NL8601991A - CEPHALOSPORINE SALTS, THE PREPARATION THEREOF AND INJECTABLE PREPARATIONS THEREOF. - Google Patents

Description

* i i VO 8302

Titlei cephalosporin saltan, its preparation and injectable preparations thereof.

This invention relates to temperature stable semisynthetic cephalosporin salts, the preparation of which has not been described in the literature, to the preparation of such salts and mixtures containing these salts.

In U.S. Patent No. 4,406,899, 7- [ct- (2-amino-thiazol-4-yl) -a- (2) -methoximino-acetamido] -3 - [(1-methyl-1-pyrrolidinio) - methyl] -3-cephem-4-carboxylate in the zwitterion form and corresponding acid addition salts are reported (which are present in the zwitterion form in injectable preparations) and the zwitterion 10 is shown to show a broader spectrum of activity than ceftazidime and cefotaxime.

However, the aforementioned cephalosporins are stable only for several hours as an injectable preparation and the zwitterion form, even as a dry powder, is unstable at room temperature and loses 30% or more of its activity on storage at elevated temperature after 1 week (eg. 45 ° C and above) so that special insulated packaging and / or cooling is required, which is a packaging and storage disadvantage compared to ceftazidime and cefotaxime.

Although the aforementioned US patent mentions acid addition salts, it is not disclosed how to make them and whether one of these salts in dry powder form has good stability. By Kessler et al., "Comparison of a New Cephalosporin, BMY 28142, with Other Broad-Spectrum β-Lactam Antibiotics", Antimicrobial Agents and Chemotherapy, Vol. 27, no. 2, biz. 207-216, February 1985, the sulfate salt is mentioned, but not how to prepare it or whether this salt is stable at room temperature and is well stable in dry powder form at elevated temperature.

It has been discovered that certain crystalline acid addition salts of 7- [a- (2-aminothiazol-4-yl) -a- (Z) -methoxyiminoacetamido] -3 - [(1-methyl-1-pyrrolidinio) -methyl] -3- cefem-4-carboxylate in dry powder form have excellent room temperature stability and have superior elevated temperature stability compared to the zwitterionic form.

The term "dry powder form" as used herein means a moisture content of less than 5% by weight. These acid addition salts are the crystalline salts of 7- [α- (2-aminothiazol-4-yl) -a- (Z) -methoxyiminoacetamido] -S5G-59: 4 i-5 t-2 -3- [(1-methyl -1-pyrrolidinio) -methyl] -3-cephem-4-carboxylate selected from the group consisting of sulfuric acid, di-nitric acid, mono-hydrochloric acid, and di-hydrochloric acid addition salts and orthophosphoric acid addition salts (1.5-2 mol orthophosphoric acid per mole of salt, eg, a range from the sequence to the 5 di-orthophosphoric acid salts) or sulfates thereof.

The term "crystalline" as used herein means at least some characteristic arrangement of the molecules. Although herein the sulfuric, di-nitric, di-hydrochloric and orthophosphoric acid addition salts are prepared in clear crystalline form (as evidenced by birefringence under a polarizing microscope) with an accurate arrangement of molecules, the mono hydrochloric acid addition salt is prepared with only a certain regularity in the arrangement of the molecules (as evidenced by poor birefringence under a polarizing microscope) with an inaccurately predictable arrangement and thus is "poorly" crystalline.

The term "crystalline" is used herein to include not only the clear crystalline salts, but also the "bad" looking crystalline mono-hydrochloric acid addition salts.

The acid addition salts below, when formed into an aqueous injectable preparation, provide the zwitterion in solution. The zwitterion has the structure according to formula 1 of the formula sheet.

The broad spectrum of activity against various organisms of the zwitterion form and thus of aqueous preparations made from the salts is described by the data in US patent 4,406,899. demonstrated. *

Aqueous preparations composed of the acid addition salts simply by the addition of sterile water yield acidic solutions which produce an unacceptable irritation when administered intravenously to rabbits and an unacceptably painful sensation when administered intramuscularly to rabbits.

The sulfuric and di-nitric acid addition salts have a reduced solubility that is insufficient for typical injectable preparations. It has been found herein to overcome these objectionable properties by using the salts herein in a physical mixture (which is a mixture of solids) with a pharmaceutically acceptable 3301991 * - 3 - non-toxic organic or inorganic base in amounts that when diluted with water to a zwitterionic activity of 1 mg / ml to 400 mg / ml, normally 250 mg / ml a pH of about 3.5 to about 7 is supplied (determined by high capacity liquid chromatography, 5 HPLC hereinafter).

A preferred salt is the crystalline sulfuric acid addition salt. This is preferred because its low water solubility (25 mg / ml) allows high recovery from the aqueous medium upon crystallization. The crystalline sulfuric acid addition salt is readily prepared by a process consisting of the steps of (a) forming a mixture of (i) at least one molar equivalent of sulfuric acid and (ii) zwitterion in an amount to make it present in the mixture at a concentration greater than 25 mg / ml, 15 (b) crystallizing the sulfuric acid addition salt and (c) isolating the crystalline sulfuric acid addition salt.

Fig. 1 is a graphic representation of the infrared absorption spectrum of crystalline 7- [a- (2-aminothiazol-4-yl) -a- (Z) -methoxyiDjina-acetamido] -3 - [(1-methyl-1-pyrrolidinio ) methyl] -3-cephem-4-carboxylate sulfate salt, measured at a KBr dilution thereof.

Fig. 2 is a graphical representation of the infrared absorption spectrum of the crystalline sesquiphosphate salt of 7- [a- (2-aminothiazol-4-yl) -x- (Z) -methoxyiminoacetamido] -3 - [(1-methyl-1-pyrrolidinio) methyl-3-cephem-4-carboxylate measured at a KBr dilution thereof.

FIG. 3 is a graphic representation of the infrared absorption spectrum of the crystalline diphosphate salt of 7- [a- (2-aminothiazol-4-yl) -x- (Z) -methoxy-iminoacetamido] -3 - [(1-methyl-1 pyrrolidinio-methyl] 3-cephem-4-carboxylate measured at a KBr dilution thereof.

The crystalline salts (hereinafter simply referred to as the salts) have excellent stability at room temperature and have an activity loss (as determined by HPCL) of less than 1% during one month of storage at room temperature. These salts also have excellent stability at elevated temperatures and an activity loss (as determined by HPLC) of less than 15% upon storage for one month at 45-46 ° C. The sulfuric acid addition salt is a preferred salt.

SSQ19S 1 - 4 - f>

It has an activity loss of less than 10% for a month when stored at 45-46 ° C. It is very important that it has a low water solubility / i.e. about 50 mg / ml so that it * is crystallized from water with minimal residue loss.

The di-nitric acid addition salt also has a low water solubility, i.e. about 60 mg / ml, and therefore also provides a low residue loss upon crystallization from water. The mono-hydrochloric, di-hydrochloric or sesqui or di-orthophosphoric acid addition salts have a water solubility greater than 200 mg / ml and are therefore preferably crystallized from organic solvents rather than water to obtain good yields .

The preparation of the salts is now discussed.

As previously indicated, the sulfuric acid addition salt is prepared by a process consisting of the steps of (a) forming a mixture of (i) at least one molar equivalent of sulfuric acid, (ii) zwitterion corresponding to said salt in such amount that it is present in the mixture at a concentration of more than 25 mg / ml, (b) allowing crystallization to take place and (c) isolating the crystalline sulfuric acid addition salt.

Preferably, the zwitterion in step (a) is used in an amount such that it is present in the mixture at a concentration of about 100 mg / ml to about 200 mg / ml, while step (b) is in an aqueous medium free of organic solvent is run. Normally no more than 2 molar equivalents of sulfuric acid are used in step (a). Zwitterion in step Ca) is normally used in an amount such that it is present in the mixture at a concentration of less than 500 mg / ml.

Step (a) is easily performed by adding either solid zwitterion to the sulfuric acid solution (eg IN 2 SO 2) with rapid stirring to form a solution or optionally performing step (a) by adding solid zwitterion in water and slowly add sulfuric acid with stirring to form a solution.

Step (b) is carried out by causing crystallization, preferably 199 1 · * - »Λ __" 3 by seeding and then preferably suspending for 15 minutes to 2 hours. It is preferred that this crystallization step be in an aqueous medium, free of organic solvent, is run and in such a case, purities greater than 98% are normally obtained.

While the presence of organic solvent, such as acetone, promotes crystallization and increases yield by lowering the solubility of the sulfuric acid addition salt formed in a crystallization medium, it can also promote impurity precipitation leading to reduced purity. When the zwitterion in step (a) is used in an amount such that it is present in the mixture in an amount of less than 25 mg / ml, an organic solvent, preferably acetone, must be included in the crystallization medium to ensure reasonable recovery to deliver. When acetone is used, it is suitably used in amounts of 0.5-10 parts by volume per volume by volume of aqueous crystallization medium.

Step (c) is performed by separating the crystals from the crystallization medium, preferably by vacuum filtration, then washing, e.g. with acetone / water, followed by acetone alone or Q, 1N 20 sulfuric acid (e.g. 1/10 vol.dl) followed by acetone (e.g. 1/1/4 volume part) and then drying, e.g. by vacuum drying at 30-50 ° C for 4-20 hours. The method of forming the sulfuric acid addition salt mentioned herein results in a purification of the sulfur ion form due to the limited solubility of the sulfuric acid addition salt compared to the zwitterion form and this can be used to purify the sulfur ion without isolating it as a solid . If it is desired to obtain substantially pure zwitterion (free base) from the sulfuric acid addition salt formed, this can be done by dissolving the salt in water, adding Ba (QH) 2.8 ^ 0 in an amount of 90-100% of the theoretical value at a pH of less than 6.5 to precipitate BaSO4, filter to remove BaSO4 and recover the filtrate, which contains a dissolved zwitterion and use it as a solution or solid zwitterion (free base) by freeze-drying or by adding acetone to precipitate amorphous zwitterion, followed by isolation of the solid zwitterion by vacuum filtration. 6 - tration, washing e.g. with acetone and vacuum drying. Optionally, the sulfuric acid addition salt is converted to the free base using ion exchange resins, e.g. Dowex WGR (weak basic anion exchanger resin) and Dowex XU-40090.01 (a strong acidic cation exchanger resin) with 5 subsequent freeze drying.

As for the preparation of the crystalline di-nitric acid addition salt; this is obtained by mixing (i) at least two molar equivalents of nitric acid and (ii) zwitterion corresponding to said salt in amounts such that it is present in the mixture at a concentration of greater than 100 mg / ml.

to mix and then cause crystallization by seeding or rubbing with a glass rod, diluting with 2-propanol and cooling. The crystalline di-nitric acid addition salt is collected, e.g. by filtration, washing sequentially, e.g. with 2-propanol (50% v / v), 2-propanol and ether and then drying under vacuum at 50 ° C for 2 hours.

The mono hydrochloric acid addition salt is prepared by dissolving the zwitterion in approximately one molar equivalent of hydrochloric acid and allowing crystallization to take place by adding acetone with stirring and continuing to stir, followed by isolating crystals, e.g. by vacuum filtration, then washing with acetone and vacuum drying. Optionally, the mono-hydrochloric acid addition salt is formed from the di-hydrochloric acid addition salt by suspending a di-hydrochloric acid addition salt in methylene chloride and adding one molar equivalent of triethylamine, followed by suspension to form the mono hydrochloric acid addition 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 is prepared by dissolving the zwitterion in at least two molar equivalents of hydrochloric acid, then allowing crystallization to take place by addition of acetone, then isolating the crystals, e.g. by vacuum filtration, washing with acetone and drying under vacuum. The crystalline di-orthophosphoric acid addition salt is prepared by dissolving the zwitterion in at least two molar equivalents of phosphoric acid, allowing crystallization to take place. by isolating 8601991-7 addition of acetone and the crystals by e.g. vacuum filtration followed by first washing with acetone and then with ether and then vacuum drying. The crystalline sesqui orthophosphoric acid addition salt is formed by the same procedure with the exception that about 1.5 molar equivalents of phosphoric acid are used. The salts are converted into injectable preparations by diluting with sterile water and buffering at a pH of 3.5-7 to form an injectable concentrate of 1 mg / ml to 400 mg / ml zwitterion. Suitable buffering agents include e.g. trisodium orthophosphate, sodium bicarbonate, sodium citrate, N-methylglucamine, lysine and L-arginine. For intramuscular or intravenous administration to an adult human, a total dose of about 750 to about 3000 mg / day in divided doses is normally sufficient.

It is not desired that the salts be converted to injectable preparations by simply adding sterile water because the sulfuric and di-nitric acid addition salts are not sufficiently soluble to form preparations of normal concentration for administration and because the salts mentioned herein, when dissolved , deliver very low pH preparations (1.8-2.5) that give a sore feeling when injected. As indicated above, it has been found herein that these drawbacks can be overcome by forming the salts herein into a physically, ie, a solid, mixture with pharmaceutically acceptable, normally solid, non-toxic organic or inorganic bases, in such amounts that a pH is from about 3.5 to about 7, preferably from about 4 to about 6, upon diluting the mixture with water to an injectable concentrate of 1 mg / ml to 400 mg / ml zwitterion, eg a zwitterionic activity of 250 mg / ml as determined by HPCL analysis. The precise proportions of the ingredients in the physical mixture vary from batch to batch of the salt, since the purity of the salt varies from batch to batch. The amounts of ingredients are determined for a given range by prefiltration with respect to a sample to obtain a selected pH within said range. The physical mixture is easily stored and transported in solid form, taking advantage of the stability of the salts and it is easily converted into an injectable 060 1 931 ν "ν - 8 preparation simply by adding water, eg by a sister or the doctor, precisely the use.

The physical mixture is prepared by mixing the salt and base into a uniform mixture, e.g. with a standard mixer in a dry atmosphere, after which it is preferably filled in a bottle or other container, all under aseptic conditions.

The bases for use in the mixture include, e.g. trisodium orthophosphate, sodium bicarbonate, sodium citrate, N-methylglucamine ,. I-1-lysine and -arginine-lysine and L-1-arginine are preferred, since mixtures thereof can be made into injectable preparations which produce less pain in animals when injected than preparations derived from mixtures that contain other bases.

(+)

The L-lysine is most preferably used in an amount providing a pH of 3.5-6 upon dilution of the mixture with water, to give a composition having a zwitterionic activity of 250 mg / ml (as determined by the HPLC analysis).

The salts, and the substantially dry physical mixtures containing them, can be stored without refrigeration or insulated packaging and still retain their high activity. In several of the compositions mentioned herein, the unstable zwitterion is used as the starting material. Its preparation is described in Examples 1-3 of U.S. Patent No. 4,406,899. The zwitterion is disclosed in said U.S. Patent as 7 - [(Z) -2-methoxyimino-2- (2-aminothiazol-4-yl) acetamido-3 - [(1-methyl-1-pyrrolidinium) methyl] -3- 25 cefem-4-carboxylate.

The invention is further illustrated by the following working examples.

EXAMPLE I

Preparation of the sulfuric acid addition salt.

1.5 g of Zwitterion is slowly added to 10 ml of rapidly stirred IN 2 SO 4 (1.59 mol equivalents) at 20-26 ° C. A dilution is obtained. Crystallization is then induced by seeding with crystalline sulfuric acid addition salt. after which the crystalline mass is suspended for half an hour. The crystals are then isolated by vacuum filtration, washed with 3 ml of a 50% acetone / water 1019 31-9 (v / v) and with 25 ml parts of acetone and at 40-50 ° C for dried under vacuum overnight. A typical yield is 1.3 g of sulfuric acid addition salt.

Analysis: Calculated for C19 H24 N6 ° 5S2 "H2 SO45% C" 39.44; % H '4 ^ 53; % N, 14.52; % S, 16.62; % H20, none.

5 Found:% C, 38 ", 91; % H, 4.57; % N, 14.64; % S, 16.71; % H 2 O, 1.42.

EXAMPLE II

Preparation of the sulfuric acid addition salt.

1.5 g of Zwitterion is dissolved in 5 ml of water. 5 ml of 1 M 2 SO 4 10 is slowly added to this solution while rotating. Crystallization is then started by seeding with crystalline acid addition salt, after which the crystalline mass is suspended for half an hour. The crystals are then isolated by vacuum filtration, washed with 3 ml of 50% acetone / water (v / v) and with a 25 ml portion of acetone and dried under vacuum at 40-50 ° C overnight.

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

EXAMPLE III

Preparation of the (HNO2) 2 acid addition salt.

300 mg of zwitterion were dissolved in 2N nitric acid (0.5 ml).

The solution is triturated with a glass rod, diluted with 2-propanol (0.4 ml) and cooled. The crystalline title compound is collected and washed successively with 0.4 ml of 2-propanol E 2 O (1: 1), 2-propanol and then ether to provide 127 mgs of the dinitrate salt.

Analysis: Calculated for C 8 H N-O S 2 HNO:% C, 37.62; % H, 4.32; 19 24 6 5 2 3% N, 18.47; % S, 10.57;

Found:% C, 36.92; % H, 4.10; % N, 18.08; % S, 10.67; (^ 0 content 0.90%).

EXAMPLE IV

Preparation of the Monohydrochloride Acid Addition Salt.

1 g of Zwitterion is dissolved in 2.08 ml in 1N HG1 (1 molar equivalent) at 20-25 ° C. 30 ml of Acetone are added with rapid stirring over a 15 minute period to form crystals.

850 1 931

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- 10 -

Stirring is continued for 1 hour. The crystals are isolated by vacuum filtration, washed with 10 ml of acetone and dried under vacuum at 50 ° C for 2 hours. A typical yield is 0.9 g of crystalline monohydrochloride salt.

Analysis: Calculated for C.X.N.O.S. HC1:% C, 41.37; % H, 4.75; 19 24 6 o 2% N, 15.2; % S, 11.63; % Cl, 12.86

Found:% C, 39.32; % H, 4.88; % N, 13.95; % S, 11.28; % C1, 12.44; % H 2 O, 4.5.

(Adjusted for ^ 0:% C, 41.17;% N, 14.61;% S, 11.82; 10% C1, 13.03).

EXAMPLE V

Preparation of the dihydrochloride acid addition salt and preparation of the monohydrochloric acid addition salt therefrom.

350 mg of Zwitterion is dissolved in 2 ml of 1N-HCl. 10 ml of Acetone 15 are added to the resulting solution with rapid stirring over a period of 5 minutes to form the crystals.

Stirring is continued for an additional 5 minutes. Then, additional 10 ml of acetone are added and stirring is carried out for 0.5 hour.

The crystals are removed by vacuum filtration, washed with 25 ml portions of acetone and dried under vacuum at 40-45 ° C for 24 hours. A typical yield is 300 mg of crystalline dihydrochloride acid addition salt.

Analysis: Calculated for: cigH24N6 ° 5S2 · 2HCl:% C, 41.38; % H, 4.75; % N, 15.2; % S, 11.62; % C1, 12.8.

Found:% C, 40.78; % H, 4.98; % N, 14.7; % S, 11.25; % H 2 O, 1.25.

(Adjusted for H 2 O:% C, 41.1;% N, 14.88;% S, 11.39;% C1, 11.94).

1 g of the dihydrochloride salt as prepared above is suspended in 20 ml methylene chloride at 20-25 ° C in a sealed flask and 0.28 ml of triethylamide is added over a period of 15 minutes. The crystalline mass is then suspended with stirring for 5 hours. The monohydrochloride crystals obtained are then isolated by vacuum filtration, washed with 25 ml parts of methylene chloride and 360 1 99 L-3-lien dried under vacuum at 50 ° C for 2 hours. Typical yield is 800 mg.

EXAMPLE VI

Preparation of the di-orthophosphoric acid addition salt.

1 g of Zwitterion is dissolved in 3.4 ml of 144 mg / ml H 2 PO 4 (2.2 molar equivalents) at 15 ° C. The resulting solution is filtered appropriately to make it clear. 12 ml of Acetone are added to the purified solution with rapid stirring over a 10 minute period to form crystals. Stirring is continued for 10 minutes. Then 30 ml of acetone are added over a period of 10 minutes and stirring is continued for an additional 15 minutes. The crystals are collected by vacuum titration, washed with 25 ml parts of acetone and 25 ml parts of ether and dried under high vacuum for 16 hours. A typical yield for this type of preparation was 1.1 g of crystalline di-orthophosphoric acid addition salt.

Analysis: Calculated for .2Η ^ ΗO4:% C, 33.72; % H, 4.47; % N, 12.42.

Found:% C, 33.43; % H, 4.65; % N, 12.02; % H 2 O, 1.82. (corrected for ^ 0:% C, 34.0;% N 12.2).

The sesqui orthophosphoric acid addition salt is formed as above except that 1.5 molar equivalents of H 2 PO 4 are used instead of 2.2 molar equivalents.

EXAMPLE VII

Stability at elevated temperature.

Increased teraperature stabilities were determined by storing the preparations in dry containers at temperatures and for periods of time as specified below, with the activity losses or increases determined by HPLC. A percent activity gain is indicated by a plus sign before a number. Less than 10% activity loss over a 2-4 week period at 45-46 ° C is usually indicative of less than 10% activity loss over a 2-3 year period at room temperature.

8601991 - 12 - -ίV '

PERCENTAGE LOSS

45 ° C - 56 ° C 100 ° C

(weeks) "(weeks) (days)

Form 1 '2 4 6 1' 2 '4 1 5 Zwitterion 37 5171 - 57 - - 100 H-SO-SOUT 2.4 - + 5i 3 +5; 1.4 5 - +6! +3 0 - +6: 0 - 10

2 4 ί i I

I! i .......- ---------- ----------------. | 1 | | - · T · - “- t - L - - ........- - -1 -----; -------- (HN03) 2 salt 8.8 3.4 0, 68 10.3 y 3.7 y 2.4: - - ^^ - y - ^ - HC1 salt 4.8 2.3! 6.0 6.4 6.4 y - i i

"τ · T r" · rr '1 ~~ - "t T I

I! (HCl) salt 0-7.4; - 0; - 7.2 i 12.4 __ ^ ___._ i_I_! 10 (H3PO4) 2 salt 0 3.0 1.0 - 2.7 5.0 -;

EXAMPLE VIII

Investigation of physical mixtures.

Physical mixtures were composed of crystalline sulfuric acid addition salt with (a) trisodium orthophosphate, (b) sodium bicarbonate, (+) (+) 15 (c) L-lysine and (d) L-arginine.

The bases were added in amounts to obtain a pH upon dilution of the mixture with water to a zwitterionic activity of 250 mg / ml (as determined by HPLC analysis) as follows: trisodium orthophosphate (to obtain a pH of 6.0 to give)? Sodium bicarbonate (to give a pH of 6.0); L-lysine (to provide a pH of 6.0); L-arginine (to provide a pH of 6.0).

Injectable preparations were formulated by making to a zwitterionic activity of 250 mg / ml with sterile water as determined by HPCL analysis. There were no solubility problems.

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The injections (100 mg / kg) were made intramuscularly to rabbits with pain within an acceptable threshold. The least pain was caused by the arginine-containing preparation. Similar results of good solubility and acceptable pain on intramuscular injection were obtained using the other salts mentioned, physically mixed with the above bases.

Fig. 1 is the infrared absorption spectrum of the crystalline sulfate salt prepared as described in Examples I or IX pelletized in the crystalline form with potassium bromide. The X-ray diffraction pattern 10 of the crystalline sulfate salt of 7- [a- (2-aminothiazol-4-yl) -a- (Z) -methoxyiminoacetamido] -3 - [(1-methyl-1-pyrrolidinio) -methyl] -3- cefem-4-carboxylate, prepared as described "in Examples I or II, was determined with a Rigaku Powder diffractometer, using a copper X-ray target and a nickel filter while the sample was contained in a glass disc. 2 ° / minute over a range from 5 ° to 40 ° and a graph was mechanically recorded to show the angles of maximum diffraction, from which the (d) distances and relative (1/1 °) were calculated, they are summarized below .

d distance (° &) 1/1 ° (%) 20 9.20 100 6.80 50 5.50 28 5.09 22 4 ^ 50 38 25 4.41 44 4.19 63- 3.78 38 3, 64 44 3.39 25 30 3.31 31 3.15 47 8 SÖ1991

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EXAMPLE IX

Preparation of the sesqui phosphate salt.

The zwitterion, 0.70 g, is dissolved with rapid stirring in 2.2-2.4 ml of 85% phosphoric acid (2.1-2.2 molar equivalents diluted 1:10 (v / v) 5 with water The solution is clarified by filtration through a 0.22-0.45 micron pore size membrane filter 5-7 parts by volume (15-20 ml) of methanol are stirred under rapid stirring for a period of 30-60 minutes. filtrate added During this treatment crystals form, continuing the rapid stirring for 1.5-2 hours 10. The crystalline product is recovered by vacuum filtration. The product is washed on the filter with 6-8 ml of 1: 1 (v / v) methanol: acetone, taking care to maintain a densely packed filter cake, then with acetone The product is dried under vacuum at 50 ° C for 2 hours, a typical yield is 0.7-0 75 g.

Infrared interpretation (see figure 2) (IR, KBr pellet)

Peak position (cm "1) Functional group

2800-3400 NH, NH3 +, carboxyl OH

1780 β-lactam C = 0 1680 carboxyl C = 0 1660 amide C = 0

1630 C-N, C = C

1550 amide OH

25 980.1040 PO = 4

Behavior when heated.

There appears to be an exotherm at 171.8 ° C according to differential scanning calorimeter graph.

X-ray diffraction pattern.

The X-ray powder diffraction pattern of the aforementioned sesquiphosphate salt was measured with a Rigaku powder diffractometer in the same manner as described above for the sulfate salt, the results of which are as follows: 860190f - 15 ι / ,0 ° 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 NMR interpretation (H 90 MHz NMR, D 2 O solution) see formula 2 of the formula sheet.

Chemical shift Description Total Assignment 2.0-2.4 Multiplet 4 14CH2, 14'CH2 20 3.04 Singlet 3 12CH3 3.3-3.6 Multiplet 5 2CH, 13CH2, 13'CH2

3.94 Doublet 1 2CH

4.12 Singlet 3 20CH3

4.12 Doublet 1 11CH

25 4.8 Doublet 1 11CH

5.42 Doublet 1 6CH

5.88 Doublet 1 7CH

7.21 Singlet 1 18CH

) 01991 - * ü '- 16 -

Stability

Time-temperature percentage loss 1 day; 100 ° C - 10.9 - 3 days; 70 ° C - 0 5 7 days; 70 ° C - 1.9 1 week; 56 ° C - 1.0 2 weeks; 56 ° C - 1.4 weeks; 56 ° C - 0 1 week; 45 ° C 0 10 2 weeks; 45 ° C -1.4 weeks; 45 ° C - 0.7 8 weeks; 45 ° C - 1.6 1 month; 37 ° C - 2.5

Elemental analysis (weight percent) 15 Found Dry basis theory (Sesqui phosphate) C 35.44 36.3 36.4 H 4.66 4.41 4.7 N 12.88 13.2 13.4

H ^ O 2.29 * - monohydrate = 2.8% H ^ O

20 H3PO4 23.06 23.6 23.6 * Karl Fisher Method 1601 991

Claims (2)

1. Temperature stable crystalline salts of 7- [α- (2-amino-thiazol-4-yl) -a- (Z) -methoxyiminoacetamido] -3 - [(1-methyl-1-pyrrolidinio) -methyl] -3-cephem-4-carboxylate selected from the group consisting of the sulfuric, di-nitric, mono-hydrochloric and di-hydrochloric acid addition salts and orthophosphoric acid addition salts, which have 1.5-2 molar equivalents of H -.PO. contain or solvates thereof. 3 4 Crystalline salts according to claim 1, selected from the group consisting of sulfuric, mono-hydrochloric, di-hydrochloric and orthic hydrochloric acid addition salts or sulfates thereof, The physical mixture of the salt of claim 1, having a pharmaceutically acceptable, non-toxic organic or inorganic base in proportions, upon dilution of the mixture with water to injectable concentration a pH of about 3.5 to about 7 will be delivered. Physical mixture according to claim 3, characterized in that the salt and base are present in amounts, upon dilution of the mixture with water to an injectable concentration providing a pH of about 4 to about 6. Physical mixture according to claim 4, characterized in that the base is L (+) lysine. Physical mixture according to claim 4, characterized in that the base is L (+) arginine. 7. Process for making the sulfuric acid addition salt, characterized by the following steps: (a) forming an aqueous mixture of (i) at least one. molar equivalent of sulfuric acid and (ii) zwitterion corresponding to said salt, (b) crystallizing the sulfuric acid addition salt, provided that when a zwitterion is present in the mixture at a concentration of less than 25 mg / ml, the crystallization in the presence of an organic solvent is performed and (c) isolating the crystalline sulfuric acid addition salt. 860 1 991 ./ - 18 - Process according to claim 7, characterized in that step (b) is carried out in an aqueous medium free from organic solvent. Process according to claim 8, characterized in that the Zwitterion in step (a) is used in an amount such that it is present in the mixture at a concentration of less than 500 mg / ml. A method according to claim 7, characterized in that the amount of the zwitterion used in step (a) is such that it is present in the mixture at a concentration greater than 25 mg / ml. A method according to claim 8, characterized in that the zwitterion in step (a) is used in an amount such that it is present in the mixture at a concentration of about 100 mg / ml to about 200 mg / ml. Crystalline sulfuric acid addition salts according to claim 1. 13. Crystalline 7- [a- (2-aminothiazole) -4-yl) -a- (Z) -methoxy-iminoacetamido] -3-¾ (1-methyl-1- pyrrolidinio) methyl] -3-cefem-4-carbo-xylate sulfate salt with the following X-ray powder diffraction pattern: d distance (° &) 1/1 ° (%) 9.20 100 20 6.86 50 5.50 28 5.09 22 4.50 38 4.41 44 25 4.19 63 3.78 38 3.64 44 3.39 25 3.31 31 30 3.15 47 A crystalline orthophosphoric acid addition salt according to claim 1 and the hydrates thereof. ^: 1991 - 19 - 15. Crystalline 7- [α- (2-aminothiazol-4-yl) -ct- (Z) -methoxyimino-acetamido] -3 - [(1-methylpyrrolidinio) -methyl] -3-cephem-4-carboxylate phosphate with the following X-ray powder diffraction pattern: d1 / 1 ° 5 11.04 32 9.2 16 7.89 - 24 7.02 - 42 6.7 - 32 10 5.5 - 26 4.64 - 100 4.456 53 4.3- 58 3.88 - 26 15 3.75 - 89 3.56 - 21 3.31 26 3.05 - 16 860 1 991 N — ΓΓ — C-CONH — I- H k _ f / k 2 0_ch3 X © 2 M ch3 20.n / 0CH3 / Kj / ix / 1111 v 13 NH / 5 \ ® / Υ * 2. J o jy — k Yk λ 3 ^ 8 ^ γγ \ Jw io CO®, 3 ^ 360 1 90 1