EA044509B1 - THERAPEUTIC, LYOPHILIZED, COPANLISIB-CONTAINING SOLID (VERSIONS) AND DEVICE CONTAINING THEREFROM - Google Patents

Description

The present invention relates to a therapeutic, lyophilized, copanlisib-containing solid containing: copanlisib, in a therapeutically effective amount, one or more pH adjusters, a buffering agent containing a weak acid such as citric acid, and a strong base such as sodium hydroxide, and a bulking agent which is mannitol, wherein said therapeutically effective amount is 30, 45, 60 or 80 mg of copanlisib or 68.4 mg;

to a device that is a container containing said lyophilized copanlisib-containing solid.

Copanlisib is a novel intravenous class I phosphatidylinositol 3-kinase (PI3K) inhibitor with kinase inhibitory activity primarily against the PI3K-a and PI3K-8 isoforms that are expressed in malignant B cells. The PI3K pathway is involved in cell growth, survival and metabolism, and its dysregulation plays an important role in non-Hodgkin's lymphoma (NHL).

Copanlisib exhibits broad spectrum activity against tumors of multiple histological types, both in vitro and in vivo.

Copanlisib can be synthesized in accordance with the methods set forth in international patent application PCT/EP2003/010377, published as WO 04/029055 A1 dated April 08, 2004, (which is incorporated herein by reference in its entirety), at ss. 26 ff.

Copanlisib is disclosed in international patent application PCT/US 2007/024985, published as WO 2008/070150 A1 dated June 12, 2008, (which is incorporated herein by reference in its entirety) as the compound of Example 13: 2-amino-N- [7-methoxy-8-(3-morpholin-4-ylpropoxy)-2,3dihydroimidazo[1,2-c]quinazolin-5-yl]pyrimidin-5-carboxamide, which is a free base. Copanlisib free base can be synthesized in accordance with the methods given in WO 2008/070150, ss. 9 ff., and on ss. 42 ff. Biological test data for compounds of formula (I) are given in WO 2008/070150 at cc. 101-107.

Copanlisib dihydrochloride, which is 2-amino-N-[7-methoxy-8-(3-morpholin-4ylpropoxy)-2,3-dihydroimid-azo[1,2-c]quinazolin-5-yl]pyrimidin-5- carboxamide dihydrochloride, (hereinafter referred to as copanlisib dihydrochloride), is disclosed in international patent application PCT/EP2012/055600, published as WO 2012/136553 dated October 11, 2012, (which is incorporated herein by reference in its entirety) as the compound from examples 1 and 2: 2-aminoN-[7-methoxy-8-(3-morpholin-4-ylpropoxy)-2,3-dihydroimidazo[1,2-c]quinazolin-5-yl]pyrimidin-5carboxamide dihydrochloride: it can be synthesized in accordance with the methods given in the above examples 1 and 2.

Copanlisib free base and copanlisib dihydrochloride can also be synthesized in accordance with methods published in international patent application PCT/EP2015/075789, published as WO 2016/071435 dated May 12, 2016, (which is incorporated herein by reference in its entirety).

Copanlisib dihydrochloride exists in an amorphous form or in one of three crystalline forms, which are hydrate I, II and III

. 2 HCI. x H ₂ O

In the context of the present invention, unless otherwise specified, the term copanlisib or drug substance refers to copanlisib in the form of a free base, or salt, such as a hydrochloride, in particular a monohydrochloride or dihydrochloride, or a tosylate, for example, this form may exist as a hydrate, anhydrate or in amorphous form.

The compound used as a drug substance in the present invention is copanlisib dihydrochloride.

Specific technical issues associated with the preparation of copanlisib

Copanlisib exhibits increased solubility in aqueous solutions with decreasing pH, which, however, is associated with a decrease in stability and, in addition, low local tolerance of this potential composition.

Copanlisib dihydrochloride exhibits pH-dependent solubility: it is readily soluble in 0.1 M hydrochloric acid, phosphate buffer pH 1, phosphate buffer pH 2, citrate buffer pH 3. It is soluble in citrate buffer pH 4. It is very slightly soluble in citrate buffer pH 5. It is practically insoluble in phosphate buffer pH 6 and phosphate buffer pH 7.

According to the IUPAC definition, solubility is an analytical composition

- 1 044509 of a saturated solution, expressed as the proportion of the specified solute in the specified solvent.

Solubility is the property of a solid, liquid or gaseous chemical substance, called a solute, to dissolve in a solid, liquid or gaseous solvent. The solubility of a substance mainly depends on the physical and chemical properties of the solute and solvent, as well as the temperature, pressure and pH of the solution. The degree of solubility of a substance in a particular solvent is measured as the saturation concentration, where adding more solute does not increase the concentration of the solution and begins to precipitate the excess amount of solute. The solubility of a substance is a completely different property from the rate of dissolution, which depends on how quickly it dissolves.

Under certain conditions, equilibrium solubility can be exceeded to produce a so-called supersaturated solution, which is metastable.

Supersaturated solutions will crystallize under certain conditions (see Crystallization of Molecular Systems from Solution: Phase Diagrams, Supersaturation, and Other Basic Concepts, Chemical Society Reviews, RSC Publishing. 43: 2286). In a normal solution, once the maximum amount of solute has been dissolved, adding more solute will either cause the solute to precipitate and/or the solute not to dissolve at all (see Mechanism of Precipitation Formation During Spontaneous Crystallization from Supersaturated Aqueous Waters solutions; Chem. Review. 83: 343-364. 2014). In some cases, the solubility of a saturated solution is reduced by a change in temperature, pressure or volume, but a supersaturated state does not occur. In these cases, the solute simply precipitates. This is because a supersaturated solution is in a state of higher energy than a saturated solution.

It is important to note that solubility applies to all areas of chemistry, geochemistry, inorganic, physical, organic and biochemistry. In all cases, this will depend on the physical conditions (temperature, pressure and concentration) as well as the enthalpy and entropy directly associated with the solvents and solutes involved.

Traditional lyophilization is a complex process requiring careful balancing of product, equipment and processing technologies (see International Journal of Novel Trends in Pharmaceutical Sciences. 3(4). 2013). Apart from various advantages, lyophilization also has many disadvantages such as long processing time, aseptic processing, limitations in size and filling volume of suitable containers and associated costs. Since lyophilization is a complex process, certain experience and knowledge about the critical temperature of the composition/temperature of destruction of the composition and freeze-drying parameters are required. The structure of the lyophilisate briquette and the solid state of the active and inactive ingredients are influenced by both composition and processing parameters.

In the specific case of copanlisib, the required therapeutic dose of copanlisib is soluble only in large volumes of diluent or at a pH that is poorly tolerated during and after parenteral administration, i.e. during and after administration. Parenteral formulations should be designed to be isotonic and euhydric. Parenteral formulations should have a pH value that corresponds to the physiological pH at the site of application or, in the case of infusions, the physiological pH of the blood. Taking into account the buffering capacity of human tissues, organs and blood, small volume parenteral formulations with pH values of 4-8 are considered very acceptable.

As mentioned above, copanlisib exhibits pH-dependent solubility and pH-dependent instability: Copanlisib exhibits hydrolytic instability in various media such as water, pH 7 buffer and 0.1 M hydrochloric acid and 0.1 M sodium hydroxide solution.

In general, degradation of a given active pharmaceutical ingredient may occur due to chemical instability, resulting in the emergence of a new chemical entity. For example, chemical instability may result from hydrolysis and/or oxidation. Degradation of active pharmaceutical ingredients is an undesirable effect for pharmaceutical applications. The effectiveness or availability of the drug may change dramatically.

The active pharmaceutical ingredient may also be physically unstable. Physical instability of a solution may result from crystallization and/or precipitation when the solubility limit/solubility equilibrium of the active pharmaceutical ingredient is exceeded in the solution, which can lead to the formation of particles. The formation of particles in pharmaceutical preparations for parenteral use, in particular, makes the composition harmful for clinical use.

Therefore, copanlisib-containing bulk solutions used for further processing must demonstrate sufficient physical and chemical stability (retention time) to ensure formulation integrity. Ready-to-use solutions must also demonstrate sufficient physical and chemical stability (stability in use) to avoid harm upon final administration to the patient.

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As is known in the pharmaceutical formulation art, lyophilization, also known as freeze-drying, is a method of converting a liquid product into a dry solid product.

In general, lyophilization is defined as a stabilization process in which a product is frozen and then the water content is removed by sublimation.

The resulting lyophilized product must have an acceptable cake structure and sufficient stability (shelf life), short rehydration/reconstitution time and sufficient stability when used at the required temperature. One of the main disadvantages of lyophilized products is the fact that the stability of the drug depends both on its physical state and on the physical state of all components.

Before lyophilization, the active pharmaceutical ingredient, together with any excipients such as bulking agents, pH adjusters, cryoprotectants that may be present, for example, must be dissolved in a suitable solvent to form a solution (bulk solution). After freeze-drying, the resulting dry solid and the active ingredient therewith must be redissolved in a suitable diluent or solvent to form a solution (reconstituted solution). This reconstituted solution can then be administered to the patient directly or after further dilution.

As a prerequisite for lyophilization, a small filling volume is required to facilitate an efficient lyophilization process. Therefore, lyophilization is only suitable for active ingredients that provide high water solubility or a low therapeutic dose to reduce the number of vials per administration. Thus, in the case of high therapeutic doses, such as 60 mg for copanlisib, the required number of vials to administer depends on the solubility of the drug substance.

Mannitol is widely used as a bulking agent for lyophilized products. During lyophilization, mannitol has been demonstrated to crystallize as three common stable anhydrous polymorphs (i.e., α, β, and δ) or as mannitol hemihydrate (see Mechanism of precipitate formation during spontaneous crystallization from supersaturated aqueous solutions; Chem. Review 83: 343-364. 2014). Common stable anhydrous polymorphs of mannitol may also be called modification I (hereinafter referred to as mod. I or β), II (hereinafter referred to as mod. II or α) and III (hereinafter referred to as mod. III or δ) (see Journal of Pharmaceutical Sciences, Volume 89, Issue 4, April 2000, pp. 457-468 (A. Burger, J.-O. Henck, S. Hetz, J. M. Rollinger, A. A. Weissnicht, H. Stottner).

One of the reasons for the widespread use of mannitol is its tendency to crystallize from frozen aqueous solutions. This property should facilitate efficient freeze-drying and a physically stable, pharmaceutically attractive lyophilized solid.

Unlike many excipients (eg, sorbitol and disaccharides), mannitol has a strong tendency to crystallize from frozen aqueous solution, both when cooled and when reheated. The phenomenon of bottle destruction is a clear illustration of this trend. Mannitol has been observed to continue to crystallize after freeze-drying, especially when exposed to heat and moisture, indicating that the freeze-drying process may result in a partially amorphous and partially crystalline material (see Yu et al., Journal of Pharmaceutical Sciences v. 88, No. 2, February 1999).

Mannitol has been reported to have an adverse effect on the stability of lyophilized solid drugs. In Herman et al. reported that the rate of hydrolysis of methylprednisolone sodium succinate in the lyophilized solid state is significantly higher when mannitol is used as a bulking agent compared to an amorphous excipient such as lactose (see Journal of Pharmaceutical Sciences Vol. 87, No. 8, August 1998). This instability of the drug in the presence of mannitol was attributed, at least in part, to the ongoing crystallization of mannitol from a system that was initially only partially crystalline. Although amorphous mannitol can serve as a stabilizer for an active pharmaceutical ingredient (API) (see Izutsu et al., Chemical and Pharmaceutical Bulletin Vol. 42, No. 1, January 1994), the difficulty of maintaining mannitol in an amorphous state during lyophilization makes mannitol a poor choice as a stabilizer (See Pikal, 2002. Freeze drying. In: Swarbrick, J., Boylan, J. (Eds.), Encyclopedia of Pharmaceutical Technology. Marcel Dekker, New York, cc. 1807-1833).

There is still a need to better understand the physical chemistry of freeze-drying mannitol-containing formulations to anticipate and avoid the adverse effects of mannitol on the physical and chemical stability of the lyophilized solid. One skilled in the art will appreciate that the physical state of mannitol in a lyophilized solid is influenced by both composition and processing parameters.

Taking into account the above, due to the specific technical problems associated with compiling

- 3 044509 copanlisib, the object of the present invention is to provide a pharmaceutically acceptable formulation of copanlisib that prevents chemical and physical instability;

a stable aqueous solution containing copanlisib, which is suitable for direct parenteral administration or further dilution;

a stable formulation containing the total therapeutic dose of copanlisib in a maximum of 1 or 2 suitable containers;

a manufacturing method that produces a pharmaceutically acceptable formulation of copanlisib for parenteral administration.

Applicant has knowledge that such compositions containing copanlisib and such production methods for the manufacture of compositions containing copanlisib are unknown.

A pharmaceutically acceptable formulation of copanlisib that prevents chemical and physical instability can be prepared by lyophilization. An aqueous solution containing copanlisib, which is suitable for direct parenteral administration or further dilution, requires a pH that is tolerated at the site of administration and does not disturb the physiological pH of the blood. Regarding the low solubility of copanlisib at tolerable/physiological pH, large volume lyophilization would be necessary to cover the total therapeutic dose. Since lyophilization of large volume containers is not feasible, this approach is expected in the art to result in lyophilization of multiple containers to cover a therapeutic dose of copanlisib.

Therefore, increased solubility of copanlisib in aqueous solution is required to enable lyophilization of a formulation containing a total therapeutic dose in 1 or 2 vials.

In light of the particular technical problems associated with the formulation of copanlisib, it was unexpectedly and unexpectedly discovered, and this is the basis of the present invention, that a manufacturing method was discovered that results in an aqueous bulk solution of copanlisib that provides increased solubility of the drug and is suitable for direct lyophilization .

It was also unexpectedly found that after reconstitution of the lyophilisate, a solution was obtained which showed increased solubility of copanlisib and which was suitable for direct use or dilution.

In addition, the present invention is also based on the unexpected discovery that a lyophilisate containing copanlisib, a pH regulator and a bulking agent prevents chemical and physical instability during long-term storage.

In particular, it has been unexpectedly discovered that the method of manufacture in accordance with the present invention provides an aqueous copanlisib-containing bulk solution and that the reconstitution process even results in a copanlisib-containing reconstituted solution that exhibits increased solubility in the corresponding solvent, which is substantially higher than the expected solubility in this solvent according to the solubility data of copanlisib (see Examples section).

In addition, it was unexpectedly found that the bulk solution of copanlisib and the reconstituted solution do not exhibit physical instability (e.g. sedimentation) and provide sufficient residence time and stability in use despite the metastable state of the solution: this is very surprising in light of the prior art/common knowledge in this area.

In addition, it was unexpectedly found that the bulk solution of copanlisib does not exhibit crystallization/precipitation upon freezing and annealing during the lyophilization process, despite the metastable state of the solution: this is also very surprising in light of the prior art/general knowledge in the field.

In addition, it has surprisingly been found that a lyophilisate formulation containing copanlisib, a pH adjuster and a crystalline bulking agent prevents crystallization of the amorphous drug phase on the crystalline bulking agent during long-term storage: this is very surprising in light of the prior art/general knowledge in the art.

In addition, it was unexpectedly discovered that a lyophilisate formulation containing copanlisib, a pH adjuster and a crystalline bulking agent prevented crystallization of the amorphous drug phase on the crystalline bulking agent at elevated levels of residual moisture of the lyophilisate formulation: this is very unexpected in light of the prior art/general knowledge in the art. areas due to the metastable state of the drug substance in the composition.

As described above, one skilled in the art would not conclude that the solubility of copanlisib can be increased and maintained over several weeks in bulk and reconstituted solution forms and during lyophilization, including freezing and annealing, without solubilizing or stabilizing agents. (e.g. emulsifiers, polymers).

As described above, one skilled in the art would not conclude that the amorphous state of copanlisib achieved during lyophilization can be physically maintained during long-term storage as long as mannitol is present in crystalline form.

Accordingly, the present invention relates to a therapeutic, lyophilized, copanlisib-containing solid containing copanlisib, in a therapeutically effective amount, one or more pH adjusters, a buffering agent containing a weak acid such as citric acid, and a strong base such as hydroxide sodium, and a bulking agent which is mannitol, wherein said therapeutically effective amount is 30, 45, 60 or 80 mg of copanlisib or 68.4 mg;

to a device that is a container containing said lyophilized copanlisib-containing solid. Said therapeutic, stable, lyophilized, copanlisib-containing solid can be obtained by a process for preparing a stable, copanlisib-containing, aqueous bulk solution suitable for lyophilization and for therapeutic use (hereinafter referred to as a manufacturing method). Stable copanlisib-containing bulk solution (hereinafter referred to as bulk solution) of increased solubility, directly suitable for lyophilization;

a method for lyophilizing said bulk solution (hereinafter designated as a lyophilization method).

Said stable, lyophilized, copanlisib-containing solid is, in particular, a powder or briquette, in particular contains a therapeutic dose of copanlisib in one or two containers, in particular, in one sealed container (hereinafter referred to as lyophilisate) and can be prepared . The invention describes a method for the recovery of said lyophilisate (hereinafter referred to as a recovery method). A stable, reconstituted solution containing copanlisib with increased solubility is suitable for further dilution and therapeutic use (hereinafter referred to as reconstituted solution).

Definitions used in this text

As used herein, the term pharmaceutical composition or composition refers to a preparation that is in a form that allows the biological activity of the active ingredient contained therein to be effective and that does not contain additional components that are unacceptably toxic to the subject to whom the composition is administered .

As used herein, the term pH adjuster refers particularly, but not exclusively, to a buffered solution whose pH changes only slightly after the addition of acidic or basic substances. Buffered solutions contain a mixture of a weak acid and its corresponding base, or a weak base and its corresponding acid, respectively. The term buffering agent as used herein refers to a mixture of one or more of the above acids and bases. pH adjusters that can be used in the present invention are, for example, buffers such as citric acid or a salt thereof, acetic acid or a salt thereof, or phosphoric acid or a salt thereof, or an inorganic acid such as, for example, hydrochloric acid , boric acid, carbonic acid, bicarboxylic acid or amino acid or organic acid such as monocarboxylic, oxocarboxylic or polycarboxylic acid, or a base such as, for example, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate. Specifically, the buffering agent is citric acid and NaOH. Most preferably, the buffering agent contains a mixture of citric acid and NaOH. In the context of the present invention, the term citric acid refers to the anhydrous form or its hydrate or salt thereof, such as sodium citrate, for example, unless specifically stated otherwise.

As used herein, the term suitable diluent/solvent refers in particular, but not exclusively, to aqueous sodium chloride 0.9%, glucose 5%, water for injection, Ringer's solution, mannitol 5%, etc.

As used herein, the term lyophilized means that the composition has been lyophilized. During lyophilization, the liquid formulation is frozen and the solutes are separated from the solvent. The solvent is removed by sublimation (ie, primary drying) and then desorption (ie, secondary drying). Lyophilization results in a cake or powder that can be stored for a long period of time. Before administration, the lyophilized composition is reconstituted in a suitable solvent, in particular an aqueous solution of sodium chloride.

As used herein, the term reconstituted solution/formulation refers to such a lyophilized composition after addition of a suitable diluent.

Lyophilization methods are well known in the art (for example, see Wang, International Journal of Pharmaceutics Vol. 203, 2000). Lyophilized compositions in accordance with the present invention were prepared by various lyophilization methods, as described in the Examples section below.

As used herein, the term fixed dose refers to the administration of 60 mg of copanlisib.

A therapeutic, lyophilized, copanlisib-containing solid can be prepared by preparing a bulk solution.

- 5 044509

A method for preparing a stable copanlisib-containing aqueous bulk solution suitable for lyophilization and for therapeutic use, wherein said bulk solution contains copanlisib, in particular in an amount suitable as a therapeutic dose, one or more pH regulators, in particular a buffering agent, which, in particular, contains a weak acid, such as, for example, citric acid, and a strong base, such as sodium hydroxide, for example, and a bulking agent, said bulk solution having a pH between 4 and 5 (inclusive), said method involves charging a suitable a vessel with water for injection;

transferring copanlisib in the form of a free base or salt, such as, for example, a hydrochloride, in particular a monohydrochloride or dihydrochloride, or tosylate, into said vessel;

causing said copanlisib to dissolve, optionally by stirring, for example at a pH between 2 and 3 (inclusive), thereby forming a solution containing copanlisib;

adding said buffering agent and said bulking agent to said copanlisib-containing solution;

causing said buffering agent and said bulking agent to dissolve, optionally, for example, by stirring;

adjusting the pH of the solution thus formed to a pH of 4 to 5 with an appropriate amount of sodium hydroxide solution, thereby forming a bulk solution;

filtering said bulk solution in a sterile manner, thereby providing a stable copanlisib-containing, aqueous bulk solution suitable for direct lyophilization and therapeutic applications; and filling the filtered bulk solution thus formed into one or more bottles.

Said copanlisib transferred into said vessel is copanlisib dihydrochloride.

Said method is hereinafter designated as a manufacturing method, and said bulk solution is hereinafter referred to as bulk solution.

This manufacturing method provides a stable aqueous bulk solution containing copanlisib, which gives increased solubility of copanlisib, i.e. which contains copanlisib in concentrations that are high enough to directly lyophilize a therapeutic dose in one or two vials, particularly one vial that has an acceptable pH level and is suitable for therapeutic use.

C stable, copanlisib-containing aqueous bulk solution of increased solubility, which is directly suitable for lyophilization, contains copanlisib, in particular in an amount suitable as a therapeutic dose, one or more pH regulators, in particular a buffering agent, which in particular contains a weak acid such as, for example, citric acid, and a strong base, such as, for example, sodium hydroxide and a bulking agent, said bulk solution has a pH between 4 and 5 (inclusive).

wherein said therapeutic dose is 30, 45, 60 or 80 mg of copanlisib.

In an embodiment, the therapeutic dose is 60 mg of copanlisib.

According to an embodiment, the pH regulator is a buffering agent.

According to an embodiment, the buffering agent is citric acid and sodium hydroxide.

According to an embodiment, said bulk solution containing copanlisib contains citric acid in an amount of 0.18-18.5% w/w, in particular 0.18-3.7% w/w, in particular 0. 37 w/w% of said bulk solution.

According to an embodiment, said bulk solution containing copanlisib contains sodium hydroxide in an amount of 0-0.8% w/w, in particular 0.3-0.5% w/w, in particular 0.4% w/w. /wt.% of the specified bulk solution.

According to an embodiment, said bulk solution containing copanlisib contains sodium hydroxide in an amount of 0.01-0.8% w/v, in particular 0.3-0.5% w/v, in particular 0.4 w/v% of said bulk solution.

According to an embodiment, said bulking agent is mannitol.

According to an embodiment, said bulk solution containing copanlisib contains mannitol in an amount of 3.0-23.1% w/w, in particular 3.0-15.4% w/w, in particular 7.7 w/w% of said bulk solution.

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According to an embodiment, said bulk solution containing copanlisib contains said copanlisib free base in an amount of 0.5-15.2% w/w, in particular 1.0-7.6% w/w, in particular 3 .8 wt./wt.% of the specified bulk solution.

In accordance with an embodiment, in said bulk solution containing copanlisib, said therapeutic dose of copanlisib is present in an amount of 3.8% w/w of said bulk solution, said citric acid is present in an amount of 0.37% w/w of said bulk solution. solution, said sodium hydroxide is present in an amount of 0.4% w/w of said bulk solution and said mannitol is present in an amount of 7.7% w/w of said bulk solution, said copanlisib containing solution has a pH between 4 and 5 (inclusive).

According to an embodiment, in said bulk solution containing copanlisib:

said therapeutic dose of copanlisib is present in an amount of 68.4 mg, said citric acid is present in an amount of 6.6 mg, said sodium hydroxide is present in an amount of 7.2 mg and said mannitol is present in an amount of 136.8 mg, said bulk solution containing copanlisib has pH between 4 and 5 (inclusive), said bulk solution containing copanlisib has a volume of 1.71 ml.

In accordance with an embodiment, said bulk solution containing copanlisib contains copanlisib in an amount of 40 mg/ml.

In an embodiment, the bulk solution containing copanlisib contains 68.4 mg of copanlisib in a volume of 1.71 ml.

In an embodiment, said bulk solution containing copanlisib contains 60 mg of copanlisib in a volume of 1.5 ml.

In an embodiment, said bulk solution containing copanlisib is further diluted with a suitable diluent, such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution.

According to an embodiment, said bulk solution containing copanlisib, wherein:

said therapeutic dose of copanlisib is present in an amount of 68.4 mg, said citric acid is present in an amount of 6.6 mg, said sodium hydroxide is present in an amount of 7.2 mg and said mannitol is present in an amount of 136.8 mg, further diluted with a diluent that is a 0.9% aqueous solution of sodium chloride, in an amount of 4.4 ml, said bulk solution containing copanlisib has a pH between 4 and 5 (inclusive), said bulk solution containing copanlisib has a volume of 1.71 ml.

In an embodiment, said bulk solution containing copanlisib is further diluted with a suitable diluent, said suitable diluent being a 0.9% aqueous solution of sodium chloride, especially in a 100 ml volume.

In an embodiment, said bulk solution containing copanlisib is further diluted with a suitable diluent, said suitable diluent being a 0.9% aqueous solution of sodium chloride, especially in a 100 ml volume, which is contained in an infusion bag, especially made of polyethylene (PE). ), polypropylene (PP), polyvinyl chloride (PVC) or ethylene vinyl acetate (EVA).

Said bulk solution is hereinafter referred to as such.

A method for lyophilizing a stable, copanlisib-containing, aqueous bulk solution suitable for lyophilization and for therapeutic use, wherein said composition contains copanlisib, in particular in an amount suitable as a therapeutic dose, one or more pH regulators, in particular a buffering agent, which in particular , contains a weak acid, such as citric acid, and a strong base, such as sodium hydroxide, and a bulking agent,

- 7 044509 said bulk solution has a pH between 4 and 5 (inclusive), said lyophilization method includes the stages of freezing, annealing, primary and secondary drying;

Said method thus provides a stable, lyophilized, copanlisib-containing solid, in particular a powder or cake, suitable for reconstitution and for therapeutic use, said solid, in particular a powder or cake, comprising:

copanlisib, in particular in an amount suitable as a therapeutic dose, one or more pH regulators, in particular a buffering agent, which in particular contains a weak acid, such as, for example, citric acid, and a strong base, such as, for example, hydroxide sodium, and the bulking agent, said solid, in particular a powder or cake, has a pH between 4 and 5 (inclusive), when reconstituted with a suitable diluent such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution.

Said reconstitution method involves adding to said stable lyophilized solid, in particular a powder, in a suitable container, in particular in a sealed container, in particular in an injection vial, in particular in a 6 ml injection vial, a diluent such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, in particular 0.9% aqueous sodium chloride solution;

The stable lyophilized copanlisib-containing solid, in particular a powder or cake, after reconstitution into a reconstituted solution is suitable for further dilution and therapeutic use.

This method is hereinafter referred to as a lyophilization method. This lyophilization process provides a stable, lyophilized, copanlisib-containing solid, in particular a powder or cake, which has an acceptable cake structure and a short reconstitution time.

In accordance with a first aspect, the present invention relates to a stable, lyophilized, copanlisib-containing solid, in particular a powder or cake, suitable for reconstitution and for therapeutic use, wherein said solid, in particular the powder or cake contains copanlisib, in a therapeutically effective amount, one or more pH adjusters, which is a buffering agent containing a weak acid, such as citric acid, and a strong base, such as sodium hydroxide, and a bulking agent, which is mannitol, said solid, in particular a powder or briquette, has in particular, a pH between 4 and 5 (inclusive) when reconstituted with a suitable diluent such as sterile water for injection or sterile aqueous sodium chloride solution, in particular 0.9% aqueous sodium chloride solution, for example and said therapeutically effective amount is 30, 45, 60 or 80 mg copanlisib.

In accordance with an embodiment of the fourth aspect of the present invention, said therapeutically effective amount is 60 mg of copanlisib.

According to an embodiment of the first aspect of the present invention, said buffering agent is a citric buffer which gives a pH value of between 4 and 5 (inclusive), in particular 5, when reconstituted with a suitable diluent such as sterile water for injection or sterile aqueous sodium chloride solution , especially 0.9% aqueous sodium chloride solution, for example.

According to an embodiment of the first aspect of the present invention, wherein said copanlisib-containing solid, in particular a powder or briquette, contains citric acid in an amount of 0.7-75 wt./wt.%, in particular 1.5-30 wt. /wt.%, in particular 2.98 wt./wt.%, of the specified lyophilized, containing copanlisib solid.

According to an embodiment of the first aspect of the present invention, wherein said copanlisib-containing solid, in particular a powder or briquette, contains sodium hydroxide in an amount of 0-6 wt./wt.%, in particular 3.3-4.0 wt. /wt.%, in particular 3.7 wt./wt.%, from the specified lyophilized, containing copanlisib solid.

According to an embodiment of the first aspect of the present invention, wherein said copanlisib-containing solid, in particular a powder or briquette, contains sodium hydroxide in an amount of 0.01-6% w/w, in particular 3.3-4.0 w/w%, in particular 3.7 w/w%, of said lyophilized copanlisib-containing solid.

According to an embodiment of the first aspect of the present invention, wherein said copanlisib-containing solid, in particular a powder or briquette, contains mannitol in an amount of 15.4-82.1% w/w, in particular 30.8-82, 1% w/w, in particular 61.6% w/w, of said lyophilized copanlisib-containing solid.

According to an embodiment of the first aspect of the present invention, in which

- 8 044509 said copanlisib-containing solid, in particular a powder or briquette, contains said copanlisib free base in an amount of 7.7-92.4 wt./wt.%, in particular 7.7-61.6 wt./wt. %, in particular 30.8 wt./wt.% of the lyophilized solid containing copanlisib.

According to an embodiment of the first aspect of the present invention, wherein in said lyophilized copanlisib-containing solid, particularly powder or cake, copanlisib is present in an amount of 30.8 w/w% of said lyophilized copanlisib-containing solid, said citric acid the acid is present in an amount of 2.98 w/w% of said lyophilized copanlisib-containing solid, said sodium hydroxide is present in an amount of 3.7 w/w% of said lyophilized copanlisib-containing solid, and said mannitol is present in an amount of 61.6% w/w of said lyophilized copanlisib-containing solid, said lyophilized copanlisib-containing solid, particularly powder or cake, when reconstituted with a suitable diluent such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution, has a pH between 4 and 5 (inclusive).

In accordance with an embodiment of the first aspect of the present invention, in said lyophilized copanlisib-containing solid, in particular powder or briquette, copanlisib is present in an amount of 30, 45 or 60 mg, said citric acid is present in an amount of 5.8 mg, said sodium hydroxide is present in an amount of 6.3 mg and said mannitol is present in an amount of 120 mg, said lyophilized copanlisib-containing solid, in particular a powder or cake, when reconstituted with a suitable diluent such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, in Features 0.9% aqueous sodium chloride solution has a pH between 4 and 5 (inclusive).

According to an embodiment of the first aspect of the present invention, in said lyophilized copanlisib-containing solid, in particular powder or briquette, copanlisib is present in an amount of 80 mg, said citric acid is present in an amount of 7.7 mg, said sodium hydroxide is present in an amount of 8.4 mg and said mannitol is present in an amount of 160 mg, said lyophilized copanlisib-containing solid, in particular a powder or cake, when reconstituted with a suitable diluent such as, for example, sterile water for injection or a sterile aqueous solution of sodium chloride, especially 0.9 % aqueous sodium chloride solution has a pH between 4 and 5 (inclusive).

According to an embodiment of the first aspect of the present invention, said lyophilized copanlisib-containing solid, in particular a powder or cake, is contained in a single container, especially a sealed container, preferably an injection vial, more preferably a 6 ml injection vial.

According to an embodiment of the first aspect of the present invention, said lyophilized copanlisib-containing solid, in particular a powder or cake, comprises:

copanlisib in a therapeutically effective amount of 68.4 mg, one or more pH adjusters, which is a buffering agent containing a weak acid such as citric acid and a strong base such as sodium hydroxide, and a bulking agent which is mannitol. Said solid substance, in particular a powder or briquette, is hereinafter designated as a lyophilisate.

Said stable lyophilized copanlisib-containing solid, in particular a powder or briquette, contains copanlisib in an amorphous solid state, which shows sufficient stability.

In addition, said stable lyophilized copanlisib-containing solid, particularly a powder or briquette, provides long-term storage as a lyophilized composition, i.e., has a long shelf life and, once reconstituted as a stable reconstituted copanlisib-containing solution, has sufficient time for use after recovery.

Accordingly, said stable lyophilized copanlisib-containing solid, in particular a powder or cake, when reconstituted into a reconstituted solution, is suitable for direct therapeutic use, or for further dilution with a suitable diluent such as sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution, for example, and in turn for further therapeutic use.

To achieve physical stability during and after the lyophilization process, the liquid composition of the invention contains a bulking agent. Bulking agents are commonly used in the art to impart structure and weight to the briquette resulting from lyophilization. Any suitable bulking agent known in the art can be used in connection with the lyophilized composition in accordance with the invention. Suitable bulking agents include, for example, mannitol, dextran, cyclodextrins and glycine, trehalose, sucrose.

The stable, lyophilized, copanlisib-containing solid can then be reconstituted in the following manner.

A method for reconstituting a stable, lyophilized solid containing copanlisib, in particular a powder or briquette, suitable for reconstitution and for therapeutic use, wherein said stable, lyophilized solid, in particular a powder, contains copanlisib, in particular in an amount suitable as a therapeutic dosage, one or more pH regulators, in particular a buffering agent, which in particular contains a weak acid, such as, for example, citric acid, and a strong base, such as, for example, sodium hydroxide, and a bulking agent, said solid, in particularly the powder or cake, has a pH between 4 and 5 (inclusive) when reconstituted with a suitable diluent such as sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution, for example, said reconstitution method includes itself adding to said stable, lyophilized solid, in particular a powder, in a suitable container, especially a sealed container, preferably an injection vial, more preferably a 6 ml injection vial, a diluent such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution;

thus said method provides a stable, reconstituted copanlisib-containing solution suitable for further dilution and for therapeutic use, said reconstituted solution containing copanlisib, in particular in an amount suitable as a therapeutic dose, one or more pH regulators, in particular a buffering agent, which in particular contains a weak acid, such as, for example, citric acid, and a strong base, such as, for example, sodium hydroxide, and a bulking agent, said reconstituted solution has a pH between 4 and 5 (inclusive).

The indicated therapeutic dose is 30, 45, 60 or 80 mg of copanlisib.

In an embodiment, said therapeutic dose is 60 mg of copanlisib.

According to an embodiment, said buffering agent is citric acid and sodium hydroxide.

According to an embodiment, said reconstituted copanlisib-containing solution obtained by said method contains citric acid in an amount of 0.08-15.0% w/v, in particular 0.15-3.0% w/v , in particular 0.15 w/v% of said reconstituted solution.

According to an embodiment, said reconstituted copanlisib-containing solution obtained by said method contains sodium hydroxide in an amount of 0-0.3% w/v, in particular 0.14-0.18% w/v, in in particular, 0.16 w/v% of said reconstituted solution.

According to an embodiment, said reconstituted copanlisib-containing solution obtained by said process contains sodium hydroxide in an amount of 0.01-0.3% w/v, in particular 0.14-0.18% w/v , in particular 0.16 w/v% of said reconstituted solution.

Said bulking agent is mannitol.

According to an embodiment, said reconstituted copanlisib-containing solution obtained by said process contains mannitol in an amount of 1.5-24.0% w/v, in particular 1.5-16.0% w/v, in particular 3.0 w/v% of said reconstituted solution.

According to an embodiment, said diluent is a sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution.

According to an embodiment, said reconstituted copanlisib-containing solution thus obtained contains said copanlisib free base in an amount of 0.5-16.0% w/v, in particular 1.0-8.0% w/v. ./vol.%, in particular 1.5 wt./vol.% of the specified reconstituted solution.

In accordance with an embodiment, in said reconstituted copanlisib-containing solution prepared by said method, said therapeutic dose of copanlisib is present in an amount of 1.5 w/v% of said reconstituted solution, said citric acid is present in an amount of 0.15 w/v .% of said reconstituted solution, said sodium hydroxide is present in an amount of 0.16% w/v of said reconstituted solution, and said mannitol is present in an amount of 3.0% w/v of said reconstituted solution, said diluent is is a 0.9% aqueous solution of sodium chloride, said copanlisib-containing solution having a pH between 4 and 5 (inclusive).

In accordance with an embodiment, said reconstituted copanlisib-containing solution prepared by said method contains copanlisib in an amount of 15 mg/ml.

In an embodiment, said reconstituted copanlisib-containing solution prepared by said method contains 68.4 mg of copanlisib in a volume of 4.55 ml.

In an embodiment, said reconstituted copanlisib-containing solution prepared by said method contains 60 mg of copanlisib in a 4 ml volume.

According to an embodiment, in said reconstituted copanlisib-containing solution prepared by said method:

copanlisib is present in an amount of 68.4 mg, said citric acid is present in an amount of 6.6 mg, said sodium hydroxide is present in an amount of 7.2 mg, and said mannitol is present in an amount of 136.8 mg, said diluent is 0.9% an aqueous solution of sodium chloride, present in an amount of 4.4 ml, said reconstituted copanlisib-containing solution having a pH between 4 and 5 (inclusive), said reconstituted copanlisib-containing solution having a volume of 4.55 ml.

According to an embodiment, said reconstituted copanlisib-containing solution prepared by said method is contained in a single container, especially a sealed container, preferably an injection vial, more preferably a 6 ml injection vial.

Said stable reconstituted copanlisib-containing solution has an acceptable pH and is suitable for direct therapeutic use and/or for further dilution with a suitable diluent, such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, in particular 0.9% aqueous solution sodium chloride, and, in turn, for further therapeutic use.

In an embodiment, said reconstituted copanlisib-containing solution obtained by said method is further diluted with a suitable diluent, such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution.

According to an embodiment, said suitable diluent is a 0.9% aqueous sodium chloride solution, in particular in a volume of 100 ml.

In an embodiment, said reconstituted copanlisib-containing solution prepared by said method is contained in an infusion bag, particularly one made of polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC) or ethylene vinyl acetate (EVA).

This method is hereinafter referred to as a recovery method.

The stable, reconstituted solution containing copanlisib of increased solubility obtained by the said reconstitution method is suitable for further dilution and for therapeutic use, said reconstituted solution contains copanlisib suitable as a therapeutic dose in one or two containers, in particular in one sealed container, one or more pH adjusting agents, a pH buffering agent which contains a weak acid such as citric acid and a strong base such as sodium hydroxide and a bulking agent, said reconstituted solution having been prepared with a suitable solvent such as sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution, e.g.

- 11 044509 said reconstituted solution has a pH between 4 and 5 (inclusive).

The indicated therapeutic dose is 30, 45, 60 or 80 mg of copanlisib.

In an embodiment, said therapeutic dose is 60 mg of copanlisib.

According to an embodiment, said reconstituted copanlisib-containing solution contains citric acid in an amount of 0.08-15.0% w/v, in particular 0.15-3.0% w/v, in particular 0. 15 w/v% of said reconstituted solution.

According to an embodiment, said reconstituted copanlisib-containing solution contains sodium hydroxide in an amount of 0-0.3% w/v, in particular 0.14-0.18% w/v, in particular 0.16% w/v ./vol.% of the specified reconstituted solution.

According to an embodiment of the sixth aspect of the present invention, said reconstituted copanlisib-containing solution contains sodium hydroxide in an amount of 0.01-0.3% w/v, in particular 0.14-0.18% w/v, in particular 0.16 w/v% of said reconstituted solution.

Said bulking agent is mannitol.

According to an embodiment, said reconstituted copanlisib-containing solution contains mannitol in an amount of 1.5-24.0% w/v, in particular 1.5-16.0% w/v, in particular 3.0 w/v% of said reconstituted solution.

According to an embodiment, said diluent is a sterile aqueous sodium chloride solution, in particular a 0.9% aqueous sodium chloride solution.

According to an embodiment, said reconstituted copanlisib-containing solution contains said copanlisib free base in an amount of 0.5-16.0% w/v, in particular 1.0-8.0% w/v, in particular 1.5 w/v% of said reconstituted solution.

In accordance with an embodiment wherein in said reconstituted copanlisib-containing solution, copanlisib is present in an amount of 1.5% w/v of said reconstituted solution, said citric acid is present in an amount of 0.15% w/v of said reconstituted solution. of the reconstituted solution, said sodium hydroxide is present in an amount of 0.16% w/v of said reconstituted solution and said mannitol is present in an amount of 3.0% w/v of said reconstituted solution, said diluent is 0.9% an aqueous solution of sodium chloride, said copanlisib-containing solution has a pH between 4 and 5 (inclusive).

According to an embodiment wherein in said reconstituted copanlisib-containing solution, copanlisib is present in an amount of 68.4 mg, said citric acid is present in an amount of 6.6 mg, said sodium hydroxide is present in an amount of 7.2 mg, and said mannitol is present in an amount of an amount of 136.8 mg, said diluent is a 0.9% aqueous solution of sodium chloride, present in an amount of 4.4 ml, said reconstituted copanlisib-containing solution has a pH between 4 and 5 (inclusive), said reconstituted copanlisib-containing solution has volume 4.55 ml.

In an embodiment, said reconstituted copanlisib-containing solution is contained in a single container, especially a sealed container, preferably an injection vial, more preferably a 6 ml injection vial.

In an embodiment, said reconstituted copanlisib-containing solution contains copanlisib in an amount of 15 mg/ml.

According to an embodiment, said reconstituted copanlisib-containing solution contains 60 mg of copanlisib, especially in a volume of 4 ml.

In an embodiment, said reconstituted copanlisib-containing solution is further diluted with a suitable diluent, such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution.

According to an embodiment, said suitable diluent is a 0.9% aqueous sodium chloride solution, in particular in a volume of 100 ml.

In an embodiment, said reconstituted copanlisib-containing solution is contained in an infusion bag, particularly one made of polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), or ethylene vinyl acetate (EVA).

Said stable reconstituted copanlisib-containing solution contains copanlisib in

- 12 044509 sufficiently high concentrations, i.e. that copanlisib has increased solubility and has an acceptable pH level for its immediate therapeutic use and/or for further dilution with a suitable diluent, such as, for example, sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous chloride solution sodium, and, in turn, for further therapeutic use. This stable reconstituted copanlisib-containing solution provides sufficient time for use after reconstitution.

Said reconstituted solution is hereinafter designated as such.

In accordance with the second aspect of the present invention, the device is a container containing a lyophilized copanlisib-containing solid as described above.

According to an embodiment of the second aspect of the present invention, the device is a sealed container, preferably an injection vial, more preferably a 6 ml injection vial.

experimental part

Example 1. Method for obtaining a stable aqueous bulk solution containing copanlisib, suitable for lyophilization and for therapeutic use

This method is hereinafter designated as a manufacturing method, and the bulk solution obtained in this way is hereinafter designated as a bulk solution.

The method for preparing bulk copanlisib solution is as follows:

Water for injection is loaded into the production vessel. Copanlisib is transferred into a vessel and dissolved with stirring at pH 2-3. A bulking agent and a buffering agent are added and dissolved with stirring. The pH of the solution is adjusted to pH 5.0 using an appropriate amount of sodium hydroxide solution. After this, sterile filtration of the bulk solution is carried out and bottles are filled with the solution.

Solubility:

Table 1. Solubility of copanlisib dihydrochloride

Solvent Solubility at 25 °C [mg/ml] Descriptive term ^a Water 2.3 10 ² easily soluble 0.1 M hydrochloric acid 1.9 10 ² easily soluble Buffer (phosphate) pH 1 5.1 10 ² easily soluble Buffer (phosphate) pH 2 2.5 10 ² easily soluble Buffer (citrate) pH 3 2.5 10 ² easily soluble Buffer (citrate) pH 4 2.5 Yu ¹ soluble Buffer (citrate) pH 5 6.2 · 10' ¹ very slightly soluble Buffer (phosphate) pH 6 1 10' ⁵ practically insoluble Buffer (phosphate) pH 7 < 1 10' ⁵ practically insoluble and in accordance with the European Pharmacopoeia/US Pharmacopoeia

Chemical stability in solution

Various aqueous solutions (0.05% copanlisib; after addition of 50% 2-propanol, [the buffer solution was filtered through a 0.5 μm membrane filter]) were stored at 25°C and 70°C for 24 hours and one week. An HPLC analytical method was developed to determine samples and organic contaminants of copanlisib, as well as degradation products.

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Table 2. Copanlisib hydrolytic stability results

Conditions Appearance Sum Max, separate products products decomposition decomposition [area-%] [area-%] Water: Original slightly colored solution 2.79 0.25 24 hours, 25°C slightly colored solution 3.43 0.23 24 hours, 70°C slightly colored solution 58.00 25.89 1 week, 25°C slightly colored solution 5.33 0.54 1 week, 70°C slightly colored solution 98.59 45.44 Buffer pH 7: Original slightly colored cloudy solution 3.15 0.23 24 hours, 25°C slightly colored cloudy solution 3.22 0.20 24 hours, 70°C slightly colored solution 56.06 23.25 1 Week, slightly colored cloudy 4.85 0.82 25°С solution 1 week, 70°C slightly colored solution 97.65 39.01 0.1 M HC1: Initial slightly colored solution 5.87 1.13 24 hours, 25°С slightly colored solution 8.75 1.90 24 hours, 70°C slightly colored solution 92.49 22.82 1 week, 25°C slightly colored solution 24.27 7.15 1 week, 70°C slightly colored solution 100.00 25.48 0.1 M NaOH: Original slightly colored solution 30.72 6.51 24 hours, 25°С slightly colored solution 45.40 10.02 24 hours, 70°C slightly colored solution 99.88 23.94 1 week, 25°C slightly colored solution 86.64 22.03 1 week, 70°C slightly colored solution 99.90 32.63

Physical Stability

With regard to the required holding time for the bulk solution during the manufacturing process, including vial filling and freezing of the solution, chemical and physical stability in the solution is of fundamental importance even for the development of a lyophilisate.

Reconstituted solutions must also demonstrate sufficient physical and chemical stability (stability in use) to avoid harm upon final administration to the patient.

The chemical stability of copanlisib bulk solution increases with increasing pH. In particular, the formation of the major degradation product aminoethyltrioxoquinazolinepyramide (hereafter abbreviated as AETQP) revealed a clear pH dependence in the studied range from pH 3 to pH 6, demonstrating higher stability at higher pH.

However, at pH 6, precipitation of bulk copanlisib dihydrochloride solution was detected visually under stress conditions. Therefore, the pH of the bulk solution should be below pH 6.

In fig. 1/4 shows the pH-dependent formation of AETQP in bulk solution of copanlisib at 2-8°C, 25°C and 40°C.

In fig. 2/4 shows the pH-dependent stability of bulk solution of copanlisib.

Bulk solution (40 mg/mL, copanlisib free base)

Studies regarding the physical stability of copanlisib bulk solution were performed after filling the vials. Visual inspection was performed on bottles filled with bulk solution approximately 17 days after production and storage at 28°C without any results. Further studies were performed later, approximately 3 weeks after production of the bulk solution (Table 3). No precipitation was observed during the test period of an additional 14 days at room temperature or when stored at 2-8°C. In summary, bulk solution of copanlisib prepared under the described conditions, including sterile filtration and filling of glass vials, did not tend to precipitate within 5 weeks of manufacture.

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Table 3. Effect of temperature on the physical stability of bulk solution in vials (n=3), studies began ca. 3 weeks after production and storage at 2-8°C (holding time for bulk solution)

Observations 20°С 4°C Start (3 weeks after production) clear solution clear solution 24 hours clear solution clear solution 48 hours clear solution clear solution 5 days clear solution clear solution 7 days clear solution clear solution 14 days clear solution clear solution

Example 2. Method for lyophilization of a stable aqueous bulk solution containing copanlisib, suitable for lyophilization and for therapeutic use

Said method is hereinafter designated as a lyophilization method, the lyophilized solid, in particular the powder or cake thus obtained, is hereinafter designated as lyophilisate and the stable lyophilized copanlisib-containing solid, in particular the powder or cake, suitable for therapeutic use, containing, inter alia , a therapeutic dose of copanlisib in one or two containers, especially in one sealed container, is hereinafter referred to as a ready-to-reconstitute lyophilisate or drug product.

Lyophilization method

Lyophilization or freeze-drying is a well-known process for stabilizing liquid formulations. A typical lyophilization process can be divided into three stages: freezing including annealing, primary drying and secondary drying.

The initial freezing of the liquid composition causes the water to turn into ice and the solutes to crystallize. An additional annealing step ensures complete crystallization of the dissolved substances. During primary drying, frozen ice is sublimated under vacuum. Residual moisture is evaporated from the matrix during secondary drying. Finally, a dry and solid briquette is obtained containing the dissolved substances of the original solution.

The lyophilization method was developed on a laboratory scale (HOF, serial no.: room 41239). For the production of lyophilisate on a pilot and industrial scale HOF, factory. No.: com. so far we have used 41041.

Table 4a. Nominal loading and filling, including overfill and quantity of copanlisib free base and copanlisib dihydrochloride for the 60 mg dose

Nominal filling Filling, including overflow Copanlisib free base 60 mg 68.4 mg Copanlisib dihydrochloride 69.1 mg 78.8 mg Bulk solution volume/bottle 1.5 ml 1.71 ml Recovery volume solution/bottle 4.0 ml 4.55 ml

The freeze drying parameters are as follows:

Table 4b. Comparison of freeze-drying processes applied to copanlisib lyophilisates

Freeze-drying process applied to copanlisib 80 mg lyophilisate Initial freeze-drying process applied to copanlisib 60 mg lyophilisate Final freeze-drying process applied to copanlisib 60 mg lyophilisate Filling bulk solution Nominal fill 2.0ml plus additional 0.1ml overflow Nominal fill 1.5ml plus additional 0.07ml or 0.21ml overflow Nominal fill 1.5ml plus additional 0.21ml overflow

- 15 044509

Freezing Ramp to -45°C, hold for 2 hours. Ramp to -45°C, hold for 1.5 hours. Ramp to -45°C, hold for 1.5 hours. Annealing -20 °C for 4 hours. -20 °C for 2 hours. -20 °C for 3.5 hours. Primary drying Isothermal retention at -5 °C /100 µbar for 23 hours. Drying ramp from -45 °C to +30 °C from 0.08 °C/min / 100 µbar, duration 15.6 hours. Isothermal retention at +5 °C /110 µbar for 14 hours. Secondary drying Ramp up to +40 °C, hold for 8 hours. Ramp up to +35 °C, hold for 6.2 hours. Ramp up to +35 °C, hold for 4 hours.

Table 5. Final process including ranges

Stage Temperature Pressure Time Freezing from -42 to -48 °C atmospheric > 1.5 hours Annealing -18 to -24 °C atmospheric > 3.0 hours Primary drying 1-9 °C 70-150 µbar > 10 hours Secondary drying 30-40 °C 70-150 µbar 3-8 hours Storage until unloading 5-25 °C 70-150 µbar < 72 h.

Determination of solubility

The solubility of copanlisib dihydrochloride was determined using the flask method by stirring copanlisib in the solvents to be tested in a thermostatically controlled water bath at 25°C for 19 hours. The suspension was centrifuged before HPLC analysis. The concentration was monitored by HPLC against an external standard.

Determination of pH The pH of copanlisib solutions is determined potentiometrically in accordance with Eur. Pharmacopoeia.

Storage stability experiment

During the development of lyophilisates of copanlisib, analysis of the physicochemical state of the lyophilized product confirmed only crystalline peaks based on mannitol (present in modification III) and did not reveal evidence of crystallization of the drug substance. Stress testing of technical and clinical batches has shown that this behavior persists for at least 18 months of storage at 30°C/75% RH.

Table 6. Solid State of 60 mg Copanlisib Injection Lyophilisate after Stress Testing

Batch number Storage conditions Solid state (sRSA) Batch A 18 months, 30 °C/75% RH Mannitol mod. III, amorphous drug substance Party B 18 months, 30 °C/75% RH Mannitol mod. III, amorphous drug substance Party C 12 months, 30 °C/75% RH Mannitol mod. III, amorphous drug substance

Lots A, B and C have the following quantities of components, as shown in the table below. 6a. Table 6a. Amounts of components in Lots A, B and C of copanlisib 60 mg lyophilisate

Lyophilisate (w/w%) Copanlisib (free base) 31 Mannitol 62 Lemon acid 3

Sodium hydroxide, which is necessary to adjust the pH, is contained in the required amount.

There are also batches of 80 mg drug products that were stored at the intended storage conditions of 2-8°C, as well as at elevated conditions of 25°C/60% RH, in which it was confirmed that the amorphous state of the drug substance was maintained even for 4 years of storage (Table 7).

- 16 044509

Table 7. Solid state of copanlisib 80 mg injection lyophilisate after long-term testing

Batch number Storage conditions Solid state (sRSA) Party D 48 months, 2-8 °C Mannitol mod. III, amorphous drug substance Party D 48 months, 25 °C/60% RH Mannitol mod. III, amorphous drug substance Party E 37 months, 2-8 °C Mannitol mod. III, amorphous drug substance Party E 37 months, 25 °C/60% RH Mannitol mod. III, amorphous drug substance

Thus, it can be confirmed that the amorphous state of the drug substance, which ensures rapid dissolution upon reconstitution, is maintained throughout the entire shelf life of the product.

In addition, the residual moisture content of the lyophilisate was monitored during long-term stability studies. The results for the 60 mg and 80 mg lyophilisate formulations are presented in the following tables. With respect to all tested batches and conditions, the water content in the lyophilisates was within the specified limit of 2.0%. Solid state studies (SSA) of the lyophilisate confirmed the amorphous state of the drug substance, regardless of the water content tested. Moreover, mannitol is still present in modification III.

The figure shows a 3/4 X-ray powder diffraction pattern of copanlisib 60 mg injection after 18 months at 30°C and 75% relative humidity (Batch A, see Table 6a).

In table Figure 7a shows the corresponding 2Θ values of the diffraction peaks.

For reference, the X-ray powder diffraction pattern of modification III of D-mannitol (also known as the δ form) is shown in FIG. 4/4. The diffraction pattern was modeled based on the single-crystal X-ray structure published by Botez et al. 2003 (see C.E. Botez, P.W. Stephens, S. Nunes, R. Suryanarayanan, Powder Diffr. (2003), 18, 214).

Table 7a. 2θ diffraction peak values of copanlisib 60 mg injection after 18 months of storage at 30°C and 75% relative humidity (Batch A, see Table 6a)

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Table 8. Residual moisture results of copanlisib 60 mg lyophilisate after 12 months

Batch number Storage conditions Residual moisture [%] Specification max. 2.0% Initial 12 months Batch A 30 °C/75% RH 0.6 0.9 2-8°С 0.5 Party B 30 °C/75% RH 0.6 0.9 25 °C/75% RH 0.8 2-8°С 0.6 Part F 25 °C/60% RH 0.5 0.8

Table 9. Residual moisture results of copanlisib 80 mg lyophilisate after 36 months

Batch number Storage conditions Residual moisture [%] Specification max. 2.0% Initial 36 months Party G 25 °C/60% RH 0.7 1.5 2-8 °C 1.0 Party D 25 °C/75% RH 1.1 1.7 2-8 °C 1.1 Party E 25 °C/60% RH 1.0 But. 2-8 °C 0.9

Since no change was observed in the amorphous state of the drug substance in the lyophilisate, the following moisture stress test was performed with copanlisib 60 mg injection lyophilisate (Table 10). The cap and stopper of copanlisib lyophilisate vials were removed and the vials were stored at 25°C/60% RH for up to 48 hours to increase the water content of the lyophilisate. Residual moisture was determined by Karl-Fischer titration. An equilibrium of 3.5% moisture is achieved after 24 hours of open storage.

However, no changes in the amorphous phase of the composition were detected by pXRD up to 3.5% residual moisture and 48 h of open storage, demonstrating a very low tendency for the amorphous phase to crystallize.

Table 10. Effect of residual moisture on the solid state of copanlisib 60 mg injection lyophilisate (Lot K)

Open storage in climate chamber [h] Residual water after open storage [%] Solid state (sRSA) 0 0.4 Mannitol mod. III, amorphous drug substance 0.5 0.8 Mannitol mod. III, amorphous drug substance 1 1.4 Mannitol mod. III, amorphous drug substance 1.5 1.6 Mannitol mod. III, amorphous drug substance 2 1.7 Mannitol mod. III, amorphous drug substance 3 2.4 Mannitol mod. III, amorphous drug substance 4 h,o Mannitol mod. III, amorphous drug substance 24 3.6 Mannitol mod. III, amorphous drug substance 48 3.5 Mannitol mod. III, amorphous drug substance

Example 3: Method for reconstituting a stable, lyophilized, copanlisib-containing solid, in particular a powder or briquette, suitable for reconstitution and therapeutic use.

This method is hereinafter designated as a reconstitution method, and the stable, reconstituted copanlisib-containing solution of increased solubility obtained in this manner, suitable for further dilution and for therapeutic use, is hereinafter designated as a reconstituted solution.

Reconstituted (15 mg/mL, copanlisib free base) and diluted solution (0.56 mg/mL, copanlisib free base)

Lyophilisates prepared in the laboratory were reconstituted without removing the stopper with an aqueous solution of sodium chloride 0.9% to obtain a solution of 15 mg/ml copanlisib free base.

Based on the results of the bulk solution of copanlisib, the effect of temperature was also studied for the reconstituted solution (Table 11).

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Table 11. Effect of temperature on physical stability of reconstituted solution (15 mg/mL copanlisib free base), Lot H (stability when reconstituted solution is used)

Observations 20°C bottle No. I 4°C bottle No. III Start clear solution clear solution 1 day clear solution clear solution 2 days clear solution clear solution 5 days clear solution clear solution 6 days clear solution clear solution 7 days clear solution clear solution

Example 4: Copanlisib in Patients with Relapsed or Refractory Indolent B Cell Lymphoma

Methods:

Eligible patients were patients with indolent B-cell non-Hodgkin lymphoma (4 subtypes: follicular (FL), marginal zone (LMZ), small cell lymphocytic (SLL) and lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (WPL-MB)) and relapse after, or refractory to > 2 previous lines of treatment. Previous treatment should have included rituximab and an alkylating agent. Copanlisib (60 mg, i.v.) was administered periodically on days 1, 8, and 15 of a 28-day cycle. The primary endpoint was objective tumor response rate (OOP) assessed by independent radiological review (Cheson et al., JCO 20:579, 2007).

Results:

The full analysis population included 142 patients, of whom 141 patients had indolent lymphoma (FL/LMZ/MLL/LPL-MB: 104/23/8/6). At the time of the primary analysis, the median duration of treatment was 22 weeks (range 1–105); 46 patients remained under treatment. The most common treatment-related adverse events (AEs) (all grades/grade 3+) were transient hyperglycemia (49%/40%) and hypertension (29%/23%). Other AEs of interest included neutropenia (25%/19%), diarrhea (18%/4%), pulmonary infection (14%/11%), pneumonitis (7%/1.4%), and colitis (0. 7%/0.7%). No colonic perforation was observed. There were two non-fatal opportunistic infections. Laboratory toxicities of interest were primarily grade 1, including alanine aminotransferase (23% all grade/19% grade 1) and aspartate aminotransferase (28%/25%). There were 6 deaths, 3 of which were associated with copanlisib: pulmonary infection, respiratory failure, and thromboembolic complications.

The overall response rate (OOP) was 59.2%, including 12.0% complete response (CR) and 47.2% partial response (PR), with stable disease in 29.6% of patients and progressive disease in 2.1% of patients. In the FL subpopulation, OOP was 58.7%, including 14.4% CR and 44.2% PR. In the LMZ subpopulation, the OOP was 69.6%, including 8.7% CR and 60.9% PR. The estimated median duration of Kaplan-Meier (KM) response in the full analysis population was 687 days (range 0687) and 370 days (range 0-687) in the FL subpopulation. The KM calculation of median PFS was 340 days (range 0–736). Median overall survival has not yet been reached.

Conclusions:

Treatment of patients with relapsed or refractory indolent B-cell lymphoma with copanlisib resulted in durable tumor responses. Copanlisib administration had a manageable safety profile, with low rates of severe hepatic enzymopathy, diarrhea, or inflammatory events, and low rates of opportunistic infections, fatal infections, or other fatal serious adverse events.

A stable, reconstituted copanlisib-containing solution of increased solubility, suitable for further dilution and for therapeutic use, said reconstituted solution contains a therapeutic dose of copanlisib in one or two containers, in particular in one sealed container, one or more pH-buffering agent, which contains a weak acid such as citric acid and a strong base such as sodium hydroxide, and a bulking agent, said reconstituted solution has been prepared with a suitable solvent such as sterile water for injection or sterile aqueous sodium chloride, especially 0.9 % aqueous solution of sodium chloride, for example said reconstituted solution has a pH between 4 and 5 (inclusive), can be used for the treatment or prevention of cancer, especially non-Hodgkin's lymphoma (NHL), such as 1st line, 2nd line or before 9 previous treatments, relapsed, refractory, indolent NHL, in particular, follicular lymphoma (FL), marginal zone lymphoma (LMZ), small cell lymphocytic lymphoma (SCL),

- 19 044509 lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (LPL-MV) or aggressive NHL, in particular diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), peripheral T-cell lymphoma (PTCL), transformed lymphoma (TL) ) or chronic lymphocytic leukemia (CLL) as monotherapy or in combination with one or more immunotherapy, chemotherapy or immunochemotherapeutic anticancer agents.

In accordance with an embodiment, said copanlisib-containing solution is defined in any of the aspects described above.

A stable, reconstituted copanlisib-containing solution of increased solubility, suitable for further dilution and for therapeutic use, said reconstituted solution contains a therapeutic dose of copanlisib in one or two containers, in particular in one sealed container, one or more pH regulators, a buffering agent, which contains a weak acid such as citric acid and a strong base such as sodium hydroxide and a bulking agent, said reconstituted solution has been prepared with a suitable solvent such as sterile water for injection or sterile aqueous sodium chloride solution, especially 0 A .9% aqueous solution of sodium chloride, for example, said reconstituted solution having a pH between 4 and 5 (inclusive), can be used for the treatment or prevention of cancer, especially non-Hodgkin's lymphoma (NHL), such as 1st line, 2nd line or up to 9 prior treatments, relapsed, refractory, indolent NHL, in particular follicular lymphoma (FL), marginal zone lymphoma (Marginal zone lymphoma), small cell lymphocytic lymphoma (SLL), lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (WPL-MB) or aggressive NHL, in particular diffuse B large cell lymphoma (DLCL), mantle cell lymphoma (MCL), peripheral T cell lymphoma (PTCL), transformed lymphoma (TL) or chronic lymphocytic leukemia (CLL), as monotherapy or in combination with one or more immunotherapies, chemotherapeutic or immuno-chemotherapeutic anticancer agents.

A stable, reconstituted, copanlisib-containing, solution of increased solubility suitable for further dilution and for therapeutic use, said reconstituted solution contains a therapeutic dose of copanlisib in one or two containers, in particular in one sealed container, one or more pH-buffering agent, which contains a weak acid such as citric acid and a strong base such as sodium hydroxide, and a bulking agent, said reconstituted solution was prepared with a suitable solvent such as sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous solution sodium chloride, for example said reconstituted solution has a pH between 4 and 5 (inclusive), for the preparation of a medicament for the treatment or prevention of cancer, especially non-Hodgkin's lymphoma (NHL), such as 1st line, 2nd line or up to 9 previous treatment, relapsed, refractory, indolent NHL, particularly follicular lymphoma (FL), marginal zone lymphoma (Marginal zone lymphoma), small cell lymphocytic lymphoma (SLL), lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (LPL-W), or aggressive NHL, particularly diffuse B - large cell lymphoma (DLCL), mantle cell lymphoma (MCL), peripheral T-cell lymphoma (PTCL), transformed lymphoma (TL) or chronic lymphocytic leukemia (CLL), as monotherapy or in combination with one or more immunotherapeutic agents, chemotherapeutic or immuno-chemotherapeutic anticancer agents.

The solution may be used in a method of treating or preventing cancer, especially non-Hodgkin's lymphoma (NHL), such as 1st line, 2nd line, or up to 9 prior treatments, relapsed, refractory, indolent NHL, in particular follicular lymphoma ( FL), marginal zone lymphoma (LMZ), small cell lymphocytic lymphoma (SLL), lymphoplasmacytic lymphoma/Waldenström macroglobulinemia (LPL-W) or aggressive NHL, in particular diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL) ), peripheral T-cell lymphoma (PTCL), transformed lymphoma (TL), or chronic lymphocytic leukemia (CLL) in a subject, comprising administering to said subject a stable, reconstituted copanlisib-containing solution of increased solubility suitable for further dilution and therapeutic use , wherein said reduced solution contains

- 20 044509 a therapeutic dose of copanlisib in one or two containers, in particular in one sealed container, one or more pH-buffering agent which contains a weak acid such as citric acid and a strong base such as sodium hydroxide and a bulking agent agent, said reconstituted solution has been prepared with a suitable solvent such as sterile water for injection or sterile aqueous sodium chloride solution, especially 0.9% aqueous sodium chloride solution, for example said reconstituted solution has a pH between 4 and 5 (inclusive), in as monotherapy or in combination with one or more immunotherapeutic, chemotherapeutic or immunochemotherapeutic anticancer agents.

Said copanlisib-containing solution is defined in any of the aspects described above.

Said copanlisib-containing solution contains 30, 45, 60 or 80 mg of copanlisib.

In an embodiment, said copanlisib-containing solution contains 60 mg of copanlisib.

In an embodiment, said administration of said copanlisib-containing solution is administered by administering a fixed dose of 60 mg of copanlisib to a patient as a 1-hour intravenous infusion on days 1, 8, and 15 of a 28-day treatment cycle on an interval schedule (three weeks of administration and one week break).

In accordance with an embodiment, said copanlisib-containing solution is administered in combination with a chemotherapeutic anticancer agent, in particular a chemotherapeutic anticancer agent selected from the group consisting of

131I-chTNT, abarelix, abiraterone, aclarubicin, adalimumab, ado-trastuzumab emtansine, afatinib, aflibercept, aldesleukin, alectinib, alemtuzumab, alendronic acid, alitretinoin, altretamine, amifostine, aminoglutethimide, hexylaminolevulinate, amrub icin, amsacrine, anastrozole, ancestim, anethole dithioletion , anetumab ravtansin, angiotensin II, antithrombin III, aprepitant, arcitumomab, arglabin, arsenic trioxide, asparaginase, atezolizumab, axitinib, azacitidine, basiliximab, belotecan, bendamustine, besilezomab, belinostat, bevacizumab, bexarotene, bicalutamide, bisantrene, bleomycin, blinatumomab, bortezomib , buserelin, bosutinib, brentuximab vedotin, busulfan, cabazitaxel, cabozantinib, calcitonin, calcium folinate, calcium levofolinate, capecitabine, capromab, carbamazepine, carboplatin, carboquone, carfilzomib, carmofur, carmustine, catumaxomab, celecoxib, celmoleuquin, ceritinib, tse tuximab, chlorambucil, chlormadinone, chlormethine, cidofovir, cinacalcet, cisplatin, cladribine, clodronic acid, clofarabine, cobimetinib, crizantaspase, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daratumumab, darbepoetin alfa, dabrafenib, dasatinib, daunorubicin , decitabine, degarelix, denileukin diftitox , denosumab, depreotide, deslorelin, dianhydrogalactitol, dexrazoxane, dibrospidium chloride, dianhydrogalactitol, diclofenac, dinutuximab, docetaxel, dolasetron, doxifluridine, doxorubicin, doxorubicin + estrone, dronabinol, eculizumab, edrecolomab, eliptinium acetate, elotuzum b, eltrombopag, endostatin, enocitabine, enzalutamide , epirubicin, epithiostanol, epoetin alfa, epoetin beta, epoetin zeta, eptaplatin, eribulin, erlotinib, esomeprazole, estradiol, estramustine, ethinyl estradiol, etoposide, everolimus, exemestane, fadrozole, fentanyl, filgrastim, fluoxymesterone, floxuridine, fludarabine, fluorouracil, flutamide, folinic acid, formestane, fosaprepitant, fotemustine, fulvestrant, gadobutrol, gadoteridol, gadoteric acid meglumine, gadoversetamide, gadoxetic acid, gallium nitrate, ganirelix, gefitinib, gemcitabine, gemtuzumab, glucarpidase, glutoxim, GM-CSF, goserelin, granisetron, granulocyte colony-stimulating factor , histamine dihydrochloride, histrelin, hydroxyurea, I-125 grains, lansoprazole, ibandronic acid, ibritumomab tiuxetan, ibrutinib, idarubicin, ifosfamide, imatinib, imiquimod, improsulfan, indysetron, incadronic acid, mebulate ingenol, interferon alpha, interferon beta, interferon gamma, iobitridol, iobenguane (123I), iomeprol, ipilimumab, irinotecan, itraconazole, ixabepilone, ixazomib, lanreotide, lansoprazole, lapatinib, lazocholine, lenalidomide, lenvatinib, lenograstim, lentinan, letrozole, leuprorelin, levamisole, levonorgestrel, Levot iroxine sodium, lisuride, lobaplatin, lomustine, lonidamine, masoprocol, medroxyprogesterone, megestrol, melarsoprol, melphalan, mepithiostane, mercaptopurine, mesna, methadone, methotrexate, methoxalen, methylaminolevulinate, methylprednisolone, methyltestosterone, metyrosine, mifamurtide, miltefosine, miriplatin, mitobronitol , mitoguazone, mitolactol, mitomycin, mitotane, mitoxantrone, mogamulizumab, molgramostim, mopidamol, morphine hydrochloride, morphine sulfate, nabilone, nabiximols, nafarelin, naloxone + pentazocine, naltrexone, nartograstim, necitumumab, nedaplatin, nelarabine, neridronic acid, netupitant/palonosetron, nivolumab, pentetreotide, nor lotinib, nilutamide, nimorazole , nimotuzumab, nimustine, nintedanib, nitracrine, nivolumab, obinutuzumab, octreotide, ofatumumab, olaparib, olaratumab, omacetaxine mepesuccinate, omeprazole, ondansetron, oprelvekin, orgotein, orilotimod, osimertinib, oxaliplatin, oxycodone, oxymetholone, ozogamicin, p53 gene therapy, paclitaxel, palbociclib, palifermin, palladium-103 grains, palonosetron, pamidronic acid, panitumumab, panobinostat, pantoprazole, pazopanib, pegaspargase, PEG-epoetin beta (methoxy PEG-epoetin

- 21 044509 beta), pembrolizumab, pegfilgrastim, peginterferon alfa-2b, pembrolizumab, pemetrexed, pentazocine, pentostatin, peplomycin, perflubutan, perfosfamide, pertuzumab, picibanil, pilocarpine, pirarubicin, pixantrone, plerixafor, plicamycin, polyglusam, polyestradiol phosphate, polyvinylpyrrolidone + sodium hyaluronate, polysaccharide-K, pomalidomide, ponatinib, sodium porfimer, pralatrexate, prednimustine, prednisone, procarbazine, procodazole, propranolol, quinagolide, rabeprazole, racotumumab, radium-223 chloride, radotinib, raloxifene, raltitrexed, ramosetron, ramucirumab, ranimu Steen, Rasburikas , razoxane, refametinib, regorafenib, risedronic acid, rhenium-186 etidronate, rituximab, rolapitant, romidepsin, romiplostim, romurtide, rucaparib, samarium (153Sm) lexidronam, sargramostim, satumomab, secretin, siltuximab, sipuleucel-T, sisofi ran, sobuzoxan, glycididazole sodium, sonidegib, sorafenib, stanozolol, streptozocin, sunitinib, talaporfin, talimogene laherparepvec, tamibarotene, tamoxifen, tapentadol, tazonermine, tetseleukin, technetium (99 mTc) nofetumomab merpentane, 99mTc-HYNIC-[Tyr3]-octreotide, tegafur, tegafur + gimeracil + oteracil, temoporfin, temozolomide, temsirolimus, teniposide, testosterone, tetrofosmin, thalidomide, thiotepa, thymalfasin, thyrotropin alfa, thioguanine, tocilizumab, topotecan, toremifene, tositumomab, trabectedin, trametinib, tramadol, trastuzumab, trastuzumab emtansine, threo sulfan, tretinoin, trifluridine + tipiracil, trilostane, triptorelin, trametinib, trofosfamide, thrombopoietin, tryptophan, ubenimex, valatinib, valrubicin, vandetanib, vapreotide, vemurafenib, vinblastine, vincristine, vindesine, vinflunine, vinorelbine, vismodegib, vorinostat, vorozol, ytrium-9 glass microbeads 0, zinostatin , zinostatin stimalamer, zoledronic acid, zorubicin.

The above administration is carried out orally or parenterally. For copanlisib, parenteral routes of administration are preferred. Methods of parenteral delivery include topical, intra-arterial (directly into the tumor), intramuscular, subcutaneous, intramedullary, intrathecal, intraventricular, intravenous, intraperitoneal or intranasal administration. In addition to the active ingredients, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers containing excipients and excipients that facilitate processing of the active compounds into preparations that can be used pharmaceutically. More detailed information on methods of preparation and administration can be found in the latest edition of Remington's Pharmaceutical Sciences (Published by Maack Publishing Co, Easton, Pa.).

The compositions in accordance with the invention can be administered using an injector, pump, syringe or any other devices/infusion devices known in the art, as well as under the influence of gravity. A needle or catheter can be used to introduce the compositions of the present invention into the patient through certain parenteral routes.

Pharmaceutical compositions suitable for use include compositions in which the active ingredients are contained in an effective amount to achieve the intended purpose, that is, to treat a particular disease. Determination of the effective dose is within the competence of those skilled in the art.

For copanlisib, the therapeutically effective dose can be initially assessed either in cell culture assays, such as tumor cells, or in animal models, typically mice, rabbits, dogs, pigs or monkeys. An animal model is also used to achieve the desired concentration range and route of administration. Such information can then be used to determine acceptable doses and routes of administration to humans.

Copanlisib-containing compositions are formulated to be acceptable for therapeutic use. The term therapeutic use refers to treatment comprising administering to a subject in need of treatment a therapeutically effective amount of copanlisib. The concept of a therapeutically effective amount is defined as an amount of copanlisib that is sufficient to cause tumor cell death by apoptosis and inhibition of proliferation of the primary malignant B cell lineage, either as a single dose or in accordance with a multiple-dose regimen, separately or in combination with other agents, which results in relief of the unfavorable condition, but is still in an amount that is toxicologically acceptable. The subject may be a human or a non-human animal (eg, a rabbit, rat, mouse, dog, monkey, or other lower-order primate).

The exact dosage can be selected by the individual physician based on the patient being treated. Dosage and administration are adjusted to provide sufficient levels of the active ingredient or to maintain the desired effect. Additional factors that may be taken into account include the severity of the disease state, such as the size and location of the tumor; age, weight and gender of the patient; diet, timing and frequency of dosing, drug combination(s), sensitivity to reaction, and tolerance/response to therapy.

Long-acting pharmaceutical compositions can be administered, for example, every 3-4 days, once a week, once every two weeks, or once every three weeks, depending on the half-life and elimination rate of the particular composition.

- 22 044509

In accordance with the present invention, copanlisib-containing compositions can be administered to a patient as a fixed dose of 30, 45, 60, or 80 mg of copanlisib as a 1-hour intravenous infusion on days 1, 8, and 15 of a 28-day treatment cycle on an interval schedule (three weeks of introduction and one week break).

In accordance with the present invention, copanlisib-containing compositions are administered to a patient as a fixed dose of 60 mg of copanlisib as a 1-hour intravenous infusion on days 1, 8, and 15 of a 28-day treatment cycle on an interval schedule (three weeks on and one week off).

According to a third aspect, the present invention includes

Item 1. A therapeutic, lyophilized, copanlisib-containing solid containing copanlisib, in a therapeutically effective amount, one or more pH adjusters, a buffering agent containing a weak acid such as citric acid and a strong base such as sodium hydroxide, and a bulking agent which is mannitol, wherein the indicated therapeutically effective amount is 30, 45, 60 or 80 mg of copanlisib.

Claim 2. Lyophilized copanlisib-containing solid according to claim 1, which contains citric acid in an amount of 0.7-75% w/w, in particular 1.5-30% w/w, in particular 2, 98% w/w of said lyophilized copanlisib-containing solid.

Clause 3. Lyophilized copanlisib-containing solid according to claim 1 or 2, which contains sodium hydroxide in an amount of 0.01-6 w/w%, in particular 3.3-4.0 w/w%, in particular 3.7% w/w of said lyophilized copanlisib-containing solid.

Claim 4. Lyophilized copanlisib-containing solid according to any one of claims 1 to 3, which contains mannitol in an amount of 15.4-82.1% w/w, in particular 30.8-82.1 w/w .%, in particular 61.6 w/w%, of said lyophilized copanlisib-containing solid.

Clause 5. The lyophilized copanlisib-containing solid according to any one of claims. 1-4, which contains copanlisib in an amount of 7.7-92.4 wt./wt.%, in particular 7.7-61.6 wt./wt.%, in particular 30.8 wt./wt. % of lyophilized copanlisib-containing solid.

Claim 6. The lyophilized copanlisib-containing solid according to any one of claims 1 to 5, wherein copanlisib is present in an amount of 30.8 w/w% of said lyophilized copanlisib-containing solid, said citric acid is present in an amount of 2, 98 w/w% of said lyophilized copanlisib-containing solid, said sodium hydroxide is present in an amount of 3.7 w/w% of said lyophilized copanlisib-containing solid, and said mannitol is present in an amount of 61.6 wt ./wt.%, from the specified lyophilized solid containing copanlisib.

Claim 7. The lyophilized copanlisib-containing solid according to any one of claims 1 to 6, wherein copanlisib is present in an amount of 30, 45 or 60 mg, said citric acid is present in an amount of 5.8 mg, said sodium hydroxide is present in an amount of 6, 3 mg and the said mannitol is present in an amount of 120 mg.

Clause 8. The lyophilized copanlisib-containing solid according to any one of claims. 1-6, in which:

copanlisib is present in an amount of 80 mg, said citric acid is present in an amount of 7.7 mg, said sodium hydroxide is present in an amount of 8.4 mg, and said mannitol is present in an amount of 160 mg.

Item 9. A therapeutic, lyophilized, copanlisib-containing solid containing copanlisib in a therapeutically effective amount of 68.4 mg, one or more pH adjusters, a buffering agent containing a weak acid such as citric acid and a strong base such as hydroxide sodium, and a bulking agent, which is mannitol. Clause 10. Lyophilized copanlisib-containing solid according to any one of claims. 1-9, which is a powder or briquette.

Clause 11. A device which is a container containing a lyophilized copanlisib-containing solid according to any one of claims 1 to 10.

Clause 12. The device of claim 11, which is a sealed container, preferably an injection vial, more preferably a 6 ml injection vial.

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Claims (12)

CLAIM 1. A therapeutic, lyophilized, copanlisib-containing solid containing copanlisib in a therapeutically effective amount, one or more pH adjusters, a buffering agent containing a weak acid such as citric acid and a strong base such as sodium hydroxide, and a bulking agent , which is mannitol in which the indicated therapeutically effective amount is 30, 45, 60 or 80 mg of copanlisib. 2. Lyophilized copanlisib-containing solid according to claim 1, which contains citric acid in an amount of 0.7-75% w/w, in particular 1.5-30% w/w, in particular 2.98 w/w% of said lyophilized copanlisib-containing solid. 3. Lyophilized copanlisib-containing solid according to claim 1 or 2, which contains sodium hydroxide in an amount of 0.01-6 wt./wt.%, in particular 3.3-4.0 wt./wt.%, in specifically 3.7% w/w of said lyophilized copanlisib-containing solid. 4. Lyophilized copanlisib-containing solid according to any one of claims 1 to 3, which contains mannitol in an amount of 15.4-82.1 wt./wt.%, in particular 30.8-82.1 wt./wt. %, in particular 61.6 w/w% of said lyophilized copanlisib-containing solid. 5. Lyophilized copanlisib-containing solid according to any one of claims 1 to 4, which contains copanlisib in an amount of 7.7-92.4 wt./wt.%, in particular 7.7-61.6 wt./wt. %, in particular 30.8 w/w% of the lyophilized copanlisib-containing solid. 6. The lyophilized copanlisib-containing solid according to any one of claims 1 to 5, wherein copanlisib is present in an amount of 30.8 w/w% of said lyophilized copanlisib-containing solid, said citric acid is present in an amount of 2.98 w/w% of said lyophilized copanlisib-containing solid, said sodium hydroxide is present in an amount of 3.7 w/w% of said lyophilized copanlisib-containing solid, and said mannitol is present in an amount of 61.6 wt.%. /wt.% of the specified lyophilized solid containing copanlisib. 7. A lyophilized copanlisib-containing solid according to any one of claims 1 to 6, wherein copanlisib is present in an amount of 30, 45 or 60 mg, said citric acid is present in an amount of 5.8 mg, said sodium hydroxide is present in an amount of 6.3 mg and the said mannitol is present in an amount of 120 mg. 8. A lyophilized copanlisib-containing solid according to any one of claims 1 to 6, wherein copanlisib is present in an amount of 80 mg, said citric acid is present in an amount of 7.7 mg, said sodium hydroxide is present in an amount of 8.4 mg and said mannitol present in an amount of 160 mg. 9. A therapeutic, lyophilized, copanlisib-containing solid containing copanlisib in a therapeutically effective amount of 68.4 mg, one or more pH adjusters, a buffering agent containing a weak acid such as citric acid and a strong base such as sodium hydroxide , and a bulking agent, which is mannitol. 10. The lyophilized copanlisib-containing solid according to any one of claims 1 to 9, which is a powder or briquette. 11. A device which is a container containing a lyophilized copanlisib-containing solid according to any one of claims 1 to 10. 12. The device according to claim 11, which is a sealed container, preferably an injection vial, more preferably a 6 ml injection vial. -