AU2001260918B2 - Carbohydrate medical solution and sulphite stabilisator in a multiple compartment container and use thereof - Google Patents

Description

WO 01/89478 PCT/SE01/01125 CARBOHYDRATE MEDICAL SOLUTION AND SULPHITE STABILISATOR IN A MULTIPLE COMPARTMENT CONTAINER AND USE THEREOF TECHNICAL FIELD The present invention relates to multiple compartment containers including sterile medical solutions, in which at least one solution contains carbohydrate compounds. The invention further relates to stabilising carbohydrates in a sterile medical solution.

BACKGROUND

Sterilisation of medical solutions such as, for example, peritoneal dialyses (PD) solutions, is commonly performed through the addition of energy, either in the form of radiation or heat. WO-A-9705852 discloses a multiple compartment container including sterile peritoneal dialyses solutions, which is heatsterilised in an autoclave.

In recent years scientists have become aware of the toxicity of decomposition compounds of carbohydrates in PD solutions. Wieslander et al., reported that all major brands of commercial PD solutions were toxic in contrast to PD solutions sterilised by filtration (Wieslander et al., 1991, Kidney Int, 40:77-79). The PD solutions were tested after dilution with cell growth media on cultured fibroblasts.

Furthermore, Wieslander et al. have reported that the glucose degradation products also affect the functional responses involved in host defence (Wieslander et al., 1995, Peritoneal Dialysis Int, 15 (suppl).

A patient on peritoneal dialysis (PD) uses between 8 and 20 litres of dialysis solution every day, depending on the treatment. This results in the consumption of 3-7 tons of solution with 1.5-4% glucose (50-175 kg pure glucose) every year.

(Wieslander, 1996, Nephrol Dial Transplant 11:958-959), which if the glucose undergoes decomposition also means a non-negligible amount of decomposition compounds. Furthermore, it is well known that some patients experience pain during inflow of the dialysis fluid. It has been speculated that the pain could be the result of glucose degradation (Henderson et al., 1985 Frontiers in peritoneal dialysis, ed.

O Winchester, New York: Field, Rich, 261-264) and that these degradation products Smediate basal cytotoxicity (Barile FA, 1994, Introduction to in vitro cytotoxicity.

SFlorida:CRC Press, 27-35). This means that they act upon fundamental life processes, which involve structures and functions common to all living cells such as membrane integrity, mitochondrial activity, or synthesis of proteins and DNA.

00oo These basal cell functions support organ specific cell functions. Thus, glucose degradation products capable of affecting basal cell activities are likely to interfere Swith specialised cell functions such as IL-1 13 release from mononuclear cells.

(Ni Glucose, an osmotic agent commonly used in PD solutions is known to degrade into carbonyls such as formaldehyde, acetaldehyde, metylglyoxal, 3deoxyglucosone and glyoxal.

Sulphite compounds have commonly been used as antioxidant in parenteral emergency drugs to prevent oxidation. The mechanism of decomposition of carbohydrates in PD solutions has appears however to have less to do with oxidation and sulphite is not intended to be used as an antioxidant in vitro in the present invention. Further, the anti-microbial or antioxidant compounds in parenteral emergency drugs are typically used in concentration which deliver 0.5 to 2 mg of sulphite per ml of undiluted drug injection (Smolinske S, 1992, Clinical toxicology 30:597-606). Such concentrations for preventing oxidation could not be used in PD solutions since they would administer too much sulphite to the patient resulting in adverse toxic effects.

SUMMARY OF THE INVENTION On the above background it is an object of the present invention to provide a multiple compartment container for sterile medical solutions of the kind referred to above in which decomposition of carbohydrates and/or the negative effects of the decomposition products are reduced. The multiple compartment container includes at least one sulphite compound in one or more of the compartments to stabilise decomposition of carbohydrates or to scavenge decomposition products formed during sterilisation and/or storage.

According to the first aspect of the present invention there is provided a multiple compartment container for sterile medical solutions, said solutions being present in two or more compartments, said container having breakable portions between two or more compartments, and at least one carbohydrate compound is 0 w present in one or more of said compartments, and at least one sulphite c compound is present in one or more of said compartments, wherein when the Sbreakable portions are broken the said solutions are mixed, N wherein said sulphite compound is selected from the group consisting of HS0 3

S

2 0 5 2 or S0 3 2 having a positive counter ion or mixtures thereof,, and oo wherein if said sulphite compound is a monosulphite compound, the total C0 amount of monosulphite compound in said container being equal to 0.05 1 \O mmol, per liter of the total amount of solution in said container, and if Ssaid sulphite compound is a disulphite compound, the total amount of 0 10 disulphite compound in said container being equal to 0.025 0.5 mmol, per liter of the total amount of solution in said container.

WO 01/89478 PCT/SE01/01125 3 The invention further relates to a medical solution wherein the solution contains at least one carbohydrate compound and at least one sulphite compound to stabilise decomposition of the carbohydrates or to scavenge decomposition products formed from the carbohydrates during sterilisation or storage of the medical solutions.

Additionally the invention relates to a method of stabilising a carbohydrate containing solution wherein the solution contains at least one sulphite compound to stabilise decomposition of carbohydrates or to scavenge decomposition products formed during sterilisation and/or storage.

Furthermore the invention relates to the use of a carbohydrate containing solution for the preparation of a multiple compartment container.

Finally, the invention relates to use of a carbohydrate containing solution for the preparation of a multiple compartment container for the treatment of a patient in need thereof.

BRIEF DESCRIPTION OF THE DRAWING In the following detailed portion of the present description, the invention will be explained in more detail with reference to an exemplary embodiment shown in the drawings, in which Figure 1 is a frontal view on a multiple compartment container according to an embodiment of the invention.

DETAILED DESCRIPTION OF THE INVENTION The invention is intended for use in treatments of diseases such as uremic disorder or kidney malfunctions, including for example treatments of diseases using peritoneal dialysis.

Definitions The term "multiple compartment container for a medical solution" is intended to mean any container including more than one compartment, particularly two or three but not limited to three, compartments. One example is a multiple compartment container used for peritoneal dialysis containing medical 0 5 solutions, which are sold under the brand Physioneal® and Gambrosol® trio.

The term "medical solution" is intended to mean any solution useful for oo medicinal purposes in which, a carbohydrate may be present and in which the Scarbohydrate undergoes decomposition during either the sterilisation procedure or storage resulting in disadvantageous decomposition products unfavourable for living cells. Decomposition products contemplated are for example products such as mono and dicarbonyl compounds, formaldehyde, acetaldehyde, methylglyoxal, 3-deoxyglucosone and glyoxal or the like. The storage conditions could be any conventional storage condition, such as room temperature for 2 years. One example of a medical solution is a solution, present in one or more of the compartments, used for peritoneal dialysis.

The term "final solution" is intended to mean a solution obtained by mixing one or more of the medical solutions in the container.

The term "peritoneal dialysis solution" is intended to mean a solution including an electrolyte, a buffer and an osmotically active compound, wherein the electrolyte comprises ions, such as sodium, potassium, calcium and magnesium; the buffer comprises components, such as acetate, lactate and bicarbonate; and the osmotic compound is a carbohydrate as defined hereinafter.

Examples of medical solutions for use as peritoneal dialysis solutions may be found in Wieslander et al., 1991, Kidney Int 40:77-79. The peritoneal dialysis solution could, prior to dialysis, be present in one or more compartments. In the case of multiple compartments the solutions are mixed prior to peritoneal dialysis.

The term "carbohydrate compound" is intended to mean sugars or sugars acids such as glucose, fructose, mannose, aldonic, alduronic, aldaric acids and their esters with saccharides or a polymer of glucose, fructose, mannose, aldonic, alduronic, aldaric acids and their esters with saccharides or a synthetic form of WO 01/89478 PCT/SE01/01125 glucose, fructose, mannose, aldonic, alduronic, aldaric acids and their esters with saccharides or derivatives and mixtures thereof.

The term "sulphite compound" is intended to mean a sulphite containing compound with the properties to reduce the content of decomposition products, by for example stabilising the solution, including preventing the generation of decomposition products, or scavenging already formed decomposition products, produced during sterilisation and/or storage of medical solutions containing carbohydrate compounds, as defined above.

Furthermore, the sulphite compound could be used as an antioxidant or to scavenge toxic or allergenic compounds in vivo, such as in the body fluids.

Examples of such toxic or allergenic compounds are metylglyoxal, 3deoxyglucosone and glyoxal. The effect of the sulphite compound can be measured according to the method "Analysis of glucose degradation products" mentioned under "Material and Methods" hereinafter. Examples of such sulphite compounds are any sulphite, having a positive counter ion, such as sodium, potassium, calcium, magnesium and ammonium, for example HSO3-, SO52 and S0 3 Examples of sulphite compounds to be used are NaHSO 3 Na 2

S

2 05 and Na 2

SO

3 or any other of sulphite compound or derivative thereof, natural or synthetic, or mixtures thereof.

The term "stabilising" is intended to mean preventing the generation of decomposition products, or scavenging already formed decomposition products, produced during sterilisation and/or storage of medical solutions containing carbohydrate compounds The term "carbohydrate decomposition products" is intended to mean products produced in a carbohydrate solution during any kind of sterilisation and/or during storage, which are products obtained from decomposition of carbohydrates, such as glucose and toxic to eucaryotic and procaryotic cells. Specifically contemplated are mono and dicarbonyl compounds, such as formaldehyde, acetaldehyde, methylglyoxal, 3-deoxyglucosone and glyoxal or the like. The toxicity can be measured according to the method "in vitro assay for cytotoxity" mentioned in "Materials and Methods hereinafter.

I

The term "sterilisation" is intended to mean any kind of sterilisation, such as radiation, pressure, heat, UV-radiation, radioactive radiation, sterile filtration, Sradiation using micro waves or other sterilisation methods. Furthermore the Ssterilisation can be performed using different approaches such as short 0 5 sterilisation time at a high temperature, sterilisation at low pH, sterilisation with high glucose concentration after removal of catalytic substances.

oo Multiple compartment containers employing a medical solution.

C The invention relates to multiple compartment containers for sterile medical 0solutions, particularly solutions for peritoneal dialysis wherein the medical c 10 solutions are present in one or more compartments. One or more of the compartments includes a carbohydrate and at least one sulphite compound in order to reduce the amount of the carbohydrate decomposition products produced during sterilisation and/or storage. Furthermore one or more of the compartments may include an electrolyte, a buffer and any other pharmaceutically acceptable additive or other component.

Additionally, the container includes at least two compartments, preferably three or more compartments, most preferably three. In at least one of the compartments there is provided a carbohydrate compound in solution and in at least one of the compartments there is provided a sulphite compound to reduce the formation or scavenge already produced decomposition products formed from carbohydrate. Furthermore the "sulphite compound" could be used as an antioxidant or to scavenge toxic or allergenic compounds in vivo, such as in the body fluids. Commonly used medical solutions either in single or multiple compartment container(s) for peritoneal dialysis preferably contain glucose in the final solution in a concentration in the range of 1.5 to 4,0 preferably substantially 1.5, 2.5 or 4 by weight (based on the final solution).

WO 01/89478 PCT/SE01/01125 7 Fig. 1 shows a preferred embodiment of the container, in this case a threecompartment bag The bag 1 is made from a continuous tube of a plastics material, which is sealed at both ends by sealing borders 2,3.

As shown in FIG. 1 each sealing border comprises several embossments 4 and apertures 5,6,7. The embossments 4 enhance the stability of the border 2. The aperture 5 in the upper border 2 is intended for hanging the bag during use and the apertures 6,7 in the lower border 3 are for fixation of the bag during the manufacturing operation.

The lower border 3 is also provided with an outlet tube 14, which connects compartment 9 with the consumer, for example a catheter ending in the abdominal cavity of a patient for peritoneal dialysis. Often tube 14 terminates in a luer connector (not shown in FIG. 1).

Furthermore, border 3 is provided with a filling tube 12, a medicament tube including a removable cap 16. When cap 16 is removed, tube 15 forms an entrance site for introducing any type of beneficial agent or medicament into compartnent 9 as desired, such as antibiotics.

The bag 1 is divided into three compartments 9,44,45 by welding seal lines 41,42,43. The upper compartments 44,45 divided by welding seal line 11 are of equal size and separated form the lower compartment 8 by two sloping welding lines 42,43. Thus there is formed a first upper compartment 44 and a second upper compartment 45, each being accessed via introduction tubes 46,47. The large lower compartment 9 comprises the electrolytes necessary for the solution to be formed (final solution), such as NaC1, MgC12, lactate etc., dissolved in water in a manner known per se.

The first compartment 44 comprises glucose solution having a concentration of about 30% and the second compartment 45 comprises a glucose solution having a concentration of about When breaking the breakable portion of connection tube 48, the contents of the first compartment 44 is mixed with the contents of the lower compartment 9 to form a peritoneal dialysis solution having a concentration of 1.5% of glucose. If the breakable portion of connection tube 49 is broken, the contents of compartment WO 01/89478 PCT/SE01/01125 8 is mixed with the contents of both compartment 9 thereby forming a dialysis solution having a concentration of about 2.5% of glucose. If both breakable portions of connection tubes 48,49 are broken, the contents of both compartments 44 and are mixed with the contents of compartment 9 thereby to form a dialysis solution having a concentration of about 4% of glucose. The above dialysis solutions formed by mixing at least one of the glucose containing compartments.

If the bag should be used as a nutritional solution, the large compartment 9 may comprise only NaC1 or any other suitable composition as used today but excluding glucose.

It is mentioned that the glucose can be exchanged with a glucose like component, such as glucose polymers, as an osmotic agent.

Furthermore, the sterile medical solutions comprising a carbohydrate, may include an electrolyte, a buffer such as lactate and any other pharmaceutically acceptable additive.

According to one embodiment of the invention the carbohydrate compound is separately provided in one or more compartments, the rest of the peritoneal dialysis solution compounds being provided in one or more the other compartments. The sulphite compound(s) may be present in one or more of said compartments or separately presented in one or more compartments. The sulphite compound(s) may be introduced in any of the carbohydrate or electrolyte solution compartments before or after sterilisation. However, some or all components of the medical solution(s) and the sulphite compound are mixed prior to peritoneal dialysis to obtain a final solution.

If a monosulphite compound such a bisulphate is used, it is preferably added to the carbohydrate and/or the electrolyte compartment in an amount to give a final solution within the range of 0.01 10 mM, preferably 0.05 1 mM, most preferably 0.05 0.5 mM. If a disulphite compound is used it is preferably added to the carbohydrate compartment in an amount to give a final solution within the range of 0.005 5 mM, preferably 0.025 0.5 mM, most preferably about 0.025 0.25 mM.

If a disulphite compound is used it is preferably added in an amount to give a final solution which is half of the concentration used for the corresponding Smonosulphite compound.

The pH of the solution in the carbohydrate compartment is preferably between pH 2.0-7.5, more preferably pH 2-5.5, even more preferably pH 3-4 and most preferably about pH 3.2. The pH of the final solution is preferably between oo 8.0, more preferably between 6.5 8.0, most preferably between 7.0 or absolutely most preferably 7.4.

in a preferred embodiment, the multiple compartment container containing the medical solution is sterilised. Any conventional methods and apparatus for sterilisation may be used, such as those mentioned under the definition of the term "sterilisation". Preferably the sterilisation is performed by heat treatment most preferable at about 121 C for 20 minutes (Ph. Eur.

(current)).

Solution The invention further relates to a medical solution including at least one sulphite compound to be included for the ability to reduce the concentration of decomposition products formed from a carbohydrate by stabilisation or scavenging already formed decomposition products, obtained during sterilisation and/or storage of solutions containing carbohydrate compounds, as defined above. Furthermore the "sulphite compound" could be used as an antioxidant or to scavenge toxic or allergenic compounds in vivo, such as in the body fluids.

The invention further relates to a medical solution such as a solution used for peritoneal dialysis either in a single or a multiple compartment container, comprising at least one sulphite compound to be included for the ability to reduce decomposition of a carbohydrate present in the solution exposed to sterilisation.

If a monosulphite compound such a bisulphite is used, it is preferably added to the carbohydrate and/or the electrolyte compartment in an amount to give a final solution with a concentration within the range of 0.01 10 mM, preferably 0.05 1 mM, most preferably about 0.05 0.5 mM. If a disulphite compound is used it is WO 01/89478 PCT/SE01/01125 preferably added to the carbohydrate compartment in an amount to give a final solution with a concentration within the range of 0.005 5 mM, preferably 0.025 mM, most preferably 0.025 0.25 mM.

If a disulphite compound is used it is preferably added in an amount to give a final solution which is half of the concentration used for the corresponding monosulphite compound.

The pH of the carbohydrate solution is not critical and could be in any range, suitable the pH is between pH 2.0-7.5, more preferably pH 2-5.5, even more preferably pH 3-4 and even more preferably about pH 3.2. The pH of the final solution is preferably between 5.0- 8.0 and more preferably 6.5 8.0 and most preferably 7.0 Preferably the medical solution is a sterile medical solution.

Additionally, the sulphite compound may be provided to the carbohydrate solution prior or after sterilisation.

Furthermore, the sterilisation is performed using any conventional sterilisation method as defined above under the term "sterilisation". Preferably the sterilisation is performed by heat treatment within the range of 100-150° C, for 1- 130 minutes, more preferably at 1210 C for 20 minutes (Ph. Eur. (current)).

The solution may be any medical solution which comprises a carbohydrate with or without other components. Preferably the solution is a medical solution such as a solution used for peritoneal dialysis, preferably medical solution(s) for single or multiple compartment container(s) for peritoneal dialysis, more preferably two or three compartment containers, even more preferably a three compartment container.

Medical solutions used for peritoneal dialysis preferably contain glucose in an amount to give a glucose concentration in the range of 1.5 to preferably about 2.5 or 4 by weight in the final solution (based on the total final solution).

Additionally the medical solution is a solution used to scavenge toxic or allergenic compounds in vivo, preferably a medical solution used to scavenge toxic or allergenic compounds in body fluids.

SMethod The invention further relates to a method for stabilising or scavenging the Sdecomposition of carbohydrate components produced in a medical solution during sterilisation and/or storage, including providing a sulphite compound to the solution prior or after the sterilisation process in order to reduce decomposition of 00 the carbohydrate components in the medical solution.

Preferably the method is used for preparation of medical solution(s) used C for peritoneal dialysis.

Even more preferably the medical solution is/are used in a multiple compartment container for peritoneal dialysis, such as a three compartment container.

Preferably, the method is used for preparation of a multiple compartment container used for peritoneal dialysis, wherein the sulphite compound may be added either to the carbohydrate compartment or to the electrolyte compartment of a multiple compartment.

If a monosulphite compound is used it is preferably added to the carbohydrate solution in an amount to give a concentration in the final solution within the range of 0.01 10 mM, preferably 0.05 1 mM, most preferably 0.05 mM.

If a disulphite compound is used it is preferably added to the carbohydrate solution in an amount to give a concentration in the final solution within the range of 0.005 5 mM, preferably 0.025 0.5 mM, most preferably 0.025 0.25 mM.

When a disulphite compound is used it is also preferably added in an amount to give a concentration in the final solution which is half of the concentration used for a corresponding monosulphite compound.

The pH of the carbohydrate compartment is preferably between pH more preferably pH 2-5.5, even more preferably pH 3-4 and most preferably about pH 3.2.

More preferably the method is used for the preparation of a sterile multiple compartment.

Additionally, the sulphite compound is provided to the carbohydrate solution prior or after sterilisation.

WO 01/89478 PCT/SE01/01125 12 Furthermore, the sterilisation is performed using any conventional sterilisation method as defined above under the term "sterilisation". Preferably the sterilisation is performed by heat treatment within the range of 100-1500 C, for 1- 130 minutes, more preferably at 121° C for 20 minutes (Ph. Eur. (current)).

The method according to the invention is intended to be used for medical solutions, in which the medical solution needs to be sterile, by a method as defined under the term "sterilisation", and preferably the method will be used for the preparation of medical solutions used for peritoneal dialysis or the like. By way of adding a sulphite compound in small amounts to the solution either prior or after sterilisation, decomposition of the carbohydrate components into toxic compounds in the solution is prevented or the toxic compounds are scavenged. Preferably, medical solutions to be used for peritoneal dialysis, preferably contain glucose in an amount to give a glucose concentration range between 1.5 to preferably about 2.5 or 4 by weight in the final solution.

Furthermore the sulphite compound could be used as an antioxidant or to scavenge toxic or allergenic compounds in vivo, such as in the body fluids.

Additionally the invention provides the use of a carbohydrate containing solution for the preparation of a multiple compartment container, preferably a three compartment container, suitable for peritoneal dialysis.

Specifically the invention provides the use of a carbohydrate containing solution for the preparation of a multiple compartment container for the treatment of an animal in need thereof.

MATERIALS AND METHODS Determination of glucose degradation products: Chemicals: Acetonitrile (Lab Scan, Ireland) and methanol (Lab Scan, Ireland) were of HPLC grade. 2,3- diaminonaphtalene was supplied by ICN, USA.

3-deoxyglucosone 56 (weight/weight) was synthesised by T. Henle Technische Univeritit Dresden. Sodiumphosphate p.a. and Glyoxal 30 (weight/volume) supplied by Merck (Germany), methylglyoxal 40 (weight/volume), 2,4-di- WO 01/89478 PCT/SE01/01125 13 nitrophenylhydrazine (2,4-DNPH) and 1,2-phenylenediamine were supplied by Sigma Chemical (USA). Acetaldehyde p.a. was supplied by Fluka (Germany).

Equipment: Two HPLC systems were used for the determination of glucose degradation products (GDP). One HPLC consisted of an Hewlett Packard liquid chromatograph serie 1050 equipped with an UV-detector and an autosampler. The second HPLC system consisted of an Hewlett Packard liquid chromatograph serie 1100 equipped with an autosampler and Waters Refractive Index detector model 410. Hewlett Packard Chem Station software rev. A.06.03, NT 4.0 was used for the data handling.

Determination of 3-deoxyglucosone 3-DG was determined using 2,3-diaminonaphtalene as derivative reagent. The samples were diluted 50 times to a total volume of 1 ml prior to analysis. The standards were prepared in the range 1-6 Standards and samples were prepared by adding 100 .10.1% (2,3diaminonaphtalene to 1 ml sample and incubated for 16 hours in room temperature in dark. The analytical column was a Water Symmetry C18 column (5 gm, 25 cm x4, 6 mm). The elution of the substance was performed at constant flow rate of ml/min by using a gradient of acetonitrile/water. The percentage of acetonitrile/water (volume/volume) was initially 25/75, and 12 minutes later 25/75, at 15 minutes 60/40 and at the gradient stop 30 minutes 60/40. The wavelength was set at 268 nm and the injected volume was 20 1l. The limit of quantification was 1 pM.

Determination of acetaldehyde and formaldehyde: The samples for the determination of acetaldehyde were diluted 20 times to a final volume of 4 ml, prior to analysis. Acetaldehyde was prepared as hydrazone derivatives using 2,4-DNPH as derivative reagent. The standards were prepared in range 1.1 11.4 M acetaldehyde, and 1.7-16.7 pM formaldehyde. Standards and samples were prepared by adding 2 ml 0.08 (weight/volume) 2,4-DNPH to 4 ml of each sample. The sample were concentrated on a solid phase extraction C18 column (Bond Elut LRC 200 mg/3 ml) and after rinsing with water, eluted with 1.6 ml acetonitrile. The analytical column was a Supelco C18 column (5 pm, 15 cm x 4,6 mm). The elution of the substances was performed at constant flow of 1.7 ml/min by using a linear WO 01/89478 PCT/SE01/01125 14 gradient of acetonitrile/water. The percentage acetonitrile/water (volume/volume) was initially 35/65 and at the gradient stop 12 minutes later 80/20. The wavelenght was set at 240 nm and the injected volume was 20 gl. The limit of quantification was for acetaldehyde. 1.1 gM. And for formaldehyde 1.7 pM.

Determination of glyoxal and methylglyoxal: Glyoxal and methylglyoxal were determined as quinoxalines using 1,2-phenylenediamine. The standards were prepared in the range 3.5-51.7 RM methylglyoxal. Standards and samples were prepared by adding 0.6 ml 0.4 (volume/volume) 1,2-phenylenediamine to 1 ml of each sample. The analytical column was Supelco C18 column (5 gm, 25 cm x 4,6 mm). The elution of the substances was performed at constant flow of 1.0 ml min using a mobile phase of initial 25 (volume/volume) acetonitrile and 75 (weight/volume) 0.05 M sodiumphosphate. At the gradient at 6 minutes the mobile phase was 30 acetonitrile and 70 millipore water and at gradient stop 9 minutes the percentages were 25/75. The wavelength was set at 312 nm and the injected volume was 20 pl. The limit of quantification for glyoxal was 3.5 |JM and for methylglyoxal 2.8 gM.

In vitro assay for cytotoxicity Medical solutions used for peritoneal dialysis were mixed with one part cell growth medium and 10% (volume/volume) fetal calf serum was added (Wieslander et al., 1991, Kidney Int. 40:77-79). Basal cytotoxicity of medical solution used for peritoneal dialysis were determined on mouse fibroblasts cells L-929 (CCL-1; ATTC, Rockville, MD, USA) as described earlier (Wieslander et al. 1993, Advances in Peritoneal Dialys, 9:31-35) and expressed as inhibition of cellgrowth

(ICG).

EXAMPLE 1.

Three compartment container with sulphite in the glucose compartment WO 01/89478 PCT/SE01/01125 A multiple compartment container as shown in FIG. 1, containing following medical solutions in the three compartments 44, 45 and 9.

Compartment 44 containing 100 ml of the composition: glucose calcium 20 mM magnesium 5 mM sodium 132 mM bisulphite 1 mM pH3.2 Compartment 45 contains 100 ml of the composition: glucose calcium 33 mM magnesium 8 mM sodium 132 mM bisulphite 1 mM pH 3.2 Compartment 9 contains 1900 ml with the composition: bicarbonate 40 mM sodium 132 mM pH 6.7 By mixing the contents of compartment 44 and compartment 9, a final solution suited for peritoneal dialysis is obtained with the following concentrations: glucose calcium 1.0 mM bicarbonate 38 mM sodium 132 mM WO 01/89478 PCT/SE01/01125 16 magnesium 0.25 mM bisulphite 0.05 mM By mixing the contents of compartment 45 and compartment 9, a final solution suited for peritoneal dialysis is obtained with the following concentrations: glucose calcium 1.65 mM bicarbonate 38 mM sodium 132 mM magnesium 0.4 mM bisulphite 0.05 mM By mixing the contents of both compartments 44 and 45 with the contents of compartment 9, a final solution suited for peritoneal dialysis is obtained with the following concentrations: glucose calcium 2.5 mM bicarbonate 36 mM sodium 132 mM magnesium 0.6 mM bisulphite 0.1 mM EXAMPLE 2.

Three compartment container with sulphite in electrolyte compartment A multiple compartment container as shown in FIG. 1, containing following medical solutions in the three compartments 44, 45 and 9.

WO 01/89478 PCT/SE01/01125 Compartnent 44 contains 100 ml of the composition: glucose calcium 20 mM magnesium 5 mM sodium 132 mM pH 3.2 Compartment 45 contains 100 ml of the composition: glucose calcium 33 mM magnesium 8 mM sodium 132 mM pH 3.2 Compartment 9 contains bicarbonate sodium bisulphite pH6.7 1900 ml with the composition: 40 mM 132 mM 0.1 mM By mixing the contents of compartment 44 and compartment 9, a final solution suited for peritoneal dialysis is obtained with the following concentration: glucose calcium 1.0 mM bicarbonate 38 mM sodium 132 mMI magnesium 0.25 mM bisulphite 0.095 mM WO 01/89478 PCT/SE01/01125 By mixing the contents of compartment 45 and compartment 9, a final solution suite for peritoneal dialysis is obtained with the following concentration: glucose calcium 1.65 mM bicarbonate 38 mM sodium 132 mM magnesium 0.4 mM bisulphite 0.095 mM By mixing the contents of both compartments 44 and 45 with the contents of compartment 9, a final solution suited for peritoneal dialysis is obtained with the following concentration: glucose calcium 2.5 mM bicarbonate 36 mM sodium 132 mMl magnesium bisulphite 0.6 mM 0.090 mM EXAMPLE 3 Two compartment container with sulphite in glucose compartment or three compartment container with both glucose compartments mixed.

The solutions used in example 3 and 4 were as described in example 1 and 2 except for that 50 glucose was used in all glucose compartments giving slightly different volumes for the compartments.

Two different sets of solutions were prepared, one electrolyte compartment with the volume 1.875 L and one glucose containing compartment with different amounts of bisulphite added, this volume was 125 ml. The solutions were sterilised at 1210 C for 1 hour and mixed post sterilisation. The concentrations post mixing of WO 01/89478 PCT/SE01/01125 19 electrolytes were 132 mM Na 1.35 mM Ca 0.25 mM Mg 2 95.2 mM Cl and mM lactate.

The concentration of glucose were 4 and the concentration of sulphite were 0. 0.01, 0.05, 0.1, 0.2 and 0.5 mM in the final solution.

The sterilised solutions were analysed in the In vitro assay ofcytotoxicity mentioned under Materials Methods. The results from the assay showed that an increase of the sodium bisulphite resulted in a decrease of the ICG value, which means that there is a decrease in the content of the toxic decomposition products after addition of a sodium bisulphite.

EXAMPLE 4 Two compartment container with sulphite in electrolyte compartment or three compartment container with both glucose compartments mixed As described in example 3 except for that sulphite was added to the electrolyte compartment. The concentrations of electrolytes, glucose and sulphite in the final solution were as in example 3. The sterilised solutions were analysed as in example 3. The results from the assay showed that an increase of the sodium bisulphite resulted in a decrease of the ICG value, which means that there is a decrease in the content of the toxic decomposition products.

EXAMPLE Analysis of a solution with or without sulphite for the presence of formaldehyde acetaldehyde and ICG.

Three solutions containing 132 mM Na 1.35 mM Ca 2 0.25 mM Mg 2 95.2 mM Cl, 40 mM lactate and 1.5 glucose was heat sterilised at 1210 C for 1 hour.

The sterilised solutions were mixed with sodium bisulphite to three different concentrations of sodium bisulphite 0.5, 1 mM).

Three solutions containing 132 mM Na 1.35 mM Ca 2 0.25 mM Mg 2 95.2 mM Cl-, 40 mM lactate and 1.5 glucose were mixed with sodium bisulfite to different concentration of sodium bisulphite 0.5, 1 mM) and was heat sterilised at 121 C for 1 hour.

SThe sterilised six solutions were analysed in the In vitro assay of N cytotoxicity and determination of acetaldehyde and formaldehyde mentioned 0 5 under Materials Methods. The results from the In vitro assay of cytotoxicity showed that an increase of the sodium bisulphite resulted in a decrease of the oo ICG value, which means that there is a decrease in the content of the toxic \decomposition products. The results from the assay determination of acetaldehyde and formaldehyde" showed decreased levels of both acetaldehyde C 10 and formaldehyde in the solutions containing sodium bisulphite, even down to levels under detection limit.

Comprises/comprising and grammatical variations thereof when used in this specification are to be taken to specify the presence of stated features, integers, steps or components or groups thereof, but do not preclude the presence or addition of one or more other features, integers, steps, components or groups thereof.

Claims (16)

1. Multiple compartment container for sterile medical solutions, said solutions 1 being present in two or more compartments, said container having breakable portions between two or more compartments, and at least one carbohydrate 00oo 5 compound is present in one or more of said compartments, and at least one sulphite compound is present in one or more of said compartments, wherein O when the breakable portions are broken the said solutions are mixed, (N wherein said sulphite compound is selected from the group consisting of HS0 3 52052 or SO 3 2 having a positive counter ion or mixtures thereof, and wherein if said sulphite compound is a monosulphite compound, the total amount of monosulphite compound in said container being equal to 0.05 1 mmol, per liter of the total amount of solution in said container, and if said sulphite compound is a disulphite compound, the total amount of disulphite compound in said container being equal to 0.025 0.5 mmol, per liter of the total amount of solution in said container.

2. Multiple compartment container according to claim 1, wherein said sulphite compound is NaHSO 3 Na 2 S 2 0 5 or Na 2 SO 3 or a mixture thereof.

3. Multiple compartment container according to claims 1 or 2, wherein if said sulphite compound is a monosulphite compound, the total amount of solution in said container is 0.05 0.5 mmol per liter.

4. Multiple compartment container according to any one of the above claims, wherein if said sulphite compound is a disulphite compound, the total amount to solution in said container is 0.025 0.25 mmol per liter. Multiple compartment container according to any one of the above claims, wherein the pH of the mixed solution in said container is between 5.0 8.0, more preferably between 6.0 8.0, most preferably between 7.0

6. Multiple compartment container according to any one of the above claims, c wherein said carbohydrate is glucose and the total amount of glucose in said container is about 4% by weight of the total amount of solution in said container. (Ni

7. Multiple compartment container according to any one of the above claims, 0 5 wherein said carbohydrate compound is not present in all compartments, preferably in only one of two compartments in a two compartment bag or in only Stwo of three compartments in a three compartment bag.

8. Multiple compartment container according to claim 7, wherein the pH of said compartments containing said carbohydrate compound is between 2.0 and 7,5, preferably between 2.0 and 5,5, more preferably between 3 and 4 and most preferably about 3,2.

9. Multiple compartment container according to claim 7 or 8, wherein said sulphite is a monosulphite compound and wherein said concentration monosulphite after mixing of said different solutions give rise to a final solution with a concentration of monosulphite compound of 0.05 1mM, preferably 0.025 0.25 mM. Multiple compartment container according to claim 7 or 8, wherein said sulphite is a disulphite compound and wherein said concentration disulphite after mixing of said different solutions give rise to a final solution with a concentration of disulphite compound of 0.025 0.5 mM, preferably 0.025 0.25 mM.

11. Multiple compartment container according to any one of claims 7 to wherein said carbohydrate compound is glucose and said container is a three compartment bag and wherein the concentration of glucose after mixing of said different solutions give rise to a final solution in the. range of 1.5 to preferably about 1.5, 2.5 or 4% by weight of glucose based of the total amount of final solution.

12. Multiple compartment container according to any one of the above claims, wherein said container is sterilised using heat treatment. 0 13. Multiple compartment container according to any one of the above claims, N wherein said multiple compartment container contains medical solutions for Spreparing a final solution for peritoneal dialyses by mixing two or more of said N medical solutions. (N O 5 14. A sterile medical solution including a carbohydrate compound, at least one sulphite compound selected from the group consisting of HSO3, S 2 0 5 2- or SO32" having a positive counter ion or mixtures thereof, preferably NaHSO 3 Na 2 S 2 0s or Na 2 SO 3 or a mixture thereof, wherein said sulphite compound is a monosulphite Scompound, present in a concentration of between 0.05-1 mM or a disulphite compound, present in a concentration of between 0.025-0.5 mM. A sterile medical solution according to claim 14, wherein said sulphite compound is a monosulphite compound, present in a concentration of 0.05 mM.

16. A sterile medical solution according to claim 14 or 15, wherein said solution is heat sterilised.

17. A sterile medical solution according to any one of claims 14-16, wherein said solution is a solution for peritoneal dialysis.

18. A sterile medical solution according to any one of claims 14-17, wherein said solution is used in a three compartment container.

19. A method of stabilising a carbohydrate containing sterile medical solution wherein in that at least one sulphite compound is added to said solution in a concentration of 0.05 1 mM, preferably 0.05 0.5 mM if said sulphite component is a monosulphite compound, and in a concentration of 0.025 mM, preferably 0.025 0.25 mM if said sulphite component is a disulphite compound. A method according to claim 19, wherein said sulphite compound is selected from the group consisting of HSO3, S 2 0 5 2 or SO 3 2 having a positive O counter ion or mixtures thereof, preferably NaHSO 3 Na 2 S 2 0 5 or Na 2 SO 3 or a c mixture thereof. c 21. A method according to claim 19 or 20, wherein said solution is heat sterilised. oo

22. Use of a carbohydrate containing sterile medical solution according to any one of claims 14-18 for the preparation of a multiple compartment container according to any of claims 1-13. C

23. A multiple compartment container substantially as herein described with reference to the accompanying drawing. DATED this 1 1 th day of July 2005 GAMBRO LUNDIA AB WATERMARK PATENT TRADE MARK ATTORNEYS 290 BURWOOD ROAD HAWTHORN VICTORIA 3122 AUSTRALIA P21916AU00