FR2480605A1 - CONCENTRATED STABLE CISPLASTIN SOLVENT SOLUTIONS AND THEIR PHARMACOLOGICAL APPLICATION - Google Patents

Abstract

THE SUBJECT OF THE PRESENT INVENTION IS A STABLE CONCENTRATED SOLUTION OF CISPLATIN IN A SOLVENT MEDIUM. IT IS CHARACTERIZED IN THAT IT CONSISTS OF 30 TO 95 OF POLYETHYLENE GLYCOL OR METHOXY POLYETHYLENE GLYCOL, 5 to 70 OF WATER AND AT LEAST ONE SOURCE OF NON-TOXIC AND PHARMACEUTICALLY ACCEPTABLE CHLORIDE ION. THE INVENTION APPLIES TO A STERILE STABLE CONCENTRATED SOLUTION OF CISPLATIN CONTAINED IN A TIGHTLY CLOSED CONTAINER SUCH AS A FLASK.

Description

STABLE CONCENTRATE SOLUTIONS OF CISPLASTIN IN A SOLVENT

  AND THEIR PHARMACOLOGICAL APPLICATION.

  The present invention relates to stable and concentrated solutions of cisplastine in polyethylene glycol, aqueous methoxy polyethylene glycol

  or mixtures thereof, containing a source of chloride ion.

  Platinum compounds constitute a unique group of compounds in the group of an antineoplastic genus.Rosenberg et al. in 1965 [Rosenberg, B. et al. Nature (London), 205, p. 698-699 (1965)] first noted that they exhibit an antibiotic effect, and that they are potent 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-

  platinum, surrounded by various arrangements of chlorine atoms or

  ammonia groups in either cis or trans position relative to the plane.

  Two of the most commonly studied platinum compounds are represented by the following diagrams:

C NH3 CI NH3

CINH Pt t

NH3 CI H3 CI

  Cis-platinum (II) Cis-platinum (IV) Diaminedichloro-platinum (II) Diaminetetrachloroplastic (IV) As can be seen, the compound cis-diaminedichloro-platinum (II) has all these groups chloro and amino in a single plane. This compound, now known as cisplatin according to the United States Adopted Name (USAN) terminology, was synthesized according to the following reaction: NH, Cl K2 [PtCl4 + 2NH3 ac- [Pt (N1t3) Cl2] + 2KCI [See Kauffmari, GB et al., In Inorganic Synthesis, J. Kleinberg (Ed.),

  p. 239-245, McGraw-Hill Book Co., Inc., New York, 1963].

2 2480605

  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 (II) in trans form.

  J. W. Reishus and D. S. Martin, in the Journal of the American Chemical Society, 83, p. 2457-1462 (1961), indicate the acid hydrolysis of cisplatin

  at 25 ° C. and at 35 ° C. These studies are conducted in aqueous solutions at concentrations

  1.5 x 10-3M, 2.5 x 10-3M and 5.0 x 10-3M, corresponding to 0.45 0.75 and 1.5 mg / ml, respectively. The authors indicate that there is some ambiguity in the location of the origin (ie the "zero point"), for the hydrolysis curves, because it takes 10 to 30 minutes' so that

  the sample dissolves completely, even at these low concentrations.

  M. Rozencweig et al., In Annals of Internal Medicine, 86, 803-812

  (1977) review the results of the various preclinical and clinical studies

  using 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 investigators through the National Cancer Institute's Investigational Drug Branch of the Cancer Therapy Evaluation Program, is supplied as a freeze-dried white powder in vials containing 10 mg of cisplatin, 90 mg of sodium chloride, 100 mg of mannitol (USP) and

  hydrochloric acid to adjust the pH. When the product is

  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 NaCl.

  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 regarding the use of cisplatin in the treatment of 65 human patients

  having a variety of neoplasms. As in the previous publication

  dente, the drug is supplied to them by the National Cancer Institute in vials containing 10 mg of cisplatin, 90 mg of sodium chloride and mg of mannitol, so that the product can be reconstituted with 10 ml

of sterile water.

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

in 8 to 10 ml of sterile water.

  Some information about chemistry and pharmaceutical formulation

  of cisplatin are provided in pages 1 to 5 and 31 to 32 of

  the publication entitled: "CLINICAL BROCHURE, CIS-PLATINUM (II) DIAMINE-

  DICHLORIDE (NSC-119875), H. Haldelsman et al., "Investigational Drug Branch, Cancer Chemotherapy Evaluation Program, Division of Cancer Treatment,

  National Cancer Institute (Revised August 1974).

  On pages 31 and 32 of the latter concerning the formulation of cisplatin supplied gratis by N.C.I. clinicians to conduct their clinical evaluation in cancer chemotherapy, the following indications are made:

PHARMACEUTICAL DATA SHEET.

  NSC-119875 - cis-diaminedichloroplatin (11).

  Dose formulation mg / vial Preparation of solui mg / vial Storage Stability: The contents of each 20ml lead glass vial resemble a white lyophilized cake Each vial contains 10 mg of NSC-119875, 90 mg of sodium chloride , 100 mg of mannitol and

  hydrochloric acid to adjust ph.

  When the product is reconstituted with 10 mg of water

  sterile for injection, USP, each ml of the solution

  resultant contains 1 mg of NSC-119875, mg of mannitol and 9 mg of sodium chloride

  having a pH of between 3.5 and 4.5.

  : Unopened dry flasks must be stored

  at refrigeration temperatures (4 to 8 C).

  : The intact vials have provisional stability

  year when stored at a time

  refrigeration temperature (4 to 8 C).

  Stability recommendations can be adjusted

  pending the completion of a study of a

shelf life of two years.

  Reconstitution as indicated provides a pale yellow solution that is stable for up to 1 hour at room temperature

  (22 C) when exposed to ambient lighting

  normal binder and at a maximum of 8 hours at room temperature (22 C) when protected from light. The reconstituted solutions can form a precipitate, after 1 hour, at the temperature

refrigeration (4 to 80C).

  Caution The freeze-dried dose formulations do not contain any preservatives and it is therefore desirable to reject the solutions 8 hours after reconstitution. Published British Patent Application No. 2021946A discloses solutions

  stable aqueous levels of cisplatin with a concentration of cisplatin

  taken between about 0.1 and 1.0 mg / ml and a pH between 2.0 and 3.0.

  The solutions may also contain a pharmaceutically acceptable and non-toxic source of inorganic chloride ions, such as

  sodium chloride, and an excipient such as mannitol.

  The present invention relates to concentrated stable solutions

  cisplatin with concentrations 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 about 30 to 95% polyethylene glycol, having an average molecular weight of about

  and 9000, or a methoxy polyethylene glycol having a molecular weight of

  average of between about 300 and 6000, or a mixture thereof, and about 5 to 70% water, these solutions also containing at least one

  2O source of pharmaceutically acceptable and non-pharmaceutically acceptable chloride ion

  toxic, in an amount that is at least, approximately equivalent to the amount

  of cisplatin present in the solution, and these solutions having a concentration

  in cisplatin between about 2.5 and 25 mg / ml.

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

  cisplatin concentrations well above about 0.5 mg / ml.

  It is not easy to redissolve this crystalline cisplatin by shaking the system at room temperature, although a solution can be obtained again by heating to about 37 C. As the transport temperatures

  and after-sales storage can not be controlled, and that

  cisplatin in the vials would cause problems

  the doctor who uses it, the maximum practical concentration

  The amount of cisplatin in such aqueous media is about 0.5 mg / ml.

  The cisplatin solutions according to the invention may contain up to

  than about 25 mg 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 40C for 12 hours.

  month without observing a crystallization of cisplatin.

  Such solutions have also been frozen at -60'C, then thawed at

  room temperature without observing precipitation of cisplatin.

  Thus, practical solutions prepared according to the invention can offer-

  cisplatin concentrations at least 30 times higher than those

  may have practical aqueous solutions prepared according to the prior art.

  laughing. It will be appreciated that the concentrated solutions of cisplatin provided according to the present invention require lower storage and other 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 the

philisation.

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

  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 about 90% of polyethylene glycol, having a molecular weight

  average between 350 and 450, and about 10% water.

  It is preferable that the solution contains about 5 to 20 mg of

  cisplatin per ml, and still more preferably about 10 to 15 mg / ml.

  The source of pharmaceutically acceptable and non-pharmaceutically acceptable chloride ion

  toxic is preferably present at the concentration of at least about

  2 equivalents per equivalent of cisplatin in the solution. Concentrations

  as high as 50 equivalents or more of chloride ion per

  equivalent of cisplatin can be used, depending on the concentration

  of cisplatin, the percentage of water present and the particular source

  chloride ion, but such high chlorine ion concentrations

  are generally neither necessary nor desirable.

  It will be noted by those skilled in the art that, in the case of a strong

  It would not be possible to dissolve a sufficient amount of a chloride ion source such as sodium chloride to provide 50 equivalents of chloride ion per equivalent of cisplatin. In addition, a saturated or substantially saturated solution of a mineral chloride would be undesirable since

  crystallize in the solution when it is cold.

  In the situation described above, 50 equivalents of chloride ion per equivalent

  The slow release of cisplatin could be achieved with hydrochloric acid as a source of chloride ion, but this could provide a solution with

  an excessively high acidity, that is to say a low pH.

  The Applicant has found that excessively acidic solutions are

  somewhat less stable than more moderately acidic solutions.

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

  7 equivalents of chloride ion per equivalent of cisplatin.

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

of these.

  The polyethylene glycols and methoxy polyethylene glycols have the following general formulas: H (OCH 2 CH 2) n OH and CH 3 (OCH 2 CH 2) nOCH 3 respectively, and are available industrially under the names

  CARBOWAe polyethylene glycols and methoxy polyethylene glycols CARBOWA) È.

  The Applicant prefers to use the SENTRY qualities of CARBOWAX polyethylene glycols, which are produced to meet U.S.P. specifications.

  N.F. and F.C.C. concerning food and drug applications.

  The typical molecular weight ranges for a variety of CARBOWAX polyethylene glycols and CARBOWAX methoxy polyethylene glycols are provided

in Tables I and II, below.

TABLE I

  Polyethylene Typical Range CARBOWAX Glycols Molecular Weight

190 to 210

300,285 to 315

400 380 to 420

600 570 to 630

1000 950 to 1050

1540 1300 to 1600

4000 3000 to 3700

6000 7000 to 9000

TABLE II

  Methoxy Polyethylene Typical Range Molecular Weight Glycols

CARBOWAX

350,335 to 365

550,525 to 575

750,715 to 785

2000 1850 to 2150

  It is known that polyethylene glycols form complexes with certain mineral salts. Thus, the reaction of polyethylene glycols with blue complex ammonium cobalt thiocyanate forms the basis of a colorimetric method for determining the concentration of polyethylene glycols in various mixtures. The Applicant believes 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

  falls within the theory and does not constitute an element of the invention.

  Evidence of complex formation was not noted in the chroma-

  thin layer graphs made by the Applicant.

  Using the techniques described hereinafter, aqueous cisplatin produces a stain for an Rf value of about 0.65. However, a solution of cisplatin in 90% polyethylene glycol 400 - 10% water 10% HCl at 0.5N) provides a stain for an Rf of about 0.3.

  spreads to Rf of about 0.65. Ladilution with significant quantities

  water seems to break the complex immediately, since dilution of the above solution of PEG-H20 (or HC1) with five volumes of water then shows only a spot corresponding to cisplatin for a Rf

about 0.65.

  Thin layer chromatography (TLC)

Apparatus and reagents -

  (a) TLC plates: 60 silica gel plates from Laboratoires EM,

Catalog No. 5753, or equivalent.

  (b) Eluent - acetone:> Normal nitric acid (9/1). Prepared frankly

everyday.

  (c) Developer: Dissolve 5.6 g of stannous chloride in 10 ml of concentrated hydrochloric acid. Add 90 ml of distilled water and 0.29 of potassium iodide. Shake well. Prepare a fresh product

everyday.

  (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].

  (b) A TLC plate is stained with 5 microliters of the sample and microliters of a standard solution containing cisplatin (and transplatin 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

  pre-balanced CCM tank with eluent.

  Dried plaque will be sprayed with the developer solution freshly

  prepared and introduced into an oven at 1000C for 10 minutes.

  We observe the yellow / purple zones.

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

  HPLC titrations of the solutions according to the invention tending to determine

  the cisplatin content can be achieved according to the procedure

  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 concentration of cisplatin of 1 mg / ml.

  Although their presence does not confer any particular advantage, the solutions according to the invention may optionally contain an excipient

  Acceptable classical physiological point of view such as mannitol.

  From the stability studies conducted at present, it is expected that the stability of the solutions according to the invention (as being a loss 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 free of pyrogenics. Such solutions

  can then be diluted with, for example, sterile water for injection

  quality U.S.P. grade, or sterile, normal saline

  U.S.P., and administered intramuscularly or intravenously.

  The means for sterilizing such solutions are well known in the art.

  The Applicant prefers to pass the solutions through a filter -

  millipore of 0.22 micron, sterile and devoid of pyrogenic substance,

  using aseptic techniques under sterile nitrogen pressure.

  Millipore is a trade name of Millipore Corporation for membrane filters. The sterile filtrate is collected in sterile, pyrogen-free containers and finally introduced in the desired amount into suitable sterile pyrogen-free containers capped with sterile, pyrogen-free corks (preferably coated with Teflon). and

  hermetically closed with sterile aluminum capsules.

  For use in the treatment of cancer, the concentrated solutions are diluted to the desired concentration (usually 1 mg of cisplatin per ml) with for example sterile water for injection, USP quality, sterile normal saline, quality USP, or a sterile glucose solution, used by intravenous or intramuscular injection, or by intravenous infusion, in the manner known to date for

  administration of cisplatin preparations.

  Doses currently used with mild to moderate toxicity

  acceptable, range from 60 to 100 mg / m2 intravenously, administered in a single dose, or spread over 3 to 5 days, this dose being re-administered at 4-week intervals. A dose of 60 mg / m2 is approximately equivalent to a dose of 1.5 mg / kg which, in turn, is

  equivalent to a dose of 105 mg for a patient weighing 70 kg.

  Concurrent therapy with other chemotherapeutic agents is frequently prescribed.

  therapies to achieve the best results.

  As used herein in the claims, references to

  "equivalents" of chloride ion per "equivalent" of cisplatin mean molar equivalents. Thus, for example, when the preferred range of about 3 to 7 equivalents of chloride ion per equivalent of cisplatin is used, about 3 to 7 moles of NaCl per mole of cisplatin would be used but about 1.5 to about 3, 5 moles of CaCl 2 per mole of cisplatin, etc. Platinum B is an arbitrary designation used herein for the cisplatin acid reaction product which is half of the known red Magnus complex and to which the following structure has been attempted: e-CLe F \ Pt / l

LH3N / \

  This invention is illustrated but can not be limited by

Following examples.

EXAMPLE 1

  Stable concentrated solution of cispLatin (10 mg / ml) in 90% polyethylene

glycol 400 - 10% normal HCl -

  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 stirring for 2 days at room temperature (the container being protected from light by means of an aluminum foil), a solution is obtained.

light yellow.

2480605S

  Aliquots of this solution are added to 17 ml amber flasks, sealed with Teflon-coated stoppers, sealed with aluminum capsules and tested for storage stability at various temperatures. After two weeks storage at 56% C, thin layer chromatography (TLC) indicates the presence of a trace transplatin and about 1% platinum B. After storage for two weeks at 560C, the TLC indicates the presence less than 1% transplatin and about 3% platinum B. The samples stored for two weeks at 560C, are diluted with, respectively, 4, 9 and 19 volumes of water, and provide solutions

  transparencies containing, respectively, 2, 1 and 0.5 mg / ml of cisplatin.

  A slight cloudiness appears in each of the diluted samples after a

  about 18 hours at room temperature.

EXAMPLE 2

  * Stable concentrated solution of cisplatin (10 mg / ml) in 90% polyethylene

glycol 400 - 10% 0.5N HCl

  ml of U.S.P. grade purified water 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

  protected from light with aluminum foil and stirred at

  ambient temperature, for 24 hours, so as 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 added to 17 ml amber vials, sealed with teflon-coated caps and sealed with aluminum capsules, and then subjected to storage stability tests.

various temperatures.

  A sample vial is lyophilized in a dry ice bath.

  acetone for 1 hour, then allowed to warm to room temperature.

  A clear solution is obtained, and the presence of precipitate is not observed.

  After 3 months storage at 37% C and 45% C, the CCM indicates the presence of

  1 to 2% platinum B and a possible trace of transplatin.

  After 1 month of storage at 56% C, the CCM indicates the presence of more than 5% but

  less than 10% platinum B and a trace of transplatin.

  Samples stored at 37 and 450C for 3 months, and at 560C for 1 month, are diluted with four volumes of U.S.P. grade purified water. for

  give clear solutions containing 2 mg / ml of cisplatin.

  The diluted solutions remain clear after a rest of 24 hours at room temperature.

EXAMPLE 3

  stable stable solution of cisplatin (10 mg / ml) + CaCl2 (20 mg / ml)

  in 90% polyethylene glycol 400 - 10% 0.5N HCl -

  2 g of anhydrous calcium chloride of reactive quality 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 by means of aluminum foil; the system is stirred at ambient temperature for 24 hours to obtain a solution

clear.

  The TLC of a freshly prepared solution indicates only the presence

  of a cisplatin zone with a possible trace of transplatin.

  2 ml aliquots of the solution are placed in 17 ml amber vials, sealed with Teflon-coated stoppers, sealed with aluminum capsules and subjected to storage stability tests.

various temperatures.

  A sample vial is placed in a dry ice-acetone bath for 0.5 hours and lyophilized to a clear gel. We leave this one

  warm to room temperature and collect a clear solution.

* After storage for 3 months at 37 ° C, the CCM indicates the presence of 1 to 2% of

  platinum B and a possible trace of transplatin.

  After a storage period of 3 months at 45 C, the CCM indicates the presence of about 5%

  platinum B and a possible trace of transplatin.

  After a storage period of 1 month at 56 C, the CCM indicates the presence of 8 to 10%

  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 U.S.P. for

  give clear solutions containing 2 mg / ml of cisplatin.

  Diluted solutions remain clear after a 24-hour rest at

ambient 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 -

  3 ml of a solution of 90% of polyethylene glycol 400 and 10% of 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.

13 2480605

  An additional dose of 30 mg of cisplatin (25 mg / ml total) is added and the mixture is stirred at about 45 ° C for 1 hour, then at room temperature.

  room temperature for 16 hours to produce a practical solution.

  completely. The small amount of insoluble material is removed by filtration. The TLC of the filtrate indicates only one area of cisplatin with a possible trace of transplatin. The remainder of the filtrate is introduced into a 17 ml amber vial, sealed with a Teflon-coated stopper, sealed

  with an aluminum capsule and kept at 45 C for 3 months.

  After an aging of 3 months at 45 C, the CCM indicates that there is 1 to 2% of

  platinum B and no transplatin in the solution.

  A dilution of the aged solution with U.S.P. grade purified water.

  at a concentration of 2 mg / ml of cisplatin provides clear solutions,

  which remain after a rest at room temperature for 24 hours.

EXAMPLE 5

  * Stable concentrated solution of cisplatin (12 mg / ml) + NaCl (10 mg / ml)

  in 90% polyethylene glycol 400 - 10% 0.5N HCl

  100 mg of sodium chloride are dissolved in a solution of 5 ml of U.S.P. grade purified water. and 5 ml of normal hydrochloric acid. This solution is supplemented with 90 ml of polyethylene glycol 400 and the mixture is stirred for 15 minutes. 50 ml of the solution obtained are added

  500 mg of cisplatin, and the mixture is shaken in the dark at the same time.

  room temperature for 3 days, to produce a clear solution.

  The TLC of the freshly prepared solution shows only a corresponding stain.

  laying in cisplatin. Titration by high performance liquid chromatography

  (HPLC) of the freshly prepared solution indicates that the solution contains

12 mg of cisplatin per ml.

  Aliquots are introduced into 17 ml amber flasks, the latter being sealed as described in Example 3, and subject to the requirements of

  storage stability at various temperatures.

  After storage at 37 C for 3 months, the CCM indicates that there is less

  1% platinum B and no transplatin.

  After storage at 56 C for 1 month, the CCM indicates that there is 1 to 2% of

platinum B and no transplatin.

  Dilution of aged samples with U.S.P. purified water up to a concentration of 2 mg / ml of cisplatin provides clear solutions

  which remain after a rest 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 losses of

  power of 6.6%, 6.1% and 2.3%, respectively.

14 2480605

EXAMPLE 6

  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 U.S.P. grade purified water. 45 ml of polyethylene glycol 400 are added with 500 mg of cisplatin and the mixture is stirred in the dark at room temperature for 6 hours.

to provide a clear solution.

  The TLC of the freshly prepared solution only reveals the presence of the spot corresponding to cisplatin; HPLC titration indicates that this

  solution contains 11.4 mg of cisplatin per ml.

  Aliquots are added to 17 ml amber flasks and sealed as described in Example 3, and then subjected to

  storage stability at various temperatures.

  After storage at 37 C and 45 C for 3 months, the CCM indicates that there is 1%

  platinum B and no transplatin.

  After storage at 56 C for 1 month, the CCM indicates that there is 2 to 3% of

platinum B and no transplatin.

  Dilution of aged samples with U.S.P. grade purified water.

  at a concentration of 2 mg per ml of cisplatin provides clear solutions

  which remain after a rest at room temperature for 24 hours.

  HPLC titrations of samples stored for 3 months at 45 ° C., 6 months at 37 ° C. and 7 months at room temperature show power losses of 6.1%.

7.0 and 0.9%, respectively.

EXAMPLE 7

  * 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 U.S.P. grade, 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 CCM of the positively prepared solution indicates only a corresponding spot

to cisplatin.

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

ambient or at 4 C.

  Aliquots of the positively prepared solution are introduced into 17 ml amber vials, which are sealed as described in

15.2480605

  EXAMPLE 3, and then subjected to storage stability tests at various times.

  tures. After storage at 37 C and 45 C, for 3 months, the CCM indicates that there is

  1% platinum B and no transplatin.

  After storage at 56 C for 1 month, the CCM indicates that there is 3.4% of

platinum B and no transplatin.

  Dilution of aged samples with U. S.P. grade purified water

  at a concentration of 2 mg / ml of cisplatin provides clear solutions,

  which remain after resting at room temperature for 24 hours.

EXAMPLE 8

  Concentrated solution of cisplatin (10 mg / ml) + NaC [(10 mg / ml)

  in 90% polyethylene glycol 400 - 10% 0.2N HCl -

  A solution of 0.5 g of NaCl in 4 ml of USP grade purified water, 1 ml of normal hydrochloric acid and 45 ml of polyethylene glycol 400 is added with 250 mg of cisplatin, and the mixture is shaken in the dark. at room temperature, for 4 hours, so as 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 after standing at room temperature for 24 hours. Dilutions with 1, 2, 3, 4 and 5 volumes do not present

  no crystallizations when maintained 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 1% of

  platinum B and a possible trace of transplatin.

  After storage at 45 C, for 3 months, the CCM indicates that there is 5% of

  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 transplatin.

  Dilution of aged samples with U. S.P. grade purified water

  at a concentration of 2 mg / ml of cisplatin, provides clear solutions,

  which remain after a rest for 24 hours at room temperature.

16 2480605

EXAMPLE 9

  * Stable concentrated solution of cisplatin (2.5 mg / ml), NaCl (9 mg / ml) and mannitol (12.5 mg / ml) in 31% (weight / volume) of polyethylene glycol

Acidified aqueous 6000 -

  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 U.S.P. grade. 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 are added

  tine and stir the mixture in the dark for 3 days at room temperature

  ambient to obtain a clear solution.

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

with cisplatin.

  Aliquots of the freshly prepared solution are added to 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 CCM indicates that there is less

  1% platinum B and no transplatin.

  Dilution of aged samples with an equal volume of U.S.P. grade purified water. provides clear solutions, which remain so after a

  rest at room temperature for 24 hours.

EXAMPLE 10

  * Stable concentrated solution of cisplatin (15.8 mg / ml) + NaCl (10 mg / ml)

  in '90% of aqueous polyethylene glycol 400 -

  90 ml of polyethylene glycol 400 are dissolved in a solution of 1.0 g of NaCl in 10 ml of U.S.P. grade purified water. 60 ml of the solution obtained are added with 900 mg of cisplatin, 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 area corresponding to cisplatin; a HPLC titration indicates that it

  contains 15.8 mg of cisplatin per ml.

  Dilution of the freshly prepared solution with 4 volumes of U.S.P. grade purified water. provides a clear solution, which remain so after a

  rest at room temperature for 24 hours.

  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 2 months of storage at 45 C, the CCM indicates that there is 1.5% of platinum B

and no transplatin.

  After 1 month of storage at 56 C, the CCM indicates that there is 2% platinum B

and no transplatin.

  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

  stay after resting at room temperature for 24 hours.

  HPLC titrations of samples stored at 45 C for 3 months, at 37 C for 6 months and at room temperature for 8 months indicate that

7.6%, 7.6% and 0%, respectively.

EXAMPLE 11

  * 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 CaCl 2 in 10 ml of U.S.P. grade purified water.

  is added 90 ml of polyethylene glycol 400 and the resulting solution is stirred for 10 min. To 50 ml of the above solution, 750 mg of cisplatin are added and the mixture is stirred for 5 hours in the dark, at room temperature.

  room temperature, to obtain a clear solution.

  The TLC of the freshly prepared solution indicates only a correct zone

  responding to cisplatin. Dilutions of the freshly prepared solution

  with 1, 2, 5 and 10 volumes, respectively, of purified water of U.S. P. quality.

provide a clear solution.

  Aliquots of 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 a storage period of 2 months at 45 C, the TLC indicates that there is 1.5% platinum B

and no transplatin.

  After a storage period of 1 month at 56 C, the CCM indicates that there is 2.5 to 5% of

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 so after a rest at

  room temperature for 24 hours.

EXAMPLE 12

  * Stable stable concentrated solution of cispLatin in 90% polyethylenyl 400 - 10% 0.5N HCl (label indicates 15 mg / ml of activity

cisplatin) -

  NOTE: Cisplatin is a possible carcinogenic agent. You have to wear clothes, gloves, mask, glasses and hat

  protection throughout the procedure.

  All work areas and equipment must be

  thoroughly cleaned to prevent future contamination.

FORMULA

  per ml per 10.0 ml Cisplatin ............... 0.015 g 0.150 g Sodium chloride ..... 0.015 g 0.150 g Hydrochloric acid 0.5N 2) ........ ... 0.10 ml 1.0 ml Polyethylene glycol 400 (SENTRY quality) ... qsp 1.0 ml q.s. 10.0 ml

  1) This weight of cisplatin provides a power of 1000 micrograms / mg.

  To determine the amount of cisplatin required, the following formula is used: 1000 x 0.015g = Grams of cisplatin required power of cisplatin in mcg / mg 2) 1 liter of 0.5N hydrochloric acid is prepared as follows: 1. 957.25 ml of sterile water for injection, quality

U.S.P. in an Erlenmeyer flask.

  2. With rapid stirring, 42.75 ml of concentrated hydrochloric acid are slowly and cautiously introduced. It is stirred again during

  min. Close with a clean butyl rubber stopper.

q

  MANUFACTURING INSTRUCTIONS FOR PREPARING A LITER OF STERILE SOLUTION.

  1- Place 100 ml of 0.5N hydrochloric acid in a calibrated Erlenmeyer flask of 1U, containing a suitable stirrer such as a bar

  6 cm magnetic stirrer coated with teflon.

  15 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 minutes. 4 - We remove the magnetic bar and we drain it so that the

  excess fluid falls back into the falcon.

  15 g of cisplatin activity are carefully added.

  b - Polyethylene glycol 400 (SENTRY quality) is added to the brand

  1k (A total of 880 ml of polyethylene glycol 400 is required).

  7 - Teflon stirrer is put back into the mix and the system is closed

  with a clean stopper of butyl rubber.

  a - The container is wrapped with aluminum foil to protect it

light.

  q - Stir rapidly for 24 to 48 hours at room temperature.

  A clear solution must be obtained. If a clear solution is not obtained in 48 hours, the mixture can be heated at 37.degree.-400.degree. C. for 2 to 6 hours, in the absence of air and light, to facilitate dissolution.

  rapid cisplatin remaining. It is cooled to 23-270C.

  Using an aseptic technique, the dark yellow solution is passed under a suitable sterile nitrogen pressure through a sterile, pyrogen-free, 0.22 micron Millipore filter. The sterile filtrate is collected in a sterile Erlenmeyer flask free of pyrogen. We close the flask with a cap

  sterile, pyrogen-free, butyl rubber.

  The solution must be kept in the dark.

  The necessary amount of sterile solution is introduced into sterile amber vials devoid of pyrogen. These vials are closed with a sterile Teflon stopper and free of pyrogen. These vials are sealed with capsules

sterile aluminum.

  12 - The vials must contain the following warning label:

  PRODUCT NOT TO BE USED DIRECTLY INTRAMUSCULARLY

OR INTRAVENOUS *

  * The PEG li400 solution can be diluted with 14 parts U.S.P. sterile water for injection. or sterile normal saline

  quality U.S.P. to provide a solution of 1 mg / ml of cisplatin.

  If higher concentrations are required, quan-

  proportionately less sterile water or saline solution.

  The diluted solutions can be used intravenously and are stable at room temperature (22 to 26 C) for at least 48 hours. Do not put diluted solutions in the refrigerator

  because crystals can appear.

  13 - Keep the vials in the dark.

EXAMPLE 13

  e Stable concentrated solution of cisplatin (2.5 mg / ml), NaCl (9 mg / ml), CaCl2 (15 mg / ml) and mannitol (10 mg / ml) in 31% polyethylene glycol

Acidified aqueous 400 -

  0.9 g of sodium chloride, 1.5 g of CaCl 2 and 1.0 g of mannitol are dissolved in a mixture of 31.25 ml of polyethylene glycol 400 and 40 ml of purified water of U.S.P. grade. The solution is acidified to pH 2.2 with normal hydrochloric acid (0.4 ml), then 255 mg of cisplatin is added and the volume is brought to 100 ml with USP grade purified water. ; the mixture is stirred in the dark for 5 hours at room temperature

  ambient, to obtain a clear solution.

  The TLC of the freshly prepared solution indicates only a corresponding zone.

with 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; we thus obtain a concentrated solution

stable cisplatin.

EXAMPLE 16

  The general procedure described in Expt 5 is repeated, except that polyethylene glycol 400 is replaced by an equal volume of polyethylene

  glycols 200 and 300, respectively. Static concentrated solutions are obtained

of cisplatin.

EXAMPLE 17

  We repeat the general procedure described dance 'Exempt 9, if it is not

  polyethylene glycol 6000 is replaced by an equal weight of polyethylene

  1000 and 4000 glycols respectively. Concentrated solutions are

stable tracts of cisplatin.

  Of course, the present invention is not limited to the examples

  and modes of implementation mentioned above; it is likely to

  Many variants accessible to those skilled in the art, depending on the applications

  contemplated and without departing from the spirit of the invention.

Claims (8)

  1.- stable concentrated solution of cisplatin in a solvent medium, characterized in that it comprises about 30 to 95% of polyethylene glycol having an average molecular weight of between about 150 and 9000, or a methoxy polyethylene glycol having an average molecular weight between about 300 and 6000, or a mixture thereof, and about 5 to 70% water, and also at least one non-toxic and pharmaceutically acceptable source of non-toxic chloride ion, an amount that is at least about equivalent to the amount of cisplatin present in the solution, which solution has a concentration of cisplatin included between about 2.5 and 25 mg / ml.   2.- stable concentrated solution of cisplatin in a solvent medium, characterized in that it comprises about 80 to 95% of polyethylene glycol having a mean molecular weight of between about 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 to 20% water, as well as at least one non-toxic and pharmaceutically acceptable source of chloride ion, in an amount of which is between about one equivalent and ten equivalents per equivalent of cisplatin to the solution, this solution containing about 5 to 20 mg of cisplatin per ml.   3.- cisplatin solution according to claim 2, characterized in that the source of non-toxic chloride ion and pharmaceutically acceptable is constituted by hydrochloric acid, sodium chloride, calcium chloride, magnesium, or triethylamine hydrochloride, or a mixture of these.   4.- stable concentrated solution of cisplatin in a solvent medium, characterized in that it comprises about 85 to 90% of polyethylene glycol having a mean molecular weight of between about 250 and 650, and about 10 to 15% of water, as well as additionally a chloride ion source selected from the group consisting of hydrochloric acid, sodium chloride or their mixtures, in an amount which is between about 2 and 7 equivalents per equivalent of cisplatin in the solution, this solution   having a cisplatin concentration of between about 10 and 15 mlg / ml.   5.- cisplatin solution according to claim 4, characterized in that the solution is sterile and is in a closed container hermetically, such as a vial. 23 2480605   6. Sterile stable concentrate solution of cisplatin in a hermetically sealed container such as a vial, characterized in that it contains about 10 to 15 mg of cisplatin per ml, and about 10 to 15 mg of NaCl per ml in a medium solvent comprising about 90% of a polyethylene glycol having an average molecular weight between about 350 and 450, and about % of water.   7. Sterile stable concentrate solution of cisplatin in a sealed container such as a vial, characterized in that it contains about 10 to mg of cisplatin per ml, and about 10 to 15 mg of NaCl per ml in a solvent medium consisting of by about 90% of a polyethylene glycol having an average molecular weight between about 350 and 450, and about% diluted hydrochloric acid, whose concentration can be up to 0.5N. 8.- Pharmacological application of stable concentrated solutions   according to any one of claims 1 to 7, in particular for the therapy cancer. @