CZ278891B6 - Medicinal preparation of cis-platinum and process for preparing thereof - Google Patents

Abstract

The invention concerns an anti-tumour medicinal preparation whose active substance is cis-diaminedichloroplatinum complex (cis-platinum, cis-DDP) and as adjuvants potassium, ammonium and sodium chloride, which in a certain ratio significantly increase the stability of cis-platinum.

Description

Cisplatin antitumor medicinal product and process for its preparation

Technical field

The present invention relates to an antitumor medicinal product, the main active ingredient of which is the coordinating compound cis-diamminedichloroplatinum (II) complex with the international designation cisplatin or cis-DDP.

BACKGROUND OF THE INVENTION

In the field of human medicine, this platinum cytostatic drug was introduced in 1979 in the form of a lyophilized injection containing 9 parts by weight, parts by weight of sodium chloride and 10 parts by weight, parts by weight of mannitol per part by weight, part by weight of cisplatin. Dry injection after addition of the prescribed amount of water provides a solution with a pH of about 4-6 (Ann. Int.Med. 86, 803-812, 1977).

Various studies published in the literature have later shown that the presence of mannitol in the formulations is not beneficial and rather reduces their stability.

However, the presence of chloride ions in cisplatin dosage forms is necessary to stabilize the electroneutral but highly water-reactive cis-DDP complex, which can form a number of charged and, at higher pH, electroneutral hydrolysis products, including binuclear complexes (J.Am.Chem.Soc 83: 2457-2462 (1961; J. Am. Chem. Soc. 99, 771-781, 1977). It is well known that these hydrolytic products of cisplatin are highly toxic and virtually ineffective in antitumor, so that their formation in injection is highly undesirable (Cancer Treat.Rep. 63, 1433-1438, 1979).

Use of sodium chloride as excipient injections containing cisplatin, thus limiting terms ligand substitution reaction ^of Cl ^- ligands of water entirely justified, which is used in some commercial preparations - of aqueous solution composed of:

cisplatin 0.5 mg / ml sodium chloride 9.0 mg / ml hydrogen chloride and pH = 2.0 to 3.0

This pharmaceutical formulation is the subject of patent protection, the basis of which is based on the finding of the authors that said narrow pH range favorably affects the stability of the active substance in solution, the most optimal being considered to be pH = 2.5. High Performance Liquid Chromatography (HPLC) showed that

The stability of the liquid injections of the above composition is good, and they lose effectiveness during 3-4 months of storage at 56 ° C, i.e. the cis-DDP content decreases by only a few percent (US Patent 4310515).

We performed a comprehensive study aimed at monitoring the substitution reactions of cisplatin when using more sensitive identification of products of the reactions taking place, however, show that in a strongly acidic pH, there is a significant extent to the substitution of a ligand NH ₃ ligand Cl "forming ammintrichloroplatnatého complex (ATC). We have also found that the proportion of individual substitution reactions depends not only on the acidity of the solution, but also on the composition of the cations added in the form of chlorides.

Potassium cation is more preferred than Na ^{+ for} the formation of the ATC complex. Even less of this complex is formed in NH ₄ Cl solutions, even at elevated temperatures.

Although the presence of NH ⁺ ₄ in cisplatin solutions results in the formation of triamminchloroplatinum (TAC) due to ammination, this is a much more acceptable additive in terms of effects at the molecular level (the target of cisplatin attack is DNA), since unlike the trivalent ATC to biomacromolecules of organism only one point and therefore its overall biological effect is negligible. Therefore, only the content of the ATC complex is limited in the most recent edition of the US Pharmacopoeia (USP XXII).

SUMMARY OF THE INVENTION

Based on the above findings, the stability of various formulations of cisplatin in aqueous solutions containing ammonium chloride and a combination of other alkali and alkaline earth chlorides has been studied for a long time. The result of the stability study is a new medicinal product cisplatin in aqueous chloride ion solution, characterized in that the injection solution contains K ⁺ , NH ⁺ ₄ , Na ⁺ as a cation in a molar ratio from 1:10:10 to 1 : 30: 50 parts and H ⁺ , corresponding to a pH of 3 to 4.5 at a total chloride ion concentration of 90 to 420 mmol / l.

Preferably, the solution for injection contains 0.50 mg of cisplatin, 0.1 to 0.5 mg of potassium chloride, 1.5 to 4.5 mg of ammonium chloride, 2.5 to 6.5 mg of sodium chloride, and hydrogen chloride for treatment in 1 ml. pH to 3.1 to 3.5. An injection of this composition is prepared by dissolving potassium, ammonium and sodium chloride in water for injection, adjusting the pH of the solution to the desired value by adding hydrogen chloride, dissolving cisplatin at 20 ° C to 25 ° C, then the solution is sterilized by ultrafiltration, filled into individual vials under aseptic conditions, and these are sealed with a piercing plug with an inert inner surface and secured with an aluminum cap. If desired, this dosage form of cisplatin can also be adjusted into suitable bulk containers, similar to infusion solutions.

The following requirements were taken into account when optimizing the ratio of individual chlorides in solution:

-2GB 278891 B6

- the solution for injection should be isotonic,

- the concentration of K ⁺ ions must be physiological,

- the usual volume of acidifying solution administered during one cycle of cis-DDP therapy must not lead to life-threatening acidosis,

- the infusion rate of the cis-DDP solution does not exceed 500 ml / h,

- the permissible rate of introduction of NH ⁺ ₄ into the body is 0,1 mmol / kg / min.

The advantages of the new dosage form of cisplatin are evident in terms of the stability of the formulation and its pharmacological effects.

Table 1 shows the concentrations of the active substance cis-DDP in a NaCl solution and in a solution of a mixture of K ⁺ , NH ⁺ ₄ , Na ⁺ , stored at 5, 25, 40, 50 and 60 ° C for a long time.

Tab. 1

Solution Temperature (•C) Concentration of cis-DDP (μg / ml) Initial status 2 months 6 months 14 months 27 months 5 507 504 516 512 506 25 507 502 511 517 428 NaCl 40 507 492 513 477 348 50 507 470 465 333 280 60 507 446 352 3 07 157 5 512 507 512 509 516 25 512 506 509 509 513 KC1 40 512 502 500 501 464 nh ₄ ci 50 512 512 493 410 328 NaCl 60 512 422 377 313 203

The HPLC data indicated that the stability of the injections containing the chloride mixture was greater at all temperatures and times than for NaCl alone.

Computer analysis has shown that, from a kinetic point of view, the loss of active substance is subject to the pseudo-first order and a min. 4 years at a storage temperature of 10-20 ° C.

The new dosage form of cis-DDP can also be considered stable in terms of other quality parameters. The injections, stored at 5-50 ° C, were clear for at least 27 months (the absorbance of the solution at λ = 500 nm in a 1 cm cuvette did not exceed 0.008). The presence of NH ₄ Cl in the injections significantly stabilizes the pH of the solution. The difference in pH values of the injections stored for 27 months at 25 ° C and 5 ° C was only 0.13 units, which is practically within the precision of long-term follow-up of this parameter.

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Electron spectra (UV-VIS) in the 240-500 nm region remained virtually unchanged at 5 ° C and 25 ° C. At higher temperatures, absorbance in the low spectrum (246-247 nm) gradually increased, but its character was maintained after 27 months.

In accordance with the loss of active substance at elevated temperatures, the formation of three decomposition products ATC, TAC and trans-DDP was observed by chromatographic methods (TLC, HPLC). In terms of biological effects, their representation can be said to be more favorable than in the case of cis-DDP solutions in the presence of sodium chloride alone.

Unlike NaCl, in the case of NH? Cl Cl ions all "acidic as NH ⁺ ₄ are metabolised in the liver. Thus, in addition to stabilizing the injectable solution, ammonium chloride causes acidic chlorouresis in vivo, which may prove to be a protective effect against cisplatin nephrotoxicity. Experimental determination of acute toxicity of the new dosage form of cis-DDP compared to its solution in 0.9% NaCl confirmed the above assumption because the relative toxicity of the new dosage form reached 93.24% in female mice with a confidence limit of 88.28-98 , 58% and in male mice only 84.62% with a confidence limit of 81.25-87.53% with a trial error of 5.52 and 5.52%, respectively. 3.71%.

Direct evidence of reduced nephrotoxicity of the new cis-DDP dosage form was obtained in male Wistar rats.

Overall, the toxicity of the preparation was also positively reflected in the anti-tumor activity tests in DBA / 2-J mice transplanted with 10 ⁵ leukemia P 388 cells on day 0.

The mean survival of the treated animals was 262% for the new dosage form and 242% for the cis-DDP solution in 0.9% NaCl.

The safety of administration of a new carrier solution (KCl, NH ₄ Cl, NaCl) with and without cis-DDP was active. tested on experimental calves, when the animals received 5 times higher doses of K and NH ₄ ions than infused with human drug by infusion. Even under these conditions, there were no significant changes in the internal environment.

Preclinical testing results were also confirmed in clinical trials where no grade 3 toxicity was reported in 56 application cycles. The tolerability of the new dosage form of cisplatin appeared to be better than the comparative commercial lyophilized formulation, but the group of 15 patients cannot be considered statistically significant.

The invention is illustrated by the following non-limiting examples.

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Examples

Example 1

Medicinal product containing 0.5 mg cisplatin, 4.65 mg sodium chloride, 0.3 mg potassium chloride, 3.0 mg ammonium chloride and hydrogen chloride ad adjust.pH 3.3 ± 0.2 in 1 ml solution for injection.

446.4 g of sodium chloride, 28.7 g of potassium chloride, 288.0 g of ammonium chloride are dissolved under aseptic conditions in 90.00 kg of water for injections and the pH of the solution is adjusted by adding about 10 ml of hydrochloric acid. ( ^c HCl ⁼ 5 mol / L) to 3.1. 48.00 g of the finely crystalline cis-DDP substance are added to the chloride solution and the solution is stirred for 30 minutes. After complete dissolution of the drug, make up the total weight of the solution to 96.00 kg with water for injection. A check pH whose value should be 3.1 to 3.2, and controlling the concentration and spectral purity of the solution, characterized by absorbance in a 1 cm cell at 300 nm and a = 0.10 ± 0.215 and the ratio ^A 3OO / ^A 246 4 5. A suitable solution is sterilized by ultrafiltration and the aseptic conditions are filled into 20.6 ml class 1 hydrolytic class vials under aseptic conditions, sealed with a Teflon stopper and secured with an aluminum cap. 4,500 vials of 10 mg cisplatin are obtained.

Example 2

The medicament of Example 1 containing 50 mg cisplatin per vial.

The procedure of Example 1 is followed, but the sterile ultrafiltrate is filled into vials of nominal volume of 100 ml in batches of 102.0 ml to give 900 vials of the formulation containing 50 mg of cisplatin.

Example 3

DBA / 2-J mice were intraperitoneally transplanted with 10 ⁵ P388 leukemia tumor cells on day 0. The animals, which were kept as 10 controls, perished on average (22.9 ± 1.9) days, whereas The other 10-member groups treated with Example 1 administered a 10 mg dose of cis-DDP / kq body weight on day 1 significantly increased life expectancy, on average (60.0 ± 11.9) days. The increase in survival is 262%.

Example 4

CBA x BL mice in 10-male subjects received 11.0 intravenously in 10-member groups; 12.0; 13.0; 14.0; 15.0; 16.0; 17.0 and 17.5 mg of cis-DDP as a solution for injection prepared according to Example 2 and as a solution of cis-DDP in 0.9% NaCl (reference standard).

-5GB 278891 B6 Walla

3.71

1.

Relative toxicity of the product

84.62% of the comparative value of the present invention of the standard when the experiment failed

Claims (3)

PATENTOVÉ Claims The antitumor medicinal product chloride ions, recognizes cisplatin in aqueous solution, in that the injection solution contains, in addition to the effective NH ⁺ ₄ , Na ⁺ in a molar ratio of 1:10:10 and H ⁺ , corresponding to a pH of 3 to 4.5 tration of chloride ions 90 to 420 mmol / l. K ⁺ cations, up to 1:30:50 parts at total conc The antitumor medicinal product according to claim 1, characterized in that the solution of the injectable solution preferably contains 0.50 mg of cisplatin, 0.1 to 0.5 mg of potassium chloride, 1.5 to 4.5 mg of ammonium chloride, 2, 5 to 6.5 mg of sodium chloride and hydrogen chloride to adjust the pH to 3.1 to 3.5. Method for the preparation of an antitumor medicinal product according to claim 1 and / or 2, characterized in that potassium, ammonium and sodium chloride are dissolved in water for injection, the pH of the solution is adjusted to the desired value by adding hydrogen chloride at a temperature of 20 ° C to 25 ° Cisplatin is dissolved in the solution while stirring, then the solution is sterilized by ultrafiltration, filled under aseptic conditions into individual vials and sealed with a piercing plug with an inert inner surface and secured with an aluminum cap.