscopic image of form 1 with form 2 (about 15%) obtained by the process of the present invention in silicone oil

[0024] FIG. 6: Microscopic image of form 1 obtained by the prior art method in silicone oil

[0025] The polymorph form 1 of 3,4-DAPP is known from the prior art, for example the FIG. 1 of Powder Diffraction 2011, 26(4), 321-325, and FIG. 5 of Cryst. Growth Des. 2013, 13, 708-715, are showing powder XRD patterns of pure form 1 with essentially the same characteristic peaks as the pattern of the crystalline form 1 in the FIG. 1 of the current application. Also the simulated powder XRD patterns based on the single crystal data disclosed in these prior art publications are essentially the same as the pattern in the FIG. 1 of the present application.

[0026] The inventors have analyzed the polymorphic form 1 of amifampridine phosphate and find the characteristic peaks to be present at 11.1, 15.5, 17.1, 17.4, 22.6, 24.6, 25.5 and 26.1*0.2° 2θ in a powder X-ray diffraction pattern (FIG. 1). Preferably, characteristic peaks for form 1 are 11.1, 15.5, 17.4, 25.5 and 26.1±0.2° 2θ. More preferably, characteristic peaks for form 1 are the strong reflexes at 15.5, 17.4, 25.5 and 26.1±0.2° 2θ with high relative intensity.

[0027] The salt of the present invention is characterized by the following detailed list of peaks of the powder X-ray diffractogram of form 1 (Cu-Kα1=1.54059 Å): 11.06, 12.16, 12.69, 12.91, 15.45, 17.12, 17.42, 18,07, 20.21, 21.60, 22.22, 22,57, 22.95, 24.03, 24.59, 25.11, 25.54, 26.05, 26,69, 27.02, 27.76, 28.21, 28,69, 29,54, 30.19, 30.27 and 30.44±0.20° 2θ. The polymorphically pure form 1 of amifampridine phosphate does not contain any additional peak in the range of 5.00 to 30.44±0.200° 2θ. The complete powder XRD-pattern of form 1 is shown in table 1 below.

TABLE-US-00001 TABLE 1 Form 1 No. Pos. [°2θ] Relative Intensity [%] 1 11.0668(3) 6.21 2 12.1645(9) 2.23 3 12.6903(3) 5.59 4 12.9147 0.52 5 15.4506(1) 63.77 6 17.1273(2) 22.46 7 17.4226(1) 100 8 18.0740(5) 7.22 9 20.2130(7) 3.24 10 21.6028(6) 5.04 11 22.2150(6) 6.84 12 22.5749(4) 13.16 13 22.9486(6) 6.98 14 24.0290(6) 4.54 15 24.5867(3) 17.17 16 25.113(1) 2.35 17 25.5379(2) 50.19 18 26.0493(1) 75.88 19 26.6906(5) 7.6 20 27.023(1) 3.63 21 27.7553(4) 7.48 22 28.2138(2) 5.11 23 28.6898(4) 2.03 24 29.5430(1) 13.56 25 30.19(1) 2.64 26 30.271(6) 3.45 27 30.4422(4) 18.46 28 31.184(2) 2.34 29 31.352(1) 3.97 30 32.1460(2) 3.74 31 32.78(4) 1.42 32 32.88(7) 0.85 33 33.4885(3) 3.33 34 33.784(1) 0.79 35 34.6712(4) 1.84 36 35.15(2) 2.6 37 35.27(1) 2.71 38 35.430(6) 3.19 39 35.9778(3) 2.8 40 36.586(2) 1.88 41 36.6640(8) 0.68 42 36.932(7) 1.02 43 37.33(3) 0.06 44 37.531(1) 0.93 45 39.050(1) 1.08 46 39.4071(5) 3.75 47 39.628(1) 1.32

[0028] Polymorphically pure form 1 of amifampridine phosphate is further characterized by a PXRD pattern substantially the same as the pattern shown in FIG. 1.

[0029] Polymorphically pure form 1 is obtained if the phosphate salt of amifampridine is precipitated from an ethanolic solution containing amifampridine by adding an aqueous phosphoric acid solution. A mixture of the form 1 and the form 2 is obtained if amifampridine is dissolved in methanol, optionally in admixture with not more than 10 wt % water, and precipitating the phosphate salt of amifampridine by adding an aqueous phosphoric acid solution.

[0030] The complete powder XRD-pattern of form 1 with a minor amount of form 2 (about 15%) is shown in table 2 below. Approximate estimate of 15% content of form 2 is based on the integral ratio of all peaks of known form 1 versus all peaks of form 2.

TABLE-US-00002 TABLE 2 Form 1 with form 2 No. Pos. [°2θ] Relative Intensity [%] 1 8.046(7) 0.51 2 8.392(6) 1.44 3 8.61(1) 0.7 4 8.918(6) 0.54 5 10.139(6) 0.18 6 10.499(2) 0.47 7 11.0814(3) 7.54 8 12.1766(7) 3.11 9 12.7021(7) 3.4 10 12.941(6) 0.32 11 13.129(8) 0.34 12 13.478(1) 0.2 13 14.908(2) 0.19 14 15.4655(2) 72.12 15 16.1589(7) 2.19 16 17.1417(4) 21.24 17 17.4436(2) 100 18 18.0805(4) 7.73 19 18.7404(8) 0.86 20 19.0350(8) 0.68 21 19.2946(5) 1.17 22 19.6938(3) 2.41 23 20.1998(2) 3.22 24 21.1577(6) 1.13 25 21.5871(2) 5.22 26 22.2162(1) 8.11 27 22.5725(1) 13.24 28 22.9410(2) 4.31 29 24.0655(2) 5.83 30 24.5829(1) 12.34 31 25.2020(8) 2.88 32 25.53686(8) 34.19 33 26.04124(5) 56.84 34 26.6816(1) 9.58 35 27.0425(2) 4.19 36 27.7753(2) 5.61 37 28.2320(2) 5.45 38 28.7014(5) 1.59 39 29.5568(1) 11.24 40 30.260(1) 4.95 41 30.4604(3) 13.87 42 31.201(3) 2.63 43 31.370(1) 4.44 44 32.1541(3) 2.73 45 32.778(3) 0.76 46 32.876(1) 1.2 47 33.5111(3) 3.08 48 33.795(1) 0.47

[0031] The comparison of table 1 with table 2 as well as the FIG. 1 with FIG. 2 leads to the conclusion that the most suitable peaks for the monitoring of the form 2 presence in the amifampridine phosphate API are the characteristic and non-interfering reflexes at 8.39, 16.16, 19.69, and 21.16±0.2° 2θ.

[0032] In some embodiments, one or more additional reflexes at 8.05, 8.61, 8.92, 18.74, 19.29, and 21.16°±0.2° 2θ, which are also characteristic, non-interfering and well-visible peaks in the XRPD pattern despite signals from form 1, may be employed for the detection of the presence of crystalline form 2 as polymorphic impurity.

[0033] The powder X-ray diffraction pattern of the polymorphically pure form 1 of amifampridine phosphate API is showing only characteristic peaks of the polymorphic form 1, for example at 15.5, 17.4, 25.5 and 26.1±0.2° 2θ, whilst no characteristic peaks are observed for form 2, for example at 8.39, 16.16, 19.69, and 21.16±0.2° 2θ, and optionally one or more additional reflexes at 8.05, 8.61, 8.92, 18.74, 19.23 and 21.16°±0.2° 2θ.

[0034] The present invention further relates to a process for preparing a phosphate salt of amifampridine, wherein the process comprises the steps: [0035] i) preparing an alcoholic solution containing amifampridine, wherein the solvent is an alcohol optionally in admixture with not more than 10 wt/n water, [0036] ii) precipitating the phosphate salt of amifampridine by adding an aqueous phosphoric acid solution to the alcoholic solution obtained in step (i), wherein the aqueous phosphoric acid solution contains at least 70 wt % phosphoric acid, and [0037] iii) isolating the phosphate salt of amifampridine obtained in step (ii).

[0038] Examples of suitable alcohols include methanol, ethanol, n-propanol and iso-propanol. Preferably, the solvent is ethanol optionally in admixture with not more than 10 wt % water. In a preferred embodiment, the solvent is ethanol in admixture with not more than 5 wt % water.

[0039] The addition of the aqueous phosphoric acid solution to the alcoholic solution is preferably conducted at elevated temperature, e.g. at 50-70° C. if an ethanolic solution is used (cf. Example 1). Heating the alcoholic solution allows the preparation of much higher concentrated solutions and, thus, saves solvent and, furthermore, allows to prepare larger batch sizes of the active drug ingredient in the same reactor.

[0040] Preferably, the aqueous phosphoric acid solution contains at least 80 wt %, more preferably at least 85 wt % phosphoric acid.

[0041] The phosphate salt of amifampridine obtained by the process of the present invention can be directly processed into a solid unit dosage form for oral administration, i.e. without further purification and without micronization. Hence, the present invention also relates to a process for preparing a solid unit dosage form for oral administration containing a phosphate salt of amifampridine, wherein the process comprises the steps: [0042] i) preparing an alcoholic solution containing amifampridine, wherein the solvent is an alcohol optionally in admixture with not more than 10 wt % water, [0043] ii) precipitating the phosphate salt of amifampridine by adding an aqueous phosphoric acid solution to the alcoholic solution obtained in step (i), wherein the aqueous phosphoric acid solution contains at least 70 wt % phosphoric acid, [0044] iii) isolating the phosphate salt of amifampridine obtained in step (ii), [0045] iv) mixing the phosphate salt of amifampridine obtained in step (iii) and a pharmaceutical excipient to obtain a powdery blend, and [0046] v) converting the powdery blend obtained in step (iv) into the solid unit dosage form.

[0047] Preferably, the solvent is ethanol optionally in admixture with not more than 10 wt % water. In a preferred embodiment, the solvent is ethanol in admixture with not more than 5 wt % water.

[0048] It is also preferred that the aqueous phosphoric acid solution contains at least 80 wt %, more preferred at least 85 wt/o phosphoric acid.

[0049] The solid unit dosage form may be an optionally film-coated tablet. The tablet may be prepared by a process, wherein step (v) comprises [0050] a) subjecting the powdery blend obtained in step (iv) to compression to obtain the tablet, or [0051] b1) subjecting the powdery blend obtained in step (iv) to compaction, [0052] b2) milling the compacted material obtained in step (b1) to obtain granules, [0053] b3) optionally mixing the granules obtained in step (b2) and a pharmaceutical excipient to obtain a mixture, and [0054] b4) subjecting the granules obtained in step (b2) or the mixture obtained in step (b3) to compression to obtain the tablet.

[0055] Hence, the phosphate salt of amifampridine of the present invention can be used in the preparation of a tablet through direct compression or dry-granulation, whereby direct compression (above method (a)) is preferred. The phosphate salt of amifampridine of the present invention can be used for the treatment of botulism, myasthenia gravis, congenital myasthenic syndromes (CMS), Lambert-Eaton myasthenic syndrome (LEMS) or fatigue related to a neurological pathology, such as multiple sclerosis and amyotrophic lateral sclerosis (ALS). The invention is further illustrated by reference to the following examples.

EXAMPLES

Analytical Methods:

Appearance:

[0056] The appearance of the drug substance is determined by visual inspection.

[0057] .sup.1H-NMR:

[0058] The drug substance (about 20 mg) is weighed, dissolved in 0.6 mL deuterium oxide (D.sub.2O) and measured using Bruker 500 MHz Avance™ II HD NMR spectrometer.

Melting Point:

[0059] The melting point (m.p.) is measured automatically by a digital melting point system Mettler Toledo MP70 according to Ph. Eur 2.2.14.

[0060] DSC:

[0061] DSC data are obtained on Netzsch Phoenix DSC 204 F1. The drug substance sample (about 10 mg) is placed into a DSC pan and weight accurately recorded. Scaled aluminum pan with one pinhole is used for analysis. The sample is heated under nitrogen atmosphere at a rate of 10° C./min.

Related Substances by HPLC:

Chemicals:

[0062] Water (H.sub.2O); HPLC gradient grade

[0063] Acetonitrile (ACN); HPLC gradient grade

[0064] Trifluoroacetic acid (TFA)

Chromatographic System:

[0065]

TABLE-US-00003 Column: Phenomenex: Gemini C6-Hexyl, 250 × 4.6 mm, 110 Å Mobile phase: Eluent A: ACN:H.sub.2O (10:90) 0.1% TFA Flowrate: 0.8 mL/min Column 25° C. temperature: Detector: UV 230 nm Inject volume: 20 μL Mode Isocratic Acquisition 15 min time

[0066] Preparation of sample (test solution): The drug substance (about 10 mg) is weighed in a 25 mL volumetric flask, dissolved in acetonitrile and the flask is filled to the mark with acetonitrile.

[0067] XRPD:

[0068] The qualitative PXRD measurements were performed by cGMP-accredited XRPD laboratory DANNALAB B. V. (Enschede, the Netherlands) using XPert MPD system.

[0069] A summary of XRPD method parameters is described below:

[0070] PANalytical X′Pert MPD equipped with LLF x-ray source with Cu-Kα1 radiation, Soller collimators of 0.02, 0.04 rad, divergent slit ½ deg and focusing X-Ray mirror are used. The powder load was 400 mg. The radiation used is Cu-Kα1=1.54059 Å (8 keV); 40 kV-40 mA; measurement range 2-40 degrees; measurement time per scan −2.5 hours; step=0.013° 2θ. During the measurements the sample was rotated at speed 4 s/revolution to improve counting statistic.

Sample Preparation Instruction:

[0071] Prepare the sample using PANalytical sample preparation instruction for loading sample in transmission sample holder. Place the prepared sample holder on the sample changer stage of the XRD instrument and analyze as per the above method at room temperature.

Particle Size Measurement (PSD):

[0072] The particle size determination was done using Malvern Mastersizer 2000 and following dry method using 1 g of the sample:

Typical conditions are:

1. Particle RI: 1.00

[0073] 2. Obscuration range: 1-6%,
3. Air pressure: 2 bar
4. Vibration feed rate: 50%
5. Measurement time: 6 seconds
6. Background time: 6 seconds
7. Measurement: 1 per aliquot

8. No of Aliquots: 1 per SOP

Microscopy:

[0074] Microscopic images were taken by contract laboratory solid-chem GmbH (Bochum, Germany) with the help of a Leica DMRB microscope. The samples were placed, in pure silicone oil, on a microscope slide and examined, under polarizable light, regarding the crystallite size and habit.

Comparative Example

[0075] 50 g amifampridine were dissolved in 1100 ml water by heating the suspension to 80° C. Subsequently, 2 equivalents phosphoric acid (85 wt %) were added. After cooling (ambient temperature) 600 ml water were removed under reduced pressure and 583 ml ethanol were added slowly to the concentrated aqueous solution to initiate the crystallization of the product. The reaction mixture was kept at ambient temperature overnight without stirring. The crystals were isolated by filtration, washed with ethanol and dried under reduced pressure at 60° C. to yield 61 g (64%) of the crystalline form 1 of amifampridine phosphate as a beige, voluminous, cotton-like material (FIG. 6). Related substances by H PLC: unknown impurity-1: 0.178%, unknown Impurity-2; 0.202%, total impurities: 0.38%.

Example 1

[0076] In a 30 L reactor, 300.03 g (2.75 mol) amifampridine were dissolved in 6.0 L ethanol under mechanical stirring at 133 rpm and under heating to T.sub.i=68.4° C. The solution was allowed to cool to T.sub.i=54.3° C. Subsequently, 349.70 g (3.02 mol; 1.1 eq.) phosphoric acid 85% were added within 24 min (T.sub.i,max=60.4° C.). The product precipitated immediately upon addition of the acid, and the resulting suspension was stirred for 17 h 8 min at T.sub.i=22.0° C. Subsequently, the suspension was suction filtered and washed twice with each 1.0 L ethanol and twice with each 1.0 L methyl tert-butyl ether (MTBE). The product was dried under reduced pressure (17 mbar) at T.sub.hath=60° C. to yield 568.08 g (2.74 mol; 99.7%) amifampridine phosphate as a white, free flowing solid (irregular shape, mainly flakes; FIGS. 3 and 4).

[0077] .sup.1H-NMR (D.sub.2O, 500 MHz): δ=7.64 (dd, 1H, .sup.3J.sub.1=6.6 Hz; .sup.3J.sub.2=1.2 Hz), 7.65 (d, 1H, .sup.3J=1.21 Hz), 6.78 (d, 1H, .sup.3J=6.6 Hz).

[0078] m.p.: 225.0-230.4° C.; DSC: onset: 228.06° C., peak: 231.37° C., endset: 236.62° C. XRPD: form 1 (FIG. 1).

[0079] PSD by Malvern (by volume): D.sub.90: 15.028 μm, D.sub.50: 4.396 μm, D.sub.10: 0.474 μm.

[0080] Related substances by HPLC: unknown impurity-1: not detected, unknown impurity-2: not detected, total impurities: not detected.

Example 2

[0081] 7.01 g amifampridine were dissolved in 105 ml methanol at 30° C. under mechanical stirring. The solution was allowed to cool to 19.5° C. Subsequently, phosphoric acid (85 wt %) was added dropwise within 15 min under stirring. A precipitate appears immediately upon addition. After the addition of phosphoric acid, the obtained suspension was stirred for 15 min and subsequently cooled to 2° C. and stirred for 1 h. The precipitate was isolated by filtration and washed with methanol. The product was dried at T.sub.bath=60° C. under reduced pressure (19 mbar), which yields form 1 with about 15% form 2 (FIG. 2) as a white, free-flowing solid (plate-like shape; FIG. 5).