LU83270A1 - STABLE CONCENTRATED SOLUTIONS OF CISPLASTINE IN A SOLVENT - Google Patents

Description

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The present invention relates to stable and concentrated solutions of cisplastin in polyethylene glycol, aqueous methoxy polyethylene glycol or mixtures thereof, containing a source of chloride ion.

Platinum compounds are a unique group of compounds in the v5 group of an antineoplastic genus. Rosenberg et al. in 1965 [Rosenberg, B. et al. Nature (London), 205, p. 698-699 (1965) 1 first noticed that they have an antibiotic effect, then that they constitute powerful antitumor agents In animals [Rosenberg, B. et al., Nature (London), 222, p. 385-386 (1969)] From a structural point of view, these are complexes formed by a cen-10 tral atom of platinum, surrounded by various arrangements of chlorine atoms or ammonia groups in either cis or trans position relative to plan.

Two of the most commonly studied platinum compounds are represented by the following diagrams: C1 15 Cl_NH3 Cl NH3 ΖΞ7 ΓΎ1

Cl nh3 Cl Rh3

Cl 20

Cis-platinum (II) Cis-platinum (IV)

Diaminedichloro-platinum (II) Diaminetetrachloro-platinum (IV)

As can be seen, the cis-diaminedichloro-platinum compound (II) has all of these chloro and amino groups in one plane. This compound, now known as cisplatin according to the terminology of the "United States Adopted Name" (USAN), was synthesized according to the following reaction: NH4 Cl

K2 [PtCM + 2NH, --- ^ cis- [Pt (NH3) CI,] + 2KCI

[See Kauffmart, G.B. et al., In "Inorganic Synthesis, J. Kleinberg (Ed.), P. 239-245, McGraw-Hill Book Co., Inc., New York, 1963].

30 v · 2 "Y.G. Breusova-Baidala et al. ” in "Akademia Nauk", u.R.s.s., No. 6, pages 1239-1242 (June 1974) describe the slow isomerization in aqueous medium of cis-diaminedichloro-platinum (il) in trans form.

J.W. Reishus and D.S. Martin "in the" Journal of the American Chemical 5 Society ", 83, p. 2457-1462 (1961) indicate the acid hydrolysis of cisplatin at 25 ° C and 35 ° C. These studies are carried out in aqueous solutions at concentrations of 1.5 x 10 ~ 3M, 2.5 x 10 ~ 3M and 5.0 χ 10 “3M, which corresponds to 0.45 0.75 and 1.5 mg / ml, respectively. The authors indicate that there is a certain ambiguity in the localization of the origin (that is to say the "zero point"), λ 10 for the hydrolysis curves, because it takes 10 to 30 minutes for the sample to completely dissolve, even at these low concentrations.

M. Rozencweîg et al. ” in "Annals of Internai Medicine", 86, p.803-812 (1977) review the results of various preclinical and clinical studies involving the use of cisplatin in experimental tumors in animals, as well as in various types of human tumors.

It should be noted that the study drug, available to qualified researchers through the "Investigational Drug Branch of the Cancer Therapy Evaluation Program of the National Cancer Institute", is provided. as white lyophilized powder in vials containing 10 mg of cisplatin, 90 mg of sodium chloride, 100 mg of mannitol (U.S.P.) and hydrochloric acid to adjust the pH. When the product is reconstituted with 10 ml of sterile water for injection (U.S.P.), each ml of the resulting solution should contain 1 mg of cisplatin, 10 mg of mannitol and 9 mg of NaCl.

25 R.W. Talley et al. " in "Cancer Chemotherapy Reports", 57, p.465-471 (1973), describe the results obtained in Phase I of their clinical study concerning the use of cisplatin in the treatment of 65 human patients with a variety of neoplasms. As in the previous publication, the drug is supplied to them by the "National Cancer Institute" in 30 vials containing 10 mg of cisplatin, 90 mg of sodium chloride and v 100 mg of mannitol, so as to be able to reconstitute the product with 10 ml of sterile water.

A.H. Rossof et al. " in "Cancer", 30, p. 1451-1456 (1972) describe the results obtained by using cisplatin to treat 31 human patients with a variety of tumors. They note that the drug supplied by the "National Cancer Institute" is manufactured by "Ben Venue Laboratories, inc." and contains, per vial, 10 mg of cisplatin, 10 mg (sic) of mannitol and 9 mg (sic) of NaCl, and that the yellowish-white powder dissolves easily in 8 to 10 ml of sterile water.

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Certain information concerning the chemistry and pharmaceutical formulation of cisplatin is provided on pages 1 to 5 and 31 to 32 of the publication entitled: "CLINICAL BROCHURE, CIS-PLATINUM (II) DIAMINE-5 DICHLORURE (NSC-119875) 11, H. Haldelsman et al., "Investigational Drug

Branch, Cancer Chemotherapy Evaluation Program, Division of Cancer Treatment, National Cancer Institute (Revised August 1974). s On pages 31 and 32 thereof concerning the formulation of cisplatin supplied free of charge by the N.C.I. for clinicians to carry out their clinical evaluation in cancer chemotherapy, the following indications are noted: PHARMACEUTICAL DATA SHEET.

* NSC-119875 - Cis-diaminedichloroplatin (II).

15 Dose formulation 10 mg / vial: The content of each 20ml lead glass vial looks like a white freeze-dried cake Each vial contains 10 mg of NSC-119875, 90 mg of sodium chloride, 100 mg of mannitol and 20 hydrochloric acid to adjust the ph.

Preparation of the solution 10 mg / vial: When the product is reconstituted with 10 mg of sterile water for injection, USP, each ml of the resulting solution contains 1 mg of NSC-119875, 10 mg of mannitol and 9 mg of chloride sodium with a pH between 3.5 and 4.5.

Storage: Unopened dry vials should be stored at refrigeration temperatures (4 to 8 ° C).

Stability: The intact vials have a temporary stability of one year when stored at a refrigeration temperature (4 to 8 ° C).

Stability recommendations can be adjusted pending the completion of a two-year shelf life study.

The reconstitution carried out as indicated provides a pale yellow solution which is stable for a maximum of 1 hour at room temperature (22 ° C) when exposed to normal ambient lighting and a maximum of 8 hours at room temperature. room temperature (22 ° C) when protected from light. The reconstituted solutions can s 4

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refrigeration (4 to 8 ° C).

Caution: The lyophilized dose formulations contain no preservatives and it is therefore desirable to reject the solutions 8 hours after having reconstituted them.

British published patent application No. 2021946A describes stable aqueous solutions of cisplatin having a concentration of cisplatin between about 0.1 and 1.0 mg / ml and a pH between 2.0 and 3.0.

The solutions may also contain a non-toxic, pharmaceutically acceptable source of mineral chloride ions, such as sodium chloride, and an excipient, such as mannitol.

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The present invention relates to concentrated stable solutions of cisplatin having concentrations of between about 2.5 and 25 mg / ml. More particularly, the present invention relates to stable concentrated solutions of cisplatin in a solvent medium comprising approximately 30 to 95% of polyethylene glycol, having an average molecular weight of between approximately 150 and 9000, or else a methoxy polyethylene glycol having a weight average molecular weight between about 300 and 6000, or a mixture thereof, and about 5 to 70% water, these solutions also containing at least one pharmaceutically acceptable and non-toxic source of chloride ion , in an amount which is at least approximately equivalent to the amount of cisplatin present in the solution, and these solutions having a cisplatin concentration of between approximately 2.5 and 25 mg / ml.

Stable aqueous solutions of cisplatin have been described in British Patent Publication No. 2021946A. Although stable solutions containing concentrations of cisplatin up to approximately 1 mg / ml can be obtained in such an aqueous medium at room temperature, crystallization of cisplatin may occur when cold, for concentrations of cisplatin significantly above approximately 0.5 mg / ml.

It is not easy to redissolve this crystallized cisplatin by shaking the system at room temperature, although a solution can be obtained again by heating to about 37 ° C. Since transport and storage temperatures after sale cannot be controlled, and crystallization of cisplatin in the vials would cause undesirable problems for the physician using it, the maximum practical concentration of cisplatin in such aqueous media is approximately 0.5 mg / ml.

The cisplatin solutions according to the invention can contain up to about 25 mg of cisplatin per ml, although the preferred maximum is about 15 mg / ml. The solutions according to the invention containing 15 mg of cisplatin per ml were maintained at a temperature of 4 ° C for 12 months without any crystallization of cisplatin being observed.

Such solutions were also frozen at -60 ° C, then thawed at room temperature without any precipitation of cisplatin observed.

Thus, practical solutions prepared according to the invention can offer cisplatin concentrations at least 30 times higher than those which can be obtained from practical aqueous solutions prepared according to the prior art.

It will be appreciated that the concentrated solutions of cisplatin provided in accordance with the present invention require lower transportation and storage costs per unit dose than those involved for known aqueous solutions. Although the known freeze-dried solid form requires lower transportation and storage costs, this saving is more than offset by the time and expense involved in freeze-drying.

According to a preferred aspect of the present invention, the solvent medium comprises approximately 80 to 95% (and more preferably approximately 85 to 90%) of polyethylene glycol having an average molecular weight of between approximately 250 and 1600 (and more particularly between approximately 250 and 650) and about 25 5 to 20% (and more preferably, about 10 to 15%) of water.

According to an even more advantageous aspect of the invention, the solvent medium comprises approximately 90% of polyethylene glycol, having an average molecular weight between 350 and 450, and approximately 10% of water.

It is preferable that the solution contains about 5 to 20 mg of cisplatin per ml, and more preferably still about 10 to 15 mg / ml. The non-toxic, pharmaceutically acceptable source of chloride ion is preferably present at a concentration of at least about 2 equivalents per equivalent of cisplatin in the solution. Concentrations as high as 50 or more equivalents of chloride ion per equivalent of cisplatin may be used, depending on the concentration of cisplatin, the percentage of water present and the particular source of chloride ion, but also concentrations high in ion. chloride are generally neither necessary nor desirable.

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It will be noted by those skilled in the art, that in the case of a high concentration of cisplatin and a low water content, it would not be possible to dissolve a sufficient amount of chloride ion source such as sodium chloride, to provide 50 equivalents of chloride ion per 5 equivalents of cisplatin. In addition, a saturated or almost saturated solution. ture of a mineral chloride would be undesirable since it may crystallize in the solution when it is cold.

In the situation described above, 50 equivalents of chloride ion per equivalent of cisplatin could be obtained using hydrochloric acid as a source of chloride ion, but this could result in a solution having much too high acidity. , i.e. a low pH.

Applicants have found that excessively acidic solutions are somewhat less stable than more moderately acidic solutions.

The pH range of the solutions is preferably between approximately 1.5 and 4.5. The Applicant prefers to use approximately 2 to 10 equivalents of chloride ion per equivalent of cisplatin, and more preferably approximately 3 to 7 equivalents of chloride ion per equivalent of cisplatin.

The chloride ion can be provided by the addition of hydrochloric acid, a pharmaceutically non-toxic metal halide such as sodium, potassium, calcium or magnesium chloride, or a salt. of addition of hydrochloric acid to a pharmaceutically acceptable and non-toxic tertiary amine, such as triethylamine, or by mixtures thereof. The preferred source of ion; chloride is provided by hydrochloric acid, sodium chloride or a mixture thereof.

The polyethylene glycols and methoxy polyethylene glycols have the following general formulas: H (0CH2CH2) n0H and CH3 (0CH2CH2) n0CH3 \ respectively, and are available commercially under the names Polyethylene glycols CARBQWA) ^ and methoxy polyethylene glycols CARBOWA) ^ ®. The Applicant prefers to use the SENTRY grades of polyethylene glycols CARB0WAX, which are produced so as to meet the specifications U.S.P., N.F. and F.C.C. regarding food and drug applications.

Typical molecular weight ranges for a variety of CARB0WAX polyethylene glycols and CARB0WAX methoxy polyethylene glycols are provided in Tables I and II, below.

7 5 TABLE 1

Polyethylene Typical range

Glycols, | q CARBOWAX of Molecular Weight 200 190 to 210 300 285 to 315 400 380 to 420 15 600 570 to 630 1000 950 to 1050 1540 1300 to 1600 4000 3000 to 3700 6000 7000 to 9000 20 ___

TABLE II

25: Methoxy

Polyethylene Typical range

Molecular pQÎjg glycols

CARBOWAX

30 350 335 to 365 550 525 to 575 750 715 to 785 2000 1850 to 2150 35 8 * It is known that polyethylene glycols form complexes with certain mineral salts. Thus, the reaction of polyethylene glycols with ammonium cobalt thiocyanate forming a blue complex forms the basis of a colorimetric method for determining the concentration of polyethylene glycols in various mixtures. The Applicant thinks that the unique stability of cisplatin in the solutions according to the invention may be due to a complex formed between cisplatin and polyethylene glycol, but this is theoretical and does not constitute an element of the invention.

The proof of the formation of a complex was not noted in the chroma-10 tographies in thin layers carried out by the Applicant.

Using the techniques described below, the aqueous cisplatin produces a spot for an Rf value of approximately 0.65. However, a solution of cisplatin in 90% polyethylene glycol 400-10% water (or 10% 0.5N HCl) provides a spot for an Rf of about 0.3 which spreads to an Rf of about 0.65. Dilution with large quantities of water seems to immediately break the complex, since dilution of the above solution of PEG-H20 (or HCl) with five volumes of water then only shows a spot corresponding to cisplatin for an Rf d 'about 0.65.

20 Thin layer chromatography (TLC)

Apparatus and reagents - (a) TLC plates: 60 silica gel plates from EM Laboratories, Catalog No. 5753, or equivalent.

(B) Eluent - acetone: Normal nitric acid (9/1). Prepared fresh daily.

(c) Developer: Dissolve 5.6 g of stannous chloride in 10 ml of concentrated hydrochloric acid. Add 90 ml of distilled water and 0.2g of potassium iodide. Shake well. Prepare a 30 fresh product every day.

(d) Laboratory oven set at 100 ° C.

Procedure - (a) The sample is diluted with 5 volumes of dimethylformamide (DMF) [from Burdick and Jackson, and distilled in glass].

35 Cb) A TLC plate is stained with 5 microliters of the sample and 5 microliters of a standard solution containing cisplatin (and transplatinum and platinum B, if appropriate), at a concentration approximately equal to that which is expected in the sample to be analyzed. A height of 10 cm is revealed in the GCM reservoir 40 which is pre-liberated with Véluent.

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The dried plate is sprayed with the freshly prepared developer solution and placed in an oven at 100 ° C for 10 minutes.

We observe the yellow / purple zones.

(c) Approximate Rf values: 5 · 0.65: cisplatin; . 0.76: transplatin,. 0.90: platinum B.

Titrations by HPLC of the solutions according to the invention tending to determine the cisplatin content can be carried out according to the procedure described in British patent No. 2021946A. The preferred mobile phase consists of a mixture of ethyl acetate / methanol / dimethylformamide / distilled water (25/26/5/5). The standard is preferably constituted by cisplatin dissolved in dimethylformamide at a concentration of 1 mg / ml. The samples to be analyzed are diluted with dimethylformamide to an approximate cisplatin concentration of 1 mg / ml.

Although their presence does not confer any particular advantage, the solutions according to the invention may optionally contain a conventional excipient which is acceptable from a physiological point of view such as mannitol.

From the stability studies carried out at present, it is predicted that the stability of the solutions according to the invention (as being a loss of 10¾ of power) is greater than two years at room temperature.

According to a preferred aspect of the invention, the solutions are sterile and devoid of pyrogenic substance; they are packaged in sterile containers and devoid of pyrogenic substance. Such solutions can then be diluted with, for example, sterile water for injection, U.S.P. grade, or sterile normal saline, U.S.P. grade, and administered intramuscularly or intravenously.

The means for sterilizing such solutions are well known in the art. The Applicant prefers to pass the solutions I through a 30 millipore 0.22 micron filter, sterile and devoid of pyrogenic substance, using aseptic techniques, under sterile nitrogen pressure. Mi llipore is a trade name of the Millipore Corporation for membrane filters. The sterile filtrate is collected in sterile containers devoid of pyrogenic substance, then finally introduced in desired quantity, in suitable sterile receptacles, devoid of pyrogenic substance, closed with sterile stoppers and devoid of pyrogenic substance (preferably coated with Teflon) and sealed with sterile aluminum capsules.

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To use them in the treatment of cancer, the concentrated solutions are diluted to the desired concentration (generally 1 mg of cisplatin per ml) with for example sterile water for injection, of USP quality, normal sterile saline solution, of USP, or sterile glucose solution, used by intravenous or intramuscular injection, or by intravenous infusion, as hitherto known for administration of cisplatin preparations.

Doses commonly used, having an acceptable to moderately acceptable mild toxicity, are between 60 and 100 mg / m2 intravenously, 10 administered in a single dose, or distributed over 3 to 5 days, this dose having to be re-administered at intervals of 4 weeks. A dose of 60 mg / m2 is roughly equivalent to a dose of 1.5 mg / kg which, in turn, is roughly equivalent to a dose of 105 mg for a patient weighing 70 kg.

Concurrent therapy with other chemotherapeutic agents is frequently prescribed in order to achieve the best results.

As used herein in the claims, references to "equivalents" of chloride ion by "equivalent" of cisplatin mean molar equivalents. Thus, for example, when using the preferred range of from about 3 to 7 equivalents of chloride ion per equivalent of cisplatin, about 3 to 7 moles of NaCl would be used per mole of cisplatin but about 1.5 to about 3 , 5 moles of CaCl2 per mole of cisplatin, etc.

Platinum B is an arbitrary designation used here for the acid reaction product of cisplatin which is half the known red complex of Magnus 25 and to which we have tried to attribute the following structure:

Cl ^ Pt ^ _ h3N ^^ 'V ^ cl _

This invention is illustrated but cannot be limited by the following Examples.

EXAMPLE 1. · Stable concentrated solution of cisplatin (10 mg / ml) in 90% polyethylene glycol 400 - 10% normal HCl - 35 A slurry is made with 500 mg of cisplatin in a solution of 5 ml of normal hydrochloric acid and 45 ml of polyethylene glycol 400.

After 2 days of stirring at room temperature (the container being protected from light with an aluminum sheet), a light yellow solution is obtained.

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The aliquots of this solution are introduced into 17 ml amber flasks, hermetically closed with caps coated with teflon, sealed with aluminum capsules and subjected to storage stability tests at various temperatures. After storage for two weeks at 56 ° C, thin layer chromatography (TLC) indicates the presence of a trace of transplatin and approximately 1% platinum B. After storage for two weeks at 56 ° C, TLC indicates the presence of less than 1% of transplatin and approximately 3¾ of platinum B.

The samples stored for two weeks at 56 ° C. are diluted with, respectively, 4, 9 and 19 volumes of water, and provide transparent solutions containing, respectively, 2, 1 and 0.5 mg / ml of cisplatin.

A slight cloud appears in each of the diluted samples after one. rest about 18 hours at room temperature.

EXAMPLE 2 15 · Stable concentrated solution of cisplatin (10 mg / ml) in 90% polyethylene glycol 400 - 10% 0.5N HCl - 5 ml of purified water of U.S.P quality and 5.0 ml of normal HCl are mixed and added with 90.0 ml of polyethylene glycol 400. To 50 ml of the above solution are added 500 mg of cisplatin and the mixture is.

20 protected from light with aluminum foil and stirred at room temperature for 24 hours to produce a clear solution.

The TLC of a freshly prepared solution shows only one area of cisplatin with a possible trace of transplatin.

Two 2 ml aliquots of the solution are introduced into 17 ml amber vials, closed with caps coated with teflon and sealed with aluminum capsules, then subjected to storage stability tests at various temperatures.

A sample vial is lyophilized in a dry ice-acetone bath for 1 hour, then allowed to warm to room temperature.

• 30 A clear solution is obtained, and no precipitate is observed. After storage for 3 months at 37 ° C and 45 ° C, the TLC indicates the presence of 1 to Zi of platinum B and a possible trace of transplatin.

After 1 month of storage at 56 ° C, the TLC indicates the presence of more than 5% but less than 10¾ of platinum B and a trace of transplatin.

The samples stored at 37 and 45 ° C for 3 months, and at 56 ° C for 1 month, are diluted with four volumes of purified water of U.S.P quality. to give clear solutions containing 2 mg / ml of cisplatin.

The diluted solutions remain clear after standing for 24 hours at room temperature.

t EXAMPLE 3 12 • Stable concentrated solution of cisplatin (10 mg / ml) + CaCl2 (20 mg / ml) in 90% polyethylene glycol 400 - 10% HCl 0 »5N -

2 g of anhydrous reactive grade calcium chloride are dissolved in a mixture of 5 ml of purified water of U.S.P grade. and 5 ml of normal HCl. 89 ml of polyethylene glycol 400 are added to bring the volume to 100 ml.

To 50 ml of this solution is added 550 mg of cisplatin, and the mixture is protected from light with aluminum foil; the system is stirred at room temperature for 24 hours to obtain a clear solution.

The TLC of a freshly prepared solution indicates only the presence of a zone of cisplatin with a possible trace of transplatin.

2 ml aliquots of the solution are placed in 17 ml amber flasks, closed with Teflon coated stoppers, sealed with aluminum capsules and subjected to storage stability tests at various temperatures.

A sample flask is placed in a dry ice-acetone bath for 0.5 hour and lyophilized in a transparent gel. This is allowed to warm to room temperature and a clear solution is collected. 20 After storage for 3 months at 37 ° C, the TLC indicates the presence of 2% platinum B and a possible trace of transplatin.

After storage for 3 months at 45 ° C., the TLC indicates the presence of approximately 5% of platinum B and a possible trace of transplatin.

After storage for 1 month at 56 ° C., the TLC indicates the presence of 8 to 10% of platinum B and a trace of transplatin.

The samples stored at 37 ° C and 45 ° C for 3 months, and at 56 ° C for 1 month, are diluted with 4 volumes of purified water of U.S.P quality. to give clear solutions containing 2 mg / ml of cisplatin.

The diluted solutions remain clear after standing for 24 hours at room temperature.

EXAMPLE 4 • Stable concentrated solution of cisplatin (approximately 22 mg / ml) + CaCl2 (25 mg / ml) in 90% polyethylene glycol 400 - 10% 0.5N HCl - 35 3 ml of a 90% solution of Polyethylene glycol 400 and 10% 0.5N HCl are added with 45 mg of cisplatin, and the mixture is stirred for 1 hour at room temperature to give a clear solution.

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An additional dose of 30 mg of cisplatin is introduced (making a total of 25 mg / ml) and the mixture is stirred at approximately 45 ° C for 1 hour, then at room temperature for 16 hours, to produce a substantially complete solution. The small amount of insoluble matter is removed by filtration. The TLC of the filtrate indicates only a zone of cisplatin with a possible trace of transplatin. The remainder of the filtrate is introduced into a 17 ml amber flask, closed with a Teflon-coated stopper, sealed with an aluminum capsule and kept at 45 ° C for 3 months.

After aging for 3 months at 45 ° C, the TLC indicates that there is 1 to 2¾ of 10 platinum B and no transplatin in the solution.

A dilution of the aged solution with purified water of U.S.P quality. at a concentration of 2 mg / ml of cisplatin provides clear solutions, which remain so after standing at room temperature for 24 hours.

EXAMPLE 5 15 • Stable concentrated solution of cisplatin (12 mg / ml) + NaCl (10 mg / ml) in 90% polyethylene glycol 400 - 10% HCl 0 »5N -

100 mg of sodium chloride is dissolved in a solution of 5 ml of purified water of U.S.P quality. and 5 ml of normal hydrochloric acid. To this solution is added 90 ml of polyethylene glycol 400 and the mixture is stirred for 15 minutes. 50 ml of cisplatin are added to 50 ml of the solution obtained, and the mixture is stirred in the dark at room temperature for 3 days, to produce a clear solution.

. The TLC of the freshly prepared solution shows only one spot corresponding to cisplatin. A titration by high performance liquid chromatography (HPLC) of the freshly prepared solution indicates that it contains 12 mg of cisplatin per ml.

Aliquots are introduced into 17 ml flasks of amber, the latter being sealed as described in Example 3, and subjected to storage stability tests at various temperatures.

After storage at 37 ° C for 3 months, the TLC indicates that there is less than 1% of platinum B and no transplatin.

After storage at 56 ° C for 1 month, the TLC indicates that there is 1-2% platinum B and no transplatin.

35 Diluting aged samples with purified U.S.P. water up to a concentration of 2 mg / ml of cisplatin provides clear solutions which remain so after standing at room temperature for 24 hours.

HPLC titrations of samples stored for 3 months at 45 ° C, 6 months at 37 ° C and 8 months at room temperature indicate 40 power losses of 6.6%, 6.1% and 2.3% , respectively.

EXAMPLE 6 14 • Stable concentrated solution of cisplatin (11 »4 mg / ml) + NaCl (10 mg / ml) in 90% polyethylene glycol 400 - 10% water ™

A solution of 50 mg of NaCl in 5 ml of purified water of U.S.P. quality and 5 of 45 ml of polyethylene glycol 400, is added with 500 mg of cisplatin, and the mixture is stirred, in the dark, at room temperature, for 6 h to provide a clear solution.

The TLC of the freshly prepared solution reveals the presence only of the stain corresponding to cisplatin; HPLC titration indicates that this solution contains 11.4 mg of cisplatin per ml.

Aliquots are introduced into 17 ml amber vials and these are sealed as described in Example 3, then subjected to storage stability tests at various temperatures.

After storage at 37 ° C and 45 ° C for 3 months, the TLC indicates that there is 1% platinum B and no transplatin.

After storage at 56 ° C for 1 month, the TLC indicates that there is 2 to 3% of platinum B and no transplatin.

A dilution of the aged samples with purified water of U.S.P quality at a concentration of 2 mg per ml of cisplatin provides clear solutions which remain so after standing at room temperature for 24 hours.

HPLC titrations of samples stored 3 months at 45 ° C, 6 months at 37 ° C and 7 months at room temperature reveal power losses of 6.1%, 7.0 and 0.9%, respectively.

EXAMPLE 7 25 • Stable concentrated solution of cisplatin (10 mg / ml) in 90% polyethylene glycol 600 - 10% 0.5N HCL -

A solution of 2.5 ml of purified water of USP quality, 2.5 ml of normal HCl and 45 ml of polyethylene glycol 600 is added with 500 mg of cisplatin and the mixture is stirred in the dark at room temperature, for 5 hours, to provide a clear solution.

The TLC of the freshly prepared solution indicates only one spot corresponding to cisplatin.

Samples of the freshly prepared solution are diluted with 1, 2, 3, 4, 5 and 9 volumes of purified U.S.P. water, respectively; these dilute solutions show no crystallization, after standing for 16 hours at room temperature or at 4 ° C.

The aliquots of the freshly prepared solution are introduced into 17 ml amber flasks, which are sealed as described in Example 3 and then subjected to storage stability tests at various temperatures.

After storage at 37 ° C and 45 ° C for 3 months, the TLC indicates that there is 1% platinum B and no transplatin.

5 After storage at 56 ° C for 1 month, the TLC indicates that there is 3.4¾ of platinum B and no transplatin.

Diluting aged samples with purified U.S.P. water at a concentration of 2 mg / ml of cisplatin provides clear solutions, which remain so after standing at room temperature for 24 hours.

10 EXAMPLE 8 • Stable concentrated solution of cisplatin (10 mg / ml) + NaCl (10 mg / ml) in 90% polyethylene glycol 400 - 10% 0.2N HCL - *

250 mg of cisplatin is added to a solution of 0.5 g of NaCl in 4 ml of purified water of USP quality, 1 ml of normal hydrochloric acid and 45 ml of polyethylene glycol 400. dark at room temperature for 4 hours to provide a clear yellow solution. The TLC of the freshly prepared solution indicates only one spot corresponding to cisplatin. Dilutions of the freshly prepared solution with 1, 2, 3, 4, 5 and 9 volumes of purified water provide clear solutions, which remain so after standing at room temperature for 24 hours. Dilutions with 1, 2, 3, 4 and 5 volumes show no crystallizations when kept at 4 ° C for 24 hours.

Aliquots of the freshly prepared solution are introduced into 17 ml amber vials, which are sealed as described in Example 3, and subjected to storage stability tests at various temperatures.

After storage at 37 ° C for 3 months, the TLC indicates that there is 1¾ of platinum B and a possible trace of transplatin.

After storage at 45 ° C for 3 months, the TLC indicates that there is 5% platinum B and a possible trace of transplatin.

After storage at 56 ° C for 1 month, the TLC indicates that there is 2 to 3% of platinum B and no tran.splatine.

Diluting aged samples with purified U.S.P. water 35 at a concentration of 2 mg / ml of cisplatin, provides clear solutions which remain so after standing for 24 hours at room temperature.

EXAMPLE 9 16 • Stable concentrated solution of cisplatin (2.5 mg / ml), NaCl (9 mg / ml) and mannitol (12.5 mg / ml) in 31% (weight / volume) of acidified aqueous polyethylene glycol 6000 - 5 0.9 g of sodium chloride, 1.25 g of mannitol and 31.3 g of polyethylene glycol are dissolved in sufficient purified water of USP quality to - have 100 ml of a solution, which is acidified to a pH of 2.2 using normal hydrochloric acid (0.7 ml). 255 mg of cisplatin is added and the mixture is stirred in the dark for 3 days at room temperature to obtain a clear solution.

The TLC of the freshly prepared solution only shows the zone corresponding to cisplatin.

Aliquots of the freshly prepared solution are introduced into 17 ml amber vials, which are sealed as described in Example 3, and then subjected to storage stability tests at 45 ° C and 56 ° C. After 2 months of storage at 45 ° C and 56 ° C, the TLC indicates that there is less than 1% of platinum B and no transplatin.

Diluting aged samples with an equal volume of purified U.S.P. water provides clear solutions, which remain so after standing at room temperature for 24 hours.

EXAMPLE 10 • Stable concentrated solution of cisplatin (15.8 mg / ml) + NaCl (10 mg / ml) in 90% aqueous polyethylene glycol 400 - 25 90 ml of polyethylene glycol 400 are dissolved in a solution of 1.0 g of NaCl in 10 ml of USP grade purified water 900 ml of cisplatin are added to 60 ml of the solution obtained, and the mixture is stirred in the dark for 5 hours at room temperature to obtain a clear solution.

The TLC of the freshly prepared solution indicates only the presence of the zone corresponding to cisplatin; a titration by HPLC indicates that it contains 15.8 mg of cisplatin per ml.

Diluting the freshly prepared solution with 4 volumes of purified U.S.P. quality water provides a clear solution, which remain after standing at room temperature for 24 hours.

Aliquots of. the freshly prepared solution are introduced into 17 ml amber flasks, which are then sealed as described in Example 3, then subjected to storage stability tests at various temperatures.

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After 2 months of storage at 45 ° C, the TLC indicates that there is 1.5% platinum B and no transplatin.

After 1 month of storage at 56 ° C, the TLC indicates that there is 2% platinum B and no transplatin.

5 A sample aged at 56 ° C is diluted to a concentration of 2 mg per ml with sterile water for injection and the sample aged at 45 ° C is diluted to a concentration of 2.5 and 5.0 mg / ml of cisplatin with sterile water for injection. They each provide clear solutions, which remain so after standing at room temperature for 24 hours.

10 HPLC titrations of samples stored 3 months at 45 ° C, 6 months at 37 ° C and 8 months at room temperature indicate power losses of 7.6%, 7.6% and 0%, respectively.

EXAMPLE 11 jg · Stable concentrated solution of cisplatin (15 mg / m) + CaCl (25 mg / ml) in 90% aqueous polyethylene glycol 400 -

A solution of 2.5 g of CaCl2 in 10 ml of purified water of U.S.P. quality 90 ml of polyethylene glycol 400 are added and the resulting solution is stirred for 10 min. To 50 ml of the above solution, 750 mg of cisplatin is added and the mixture is stirred for 5 hours in the dark, at room temperature, to obtain a clear solution.

The TLC of the freshly prepared solution indicates only an area corresponding to cisplatin. Dilutions of the freshly prepared solution with 1, 2, 5 and 10 volumes, respectively, of purified water of U.S.P quality.

25 provide a clear solution.

Aliquots of freshly prepared solution are introduced into 17 ml amber vials, which are sealed as described in Example 3, then subjected to the storage stability tests at 45 ° C and 56 ° C. After storage for 2 months at 45 ° C, the TLC indicates that there is 1.5% platinum B 30 and no transplatin.

After storage for 1 month at 56 ° C, the TLC indicates that there is 2.5 to 5% platinum B and no transplatin.

A sample aged at 56 ° C is diluted to a concentration of 2 mg / ml and the sample aged at 45 ° C is diluted to concentrations of 2.5 and 5.0 mg / ml of cisplatin, with sterile water for injection.

They both form clear solutions, which remain after standing at room temperature for 24 hours.

♦ EXAMPLE 12 18 • Sterile stable concentrated solution of cisplatin in 90% polyethylene glycol 400 - 10% HCl 0 »5N (The label indicates 15 mg / ml of cisplatin activity) - 5 note: Cisplatin is a carcinogenic agent possible.

Protective clothing, gloves, mask, goggles and hat must be worn during the entire procedure.

All work areas and equipment must be thoroughly cleaned to avoid future contamination.

10

FORMULA

per ml per 10.0 ml

Cisplatin ........... 0.015 g 0.150 g 15

Sodium chloride ..... 0.015 g 0.150 g

0.5N hydrochloric acid 2) ........... 0.10 ml 1.0 ml

Polyethylene glycol 400 (SENTRY quality) q.s.p. 1.0 ml q.s.p. 10.0 ml 20 1> This weight of cisplatin provides a power of 1000 micrograms / mg. To determine the quantity of cisplatin required, the following formula is used: -: - 1 ° PQ χ .β? Β1.§6 -; --— = Grams of cisplatin required 25 power of cisplatin in mcg / mg ^ 2) 1 liter of 0.5N hydrochloric acid is prepared as follows: v 1. 957.25 ml of sterile water for injection, of USP quality, is introduced in an Erlenmeyer flask by H.

30 ..., 2. With rapid stirring, slowly and carefully introduced 2.75 ml of concentrated hydrochloric acid. Stirred further for 10 min. Close with a clean butyl rubber stopper.

IH

MANUFACTURING INSTRUCTIONS FOR PREPARING A LITER OF STERILE SOLUTION.

1 - 100 ml of 0.5N hydrochloric acid are placed in a clean Erlenmeyer flask calibrated at 1 £, containing a suitable stirrer such as a 6 cm magnetic stirrer bar, coated with Teflon.

5 2 - 15.0 g of sodium chloride are added, with moderate stirring.

Stirring is continued until complete dissolution.

3 - With rapid stirring, 750 ml of polyethylene glycol 400 (SENTRY quality) are introduced and stirring is continued for 5 min.

jq 4 · - The magnetic bar is removed and it is drained so that the excess fluid falls back into the falcon.

5— Carefully add 15.0 g of cisplatin activity.

b - Polyethylene glycol 400 (SENTRY quality) is added up to the mark l £ (A total of 880 ml of polyethylene glycol 400 is required).

7- The teflon stirrer is returned to the mixture and the system is closed with a clean butyl rubber stopper.

8— The container is wrapped in aluminum foil to protect it from light.

20 R; - Stir quickly for 24 to 48 hours at room temperature.

We must get a clear solution. If you don't get one. clear solution in 48 hours, the mixture can be heated to 37-40 ° C for 2 to 6 hours, in the absence of air and light, to facilitate the rapid dissolution of the remaining cisplatin. Cool to 23-27 ° C.

- Using an aseptic technique, the dark yellow solution is passed under a suitable sterile nitrogen pressure, through a suitable 0.22 micron Millipore filter, sterile and free of pyrogenic substance. The sterile filtrate is collected in a sterile Erlenmeyer flask devoid of pyrogenic substance. The flask is closed with a sterile stopper, devoid of pyrogenic substance, made of butyl rubber.

The solution should be stored in the dark.

It— The necessary quantity of sterile solution is introduced into sterile amber vials devoid of pyrogenic substance. These vials are closed with a sterile Teflon stopper and devoid of pyrogenic substance. These flasks are closed hermetically with al umi ni um s tér-i capsules.

20 12— Vials must contain the following warning label:

PRODUCT NOT TO BE USED DIRECTLY INTRAMUSCULARLY

OR INTRAVENOUS * * The PEG 400 solution can be diluted with 1L parts of sterile water 5 for injection of U.S.P quality. or sterile normal U.S.P. to provide a 1 mg / ml solution of cisplatin. If higher concentrations are required, proportionately less sterile water or saline may be used. The diluted solutions can be used intravenously and are stable at room temperature (22 to 26 ° C) for at least L8 hours. Do not put diluted solutions in the refrigerator • as crystals may appear.

13— Keep the vials in the dark.

EXAMPLE 13 • Stable concentrated solution of cisplatin (2.5 mg / ml), NaCl (9 mg / ml),

CaCl2 (15 mg / m) and mannîtol (10 mg / ml) in 31% acidified aqueous polyethylene glycol 400 - i 20 0.9 g of sodium chloride, 1.5 g of CaCl2 and 1.0 g of mannitol are dissolved in mixture of 31.25 ml of polyethylene glycol 400 and 40 ml of purified water of USP quality The solution is acidified to a pH of 2.2 using normal hydrochloric acid (0.4 ml), then 255 mg of cisplatin is added and the volume is brought to 100 ml with purified water of USP quality ; The mixture is stirred in the dark for 5 hours at room temperature to obtain a clear solution.

The TLC of the freshly prepared solution indicates only one zone corresponding to cisplatin.

EXAMPLE 14 The general procedure described in Example 5 is repeated, except that the sodium chloride is replaced by an equivalent amount of magnesium chloride; a stable concentrated solution of cisplatin is thus obtained.

EXAMPLE 15 The general procedure described in Example 5 is repeated except that the sodium chloride is replaced by an equivalent amount of triethylamine hydrochloride; οη thus obtains a concentrated solution λ <Ι · ^ Ι \ 1 λ i4 / \ / »η λιλ I 4 · η λλ EXAMPLE 16 c 21

The general procedure described in Example 5 is repeated except that the polyethylene glycol 400 is replaced by an equal volume of polyethylene glycols 200 and 300, respectively. Stable concentrated solutions of cisplatin are obtained.

EXAMPLE 17

The general procedure described in Example 9 is repeated except that the polyethylene glycol 6000 is replaced by an equal weight of polyethylene glycols 1000 and 4000 respectively. Concentrated very stable solutions of cisplatin are obtained.

Of course, the present invention is in no way limited to the examples * and modes of implementation mentioned above; it is susceptible of numerous variants accessible to those skilled in the art, depending on the applications envisaged and without departing from the spirit of the invention.

♦

Claims (8)

22 * 1. “Stable concentrated solution of cisplatin in a solvent medium, characterized in that it comprises approximately 30 to 95% of polyethylene glycol having an average molecular weight of between approximately 150 and 9000, or a methoxy polyethylene glycol having a average molecular weight of between about 300 and 6000, or a mixture thereof, and about 5 to 70% of water, and moreover also at least one source of chloride ion which is non-toxic and acceptable from the point of view pharmaceutical, in an amount which is at least about equivalent to the amount of cisplatin present in the solution, this solution having a cisplatin concentration of between about 2.5 and 25 mg / ml. 2. Stable concentrated solution of cisplatin in a solvent medium, characterized in that it comprises approximately 80 to 95% of polyethylene glycol having an average molecular weight of between approximately 250 and 1600, or a methoxy polyethylene glycol having an average molecular weight between about 300 and 800, or a mixture thereof, and about 5-20% water, as well as at least one non-toxic and pharmaceutically acceptable source of chloride ion, in an amount which is between about one equivalent and ten equivalents per · 20 equivalent of cisplatin to the solution, this solution containing approximately 5 to 20 mg of cisplatin per ml. 3. Cisplatipe solution according to claim 2, characterized in that the non-toxic and pharmaceutically acceptable source of chloride ion consists of hydrochloric acid, sodium chloride, calcium chloride, magnesium chloride, or triethylamine hydrochloride, or a mixture thereof. 4. A stable concentrated solution of cisplatin in a solvent medium, ♦ characterized in that it comprises approximately 85 to 90% of polyethylene glycol having an average molecular weight of between approximately 250 and 650, and approximately 10 to 15% of water, and also a source of chloride ion selected from the group consisting of hydrochloric acid, sodium chloride or mixtures thereof, in an amount which is between about 2 and 7 equivalents per equivalent of cisplatin in the solution, this solution having a concentration of cisplatin of between approximately 10 and 15 mlg / ml. 5. Cisplatin solution according to claim 4, characterized in that the solution is sterile and is in a hermetically sealed container, such as a vial. 23% 6. Sterile stable concentrated solution of cisplatin in a hermetically closed container such as a vial, characterized in that it contains approximately 10 to 15 mg of cisplatin per ml, and approximately 10 to 15 mg of NaCl per ml in a solvent medium. comprising about 90¾ of a polyethylene glycol having an average molecular weight of between about 350 and 450, and about 10¾ of water. 7. Sterile stable concentrated solution of cisplatin in a sealed container such as a vial, characterized in that it contains approximately 10 to 15 mg of cisplatin per ml, and approximately 10 to 15 mg of NaCl per ml in a solvent medium consisting of approximately 90¾ of a polyethylene glycol having an average molecular weight of between approximately 350 and 450, and approximately 10¾ of dilute hydrochloric acid, the concentration of which can range up to 0.5N. 8. Pharmacological application of the stable concentrated solutions according to any one of claims 1 to 7, in particular for cancer therapy.