WO2019180072A1

WO2019180072A1 - Parenteral pharmaceutical composition comprising neladenoson bialanate

Info

Publication number: WO2019180072A1
Application number: PCT/EP2019/056943
Authority: WO
Inventors: Matthias Christian ERBER; Johanna Anlahr; Janine NICOLAI; Michael Pfeffer
Original assignee: Bayer Pharma Aktiengesellschaft
Priority date: 2018-03-22
Filing date: 2019-03-20
Publication date: 2019-09-26
Also published as: AR117406A1

Abstract

The present invention relates to parenteral pharmaceutical compositions comprising 2-{4-[2-({[2-(4-chlorophenyl)-1,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6-(pyrrolidin-1-yl)-pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate, also known as Neladenoson bialanate, in form of one of its salts, a process for its preparation, its use as medicament, as well as its use for prevention and/or treatment of cardiovascular diseases, e.g. the cardioprotection during and/or after an ischemia-induced cardiac event, e.g. after non-cardiac medium and major risk surgery, reperfusion injury in STEMI patients, and treatment of patients acutely hospitalized with worsening heart failure.

Description

PARENTERAL PHARMACEUTICAL COMPOSITION COMPRISING NELADENOSON BIALANATE

The present invention relates to parenteral pharmaceutical compositions comprising 2-{4- [2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6-(pyrrolidin-1 -yl)- pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate, also known as Neladenoson bialanate, in form of one of its salts, a process for its preparation, its use as medicament, as well as its use for prevention and/or treatment of cardiovascular diseases, e.g. the cardioprotection during and/or after an ischemia-induced cardiac event, e.g. after non-cardiac medium and major risk surgery, reperfusion injury in STEMI patients, and treatment of patients acutely hospitalized with worsening heart failure.

Neladenoson bialanate is the compound 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]- methyl}sulfanyl)-3,5-dicyano-6-(pyrrolidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-ala- ninate of the formula (I)

Neladenoson bialanate may be applied in free form or in form of a salt. Preferably the hy- drochloride (II), in particular the mono-hydrochloride (lla) is used.

Neladenoson is the compound 2-[[2-(4-chlorophenyl)thiazol-4-yl]methylsulfanyl]-4-[4-(2- hydroxyethoxy)phenyl]-6-pyrrolidin-1 -yl-pyridine-3,5-dicarbonitril of the formula (III)

Neladenoson, the bialanate as well as the hydrochloride and the mono-hydrochloride of the bialanate are known (see WO 2010/086101 and WO 2016/18871 1 ).

Neladenoson bialanate (hydrochloride) is the prodrug of the partial adenosine A1 receptor agonist Neladenoson and is currently being evaluated in clinical trials for the treatment of heart failure (of. ChemMedChem 2017, 12, 728-737).

Since poorly soluble drugs in their crystalline form often show poor bioavailability a pro drug approach is sometimes applied to improve both solubility and bioavailability like in the present case. While Neladenoson (III) is almost insoluble in aqueous media, the solu bility of the bialanate prodrug in such media is satisfactory. However, the stability of the prodrug leaves a lot to be desired, because the prodrug degrades of time forming Neladenoson (III).

Thus, to be suitable for parenteral, in particular intravenous, administration the stability of the active ingredient (here of the prodrug) is another key factor for success. In addition in creasing the solubility of Neladenoson, when formed by degradation, is a further challenge. Cyclodextrins (CD) are widely recognized pharmaceutical excipients used in numerous commercial products (see, e.g. WO 98/55148, US 6,407,079, EP-A 2 389 923, and Na ture Rev Drug Discovery 2004, 3, 1023-1035). They are inter alia described to be solubil izers for poorly soluble drugs (see e.g. Adv. Drug Deliv. Rev. 2007, 59, 645-666). Cy clodextrins are able to stabilize labile drugs in aqueous solution (see e.g. Int. J. Pharm. 2007, 329, 1 -1 1 ).

The addition of polyvinylpyrrolidone (PVP) to solutions of various poorly water soluble com pounds containing hydroxypropyl-3-cyclodextrin (HRbOϋ) is reported to either increase or decrease the solubility of the compound depending on the individual compound (see e.g. AAPS PharmSciTech 2005, 6(2) Article 43). PVP is also reported to increase solubility of poorly soluble drugs (without cyclodextrin addition) (see e.g. Int. J. Pharm. 1994, 110, 169- 177, ibid. 1998, 163, 1 15-121 ). This effect is reported to be concentration dependent, i.e. a higher PVP concentration leads to a higher solubility. However, this effect seems not al ways to be present (e.g. AAPS PharmSciTech 2009, 10(4), 1444-1451 ).

In general, degradation of a given active pharmaceutical ingredient can take place due to chemical instability, resulting in a new chemical entity. For example, chemical instability can result from hydrolysis and/or oxidation. Degradation of active pharmaceutical ingredi- ents is an undesired effect for pharmaceutical applications. The efficacy or availability of a drug can change dramatically.

An active pharmaceutical ingredient may also be physically unstable. Physical instability of a solution can result from crystallization and/or precipitation, when the solubility lim- it/solubility equilibrium of the active pharmaceutical ingredient is exceeded in a solution, which may result in particle formation. Particle formation in pharmaceuticals for parenteral application, in particular, makes the formulation harmful for clinical use.

Hence, bulk solutions used for further processing must show a sufficient physical and chemical stability (“holding time”) to ensure the integrity of the formulation. Ready-to-use solutions must also show a sufficient physical and chemical stability (“in-use stability”) to avoid harm when finally administered to the patient.

As is known in the field of pharmaceutical formulation, lyophilisation, also known as freeze-drying, is a method of processing a liquid product into a dry solid product. In gen eral, lyophilisation is defined as a stabilizing process in which the product is frozen foi lowed by elimination of the water content by sublimation. Lyophilisation methods are well known in the art (e.g. see Int. J. Pharm. 2000, 203, 1 -60).

The resulting lyophilised product should have an acceptable cake structure and sufficient stability (“shelf-life”), short rehydration/reconstitution time, and sufficient in-use- stability at the required temperature. One of the main disadvantages of lyophilized products is the fact that the stability of a drug depends on its physical state as well on the physical state of all components.

Prior to lyophilisation, a given active pharmaceutical ingredient, together with any excipi ents such as bulking agents, pH-adjusting agents, cryo protectors which may be present, for example, must be dissolved in a suitable solvent to form a solution (a“bulk solution”). After freeze-drying, the resulting dry solid product and, with that, the active ingredient, must again be dissolved in a suitable diluent or solvent to form a solution (a“reconstituted solution”). This reconstituted solution can then be administered to the patient directly or after further dilution.

A low filling volume of the vials is required in order to facilitate an efficient lyohilisation process. Therefore, lyophilisation is suitable only for active ingredients which allow for a high aqueous solubility or a low therapeutic dose in order to reduce the number of vials per administration. Thus, in case of high therapeutic doses the required number of vials per administration depends on the solubility of the drug substance.

Thus, the problem underlying this invention is the provision of a Neladenoson bialanate composition, wherein the Neladenoson bialanate is sufficiently stable and wherein possi- ble trace amounts of Neladenoson are sufficiently soluble, that the resulting composition is suitable for parenteral administration.

The invention thus provides a composition comprising

(a) 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6-(pyrro- lidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate (Neladenoson bialanate), (b) a chemically modified cyclodextrin,

(c) a polyvinylpyrrolidone polymer, and

(d) water.

The composition according to the invention is suitable for parenteral administration. Par enteral administration may take place by circumventing a bioabsorption step (for example intravenously, intraarterially, intracardially, intraspinally or intralumbarly), or with bioab sorption (for example intramuscularly, subcutaneously, intracutaneously, percutaneously or intraperitoneally). Intravenous, intraarterial and subcutaneous administrations are pre ferred, intravenous administration is particularly preferred. Administration forms suitable for parenteral administration are inter alia preparations for injection or infusion in the form of solutions, suspensions, emulsions, in liquid form, or as lyophilizates or sterile powders, which are reconstituted before administration.

Neladenoson bialanate may be applied in free form or in form of a salt. Preferably the hy drochloride (II), in particular the mono-hydrochloride (I la) is used. The mono-hydro- chloride contains the hydrochloride in an exact defined stoichiometric amount.

The Neladenoson bialanate is present in the compositions according to the invention in a concentration range of 0.5 to 50 mg/ml_ or 0.5 to 40 mg/ml_ or 1 to 40 mg/ml_ or 1 to 25 mg/ml_ or 1 to 5 mg/ml_, preferably in the range of 1 .5 to 3.5 mg/ml_, further preferred in the range of 2.25 to 3 mg/ml_, and further preferred with a concentration of 2.5 mg/ml_ [values calculated based on the compound of formula (I)].

The compounds according to formula (I), (II), (I la) and (III) act as partial adenosine A1 re ceptor agonists and may be used for the prevention and/or treatment of cardiovascular disorders like e.g. coronary heart disease, ischemic injury during acute coronary syn- drome, angina pectoris, heart failure, worsening chronic heart failure, heart failure with reduced ejection fraction (HFrEF), heart failure with preserved ejection fraction (HFpEF), myocardial infarction and atrial fibrillation

Cyclodextrins (CD) are cyclic oligosaccharides derived from starch containing six (aCD), seven (3CD), eight (yCD), nine (6CD), ten (sCD) or more (a-1 ,4)-linked a-D-glucopyran- ose units. Due to the chair conformation of the glucopyranose units, the CDs take the shape of a truncated cone or torus rather than a perfect cylinder. Several chemical modi fications of CDs are also of pharmaceutical interest, inter alia the hydroxypropyl deriva tives of 3CD and yCD (i.e. FIP3CD and FiPyCD), the hydroxyethyl derivative of 3CD (HE3CD), the methylated b-cyclodextrin (M3CD), the dimethyl-3-cyclodextrin (DM3CD), the randomly methylated 3CD (RM3CD), the permethylated b-cyclodextrin (PIV^CD), the sulfobutylether of bCD and yCD (i.e. SBEbCD and SBEyCD), and the so-called branched CDs such as maltosyl^CD (G2bCD) and glucosyl^-cyclodextrin (G^CD).

Within this invention the chemically modified CDs are applied. Preferably HIRbΰϋ and SBEbCD, particularly preferably F^CD, are used.

The amount of CD employed in the composition according to the invention may vary in a broad range. The composition according to the invention comprises preferably 10 to 75 % (w/v) of the chemically modified cyclodextrin, more preferably 20 to 50 % (w/v), even more preferably 30 to 40 % (w/v), particularly preferably 40 % (w/v). Such mass concen- trations relate to amounts of 10 to 750 mg/ml_, 20 to 50 mg/ml_, 30 to 40 mg/ml_ and 40 mg/ml_, respectively.

The polyvinylpyrrolidone (PVP) polymer is also known as povidone, povidonum, poly- vidone and poly(1 -vinyl-2-pyrrolidone) and is a water-soluble polymer made by polymeri zation of A/-vinylpyrrolidone. PVP exists in different grades, characterized by molecular weight, number and viscosity. Examples of suitable PVP grades are (see BASF, Tech nical Information 03-03-0730-e-09“Soluble Kollidon^® grades”, May 2013; Ashland, PVP Polyvinylpyrrolidone polymers, 2013): Kollidon^® 12 (M_w 2000-3000), Kollidon^® 12 PF (M_w 2000-3000), Kollidon^® 17 PF (M_w 7000-1 1000), Kollidon^® 25 (M_w 28000-34000), Kollidon^® 30 (Mw 44000-54000), PVP K-12 (M_w 4000-6000), PVP K-15 (M_w 6000-15000), PVP K-30 (M_w 40000-80000). Preferred are PVP grades having a M_w of between 2000 and 35000, more preferred between 2000 and 15000, particularly preferred between 5000 and 15000, in particular Kollidon^® 12 PF (Mw 2000-3000), Kollidon^® 17 PF (Mw 7000-1 1000), PVP K-15 (Mw 6000-15000), and very particularly Kollidon^® 17 PF (Mw 7000-1 1000). Mw means the average molecular weight in g/mol.

The amount of PVP employed in the composition according to the invention may vary in a narrow range. The composition according to the invention comprises preferably 0.1 to 1.0 % (w/v) of PVP, more preferably 0.1 to 0.75 % (w/v), even more preferably 0.25 to

0.5 % (w/v). Such mass concentrations relate to amounts of 1.0 to 10 mg/ml_, 1.0 to 7.5 mg/ml_ and 2.5 to 5 mg/ml_, respectively.

The pH of the inventive composition is adjusted with aqueous hydrochloric acid (HCI) to be between 3 and 4. Preferably the composition has a pH value of pH 4.

In one embodiment the composition according to the invention comprises

(a) 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6-(pyrro- lidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate (Neladenoson bialanate, I), preferably in the form of the hydrochloride (II) or mono-hydrochloride (lla),

(b) a chemically modified cyclodextrin selected from HP3CD and SBE3CD,

(c) a polyvinylpyrrolidone polymer having a Mw of between 5000 and 15000, and (d) water.

In another embodiment the composition according to the invention comprises

(a) 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6-(pyrro- lidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate (Neladenoson bialanate, I), preferably in the form of the hydrochloride (II) or mono-hydrochloride (lla) in an amount of 1 to 5 mg/ml_ [values calculated based on the compound of formula (I)],

(b) a chemically modified cyclodextrin selected from HP3CD and SBE3CD in an amount of 10 to 40 % (w/v),

(c) a polyvinylpyrrolidone polymer having a M_w of between 5000 and 15000 in an amount of 0.1 to 0.5 % (w/v), and

(d) water.

In another embodiment the composition according to the invention comprises

(a) 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6-(pyrro- lidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate (Neladenoson bialanate, I), preferably in the form of the hydrochloride (II) or mono-hydrochloride (lla) in an amount of 2.5 mg/ml_ [value calculated based on the compound of formula (I)],

(b) a chemically modified cyclodextrin, which is HP3CD, in an amount of 40 % (w/v), (c) a polyvinylpyrrolidone polymer having a Mw of between 7000 and 1 1000 (employed as Kollidon^® 17 PF) in an amount of 0.5 % (w/v), and

(d) water.

In the context of this invention“% (w/v)” defines the mass concentration of a component in percent within a composition, wherein w means the mass (measured in g, mg etc.) of the component employed, and v means the final volume (measured in L, ml_ etc.) of the composition.

Another subject of the invention is a method for preparing the compositions according to the invention. Thereby, the excipients are mixed until a clear solution is obtained. In a next step the pH is adjusted to the desired value. Then Neladenoson bialanate is added to the solution and is dissolved subsequently. Finally water is added up to the desired volume.

Typically the cyclodextrin is dissolved in water at the desired concentration and the pH- value is adjusted using HCI to the desired value (e.g. to pH 4 or pH 3). In the next step the polyvinylpyrrolidone polymer is added at the desired concentration. Subsequently the pH- value is adjusted again. Thereafter Neladenoson bialanate (mainly in form of the hydrochlo ride or mono-hydrochloride) is added at the desired concentration (e.g. 2.5 mg/ml_, calcula tion based on compound of formula (I)). Subsequently the pH value is adjusted again.

The compositions of the invention may be administered using an injector, a pump, a sy ringe, or any other devices/infusion devices known in the art as well as by gravity. A nee- die or a catheter may be used for introducing the formulations of the present invention in to the body of a patient via certain parenteral routes.

In another aspect the composition according to the invention, if prepared in large scale, can be used as aqueous bulk solution suitable for lyophilisation and a method of prepar ing the same.

In the light of the specific technical problems in formulating Neladenoson bialanate, it was surprisingly found that a manufacturing process was achieved which results in an aque ous bulk solution of Neladenoson bialanate which is suitable for direct lyophilisation.

It was also surprisingly found that after reconstitution of the lyophilisate, a solution was achieved which is suitable for direct application or dilution.

Further, the present invention is also based on the unexpected finding that the lyophilisate comprising Neladenoson bialanate prevents chemical and physical instability during long term storage. The term“lyophilized” as used herein means that the composition has been lyophilized. During lyophilisation, the liquid formulation is frozen and the solutes are separated from the solvent. The solvent is removed by sublimation (i.e., primary drying) and next by de sorption (i.e., secondary drying). Lyophilisation results in a cake or powder which can be stored over a long time period. Prior to administration, the lyophilized composition is re constituted in a suitable solvent, particularly water for injection.

The term“reconstituted solution/formulation” as used herein refers to such a lyophilized composition after adding a suitable diluent.

One aspect of the present invention relates to a method of preparing a stable, Neladenoson bialanate-containing aqueous bulk solution suitable for lyophilisation and for therapeutic applications, said bulk solution comprising:

• Neladenoson bialanate,

• a chemically modified cyclodextrin,

• a polyvinylpyrrolidone polymer, and

· water,

said bulk solution having a pH of between 3 and 4,

said method comprising:

• charging a suitable vessel with water;

• dissolving the chemically modified cyclodextrin, optionally by stirring, at the desired concentration (e.g. HP3CD at 400 mg/mL);

• adjusting the pH value to a pH of 3 to 4 with an appropriate amount of aqueous HCI;

• dissolving the polyvinylpyrrolidone polymer, optionally by stirring, at the desired concentration (e.g. PVP“17 PF” at 10 mg/mL);

· adjusting the pH value to a pH of 3 to 4 with an appropriate amount of aqueous

HCI;

• transferring Neladenoson bialanate, preferably in the form of a hydrochloride, in particular a monohydrochloride, at the desired concentration (e.g. 2.5 mg/mL) and allowing said Neladenoson bialanate to dissolve, optionally by stirring;

· adjusting again the pH value to a pH of 3 to 4 with an appropriate amount of aque ous HCI;

• adding water to reach the final weight; thus providing a stable, Neladenoson bialanate-containing aqueous bulk solution suitable for direct lyophilisation and therapeutic applications; and

• filling the thus-formed bulk solution in one or more vials.

Said method is hereinafter referred to as“manufacturing process” and said bulk solution is hereinafter referred to as“bulk solution”.

Said manufacturing process provides a stable, Neladenoson bialanate-containing aque ous bulk which contains Neladenoson bialanate at concentrations which are sufficiently high for their direct lyophilisation of the therapeutic dose in one or two vials, in particular one vial, which is of tolerated pH and which is suitable for therapeutic applications.

In another aspect the invention relates to a method of lyophilizing the Neladenoson bi alanate-containing bulk solution said lyophilisation method including freezing, annealing, primary and secondary drying steps.

In another aspect the invention relates to the a stable, lyophilized, Neladenoson bialanate containing solid, in particular powder or cake, in one or two containers, particularly one sealed container, (hereinafter referred to as“lyophilisate”).

Such lyophilized, Neladenoson bialanate-containing solid, in particular powder or cake, suitable for dilution and for therapeutic applications, comprises:

• Neladenoson bialanate,

• a chemically modified cyclodextrin, and

· a polyvinylpyrrolidone polymer.

said solid, in particular powder or cake, having a pH of between 3 and 4 when reconsti tuted with a suitable diluent, such as sterile water for injection or 5% glucose solution or 5% mannitol solution.

The lyophilized solid, in particular powder or cake, may further contain some remaining water. The remaining water is between 0 and 5 % (m/m), preferably between 0 and 2 % (m/m), further preferably between 0 and 0.5 % (m/m), further preferably at maximum 0.1 % (m/m).

Said stable, lyophilized, Neladenoson bialanate-containing solid, in particular powder or cake, once reconstituted into a reconstituted solution, is suitable for direct therapeutic appli- cations, or for further dilution with a suitable diluent, such as sterile water for injection or 5% glucose solution or 5% mannitol solution, and in turn for further therapeutic applications. In another aspect the invention relates to a method of reconstituting (“reconstituting method”) said lyophilisate and the reconstituted solution obtained therefrom (“reconstitut ed solution”).

The method of reconstituting a stable, lyophilized, Neladenoson bialanate-containing sol- id, in particular powder or cake, suitable for dilution and for therapeutic applications, com prises:

• adding to said lyophilisate in a suitable container, particularly a sealed container, particularly an injection vial, a diluent, such as sterile water for injection or 5% glu cose solution or 5% mannitol solution;

Said method is hereinafter referred to as“reconstitution method”.

The present invention further relates to a stable, reconstituted, Neladenoson bialanate- containing solution suitable for further dilution and for therapeutic applications, said re constituted solution comprising:

• Neladenoson bialanate, particularly in an amount suitable as a therapeutic dose, in one or two containers, particularly one sealed container,

• a chemically modified cyclodextrin,

• a polyvinylpyrrolidone polymer, and

• water,

said reconstituted solution having been prepared with a suitable diluent, such as sterile water for injection or 5% glucose solution or 5% mannitol solution,

said reconstituted solution having a pH of between 3 and 4.

The invention is explained further by means of the following examples.

Examples

A. Stability of Neladenoson bialanate in solution containing cyclodextrin

Solutions were manufactured by dissolving HP3CD at the respective concentration rang ing from 50 mg/ml_ to 400 mg/ml_ in water and adjusting pH using HCI. As a reference pH adjusted water was used. Neladenoson bialanate mono-hydrochloride (I la) was subse quently added at the respective concentration ranging from 0.625 mg/ml_ to 40 mg/ml_ [values calculated based on the compound of formula (I)]. The solution was then stirred for 5 days. Samples were taken in the beginning and after 24 h, 48 h and 120h. Hydroly sis of Neladenoson bialanate to Neladenoson was observed by determining the Neladenoson concentration at the respective time points. The experiments were carried out at room temperature. Analytical Method C was used to determine the Neladenoson (III) concentrations. The results are summarized in Table A.

Table A

^* Comparative examples

Without addition of cyclodextrin Neladenoson bialanate is rapidly hydrolyzed. With in creasing amount of cyclodextrin, Neladenoson bialanate hydrolysis is decreased.

B. Solubility of Neladenoson (III)

Solutions were manufactured by dissolving HRbOϋ or SBE3CD at the respective concen tration ranging from 50 mg/ml_ to 400 mg/ml_ in water and adjusting pH to either 3 or 4 using HCI. As a reference pH adjusted water was used. Neladenoson (III) was subse quently added to the solution in excess (~10 pg/mL).

The results are summarized in Table B (see Examples B-1 to B-20).

In addition solutions were manufactured by dissolving HRbOϋ at 400 mg/ml_ in water and PVP was added at concentrations between 0.001 and 1 mg/ml_. The pH was adjusted to 4 using HCI. The PVP used was either Kollidon^® 12 PF (M_w 2000-3000) or Kollidon^® 17 PF (Mw 7000-1 1000).

The results are summarized in Table B (see Examples B-21 to B-36).

The experiments were carried out at room temperature. To determine the Neladenoson (III) solubility Analytical Method B was used for Examples B-1 to B-20, and Analytical Method A was used for Examples B-21 to B-36.

Table B

^* Comparative examples

t“12 PF” = Kollidon^® 12 PF (M_w 2000-3000);“17 PF” = Kollidon^® 17 PF (M_w 7000-1 1000) not quant. = not quantifiable

In solutions containing up to 50 mg/ml_ cyclodextrin, Neladenoson (III) solubility was be- low the quantification limit of the respective analytical method. In solutions containing 100 - 400 mg/ml_ cyclodextrin, Neladenoson (III) solubility increased with increasing cyclo dextrin concentration.

By adding PVP“12 PF” to an aqueous solution containing 400 mg/ml_ HP3CD at pH 4, Neladenoson (III) solubility in solution further increased surprisingly reaching a maximum value when PVP“12 PF” is added at concentration between 1 and 5 mg/mL.

The same observation was made for solutions containing PVP“17 PF” instead. For PVP “17 PF” the overall solubility of Neladenoson (III) was even higher compared to solutions containing PVP“12 PF”. The maximum Neladenoson (III) solubility was also reached for PVP“17 PF” concentrations between 1 and 5 mg/mL.

C. Preparation of a Neladenoson bialanate bulk solution for lyophilisation

The solution was manufactured by dissolving HP3CD in water at 400 mg/mL. PVP“17 PF” (Kollidon^® 17 PF (Mw 7000-1 1000)) was dissolved at 0, 5 and 10 mg/mL, respectively.

The pH-value was adjusted to 4 using HCI. Neladenoson bialanate mono-hydrochloride (lla) was subsequently added at 2.5 mg/mL [value calculated based on the compound of formula (I)]. The pH-value was again adjusted to 4 using HCI. Water was added to reach the final weight. The solution was investigated with regard to Neladenoson (III) formation over a period of 48 hours. The experiments were carried out at room temperature. Analyt ical Method A was used to determine the Neladenoson (III) concentrations. The results are summarized in Table C.

Table C

^* Comparative examples All solutions showed a similar behavior with regard to Neladenoson bialanate stability after 48 hours.

D. Stability of Neladenoson bialanate after reconstitution

The solution was manufactured by dissolving HP3CD in water at 400 mg/mL PVP“17 PF” (Kollidon^® 17 PF (M_w 7000-1 1000)) was dissolved at 0, 5 and 10 mg/mL, respectively.

The phi-value was adjusted to 4 using HCI. Neladenoson bialanate mono-hydrochloride (lla) was subsequently added at 2.5 mg/mL [value calculated based on the compound of formula (I)]. The pH-value was again adjusted to 4 using HCI. Water was added to reach the final weight. The solution was filtered through a 0.2 pm filter membrane, filled into vi- als and lyophilized. The lyophilisation was carried out according to the following Lyophi- lisation Scheme :

Lyophilisation Scheme

The lyophilisate was reconstituted adding 7.2 mL sterile water for injection to the lyophi- lisate via needle and syringe. The stability of Neladenoson bialanate (I) was investigated over a period of 48 hours after reconstitution. The experiments were carried out at room temperature. Analytical Method A was used to determine the Neladenoson (III) concen trations. The results are summarized in Table D. Table D

^* Comparative examples

All solutions showed a similar behavior with regard to Neladenoson bialanate stability after 48 hours.

E. Stability of lyophilisate at pH 4.0

The lyophilisate was manufactured by dissolving HP3CD in water at 400 mg/mL PVP “17 PF” (Kollidon^® 17 PF (M_w 7000-1 1000)) was dissolved at 5 mg/mL. The phi-value was adjusted to 4 using HCI. Neladenoson bialanate mono-hydrochloride (I la) was subse quently added at 2.5 mg/mL [value calculated based on the compound of formula (I)]. The pFI-value was again adjusted to 4 using HCI. Water was added to reach the final weight. The solution was filtered twice through a 0.2 pm filter membrane, filled into vials and ly- ophilized. The lyophilisation was carried out according to the Lyophilisation Scheme shown in Example D above.

The resulting lyophilisate was stored at different conditions {6 , 25Ό/60% relative hu- midity (RH), 30Ό/75% RH and 40Ό/75% RH). Analytic al Method D was used to deter mine the Neladenoson (III) concentrations. The results after reconstitution are summa rized in Table E.

Table E

F. Stability of lyophilisate at pH 3.0

The lyophilisate was manufactured by dissolving HP3CD in water at 400 mg/mL PVP “17 PF” (Kollidon^® 17 PF (Mw 7000-1 1000)) was dissolved at 5 mg/mL. The phi-value was adjusted to 3 using HCI. Neladenoson bialanate mono-hydrochloride (I la) was subse quently added at 2.5 mg/mL [value calculated based on the compound of formula (I)]. The pH-value was again adjusted to 3 using HCI. Water was added to reach the final weight. The solution was filtered twice through a 0.2 pm filter membrane, filled into vials and ly- ophilized. The lyophilisation was carried out according to the Lyophilisation Scheme shown in Example D above.

The resulting lyophilisate was stored at different conditions (60, 250/60% relative hu midity (RH), 300/75% RH and 400/75% RH). Analytic al Method D was used to deter mine the Neladenoson (III) concentrations. The results after reconstitution are summa rized in Table F. Table F

Analytical Methods to determine Neladenoson (III)

Method A: Concentrations of Neladenoson were assessed by reversed-phase high perfor mance liquid chromatography (HPLC) on an Agilent system following appropriate dilution of the samples. 3.0 pi of sample solution was then injected on a Zorbax SB-CN (50 mm c 2.1 mm, 1 .8 pm) column (Agilent) kept at a temperature of 40Ό ( flow rate of 1.0 ml/min). Sam ples were analyzed using a 20-80 % (v/v) solvent B gradient over 10.0 min with the mobile phases composed of solvent A (H2O pH 2.4 (pH adjusted with phosphoric acid)) and sol vent B (acetonitrile). Neladenoson were assayed at a wavelength of 220 nm.

Method B: Concentrations of Neladenoson were assessed by reversed-phase high per formance liouid chromatography (HPLC) on an Agilent system following appropriate dilu tion of the samples. 1 .0 pi of sample solution was then injected on an Ascentis Express C18 (50 mm c 2.1 mm, 2.7 pm) column (Supelco) kept at a temperature of QOΌ ( flow rate of 1 .6 ml/min). Samples were analyzed using a 29-55 % (v/v) solvent B gradient over 4.5 min with the mobile phases composed of solvent A (10 mmol aqueous phosphoric ac id) and solvent B (acetonitrile). Neladenoson were assayed at a wavelength of 300 nm. Method C: Concentrations of Neladenoson were assessed by reversed phase HPLC with UV-detection at 300 nm on an Agilent system. The stationary phase is a YMC-Triart C18 HPLC column (100 mm x 3.0 mm, 1 .9 pm particle size). For peak separation, gradient elution with a flow rate of 0.6 mL/min is chosen using an ammonium acetate buffer (ad- justed to approximately pH 6 with acetic acid) as mobile phase A, and acetonitrile modi fied with 5 % methanol as mobile phase B. The applied gradient profile contains two steps running from 53 % to 70 % mobile phase B within 30 min and from 70 % to 75 % mobile phase B within another 10 min, followed by a 5 min isocratic phase at 75 % mobile phase B. The column temperature is kept at 20 an d the injection volume is 3.0 pL. Ap- propriate sample solutions are prepared by dissolving samples in a mixture of phosphate buffer (pH around 2.4, adjusted with phosphoric acid) / acetonitrile (1/1 ; V/V).

Method D: Concentrations of Neladenoson were assessed by reversed phase HPLC with UV-detection at 300 nm using a cell with 60 mm optical path length on an Agilent UHPLC system. The stationary phase is an YMC-Triart C18 HPLC column (100 mm x 3.0 mm, 1.9 pm particle size). For peak separation, gradient elution with a flow rate of 0.6 mL/min is chosen using an ammonium acetate buffer (adjusted to pH 6.1 with acetic acid) as mo bile phase A, and acetonitrile modified with 5 % methanol as mobile phase B. The applied gradient profile contains three steps running from 53 % to 61.5 % mobile phase B within 15 min and from 61.5 % to 85 % mobile phase B within another 15 min. The column tem- perature is kept at 20 Ό and the injection volume is 5.0 pL. Appropriate sample solutions are prepared by dissolving reconstituted samples in a mixture of 10 mmol/L aqueous phosphoric acid / acetonitrile (1/3; V/V).

Claims

1 . Composition comprising

(a) 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6- (pyrrolidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate (Neladenoson bialanate),

(b) a chemically modified cyclodextrin,

(c) a polyvinylpyrrolidone polymer, and

(d) water.

2. The composition as claimed in claim 1 , wherein Neladenoson bialanate is applied in as hydrochloride (II) or mono-hydrochloride (lla).

3. The composition as claimed in claim 1 or 2, wherein the chemically modified cy clodextrin is selected from hydroxypropyl derivatives of 3CD and yCD (i.e. HRbOϋ and HPyCD), the hydroxyethyl derivative of 3CD (HE3CD), the methylated b- cyclodextrin (Mbqϋ), the dimethyl^-cyclodextrin (DIV^CD), the randomly methylated bqϋ (RIV^CD), the permethylated b-cyclodextrin (RMbOϋ), the sulfobutylether of bqϋ and yCD (i.e. bBEbOϋ and SBEyCD), and the so-called branched CDs such as maltosyl^CD (Q2bOO) and glucosyl^-cyclodextrin (G^CD).

4. The composition as claimed in any of claims 1 to 3, wherein the polyvinylpyrrolidone polymer has a M_w of between 2000 and 35000.

5. The composition as claimed in any of claims 1 to 4 comprising

(a) 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6- (pyrrolidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate (Neladenoson bialanate, I) in the form of the hydrochloride (II) or mono-hydrochloride (lla),

(b) a chemically modified cyclodextrin selected from HRbOϋ and bBEbOϋ, (c) a polyvinylpyrrolidone polymer having a Mw of between 5000 and 15000, and

(d) water.

6. The composition as claimed in claim 5 comprising

(a) 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6- (pyrrolidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate (Neladenoson bialanate, I), preferably in the form of the hydrochloride (II) or mono- hydrochloride (lla) in an amount of 1 to 5 mg/ml_[values calculated based on the compound of formula (I)],

(c) a polyvinylpyrrolidone polymer having a Mw of between 5000 and 15000 in an amount of 0.1 to 0.5 % (w/v), and

(d) water.

7. The composition as claimed in claim 5 comprising

(a) 2-{4-[2-({[2-(4-chlorophenyl)-1 ,3-thiazol-4-yl]methyl}sulfanyl)-3,5-dicyano-6- (pyrrolidin-1 -yl)pyridin-4-yl]phenoxy}ethyl-L-alanyl-L-alaninate (Neladenoson bialanate, I), preferably in the form of the hydrochloride (II) or mono hydrochloride (lla) in an amount of 2.5 mg/ml_ [values calculated based on the compound of formula (I)],

(b) a chemically modified cyclodextrin, which is HRbOϋ, in an amount of 40 % (w/v),

(c) a polyvinylpyrrolidone polymer having a M_w of between 7000 and 1 1000 (em ployed as Kollidon^® 17 PF) in an amount of 0.5 % (w/v), and

(d) water.

8. A process for preparing the compositions according to any of claims 1 to 7, charac- terized in that the cyclodextrin is dissolved in water at the desired concentration, the pH-value is adjusted using HCI to the desired value, followed by addition of the pol yvinylpyrrolidone polymer with subsequent adjustment of the pH-value, and finally addition of Neladenoson bialanate (mainly in form of the hydrochloride or mono hydrochloride) at the desired concentration.

9. Use for of the composition according to claims 1 to 7 for treatment the prevention and/or treatment of cardiovascular disorders.

10. Use according to claim 9, where the composition is applied parenteral.

1 1 . Neladenoson bialanate containing aqueous bulk solution suitable for lyophilisation comprising a composition according to any of claims 1 to 7.

12. A method of preparing a stable, Neladenoson bialanate-containing aqueous bulk so lution suitable for lyophilisation and for therapeutic applications, said bulk solution comprising a composition according to any of claims 1 to 7, said bulk solution having a pH of between 3 and 4,

said method comprising:

• charging a suitable vessel with water;

• dissolving the chemically modified cyclodextrin, optionally by stirring, at the desired concentration (e.g. HP3CD at 400 mg/ml_);

• adjusting the pH value to a pH of 3 to 4 with an appropriate amount of aque ous HCI;

• dissolving the polyvinylpyrrolidone polymer, optionally by stirring, at the de sired concentration (e.g. PVP“17 PF” at 10 mg/ml_);

• transferring Neladenoson bialanate, preferably in the form of a hydrochloride, in particular a monohydrochloride, at the desired concentration (e.g. 2.5 mg/ml_) and allowing said Neladenoson bialanate to dissolve, optionally by stir ring;

• adjusting again the pH value to a pH of 3 to 4 with an appropriate amount of aqueous HCI;

• adding water to reach the final weight;

thus providing a stable, Neladenoson bialanate-containing aqueous bulk solution suitable for direct lyophilisation and therapeutic applications; and

• filling the thus-formed bulk solution in one or more vials.

13. A stable, lyophilized, Neladenoson bialanate containing solid, in particular powder or cake, in one or two containers, particularly one sealed container,

wherein such lyophilized solid comprises:

• Neladenoson bialanate,

• a chemically modified cyclodextrin, and

• a polyvinylpyrrolidone polymer.

14. A method of reconstituting a stable, lyophilized, Neladenoson bialanate-containing solid, in particular powder or cake, suitable for dilution and for therapeutic applica tions, according to Claim 13 comprising adding to said lyophilisate in a suitable container, particularly a sealed container, particularly an injection vial, a diluent, such as sterile water for injection.

15. A stable, reconstituted, Neladenoson bialanate-containing solution suitable for fur ther dilution and for therapeutic applications, said reconstituted solution comprising a composition according to any of claims 1 to 7.