Pharmaceutic bacteria-killing preparation

The present invention relates to a pharmaceutical preparation for controlling bacteria.

The inventors of the present invention have developed and disclosed orally effective cephalosporins with a cefem core having a heterocyclic-substituted thioalkylthiol side chain position 3 (see U.S. Patent Application No. 0Ί / Ί29 413, filed July 2, 1991, and Publication No. European Patent Application No. 0 46Ί 64Ί A2). Among these antibiotic compounds, a compound having a thioalkylthiol side chain substituted with a 1,2,3-triazolyl group, i.e. Ί-β - [(Z) -2- (2-amino-4-thiazolyl) -2-hydroxy acid; yiniino (c; tamido] -3- (1,2,3-triazol-4-thio) -thiomethylthio -3-cephem-4-carboxylic acid with the formula shown in the figure (hereinafter referred to as S-1090), which was obtained as a pale yellow powder, has very strong antibacterial activity and is expected to be a promising candidate for the active ingredient antibiotic preparations.

S-1090, however, was not stable enough to give it the form of a medicinal composition. The S-1090 instability is attributed to its properties below.

a) S-1090 generally contains water because of its hygroscopic properties. The problem is that the water content tends to change during the production process, such as powdering, shaping and the like, accompanied by variations in the content of active ingredient in pharmaceutical compositions containing S-1090. This can create a problem with quality control and cause the product to be uncertain in terms of maintaining the quality, content of the active ingredient and the effectiveness of the products.

b) S-1090 produced by the usual method is an amorphous powder and is not sufficiently soluble in water to be recrystallized from it, which creates difficulties in removing impurities found in amorphous S-1090 powder.

c) Even dried S-1090 contains solvent residues that are difficult to remove to a pharmaceutically acceptable level.

The present inventors conducted extensive research to obtain clinically beneficial S-1090 derivatives that were pharmaceutically stable and contained only small or trace amounts of the contaminating solvent and found that S-1090 hydrochloride and its crystalline hydrate have desirable properties such as low toxicity. high water solubility, pharmaceutical stability and strong antibacterial activity.

Thus, the present invention provides a pharmaceutical formulation in which the active ingredient is S-1090 hydrochloride and its crystalline hydrate. The latter exhibits an X-ray diffraction pattern as shown below and in the attached figure 1, and a differential calorimetric scanning curve as shown in the attached figure 2.

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<td> 20</td><td>intensity</td><td> 20</td><td>intensity</td>

<td> 6,24</td><td> 164</td><td> 29,08</td><td> 179</td>

<td> 10,50</td><td> 117</td><td> 29,50</td><td> 104</td>

<td> 10,94</td><td> 1549</td><td> 30,30</td><td> 269</td>

<td> 12,22</td><td> 997</td><td> 30,50</td><td> 405</td>

<td> 12,54</td><td> 687</td><td> 30,74</td><td> 271</td>

<td> 14,10</td><td> 1057</td><td> 31,04</td><td> 74</td>

<td> 16,38</td><td> 209</td><td> 31,80</td><td> 317</td>

<td> 17,90</td><td> 155</td><td> 31,92</td><td> 347</td>

<td> 18,74</td><td> 381</td><td> 32,56</td><td> 58</td>

<td> 462</td><td> 18,94</td><td> 32,98</td><td> 105</td>

<td> 160</td><td> 20,04</td><td> 33,36</td><td> 458</td>

<td> 217</td><td> 20,74</td><td> 33,76</td><td> 231</td>

<td> 2052</td><td> 21,12</td><td> 34,44</td><td> 99</td>

<td> 478</td><td> 21,36</td><td> 35,52</td><td> 119</td>

<td> 315</td><td> 21,68</td><td> 35,80</td><td> 188</td>

<td> 84</td><td> 22,04</td><td> 37,38</td><td> 187</td>

<td> 57</td><td> 22,54</td><td> 37,70</td><td> 127</td>

<td> 1461</td><td> 23,16</td><td> 38,44</td><td> 150</td>

<td> 379</td><td> 23,74</td><td> 39,20</td><td> 133</td>

<td> 432</td><td> 24,32</td><td> 39,96</td><td> 214</td>

<td> 504</td><td> 24,54</td><td> 43,06</td><td> 222</td>

<td> 259</td><td> 25,30</td><td> 44,24</td><td> 104</td>

<td> 25,94</td><td> 521</td><td> 45,02</td><td> 81</td>

<td> 26,14</td><td> 947</td><td> 45,38</td><td> 68</td>

<td> 26,44</td><td> 422</td><td> 45,68</td><td> 94</td>

<td> 27,46</td><td> 362</td><td> 47,20</td><td> 63</td>

<td> 28,02</td><td> 204</td><td> 48,18</td><td> 65</td>

<td> 28,20</td><td> 253</td><td> 55,54</td><td> 67</td>

Measuring conditions: tube; voltage, 40 kV; current, 20 mA; sampling angle, 0.02 °.

In X-ray diffraction data, the value 26 varies depending on the crystal lattice structure, and the peak intensity varies depending on the direction in which the crystal was formed and the crystallization rate. It is well known in science that variation only in intensity does not mean a change in the crystal lattice structure.

Crystalline hydrate of S-1090 hydrochloride stably maintains the attached water, showing a water content corresponding to 1 to 2 hydrates in a wide range of drying conditions. Typically, it maintains stability in the form of crystalline hydrates having a water content equivalent to a degree of hydration from about

1.1 to about 1.3. Although it can be deprived of the attached water and transformed, after drying under low humidity and / or high temperature, into anhydrous crystalline form, if the anhydrous crystals formed are exposed to high humidity, they rapidly absorb water to reach a stable hydrate with a water content of amount corresponding to a degree of hydration from about 1.2 to 1.3, or even 1.8 under certain conditions, even if placed in the atmosphere.

Experts in this field believe that the position in the triazolyl ring in which the triazolyl group, part of the side chain in position 3, binds the sulfur atom is position 4 or 5, depending on the position of the hydrogen atom in the ring and that the proportion of the position of the hydrogen atom binding the ring is affected by a slight variation in conditions and is variable. Thus, for the purposes of the present invention disclosed herein, it includes a pharmaceutical formulation in which S-1090 hydrochloride or its crystalline hydrate is in a form in which the triazolyl substituent binds a sulfur atom at the 4-position or 5-position of the triazolyl ring.

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Figure 1 shows the X-ray diffraction curve of the crystalline hydrate of S-1090 hydrochloride, where the intensity is plotted as the number of counts on the ordinate and the abscissa is 26. The measurement conditions are: tube, Cu; voltage, 40 kV; current, 20 mA; sampling angle, 0.02 °.

Figure 2 shows the differentiation scanning of the calorimetric crystalline hydrate of S-1090 hydrochloride, where the ordinate of the heat is plotted in milliwatts (mW) and the temperature on the abscissa. Figure 2 indicates that heat absorption occurs at a temperature of about 70 ° C - about 120 ° C, with a maximum at about 105 ° C, probably due to the heat of vaporization of the water contained in the crystalline hydrate.

The crystalline hydrate product S-1090 hydrochloride is particularly useful in medicinal applications because it only contains a pharmaceutically acceptable amount of solvent and is low toxic and also stable in quality.

S-1090 hydrochloride or crystalline S-1090 hydrochloride hydrate maintains the attached water under various conditions as shown in the table below, indicating that the S-1090 derivatives are sufficiently stable to give them the pharmaceutical composition and some quality of the final product.

When S-1090 hydrochloride or its crystalline hydrate is administered to a subject, it may be converted to the same active form as the original S-1090 in vivo and exhibits strong antibacterial activity, as demonstrated experimentally in US Application No. 07/729 413. Thus, in vitro tests have shown that S-1090 is effective against Gram-positive bacteria, e.g., Staphylococcus aureus and Streptococcus pyogenes, as well as gram-negative bacteria, e.g., Escherichia coli, Enterobacter cloacar, Pseudomonas aeruginosa. Proteus vulgaris, Proteus mirabilis, Serratia marcescens, Haemophilus influenzae, Klebsiella pneumoniae and Morgania morganii. S-1090 is particularly effective against Gram-negative bacteria when evaluated against Escherichia coli 7437 and Enterobacter cloacae SR233.

The extent of in vivo absorption of S-1090 after administration was assessed by oral administration to mice and measuring blood levels. The results showed high blood levels of S-1090 after oral administration, indicating an excellent degree of absorption.

The pharmaceutical preparation according to the invention contains as active ingredient 7-β - [(Z) -2- (2-amino-4-thiazolyl) -2-hydroxyoxyiminoacetamido] -3- (1,2,3-triazol-4-yl) thiomethylthio hydrochloride -3-cephem-4-carboxylic acid or its crystalline hydrate in an amount of 10 to 66% by weight based on the total weight of the formulation and a pharmaceutically acceptable carrier.

The preparations according to the invention find use in the treatment of bacterial infections in humans and animals or the control of bacteria on unstable products or in disinfectants, which involves the use of an effective amount of S-1090 hydrochloride or its crystalline hydrate.

For oral administration, S-1090 hydrochloride and its crystalline hydrate, together with pharmaceutically acceptable carriers, diluents and excipients, are suitable as standard preparations such as capsules, tablets, granules, powders and suspensions. For parenteral administration, S-1090 hydrochloride and its crystalline hydrate are suitable as solutions or suspensions for subcutaneous, intramuscular, intravenous or intraperitoneal injection. In addition, the preparation of the invention may be formulated as ointments, suppositories, rubbing liquids and the like. A suitable daily dose of S-1090 hydrochloride or its crystalline hydrate may be between about 10 mg and about 4000 mg, preferably 100 mg and about 2000 mg when administered orally, and about 10 mg and about 4000 mg, preferably about 50 mg and about 2000 mg when administered parenterally.

All S-1090 hydrochloride crystalline hydrates exhibit the same X-ray diffraction patterns and differential calorimetric scanning curves as those shown in Figures 1 and 2, respectively, on the basis of the same crystal structure.

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Example I. Stability of S-1090 hydrochloride and its crystalline hydrate

The stability of S-1090 hydrochloride and its crystalline hydrate, as prepared in the examples, was assessed by conducting an accelerated stability test using specific conditions i.e. elevated temperature, humidity or light. Accordingly, the free amine S-1090, S-1090 hydrochloride and the crystalline hydrate of S-1090 hydrochloride were placed under the conditions given in the table below, and titers and color changes were assessed 0.5 and 1 months after the beginning. The results are shown in the table, where the titer is expressed as a percentage (%) of the titer remaining 0.5 and 1 month after the start of the test.

As can be seen from the data in the table, S-1090 hydrochloride or its crystalline hydrate, when compared to the free amine S-1090, show better stability, with respect to the remaining titer (%) and color, indicating that the S-1090 derivatives they have stable pharmaceutical quality, are only contaminated with trace amounts of solvent, and are useful in the preparation of clinically effective preparations. These beneficial characteristics are significant in the medical field, where drug contamination must be avoided.

Table

Accelerated stability test

<td>A sample</td><td>Time months</td><td>50 ° C flooded</td><td>40 ° C humidity 75%</td><td>increased light intensity (10,000 lux)</td>

<td>S-1090</td><td> 0,5</td><td>81.91; zc</td><td>78.75; zc</td><td>87.83; jb</td>

<td>free amine</td><td> 1,0</td><td>67.62; zc</td><td>64.25; zc</td><td>79.98; jb</td>

<td>Powder</td><td> 0,5</td><td>83.22; jb</td><td>95.23; zc</td><td>87.64; zc</td>

<td>hydrochloride</td><td> 1,0</td><td>82.48; jb</td><td>89.86; zc</td><td>84.44; zc</td>

<td>crystalline</td><td> 0,5</td><td>100.00; b.</td><td>100.00; b.</td><td>97.63; b. f.</td>

<td>hydrate</td><td> 1,0</td><td>99.96; b.</td><td>98.38; b.</td><td>95.51; bz</td>

Abbreviations: humidity, humidity; zc, flesh color; jb, light brown; very pale yellow; b., white.

Pharmaceutical preparations

1. Granules Crystalline Hydrate Hydrochloride S-1090 100 mg Lactose 600 mg Corn Starch 290 mg Hydroxypropyl Cellulose 10 mg

The above substances were granulated by a conventional wet method, each 1 g was packaged as a granulated formulation and administered 3 times a day to patients suffering from infections caused by susceptible bacteria.

2. Tablets crystalline hydrate of S-1090 2 hydrochloride (^ 0 mg lactose 65 mg corn starch 32 mg hydroxypropyl cellulose 2 mg magnesium stearate 1 mg

The above substances were granulated by a conventional wet method and were formed into tablets with a diameter of 7.5 mm using a tablet press and administered twice daily to patients suffering from infections caused by susceptible bacteria.

3. Hard capsules crystalline hydrate of S-1090 hydrochloride 50 mg corn starch 47 mig magnesium stearate 1, (5 mg cosmetic talc 1, (5 mg

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The above substances were granulated by a conventional wet method and filled into gelatin capsules size 4 and administered 3 times a day to patients suffering from infections caused by susceptible bacteria.

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Pattern

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NUMBER

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FI6.1

2©

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FIG.2

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