WO2012095438A1

WO2012095438A1 - Particles and suspensions of cephalosporin antibiotics

Info

Publication number: WO2012095438A1
Application number: PCT/EP2012/050335
Authority: WO
Inventors: Hans Peter Niedermann; Heiko Bothe
Original assignee: Intervet International B.V.
Priority date: 2011-01-12
Filing date: 2012-01-11
Publication date: 2012-07-19

Abstract

Disclosed is a method of making particles of a cephalosporin antibiotic, particularly cefquinome, wherein use is made of diafiltration. The diafiltration can be with anti- solvent, in which case a precipitate is obtained of particles as such. The diafiltration can also be with a pharmaceutically acceptable suspension medium. In that case several process steps of isolating, drying, transporting of particles can be avoided, because the suspension resulting from the synthesis of the particles is directly turned into a final drug product formulation.

Description

PARTICLES AND SUSPENSIONS OF CEPHALOSPORIN ANTIBIOTICS

Field of the Invention The invention pertains to a method of making particles, particularly crystalline particles, of cefquinome or other cephalosporin antibiotics, and to a method of making

suspensions of such particles. The invention also pertains to the use of diafiltration in making particles and suspensions of cephalosporin antibiotics, particularly of

cefquinome sulfate.

Background of the Invention

Cefquinome is a cephalosporin antibiotic used for humans and animals, inter alia, to treat bovine respiratory disease, Pasteurella infections in pigs, and many other applications where a high antibacterial activity is desired. Cefquinome generally comes as an acid addition salt, preferably a sulfate. The cefquinome sulfate has a chemical structure of the following formula:

The chemical name for this structure is 1-[[6R,7R)-7-[2-(2-amino-4- thiazolyl)glyoxylamido]-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0] oct-2-en-3-yl]methyl]- 5,6,7,8-tetrahydroquinolinium hydroxide, inner salt 7²-(Z)-(0-methyloxime), sulfate.

The synthesis of cefquinome sulfate and other acid addition salts is known, e.g., from EP 280 157 or US 2006/0100424. Existing processes for making sterile cefquinome sulfate particles comprise the steps of providing a suspension of cefquinome sulfate in water; adding alkaline (typically NaOH) to the sulfate, followed by an organic solvent, (typically acetone), resulting in the precipitation of the corresponding sulfate salt (Na₂S0₄), and the retaining of a solution of cefquinome free base (which in fact is a betaine); conducting sterile filtration of the betaine solution; and adding sulfuric acid so as to produce sterile cefquinome sulfate particles or adding any other acids to produce any corresponding sterile cefquinome salt particles.

The drug is presented inter alia as an injection preparation, typically in the form of suspensions, e.g. in ethyl oleate or in the type of esters of saturated coconut and palm kernel oil-derived caprylic and capric fatty acids and glycerine or propylene glycol, known by the trade name of Miglyol. As described in WO 03/063877 the use of an antibacterial agent in the form of small particles of a narrow particle size range is advantageous compared to coarser material. A practically feasible size distribution of such small particles is, e.g.,

d(50) ≤ 7 μηι

d(90) ≤ 15 m

d(100)≤ 50 μΓΠ

Hereto the aforementioned sterile cefquinome sulphate particles, prepared according to EP 280157 or US 2006/0100424, are too coarse and have to be reduced in size, by high energy particle size reduction methods, typically by milling or micronizing.

Although cefquinome generally yields stable products, a desire exists to even further improve cefquinome particles in terms of physical stability and to provide cefquinome suspensions that are stable, particularly with reference to lower vulnerability for sedimentation. Moreover, the foregoing should preferably be realized on the basis of cefquinome particles that satisfy the aforementioned size distribution.

This desire is addressed in co-pending patent application PCT/EP2010/060376. Therein particles of a cefquinome acid addition salt, preferably cefquinome sulfate, are prepared by first making a suspension of cefquinome acid addition salt, and precipitating the salt from said suspension.

Also for other cephalosporin antibiotics, it is generally desired to provide the drug in the form of particles, particularly crystalline particles, of a relatively small size, and of a desired size distribution. The cephalosporin antibiotics particles are ultimately formulated into a drug product, preferably in the form of a suspension of said particles.

The formation of the desired particle sizes and the desire of obtaining a stable suspension in a pharmaceutically acceptable liquid medium, presents several technical challenges. One is that obtaining particles, notably crystalline particles, requires precipitation from a solution or suspension in a controlled way. To this end, e.g. in the case of cefquinome sulfate, acetone is added to water, or to a mixture of water and acetone, so as to force precipitation. This brings about a relatively high volume of solvent (or, rather, the anti-solvent acetone), which moves the production of

commercially viable batch sizes in an uneconomical direction.

It would therefore be desired to provide a method by which particles, notably crystalline particles, of the desired particle size can be obtained on the basis of a substantially reduced volume of solvent.

Further, in the existing processes, the final drug product is a suspension, in a pharmaceutically acceptable liquid medium such as ethyl oleate, of the crystalline particles as produced in the desired particle size at an earlier stage of the process. The overall process therefore requires not only the controlled precipitation of the particles, but also isolation, drying, packaging, transportation, and formulation. It would be desired to be able to dispense with one or more of the steps in between the formation of the particles (viz. in a suspension) and the production of the final formulation (also a suspension).

Summary of the Invention In order to better address one or more of the foregoing desires, the invention, in one aspect, presents a method of making a composition comprising particles of a cephalosporin antibiotic, wherein the cephalosporin is initially produced in the form of a suspension in a first liquid medium, wherein the suspension is subjected to cross-flow filtration against a further liquid medium, said further liquid medium being selected from media behaving as an anti-solvent for the cephalosporin and pharmaceutically acceptable media capable of retaining the cephalosporin as a suspension.

In another aspect, the invention provides the use of a diafiltration method in the production of particles, particularly crystalline particles of a cephalosporin antibiotic. In yet another aspect, the invention provides a method of isolating cephalosporin antibiotics particles from a solution or suspension in a liquid medium by the addition of an anti-solvent for the antibiotic, wherein anti-solvent is added by means of diafiltration.

In a further aspect, the invention provides a method of making a pharmaceutical formulation comprising a suspension of a cephalosporin antibiotic in a pharmaceutically acceptable liquid medium, wherein the formulation is made from a crystalline suspension in a processing liquid medium, and wherein said pharmaceutically acceptable liquid medium is substituted for said processing liquid medium by means of diafiltration.

In a still further aspect, the invention pertains to the use of liquid medium exchange by diafiltration, in the preparation of suspensions of cephalosporin antibiotics.

Brief description of the drawings

Fig. 1 shows a schematic set-up of an embodiment of the invention in which diafiltration is used in the generation of cephalosporin particles.

Fig. 2 shows a schematic set-up of an embodiment of the invention in which diafiltration is used in directly converting a sterile cephalosporin suspension as obtained in a particle generation step, into a drug product (suspension formulation in a pharmaceutically acceptable liquid medium).

Detailed description of the Invention

In a broad sense, the invention is based on the judicious recognition to use diafiltration as a method for exchange of liquid media, in the preparation of suspensions of cephalosporin antibiotics. According to the invention, the method can be applied either in the working-up of the cephalosporin from its synthesis so as to obtain the

cephalosporin in particulate form, or in the direct production of a drug product comprising the cephalosporin, wherein the step of first isolating particles can be bypassed.

The present invention will further be described with respect to particular embodiments and with reference to certain drawings but the invention is not limited thereto but only by the claims. Any reference signs in the claims shall not be construed as limiting the scope. The drawings described are only schematic and are non-limiting. In the drawings, the size of some of the elements may be exaggerated and not drawn on scale for illustrative purposes. Where the term "comprising" is used in the present description and claims, it does not exclude other elements or steps. Where an indefinite or definite article is used when referring to a singular noun e.g. "a" or "an", "the", this includes a plural of that noun unless something else is specifically stated.

The invention pertains to cephalosporins. These are a class of beta-lactam antibiotics generally being considered derivatives of 7-aminocephalosporanic acid. Many cephalosporins are on the market. The non-proprietary names generally start with "cef" (and sometimes ceph). Preferred cephalosporins include cefclidine, cefepime, cefluprenam, cefoselis, cefozopran, cefpirome, ceftiofur, cephapirin, cefotaxime, cefadroxil, cephalexin, cefovectin, cefazolin, cephalothin, and cefquinome. More preferably the cephalosporin is cefquinome, and most preferably a cefquinome acid addition salt.

In order to prepare an acid addition salt of cefquinome, a suitable acid is added to the betaine solution. These acids can be organic or inorganic, monobasic or dibasic acids, with mineral acids being preferred.

Suitable acids include, e.g., HCI, HBr, HI, HF, H₂N0₃, HCI0_4, HSCN, aliphatic mono-, di- or tricarboxylic acids, for example acetic acid, trifluoro acetic acid, trichloro acetic acid, or a preferred physiologically acceptable acid, such as, for example, the maleic acid, so as to provide a salt having the monomaleate anion HOOCCH=CHCOO^" .Another suitable organic acid is naphthoic acid. Preferred cefquinome acid addition salts include cefquinome dihydrochloride, cefquinome dihydroiodide, cefquinome sulfate, cefquinome-6-hydroxy naphthoate, cefquinome-naphthoate, cefquinome 2,4 dihydroxy benzoate. The most preferred salt is the cefquinome sulfate.

Cephalosporins generally come as suspensions in a pharmaceutically acceptable liquid medium, particularly in the form of a suspension in an oily base, being a vegetable oil or a mineral oil or mixtures thereof, e.g. peanut oil, castor oil, ethyl oleate, liquid paraffin, MCT oil (see below) or, Miglyol® (see below). The base preferably comprises a pharmaceutical acceptable low viscosity oily medium, such as medium chain triglyceride or a mixture of medium chain triglycerides. Medium chain triglycerides (MCT oil) have fatty acid chains of 6 - 12 carbon atoms and for the medically refined grades of MCT oil each chain has 8 - 10 carbon atoms. The MCT oil may comprise either triglycerides of the C8-C10 fatty acids, or propylene glycol diesters of these fatty acids or a mixture of both triglycerides and propylene glycol diesters. Preferably these C8 -C10 fatty acids are fully saturated, such as n-caprylic and n-capric acids. These are conveniently prepared by the commercial fractionating of naturally occurring vegetable (e.g. coconut) oil to give mainly C8-10 fatty acids followed by esterification of these acids with a chosen alcohol. Fractionated vegetable oil having the desired composition is commercially available. Proprietary examples of such oils are Miglyol® 812 as capric/caprylic triglycerides and Miglyol® 840 as propylene glycol dicaprylate/caprate.

Most preferred is Miglyol® grade 812. In existing processes, the suspension is made by first making solid particles, preferably crystalline particles, which are isolated, dried, micronized, packaged, and transported and, ultimately, rendered into a

suspension formulation. The micronization step can be avoided, via the process of copending application no PCT/EP2010/060376. In this process, the step of particle generation is conducted in such a way as to obtain a suspension of crystalline particles of the desired particle-size distribution.

Whilst the step of particle generation will result in a suspension (particularly a sterile suspension), the liquid medium of the suspension will have to be changed in favor of an anti-solvent in order to have the particles precipitate (e.g. acetone in the case of cefquinome sulfate). This step, and the relatively high amounts of anti-solvent needed, considerably reduces the batch size at which the cephalosporin particles can be produced. The present inventors have found a way of adding the anti-solvent by means of diafiltration (cross-flow filtration). An advantage hereof is that a much lower amount of anti-solvent is required, and the particles can be produced at desired commercial batch sizes.

Thus, in one embodiment, the invention is a method of making a composition comprising particles of a cephalosporin antibiotic, wherein the cephalosporin is initially produced in the form of a suspension in a first liquid medium, wherein the suspension is subjected to cross-flow filtration against a further liquid medium, and wherein said further liquid medium is selected from media behaving as an anti-solvent for the cephalosporin.

Anti-solvents are known, and when a cephalosporin is given, it is within the ambit of the average skills of the artisan to select suitable anti-solvents. In the case of cefquinome or cefquinome sulfate or other acid addition salt, a suitable anti-solvent is acetone or a mixture of water and acetone wherein acetone is the major component.

Other suitable anti-solvents are within the ambit of the artisan's skills. In carrying out the diafiltration of the invention, the composition of the further liquid medium can be changed as desired during the process, e.g. by gradually increasing the amount of acetone in the case of using a mixture of water and acetone as an anti-solvent. In this embodiment, the composition comprising particles of a cephalosporin antibiotic in fact comprises the isolated (precipitated) particles as such.

In a preferred embodiment, the further liquid medium is selected from

pharmaceutically acceptable media capable of retaining the cephalosporin as a suspension. As a result, in this embodiment, the invention puts to use the

aforementioned diafiltration in a different way. Rather than using the technique in the existing process, viz. at the step of isolation of particles, the preferred embodiment uses the diafiltration of the suspension of cephalosporin particles to add the eventual liquid medium of the drug product, i.e. the aforementioned oily liquid.

In this embodiment, the composition comprising particles of a cephalosporin antibiotic is a pharmaceutical composition in the form of a suspension of the

cephalosporin antibiotic in a pharmaceutically acceptable liquid medium. This leads to considerable advantages, in that all of the process steps associated with isolating cephalosporin particles can be dispensed with. Thus, rather than involving the steps of isolation, drying, particle size adjustment (micronization), packaging, transportation, and reformulation as a suspension, this embodiment of the invention allows a direct step from a sterile crystal suspension towards a final drug product suspension.

Depending on the pharmaceutically acceptable liquid medium used in the drug product suspension and the liquid medium in which the cephalosporin crystal suspension is produced, these liquid media may or may not be sufficiently miscible with each other so as to conduct the diafiltration. In the latter case it is preferred to conduct an additional diafiltration with an intermediate (second) liquid medium, that is miscible with both the first liquid medium (in which the crystals are produced) and the further liquid medium (the pharmaceutically acceptable liquid medium in the drug product). In the table below, guidance is given on several example liquids used. The possible liquids are not limited to the ones mentioned herein, and the person skilled in the art will be able to easily determine whether or not liquids are miscible, so as to decide whether or not the diafiltration can be conducted directly, or via an intermediate liquid.

Table

Miscibility of various liquids

acetone (0,32) Miglyol® (30) - yes acetone (0,32) ethyloleate (6.5) - yes acetone (0,32) liquid paraffin (110-230) - no iglyol® (30) liquid paraffin (110-230) - yes iglyol® (30) ethyloleate (6.5) - yes ethyloleate (6.5) liquid paraffin (110-230) - yes

Miglyol® (30) 32% acetone (0,32) 40% liquid paraffin (110-230) 22% yes

Miglyol® (30) 35% acetone (0,32) 50% liquid paraffin (110-230) 15% yes ethyloleate (6.5) 25% acetone (0,32) 25% liquid paraffin (110-230) 50% yes ethyloleate (6.5) 22% acetone (0,32) 30% liquid paraffin (110-230) 48% yes

In the invention, a cephalosporin suspension is subjected to cross-flow filtration against a further liquid medium, said further liquid medium being selected from media behaving as an anti-solvent for the cephalosporin and pharmaceutically acceptable media capable of retaining the cephalosporin as a suspension. It will be understood that the pharmaceutically acceptable liquid medium will generally not be a solvent for the cephalosporin, as it retains a suspension thereof. It will also be understood, in view of the above embodiment to make use of an intermediate liquid medium, that the invention encompasses the indirect exchange of the liquid medium in the initially obtained suspension with the final pharmaceutically acceptable liquid medium.

Cross-flow filtration (also referred to as diafiltration) is a known technique, normally used for entirely different purposes. Typical uses include waste-water treatment, the production of purified water or salt water purification. A typical use in pharmaceutical industry is for the enrichment of a fermentation slurry. A further typical use in health care environment is the well known dialysis of human blood. As termed in the "Dictionary of Filtration and Separation," diafiltration is a membrane based separation that is used to reduce, remove or exchange salts and other small molecule contaminant from a process liquid or dispersion. The process essentially involves passing a feed flow across a membrane filter on one side, and passing a clean liquid across the membrane filter on the other side, in the opposite direction. This type of filtration is typically selected for feeds containing a high proportion of small particle size solids (where the permeate is of most value) because solid material can quickly block (blind) the filter surface with dead-end filtration. In batch diafiltration, the process fluid is typically diluted by a factor of two using "clean" liquid, brought back to the original concentration by filtration, and the whole process repeated several times to achieve the required concentration contaminant. In continuous diafiltration the "clean" liquid is added at the same rate as the permeate flow. Cross-flow filtration refers to the fact that the majority of the feed flow travels tangentially across the surface of a filter, rather than into the filter. The principle advantage of this is that the filter cake (which can blind the filter) is substantially washed away during the filtration process, increasing the length of time that a filter unit can be operational. It can be a continuous process, unlike batch-wise dead-end filtration.

The manner in which the present invention puts to use the technique of diafiltration deviates from the regular uses thereof in several respects. In one respect, the technique is used in the invention to precipitate solids (viz. cephalosporin particles), which in classical filtration would mean keeping them as a retentate. In another respect, the technique is used to transfer cephalosporin from one suspension to another, which is entirely incomparable with normal (filtration-type) used of diafiltration.

In the diafiltration process of the invention, the filter will preferably be a ceramic membrane. Such filters are known, and are made of, e.g., silicon, silicon nitride, silicon oxynitride, silicon carbide, silicides, alumina, zirconium oxide, magnesium oxide, chromium oxide, titanium oxide, titanium oxynitride, titanium nitride, yttrium barium copper oxides, or as composite membrane, made of polymer membranes based on polyvinylalcohols, polyimides, polyacrylamides, polysiloxanes, polydimethylsiloxanes, ethylene-propylene diene, polynorbonene, polyoctenamer, polyurethane, butadiene, nitrile butadiene rubber, polyethylene, polypropylene, polyvinylidene difluoride, derivatives and mixtures thereof, on an inorganic (e.g. ceramic or zeolithe) surface layer The filter membranes can have a variety of shapes, e.g. disk-type or hollow tube- type.

It will be understood that the membrane has pores, which presents the skilled person with the choice of a pore size. The lower limit in the present invention is 1 nm diameter. The upper limit is 10 μηι. The diafiltration process can be conducted in equipment known in the art.

Typically, the equipment will comprise a filtration unit (1) comprising a tubular or disc diafiltration membrane (2), an inlet (3) for the flow (a) of liquid to be subjected to diafiltration, an outlet (4) for a flow of retentate (b) and an outlet (5) for a flow of permeate (c); said filtration unit comprises fluid connections (flows a and c above) with a mixing unit (6), comprising a vessel (7) provided with an agitator (8), an inlet (9) for a flow of anti-solvent (d), in the figure indicated with reference to acetone, an inlet (10) for the flow of permeate (c), and an outlet (11) for a flow of liquid (a), i.e. a suspension, from the vessel (7) to the filtration unit (1), driven by a pump unit (12).

In Fig. 1 the equipment is presented for use in generating cefquinome (or other cephalosporin antibiotic) crystalline particles, with reference to the use of acetone as the anti-solvent flow (d) and comprising a valve (13) allowing the flow of liquid (a) to either be led to a separation unit (not shown) so as to obtain solid particles, or to the filtration unit (1) for further diafiltration.

In Fig. 2 the aforementioned equipment is presented for use in generating a cefquinome (or other cephalosporin antibiotic) drug product (suspension), with reference to the use of a choice of carrier liquids as the anti-solvent flow (d) and wherein the valve (13) allows the flow of liquid (a) to either be led to a drug product filling line (not shown), or to the filtration unit (1) for further diafiltration.

The judicious choice of this method leads to advantages that are particularly enjoyed when making cephalosporin drug products. Therefore the invention, in one aspect, also relates to the use of a diafiltration method in the production of particles of a cephalosporin antibiotic.

The reason that the method provides such advantages is, inter alia, that the production of cephalosporin drug products requires a step of adjusting particle size, and requires a step of formulating the drug product as a suspension in a pharmaceutically acceptable liquid. The former step generally requires the use of liquids, such as acetone, that are not pharmaceutically acceptable. The use of a diafiltration method according to the invention, presents a much more economical process, with a lower risk of losing sterility in intermediate process steps, than in classical processes where cephalosporin particles first need to be isolated, and later reformulated.

In this respect, in a further embodiment, the invention also pertains to the use of liquid medium exchange by diafiltration, in the preparation of suspensions of

cephalosporin antibiotics. In another aspect, the invention relates to a method of making a pharmaceutical formulation comprising a suspension of a cephalosporin antibiotic in a pharmaceutically acceptable liquid medium, wherein the formulation is made from a crystalline suspension in a processing liquid medium, and wherein said pharmaceutically acceptable liquid medium is substituted for said processing liquid medium by means of diafiltration.

In yet another embodiment, the invention also pertains to a method of isolating cephalosporin antibiotics particles from a solution or suspension in a liquid medium (particularly a sterile suspension), by the addition of an anti-solvent for the antibiotic, wherein anti-solvent is added by means of diafiltration. The judicious insight to transport existing diafiltration technology to the entirely different field of the preparation of particles of a cephalosporin antibiotic brings about specific advantages. Particularly, since the anti-solvent generally is not a pharmaceutically acceptable liquid, it is desired to use as low an amount thereof as possible.

Moreover, in the case of cephalosporin antibiotics the production of a suspension of particles, notably of crystalline cephalosporin, it is important that sterility be

maintained. This means that also the anti-solvent needs to be sterile, and needs to be used in a sterile equipment area. This adds to the technical complexity, and costs, when relatively high amounts of anti-solvent need to be used.

In summary, the invention presents a method of making particles of a

cephalosporin antibiotic, particularly cefquinome, wherein use is made of diafiltration. The diafiltration can be with anti-solvent, in which case a precipitate is obtained of particles as such. The diafiltration can also be with a pharmaceutically acceptable suspension medium. In that case several process steps of isolating, drying, transporting of particles can be avoided, because the suspension resulting from the synthesis of the particles is directly turned into a final drug product formulation.

The invention is further illustrated with reference to the following, non-limiting examples, and the accompanying drawings.

Examples

General set up of the experiments

The diafiltration trials were done based on a cefquinome sulfate suspension with contents of 7,5-8% cefquinome sulfate in a mixture of acetone and water with a ratio of 2: 1. as described in (PCT/EP2010/060376). The suspension was given to a jacketed and stirrable reservoir. By switch on the pump the loop was started. The desired temperature could be controlled by cooling the jacketed vessel. According to the valve position the permeate was recirculated into the reservoir or discharged during the concentration step and diafiltration step.

The suspension was concentrated to the desired value (normally to two third or the half of the initial concentration). Afterwards acetone was added with the same rate as the permeate left the loop, so the volume kept constant. The amount of the added acetone determined the diafiltration factor D (normally D=2.2) and finally the residual water content in the final suspension. After the diafiltration was finished, the suspension was filtered and dried.

In cases of change of suspending agents (anti-solvents) the acetonic suspensions were diafiltrated with the appropriate agent (anti-solvent) producing the corresponding stem suspensions.

Example 1

The study was done on a ceramic membrane with a porosity of 150 kD. 0.7 L of a crystal suspension was concentrated to 0.35 L and then diafiltrated with 0.8 L pure acetone at a steady volume. During this process step the water content decreases to 7.5%. The success of the diafiltration was tested with filterability tests on a pressure filter. Therefore a part of the original suspension was diluted with acetone to an acetone/:water ratio of 29: 1 v:v and filtered. This filtrate was turbid, a filtration time of 1 .4 min/g solid was observed. The diafiltrated suspension was also filtered, a clear filtrate and a filtration time of 0.6 min/g solid was observed.

Analytical results

water content of final filtrate: 7.5% (by density measurement)

specific filtration time to separate the crystals after diafiltration: 0.6 min/g

specific filtration time to separate the crystals after dilution to 29: 1 v:v acetone/water: 1.4 min/g Example 2

Example 2 was performed according to Example 1 with a diafiltration factor D=1.5, resulting in a water content of 10% for the final suspension and a filtration rate of 1.3 min/g.

Example 3

Example 3 was performed according to Example 1 with a diafiltration factor D=2.2, resulting in a water content of 5% for the final suspension and a filtration rate of 0.2 min/g.

Example 4

Example 4 was performed according to Example 1 with a diafiltration factor D=3.0, resulting in a water content of 2% for the final suspension and a filtration rate of 0.1 min/g.

Example 5: Scale up

Example 5 was performed according to Example 1 with a volume of 5 L, using a membrane with a surface of 0.2 m², ten times larger as for lab scale, and with a diafiltration factor D=2.5, resulting in a water content of 2% for the final suspension and a filtration rate of 0.1 min/g. The time needed for concentration and diafiltration was half the time as for the appropriate experiments, due to doubling the specific surface area per suspension volume. No scale up effect was detected. Example 6

Example 6 was performed according to Example 5 with a diafiltration factor D=2.5, resulting in a water content of 2.3% for the final suspension and a filtration rate of 0.1 min/g, but using a 50 kD ceramic membrane Example 7

Example 7 was performed according to example 1 , using ceramic discs with a porosity of 60 nm and an appropriate equipment. 6.1 L of the crystal suspension was diafiltrated with 25 L of acetone, resulting in a calc. water content of lower than 1 %. The final suspension was filtrated with a filtration rate of < 0.1 min/g. Example 8

Cefalonium-dihydrat was suspended in acetone: water 2: 1 , resulting in a suspension of 10% cefalonium-dihydrat related to water. The diafiltration process was carried out with the same filter equipment as described in example 7.

4.7 L of the suspension was diafiltrated with 23 L of acetone, resulting in a water content < 1 %. The final suspension was concentrated to a volume of 4 L and filtrated with a filtration rate of < 0.1 min/g. Solvent Change to Formulation Suspensions

Example 9:

The initial suspension for this experiment had a content of 7.5-8% Cefquinome sulfate in a mixture of acetone and water with a ratio of 2: 1.

The trial was performed by

1. concentrating at 20°C of the initial suspension with X= 1.6,

2. by diafiltration at 20°C with acetone with D=2.5 (residual water content less than 3%),

3. diafiltration with ethyloleate at 30°C with D=5.3 (residual acetone content less than 1 %)

Membrane type: porosity 150 kD; area 220 cm²

The resulting ethyloleate suspension had an acetone content < 0.2%, water content < 0.4 % and a cefquinome sulphate content of 8.6%.

Example 10:

Diafiltration with ethyloleate and liquid paraffin as solutizing agents

Example 10 was performed according to Example 9 with D=2.0 for diafiltration with ethyloleate at 30°C, followed by diafiltration with liquid paraffin at 30°C with D=5.0, resulting in a paraffin suspension with a calculated residual acetone content < 0.15% and a calculated residual ethyloleate content < 1%.

Example 11

Diafiltration with Miglvol^® and liquid paraffin as solutizing agents Example 11 was performed according to Example 9 with D=5.0 for diafiltration with Miglyol^® at 30°C, resulting in a Miglyol^® suspension with a calculated acetone content of <1 %. The Miglyol^® suspension was then diluted with acetone to come to a acetone content of 20% (D=2) and subsequently diafiltrated with liquid paraffin at 30°C with D=5.0, resulting in a paraffin suspension with a calculated residual acetone content < 0.15% and a calculated residual Miglyol^® content < 1 %.

Claims

1. A method of making a composition comprising particles of a

cephalosporin antibiotic, wherein the cephalosporin is initially produced in the form of a suspension in a first liquid medium, wherein the suspension is subjected to cross-flow filtration against a further liquid medium, said further liquid medium being selected from media behaving as an anti-solvent for the cephalosporin and pharmaceutically acceptable media capable of retaining the cephalosporin as a suspension.

2. A method according to claim 1 , wherein the further liquid medium is an anti-solvent for the cephalosporin, and the produced composition comprising particles of a cephalosporin antibiotic comprises the cephalosporin particles as such.

3. A method according to claim 1 , wherein the further liquid medium is a pharmaceutically acceptable medium capable of retaining the cephalosporin in suspension, and the produced composition comprising particles of a cephalosporin antibiotic is a suspension of the cephalosporin antibiotic in a pharmaceutically acceptable liquid medium.

4. A method according to claim 3, wherein the suspension in the first liquid medium is subjected to cross-flow filtration against a second liquid medium so as to obtain a suspension in the second liquid medium, said suspension in the second liquid medium being subjected to cross-flow filtration against the pharmaceutically acceptable liquid medium.

5. A method according to claim 4, wherein the second liquid medium is selected from the group consisting of ethyl oleate and Miglyol®, and the further liquid medium is liquid paraffin.

6. A method according to any one of the preceding claims, wherein the cephalosporin antibiotic is cefquinome or an acid addition salt thereof.

7. A method according to claim 6, wherein the acid addition salt is the sulfate.

8. A method according to any one of the claims 1 , 3, 6, and 7, wherein the further liquid medium is selected from the group consisting of ethyl oleate, MCT oil as defined in the description, and Miglyol®, as defined in the description.

9. A method according to claims 1 , 2, 6, and 7, wherein the further liquid medium is acetone or a mixture of acetone and water.

10. The use of a diafiltration method in the production of particles of a cephalosporin antibiotic.

1 1. A method of isolating cephalosporin antibiotics particles from a solution or suspension in a liquid medium by the addition of an anti-solvent for the antibiotic, wherein anti-solvent is added by means of diafiltration.

12. A method of making a pharmaceutical formulation comprising a suspension of a cephalosporin antibiotic in a pharmaceutically acceptable liquid medium, wherein the formulation is made from a crystalline suspension in a processing liquid medium, and wherein said pharmaceutically acceptable liquid medium is substituted for said processing liquid medium by means of diafiltration.

13. The use of liquid medium exchange by diafiltration, in the preparation of suspensions of cephalosporin antibiotics.