PHARMACEUTICAL COMPOUND, THE METHOD OF ITS MAKING AND USE AS MEDICINAL AGENT
20220073474 · 2022-03-10
Inventors
Cpc classification
International classification
Abstract
This invention relates to a crystalline form of the Compound (I), wherein the crystalline form displays its strongest reflection, stated as a 2Θ value, at 25±0.2°, in an X-ray powder diffraction pattern. The invention also relates to a method of making this crystalline form, as well as pharmaceutical compositions comprising thereof. Furthermore, the invention relates to methods of using this crystalline form as a medicament and in the treatment of pain.
##STR00001##
Claims
1. A crystalline form of compound (I), ##STR00005## wherein the crystalline form displays its strongest reflection, stated as a 2Q value, at 25±0.2°, in an X-ray powder diffraction pattern.
2. The crystalline form of claim 1, further displaying one or more reflections, stated as a 2Q value, at one or more of 17.8±0.2°, 19.4±0.2°, 20.9±0.2° and 27.5±0.2°, in an X-ray powder diffraction pattern.
3. The crystalline form of claim 1, wherein the X-ray powder diffraction pattern does not display a reflection, stated as a 2Q value, at 15.1±0.2°.
4. The crystalline form of claim 1, having a melting point of 186° C. to 191° C.
5. The crystalline form of claim 1, having a melting point of 187° C. to 190° C.
6. A method of making the crystalline form of claim 1, the method comprising: (i) dissolving compound (I) in dimethylsulfoxide to provide a solution of compound (I) in dimethylsulfoxide; (ii) adding the solution of compound (I) in dimethylsulfoxide to water to provide a suspension of compound (I); (iii) allowing the suspended compound (I) in the suspension of compound (I) to precipitate to provide the crystalline form as defined in claim 1.
7. The method of claim 6, wherein the solution of compound (I) in dimethylsulfoxide is a saturated solution of compound (I) in dimethylsulfoxide.
8. The method of claim 6, wherein adding the solution of compound (I) in dimethylsulfoxide to water is carried out at a temperature of from 15° C. to 25° C.
9. The method of claim 6, wherein allowing the suspended compound (I) in the suspension of compound (I) to precipitate is carried out at a temperature of from 15° C. to 25° C.
10. A pharmaceutical composition comprising the crystalline form of claim 1, and at least one pharmaceutically acceptable excipient.
11. The pharmaceutical composition of claim 10 comprising one or more further pharmaceutically active agents.
12-16. (canceled)
17. A method of treating or preventing neuropathic pain in a human or animal patient comprising administering to a patient in need thereof an effective amount of a crystalline form of claim 1.
18. A method of treating or preventing nociceptive pain in a human or animal patient comprising administering to a patient in need thereof an effective amount of a crystalline form of claim 1.
19. A method of treating or preventing diabetic neuropathy in a human or animal patient comprising administering to a patient in need thereof an effective amount of a crystalline form of claim 1.
20. A method of treating or preventing neuropathic pain in a human or animal patient comprising administering to a patient in need thereof an effective amount of the pharmaceutical composition according to claim 10.
21. A method of treating or preventing nociceptive pain in a human or animal patient comprising administering to a patient in need thereof an effective amount of the pharmaceutical composition according to claim 10.
22. A method of treating or preventing diabetic neuropathy in a human or animal patient comprising administering to a patient in need thereof an effective amount of the pharmaceutical composition according to claim 10.
Description
[0078] These and other aspects of the invention will now be described with reference to the accompanying Figures, in which:
[0079]
[0080]
[0081]
[0082]
EXPERIMENTAL SECTION
[0083] X-ray powder diffraction studies were carried out using a «Siemens D500» diffractometer (copper radiation, graphite monochromator on the secondary beam, the Bragg-Brentano geometry).
[0084] Monocrystalline X-ray studies were carried out on a diffractometer «Xcalibur3» (Molybdenum radiation, graphite monochromator, CCD «Sapphire3» detector, ω-scanning, 2θ.sub.MaKC=50°) at room temperature (about 20° C.) and at atmospheric pressure (about 101.325 KPa).
[0085] Thermographic study was performed on a differential scanning calorimeter DSC Q2000 “Thermo Sientific”, heating rate 5° C./min heating range 20-210° C.
[0086] Information on the size of the crystals was obtained on a diffractometer Mastersizer 3000 “Malvent”; the substance was introduced into a dispersant containing 1% aqueous sodium lauryl sulfate solution, kept under ultrasound for 5 minutes, laser quenching 8-13%.
[0087] Micronization was carried out on a laboratory of Tube Mill control from IKA, Germany, using methods known in the art.
EXAMPLES
[0088] The following non-limiting examples further illustrate the present invention.
Comparative Example 1
[0089] Compound (I) was prepared as described in Ukrainian patent No. 108246 and WO2018/067102.
[0090] A sample of this compound (I) was analysed using powder X-ray diffraction using Copper radiation on a «Siemens D500» diffractometer (copper radiation, graphite monochromator on the secondary beam, the Bragg-Brentano geometry). The X-ray powder diffraction pattern of this compound (I) is shown in
[0091] A sample of the same compound (Form I) was analysed using differential scanning calorimetry (DSC). The DSC thermogram is shown in
[0092] Monocrystalline X-ray studies were also carried out on Form I of Compound I. The results are shown in Table 1 below. Crystals of Form I were obtained by the long isothermic (20° C.) evaporation (approximately 1 month) of a sample of Compound (I) in Form I (Comparative Example 1) previously dissolved in ethyl alcohol in a sample bottle sealed with a lubricant-free glass ground stopper.
Inventive Example 1
[0093] A sample of Compound (I), prepared as described in Ukrainian patent No. 108246 and WO2018/067102, is dissolved in dimethyl sulfoxide. In particular, 1 g of the sample of Compound (I) is dissolved per 10 ml of dimethyl sulfoxide at room temperature (about 20° C.). The solution obtained is slowly added to water at room temperature (10 ml of the solution per 100 ml of water) under vigorous stirring and the suspension formed is mixed for 30 minutes. The stirring is then stopped and the suspension is left for precipitation for three hours. The dispersed precipitate is vacuum-filtered using filtering paper, washed three times with purified water (100 ml each time) and dried in a drying and heating chamber (universal drying and heating chamber FD-115, available from Binder Gmbh, Germany) at 120° C.
[0094] The resulting crystalline sample was analysed using powder X-ray diffraction using Copper radiation on a «Siemens D500» diffractometer (copper radiation, graphite monochromator on the secondary beam, the Bragg-Brentano geometry). The X-ray powder diffraction pattern of the resulting sample is shown in
[0095] A sample of Form II was analysed using differential scanning calorimetry (DSC). The DSC thermogram is shown in
[0096] Monocrystalline X-ray studies were also carried out on Form II of Compound I. To prepare the sample for monocrystalline X-ray study, Form II crystals were obtained by the long isothermic (20° C.) evaporation (approximately one month) of a sample of Compound (I) in Form II (Inventive Example 1) previously dissolved in isobutyl alcohol in a sample bottle sealed with a lubricant-free glass ground stopper. As shown below in Table 1, Form II crystallizes in the monoclinic space group Cc with the parameters of the crystal lattice a=10.9301 (9) A, b=19.7560 (14) A, c=8.5894 (8) A, β=108.165 (11) and a cell volume of 1762.3 (3) A3.
[0097] A comparison of the main crystallographic data of two polymorphic forms of Compound I is given in Table 1.
Comparison of Inventive Example 1 and Comparative Example 1
[0098] The basic crystallographic data and refinement of structures for the Form I and Form II of Compound I (Monocrystalline data).
TABLE-US-00001 TABLE 1 Sample Form I Form II The empirical formula C.sub.18H.sub.16BrClN.sub.2O.sub.2 M.sub.r, a.m.u. 407.69 Temperature, T, K 293 (2) Wavelenght, λ (MoKα), Å 0.71073 Syngony, spatial group Monoclinic, P 2.sub.1/c Monoclinic, Cc The lattice parameters (Å, °) a = 11.7032 (15), a = 10.9301 (9), b = 21.5185 (17), b = 19.7560 (14), c = 14.509 (2) c = 8.5894 (8), β = 92.867 (12) β = 108.165 (11) Cell volume V, Å.sup.3 3649.3 (7) 1762.3 (3) Quantity of formula units, Z 8 4 Density d.sub.x, g/cm.sup.3 1.484 1.537 Attenuation ratio μ (MoKα), Mm.sup.−1 2.411 2.497 Structural factor, F.sub.000 1648 824 Crystal size, mm 0.40 × 0.12 × 0.01 0.15 × 0.12 × 0.05 The angle interval, ° 2.97 ≤ θ ≤ 26.00 3.66 ≤ θ ≤ 26.00 Index constraint −13 ≤ h ≤ 14, −26 ≤ k ≤ −13 ≤ h ≤ 13, −24 ≤ k ≤ 26, −17 ≤ l ≤ 17 24, −10 ≤ l ≤ 9 Reflections measured/independent 26808/6741 (R.sub.int = 0.202) 6317/2698 (R.sub.int = 0.0971) Coverage, % 93.9 97.4 Transmittion T.sub.max/T.sub.min 0.598/0.128 0.885/0.706 Refinement method Full matrix LSM F.sup.2.sub.hkl Data/parameters in LSM 6741/434 2698/222 Goodness of fit, S 0.980 0.964 Flack parameter — 0.02 (2) R-factor for the observed refl (I > 2σI) R.sub.1 = 0.0796, wR2 = 0.1357 R.sub.1 = 0.0590, wR2 = 0.1096 The R-factor for refl R.sub.1 = 0.2195, wR.sup.2 = 0.1837 R.sub.1 = 0.1137, wR.sup.2 = 0.1391 Δρ.sub.max/Δρ.sub.min, E./Å.sup.3 0.806/−0.364 0.381/−0.386
Example 2
[0099] Determination of the Mean Lethal Dose (LD.sub.50 of polymorphic forms of Compound (I)
[0100] The preliminary assignment of a substance to a certain toxicity class is based on an indicator that quantitatively characterizes the acute toxicity of the compound—the dose of the substance that causes a lethal effect in 50% of the population (LD.sub.50). Determination of this indicator in one of the types of experimental animals allows to extrapolate it for other species and justify the dose limits for the first phase of clinical trials.
[0101] To estimate the mean lethal dose of various polymorphic forms of Compound (I) they were administered (in the form of a stabilized suspension on an isotonic sodium chloride solution) at increasing doses orally (one route of administration expected in clinical use, for example) to experimental animals (mice) and the lethal effect was observed in the supervised experimental groups throughout 14 days after administration. To calculate the average dose, mortality/survival rates of the animals by the end of the observation period were used.
[0102] The LD.sub.50 values for samples of Form I and Form II were found to be 1121±190 mg/kg for Form I and 1475±402 mg/kg for Form II. The two LD.sub.50 values are not statistically significantly different, on the basis of which it can be concluded that in the used doses, the two polymorphic modifications of compound (I) (propoxazepam) have comparable toxicity. Based on the LD.sub.50 values for orally administered different polymorphic forms, the compound can be assigned to the IV toxicity class (low toxicity compounds), regardless of the polymorphic modification.
Example 3
Analgesic Activity of Crystalline Forms of Compound (I) in the Model of Thermal Nociception (“Tail Flick” Test)
[0103] Antinociceptive (analgesic) activity was evaluated on a rat tail “tail-flick” model using a focused light beam on rats. Latency time was determined by tail pulling away from the beam source, expressed in seconds. Animals were randomized with respect to the initial value of the latent period, with index from 4 to 12 seconds. The magnitude of the antinociceptive effect was indicated by the increasing of the length of the latent period (pain sensitivity threshold, PST). The initial indices of the PST and its changes 2 hours after the administration of the test compound were compared. The calculation of ED.sub.50 (the average effective dose of analgesic activity in this model for rats) was carried out using the least squares method.
[0104] Three samples of compound (I) were prepared and used in the ‘tail-flick’ test: [0105] The first sample is prepared using the sample of Comparative Example 1, i.e. Form I, in its natural coarse crystalline state. In particular, the coarse crystalline sample of Comparative Example 1 is dispersed in a saline solution and stabilized with Tween 80). The crystalline structure (Form I) remains unchanged. [0106] The second sample was prepared by micronizing (mechanically grinding) the coarse crystalline sample of Comparative Example 1, in order to prepare a sample of Form I having a smaller average particle size. In particular, the micronized sample is dispersed in a saline solution and stabilized with Tween 80). The crystalline structure (Form I) remains unchanged. [0107] The third sample is a dispersed sample of Inventive Example 1, i.e. compound (I) in crystalline Form II. In particular, a sample of Inventive Example 1 is dispersed in a saline solution and stabilized with Tween 80). The crystalline structure (Form II) remains unchanged. The third sample has a similar average particle size to the second sample for a side-by-side comparison.
[0108] The results are shown in Table 2.
TABLE-US-00002 TABLE 2 Crystalline form of Form I Form I Compound (I) (coarse crystalline) (micronized) Form II Average particle size (90% of particles by (90% of particles by (90% of particles by volume < 530 microns) volume < ~50 microns) volume < ~50 microns) ED.sub.50 1.82 1.06 0.17 (Average effective dose (50%)), mg/kg
[0109] As shown in Table 2, the value of the average effective dose for Form II (0.17 mg/kg) is substantially lower than for coarse crystalline Form I (1.82 mg/kg). The value of the average effective dose for Form II is also substantially lower (0.17 mg/kg) than for micronized Form I (1.06 mg.Math.kg), despite the average particle sizes being substantially the same (90% of particles by volume <˜50 microns).
[0110] These results therefore indicate that, with regard to nociceptive pain, Form II (Inventive Example 1) surprisingly displays a higher bioavailability compared to Form I (Comparative Example 1).
Example 4
Analgesic Activity of Crystalline Forms of Compound (I) in the Model of Acetic Acid-Induced Pain (Cramps) on Mice
[0111] To reproduce acute visceral and somatic deep pain (nociceptive pain), the method of “acetic acid-induced pain (cramps)” was used in mice. The mice received a 0.75% solution of acetic acid intraperitoneally (at a dose of 0.1 ml/10 g of body weight), 2 hours after the administration of the test compounds. The number of cramps was counted from the first to the 20.sup.th minute (inclusive) after modeling the pathological state. The calculation of ED.sub.50 (the average effective dose of analgesic activity in this model for mice) was carried out using the least squares method.
[0112] Two samples of compound (I) were prepared and used in the ‘tail-flick’ test: [0113] The first sample is a dispersed sample of Form I. This was prepared by adding a hot (65° C.) saturated solution of Comparative Example 1 (Form I) in ethyl alcohol to water at room temperature (about 20° C.) and atmospheric pressure (about 101.325 KPa). Dispersed Form I has a particle size (D(50)) of around 29 μm. [0114] The second sample is a dispersed sample of Inventive Example 1, i.e. compound (I) in crystalline Form II. This sample is prepared by was dispersing Inventive Example 1 in a saline solution stabilized with Tween-80. The crystalline structure remains unchanged. Dispersed Form II has a particle size (D(50)) of around 24.3 μm.
[0115] The results are shown in Table 3.
TABLE-US-00003 TABLE 3 Crystalline form of Compound (I) Dispersed Form I Dispersed Form II ED.sub.50 1.87 mg/kg 0.89 mg/kg (Average effective dose (50%)), mg/kg
[0116] As shown in Table 3, the value of the average effective dose for dispersed Form II (0.89 mg/kg) is substantially lower than for dispersed Form I (1.87 mg/kg). With regard to this pain model (i.e. treatment of nociceptive pain), Form II therefore surprisingly possesses more than double the activity of Form I. These results therefore indicate that, with regard to nociceptive pain, Form II (Inventive Example 1) surprisingly displays a higher bioavailability compared to Form I (Comparative Example 1).
[0117] Compound (I) is known to exhibit analgesic properties, as shown above and in WO2018/067102, for example. The results in Examples 3 and 4 indicate that Form II of compound (I) will display a similar improved bioavailability with regard to neuropathic (central and peripheral) and diabetic neuropathy.
[0118] The following essential features allow us to obtain such a technical result—develop an easy way of obtaining a crystalline form of the Compound (I) and a pharmaceutical composition for use in the treatment and prevention of neuropathic pain, the prevention of nociceptive pain.