STABLE PHARMACEUTICAL COMPOSITION

Abstract

A stable pharmaceutical composition contains active ingredients of edaravone and dextrocamphol can control the content of unique impurity SCR-756 and impurity SCR-757 thereof.

Claims

1. A pharmaceutical composition, wherein the pharmaceutical composition comprises edaravone, dextrocamphol, sodium metabisulfite and a solvent, and the content of sodium metabisulfite is 0.95-1.05 mg/ml; preferably, the pharmaceutical composition comprises edaravone, dextrocamphol, sodium metabisulfite, a co-solvent and a solvent, and the content of sodium metabisulfite is 0.95-1.05 mg/ml.

2. The pharmaceutical composition according to claim 1, wherein the weight ratio of edaravone to dextrocamphol is 1:1-4:1; preferably, the weight ratio of edaravone to dextrocamphol is 4:1; preferably, the weight ratio of edaravone, dextrocamphol and sodium metabisulphite in the pharmaceutical composition is 4:1:2.

3. The pharmaceutical composition according to claim 1, wherein the content of edaravone in the pharmaceutical composition is 1.0-3.0 mg/ml; preferably, the content of edaravone is 2.0 mg/ml; preferably, the content of dextrocamphol in the pharmaceutical composition is 0.2-1.0 mg/ml, preferably 0.5 mg/ml; preferably, the content of sodium metabisulphite is 0.97-1.03 mg/ml, further preferably, the content of sodium metabisulphite is 1.0 mg/ml; preferably, the co-solvent and the solvent are propylene glycol and water for injection, respectively.

4. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition further contains a compound of formula I or a pharmaceutically acceptable salt thereof, ##STR00016## preferably, the weight ratio of the compound of formula I or a pharmaceutically acceptable salt thereof to edaravone is 0.3% or less.

5. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition further contains a compound of formula II or a pharmaceutically acceptable salt thereof, ##STR00017## preferably, the weight ratio of the compound of formula II or a pharmaceutically acceptable salt thereof to edaravone is 0.3% or less.

6. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition comprises the compound of formula I or a pharmaceutically acceptable salt thereof in a weight ratio to edaravone of 0.3% or less, and the compound of formula II or a pharmaceutically acceptable salt thereof in a weight ratio to edaravone of 0.3% or less.

7. A compound as represented by formula I, or a pharmaceutically acceptable salt thereof, ##STR00018##

8. A compound as represented by formula II, or a pharmaceutically acceptable salt thereof, ##STR00019##

9-10. (canceled)

11. A method for the quality control of the pharmaceutical composition according to claim 1, comprising using the compound as represented by formula I or a pharmaceutically acceptable salt thereof, ##STR00020##

12. A method for the quality control of the pharmaceutical composition according to claim 1, comprising using the compound as represented by formula II or a pharmaceutically acceptable salt thereof, ##STR00021##

13. A method for preventing and/or treating cerebral stroke, comprising administering to a subject to be treated a therapeutically effective amount of the pharmaceutical composition according to claim 1.

14. A method for preventing and/or treating amyotrophic lateral sclerosis or related disorders, comprising administering to a subject to be treated a therapeutically effective amount of the pharmaceutical composition according to claim 1.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0060] FIG. 1 is the COSY spectrum of SCR-756;

[0061] FIG. 2 is the HSQC spectrum of SCR-756;

[0062] FIG. 3 is the HMBC spectrum of SCR-756;

[0063] FIG. 4 is the COSY spectrum of SCR-757;

[0064] FIG. 5 is the HSQC spectrum of SCR-757; and

[0065] FIG. 6 is the HMBC spectrum of SCR-757.

DETAILED DESCRIPTION OF EMBODIMENTS

[0066] The technical solutions of the present invention will be further described in detail below in conjunction with specific examples. It should be understood that the following examples are only to illustrate and explain the present invention, and should not be understood as limiting the scope of protection of the present invention. All techniques achieved based on the above-mentioned content of the present invention fall within the scope of protection of the present invention.

[0067] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

Example 1

Investigation of Amount of Sodium Metabisulfite and Related Substances

[0068] Propylene glycol was heated to 50-60° C., edaravone was added, and the mixture was stirred until complete dissolution to obtain a medicinal solution. The medicinal solution was cooled to 25° C. or less, dextrocamphol (the structure being

##STR00012##

was added, and the mixture was stirred until complete dissolution. Sodium metabisulfite was added to an appropriate amount of water (25° C. or less) and the mixture was stirred until dissolution. The sodium metabisulfite solution was slowly added to the medicinal solution with stirring, and then water was slowly added to make the amount close to the formulation amount. The pH was adjusted to 4.5±0.2 with 0.1 mol/L of hydrochloric acid and sodium hydroxide solution, water was added to achieve the formulation amount, and the resulting medicinal solution was filtered, sealed after nitrogen filling, and sterilized.

[0069] According to the above-mentioned method, pharmaceutical compositions having a sodium metabisulfite content of 0.5 mg/mL, 0.8 mg/mL, 1.0 mg/mL, 1.2 mg/mL, and 1.5 mg/mL were prepared and obtained, and the specific formulations thereof are as shown in Table 1.

TABLE-US-00001 TABLE 1 Formulation information of pharmaceutical compositions F1-F5 Formulation components F1 F2 F3 F4 F5 Edaravone 10 mg 10 mg 10 mg 10 mg 10 mg Dextrocamphol 2.5 mg 2.5 mg 2.5 mg 2.5 mg 2.5 mg Propylene glycol 0.4 mL 0.4 mL 0.4 mL 0.4 mL 0.4 mL (for injection) Sodium metabisulfite 2.5 mg 4 mg 5 mg 6 mg 7.5 mg hydrochloric acid q.s. q.s. q.s. q.s. q.s. Sodium hydroxide q.s. q.s. q.s. q.s. q.s. Water for injection Making up Making up Making up Making up Making up to 5 mL to 5 mL to 5 mL to 5 mL to 5 mL

[0070] The related substances in the pharmaceutical compositions in Table 1 were analyzed. The samples were placed at the temperature of 60° C., and the content of impurity SCR-756, impurity SCR-757 and the total impurities of edaravone in the samples was determined at 0 days and 10 days, respectively. The specific results are as shown in Table 2.

TABLE-US-00002 TABLE 2 Investigation results of content.sup.[1] of impurity SCR-756, impurity SCR-757 and total impurities Sample batch 60° C., 60° C., number Impurity 0 days (%) 10 days (%) F1 SCR-756 0.12 0.39 SCR-757 0.09 0.32 Total impurities 0.48 2.13 F2 SCR-756 0.18 0.47 SCR-757 0.15 0.40 Total impurities 0.66 2.25 F3 SCR-756 0.16 0.29 SCR-757 0.13 0.25 Total impurities 0.57 1.55 F4 SCR-756 0.19 0.39 SCR-757 0.16 0.34 Total impurities 0.64 1.85 F5 SCR-756 0.23 0.38 SCR-757 0.20 0.33 Total impurities 0.77 1.78

[0071] Notes: [1] “Content” refers to the weight ratio of impurity SCR-756, impurity SCR-757 and the total impurities to edaravone in the pharmaceutical composition.

[0072] Analysis method of the related substances: an appropriate amount of the sample was taken and diluted by adding a solvent, i.e., methanol-water (volume ratio: 32:68) to prepare an edaravone solution (about 0.5 mg/mL) as a test solution. An appropriate amount was accurately measured and an edaravone solution (5 μg/mL) was prepared as a control solution by dilution with the solvent. Impurity control SCR-756 and SCR-757 (5 mg each) were accurately weighed and placed in a 100 mL volumetric flask. A solvent was added, and the mixture was sonicated to aid dissolution, diluted to scale, and shaken uniformly to obtain an impurity stock solution. In addition, 10 mg of edaravone control was weighed accurately, placed in a 20 mL volumetric flask, and dissolved by adding 5 mL of methanol. 1 mL of the impurity stock solution was added, and the mixture was diluted to scale by adding a solvent, and shaken uniformly to obtain a system suitability solution. The determination was performed according to the high performance liquid chromatography (General rule 0512, Volume IV, Pharmacopoeia of the People's Republic of China (2015)). Octadecyl silane bonded silica gel was used as the filler (Agilent Eclipse plus C18 4.6×150 mm, 3.5 μm), 0.5% triethylamine (pH 6.3, adjusted with phosphoric acid) was used as mobile phase A, and methanol was used as mobile phase B. Elution with a linear gradient was performed according to the table below. The detection wavelength was 248 nm. 10 μL of the system suitability solution was accurately measured and injected into the liquid chromatograph, and the chromatogram was recorded. The peaks of impurity SCR-756, impurity SCR-757 and edaravone appeared in sequence, and the retention time of edaravone peak was about 6 minutes. The test solution and the control solution (10 μL each) were accurately measured and separately injected into the liquid chromatograph and the chromatograms were record.

TABLE-US-00003 Time (min) Mobile phase A (%) Mobile phase B (%) 0 68 32 8 68 32 15 40 60 30 40 60 31 68 32

Example 2

[0073] Propylene glycol was heated to 50-60° C., edaravone was added, and the mixture was stirred until complete dissolution to obtain a medicinal solution. The medicinal solution was cooled to 25° C. or less, dextrocamphol was added, and the mixture was stirred until complete dissolution. An antioxidant (one or two of sodium metabisulfite, sodium bisulfite, or L-cysteine hydrochloride) was added to an appropriate amount of water at 25° C. or less, and the mixture was stirred until dissolution. The above-mentioned antioxidant solution was slowly added to the medicinal solution with stirring, and then water was slowly added to make the amount close to the formulation amount. The pH was adjusted to 4.5±0.2 with 0.1 mol/L of hydrochloric acid and sodium hydroxide solution, water was added to achieve the formulation amount (as shown in Table 3), and the resulting medicinal solution was filtered, sealed after nitrogen filling, and sterilized. The appearance of the samples and the impurities of edaravone were mainly investigated, and the results are as shown in Table 4.

TABLE-US-00004 TABLE 3 Formu- Formu- Formu- Based on the amount of 3000 vials lation 1 lation 2 lation 3 Edaravone (g) 30 30 30 Dextrocamphol (g) 7.5 7.5 7.5 Sodium metabisulfite (g) 15 15 — Sodium bisulfite (g) — — 15 L-cysteine hydrochloride (g) — 7.5 7.5 Propylene glycol (L) 1.2 1.2 1.2 Water for injection, making up to (L) 15 15 15

TABLE-US-00005 TABLE 4 Edaravone-related Formulation impurities Sample number Appearance (%) Before Formulation 1 Colorless clear 0.29 sterilization liquid Formulation 2 Colorless clear 0.28 liquid Formulation 3 Colorless clear 0.38 liquid After Formulation 1 Colorless clear 0.42 sterilization liquid Formulation 2 Colorless clear 0.49 liquid Formulation 3 Colorless clear 0.53 liquid After Formulation 1 Colorless clear 0.58 sterilization liquid 40° C., Formulation 2 Colorless clear 0.56 1 month liquid Formulation 3 Colorless clear 0.66 liquid

[0074] The amount of edaravone-related impurities in formulations 1 and 2 was significantly lower than that in formulation 3.

[0075] The analysis method of the related substances was as follows:

[0076] the determination was performed according to the high performance liquid chromatography (Appendix VI E, Volume II, Pharmacopoeia of the People's Republic of China (2010)). Octadecyl silane bonded silica gel was used as the filler (Waters sunfire C18, 4.6×250 mm, 5.0 μm), 0.02 mol/L of ammonium acetate solution (pH 4.0, adjusted with glacial acetic acid) was used as mobile phase A, and acetonitrile was used as mobile phase B. Elution with a linear gradient was performed according to the table below.

[0077] Detection wavelength: 254 nm;

[0078] Flow rate: 1.0 ml/min;

[0079] Injection volume: 20 μl;

[0080] Solvent: 0.02 mol/L of ammonium acetate solution (pH 4.0, adjusted with glacial acetic acid)-acetonitrile (volume ratio: 80:20);

[0081] Test solution: 5 ml of the sample was taken, placed in a 20 ml volumetric flask, and diluted to scale by adding the solvent to obtain the test solution;

[0082] Control solution: 1 ml of the test solution was accurately measured, placed in a 200 ml volumetric flask, and diluted to scale by adding the solvent to obtain the control solution.

TABLE-US-00006 Time (min) Mobile phase A (%) Mobile phase B (%) 0 80 20 30 80 20 40 40 60 60 40 60 61 80 20

Example 3

Preparation of Compounds SCR-756 and SCR-757

[0083] ##STR00013##

Synthesis Steps:

(1) Preparation of Intermediate 1

[0084] Na (39.92 g) was added to 1.25 L of anhydrous ethanol in batches while the temperature was kept constant at 25° C.-40° C., and the mixture was stirred at 50° C. until Na was completely dissolved and the solution was clarified. Edaravone (250 g, solid) was added in batches at 0° C.-5° C. in a dark place away from light, and the mixture was stirred to obtain a turbid liquid. The system was protected with N.sub.2 balloon and protected from light with a black plastic bag. Chloroacetone (203.11 g) was dissolved in 750 mL of anhydrous ethanol to prepare an ethanol solution, and the solution was directly added to a dropping funnel. The above-mentioned ethanol solution was added dropwise at −5° C.-0° C. while the system was protected with N.sub.2 balloon and protected from light with a black plastic bag, and the mixture was stirred at 0° C.-5° C. for 2 h. It was monitored by TLC (petroleum ether:ethyl acetate=1:1) that most of the raw material, i.e., edaravone, was consumed. The reaction solution was poured into 4 L of ice water with stirring. The resulting mixture was filtered, and the filter cake was washed with water (600 mL*3), then slurried with ethyl acetate (200 mL), and filtered to obtain intermediate 1 (37 g in total).

(2) Preparation of Intermediate 2

[0085] To a 3 L three-necked flask, 37 g of intermediate 1 was added. Under the protection of N.sub.2 balloon, 650 mL of dichloroethane and 325 mL of anhydrous dioxane were added via an addition funnel. The flask was evacuated and the mixture was stirred until complete dissolution. 4A molecular sieves (100 g) and sulfur trioxide-dioxane adduct (69.41 g) were added in batches successively at 0° C.-5° C. with stirring. The mixture was heated to 100° C., reacted for 8 hours, and then cooled to 25° C. After filtration, the filter cake was rinsed twice with anhydrous dichloromethane, and the filter cake and anhydrous dichloromethane (600 mL) were stirred and filtered to remove the large particles of molecular sieves. The filtrate was stirred for 5 minutes, and then filtered. The filter cake was rinsed twice with anhydrous dichloromethane to obtain a crude product, which was slurried with ethanol and then filtered to obtain intermediate 2 as a solid (30 g).

(3) Preparation of Compounds SCR-756 and SCR-757

[0086] To a 2 L three-neck flask, intermediate 2 (30 g) was added. Under the protection of N.sub.2 balloon, anhydrous MeOH (600 mL) was added via an addition funnel. The flask was evacuated and the mixture was stirred under dry ice bath. The temperature was kept at 0° C.-5° C. NaBH4 (8.85 g) was added in batches. The reaction solution was stirred at 60° C. for 3 h, subjected to rotary evaporation to remove the solvent, and then separated by HPLC (chromatographic column: YMC-Triart Prep C18 250*50 mm*10 μm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; acetonitrile: 0%-28.5%, 19 mins) to obtain compound SCR-756 and compound SCR-757.

[0087] The structures of compound SCR-756 and compound SCR-757 were confirmed by nuclear magnetic resonance.

Nuclear Magnetic Resonance (NMR) Spectroscopy

[0088] Instrument: BRUKER AV-400 nuclear magnetic resonance instrument

[0089] Solvent: DMSO-d.sub.6

[0090] Internal standard: TMS

[0091] Temperature: 300 K.

[0092] .sup.1H NMR data and .sup.13C NMR data of SCR-756 are as shown in Table 5-1 and Table 5-2.

##STR00014##

TABLE-US-00007 TABLE 5-1 Determination results of .sup.1H NMR spectroscopy of SCR-756 Num- Chemical Chemi- ber shifts of cal of related shift pro- Peak protons Assign- (ppm) tons shape (ppm) ment Notes 8.11 3H brs / H.sub.13/H.sub.17/ Active hydrogen H.sub.18 7.69 2H d 7.53, 7.38 H.sub.4/H.sub.6 J = 8.4 Hz 7.53 2H dd 7.69, 7.38 H.sub.1/H.sub.3 J.sub.1 = 7.2 Hz, J.sub.2 = 8.4 Hz 7.38 H dd 7.69, 7.53 H.sub.2 J.sub.1 = J.sub.2 = 7.2 Hz 4.38 H m 3.79, 1.06 H.sub.15 / 3.79 H d 4.38 H.sub.14 J = 6.0 Hz 2.26 3H s / H.sub.12 / 1.06 3H d 4.38 H.sub.16 J = 6.0 Hz

TABLE-US-00008 TABLE 5-2 Determination results of .sup.13C NMR spectroscopy of SCR-756 Chemical Chemical shifts shifts of of long-range Chemical Type related related shift of Assign- protons protons (ppm) carbon ment (ppm) (ppm) 153.1 Quaternary C C.sub.11 / 3.79 148.5 Quaternary C C.sub.9 / 3.79, 2.26 136.3 Quaternary C C.sub.5 / 7.69, 7.53 129.7 Tertiary C C.sub.1, C.sub.3 7.53 7.69, 7.38 127.7 Tertiary C C.sub.2 7.38 7.69 122.7 Tertiary C C.sub.4, C.sub.6 7.69 7.69 96.1 Quaternary C C.sub.10 / 4.38, 3.79, 2.26 67.4 Tertiary C C.sub.15 4.38 3.79, 1.06 63.5 Tertiary C C.sub.14 3.79 4.38, 1.06 19.9 Primary C C.sub.16 1.06 4.38, 3.79 12.4 Primary C C.sub.12 2.26 /

[0093] .sup.1H NMR data and .sup.13C NMR data of SCR-757 are as shown in Table 6-1 and Table 6-2.

##STR00015##

TABLE-US-00009 TABLE 6-1 Determination results of .sup.1H NMR spectroscopy of SCR-757 Num- Chemical Chemi- ber shifts of cal of related shift pro- Peak protons Assign- (ppm) tons shape (ppm) ment Notes 7.87 3H brs / H.sub.13/H.sub.17/ Active hydrogen H.sub.18 7.68 2H d 7.56, 7.42 H.sub.4/H.sub.6 J = 8.4 Hz 7.56 2H dd 7.68, 7.42 H.sub.1/H.sub.3 J.sub.1 = 7.2 Hz, J.sub.2 = 8.4 Hz 7.42 H dd 7.68, 7.56 H.sub.2 J.sub.1 = J.sub.2 = 7.2 Hz 4.86 H m 3.70, 1.15 H.sub.15 / 3.70 H d 4.86 H.sub.14 J = 3.2 Hz 2.26 3H s / H.sub.12 / 1.15 3H d 4.86 H.sub.16 J = 6.4 Hz

TABLE-US-00010 TABLE 6-2 Determination results of .sup.13C NMR spectroscopy of SCR-757 Chemical Chemical shifts shifts of of long-range Chemical related related shift Type of Assign- protons protons (ppm) carbon ment (ppm) (ppm) 154.0 Quaternary C C.sub.11 / 3.70 148.7 Quaternary C C.sub.9 / 3.70, 2.26 135.7 Quaternary C C.sub.5 / 7.68, 7.56 129.9 Tertiary C C.sub.1, C.sub.3 7.56 7.68, 7.42 128.1 Tertiary C C.sub.2 7.42 7.68 122.9 Tertiary C C.sub.4, C.sub.6 7.68 7.42 96.9 Quaternary C C.sub.10 / 4.86, 3.70, 2.26 67.4 Tertiary C C.sub.15 4.86 3.70, 1.15 62.3 Tertiary C C.sub.14 3.70 4.86, 1.15 22.0 Primary C C.sub.16 1.15 3.70 11.9 Primary C C.sub.12 2.26 /

[0094] In addition, the COSY spectrum, HSQC spectrum, and HMBC spectrum of SCR-756 and SCR-757 are as shown in FIG. 1 to FIG. 6, respectively.

[0095] The results show that SCR-756 has the structure as shown in formula I, and SCR-757 has the structure as shown in formula II.

[0096] The embodiments of the present invention are illustrated as above. However, the present invention is not limited to the above-mentioned embodiments. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.