IE862078L - Cephalosporin salts - Google Patents

Abstract

Cephalosporin Salts and Injectable Compositions Crystalline sulfuric, di-nitric, mono-hydrochloric, dihydrochloric, and di- and sesqui-orthosphosphoric acid addition salts of 7-¢.alpha.-(2-aminothiazol-4-yl)-.alpha.-(Z)-methoxyimino-acetamido!-3-¢(1-methyl-1-pyrrolidinio)-methyl!-3-cephem-4-carboxylate are stable even at elevated temperatures. The crystalline sulfuric acid addition salt is made by forming an admixture of (a) at least one molar equivalent of sulfuric acid with (b) zwitterion in an amount so as to be present in the admixture at a concentration of greater than 25 milligrams/ml, causing crystallization, separating the crystals, washing and drying. The crystalline monohydrochloride, dihydrochloride, and orthophosphate salts are prepared by dissolving the zwitterion in the appropriate amount of acid, causing crystallization by adding acetone and isolating the crystal. Physical admixtures of the salts with certain bases in proportions to give a pH ranging from about 3 5 to about 7 on dilution with water provide injectable compositions on dilution. Brief Description of the Drawing Figure 1 is a graphical representation of the infra red absorption spectrum of crystalline 7-¢.alpha.-(2-aminothiazol-4-yl)-.alpha.- (Z)-methoxyiminoacetamido!-3-¢(1-methyl-1-pyrrolidinio)methyl!-3-cephem-4-carboxylate sulfate salt measured on a KBr dilution thereof. BMY-28142.H2SO4 - IR Spectrum. [CA1284994C]

Description

59222 ss The invention relates to cephalosporin salts and injectable compositions.

Crystalline sulfuric,, di-nitric, Bono-hydrochloric, di-hydrochloric, and di- and sesqui-orthophcsphcria acid addition salts of 7-l«-g2-aminothiazol-4-ylS-a- Figure 3 is a graphical representation of the infra red absorption spectrum of the crystalline diphosphate salt of 7- [a™ (2-auiinothiazol>4-yl) -x- (Z) -raethoxyiminoacetamido ] -3-[(1-methyl-l-pyrrolidinio)-methyl3-3-cephem-4-carboxylate 10 measured on a KBr dilution thereof. « 6 4 Thie invention is directed to temperature stable semi-synthetic cephalosporin salts whose preparation has not been described in the literature, to the preparation of such salts, and to admixtures containing these salts,, 5 Aburaki et al. 0„S„ Patent &3o. 4,406,899 discloses 7- |©- C2-aminothiaaol~4-yl|-c-C2i-methQxy iminoacetamido] -3-I {1-snethyl-l-pyrrolidinio)-methyl]!-3-cephem—4-carboscylate in the zwitterion for,® and mentions corresponding acid addition salts {which as© present in the switterion form in injectable composi-10 tionsj) ani shows that the switterion form has broader spectrum activity than ceftazidime and cefotaxime.

However, th® aforementioned Ahuraki jet al. cephalosporins are ©table only for a few hours as injectable compositions and the zwitterion form even as a dry powder is unstable at room 15 temperature and loses 30% or more of its activity on storage at elevated temperatures (e.g. 45 deg. C and aboveS for even on© week and therefore requires special insulated packaging and/or refrigeration and is at a packaging and storage disadvantage compared to ceftazidime and cefotaxime. 20 Hhile Aburaki et al. mentions acii3 addition salts, the patent doss not state how to sake these or state which if any of these salts have good stability in dry powder form. Kessler et al., "Comparison of a New Cephalosporin, BK¥ 28142, with Other Broad-Spe ctrum -Lactam Antibiotics", Antimicrobial Agents and 25 Chemotherapy, Vol. 27, No. 2, pp„ 207-216, February 1985 mentions the sulfate salt, but ctoes sot disclose bow to prepare such or that this salt has room temperature stability and good elevated taiffipesratv,re stability in flry powder form* It has bees discovered herein that certain crystalline acid addition salts of 7-Ja- ^-aminothiasol—fl-ylj^e-|Si -inethoxyliminoBcetan1doV~3~ I (l-methyl-l-pyrroli<3ini©) -methyl])-B-eephem-^-earboscylate in dry powder for a have excellent room 5 temperature stability and have superior elevated temper&ture iKtatoility compared to the zwitterion 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 10 7- (2-asninothiazol-4-yl}-a- (2)-methoxyiminoacetamido]~3-I (l-methyl-l-jayrrolidinio} -snethylJ-3-cepfaem-4-carboxylate selected from the sulfuric, di-nitric, mono-hydrochloric, and di-hydrochloric aciel addition salts and orthophosphoric acid addition salts (1.5-2 soles of 15 ortli©phosphoric acid per snole of salt,, e.g. a range of from the sesqui- to the di-orthophosphoric acid salts}, or solvates thereof.. The term "crystalline" is used, herein to mean at least some characterising arrangement of molecules. While the sulfuric, di-nitric, di-hydrochloric and orthophosphoric acid 20 addition salts herein are prepares in clearly ervstslline form (as evidenced by birefringence under a polarising microscope) with precise arrangement of molecules,* the mono-hydrochloric aeii addition salt, has been prepared only with some regularity in the arrangement ©f its molecules (as evidenced by poor birefringence 25 ender polarising microscope} and not a precise preiiot&ble arrangement and thus is '"poorly" crystalline. The term "crystalline" is used herein to embrace sot only the clearly crystalline salts but also the "poorly" appearing crystalline pono-hydrochloric acid (addition salt- " 30 The acid addition salts herein when formed into aqueous injectable compositions provide the zwitterion in solution. The zwitterion has the striae** The broad ispectr&an utility Against various organisms of the zwitterion form® and thus of aqueous compositions made up from the salts herein, is shown by the data in M>ur&ki et al. U.S., 4,406,899.

Aqueous composition© made ■asp from the acid addition ©alts feereiai siiaply by the addition ©f sterile water provide acidic solutions which provoke unacceptable irritation ©n intravenous administration to rabbits and en&eceptahly painful sensation on intramuscular administration to rabbits. The sulfuric acid and dli~nitrie acid addition salt© have reduced solubilities which are insufficient for typical injectable compositions. St has been found herein that these objectionable characteristics are overcome by utilising the salts herein in physical admixture |that is as an admixture of solids! with a pharmaceutics lly acceptable non-toxic organic or inorganic has® in proportions to provide a pB of about 3.5 to 7 on dilution with water to a switterion activity of from 1 tag/ml to 400 tag/ml, norms lly 250 ag/aal las determined by high performance liquid chromatography, he re ins ft© r HPLC). h preferred salt herein is the crystalline sulfuric acid addition salt. It is preferred because its low solubility in water |25 sag/ml) allows high recovery from aqueous sftediuin on crystallisation. the crystalline sulfuric acid addition salt is readily prepared by a process comprising the steps of fail forming an aqueous admixture of Cil at least 1 molar equivalent of sulfuric acid and till zwitterion in an asaount so as to be present in the admixture at a concentration greater 'than 25 tng/ml, t'b! causing crystallisation of the sulfuric acid addition salt to occur,, and m isolating crystalline sulfuric acid addition salt.

The crystalline salts herein (hereinafter referred to ©imply ■*® the salts herein) have excellent stability at room temperature and hav© a potency loss |as determined by HPLCi, of less than 1% csa storage for a month at room temperature. These salts also have excellent stability ©t elevated temperatures and have © potency loss las determined toy HPLCi of less than 15% on storage for a month at 45-56 deg. C.

The sulfuric acid addition salt is a preferred salt herein.

It has a potency loss of less than 10% on storage for a month a 45-S6 deg. C. Very importantly, it has a low solubility in veter, i.e. About 2S mg/ml, ani therefore is crystallised from vater with minimised residual loss.

The di-aitric acid addition salt herein also has a low solubility in water, i.e. about 60 ng/snl, and therefore also provides low residual loss on crystallization from water.

The raono~ hydrochloric, di-hydrochloric and sesaui- or (di-ort-hophosphorie acid addition salts have crater solubilities greater than 200 mg /sal, ,, am el therefore are preferably crystallised from organic solvents, rather than from water, in order to obtain good yields.

We turn no%? to the preparation of the salts herein.

As previously indicated the sulfuric acid addition salt feerein is prepared by a process comprising th® steps of fa) forming aa aqueous admixture of fil at least one molar equivalent of sulfuric acid and Cii! zwitterion corresponding to said salt in an amount as to be present in the admixture at a concentration of greater than 25 mg/ml, fhl causing crystallization to occur,, anfl (cj iisoiatimsg crystalline sulfuric acifl afiflition salt. 8 Preferably the zwitterion is used in step (a) in an amount bo as to be present is the Admixture at a concentration ranging from about 100 sg/aal to 200 wig/ml* and step (b) is carried out is -an aqueous s®edi°am free of organic solvent. IMoraaliy no snore 5 than 2 solar equivalents of sulfuric acid are utilised in Etep fa!. Norma lly zwitterion is used in step (a} in an amount so as to be present in the admixture at a concentration less than 500 sag/ml.

Step fa J is reaflily carried out either by adding solid 10 2witterion to sulfuric acid solution (e.g. IN B^SO^i with rapid stirring to form a solution. Alternatively step (a) can -be carried by dissolving solid zwitterion in water and slowly adding sulfuric acid with stirring to form a solution.

Step (h} xb carried out by inducing crystallization, 15 preferably by seeding,, and then slurrying, preferably for 15 minutes to 2 hours. St is preferred that this crystallisation step he carried out in aqueous medium,» free of organic solvent, and in such case purities greater than 98% are normally obtained. While the presence of organic solvent, such as acetone, fosters 20 crystallisation and increases yield by lowering the solubility of the formed sulfuric acia addition salt in the crystallisation medium, it also can foster precipitation of impurities resulting in decreased purity. Khen the zwitterion is used in step (aJ in an amount so as to he present in the admixture in an amount less 25 than 25 mg/ml, organic solvent, preferably acetone, must be included in the crystallisation medium to provide reasonable recovery., When acetone is used, it is appropriately used in amounts of 0.5 to 10 volumes per volume of .aqueous crystallization siedium„ 30 Step ifoi is carried out by separating the crystals from the crystallization medium, preferably by vacuum filtration, then washing e.g. with acetone/water followed by acetone alone or 0.1N sulfuric acid (e.g. 1/10 volume) followed by acetone (e.g. 1/4 volume), and then dryinge.g. by vacwua drying at 30-50 deg0 C for 4-20 b©ur&„, The saethod herein for forsisg the sulfuric acid ©flflition salt result© in th© purification off the zwitterion for® because S of the limited solubility of the sulfuric acii addition salt compares to 'the zwitterion form and can be used to purify zwitterion without isolating it as. a solid,, If it is desired to obtain substantially pure zwitterion (free-base) from the formed sulfuric aciffl addition salt, this can fee carried out by 10 dissolving the salt in water, adding BafOH^.SH^O in an amount of 90-100% of theory at a pB of less than S„5 to precipitate BaSO^, filtering to remove the BaSO^ and recovering the filtrate containing the zwitterion dissolved therein and utilising it as a solution or isolating solid zwitterion (free-base) by 25 lyophilizing it or by adding acetone to precipitate amorphous Zwitterion followed by isolating solid zwitterion by vacuum filtration, washing e.g. with acetone, and vacuum drying. Alternatively, the sulfuric acid addition salt is converted to the free-base utilising ion exchange resins? e.g. Dowex WGR {a 20 weak base anion exchange resin) and Dowex XU-40090.01 |a strong acid cation exchange resin! with subsequent lyophiliz at ion.

Turning now to the preparation of the crystalline di-nitric acid addition salt herein, this is obtained by admixing (i) at least two aolar equivalents.of nitric acid and fiil zwitterion 23 (Corresponding to said ©alt ®o to be present in the admixture at a concentration greater than 200 ng/sal, and then inducing crystallisation' by seeding or rubbing with a glass rod, diluting with 2-propanol and cooling. The crystalline di-nitric acid addition salt is recovered e.g- by filtering„ washing 30 sequentially, e.g* with 2-propanol-H^O 150% v/v) , 2-propanol,, and ether® and then vacuum drying at SO deg» C for 2 hoars. <8 The mono-hydrochloric acii addition salt herein is prepared by dissolving zwitterion is approximately one siolar equivalent of 1 0 hydrochloric acid and causing crystallisation by adding acetone with «tarring and continuing to stir* followed by isolating exystals, e.g. by vacuum filtration followed by washing with acetone a^d vacuum drying. Alternatively the aono-hydxochloric 5 acid addition salt is formed from the di-hydxochlorie acid addition salt by slurrying the di-hydrochloric acifi addition salt in methylene chloride and adding 1 stole equivalent of txiethylamine followed by slurrying to form the nono-hydrochloxic acid addition salt which is isolated® e.g. by ■vacuum filtration, 10 followed by washing with methylene chloride and vacuum drying.

The crystalline di-hydrochloric acid addition salt herein is prepared by dissolving zwitterion in at least two ®olar equivalents of hydrochloric acid® then causing crystallisation by adding acetone,, then isolating crystals e.g. by vacuum 15 filtration, washing with acetone and vacuum drying.

The crystalline di-orthophosphoxic acid addition salt herein is prepared by dissolving the zwitt&xion in at least 2 molar equivalents of phosphoric acid, causing crystallisation by adding acetone, and isolating crystals by e.g. by vacuum filtration 20 followed by washing first with acetone and then with ether and then vacuum drying. The crystalline sesgui-orthqphosphoxic acid addition ©alt is formed by this same procedure except that about 1.5 molar equivalents off phosphoric acid is esei.

The salts herein are formed into injectable compositions by 25 diluting with sterile water and buffering to a pS of 3.5-7 to form an injectable concentration of 1 sig/ml up to 400 asg/anl of zwitterion. Suitable buffering agents include, for example, trisodium orthophosphate, sodium bicarbonate, ®odimm citrate* Hmethylglucamine, L(+? lysine and L(+) arginine. For 20 intramuscular or intravenous administration to »>a adult human, a totai 1 dosage of from about 750 to about 3000 ag per The physical admixture is readily stored and! shipped in solid form thereby taking advantage of the stability of the salts herein and is readily converted into an injectable composition simply by addition of water* e.g. by a snurse or doctors, just prior to use.

The physical admixture is prepared by blending the salt &nfi the base into a uniform blend* e.g. utilizing a standard blender i» a dry atmosphere* and is then preferably filled into a vial or other container,, all under aseptic conditions.

The bases for use in the admixture include* for example * trisoflium orthophosphate* sodium bicarbonate*, sodium e£trste# N-me> thy Iglucamine» il+jf lysine and t(+) arginine. iW lysine and Lt4ji arginine are preferred sine© admixtures ©ont&ining these are reconstituted to provide injectable compositions which on injection provSde less pais in animals than compositions derived £xom admixtures containing other bases. The LI*) lysine is very preferably utili$ed in a proportion to provide * pH of 3.5-6 on dilation of the admixture with water to provide a composition with a zwitterion activity of 250 tag/ml (as determined by HPLC essay!.

The salts herein and substantially dry physical admixtures containing them can be stored without refrigeration ox insulated packaging and still retain high potency.

In several of the preparations herein the unstable zwitterion is used as the starting aaterial. The preparation of this is described in Examples 1-3 of Abwraki et al. U.S. 4,,406*889. Th© zwitterion is referred to in j&uraki et al. as 7-1 C2}-2-raethoxyimino-2- (2-aminothia;eol-4-yl Jacetaraido-3-1 (1-sne thy1-1-py rr ol idin ium) me thyl 3 -3~cephem- 4-carboxyl ate.

Th® invention is illustrated in the following working ©samples.

Exaanole I Preparation of the Sulfuric Acid Addition Salt 1.3 g of zwitterion are aafified slowly to 10 ml o£ rapidly stirred IN H950^ (1.59 molar equivalents$ at 20-26 deg. C. A solution is obtained. Crystallisation is then induced by seeding with crystalline sulfuric acid addition salt and the crystalline sass is slurried for 0„S hours. The crystals fere then separated by i?acwwKi filtration, washed with 3 al of 50% acetone/water (V/V) and with two 5 al portions of acetone, and vacuum dried at 40-50 deg« C overnight.

A typical yield 16 1-3 9 ©£ sulfuric acid addition salt.-Analysis: Calculated for c^S24Hg05S2J2SD4! *Cr 39-44' 1H» 4.53; IN, 34.52; %S( 16.62; tB^O, none. Founds 1C, 38.91; tH, 4.57;'%N, 14.64; %S, 1®."?!^ ULgO, 3.42.

Example II Preparation of the Sulfuric Acid Addition Salt 1.5 g of zwitterion are dissolved in 5 sal of *ater. 5 ml of 1M HgSOa are slowly added to this solution with stirrinq. Crystallization is then induced by seeding with crystalline acid addition salt and the crystalline mass is slurried for 0.5 hours* The crystals are then separated by vacuum filtration, washed with 3 »1 of 50% acetone/water (V/V) and with two 5 ml portions of acetone, <&ni vacuum dri®a at 40-50 deg. C overnight.

A typical yield is 1.3 g o£ sulfuric acid addition salt.

Example III Preparation of the Acid Addition Salt * ri~ a. -. ■ 300 sag ©£ zwitterion are dissolved in 2N nitric acii CO.5 ml J - The solution is rubbed with a glass rod, diluted with 2-propanol (0-4 «1) cooled. The crystalline title compound is collected and is sequentially washed with 0-4 ml of 2-propanol H»0 <1:1). 2-propamol ®,'n<3 then ether to afford 127 sags of the <& dinitrate salt.

Analysis: Calculated for C,2HKO^: %C, 37.62? «H, 4.32s %M, 18«, 47? %S, 10.57. Founds %C, 36.92; %H, -4.10; %N, 18.08^ 18, 10-67? (E^O content 0.90%). i 4 B* ample 3V Preparation of the Honohydroehloride Acid Addition Salt 1 g of zwitterion is dissolved in 2.08 ml of IN BC3L (l aoi&r equivalent) et 20-25 deg. C. Thirty ml of acetone are added with 5 rapid stirring over a 15 minute period whereby crystals forsm.

Stirring is continued for 1 hour. The crystals are isolated by vacuum filtration, washed with 10 sal of acetone and vacuum fiyiad at 50 dleg„ C for 1 hours.

A typical yield is 0.9 g ©f crystalline monohydrochloride 10 salt. Analysis! Calculated for CiaSLaNfi0~S,.9Cl: %C, 41.37? %H, 4,73s W, 15.2i %S, 31-63; %C1, 32.86. Founds %C, 39.32? «H, 4-88? %N, 33.95» %S„ 33.28? %C1, 32.44? IHjO, 4.5.

|Corrected for B,Os %C, 41.37,? %N, 34.63? %S, 31.82? %C1, 13.031.

Ex ample V 15 * Preparation of "the Dihydrochloride Acid Addition Salt end Preparation ©£ the Monohydrochloride Jicid addition Salt From It 350 sag of zwitterion are dissolved in 2 ml of 1N-HC1. 10 ml 20 of acetone are added to th© resultant solution, with rapid stirring and over a 5 minute interval, whereby crystals form. Stirrincj is continued for 5 additional tuinutes. Then 10 additional ml of acetone are added and stirring is carried out for 0.5 hours. The crystals are removed by vacuum filtration,, washed with two 5 al 25 portions of acetone and vacuum dried at 40-45 deg. C for 24 hours.

A typical yield is 3DQ tag of erystalline dihydrochloride acid addition salt. Ana3ysis Calculated for CSli^W2'23Cl! %C* 41'3Bs 4°15' m* 15.2? IS, 11.62? tea, 12.8^ ^Founds 1C, 40.78; IHf 4.98; IN, 14.7; IS, 11.25; IHjO. 1.25. |Corrected for B2©s IC, 41.1; IN, 14.88; «S, 12.39; IC1, 11.94). % g of dihydrcchloride salt prepared as above is slurried in 20 sal of methylene chloride at 20-25 deg. C in a sealed flask and 0*28 sal of triethylamine is added over a 15 minwte interval. Th® crystalline mass is then slurried for 5 hours- The resultant Esonohyetrochiorida crystals are then isolated by vacuum filtration* washed with two 5 sal portions of methylene chloride and vacuum dried at SO C for 2 hours. A typical yield is 800 aig.

Example ¥1 Preparation of the Di-orthophosphoric Acid Addition Salt 1 g of zwitterion is dissolved in 3.4 sal of 144 sag/ml B-.PO. <2.2 saolar equivalents) at 15 deg. C. The resulting 3 4 solution is suitably filtered to clarify it. 12 ml of acetone are added to the clarified solution, with rapid stirrino and over a 10 minute period ,tdiereby crystals form. Stirring is continued for 10 minutes. Then 30 ml of acetone are added over a 10 minute period* anol stirring is continued for an additional 15 minutes.

The crystals are collected by vacuum filtration* washed with two 5 al portions of acetone and two 5 al portions 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 C,2H3P04S %c' 33-*2; tB, 4.47; tN, 12.42. Found: ""iC, 33.43^ 11, 4.65; IK, S2..,02^ IH-jO, 1.82. fCorrected for 34.®? IN* 12.2K 1 8 The sesqui-Grthcphosphoric aeifi addition salt is formed as above except that 1.5 »olar equivalents of H3P04 are used instead of 2.2 siolar equivalents.

Example VI1 5 Stabilities at Elevated Temperatures Elevated temperatare stabilities vere determined by storing the preparations 1b dry containers at temperatures and for time periods as denoted below and potency losses or gains were determined by BPLC. A 1 potency gain is indicated by a plus sign 10 in front of a figure- A less than 10% potency loss over a 2 to 4 w&ek period at 4S-5S Steg* C is usually indicative of less than 10% potency loss over a 2-3 year periofi at room temperature. 17 Pzncmr loss 1 45 B^g. C 5S C 3 DC Deo. i (V me k®) (KM kfc) (Days) Fersa 1 *> i ' 1 4 § S 2 4 3 Xw wiesioa 3"? 7S - 57 » - 300 I M3S0i#j 2 „ $ to * S » *5 1.4 5 10 »6 ■» 3 0 tc •>! O-JO I 9.® 3.4 D.S8 JO. 3 3.7 3. 4 - SCI gailt 4.8 3.3 8. 0 6. 4 6.4 - «* - j sacii, 0 - 1,4 - 0 - 1.2 12. & ia,?D,|, Salt 0 3.0 3.0 - 3.7 5.0 - - Exarrple VIII Testing of Pnysical /^mixtures Physical admixtures -sere made vp of crystalline sulfuric acid salt with (a) tri sodium orthopho^ihate, (b) sodium 15 bicarbonate, (c) L(-s-) lysine, and (d) L(->) arginine0 "Die bases ware added in proportions to provide pH's on dilution of the admixture with water to a zwitterion activity of 250 mg/ml (as determined by HHLC assay) as follows: trisodium orthcphosp'nate (to provide a pH of 6,0); sodiusn bicarbonate (to provide a pH of 20 5.0); L(-f) lysine (to provide a pH of 5.0); L(-0 arginine (to i 8 provide a pH of 6. OS. Injectable compositions were made wp by reconstituting with sterile water to a zwitterion activity of 250 tag/ml «® determined by HPLC assay. There were no solubility problems. Injection© C100 ag/kgi were serried out intramuscularly on rabbits with pain within acceptable thresholds. The least paiift, was with the arginine containing composition.

Similar results of gocxS solubility and acceptable pain on intramuscular injection are obtained on use of the other salts Sserein in the physical admixtures with the above bases„ Figure 1 is the infra red absorption spectrum of the crystalline sulfate salt prepared as described in JQcsmples % or 21 pelletized in the crystalline form with potassium bromide.

The X-ray powder fliff raction pattern of the crystalline sulfate salt of 7-[c-(2-asninothia2ol-4-ylS-s-izj-methoscyimino-&cetamido]-3-I (1-snethyl-l-pyrrolidinio) -methyl]-3-cephem-4-carboxylate prepared as described in Example I or II was determined with a Stigaku Powder Piffractometer using a copper target X-ray tube, a nickel filter, and the sample contained in a glass dish. The scan rate was 2 «Seg»/ain. over the range from 5 deg. to 40 deg. and a chart %?as saechanically recorded to show the angles of maximum diffraction. From this the (d) spacings and relative intensities ^ere calculated. They are listed below. 1 9 o © d spacing I'JU ■ .1/1 US 9.20 . 100 ■v 6.80 SO 5.50 28 5 5.09 22 €.50 3® 4.41 44 4.19 63 3.78 30 SO 3.64 41 3.39 25 3.31 31 3.15 47 V> 2 © EXAMPLE IX PREPARATION OF THE SESOUIPHOSPHATE SALT The zwitterion, 0.70 g», is dissolved with rapid stirring in from 2.2 to 2.4 ml. of 85% phosphoric acid (2.1 to 2.2 molar equivalents) which has been diluted 1:10 (v/v) with water. The solution is clarified by filtration through a 0.22-0.45 micron pore-sise membrane filter. From 5 to 7 parts by volume (15-20 ml) of methanol is added to the filtrate with rapid stirring during a 30 to 50 min. period. Crystals form during this operation, and rapid stirring is continued for 1.5 to 2 hours. The crystalline product is recovered by vacuum filtration. The product is washed on the filter first with 6 to 8 ml of Is 1 (v/v) methanols acetone taking care to maintain a tightly packed filter cake, and then with acetone. The product is dried in vacuo at 50° C for 2 hours; typical yield 0.7 to 0.75 g.

Infrared Interpretation (See Figure 2) (IR, KBr pellet) Peak Position (cm-1) Functional Group 2800-3400 1780 NH , f carhosey 1 OH lactam C=0 1580 Carboxyl C=0 1550 Amide C=0 1S30 C=H,C=C 1550 Amide OH 980,1040 PO Behavior On Heating ^ An exotherm is shown at 171 „8°C in the differential scanning calorimeter tracing.

V X-Ray Diffraction Pattern 5 The X-ray powder diffraction pattern of the foregoing sesquiphosphate salt was measured with a Rigaku Powder Difftactometer in the same fashion as described above with respect to sulfate salt with the following results. d I/I 11.04 _ 32 9.2 — 16 7.89 — 24 7.02 — 42 6.7 - 32 5.5 — 26 4.64 - 100 4. 456 — 53 4.3 — 58 3.88 - 26 3.75 — 89 3.56 — 21 3.33 - 20 3.05 - ] 6 MjflR Interpretation (~H 90 MHz HMRt, D,0 solution) v m Chemical Shift (ppm 6 vs. TSP) Description Integral Assignment 2.0-2.4 Multiplet si 14CH2f, 149CH2 3.04 Singlet 3 12CH3 3 o 3-3»6 Multiplet 5 2CH, 13CH2„ 13'CH2 3„94 Doublet 1 2CH 4»12 Singlet • 3 20CH3 4.12 Doublet 1 11CH 4.8 Doublet 1 11CH 5.42 Doublet 1 SCH 5.88 Doublet 1 7CH 7.21 Singlet 1 18CH c Stability 2 3 10 Time-Temperature % Loss 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 - 1.4 4 weeks; 56°C - 0 1 week; 4 5°C - ■ 0 2 weeks; 45°C - 1.4 4 weeks; 45°C - 0.7 8 weeks; 4 5°C - 1.6 1 month; 37°C - 2.5 Elemental Analysis (percent by weight) 15 Found Dry Basis Theory (Sesauiphosphate) 20 N h3P°< 35.44 a S 12.38 2„29* 23.06 36.3 4.41 13.2 23.6 36.4 • 4.7 13.4 monohydrate =2.8% H-O 23.6 "Karl Fischer Method 24.

Claims (1)

1. CLAIMS % j. A tenperature stable crystalline salt of 7-J®-C2-•minothiasol-4-yl)-e" f II -anethoxyiminoacetamido]-3-1 Cl-soethyl-l~ gyrrolidinio) -snethylJ-3~cephem-4~carboxylate selected from 5 the sulfuric, di-nitr.ic, mono-hydrochloric, and di-hydrochloric acid addition salts, and orthophosphoric acid addition salts containing 1.5-2 siolar equivalents of H^PO^, or a solvate thereof. 2. 'The crystalline salt of Claim 1 selected from 10 the sulfuric, mono-hydrochloric, di-hydrochloric and orthophosphoric acid addition salts, or a solvate thereof. 3. A physical admixture of the salt of Claim I or 2 with a pharmaceutically acceptable non-toxic organic or inorganic base in proportions to provide a pH of about 3.5 to 7 on 15 dilution of the admixture with Mater to injectable concentration. 4. h physical admixture as recited in Claim 3 wherein the salt and th® base as© present in proportions to provide a pS of about 4 to 6 on dilution of the admixture with water to injectable concentration. 20 5. .ft physical admixture as recited in Claim 3 or 4 wherein the base is M+! lysine. 6. ft, physical admixture as recited in Claim 3 or 4 wherein the base is LC+S arginine. 7® A nethod for jaaking the sulfuric acid addition salt of Claim 1 eotcpri«i!»g the steps of |'®| fforsing an aqueous admixture of (i) at least 1 nolar equivalent oi naslfuric acid and Jii) svitterion corresponding to said salt, |b| causing crystallisation of the sulfuric acid addition •Alt to occur, provided that %»°hen the svitterion is present in the aflaixture at m concentration of less than 2S ag/sal the crystallisation is carried out in the presence of an organic •olvent, and I'd! isolating crystalline sulfuric acid addition salt. e. The method as recited in Claim 7 wherein step (b) is carried out is an aqueous a«diu:m free of organic solvent™ $. The aethod as recited in Claia 7 or 8 wherein the zwitterion is used in step (a) in an amount so as to be present in the admixture at a concentration less than SOO sg/al 10. The aethod a® recited in any aie of Claims 7{, 8 cr 9 wherein the amount of svitterion employed in step (a) is such that it is present in the ftdaiactwre at a concentration greater than 25 mg/ml. 2 6 11. The method as recited in Claim 8 wherein the zwitterion is used in step (a) in an amount so as to be present in the admixture at a concentration ranging from about 100 mg/ml to 200 mg/ml, 12. The crystalline sulfuric acid addition salt of Claim 1. 13. Crystalline 7- [ a - ( 2-arrvinothiazol -4-yl )-a-(Z)-methoxyiminoacetamido]-3- [ (1-nethyl-l-pyrroliainio)- . methyl]-3-cephem-4-carbaxylate sulfate salt having the following X-ray powder diffraction pattern: ° o d ssacina (A) I/I (%) 9.20 100 6.86 50 5.5 0 28 5. 09 22 4.50 38 4 . 41 44 4.19 63 3.78 38 3.64 44 3. 39 25 3.31 31 3.15 47 37 14. The crystalline orthophasphoric acid addition salt of Claim or a hydrate thereof. 15. Crystalline 7- [a- (2-asi.ittothiaaol-4-yl) -a- (E)-methoscyiminoacefcamido3 -3- [ (1-methylpyrrolidinio) -methyl]-3-cephem~4-earboxylate phosphate having the following x-ray powder diffraction pattern: d 1/1° 11.04 32 16 24 42 32 26 100 53 58 26 89 21 26 16 10 15 9.2 7.89 7.02 6.7 5.5 4.6 4 4.456 4 „ 3 20 3.88 3.75 3~5S 3.31 3.05 2 8 16. A method for making a salt or solvate as claimed In claim 1, substantially as described in any of Examples I to VI or Example XX. 17. A salt or solvate as claimed in Claim 1, made by a method as claimed in any of Claims 7 to 11f or Claim 16. 18a, A composition comprising a salt or solvate as claimed in Claim 17 in physical admixture with a pharmaceutically acceptable normally solid non-toxic organic or inorganic base. F. R. KELLY & CO., AGENTS FOR THE APPLICANTS.