A COMPOSITION COMPOSED OF CEFTRIAXONE SODIUM AND SULBACTUM SODIUM

Abstract

The present invention provides a composition composed of ceftriaxone sodium and sulbactam sodium, a pharmaceutical composition comprising the same and the application thereof. The composition is characterized in that diffraction peaks at specific angles are included in the X-ray powder diffraction analysis spectrum. The pharmaceutical composition according to the present invention have better antibacterial activity and stability compared with known compositions, and are thus very suitable for the treatment of bacterial infections, especially for the treatment of refractory urogenital system infections caused by Neisseria gonorrhoeae which is drug-resistance to a variety of antibiotics (-lactams, tetracyclines, macrolides, fluoroquinolones and aminoglycosides).

Claims

1.-18. (canceled)

19. A composition composed of ceftriaxone sodium and sulbactam sodium, wherein, in an X-ray powder diffraction analysis spectrum of the composition, the following peaks at 20.2 angles are included: 11.2, 14.3, 17.8, 19.3, 21.2, 22.8 and 23.8.

20. The composition according to claim 19, wherein, in the X-ray powder diffraction analysis spectrum of the composition, the following peaks at 20.2 angles are further included: 12.6, 16.7, 18.4, 20.0, 20.4 and 28.0.

21. The composition according to claim 19, wherein, in an X-ray powder diffraction analysis spectrum of the composition, the following interplanar crystal spacings 0.2 are included: 7.9, 6.2, 5.0, 4.6, 4.2, 3.9 and 3.7.

22. The composition according to claim 21, wherein, in the X-ray powder diffraction analysis spectrum of the composition, the following interplanar crystal spacings 0.2 are further included: 7.1, 5.3, 4.8, 4.4, 4.3 and 3.2.

23. The composition according to claim 19, wherein, in an infrared absorption spectrum, the composition has absorption peaks at the following wave numbers 5 cm.sup.1: 3255, 1742, 1604, 1539, 1398, 1302, 1198, 1124, 1032, 897, 804, 600 and 479.

24. The composition according to claim 21, wherein, in an infrared absorption spectrum, the composition has absorption peaks at the following wave numbers 5 cm.sup.1: 3255, 1742, 1604, 1539, 1398, 1302, 1198, 1124, 1032, 897, 804, 600 and 479.

25. The composition according to claim 23, wherein, in the infrared absorption spectrum, the composition further has absorption peaks at the following wave numbers 5 cm.sup.1: 3441, 3116, 2938, 1500 and 1099.

26. The composition according to claim 19, wherein, in a differential scanning calorimetry analysis spectrum, the composition has an exothermic peak at 269.60.5 C.

27. The composition according to claim 19, wherein, the composition is prepared by a process comprising the following steps of: taking ceftriaxone sodium raw material and sulbactam sodium raw material, grinding until the median particle size D.sub.50 is between 25 m-88 m, and mixing well to give the composition.

28. The composition according to claim 26, wherein, the median particle size D.sub.50 is between 25 m-47 m, 38 m-62 m or 58 m-88 m.

29. A pharmaceutical formulation comprising the composition according to claim 19.

30. A pharmaceutical formulation comprising the composition according to claim 27.

31. The pharmaceutical formulation according to claim 29, wherein, the pharmaceutical formulation is a powder for injection or an injection solution.

32. The pharmaceutical formulation according to claim 31, which is characterized in that the composition is a powder for aseptic powder needle for injection of lyophilized powder for injection.

33. The pharmaceutical formulation according to claim 29, which is characterized in that, in the pharmaceutical composition, the mass ratio of ceftriaxone sodium calculated by ceftriaxone to sulbactam sodium calculated by sulbactam is 1:1 to 8:1.

34. The pharmaceutical formulation according to claim 33, which is characterized in that, the mass ratio is 1:1 to 4:1.

35. A method for treatment of a bacterial infection disease which comprises administering a pharmaceutical formulation according to claim 29.

36. Method according to claim 35, which is characterized in that, the bacteria infection is by the bacterium Neisseria gonorrhoeae having drug-resistance.

37. The method according to claim 36, which is characterized in that, said drug-resistance refers to drug-resistance to a -lactams antibacterial drug, a tetracyclines antibacterial drug, a macrolides antibacterial drug, a fluoroquinolones antibacterial drug or an aminoglycosides antibacterial drug.

38. The method according to claim 35, wherein, the bacterial infection disease is a urogenital system infection disease.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0061] FIG. 1 shows the X-ray powder diffraction analysis spectrum for Lyophilized Powder for Injection A in Comparative Example 1.

[0062] FIG. 2 shows the X-ray powder diffraction analysis spectrum for mixed powder in Comparative Example 2.

[0063] FIG. 3 shows the X-ray powder diffraction analysis spectrum for Composition I in Example 1.

[0064] FIG. 4 shows the infrared absorption analysis spectrum for Composition I in Example 1.

[0065] FIG. 5 shows the X-ray powder diffraction analysis spectrum for Composition II in Example 2.

[0066] FIG. 6 shows the infrared absorption analysis spectrum for Composition II in Example 2.

[0067] FIG. 7 shows the X-ray powder diffraction analysis spectrum for Composition III in Example 3.

[0068] FIG. 8 shows the infrared absorption analysis spectrum for Composition III in Example 3.

EMBODIMENT

[0069] Hereinafter, Comparative Examples, Examples and Test Examples are used to further explain the present invention, but are not intended to limit the technical solutions and the technical effects of the present invention.

[0070] Description: in the following Comparative Examples, Examples and Test Examples:

[0071] (1) The amount of ceftriaxone sodium/ceftriaxone sodium crystals is calculated by ceftriaxone; the amount of sulbactam sodium/sulbactam sodium crystals is calculated by sulbactam, for example, take 500 g ceftriaxone sodium means that the amount of ceftriaxone in the taken ceftriaxone sodium is 500 g.

[0072] (2) The crystal morphology is measured by X-ray powder diffraction method using BRUKER D8 ADVANCE X-ray powder diffraction instrument under the following measurement conditions: CuKa radiation, tube voltage 40 kV, scanning range 5-60, scanning speed 17.7 s/step, step-length 0.02.

[0073] (3) In the infrared absorption spectrum analysis, the samples are pressed into tablets with KBr and then measured;

[0074] (4) STA409 integrated thermal analyzer is from a Germany company NETZSCH and is used in the differential scanning calorimetry method to perform measurement. Measurement conditions: heating rate 10 K/min, flow rate of protective gas N.sub.2 30 mL/min, flow rate of purging gas N.sub.2 20 mL/min, temperature range 35-300.

[0075] (5) The crystallinity is measured using the US Pharmacopoeia USP35-NF30 method.

[0076] (6) The content of impurity is measured using HPLC method according to the European Pharmacopoeia EP8.0 method.

[0077] (7) The clarity of solutions is measured according to the method of Chinese Pharmacopoeia 2015.

Comparative Example 1

[0078] According to the method described in Example 1 of CN102462684A, in a sterile, dark condition and under nitrogen stream, 100 g of ceftriaxone sodium, 50 g of sulbactam sodium and 25 g of mannitol were added into 90 ml of pretreated 70% ethanol solution which was cooled to below 10 which was placed in a non-metallic container, then dissolved completely with stirring; the obtained solution was adjusted to a pH of 6.5 with 0.1 mol/L aqueous solution of sodium bicarbonate, sterile filtration, subpackaged, placed in a lyophilizer filled with inert gas for pre-freeze (48-20), and then subjected to pressure reduction, sublimation and drying to give the lyophilized powder named Lyophilized Powder for Injection A. Sampled and determined for crystal morphology, and the X-ray powder diffraction spectrum was showed in FIG. 1.

Comparative Example 2

[0079] Ceftriaxone sodium was prepared according to the following method described in Example 1 of CN102993215A, comprising: condensing 7-ACT and AE-active ester to give crude product of ceftriaxone sodium; taking 5 g of the crude product and dissolving them by adding 40 ml of water; adding 4.1 g of sodium iso-octoate into the obtained solution at 13 C., stirring until the solution was clear, adjusting the solution to a pH of 7.0 with 10% diluted hydrochloric acid, stirring for additional 30 minutes to give a mother liquor; adding anhydrous ethanol slowly into the mother liquor under stirring and stopping the stirring and the addition when turbidity occurred; after standing for 25 minutes, slowly adding anhydrous ethanol into the solution until a large amount of crystals were formed, all together 150 ml of anhydrous ethanol was used; filtering the resulting crystals, washing the cake to neutral with a mixed solvent of anhydrous ethanol:water=4:1, washing with anhydrous ethanol again and drying at 35 under ambient pressure.

[0080] Sulbactam sodium was prepared according to the following method described in Example 1 of CN103113390A, comprising: taking the crude sulbactam sodium, and adding it into a mixed solvent of N,N-dimethylformamide and water (3:1) the amount of which was 5 times (ml/g) of that of the crude sulbactam sodium, heating to 70; stirring until the crude sulbactam sodium was completely dissolved, keeping the temperature, adjusting the solution to a pH of 5.5, and making the solution flow at a speed of 5 m/s through a direct current magnetic field of 0.5 T, the direction of which was perpendicular to the flow direction of the solution, after magnetic treatment, adding activated carbon into the solution to decoloration, the amount of which was 0.3 times (ml/g) of the mixed solvent, stirring 30 minutes and filtering to give a clear solution; adding ethanol into the clear solution and the amount of which is 5 times (ml/ml)of that of the mixed solvent, filtering to give the cake; washing the cake with distilled water 3 times, and then drying 4 hours under reduced pressure to give sulbactam sodium.

[0081] Taking 1,000 g of ceftriaxone sodium and 500 g of sulbactam sodium prepared according to the above-mentioned method, mixed well by a trough mixer under sterile condition to give mixed powder, which was sampled and subjected to crystal morphology measurement. The X-ray powder diffraction spectrum was showed in FIG. 2. The mixed powder was subpackaged under a sterile condition to give a sterile powder for injection, named sterile powder for injection A.

Comparative Example 3

[0082] Taking saled 2,000 g of ceftriaxone sodium and 1,000 g of sulbactam sodium, mixed well through a trough mixer under sterile condition to give mixed powder, which was sampled and measured for crystal morphology. The X-ray powder diffraction spectrum of the powder was similar to that of FIG. 2. The mixed powder was subpackaged under a sterile condition to give a sterile powder for injection, named sterile powder for injection B.

Example 1

[0083] 1200 g of ceftriaxone sodium and 600 g of sulbactam sodium both prepared according to the method in Comparative Example 2 were taken and grinding together by a ball mill until the particle size D.sub.50 was 25 m-47 m, and then placed in a single-cone ribbon mixer (HF1600 type) and mixed well, with rotating speed of 30 rpm and mixing time was 25 minutes. After the operation was completed, the material was taken out and a crystalline substance named Composition I was obtained.

[0084] The Composition I was sampled and measured for crystal morphology. The X-ray powder diffraction spectrum was showed in FIG. 3. Main data of the spectrum were listed in Table 1.

[0085] Its infrared absorption was measured by infrared absorption spectrum analysis. The spectrum was showed in FIG. 4. The main data of infrared absorption peaks were listed in Table 2.

[0086] The Composition I was analyzed by the differential scanning calorimetry, and it was found that there was an exothermic peak at 269.51.

TABLE-US-00001 TABLE 1 Main Data of X-ray Powder Diffraction Spectrum for Composition I Interplanar 2 Crystal Peak Peak No. Angle () Spacing() Height Aera 1 11.138 7.9372 1502 16530 2 12.521 7.0634 437 4363 3 14.289 6.1934 11014 68918 4 16.638 5.3238 1798 12883 5 17.835 4.9692 1569 11493 6 18.432 4.8096 883 7497 7 19.276 4.6008 2908 21910 8 19.978 4.4406 1034 12897 9 20.453 4.3386 1342 20418 10 21.171 4.1930 1651 17080 11 22.736 3.9079 1889 25822 12 23.769 3.7403 1299 15065 13 27.985 3.1856 2612 25393

TABLE-US-00002 TABLE 2 Main Data of Infrared Absorption Spectrum for Composition I No. Wave Number(cm.sup.1) 1 3441 2 3254 3 3117 4 2937 5 1742 6 1605 7 1539 8 1501 9 1398 10 1302 11 1198 12 1124 13 1099 14 1032 15 897 16 804 17 600 18 480

Example 2

[0087] Taking saled ceftriaxone sodium 500 g and sulbactam sodium 500 g, and grinding together by a ball mill until the particle size D.sub.50 was 38 m-62 m, then placed in a single-cone ribbon mixer (HF1600 type) and mixed well, with rotating speed of 40 rpm and mixing time was 20 minutes. After the operation was completed, the material was taken out and a crystalline substance named Composition II was obtained.

[0088] The Composition II was sampled and measured for crystal morphology. The X-ray powder diffraction spectrum was showed in FIG. 5. Main data of the spectrum were listed in Table 3.

[0089] Its infrared absorption was measured by infrared absorption spectrum analysis. The spectrum was showed in FIG. 6. The main data of infrared absorption peaks were listed in Table 4.

[0090] The Composition II was analyzed by differential scanning calorimetry analysis, and it was found that there was an exothermic peak at 269.67 C.

TABLE-US-00003 TABLE 3 Main Data of X-ray Powder Diffraction for Composition II Interplanar 2 Crystal Peak Peak No. Angle() Spacing() Height Aera 1 11.120 7.9500 1250 15628 2 12.521 7.0635 413 3990 3 14.272 6.2007 8409 51004 4 16.656 5.3181 1282 10862 5 17.815 4.9746 1514 10864 6 18.431 4.8097 733 6124 7 19.292 4.5970 2475 16754 8 19.923 4.4528 537 18373 9 20.453 4.3386 1604 15208 10 21.171 4.1931 1308 13632 11 22.735 3.9080 1626 23393 12 23.825 3.7317 947 14460 13 27.966 3.1878 2090 21313

TABLE-US-00004 TABLE 4 Main Data of Infrared Absorption Spectrum for Composition II No. Wave Number(cm.sup.1) 1 3441 2 3254 3 3115 4 2939 5 1742 6 1605 7 1539 8 1499 9 1398 10 1302 11 1198 12 1124 13 1099 14 1032 15 897 16 804 17 600 18 478

Example 3

[0091] 930 g of commercially available ceftriaxone sodium was taken and ground; 310 g of commercially available sulbactam sodium was taken and ground; and then these two raw materials were ground together by a ball mill until the particle size D.sub.50 was 58 m-88 m. The obtained powder was placed in a single-cone ribbon mixer (HF1600 type) and mixed well, with rotating speed of 25 rpm and mixing time of 10 minutes. After the operation was completed, the material was taken out and a crystalline substance named Composition III was obtained.

[0092] The Composition III was sampled and measured for crystal morphology . The X-ray powder diffraction spectrum was showed in FIG. 7. Main data of the spectrum were listed in Table 5.

[0093] Its infrared absorption was measured by infrared absorption spectrum analysis. The spectrum was showed in FIG. 8. The main data of infrared absorption peaks were listed in Table 6.

[0094] The Composition III was analyzed by differential scanning calorimetry, and it was found that there was an exothermic peak at 269.69 C.

TABLE-US-00005 TABLE 5 Main Data of X-ray Powder Diffraction for Composition III 2 interplanar crystal Peak Peak No. Angle() spacing() Height Aera 1 11.159 7.9227 1519 16847 2 12.576 7.0326 458 4586 3 14.294 6.1913 13132 70185 4 16.658 5.3176 1786 11679 5 17.851 4.9647 1873 10087 6 18.450 4.8049 910 7192 7 19.315 4.5917 3940 24924 8 19.998 4.4362 859 11627 9 20.476 4.3338 1901 13788 10 21.208 4.1858 1446 16328 11 22.701 3.9138 1468 21912 12 23.807 3.7345 1261 15779 13 27.973 3.1870 1935 23623

TABLE-US-00006 TABLE 6 Main Data of Infrared Absorption Spectrum for Composition III No. Wave Number(cm.sup.1) 1 3441 2 3256 3 3117 4 2937 5 1742 6 1603 7 1539 8 1499 9 1398 10 1302 11 1198 12 1124 13 1099 14 1032 15 897 16 804 17 600 18 480

Examples 4-8

[0095] 800 g of ceftriaxone sodium and 200 g of sulbactam sodium; 1,150 g of ceftriaxone sodium and 230 g of sulbactam sodium; 2,700 g of ceftriaxone sodium and 450 g of sulbactam sodium; 1,330 g of ceftriaxone sodium and 190 g of sulbactam sodium; 2,200 g of ceftriaxone sodium and 275 g of sulbactam sodium, all of which were available in the market, were taken respectively and then processed respectively according to the method in Example 2 to get Composition IV-Composition VIII respectively. It was found that the spectra of these five compositions were basically the same as the spectra obtained in Examples 1-3 through X-ray diffraction analysis.

Example 9

[0096] 500 g of ceftriaxone sodium and 500 g of sulbactam sodium both available in the market were taken and ground together by a ball mill until the particle size D.sub.50 was 104 m-175 m. Remaining operations of mixing materials were the same as Example 2. After the operations were completed, the material was taken out and subpackaged under sterile condition to give Sterile Powder for Injection C.

[0097] 500 g of ceftriaxone sodium and 500 g of sulbactam sodium both available in the market were taken again and ground together by a ball mill until the particle size D.sub.50 was below 10 m. Remaining operations of mixing materials were the same as Example 2. After the operations were completed, the material was taken out and subpackaged under a sterile condition to give Sterile Powder for Injection D.

Example 10

[0098] The compositions of Examples 1-8 were taken respectively, and subpackaged under sterile condition to give Sterile Powder for Injection I-Sterile Powder for Injection VIII.

[0099] The compositions of Examples 1-8 were taken respectively and processed according to the conventional process of lyophilized agents to give Lyophilized Powder for Injection I-Lyophilized Powder for Injection VIII.

[0100] The compositions of Examples 1-8 were taken respectively and dissolved with 50 times amount of 0.9% sodium chloride aqueous solution and subpackaged to give Injections I-VIII.

Test Example 1: Stability Test

[0101] Stability test was performed for composition I-Composition VIII, Sterile Powder for Injection I, Sterile Powder for Injection II, Sterile Powder for Injection IV, Lyophilized Powder for Injection I, Lyophilized Powder for Injection II, Sterile Powder for Injection A , Sterile Powder for Injection B, Sterile Powder for Injection C, Sterile Powder for Injection D and Lyophilized Powder for Injection A obtained in the above-mentioned Comparative Examples or Examples.

[0102] Test environment: 30 months of placement in an environment in which the temperature was 25 C.2 C. and the relative humidity was 60%10%.

[0103] Test results: clarity of solution; content of impurities 1, 2, 3 and 4 (wherein, impurities 1, 2 and 3 were respectively impurities A, B and E of ceftriaxone sodium described in EP8.0, and impurity 4 was impurity A of sulbactam sodium described in EP8.0), crystallinity and the like were measured and main test results were shown in Tables 7- 10.

##STR00001##

TABLE-US-00007 TABLE 7 Situations of Various Samples at 0.sup.th Month in Stability Test Con- Con- Con- Con- tent tent tent tent of of of of Impu - Impu - Impu - Impu - Clarity Crys- rity rity rity rity of tallinity 1 2 3 4 Solu- Samples (%) (%) (%) (%) (%) tion Composition I 90.5 0.12 0.08 0.09 0.11 Clear Composition II 92.7 0.15 0.10 0.13 0.12 Clear Composition III 91.2 0.12 0.14 0.15 0.14 Clear Composition IV 92.1 0.11 0.20 0.12 0.10 Clear Composition V 90.4 0.18 0.27 0.19 0.10 Clear Composition VI 91.7 0.09 0.15 0.16 0.18 Clear Composition VII 92.5 0.21 0.10 0.15 0.13 Clear Composition 92.1 0.29 0.27 0.18 0.22 Clear VIII Sterile Powder 90.6 0.14 0.10 0.15 0.15 Clear for Injection I Sterile Powder 92.3 0.14 0.11 0.14 0.14 Clear for Injection II Sterile Powder 90.4 0.17 0.16 0.15 0.12 Clear for Injection IV Lyophilized 87.6 0.12 0.20 0.16 0.17 Clear Powder for Injection I Lyophilized 85.3 0.16 0.20 0.21 0.18 Clear Powder for Injection II Lyophilized / 0.32 0.14 0.22 0.11 Clear Powder for Injection A Sterile Powder / 0.18 0.13 0.26 0.18 Clear for Injection A Sterile Powder / 0.33 0.19 0.18 0.19 Clear for Injection B Sterile Powder 78.2 0.25 0.21 0.20 0.18 Clear for Injection C Sterile Powder 72.1 0.22 0.31 0.23 0.16 Clear for Injection D

TABLE-US-00008 TABLE 8 Situations of Various Samples at 12.sup.th Month in Stability Test Con- Con- Con- Con- tent tent tent tent of of of of Impu - Impu - Impu - Impu - Clarity Crys- rity rity rity rity of tallinity A B E G Solu- Samples (%) (%) (%) (%) (%) tion Composition I 90.4 0.14 0.10 0.11 0.14 Clear Composition II 91.6 0.18 0.11 0.15 0.14 Clear Composition III 91.3 0.19 0.16 0.17 0.17 Clear Composition IV 92.0 0.21 0.25 0.17 0.14 Clear Composition V 89.2 0.16 0.30 0.24 0.13 Clear Composition VI 91.1 0.15 0.21 0.18 0.22 Clear Composition VII 92.4 0.26 0.24 0.21 0.14 Clear Composition 91.9 0.31 0.29 0.19 0.22 Clear VIII Sterile Powder 90.8 0.25 0.14 0.23 0.18 Clear for Injection I Sterile Powder 92.7 0.19 0.17 0.24 0.15 Clear for Injection II Sterile Powder 91.7 0.22 0.19 0.20 0.15 Clear for Injection IV Lyophilized 85.8 0.16 0.23 0.19 0.18 Clear Powder for Injection I Lyophilized 85.7 0.22 0.21 0.26 0.19 Clear Powder for Injection II Lyophilized / 0.53 0.38 0.46 0.54 Clear Powder for Injection A Sterile Powder / 0.69 0.52 0.32 0.41 Clear for Injection A Sterile Powder / 0.53 0.48 0.37 0.43 Clear for Injection B Sterile Powder 75.1 0.34 0.31 0.29 0.34 Clear for Injection C Sterile Powder 73.2 0.29 0.42 0.33 0.30 Clear for Injection D

TABLE-US-00009 TABLE 9 Situations of Various Samples at 24.sup.th Month in Stability Test Con- Con- Con- Con- tent tent tent tent of of of of Impu - Impu - Impu - Impu - Clarity Crys- rity rity rity rity of tallinity A B E G Solu- Samples (%) (%) (%) (%) (%) tion Composition I 89.2 0.18 0.21 0.17 0.25 Clear Composition II 91.0 0.20 0.10 0.19 0.23 Clear Composition III 90.4 0.23 0.20 0.21 0.26 Clear Composition IV 91.7 0.21 0.29 0.23 0.17 Clear Composition V 88.2 0.28 0.33 0.29 0.30 Clear Composition VI 91.0 0.19 0.27 0.28 0.23 Clear Composition VII 91.6 0.33 0.27 0.32 0.22 Clear Composition 91.7 0.35 0.35 0.23 0.29 Clear VIII Sterile Powder 90.1 0.34 0.26 0.37 0.26 Clear for Injection I Sterile Powder 91.9 0.21 0.20 0.35 0.22 Clear for Injection II Sterile Powder 91.4 0.23 0.38 0.24 0.22 Clear for Injection IV Lyophilized 85.2 0.23 0.29 0.33 0.22 Clear Powder for Injection I Lyophilized 84.1 0.30 0.32 0.29 0.30 Clear Powder for Injection II Lyophilized / 0.83 0.77 0.88 0.62 Clear Powder for Injection A Sterile Powder / 0.99 0.82 0.79 0.70 Slightly for Injection A turbid Sterile Powder / 0.78 0.97 0.85 0.75 Slightly for Injection B turbid Sterile Powder 64.0 0.38 0.47 0.59 0.41 Clear for Injection C Sterile Powder 62.2 0.39 0.56 0.44 0.33 Clear for Injection D

TABLE-US-00010 TABLE 10 Situations of Various Samples at 30.sup.th Month in Stability Test Con- Con- Con- Con- tent tent tent tent of of of of Impu - Impu - Impu - Impu - Clarity Crys- rity rity rity rity of tallinity A B E G Solu- Samples (%) (%) (%) (%) (%) tion Composition I 88.1 0.32 0.31 0.33 0.31 Clear Composition II 90.5 0.31 0.29 0.34 0.30 Clear Composition III 88.2 0.38 0.37 0.28 0.34 Clear Composition IV 90.0 0.43 0.32 0.32 0.31 Clear Composition V 87.1 0.37 0.45 0.32 0.33 Clear Composition VI 89.9 0.25 0.33 0.33 0.30 Clear Composition VII 90.5 0.38 0.41 0.39 0.33 Clear Composition 90.2 0.45 0.41 0.31 0.31 Clear VIII Sterile Powder 88.5 0.47 0.45 0.42 0.38 Clear for Injection I Sterile Powder 89.7 0.37 0.39 0.39 0.33 Clear for Injection II Sterile Powder 90.2 0.43 0.42 0.37 0.36 Clear for Injection IV Lyophilized 84.9 0.39 0.39 0.43 0.38 Clear Powder for Injection I Lyophilized 83.7 0.47 0.38 0.42 0.31 Clear Powder for Injection II Lyophilized / 1.23 1.05 1.12 0.78 Slightly Powder for turbid Injection A Sterile Powder / 1.36 1.12 0.94 1.19 Slightly for Injection A turbid Sterile Powder / 1.13 0.99 1.27 1.06 Slightly for Injection B turbid Sterile Powder 60.6 0.57 0.62 0.70 0.47 Clear for Injection C Sterile Powder 53.5 0.65 0.71 0.58 0.42 Clear for Injection D

[0104] It can be seen from the results of the stability test that after 12 months, the compositions and the formulations according to the present invention, as compared with Comparative Examples, had differences in the following aspects such as crystallinity, impurities, clarity of solution and the like, and after 30 months, such differences were more significant.

[0105] The formulations of Comparative Examples did not show a crystalline form and there was no data of crystallinity; however, the compositions and formulations according to the present invention had a crystallinity of up to 90%, which kept substantially unchanged within 30 months, showing very good physical stability.

[0106] At 24.sup.th-30.sup.th month, the formulations of Comparative Examples showed slight turbidity, and the compositions and the formulations according to the present invention did not show turbidity.

[0107] Although, in the formulations of Comparative Examples, the contents of four impurities were equivalent to those in the compositions according to the present invention at 0.sup.th month, the increase of impurities was very significant as time passed, and especially the contents of impurity 1 (an isomer impurity of ceftriaxone) and impurity 4 (an open loop degraded impurity of salbactam) ware greater than 1% and greater than 0.5% respectively at 24.sup.th-30.sup.th month, fail to meet the requirements of EP 8.0; however, the increase of the four impurities in the compositions and the formulations according to the present invention was not significant and fell into a defined scope, indicating very good chemical stability. This finding revealed the possible presence of a close relation between the generation of specific impurities and crystal morphology.

Test Example 2: In Vivo Antibacterial Test

[0108] The stability test samples at 0.sup.th month in test example 1 were sampled and subjected to in vivo antibacterial test; and the stability test samples at 30.sup.th month were sampled again, and subjected to in vivo antibacterial test. Two in vivo antibacterial tests were both performed according to the following method.

2.1 Test Materials

[0109] Test Samples: Compositions I-VIII, Sterile Powder for Injection I, Sterile Powder for Injection II, Sterile Powder for Injection IV, Lyophilized Powder for Injection I, Lyophilized Powder for Injection II, Lyophilized Powder for Injection A, Sterile Powder for Injection A and Sterile Powder for Injection B, all from the test example 1 at 0.sup.th and 30.sup.th month; and control drugs: ceftriaxone sodium, levofloxacin hydrochloride, azithromycin, doxycycline hydrochloride and spectinomycin hydrochloride, wherein, a batch of control drugs were bought at 0.sup.th month of the stability test so as to match with the test samples at 0.sup.th month, and another batch of control drugs were bought at 30.sup.th month of the stability test so as to match with the test samples at 30.sup.th month, and the time when the two batches were bought was within one month from the date of their production.

[0110] Animals: ICR mice weighed 18-22 g, half male and half female.

[0111] Strains: ATCC700603 (Klebsiella pneumoniae), ATCC43069 (Neisseria gonorrhoeae), Q-R (quinolones-resistant Neisseria gonorrhoeae), T-R (tetracyclines-resistant Neisseria gonorrhoeae), PP-R (-lactams-resistant Neisseria gonorrhoeae), MAC-R (macrolides-resistant Neisseria gonorrhoeae), SPE-R (spectinomycin-resistant Neisseria gonorrhoeae) and QS-R (ceftriaxone-resistant Neisseria gonorrhoeae), wherein, ATCC700603, and ATCC43069 were purchased from ATCC; QS-R was prepared according to the method in a reference literature (China Tropical Medicine, 2010, 10 (8), 946-948); and other strains were clinical isolates.

[0112] Protective agent: highly active dry yeast.

2.2 Test Method

[0113] Enhancing Cytotoxicity: the experimental strain was diluted to 10.sup.2 and 10.sup.3 with 5% highly active dry yeast after incubation of 18 hours; 0.5 ml of the strain was injected intraperitoneally into mice to infect them, obtaining a model of septicemia caused by the systemic infection of the experimental mice; no protective measures was provided and thus mice died of infection; aseptic thoracotomy was performed timely on the mice that died of infection, the heart blood or intracavity effusion was taken and evenly coated on corresponding agar plates, and incubated for 18 hours at 37 C.; and after identification, the strain was consistent with the administrated infectious bacteria. The obtained strain was used to infect mice again, and by performing this method 1-2 times, the cytotoxicity could be enhanced and was relatively stable.

[0114] Determining minimum lethal dose (MLD): the experimental strain was diluted with 5% highly active dry yeast to give diluted bacteria solutions at different concentrations: 10.sup.1, 10.sup.2, 10.sup.3 and 10.sup.4, which were injected intraperitoneally into experimental animals (0.5 ml per mouse) respectively to establish a model of septicemia. After infection, the number of dead mice was recorded. The minimum dose causing 100% death of mice was 1 MLD; as a dose in animal protection experiments, a bacterial concentration which was one log lower than 1 MLD and the animals infected by which all survived was named 0.1 MLD. In experiments, a bacteria concentration of 1 MLD was used as the positive control for the infection experiments; and a bacteria concentration of 0.1 MLD was used as the negative control for the infection. In both positive and negative control groups, only the bacteria were administrated and no antibiotic treatment was provided. The mortality of the positive group should be 70% or more, and the mortality of the negative group should be 30% or less.

[0115] Pre-test for Determining the Dose Range of Test Samples: 1 MLD of bacteria was formulated with 5% highly active dry yeast and used to infect mice; immediately and after 6 hours, pre-tests were performed using test samples at three different concentrations (high concentration, medium concentration and low concentration), each dose group 4 mice; the survival number of mice after infection was recorded, and based on this result, the administration dose in the animal protection test was designed. In the first exploration of the administration dose, the fold differences in drug dose concentration of each group may be relatively great, and when the scope was determined, the fold differences may be reduced for another exploration until the suitable drug dose concentration (that is, for the dose group with highest concentration, 70% or more of infected animals survived, and for the lowest dose group, no less than 70% of infected animals died) was found.

[0116] Animal Protection Test:

[0117] After drug dose concentrations at which 100% of mice died and drug dose concentrations at which no death occurred were obtained in the pre-tests for the test strains used to infect mice, mice that were fasted from food other than water for 18 hours before experiment, weighted 18-22 g; half male and half female, were randomly divided into the following groups: (1) ATCC700603 group, (2) ATCC43069 group, (3) Q-R group, (4) T-R group, (5) PP-R group, (6) MAC-R group, (7) SPE-R group and (8) QS-R group; with five test sample concentrations each group, and 10 animals each group. 10 animals were set as blank controls (equal volume of sterile water for injection was administrated). Each mouse was injected intraperitoneally with 0.5 ml of 1 MLD bacteria solution to establish infection models; immediately and after 6 hours, test sample solutions at different concentrations were injected subcutaneously respectively at 0.2 ml per mouse; the mice were observed 7 consecutive days, and the death status of animals in each group were recorded. Autopsy and observation with naked eye were performed on the dead mouse. The 50% effective dose (ED.sub.50) and 95% confidence intervals were calculated by Bliss method; t-test was performed and the significance of difference was compared between groups. The lower the ED.sub.50 was, the better the protection for animals infected by bacteria the test sample provided.

[0118] 2.3 Test Results

[0119] Main results of the animal protection tests were listed in Table 11 and Table 12, wherein, Table 11 showed the in vivo antibacterial ED.sub.50 values of samples at 0.sup.th month of the stability test, and Table 12 showed the in vivo antibacterial ED.sub.50 values of samples at 30.sup.th month of the stability test.

TABLE-US-00011 TABLE 11 In Vivo Antibacterial ED.sub.50 Values of Samples at 0.sup.th Month of the Stability Test (mg/kg) Strains Test ATCC ATCC Q- PP- MAC- SPE- QS Samples 700603 43069 R T-R R R R -R Cef- 35 16 31 46 208 34 52 337 triaxone Sodium Levo- 25 573 48 37 36 67 72 floxacin Hydro- chloride Azithro- 24 43 62 49 487 59 63 mycin Doxy- 37 55 456 52 58 52 105 cycline Hydro- chloride Spectino- 19 40 61 56 38 621 41 mycin Hydro- chloride Com- 21 11 24 34 47 27 42 62 position I Com- 23 13 21 37 50 23 42 65 position II Com- 19 14 22 35 45 25 46 59 position III Com- 22 15 20 32 44 26 47 62 position IV Com- 27 13 26 37 56 27 49 78 position V Com- 30 16 25 38 68 27 48 85 position VI Com- 31 17 25 38 60 29 52 82 position VII Com- 28 15 27 39 66 28 50 84 position VIII Sterile 25 13 24 32 42 25 46 64 Powder for Injection I Sterile 27 10 23 35 48 27 43 67 Powder for Injection II Sterile 24 14 25 34 46 26 44 66 Powder for Injection IV Lyo- 27 14 22 35 43 24 46 62 philized Powder for Injection I Lyo- 28 12 23 37 41 22 45 60 philized Powder for Injection II Lyo- 31 16 27 39 62 26 50 95 philized Powder for Injection A Sterile 32 15 30 40 56 29 48 92 Powder for Injection A Sterile 33 16 32 38 49 24 46 94 Powder for Injection B Note: represents Not Performed.

TABLE-US-00012 TABLE 12 In Vivo Antibacterial ED.sub.50 Values of Samples at 30.sup.th Month of the Stability Test (mg/kg) Strains Test ATCC ATCC PP- MAC- SPE- QS- Samples 700603 43069 Q-R T-R R R R R Cef- 37 21 30 57 224 37 66 373 triaxone Sodium Levo- 30 564 43 42 41 72 82 floxacin Hydro- chloride Azithro- 31 40 59 55 497 65 69 mycin Doxy- 32 51 432 47 62 51 99 cycline Hydro- chloride Spectino- 22 42 68 67 42 571 56 mycin Hydro- chloride Com- 23 16 22 42 50 29 52 71 position I Com- 25 15 20 46 53 27 49 69 position II Com- 19 14 20 45 56 29 48 68 position III Com- 25 17 24 42 51 25 57 70 position IV Com- 31 20 27 48 68 31 55 87 position V Com- 37 20 28 51 75 28 59 88 position VI Com- 38 22 28 47 65 33 57 92 position VII Com- 36 23 26 49 71 31 59 94 position VIII Sterile 26 16 21 41 51 28 48 75 Powder for Injection I Sterile 30 17 20 43 52 28 47 69 Powder for Injection II Sterile 28 16 22 39 56 27 46 71 Powder for Injection IV Lyo- 30 18 26 42 53 25 47 70 philized Powder for Injection I Lyo- 29 17 24 48 61 25 48 67 philized Powder for Injection II Lyo- 35 26 41 78 93 53 79 153 philized Powder for Injection A Sterile 36 28 47 75 102 48 87 174 Powder for Injection A Sterile 35 28 46 77 106 47 91 188 Powder for Injection B Note: represents Not Performed.

[0120] The test results showed that the compositions and the formulations according to the present invention had better antibacterial activity, and especially exhibited significantly stronger inhibitory activity against various drug-resistant Neisseria gonorrhoeae bacteria and in some cases the activity may increase 3-4 times as compared with formulations in Comparative Examples.

[0121] In addition, the compositions and the formulations according to the present invention had relatively good stability of biological activity, and particularly the inhibition against various non-resistant and resistant Neisseria gonorrhoeae may always remain stable; and after 30 months, the biological activity was basically unchanged. After 30 months, the formulations in comparative example showed significant reduction of activity.

[0122] Although the present invention has been described above in detail with general description, specific embodiments and tests, a person skilled in the art may make suitable modifications or improvements thereon based on the present invention. Thus, those modifications or improvements made without departing from the spirits of the present invention all belong to the content of the present invention. The background, summary of the invention, and specific mode for carrying out the present invention are only for illustrative purpose, but not used as evidence for identifying the prior art of the present invention.