DK162053B - TEMPERATURE-STABLE CRYSTALLINIC SALTS OF 7-OEALFA- (2-AMINO-THIAZOL-4-YL) -ALFA- (Z) -METHOXYIMINOACETAMIDOAA-3-OE (1-ETHYL-1-PYRROLIDINIUM) -METHYLAA-3-METHYLAA-3-CE , PHYSICAL MIXTURES OF THE SALTS AND PROCEDURES FOR THE PREPARATION OF THE SULFURIC ACID ADDITION SALT - Google Patents

Description

in

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The present invention relates to temperature-stable semi-synthetic cephalosporin salts, the preparation of which is not disclosed in the literature, the preparation of such salts and mixtures containing these salts.

Aburaki et al. U.S. Patent No. 4,406,899 discloses 7- [α- (2-5 ami nothi-azol-4-yl) -α- (Z) -methoxyiminoacetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate in zwitterion form, and mentions similar acid addition salts (which are present in zwitterion form in injectable preparations) and demonstrates that the zwitterion form has more broad spectrum activity than ceftazidime and cefotaxime.

However, the Aburaki et al. Cephalosporins mentioned are only stable for a few hours as injectable preparations, and the zwitterion form is even as a dry powder at room temperature and loses 30% or more of its effect in storage at elevated temperatures in just one week. (eg 45eC or more) and therefore require specially insulated packaging 15 and / or cooling, which is a disadvantage of packaging and storage compared to ceftazidime and cefotaxime.

Although Aburaki et al. When mentioning acid addition salts, the patent does not specify how these should be prepared or indicate which of these salts may have good dry powder stability if possible.

Kessler et al. mentions in "Comparison of a New Cephalosporin, BMY 28142, with Other Broad-Spectrum / 3-Lactam Antibiotics", Antimicrobial Agents and Chemotherapy, Vol. 27, No. 2, pp. 207-216, Feb. 1985, the sulfate salt, but nothing about how it should be prepared or that this salt has room temperature stability and good powder stability at elevated temperatures.

It has now surprisingly been found that certain crystalline acid addition salts of 7- [α- (2-aminothiazole-4-yl) -α- (Z) -methoxyimino-acetamido) -3 - [(1-methyl-1 pyrrolidinium) methyl] -3-cephem-4-carboxylate in dry form has excellent stability at room temperature and superior stability at -30 at higher temperature compared to the zwitterionic form. By the term "dry powder form" as used herein is meant a moisture content of less than 5% by weight.

These acid addition salts are the crystalline salts of 7- [or- (2-aminothiazol-4-yl) -α- (Z) -methoxyiminoacetamido-3 - [(1-methyl-1-pyrrolidinium) methyl] - 3-cephem-4-carboxylate selected from sulfuric acid, di-nitric acid, mono-hydrochloric acid and di-hydrochloric acid addition salts as well as ortho-phosphoric acid addition salts (1.5-2 moles of orthophosphoric acid per mole of salt, ie a scale from sesqui to di-orthophosphoric acid salts) or soluates

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2 thereof. By the term "crystalline" as used herein is meant at least one form of characteristic molecular order. While the sulfuric acid, di-nitric acid, di-hydrochloric acid and orthophosphoric acid addition salts are prepared in a clearly crystalline form (as confirmed by double refraction under a polarization microscope) with the exact molecular order, the mono-hydrochloric acid addition salt is prepared only to a somewhat regular order ( which is confirmed by slight refraction under polarization microscope) and not with accurately predictable molecular order and thus is "weak" crystalline. As used herein, the term "crystalline" includes not only clearly crystalline salts, but also the "weakly" crystalline mono-saline acid addition to ten salts. ------

The acid addition salts of the invention, when converted into injectable compositions, provide the zwitterion in solution. The Zwitter-15 ion has the structural formula N0-CH3 COO® c'hj 20 The broad-spectrum utility to various organisms of the zwitterion form and thus of aqueous compositions prepared from the salts of this invention is shown by data by Aburaki et al. in U.S. Patent No. 4,406,899.

Aqueous compositions prepared with the acid addition salts of this invention simply by adding sterile water provide acidic solutions which cause unacceptable irritation in rabbit intravenous administration and unacceptable pain in rabbit intramuscular administration. The sulfuric and di-nitric acid addition salts have a reduced solubility which is insufficient for typical injectable preparations. According to the present invention, these unfortunate characteristics have been overcome by using the present salts in physical admixture (i.e., as a mixture of solids) with a pharmaceutically acceptable non-toxic organic or inorganic base in such a ratio that a pH value of ca. 3.5 to approx. 7 by diluting 35 with water to a zwitterion activity of from 1 mg / ml to 400 mg / ml, usually 250 mg / ml (determined by high performance liquid chromatography, hereinafter referred to as HPLC).

A preferred salt of the invention is the crystalline sulfur.

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3 acid addition salts. It is preferred * because its low solubility in water (25 mg / ml) enables high yield of aqueous medium by crystallization.

The crystalline sulfuric acid addition salt is readily prepared by a process comprising: (a) preparing an aqueous mixture of (i) at least 1 mole equivalent of sulfuric acid and (II) zwitterion in an amount above (B) induces crystallization of the sulfuric acid salt, and (c) isolates the crystalline sulfuric acid salt.

The crystalline salts of the invention (hereinafter referred to simply as salts) possess excellent room temperature stability and exhibit a loss of strength (determined by HPLC) of less than 1% when stored for one month at room temperature. These salts also have excellent stability at elevated temperatures and exhibit a strength loss (determined by HPLC) of less than 15% when stored for 1 month at 45-56 ° C.

The sulfuric acid addition salt is a preferred salt of the invention.

It exhibits a strength loss of less than 10% when stored for 1 month at 45-56 ° C. Of great importance is that it has low solubility in water, ie. ca. 25 mg / ml and therefore crystallized from water with minimal residual loss.

The di-nitric acid addition salt of the invention also has a low solubility in water, i. ca. 60 mg / ml, and therefore also gives low residual loss when crystallized from water.

The mono-hydrochloric, di-hydrochloric and sesqui or di-orthophosphoric acid addition salts have a water solubility exceeding 200 mg / ml and are therefore preferably crystallized from organic solvents rather than from water to obtain good yields.

The preparation of the salts according to the invention is described below.

As previously stated, the sulfuric acid addition salt of the invention is prepared by a process comprising the steps of (a) preparing an aqueous mixture of (i) at least one mole equivalent of sulfuric acid and (ii) zwitterion corresponding to said salt in an amount such that it is present in the mixture at a concentration greater than 25 mg / ml, (b) inducing crystallization and (c) isolating the crystalline sulfuric acid addition salt.

In step (a), the zwitterion is preferably used in an amount such that it is present in the mixture at a concentration of about 10%. 100 mg / ml to approx.

200 mg / ml and step (b) is carried out in an aqueous medium without content of

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4 organic solvent. Normally no more than 1. molar equivalents of sulfuric acid is used in step (a). Usually the zwitterion in step (a) is used in an amount such that it is present in the mixture at a concentration less than 500 mg / ml.

Step (a) is readily performed either, by adding the zwitterion in solid form to a sulfuric acid solution (e.g., IN HgSO4) with rapid stirring to form a solution. Alternatively, step (a) can be carried out by dissolving solid zwitterion in water and slowly adding sulfuric acid with stirring to form a solution.

Step (b) is carried out by inducing crystallization, preferably by grafting, and then slurry, preferably for 15 minutes to 2 hours.

It is preferred that this crystallization step be carried out in aqueous medium without organic solvent content, in which case a purity above 98% is usually achieved. Although the presence of organic solvent such as acetone promotes crystallization and enhances the yield by decreasing the solubility of the sulfuric acid addition salt formed in the crystallization medium, it can also promote the precipitation of impurities with a reduced purity. When the zwitterion is used in step (a) in an amount such that it can be present in the mixture in an amount less than 25 mg / ml, organic solvent, preferably acetone, should be included in the crystallization medium in order to give a fair dividend. In case acetone is used, an amount of 0.5 to 10 volumes of acetone per day is suitably used. volume of aqueous crystallization medium.

In step (c), the crystals are separated from the crystallization medium, for stepwise by vacuum filtration, then washed e.g. with acetone / water followed by acetone alone or 0.1N sulfuric acid (eg 1/10 volume) followed by acetone (eg 1/4 volume) and then dried e.g. by vacuum drying at 30-50 ° C for 4-20 hours.

The process of the invention for forming the sulfuric acid salt causes purification of the zwitterionic form due to the limited solubility of the sulfuric acid salt in comparison with the zwitterionic form and can be used to purify the zwitterion without isolating it as a solid. If it is desirable to obtain substantially pure zwitterion (free base) from the sulfuric acid addition salt formed, this can be done by dissolving the salt in water with the addition of Ba (QH) 2 · 8H 2 O in an amount of 90-100% of theoretical at a pH of less than 6.5 to precipitate BaSO4, filter to remove BaSO4, and recover

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Dissolve the filtrate containing the zwitterion dissolved therein and use it as a solution or isolate the solid zwitterion (free base) by lyophilization or by adding acetone to precipitate amorphous zwitterion followed by isolation of the solid zwitterion by vacuum filtration, washing eg. with acetone and vacuum drying. Alternatively, the sulfuric acid addition salt is converted to the free base using ion exchange resin, e.g. "Dowex WGR" (a weakly basic anion exchange resin) and "Dowex XU-40090.01" (a highly acidic cation exchange resin) and then lyophilized.

In view of the preparation of crystalline di-nitric acid addition salt according to the invention, this is achieved by mixing (i) at least two molar equivalents of nitric acid and (ii) zwitterion corresponding to the salt in such an amount that it is present in the mixture at a concentration greater than than 100 mg / ml and then induce crystallization by grafting or rubbing with than a glass rod, diluting with 2-propanol and cooling. The crystalline di-nitric acid addition salt is recovered e.g. by filtration, sequential washing with e.g. 2-propanol-10 ° (50% v / v), 2-propanol and ether and then vacuum drying at 50 ° C for 2 hours.

The mono-hydrochloric acid addition salt of the invention is prepared by dissolving the zwitterion in ca. 1 mol equivalent hydrochloric acid and induce crystallization by adding acetone with stirring and continuing stirring followed by isolation of the crystals, e.g. by vacuum filtration followed by washing with acetone and vacuum drying. Alternatively, the mono-hydrochloric acid addition salt is formed from the di-hydrochloric acid addition salt by slurrying the di-hydrochloric acid addition salt into methylene chloride and adding 1 mole equivalent of triethylamine followed by slurry to form the mono-hydrochloric acid salt which is isolated, e.g. by vacuum filtration followed by washing with methylene chloride and vacuum drying.

The crystalline di-hydrochloric acid addition salt of the invention is prepared by dissolving the zwitterion in at least two molar equivalents of hydrochloric acid, inducing crystallization by the addition of acetone, isolating the crystals, e.g. by vacuum filtration, wash with acetone and vacuum dry.

The crystalline di-orthophosphoric acid addition salt of the invention is prepared by dissolving zwitterion in at least 2 molar equivalents of phosphoric acid, inducing crystallization by the addition of acetone, and isolating the crystals by e.g. vacuum filtration followed by washing first with acetone and then with ether and then vacuum drying. The crystalline sesqui orthophosphoric acid addition salt is formed at the same preparation.

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6 method except that approx. 1 \ mole equivalent of phosphoric acid.

The salts of the invention are converted into injectable compositions by diluting with sterile water and buffered to pH 3.5-7 to achieve an injectable zwitterion concentration of 1 mg / ml to 400 mg / ml zwitterion. Suitable buffering agents include e.g. tri sodium orthophosphate, sodium bicarbonate, sodium citrate, N-methylglucamine, L (+) lysine and L (+) arginine. For intramuscular and intravenous administration to an adult, a total dose of from ca. 750 to approx. 3000 mg daily 10 divided into portions usually sufficient.

The salts of the invention are preferably not converted into injectable compositions simply by adding sterile water because the sulfuric acid and di-nitric acid addition salts are not sufficiently soluble to form normal concentration preparations for administration and because the salts in question dissolve low pH (1.8-2.5), which is painful when injected.

As stated above, it has now been shown that these deficiencies can be overcome by converting the salts into a physical, ie. solid blend with pharmaceutically acceptable, normally solid, non-toxic organic or inorganic bases in such a ratio that a pH is in the range of from 3.5 to approx. 7, preferably from ca. 4 to approx. 6, by diluting the mixture with water to an injectable zwitterion concentration of 1 mg / ml to ~ 400 mg / ml, e.g. a zwitterion activity of 250 mg / ml as determined by the HPLC assay.

The exact ratios of the constituents of the physical blend vary from batch salt to batch salt, since the purity of the salt varies from batch to batch. The ratios of the constituents are determined for a particular batch by a prior titration of a sample to obtain a selected p-H value in the aforementioned range.

The physical mixture is readily stored and shipped in solid form, taking advantage of the stability of the salts, and is readily converted into an injectable preparation simply by, e.g. a nurse or doctor just before use adds water.

The physical mixture is prepared by mixing the salt and base to a uniform mixture, e.g. by using a standard blender in dry atmosphere, and it is then preferably filled in glass or other container, all under aseptic conditions.

The bases for use in the mixture include e.g. trisodium

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7 orthophosphate, sodium bicarbonate, sodium citrate, N-methylglucamine, L (+) lysine and L (+) arginine. L (+) lysine and L (+) arginine are preferred, since mixtures with their contents, upon re-dilution, give injectable compositions which are less painful for injection than animals derived from mixtures containing other bases. L (+) lysine is preferably used in such a ratio that a pH of 3.5-6 is obtained by diluting the mixture with water to give a composition with zwitterionic activity of 250 mg / ml (determined by the HPLC assay) .

The salts of the invention and substantially dry physical mixtures containing them may be stored without cooling or insulated packaging and still retain high strength.

In several of the compositions of the invention, the unstable zwitterion is used as the starting material. The preparation thereof is described in Examples 1-3 of US Patent No. 4,406,899. Herein referred to as zwitterion 7 - [(Z) -2-methoxyimino-2- (2-aminothiazol-4-yl) acetamido-3 - [(1-methyl-1-pyrrolidinyl) methyl] -3-cephem-4 carboxyl at].

BRIEF DESCRIPTION OF THE DRAWINGS The invention will now be further illustrated by way of example with reference to the accompanying drawings, in which: FIG. 1 is a graphical representation of the infrared absorption spectrum of the crystalline sulfuric acid addition salt of 7- [α- (2-amino-thiazol-4-yl) -a- (Z) -methoxyiminoacetamidoj-3 - [(1-methyl-1 pyrrolidinium) methyl] -3-cephem-4-carboxylate sulfate salt in KBr dilution; 2 is a graphical representation of the infrared absorption spectrum of the crystalline sesame phosphoric acid addition salt of 7- [α- (2-aminothi azol-4-yl) -x- (Z) -methoxy-iminoacetamido] -3 - [(1- methyl-1-pyrrolidinium j-methyl J-3-cephem-4-carboxylate in KBr dilution, Figure 3 is a graphical representation of the infrared absorption spectrum of the crystalline diphosphoric acid addition salt of 7- [α- (2-amino-30thi) azol-4-yl) -x- (Z) -methoxy-iminoacetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] -3-cephem-4-carboxylate in KBr dilution.

Example 1

Preparation of the Sulfuric Acid Addition Salt 1.5 g of zwitterion is slowly added to 10 ml of rapidly stirred IN H2 H2 $ 04 (1.59 molar equivalents) at 20-26 ° C. A solution is obtained. Crystallization is then induced by grafting with crystalline sulfuric acid addition salt and the crystalline mass is suspended for ¼ hour. The crystals are separated

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Then, by vacuum filtration, wash with 3 ml of 50% acetone / water (v / v) and with two 5 ml portions of acetone and then vacuum dry at 40-50 ° C overnight.

A typical yield is 1.3 g of sulfuric acid addition salt.

5 Analysis for C

Calculated:% C, 39.44; % H, 4.53; % N, 14,524; % S, 16.62; % H2 O, none Found:% C, 38.91; % H, 4.57; % N, 14.64; % S, 16.71; % H2 O, 1.42

Example 2 Preparation of the Sulfuric Acid Salt 1.5 g of zwitterion is dissolved in 5 ml of water. 5 ml of 1M H2SO4 is slowly added to this solution with stirring. Crystallization is then induced by grafting with crystalline acid addition salt and the crystalline mass is slurried for% hour. The crystals are then separated by vacuum filtration, washed with 3 ml of 50% acetone / water (v / v) and with two 5 ml portions of acetone and vacuum dried at 40-50 ° C overnight.

A typical yield is 1.3 g of sulfuric acid addition salt.

Example 3 Preparation of the (HNO 3) Acid addition salt 300 mg of zwitterion is dissolved in 2N nitric acid (0.5 ml). Rub with a glass rod and dilute the solution with 2-propanol (0.4 ml) and cool. is collected and washed sequentially with 0.4 ml of 2-propanol / H 2 O (1: 1), 2-propanol and then with ether to give 127 mg of dinitrate salt.

Analysis for CjgH ^NgOgS ^HNOj:

Calculated:% C, 37.62; % H, 4.32; % N, 18.47; % S, 10.57;

Found:% C, 36.92; % H, 4.10; % N, 18.08; % S, 10.67; (H2 O content 0.90%) 30

Example 4

Preparation of the monohydric acid salt of 1 g of zwitterion is dissolved in 2.08 ml of 1N HCl (1 mole equivalent) at 20-25 ° C. 30 ml of acetone is added with rapid stirring over 15 35 minutes to form crystals. Stirring is continued for 1 hour. The crystals are isolated by vacuum filtration, washed with 10 ml of acetone and vacuum dried at 50 ° C for 2 hours.

A typical yield is 0.9 g of crystalline monohydrochloric acid salt.

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Analysis for CjgHg ^NgO₂Sg'HCl:

Calculated:% C, 41.37; % H, 4.75; % N, 15.2; % S, 11.63; % C1, 12.86.

Found:% C, 39.32; % H, 4.88; % N, 13.95; % S, 11.28; * C1, 12.44; % H2 O, 4.5.

5 (corrected for H 2 O:% C, 41.17;% N, 14.61;% S, 11.82;% C1, 13.03).

Example 5

Preparation of the disaltic acid addition salt and preparation of monosaltic acid addition salt therefrom 350 mg zwitterion are dissolved in 2 ml of 1N HCl. 10 ml of acetone is added to the resulting solution with rapid stirring and over a 5 minute interval to form crystals. Stirring is continued for a further 5 minutes. Then add another 10 ml of acetone and stir 15 for ½ hour. The crystals are removed by vacuum filtration, washed with two 5 ml portions of acetone and vacuum dried at 40-45 ° C for 24 hours.

A typical yield is 300 mg of crystalline di hydrochloric acid addition salt.

Analysis for c19H24N6 ° 5S2 * 2HC3:

Calculated:% C, 41.38; % H, 4.75; % N, 15.2; % $, 11.62; % C1, 12.8.

Found:% C, 40.78; % H, 4.98; % N, 14.7; % S, 11.25; % H2 O, 1.25.

(corrected for H2 O:% C, 41.1;% N, 14.88;% S, 11.39;% C1, 11.94).

1 g of dihydrochloric acid salt salt prepared as above is suspended in 20 ml of methylene chloride at 20-25 ° C in a sealed flask and 0.28 ml of tri-25 ethylamine is added over a 15 minute interval. The crystalline mass is then slurried for 5 hours. The resulting monohydric acid addition salt crystals are then isolated by vacuum filtration, washed with two 5 ml portions of methylene chloride and vacuum dried at 50 ° C for 2 hours. A typical yield is 800 mg.

30

Example 6

Preparation of di-orthophosphoric acid addition salt 1 g zwitterion is dissolved in 3.4 ml 144 mg / ml H 2 PO 2 (2.2 molar equivalents) at 15 ° C. The resulting solution is clarified by filtration in an appropriate manner. Over a period of 10 minutes, 12 ml of acetone is added to the clarified solution with rapid stirring to form crystals. Stirring is continued for 10 minutes. Then, 30 ml of acetone is added over 10 minutes and stirring is continued for a further time

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About 15 minutes. The crystals are collected by vacuum filtration, washed with two 5 ml portions of acetone and two 5 ml portions of ether and dried under high vacuum for 16 hours.

A typical yield for this type of preparation was 1.1 g of crystal -5 linic di-orthophosphoric acid addition salt.

Analysis for c19H24N605S2'2H3PO4:

Calculated:% C, 33.72; % H, 4.47; % N, 12.42;

Found:% C, 33.43; % H, 4.65; % N, 12.02; % H2 O, 1.82.

(corrected for H2 O:% C, 34.0;% N, 12.2) The Sesqui orthophosphoric acid addition salt is formed as above except that 1.5 molar equivalents of H3 PO4 are used instead of 2.2 molar equivalents.

Example 7 Stability at elevated temperatures

Stability at elevated temperatures was determined by storing the compositions in dry containers at temperatures and for the periods set forth below, and loss or increase in strength was determined by HPLC. Increase in strength in% is indicated by a plus sign in front of a number. Loss of strength of 20 less than 10% over a period of 2 to 4 weeks at 45-56 ° C is usually indicative of less than 10% loss of strength over a period of 2-3 years at room temperature.

Percentage loss 25

45eC 56eC 100eC

(Weeks) (Weeks) (days)

Form 1 2461 2 4 | 1 30 ------------ -.............-..............— I ...... .............IN......

Zwitterion 37 51 71 | 57 - - | 100 H2SO4 salt 2.4 - +5 3 + 5 1.4 | 5 - +6 +3 0 - +6 | 0-10 (HNO3) 2 salt 8.8 3.4 0.68 10.3 | 3.7 2.4 - j - HCl salt 4.8 2.3 6.0 6.4 | 6.4 - - j - 35 (HCl) 2 salt 0 7.4 - yes - 7.2 j 12.4 (H3 PO4) 2 salt 0 3.0 1.0 - 12.7 5.0 - j -

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Example 8

Physical Mixture Testing

Physical mixtures were prepared from crystalline sulfuric acid salt with (a) tri sodium orthophosphate, (b) sodium bicarbonate, (c) L (+) lysine 5, and (d) L (+) arginine. The bases were added in such amounts that, by diluting the mixture with water to a zwitterionic activity of 250 mg / ml (determined by HPLC analysis), pH values were obtained as follows: trisodium orthophosphate (to provide pH 6.0); sodium bicarbonate (to provide pH 6.0); L (+) lysine (to provide pH 6.0 of pH 6.0); L (+) arginine (to provide pH 6.0). Injectable preparations were prepared by reconstituting with sterile water for a zwitterionic activity of 250 mg / ml according to the HPLC analysis. There were no solubility issues. The pain of intramuscular injections (100 mg / kg) on rabbits was below acceptable limits. The least painful preparation was that which contained arginine.

Similar results with good solubility and acceptable pain by intramuscular injection are obtained using the other salts in question in physical admixture with the above bases.

FIG. Figure 1 is the infrared absorption spectrum of the crystalline sulfate salt prepared as described above in Example 1 or 2, which spectrum is obtained using a sample disc prepared with potassium bromide.

The X-ray diffraction pattern of the crystalline sulfate salt of 7- [α- (2-aminothiazol-4-yl) -α- (z) -methoxyiminoacetamido] -3 - [(1-methyl “-25-pyrrolidinium) methyl] -3-cephem 4-Carboxylate prepared as described in Example 1 or 2 was determined with a "Rigaku Powder Diffractometer" using a copper antibody x-ray tube and a nickel filter, the sample being placed in a glass dish. The scan rate was 2 degrees / min. in the range of 5 degrees to 40 degrees, and a 30 diagram was mechanically drawn to indicate the maximum diffraction angles. From this, (d) the distances and relative intensities (I / Γ) were calculated. They are listed below.

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12 d distance (A) ° Ι / Γ (%) 9.20 100 6.80 50 5 5.50 28 5.09 22 4.50 38 4.41 44 4.19 63 10 3.78 38 3.64 44 3.39 25 3.31 31 3.15 47 15

Example 9

Preparation of the sesame phosphate salt

The Zwitterion 0.70 g is dissolved under rapid stirring in 2.2 to 2.4 ml of 85% phosphoric acid (2.1 to 2.2 molar equivalents), diluted 20 1:10 (v / v) with water. The solution is clarified by filtration through a membrane filter having a pore size of 0.22-0.45 microns. 5 to 7 parts by volume of methanol (15-20 ml) are added to the filtrate with rapid stirring over 30 to 60 minutes. Crystals are formed during this process and the rapid stirring is continued for 2 hours. The crystalline product is recovered by vacuum filtration. The product is washed on the filter first with 6 to 8 ml of methanol: acetone 1: 1 (v / v) ratio, ensuring that a tightly packed filter cake is preserved, and then with acetone. The product is dried in vacuo at 50 ° C for 2 hours, typically yield 0.7 to 0.75 g.

30.

Interpretation of IR spectrum (see Fig. 2) (IR, KBr disk).

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Top position (cm-1) Functional group

2800-3400 NH, NH3 +, carboxyl OH

1780 / J-1 actam C = 0 5 1680 Carboxyl C = 0 1660 Amide C = 0

1630 C = N, C = C

1550 Amid OH

980, 1040 P04 = 10

Behavior on heating

Under differential scanning calorimetry, an exothermic conversion was detected at 171.8 ° C.

X-ray diffraction pattern The X-ray diffraction pattern of the above-mentioned sesqui-phosphate salt was obtained by a "Rigaku Powder Diffractometer" in the manner described for the sulfate salt above, with the following results.

20 d Ι / Γ 11.04 32 9.2 16 25 7.89 24 7.02 42 6.7 32 5.5 26 4.64 100 30 4.456 53 4.3 58 3.88 26 3.75 89 3.56 21 35 3, 31 26 3.05 16

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Interpretation of NMR spectrum (* H 90 MHz NMR, D 2 O solution) 5 N- ° CH * S ^ ir u / O «Τ η n <10

Chemical shifts (ppm δ in relation to TSP) Description Integral Attributes to 15 2.0-2.4 Multiples 4 14CH2, 14'CH2 3.04 Singles 3 12CH3 3.3-3.6 Multiples 5 2CH, 13CH2, 13'CH ^

3.94 Doublet 1 2CH

4.12 Singlet 3 20CH3

20 4.12 Doublet 1 11CH

4.8 Doublet 1 11CH

5.42 Doublet 1 6CH

5.88 Doublet 1 7CH

7.21 Singlet 1 18CH

25

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Stability

Time_Temperature Loss 5 1 day 100 ° C - 10.9 3 days 70 ° C - 0 7 days 70 ° C - 1.9 1 week 56 ° C - 1.0 2 weeks 56 ° C - 0 10 4 weeks 56 ° C - 0 1 week 45 ° C - 0 2 weeks 45 ° C - 1.4 4 weeks 45 ° C - 0.7 8 weeks 45 ° C - 1.6 15 1 month 37 ° C - 2.5

Elemental analysis (weight percent) 20 Found Dry basis Theoretical (sesame phosphate) C 35.44 36.3 36.4 H 4.66 4.41 4.7 N 12.88 13.2 13.4 25 H2 O 2.29 * - monohydrate = 2.8% H2 O H3 P04 23.06 23.06 23.06 * Karl Fischer method 30

Example 10

Methanol crystallization of sesame phosphate salt 25 g of recrystallized sulfate salt prepared as described above in Examples 1 or 2 was dissolved in 400 ml of 1,1,2-trichloro-2,2,1-trifluoroethane ("Freon TF") and the solution treated with 50 ml Amber! ite LA2® (Amberiite LA-2® is a water-insoluble aliphatic secondary amine which is soluble in organic solvent whose mineral acid salt tea is also soluble in organic solvent) and 50 ml of water. After powerful

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After stirring the mixture for 70 minutes, the phases were separated and the aqueous phase containing the zwitterion was collected followed by treatment thereof with an additional 40 ml of Amberi ite LA-2®, extraction with an additional 310 ml of Freon TF, decolorising carbon treatment and filtration.

5 1/2 of the above-mentioned aqueous solution of the zwitterion, 33.5 ml, was placed in a stirring vessel and treated with 1.65 ml of 85% phosphoric acid. An additional 1.65 ml of 80% phosphoric acid was mixed with 90 ml of methanol and slowly added to the aqueous zwitterion solution over 30 minutes at 25 ° C. A slurry of the desired sesame phosphate salt 10 was formed and stirring was continued at 25 ° C for 1 hour. The product was then collected by suction filtration, washed with 30 ml of absolute ethanol and then with 15 ml of methylene chloride, and dried in vacuo at 45 ° C for 15 hours to give 9.80, 9.7% on the basis of 7- [a- (2) -amino-thiazol-4-yl) -o- (Z) -methoxyiminoacetamido] -3 - [(1-methyl-1-pyrrolidinyl) -methyl-3-cephem-4-carboxylate phosphate salt containing 1.5 and characterized by differential scanning colorimeter curve counting, infrared absorption spectrum and X-ray diffraction pattern described herein.

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Claims (15)

1. Temperature stable crystalline salt of 7- [α- (2-amino-thiazol-4-yl) -α- (Z) -methoxyiminoacetamido] -3 - [(1-methyl-1-pyrrolidinyl) -methyl] - 3-cephem-4-carboxylate selected from the group consisting of sulfuric acid, di-hydrochloric acid, mono-hydrochloric acid and di-hydrochloric acid addition salts as well as ortho-phosphoric acid addition salts containing 1.5-2 molar equivalents of HgPO 2 or solvates thereof. 2. A crystalline salt according to claim 1, characterized in that it is selected from the group consisting of sulfuric, mono-hydrochloric, di-hydrochloric and orthophosphoric acid addition salts or solvates thereof. Physical blend of the salt according to claim 1, characterized in that it contains a pharmaceutically acceptable non-toxic organic or inorganic base at a ratio which provides a pH of approx. 3.5 to approx. 7 by diluting the mixture with water to an injectable concentration. A physical mixture according to claim 3, characterized in that the salt and the base are present in conditions which provide a pH of approx. 4 to approx. 6 by diluting the mixture with water to an injectable concentration. A physical composition according to claim 4, characterized in that the base is L (+) lysine. A physical mixture according to claim 4, characterized in that the base is [_ (+) arginine. A process for preparing sulfuric acid addition salt according to claim 1, characterized in that (a) prepares an aqueous solution of (i) at least 1 mole equivalent of sulfuric acid and (ii) zwitterion corresponding to this salt, (b) produces crystallization of the sulfuric acid salt, provided that when the zwitterion is present in the mixture at a concentration of less than 25 mg / ml, the crystallization is carried out in the presence of an organic solvent and (c) isolates the crystalline sulfuric acid addition salt. Process according to claim 7, characterized in that step (b) is carried out in an aqueous medium containing no organic solvent. 9. A process according to claim 8, characterized in that the zwitterion of step (a) is used in an amount such that it is present in the mixture at a concentration of less than 500 mg / ml. Process according to claim 7, characterized in that the amount of the zwitterion used in step (a) in step (a) is such that the concentration of zwitterion in the mixture exceeds 25 mg / ml. 11. A process according to claim 8, characterized in that the zwitterion is used in step (a) in an amount such that it is present in the mixture in a concentration of about 5%. 100 mg / ml to approx. 200 mg / ml. The crystalline sulfuric acid addition salt of claim 1. Crystalline 7- [α- (2-aminothiazol-4-yl) -a- (Z) -methoxyiminoacetamido] -3 - [(1-methyl-1-pyrrolidinium)] methyl] -3-cephem-4 -carboxylate sulfate salt according to claim 1, characterized in that it exhibits an X-ray diffraction pattern: d distance (A) ° Ι / Γ (%) 9.20 100 6.80 50 15 5.50 28 5.09 22 4, 58 38 4.41 44 4.19 63 20 3.78 38 3.64 44 3.39 25 3.31 31 3.15 47 25 The crystalline orthophosphoric acid addition salt of claim 1 and hydrates thereof. 15. Crystal 1 in nsk 7- [or- (2-aminothiazol-4-yl) -or- (Z) -methoxyimino-acetamido] -3 - [(1-methyl-1-pyrrolidinium) methyl] 3-cephem-4-carboxylate phosphate according to claim 1, characterized in that it exhibits the following X-ray diffraction pattern. 35 DK 162053B d ur 11.04 32 9.2 16 7.89 24 5 7.02 42 6.7 32 5.5 26 4.64 100 4.456 53 10 4.3 58 3.88 26 3.75 89 3 , 56 21 3.31 26 15 3.05 16 20 25