SK19594A3 - New method of preparation of lyophilizate of cetrorelix - Google Patents

Abstract

Disclosed is a production method for sterile cetrorelix lyophilisate, which consists in that the cetrorelix acetate is dissolved in 27 to 33 wt. % of acetic acid, the obtained solution is diluted by water for injection purposes, and then to such solution the structured retentive agent, selected from the group comprising hexites, particularly mannit, glucide, sorbite, D-sorbite, dulcitol, alite and idite, and urea and sodium chloride, is added, and the obtained solution is sterilically filtrated, filled in the syringes and lyophilisated.

Description

Cetrorelix is an acid amide-terminated decapeptide that uses the eco-acetate salt. The synthesis and some pharmacological effects are described in patent application EP 299 402.

It should be possible to administer the active substance subcutaneously at a dose of 0.1 to 20 mg. Aqueous decapeptide solutions are not stable, autoclaving in the final container is not possible. The peptide has a tendency to degrade when sterilized according to the pharmacopoeia. Therefore, in order to obtain an injectable solution, it would be necessary to develop the levophilisate.

However, the amount of active ingredient in the solution to be lyophilized is so small that, at low concentrations of active ingredient, after drying without adjuvants on the glass, the ampoule is only slightly airborne, which is discharged from the amphiol with the steam flow used for sterilization. Therefore, it is necessary to use a carcass forming agent which forms a stable cake. At high concentrations, this excipient can be dispensed with. Suitable skeleton-forming agents are: hexites, in particular mannitol, sorbitol such as L-sorbitol, dulcite, allite, altrite (e.g. L-altrite and L-altrite), idite (e.g. D-idite and L-idite) their optical active forms (D-forms or L-forms) as well as the corresponding race mint. In particular, mannitol such as D-mannitol, L-mannitol, L-mannitol, sorbitol and / or dulcite, and preferably D-mannitol thereof, are used. Mixtures of the hexites mentioned, for example mixtures of mannitol and sorbitol and / or dulcite, can also be used as hexites. Since dulcite is less soluble in water than, for example, vary, the content of dulcite in the aqueous solution should not exceed, for example, d 3% by weight. In contrast, mannitol and sorbitol can be mixed in all proportions.

In addition to hexites, other conventional pharmaceutical agents such as amino acids such as alanine glycine, lysine, phenylelanine, aspartic acid, elutaminic acid, leucine, lactose, polyvinylpyrrolidone, glucose, fructose, albumin and equivalent skeletal formers can be used. Furthermore, urea and sodium chloride can be used as scaffolding agents. The total amount of such substances in solution used for freeze-drying is, for example, 0 to 16.9 parts by weight, for example 0.1 to 7 parts by weight, based on 1 part by weight of cetrorelix. In the finished lyophilisate, the total amount of such excipients may be up to 16.9 parts by weight, based on one part by weight of hexite. In detail, the amount of such excipients is governed by the amount of hexite present, namely that the total amount of hexite and the like in the finished lyophilisate is no more than 17 parts by weight based on 1 part by weight of cetrorelix. Thus, if only 0.1 parts by weight are present in the lyophilisate, up to 16.9 parts by weight other excipients may be present; for example, δ, δ parts by weight of hexite are present, for example, the amount of other substances may be up to 8 p parts by weight, based on 1 part by weight of cet-3 rorelix.

However, during the development work on the lyophilisate, it was necessary to state that the active ingredient was administered in the form of a formulation. behavior very differently unpredictable. The first batches yielded good results, but it was soon found that sterile filtration caused difficulties and defective batches.

From the literature, for example, from Powell, M-F ₀ : Pharmaceutical P.esearch, 1258-1263 / 8/1991; Lathe, M .: Int. J. Peptide Protein Dog. 344-349 / 36/1990; Szejtli, J .: Pharmaceutical Technology International 16-22, 1991, it is known that oligopeptides, especially those with an acid amide end-functional group, show a tendency to gel. In sterile filtration, this can be seen in terms of filtration rates, often even the organoleptically increased viscosity of such solutions is found. A gel layer remains on the sterile filter. As a result, it is not possible to produce a drug with a precisely defined active substance content. -...... ·

Table 1 shows the different results of the first batches.

The active ingredient contents vary between $ 100 and £ 36.

Table 1: cetrorelixacetate

Batch batching content of active substance y «* 1 100 µg 100 2 500 µg 100 3 500 µg 90

- 4 Table 1 (cont.): Cetrorelixacetate

batch batching content of active substance% 4 500 ug 36 5 500 ug 100 6 500 ug 85 7 1 mg 80 8 1 mg 100 9 ' 2 mg 100 10 2 mg 80 11 2 mg 100

In order to avoid this gel formation, the following ingredients have been listed in the letter of the tour, which have been used experimentally:

Suitable organic solvents are, for example, acetonitrile, n-butenol, tertiary butanol, ethanol, isopropanol and benzyl alcohol, and buffer salts such as acetate buffer, citrate buffer, sodium chloride, sodium phosphate may be used. , yellow sodium, r.atrium bi carbonate, phosphate buffer, gusnidiacetate, urea. Also, polymers such as gelatins, polyethylene glycol 600, hydroxyethyl starches, polyvinylpyrrolidone, polyvinyl alcohol can be used. Also described is the addition of amino acids such as alaine, gylcin, lysine, phenylalanine, aspartic acid, glutamic acid, and leucitine.

The acids used were citric acid, caprylic acid, octanoic acid, hydrochloric acid, sulfuric acid and acetic acid. Physiologically harmless

- 5 surfactants, benzalkonium chloride, cetyl alcohol, gallic acid, lecithins, poylsorbates are available,

R R

Spans and Pluroni.cs.

Carbohydrates and cyclodextrins such as glucose, lactose, mannitol, sucrose, alpha, beta and gamma cyclodextrins, hydroxypropyl-alpha-cyclodextrins and hydroxypropyl-beta-cyclodextrins, hydroxyethylcyclodextrins, and methylcyclodextrins have also been used. These excipients have been tested as filtering aids to prevent gel formation.

However, a satisfactory solution to the problem has not been found. Only acidification with acetic acid has brought partial success. But even here it was necessary to reconcile itself with higher filtration losses. Surprisingly, it has been found that cetrorelix can dissolve well in 30% v / v sulfuric acid. This solution is then filled to a final concentration of 3% cetrorelix with water for injection and mannitol is added. Although it is described in the literature that the terminal amide group is readily hydrolyzed under acidic conditions, this has not been found with cetrorelix. The solutions produced by this method do not cause any filtration problems. Correct active ingredient contents have always been found. ......

The filtration rate reaches values that ensure satisfactory production rates. The general process of sterile lyophilization is described in Sucker, Luchs and Speiser (publisher) Pharmezeutische Technologie, 2nd edition 1991, Thieme-Verlag, Stuttgart-New York, not the wall. 537-559. Furthermore, the I.yophilization process described in LE-OS 37 35 614 is used.

- 6 Levophilisate is used to treat sterility in women. The method of therapy is to stimulate follicle maturation with a human gonadotropin from a menstrual bleeding break and then to ovulate with a dose of human choriongonadotropin. This ovulation aroused 32 hours later. The egg cells thus obtained are available for extracorporeal fertilization.

The lack of this antagonist therapy causes up to 10 follicles to mature in the stimulation phase. This increase in maturation of the follicle leads to peaks of LH hormone levels. These peaks result in the early stage of follicle maturation and early ovulation. This ovulation disorder occurs in about 25% of the cases treated and presents a disadvantage of a cycle that shows such an ovulation disorder for which egg cells are obtained and the entire treatment must be repeated for about 1 month.

Yet another drawback of conventional stimulatory treatment is the long treatment time of 4 weeks, which is necessary to achieve suppression. Agonists continue to show hyperstimulation syndrome in 1 to 2 cases, in which the follicle cells herypertrophy. The risk of hyperstimulation is particularly high in semi-ovarian ovaries. Hyperstimulation syndrome is a serious side effect that can lead to death cases.

The antegonist cetrorelix has been shown to have the following benefits in this treatment:

A treatment time of 5 days is sufficient for cetrorelix treatment to achieve total suppression. Hyperthermia syndrome

- 7 mulchations cannot occur. In addition, in the second treatment phase, the ovulation-inducing phase, HMG can be saved. This presents no insignificant advantages with respect to the cost of this in vitro fertilization treatment. Fertilization. in vitro is used, for example, in anomalies of the oviducts. For this treatment, the cycle should be accurately controlled and the ovulation time determined as accurately as possible. To date, this has been achieved to a limited extent, since the stimulation with HMG / MCG has often resulted in an ovulatory increase in LH too early or has not been sustained for long enough. However, the decisive challenge for the outcome of treatment is to prevent this too early increase so that the exact time of fertilization can be determined. This reduces the physical and physical burden on patients and optimally uses clinical logistics. In order to achieve this goal with great reliability, it is necessary to suppress endogenous hormone production (LF-FSH, estradiol) as perfectly as possible in order to stimulate follicle maturation simultaneously with the dose of exogenous gonadotropin (EMG / HCG) and to control hormone status at any time. Only when a sufficient number of follicles (4-6) having approximately the same degree of maturation has been reached is ovulation induced by a dose of ECG bolus injection.

By using an anagonist, the treatment of patients can be performed more successfully and more confidently.

A more recent field of use of the cetrorelix lyophilisate of the present invention is for use in the protection of male gonads. Male patients are pretreated with cetrorelix lyophilisate, and the activity of the gonads is promoted. Thus, other damaging causes, such as radiation treatment or cytostatic treatment, have very few or no possibilities at all to affect the sensitive tissue of the gonads.

Examples

Production example

About 1.5 liters of water for injection purposes are provided in a suitable glass container. Another glass container is filled with 210 g of water for injection and 91.17 g of acetic acid. The calculated amount of cetrorelixecetate (1.62-1.695 g, depending on the batch used) is dissolved in the produced $ 30 acetic acid with stirring. This solution is transferred to a glass container with 1.5 liters of water for injection, 82.2 g of mannitol are added, dissolved and made up to 3039 g with water for injection.

Process controls:

pH value 2.5 to 3.0 density: 1.009 to 1.017 g / cm ^J at 20 ° C refractive index: 1.227 - 1.340 at 440 nm and ° C

Sterilization of the solution is accomplished by filtration through a suitable membrane filter (0.2 µm pore size) under aseptic conditions. Discard 100 ml of the front fraction. The filters are sterilized by pressure steam. The freeze-dried cetrorelix solution is kept protected from recontamination.

The solution is immediately dispensed under aseptic conditions into a vial of Lin 2R colorless, hydrolytic class I and fitted with sterile freeze-dried stoppers. The required amount is 2.0 ml = 2.026 g.

ml vials were rinsed on the vial washing device and dried with hot air and sterilized. The cleaned freeze-dried stoppers were autoclaved. The pre-sealed vials were transferred to a freeze-dryer and allowed to freeze at -40 ° C. Drying is carried out by means of a drying program at a plate temperature from -40 ° C to + 20 ° C. The device is then filled with sterile nitrogen, the vials are sealed in the device, and the stoppers are secured with a flap flap. .

The vials are inspected visually for closure and external defects. Withered vials are discarded and destroyed.

Cetrorelix lophilisate 1 mg is a white, baked cake which is freeze-dried in a colorless manner. 2 ml vial, which is closed with a freeze-dried gray stopper and a yellow flipp-off bead cap.

Claims (6)

A lyophilizate having a peptide of 3 to 15 amino acids and optionally one or more scaffolding agents, characterized in that 1 unit by weight of the peptide is dissolved in 100 to 15 10,000 parts by weight of acetic acid are then taken up in water and the solution thus obtained is lyophilized. Lyophilisate according to claim 1, characterized in that altered is used as the scaffolding agent. Lyophilisate according to claim 1, characterized in that the peptide is cetrorelix. 4. Use of a lyophilizer, wrobed 1 or 2 do not treat female infertility. p. Use of the Ivofilisate obtained according to claim 1 1 or 2 to protect the gonads. Use of the lyophilizate obtained according to claim 1 aleco 2pr · drug production r.and treatment of female infertility. Use of the lyophilizate obtained according to claim 1 or 2 for the manufacture of a medicament for the protection of the gonads.