阅读说明：本技术 曲前列环素的特丁胺盐 (Triprostinil tert-butylamine salt ) 是由 D·M·雷米克 于 2019-09-11 设计创作，主要内容包括：本申请公开了曲前列环素的新型盐及其制备方法和用途。(The present application discloses novel salts of treprostinil, methods of preparation and uses thereof.)

1. Treprostinil tert-butylamine salt having the structure:

。

2. the treprostinil tetramine salt of claim 1, in crystalline form characterized by an XRD pattern obtained from a CuK α source (λ = 1.54060 a) comprising a peak at 5.1 ° and a peak at least one of 10.2 °, 20.5 ° and 6.8 ° with a diffraction angle tolerance of 0.2 °.

3. The treprostinil tetramine salt of claim 2, wherein the XRD pattern comprises peaks at each of 10.2 °, 20.5 ° and 6.8 ° with a tolerance of diffraction angles of 0.2 °.

4. The treprostinil tetramine salt of claim 3, wherein the XRD pattern further comprises peaks at least one of 13.7 °, 14.5 °, 16.3 °, 18.7 °, 19.6 ° and 21.5 ° with a tolerance of diffraction angles of 0.2 °.

5. The treprostinil tetramine salt of claim 4, wherein the XRD pattern further comprises peaks at each of 13.7 °, 14.5 °, 16.3 °, 18.7 °, 19.6 ° and 21.5 ° with a tolerance of diffraction angles of 0.2 °.

6. A pharmaceutical composition comprising treprostinil tetramine according to any one of claims 1 to 4 and at least one of a pharmaceutically acceptable carrier, excipient or diluent.

7. The pharmaceutical composition of claim 5, further comprising an additional substance having pharmacological activity.

8. The pharmaceutical composition of claim 6, wherein the additional substance having pharmacological activity is insulin.

9. The pharmaceutical composition of claim 7, wherein the insulin is insulin lispro.

10. A method of treating or preventing hyperglycemia in a subject in need thereof, comprising administering a pharmaceutically effective amount of the pharmaceutical composition of claim 8.

11. A method of treating or preventing hypertension in a subject in need thereof, comprising administering a pharmaceutically effective amount of treprostinil tert-butylamine salt according to any one of claims 1 to 5.

12. Use of treprostinil tert-butylamine salt according to any one of claims 1 to 5 as a reference standard for determining the efficacy of treprostinil in a composition comprising treprostinil.

13. A method of making the treprostinil tert-butylamine salt of any one of claims 1-5, comprising:

a) contacting treprostinil free acid with an anti-solvent to produce a suspension;

b) contacting the suspension with a solution comprising tert-butylamine (tert-butylamine);

c) the resulting solid treprostinil tert-butylamine salt was isolated.

Examples

Preparation of tert-butylamine 2- [ [ (1R,2R,3aS,9aS) -2-hydroxy-1- [ (3S) -3-hydroxyoctyl ] -2,3,3a,4,9,9 a-hexahydro-1H-cyclopenta [ g ] naphthalen-5-yl ] oxy ] acetate (Treprostinil erbumine)

While stirring at room temperature, treprostinil free acid (100 mg) was added to acetone (2 mL). The suspension was heated to 50 ℃. In another vessel, tert-butylamine (26 mg, 1.4 eq) was mixed with acetone (1 mL). The base solution was added dropwise and the suspension became a solution within a few minutes, after which time a suspension was formed. Acetone (1 mL) was added and mixing was continued for 2 hours. The mixture was stirred and cooled overnight. The white solid was isolated by vacuum filtration on Whatman paper. The resulting white solid cake was air dried in situ to give 99mg (83% yield) of the title compound.

X-ray powder diffraction (XRD) of crystalline treprostinil tert-butylamine

The XRD pattern of the crystalline solid was obtained on a Bruker D4 Endeavor X-ray powder diffractometer equipped with a CuK α source λ = 1.54060 a and a Vantec detector operating at 35 kV and 50 mA. The sample was scanned at 4 to 40 ° 2 θ, step size 0.008 ° 2 θ, scan rate 0.5 sec/step, with 0.6mm divergence slit, 5.28 fixed anti-scatter slits, and 9.5mm detector slits. The dry powder was loaded on a quartz sample holder and a smooth surface was obtained using a glass slide. The diffraction pattern of the crystalline form was collected at ambient temperature and relative humidity. It is well known in the crystallography art that for any given crystalline form, the relative intensities of the diffraction peaks may vary due to preferred orientations resulting from factors such as crystal morphology and habit. Where there is an effect of preferred orientation, the peak intensity changes, but the characteristic peak position of the polymorph does not. See, for example, United States Pharmacopeia #23, National Formulary #18, pages 1843-. Furthermore, it is also well known in the crystallography art that the angular peak positions may vary slightly for any given crystalline form. For example, the peak positions may shift due to changes in temperature or humidity, sample displacement, or the presence or absence of an internal standard when analyzing a sample. In the present case, peak position variability of ± 0.2 in 2 θ would take these potential variations into account without affecting the unambiguous identification of the indicated crystalline form. The crystalline form can be determined based on any unique combination of distinct peaks (in ° 2 θ), typically the more prominent peaks. The crystal form diffraction patterns collected at ambient temperature and relative humidity were adjusted based on NIST 675 standard peaks at 8.853 and 26.774 ° 2 Θ.

As described above, a sample prepared of crystalline tert-butylamine salt was analysed by XRD which is characterised by an XRD pattern having diffraction peaks as described in table 1 below, in particular having a peak at 5.1 ° in combination with one or more peaks selected from 10.2 °, 20.5 ° and 6.8 ° with a diffraction angle tolerance of 0.2 °.

Table 1: XRD peaks of crystalline treprostinil tert-butylamine

Peak(s)	Angle (° 2-theta) +/-0.2 °	Relative intensity (% of the strongest peak)
			1	5.1	100
2	6.8	36.8
			3	10.2	55.7
4	13.7	34.3
			5	14.5	18.1
6	16.3	17.2
			7	18.7	15.2
8	19.6	18.8
			9	20.5	42.8
10	21.5	17.2

Thermal characterization of treprostinil tert-butylamine salt and free acid

Samples of treprostinil tertamide salt prepared as described above and treprostinil free acid purchased from chemical supply were analyzed for thermal stability by thermogravimetric analysis (TGA) on a TA Instruments TGA-Q5000 thermogravimetric analyzer and Differential Scanning Calorimetry (DSC) on a TA Instruments Q2000 differential scanning calorimeter.

FIG. 1 (the tert-butylamine salt) and FIG. 2 (the free acid) show a superposition of the TGA thermogram at 25-225 ℃ and the DSC thermogram at 25-300 ℃. The TGA data shows a weight loss of 0.2318% between 25 and 100 ℃ for the tetrabutylammonium salt and 1.090% between 26 and 70 ℃ for the free acid. The DSC data for the tert-butylamine salt shows a single endothermic event (possibly melting or decomposition) starting at 143.73 ℃ and the DSC data for the free acid shows three endothermic events (71.17 ℃, 95.57 ℃ and 125.15 ℃) corresponding to possible hydrates and at least two anhydrous crystalline forms. Thermal characterization data indicates that the salt of tert-butylamine has improved thermal stability relative to the free acid and is thermally stable up to at least 100 ℃.

Hygroscopic properties of treprostinil tertamide and sodium salts

Hygroscopicity analyses of treprostinil tetramine prepared as described above and treprostinil sodium purchased from chemical supply were performed on a TA Instruments Q5000SA adsorption analyzer. The relative humidity was increased by 5% up to 95% and then decreased at 5% intervals back to 5% relative humidity on the dried sample at 25 ℃ to generate the moisture absorption curve. The sample was equilibrated at each increment until the weight percent change <0.0100 for 5 minutes.

Fig. 3 (tert-butylamine salt) and 4 (sodium salt) provide dynamic vapor adsorption/reabsorption isotherms. For the sodium salt, at the 80% relative humidity point, the sample had a 25% weight gain, which was classified as very hygroscopic, and once adsorbed, desorption of water did not appear to begin until the relative humidity dropped to 45% RH. XRD analysis was performed on samples collected at various points during adsorption/desorption as described above and the data showed that when the sodium salt was exposed to high relative humidity, the crystal form changed, followed by an irreversible change to an amorphous or poorly crystalline state when returned to low relative humidity. On the other hand, it was surprising that for the salts of terbutaline, at the 80% relative humidity point, the weight gain of the sample was < 0.2%, characterized as non-hygroscopic to slightly hygroscopic (as described in the European Pharmacopoeia on line 9th edition, monograph 5.11 (European Pharmacopoeia on 9th edition), although the experimental conditions were different from those described therein). XRD analysis was performed and showed no change in physical form in amorphization as shown by the sodium salt. These data support the unexpected lack of hygroscopicity of the salts of tert-butylamine, which allows improved storage conditions and efficacy control under ambient conditions. Thus, both the endothermic and hygroscopic data unexpectedly indicate that the salts of tert-butylamine have significantly improved physical stability relative to the free acid and sodium salts.