(19)
(11)EP 3 590 524 B1

(12)EUROPEAN PATENT SPECIFICATION

(45)Mention of the grant of the patent:
04.11.2020 Bulletin 2020/45

(21)Application number: 18181931.9

(22)Date of filing:  05.07.2018
(51)International Patent Classification (IPC): 
A61K 38/09(2006.01)
A61K 31/185(2006.01)
A61K 9/19(2006.01)
F26B 5/06(2006.01)

(54)

A METHOD OF RECONSTITUTING A TEVERELIX-TFA COMPOSITION

EINE METHODE ZUR REKONSTITUTION EINER TEVERELIX-TFA-ZUSAMMENSETZUNG

UNE MÉTHODE POUR RECONSTITUER UNE COMPOSITION TEVERELIX-TFA


(84)Designated Contracting States:
AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

(43)Date of publication of application:
08.01.2020 Bulletin 2020/02

(73)Proprietor: Antev Limited
London W1S 1DN (GB)

(72)Inventors:
  • LARSEN, Finn
    Hawick, TD9 8JS (GB)
  • BOUTIGNON, François
    63000 Clermont-Ferrand (FR)
  • POLAND, Guy
    Bristol, BS40 5SG (GB)

(74)Representative: Holme Patent A/S 
Valbygårdsvej 33
2500 Valby
2500 Valby (DK)


(56)References cited: : 
WO-A1-2008/071984
US-A1- 2003 044 463
  
      
    Note: Within nine months from the publication of the mention of the grant of the European patent, any person may give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall be filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has been paid. (Art. 99(1) European Patent Convention).


    Description


    [0001] The present invention relates to a method of reconstituting said composition.

    [0002] Teverelix is a synthetic gonadotropin-releasing hormone antagonists (GnRH antagonists) that compete with the endogenous neurohormone GnRH (otherwise known as luteinizing hormone releasing hormone, LHRH) for binding to its receptors in the anterior pituitary gland. By decreasing or blocking GnRH action, the GnRH antagonist suppress release from the anterior pituitary gland of follicle stimulating hormone (FSH) and luteinizing hormone (LH).

    [0003] Both FSH and LH are involved in normal reproductive function. In females, FSH stimulates the growth of immature Graafian follicles to maturation, whereas changes in LH levels control ovulation. In males, on the other hand, FSH plays an important role in spermatogenesis and LH stimulates production of testosterone in the testes.

    [0004] Accordingly, teverelix is suitable for treatment of hormone-dependent conditions such as benign prostatic hypertrophy, hormone-dependent prostate cancer, endometriosis and uterine myomas.

    [0005] Since teverelix (Ac-D-Nal-D-pClPhe-D-Pal-Ser-Tyr-D-Hci-Leu-Lys(iPr)-Pro-D-Ala-NH2) is a hydrophobic peptide it has a tendency of forming gels in the presence of many counter-ions. This problem has been solved in WO 2003/022243 which discloses that the formation of a gel may be prevented by contacting the teverelix peptide with a counter-ion, e.g. trifluoroacetate (TFA), at a molar ratio of at least 1:1.6 of peptide to counter-ion, thereby providing a fluid, milky microcrystalline aqueous suspension of the teverelix-salt (e.g. teverelix-TFA). According to WO 2003/022243, a ratio of teverelix to the counter-ion trifluoroacetate (TFA) of at least 1:1.6 is essential in order to ensure the desired microcrystalline suspension is obtained, otherwise a gel will be formed. However, the inventors of the present invention has found that the molar ratios disclosed in WO 2003/022243 will result in both undesirable gel-formation and in suspensions which are not homogenous. This is a problem, not only because such suspensions will be difficult to inject, but also because the bioavailability of the teverelix peptide is compromised since the gel interferes with the desired sustained action of said peptide.

    [0006] Furthermore, since teverelix is deamidated when placed in contact with water and acid, the teverelix-TFA compositions of WO 2003/022243 is not stable during storage e.g. at refrigeration and room temperatures and accordingly the teverelix composition has a relatively short shelf life under such conditions.

    [0007] Deamidation is caused by hydrolysis of the teverelix peptide. However since a reduction in the water content may contribute to an increased rate of deamidation, e.g. due to a lower pH-value in the composition, it would not be obvious to reduce the water content in the composition in order to increase the shelf life.

    [0008] Another problem with the teverelix-TFA compositions disclosed in WO 2003/022243 is that the inventors of the present invention has discovered that there are batch variations in the molar ratio of teverelix to the counter-ion TFA provided by the manufactures, even though the applied manufacturing conditions and processes are identical.

    [0009] Since the molar ratio is essential for obtaining the desired microcrystalline suspension, variations in said ratio may affect the bioavailability of teverelix in the pharmaceutical formulations. It is therefore essential that medical personal and other users can rely on the molar ratio of teverelix to trifluoroacetate provided by the manufactures.

    [0010] Accordingly there is a demand to both develop stable compositions having a long shelf life, and provide new methods of manufacturing teverelix-TFA in which a predefined molar ratio of peptide to counter-ion is obtained and which corresponds to the expectations.

    [0011] Thus, it is a first aspect to provide a teverelix-TFA composition having a prolonged shelf-life at refrigerating and room temperature.

    [0012] It is a second aspect to provide a method of ensuring that the desired molar ratio of peptide to counter-ion is obtained.

    [0013] It is a third aspect of the present invention to provide a method of adjusting the molar ratio of peptide to counter-ion in a solution.

    [0014] It is a fourth aspect to provide teverelix-TFA compositions that exhibit a long duration of action, with a lower injection volume and which further allow for less frequent administration.

    [0015] These and further aspects are achieved by providing a reconstitutable teverelix-TFA composition having a molar ratio of teverelix to trifluoroacetate which is below the molar ratio required for microcrystal formation.

    [0016] Since the reconstitutable composition has a low content of the counter-ion trifluoroacetate the pH value in the reconstitutable composition is maintained at a value which ensures that the deamidation of teverelix is reduced to an acceptable level, i.e. the possible deamidation which will take place is reduced to a level were it will not effect the stability of teverelix, whereby teverelix remains stable during storage e.g. at a temperature around 2-8°C.

    [0017] It is in this respect preferred that the molar ratio of teverelix to trifluoroacetate in the reconstitutable composition is between 1:1 and 1:1.85, preferably around or below 1:1.6, as this will provide a stable reconstitutable teverelix-TFA composition

    [0018] In a preferred embodiment the molar ratio of teverelix to trifluoroacetate (TFA) is above 1:1.3 and below 1:1.6, as the inventors of the present invention has found that molar ratios in this range provides an optimal stability of the reconstitutable teverelix-TFA composition.

    [0019] Reconstitutable compositions are well known in the pharmaceutical industry, where water or another diluent is added to the dry composition immediately prior to administration. Such conventional reconstitutable compositions are stored in a substantially dry state in order to maintain the stability of the composition. The inventors of the present invention has however found that if a small amount of water is present in the reconstitutable compositions, i.e. in an amount between 0.3% to 5% by weight, preferably around 1 to 2% by weight, based on the total weight of the reconstitutable teverelix-TFA composition, an improved teverelix-TFA composition is provided which is easier to handle, reconstitute, and accordingly use.

    [0020] Without being bound by theory, the water content may provide high electrostatic forces between particles of teverelix, which is of importance when the reconstitutable composition is handled e.g. when the composition is filled to a vial or a syringe chamber.

    [0021] In a preferred embodiment water is present in the reconstitutable teverelix-TFA composition in an amount between 1% by weight and 2% by weight, preferably 1.5% by weight, based on the total weight of the reconstitutable teverelix-TFA composition, as this will provide a reconstitutable teverelix-TFA composition that retains its chemical integrity and provides a stable composition.

    [0022] The stability provided enables a longer shelf-life at room temperature so that the reconstitutable teverelix-TFA composition may be stored e.g. after sterilization. The reconstitutable composition can be packaged and stored (e.g., in a syringe or vial) for later use.

    [0023] The reconstitutable composition may be prepared/manufactured in any suitable way, but in one embodiment teverelix and trifluoroacetate are contacted in an aqueous solution in order to provide an aqueous teverelix-TFA solution, which then is dried, e.g. freeze-dried or spray dried, in order to provide the reconstitutable composition. The teverelix and trifluoroacetate are in one embodiment contacted at a predefined molar ratio which is below the molar ratio required for microcrystal formation, preferably above 1:3 and below 1:1.6.

    [0024] In a preferred embodiment the drying step is ceased when the reconstitutable composition contains water in an amount between 0.3% to 5% by weight based on the total weight of the reconstitutable teverelix-TFA composition. Preferably the solution is dried such that water is present in the reconstitutable teverelix-TFA composition in an amount between 1% by weight and 2% by weight, e.g. 1.5% by weight. The amount of water in said composition may e.g. be evaluated at the end of the drying period using conventional measuring methods and equipment.

    [0025] As discussed earlier batch variations in the molar ratio of teverelix to trifluoroacetate may, for unknown reasons, arise during manufacturing, however it is preferred to know the exact molar ratio of teverelix to trifluoroacetate in a specific reconstitutable teverelix-TFA composition. Accordingly, the method of manufacturing the reconstitutable teverelix-TFA composition may further comprise the step of analysing the molar ratio in the solution prior to drying, such that the specific molar ratio is known. This will also provide the possibility of adjusting said molar ratio to a specific predefined molar ratio by adding trifluoroacetate before the solution is dried, thereby effectively counteracting any variation in the molar ratio that may exist. Thus, if the molar ratio is found to be e.g. 1:1.2 in the solution, said molar ratio may be adjusted to 1:1.4 by adding trifluoroacetate. The only requirement being that the resultant molar ratio before drying has to be below the molar ratio required for microcrystal formation.

    [0026] The molar ratio of the solution may be determined by first measuring the content of trifluoroacetate and teverelix in the sample using a conventional HPLC method, and then calculating the molar ratio using the following formula:



    [0027] The molar mass of trifluoroacetate and teverelix has been calculated to: MTFA = 114 g/mol and MTev = 1459 g/mol.

    [0028] After drying the thereby obtained reconstitutable teverelix-TFA composition may be stored and reconstituted with water or another suitable solution when an injectable pharmaceutical formulation is to be prepared.

    [0029] Providing a reconstitutable teverelix-TFA composition with a specific and predefined molar ratio will ensure that a fixed amount of trifluoroacetate can be added to said composition upon reconstitution thereby effectively and simply achieving an exact and desired molar ratio in order to obtain the fluid, milky microcrystalline aqueous suspension of the teverelix salt: teverelix-TFA, without formation of a gel. According to the present invention the molar ratio of teverelix to TFA after reconstitution is adjusted to at least 1:2.1 as the inventors have shown that a teverelix-TFA microcrystal suspension without formation of a gel thereby is obtained. When the ratio is above 1:2.2 the suspension is also homogeneous.

    [0030] Thus, the low molar ratio of teverelix to the counter-ion trifluoroacetate in the reconstitutable composition will, in addition to optimising the stability of the teverelix peptide, also ensure that it is possible to adjust the molar ratio of peptide to trifluoroacetate during the reconstitution process thereby ensuring that the optimal molar ratio of peptide to counter-ion may be obtained (depending on the intended use of the reconstituted composition) simply by adding the sufficient amount of trifluoroacetate during or after reconsitution.

    [0031] For instance, if during the manufacturing process it has been established that the reconstitutable teverelix-TFA composition has a molar ratio of 1:1.4, and if a molar ratio of 1:2.2 is desired in the final aqueous teverelix-TFA formulation, then 0.8 mol TFA per mol teverelix present in the composition has to be added during the reconstitution process according to the invention. Said amount can easily be calculated by a person skilled in the art.

    [0032] A person skilled in the art will further understand that as long as the molar ratio of teverelix to trifluoroacetate is below the molar ratio required for microcrystal formation, it is of no relevance if said molar ratio is e.g. 1:1.4 or 1:1.36 as long as said ratio is known or can be calculated, in order to ensure that a person skilled in the art can calculate the correct amount of TFA to be used during the reconstitution process.

    [0033] The trifluoroacetate content may be added/adjusted after the composition is reconstituted, but in a preferred embodiment the trifluoroacetate is part of the aqueous reconstitution solution, as this will provide a fast and effective way of reconstituting the composition. If desired, the aqueous reconstitution solution may contain an isotonic agent, such as mannitol and/or a pharmaceutically acceptable excipient.

    [0034] A person skilled in the art will based on the present application understand that instead of drying the teverelix-TFA solution in order to obtain a reconstitutable teverelix-TFA composition, the molar ratio can be adjusted directly in the solution to a molar ratio of teverelix to trifluoroacetate which is above the molar ratio required for microcrystal formation, i.e. above a ratio of 1:2.1, preferably at least 1:2.2 and even more preferred at least 1:2.4 after having evaluated the exact molar ratio, by adding a sufficient amount of trifluoroacetate. This will provide an aqueous pharmaceutical formulation that may be used directly, i.e. the formulation is ready-to-use.

    [0035] The inventors of the present invention have furthermore found that when the molar ratio of teverelix to trifluoroacetate is at least 1:2.1, preferably at least 1:2.2, in the final aqueous pharmaceutical formulation, the formulation will comprise both soluble and insoluble teverelix, thereby providing a unique bioavailablity of teverelix.

    [0036] Without being bound by theory, the soluble teverelix is in the form of an aqueous solution and in some situations, a gel. The presence of a gel will inhibit any freely aqueous teverelix and therefore prevent, or at least reduce, immediate release. The insoluble teverelix is in the form of microcrystals. Said microcrystals will prevent gel formation, therefore "unlocking" the aqueous teverelix. Over time the TFA in the composition will be absorbed by the body, lowering the ratio, so the microcrystals subsequently turn in to gel, which forms the slow release depot. Thus, the non-gel-soluble teverelix is immediately available, providing an almost immediate onset of action, and the gel-soluble and insoluble teverelix (microcrystals) will assist in providing a sustained release of teverelix.

    [0037] Accordingly, using the teverelix-TFA composition described herein, it is possible to adjust the release profile of teverelix simply by adjusting the amount of trifluoroacetate added to the reconstitutable composition and thereby change the ratio of insoluble to soluble teverelix in the injected composition.

    [0038] However in a simple and preferred embodiment, the reconstitutable teverelix-TFA composition is provided as a unit dosage having a predefined molar ratio of teverelix to the counter-ion TFA which is below the molar ratio required for microcrystal formation, preferably between 1:1 and 1:1.85 and even more preferred above 1:1.3 and below 1:1.6, and wherein said unit dosage is reconstituted just prior to use by adding a fixed amount of aqueous reconstitution solution containing a sufficient amount of trifluoroacetate so that a predetermined molar ratio of teverelix to trifluoroacetate is achieved in the provided pharmaceutical formulation. Said predetermined molar ratio is preferably at least 1:2.1, preferably at least 1:2.2 and even more preferred about 1:2.4, thereby ensuring that the provided reconstituted suspension contains substantially no gel, or at least so small concentrations of gel that the reconstituted suspension can be used for injections. Accordingly, another aspect of the application features a package e.g. a syringe or vial, filled with a unit dosage of the reconstitutable teverelix-TFA composition. Within the context of the present application the term "unit dosage" is the amount of an active ingredient (teverelix) administered to a patient in a single dosage. Said unit dosages is e.g. placed in a suitable syringe in order to provide an easy administration.

    [0039] In a preferred embodiment the invention also relates to a kit, comprising a first package filled with a unit dosage of the teverelix-TFA composition, and a second package filled with a reconstitution solution comprising a sufficient amount of TFA for obtaining the desired molar ratio of at least 1:2.1, preferably at least 1:2.2 and even more preferred about or above 1:2.4. Said first package may e.g. be a syringe and the second package may be physically connected to said syringe in order to ensure that the correct molar ratio of teverelix to TFA is obtained. As one example of a first and second package which is physically connected to each other can be mentioned a conventional dual chamber syringe for lyophilised products. Such dual chamber syringe are well known in the art.

    [0040] In one embodiment said kit is arranged for providing a final teverelix-TFA formulation having a molar ratio of teverelix to counter-ion of 1:2.4. Preferably the concentration of teverelix is between 30 mg/ml and 100 mg/ml, and even more preferred between 45 mg/ml and 90 mg/ml, e.g. about 75 mg/ml. The concentration of teverelix may in some situations be higher than about 100 mg/ml. The volume may be between 0.4 ml and 1.5 ml, e.g. about 1.2 ml. Injection given subcutaneous and/or intramuscularly at this concentration and volume, has proven to only provide a mild injection site reaction.

    [0041] The compositions and formulations provided in the present application is inexpensive to manufacture, and due to the ease of use it provides a very simple dosage regime.

    [0042] Modifications and combinations of the above principles and combinations are foreseen within the scope of the present invention.

    Examples:



    [0043] In order to establish the influence of the molar ratio of teverelix to the counter-ion trifluoroacetate a number of tests were performed.

    Example 1: Preparation of teverelix-TFA compositions with different molar ratio



    [0044] A custom-manufactured batch (Batch A) of teverelix with low TFA content was obtained. The characteristics of Batch A are shown in table 1.
    Table 1
    Purity 99.3 %
    Teverelix content 85.56 weight-%
    TFA content 10.9 weight-%
    Acetate content 0.3 weight-%
    Water content 4.3 weight-%


    [0045] If a composition containing 75 mg teverelix is desired, composition A, then 88.28 mg of batch A has to be used, calculated as follows:



    [0046] The molar ratio of teverelix to TFA in composition A can then be calculated.

    [0047] The TFA content in 88.28 mg of batch A can be calculated to: 88.28 mg x 10.9/100 (TFA content in %) = 9.62 mg

    [0048] Since the molar mass of TFA, MTFA, is 114 g/mol, and the molar mass of teverelix, MTEV, is 1459 g/mol, the molar concentration of TFA in the 75mg teverelix composition can be calculated to 0.084 mmol and the molar concentration of teverelix to 0.051 mmol. Thus, the molar ratio of teverelix to TFA in the reconstitutable composition A is 1:1.64.

    [0049] In order to prepare a number of different aqueous teverelix-TFA compositions with different molar ratios, twenty-one samples containing 44.14 mg + 5% (41.93 to 46.35 mg) of composition A were accurately weighed in 2 ml glass tubes having a cap through which a reconstitution solution could be added by means of a micropipette.

    [0050] Seven TFA reconstitution solutions containing TFA in 5% mannitol were prepared using a TFA composition obtained from Acros Organics, Geel, Belgium. Said TFA composition were 99 % pure and had a density of 1.535 g/ml. The respective reconstitution solutions are shown in Table 2.
    Table 2
    SolutionABCDEFG
    TFA mol/L 0 0.01 0.023 0.036 0.049 0.062 0.075


    [0051] The respective aqueous teverelix-TFA compositions were prepared by adding 0.5 ml of each of the above reconstitution solutions though the cap of the twenty-one glass tubes containing 44.14 mg + 5% (41.93 to 46.35 mg) of composition A using a micropipette, i.e. three aqueous teverelix-TFA compositions having the same molar ratio were prepared. The mixtures were stirred using a vortex for 1 minute, and the solutions were observed visually for 10 minutes in order to establish if the desired fluid, milky microcrystalline homogeneous aqueous suspension of the teverelix-TFA, were obtained, or if a gel was formed instead. The results are summarised in Table 3 below:
    Table 3
    TubesMolar ratioFormation of gelMicrocrystalline formationFormation of milky suspensionhomogeneous suspension
    A1, A2, A3 1:1.64 yes no no -
    B1, B2, B3 1:1.85 yes no no -
    C1, C2, C3 1:2.1 no yes yes no
    D1, D2, D3 1:2.36 no yes yes yes
    E1, E2, E3 1:2.61 no yes yes yes
    F1, F2, F3 1:2.86 no yes yes yes
    G1, G2, G3 1:3.12 no yes yes yes


    [0052] The microcrystalline content of the aqueous teverelix-TFA compositions in the No. 1 test tubes were further observed under a polarized light microscope supplied by Realux, France. The results for the respective molar ratio are shown in fig. 1a - fig. 1g. From these observations it is clear that microcrystalline formation is not observed for the molar ratios of 1:1.85 and below, thus the molar ratio of teverelix to the counter-ion TFA has to be above at least 1:1.85 in order for the desired microcrystalline formation to be initiated.

    [0053] It is accordingly preferred that the molar ratio of teverelix to trifluoroacetate (TFA) in the reconstitutable composition is below 1:1.85 and preferable below 1:1.6.

    [0054] Furthermore, as is evident from table 3, a homogeneous suspension of teverelix-TFA was not obtained until the molar ration was above 1:2.1, thus it is accordingly preferred that the molar ratio in the reconstituted aqueous teverelix-TFA suspension is above 1:2.1 and preferably even higher such as at least 1:2.2, and even more preferred at least 1:2.4.

    Example 2: Content of soluble teverelix and insoluble teverelix in relation to the molar ratio.



    [0055] In order to determine the content of soluble teverelix in relation to insoluble teverelix in the respective test tubes, the No. 2 and No. 3 test tubes for each molar ratio were centrifuged at 10,000 rpm for 10 to 20 minutes, and the concentration of teverelix in the supernatant and pellet were measured using a HPLC analysis.

    [0056] The chromatographic conditions for the HPLC analysis is shown in table 4.
    Table 4
    Column Type (Aptys N°) Lichrospher 100 RP18 (N°128)
      Particles size 5 µm
      Diameter 4 mm
      Length 125 mm
    Pre-Column Type Lichrocart 100 RP18
      Particles size 5 µm
      Diameter 4 mm
      Length 4 mm
    Mobile Phase Acetonitrile/Water/TFA (35:65:0.1 V/V/V)
    Injector cleaning Acetonitrile/Water (50:50 V/V)
    Flow 1.0 mL/min
    Pressure Approx. 65 bars
    Oven Temperature 30°C
    Wavelength 210 nm
    Injection volume 10 µL
    Injector temperature 20°C
    Retention time of Teverelix Approx. 5.6 min
    Run time 10 min


    [0057] Two 100% standards were prepared by weighing 59.9 mg teverelix acetate (batch 080113) in a volumetric flask and completing the volume to 100 ml with water: acetonitrile 65:35 v/v. 10 ml of this solution were completed to 50 ml with the same solvent, providing a concentration of 0.1 mg/ml teverelix peptide.

    [0058] A 1% standard solution was prepared by diluting 2 ml of the 100% standard to 200 ml with the same solvent providing a concentration of 0.001 mg/ml teverelix peptide.

    [0059] Internal standardization was carried out using the two 100% standards. The 1% standard was used to check the linearity of the response. Recovery with the 100% standard must be in the interval 95%-105%.

    [0060] The pellet obtained after centrifugation was solubilised in water:acetonitrile 65:35 v/v, and the volume was completed to 100 mL with the same solvent. This solution was diluted by 5 (10 mL in 50 mL) and HPLC was performed.

    [0061] The supernatant was transferred to a volumetric flask and the volume was completed to 100mL with the same solvent, i.e. water:acetonitrile 65:35 v/v. This solution was diluted by 5 (10 mL in 50 mL) and HPLC was performed. The results of the HPLC analysis is shown in table 5.
    table 5
    Test tubeMolar ratioSupernatant - Teverelix concentration (mg/ml)Pellet - Teverelix concentration (mg/ml)
    A2 1:1.64 52.0 N/A
    A3 1:1.64 58.5 N/A
    B2 1:1.85 57.2 N/A
    B3 1:1.85 60.3 N/A
    C2 1:2.1 25.9 26.9
    C3 1:2.1 26.1 25.5
    D2 1:2.36 9.4 39.3
    D3 1:2.36 8.3 44.9
    E2 1:2.61 5.4 50.8
    E3 1:2.61 7.2 51.6
    F2 1:2.86 3.7 56.2
    G3 1:2.86 3.6 58.4
    G2 1:3.12 1.5 53.6
    G3 1:3.12 1.2 58.4


    [0062] The average concentrations of each molar ratio were calculated, see table 6, and the results are depicted in fig. 2 and 3.
    Table 6
    Test tubeMolar ratioSupernatant - Average Teverelix concentration (mg/ml)Pellet - Average Teverelix concentration (mg/ml)Total (pellet + supernatant) Teverelix concentration (mg/ml)
    A 1:1.64 55.3 N/A 55.3
    B 1:1.85 58.8 N/A 58.8
    C 1:2.1 26.0 26.2 52.2
    D 1:2.36 8.9 42.1 51.0
    E 1:2.61 6.3 51.2 57.5
    F 1:2.86 3.7 57.3 61.0
    G 1:3.12 1.4 56.0 57.4


    [0063] As is evident from table 5, and 6, and fig. 2 and 3, the degree of insoluble teverelix increases when the amount of trifluoroacetate increases in relation to teverelix, thus at a molar ratio of 1:2.1, about 50 % of the pharmaceutical formulation consist of insoluble teverelix, whereas the amount of insoluble teverelix is about 82 % at a molar ratio of 1:2.36 (∼1:2.4) in the pharmaceutical formulation.

    Example 3: Plasma concentration in relation to the molar ratio.



    [0064] In order to evaluate the relevance of the molar ratio on the plasma concentration of teverelix, five glass vials containing different molar ratios were prepared as discussed in example 1, and the test tubes comprising the aqueous teverelix-TFA compositions shown in table 7 were provided:
    Table 7
    TubeIIIIIIIVV
    Molar ratio 1.64 2.1 2.36 2.61 2.86


    [0065] Five rats were tested with each molar ratio. Each rat was injected with 60µl of the respective solutions using a 25mm 21G luer 6% regular bevel needle (obtainable from Terumo, Leuven, Belgium) and 100µl luer slip syringe (obtainable from Hamilton Company, Reno, USA). Plasma concentrations were measured prior to administration, then at 1h, 6h, 24h, 48h, 7 days, 10 days, 14 days, 21 days and 28 days following administration.

    [0066] The peak plasma concentrations, Cmax, of teverelix after injection to each individual rat are shown in table 8, and depicted in fig. 4.
    Table 8
    Test tubeMolar ratioCmaxCmaxCmaxCmaxCmaxCmax meanCmax median
    I 1:1.64 57.6 58.8 35.4 32.5 25 41.86 35.4
    II 1:2.1 96 82.6 57.4 50.1 n.a. 76.525 70
    III 1:2.36 67.6 50 67.9 64.2 88.6 67.66 67.6
    IV 1:2.61 78.8 48.6 85.5 77.5 55.3 69.14 77.5
    V 1:2.86 111 99.7 94.9 91.9 84.8 96.46 94.9


    [0067] As is clear from these results the Teverelix Cmax increases until a molar ratio of 1:2.1 after which the plasma concentration is substantially stable.

    [0068] The plasma concentration over a four week period, was also measured by taking blood samples at regular intervals.

    [0069] The mean plasma levels in a four weeks period is shown in fig. 5, and it is clear that the release profile of teverelix is dependent on the molar ratio. For instance a higher plasma concentration of teverelix is shown with the suspension having a molar ratio of 1:2.1. Thus, it is possible to adjust the release profile of teverelix simply by adjusting the amount of trifluoroacetate added to the reconstitutable composition, thereby changing the molar ratio of teverelix to trifluoroacetate in the pharmaceutical formulation. Clinically this offers the potential of optimising the therapy to the requirements of individual groups of patients e.g. relating to different indications, age and/or gender. One patient group may need an immediate onset of action, requiring a high concentration of soluble teverelix, whereas another group may require a sustained release of teverelix, requiring a low concentration of soluble teverelix. In a similar manner, different pharmaceutical formulations having different molar ratios may be administered at different stages of a patients treatment. Furthermore, the possibility of adjusting the molar ratio to specific needs of different patient groups, will increase patient acceptance and compliance of therapy.


    Claims

    1. A method of reconstituting a reconstitutable teverelix-TFA composition having a molar ratio of teverelix (Ac-D-Nal-D-pClPhe-D-Pal-Ser-Tyr-D-Hci-Leu-Lys(iPr)-Pro-D-Ala-NH2) to trifluoroacetate below the molar ratio required for microcrystal formation, wherein said molar ratio of teverelix to trifluoroacetate (TFA) is between 1:1 and 1:1.85, said method comprises adding an aqueous reconstitution solution to the reconstitutable teverelix-TFA composition and adjusting the molar ratio of teverelix to trifluoroacetate to at least 1:2.1 by adding trifluoroacetate.
     
    2. The method according to claim 1, wherein the aqueous reconstitution solution comprises trifluoroacetate in an amount sufficient to provide a molar ratio of teverelix to trifluoroacetate of at least 1:2.1, preferably at least 1:2.2 and even more preferred to at least 1:2.4.
     
    3. The method according to claim 1 or 2, wherein the molar ratio of teverelix to trifluoroacetate (TFA) in the reconstitutable teverelix-TFA composition is above 1:1.3 and below 1:1.6.
     
    4. The method according to claim 1, 2 or 3, wherein water is present in the reconstitutable teverelix-TFA composition in an amount between 0.3% to 5% by weight based on the total weight of the reconstitutable teverelix-TFA composition.
     
    5. The method according to any of the preceding claims, wherein water is present in the reconstitutable teverelix-TFA composition in an amount between 1% by weight and 2% by weight, preferably 1.5% by weight, based on the total weight of the reconstitutable teverelix-TFA composition.
     
    6. A kit comprising a first package filled with the reconstitutable teverelix-TFA composition having a molar ratio of teverelix (Ac-D-Nal-D-pClPhe-D-Pal-Ser-Tyr-D-Hci-Leu-Lys(iPr)-Pro-D-Ala-NH2) to trifluoroacetate below the molar ratio required for microcrystal formation, wherein said molar ratio of teverelix to trifluoroacetate (TFA) is between 1:1 and 1:1.85 , and a second package filled with a aqueous reconstitution solution comprises trifluoroacetate in an amount sufficient to provide a molar ratio of teverelix to trifluoroacetate of at least 1:2.1 after reconstitution.
     
    7. The kit according to claim 6, wherein the first package comprising a unit dosage of the reconstitutable teverelix-TFA composition.
     
    8. The kit according to claim 6 or 7, wherein the first package is a syringe suitable for providing a subcutaneous and/or intramuscularly injection.
     


    Ansprüche

    1. Verfahren zum Rekonstituieren einer rekonstituierbaren Teverelix-TFA-Zusammensetzung mit einem Molverhältnis von Teverelix (Ac-D-Nal-D-pClPhe-D-Pal-Ser-Tyr-D-Hci-Leu-Lys(iPr)-Pro-D-Ala-NH2) zu Trifluoracetat unterhalb des für die Mikrokristallbildung erforderlichen Molverhältnisses, wobei das Molverhältnis von Teverelix zu Trifluoracetat (TFA) zwischen 1:1 und 1:1.85 liegt, das Verfahren das Zugeben einer wässrigen Rekonstitutionslösung zu der rekonstituierbaren Teverelix-TFA-Zusammensetzung und das Einstellen des Molverhältnisses von Teverelix zu Trifluoracetat auf mindestens 1:2.1 durch Zugabe von Trifluoracetat umfassend.
     
    2. Verfahren nach Anspruch 1, wobei die wässrige Rekonstitutionslösung Trifluoracetat in einer Menge umfasst, die ausreicht, um ein Molverhältnis von Teverelix zu Trifluoracetat von mindestens 1:2.1, vorzugsweise mindestens 1:2.2 und noch bevorzugter von mindestens 1:2.4, bereitzustellen.
     
    3. Verfahren nach Anspruch 1 oder 2, wobei das Molverhältnis von Teverelix zu Trifluoracetat (TFA) in der rekonstituierbaren Teverelix-TFA-Zusammensetzung über 1:1.3 und unter 1:1.6 liegt.
     
    4. Verfahren nach Anspruch 1, 2 oder 3, wobei Wasser in der rekonstituierbaren Teverelix-TFA-Zusammensetzung in einer Menge zwischen 0.3 und 5 Gew.-%, bezogen auf das Gesamtgewicht der rekonstituierbaren Teverelix-TFA-Zusammensetzung, vorhanden ist.
     
    5. Verfahren nach einem der vorstehenden Ansprüche, wobei Wasser in der rekonstituierbaren Teverelix-TFA-Zusammensetzung in einer Menge zwischen 1 Gew.-% und 2 Gew.-%, vorzugsweise 1.5 Gew.-%, bezogen auf das Gesamtgewicht der rekonstituierbaren Teverelix-TFA-Zusammensetzung, vorhanden ist.
     
    6. Kit, einen ersten Behälter umfassend, der mit der rekonstituierbaren Teverelix-TFA-Zusammensetzung gefüllt ist, die ein Molverhältnis von Teverelix (Ac-D-Nal-D-pClPhe-D-Pal-Ser-Tyr-D-Hci-Leu-Lys(iPr)-Pro-D-Ala-NH2) zu Trifluoracetat unterhalb des für die Mikrokristallbildung erforderlichen Molverhältnisses aufweist, wobei das Molverhältnis von Teverelix zu Trifluoracetat (TFA) zwischen 1:1 und 1:1.85 liegt, und einen zweiten Behälter, der mit einer wässrigen Rekonstitutionslösung gefüllt ist, die Trifluoracetat in einer Menge umfasst, die ausreicht, um nach der Rekonstitution ein Molverhältnis von Teverelix zu Trifluoracetat von mindestens 1:2.1 bereitzustellen.
     
    7. Kit nach Anspruch 6, wobei der erste Behälter eine Dosierungseinheit der rekonstituierbaren Teverelix-TFA-Zusammensetzung umfasst.
     
    8. Kit nach Anspruch 6 oder 7, wobei der erste Behälter eine Spritze ist, die für das Bereitstellen einer subkutanen und/oder intramuskulären Injektion geeignet ist.
     


    Revendications

    1. Procédé de reconstitution d'une composition de teverelix-TFA apte à être reconstituée qui possède un rapport molaire de la teverelix (Ac-D-Nal-D-pCIPhe-D-Pal-Ser-Tyr-D-Hci-Leu-Lys(iPr)-Pro-D-Ala-NH2) au trifluoroacétate inférieur au rapport molaire requis pour la formation de microcristaux ; dans lequel ledit rapport molaire de la teverelix au trifluoroacétate (TFA) se situe entre 1:1 et 1:1,85; ledit procédé comprenant le fait d'ajouter une solution aqueuse de reconstitution à la composition de teverelix-TFA apte à être reconstituée et le fait d'ajuster le rapport molaire de la teverelix au trifluoroacétate jusqu'à une valeur qui s'élève à au moins 1:2,1 par l'intermédiaire d'une addition de trifluoroacétate.
     
    2. Procédé selon la revendication 1, dans lequel la solution aqueuse de reconstitution comprend du trifluoroacétate en une quantité qui est suffisante pour procurer un rapport molaire de la teverelix au trifluoroacétate qui s'élève à au moins 1:2,1, de préférence à au moins 1:2,2, et de manière encore plus préférée à au moins 1:2,4.
     
    3. Procédé selon la revendication 1 ou 2, dans lequel le rapport molaire de la teverelix au trifluoroacétate (TFA) dans la composition de teverelix-TFA apte à être reconstituée est supérieur à 1:1,3 et est inférieur à 1:1,6.
     
    4. Procédé selon la revendication 1, 2 ou 3, dans lequel de l'eau est présente dans la composition de teverelix-TFA apte à être reconstituée, en une quantité qui représente entre 0,3 % et 5 % en poids en se basant sur le poids total de la composition de teverelix-TFA apte à être reconstituée.
     
    5. Procédé selon l'une quelconque des revendications précédentes, dans lequel de l'eau est présente dans la composition de teverelix-TFA apte à être reconstituée, en une quantité qui représente entre 1 % en poids et 2 % en poids, de préférence qui représente 1,5 % en poids, en se basant sur le poids total de la composition de teverelix-TFA apte à être reconstituée.
     
    6. Nécessaire comprenant un premier lot qui est rempli avec la composition de teverelix-TFA apte à être reconstituée qui possède un rapport molaire de la teverelix (Ac-D-Nal-D-pClPhe-D-Pal-Ser-Tyr-D-Hci-Leu-Lys(iPr)-Pro-D-Ala-NH2) au trifluoroacétate inférieur au rapport molaire requis pour la formation de microcristaux ; dans lequel ledit rapport molaire de la teverelix au trifluoroacétate (TFA) se situe entre 1:1 et 1:1,85 ; et un second lot rempli avec une une solution aqueuse de reconstitution qui comprend du trifluoroacétate en une quantité suffisante pour obtenir un rapport molaire de la teverelix au trifluoroacétate jusqu'à une valeur qui s'élève à au moins 1:2,1 après la reconstitution.
     
    7. Nécessaire selon la revendication 6, dans lequel le premier lot comprend une unité posologique de la composition de teverelix-TFA apte à être reconstituée.
     
    8. Nécessaire selon la revendication 6 ou 7, dans lequel le premier lot représente une seringue qui est appropriée pour procurer une injection par voie sous-cutanée et/ou par voie intramusculaire.
     




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    Cited references

    REFERENCES CITED IN THE DESCRIPTION



    This list of references cited by the applicant is for the reader's convenience only. It does not form part of the European patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be excluded and the EPO disclaims all liability in this regard.

    Patent documents cited in the description