Preparations containing melphalan flufenamide and cyclodextrin

Abstract

The present invention provides melphalan flufenamide or a salt thereof and γ-cyclodextrin or a pharma- ceutically acceptable derivative of γ-cyclodextrin. The present invention further provides a composition comprising the pharmaceutical preparation of the present invention, as well as a kit and a use of the pharmaceutical preparation of the present invention.

Description

[Technical field]

[0001] The present invention relates to novel pharmaceutical preparations comprising melphalan flufenamide or a salt thereof, and uses thereof. The present invention also provides kits and methods for preparing the novel pharmaceutical preparations of the present invention. [Background technology]

[0002] Melphalan flufenamide (also known as melflufen, L-melphalanyl-4-fluoro-L-phenylalanine ethyl ester and ethyl (2S)-2-[[(2S)-2-amino-3-[4-[bis(2-chloroethyl)amino]phenyl]propanoyl]amino]-3-(4-fluorophenyl)propanoate) is an antitumor agent useful in the treatment of cancer, particularly multiple myeloma. Melflufen is described in WO 01 / 96367 and WO 2014 / 065751.

[0003] In February 2021, the U.S. Food and Drug Administration (FDA) approved the use of melphalan flufenamide hydrochloride in the treatment of adult patients with relapsed or refractory multiple myeloma. In August 2022, the European Medicines Agency granted marketing authorization for the combination of melphalan flufenamide hydrochloride and dexamethasone for the treatment of adult patients with multiple myeloma who have received at least three prior lines of therapy and whose disease is refractory to at least one proteasome inhibitor, one immunomodulator, and one anti-CD38 monoclonal antibody, and who have shown disease progression on or after their last therapy, in countries of the European Union and the European Economic Area (EEA). Melphalan flufenamide hydrochloride is marketed in the EEA under the trade name PEPAXTI®. It is marketed in the United States under the trade name PEPAXTO®. The structure of melphalan flufenamide hydrochloride is shown below.

[0004] [ka]

[0005] PEPAXTI® is a white to off-white lyophilized powder. A single vial of PEPAXTI® contains 20 mg of melphalan flufenamide (21.48 mg of melphalan flufenamide hydrochloride) and 1,000 mg of sucrose. PEPAXTO® has the same formulation.

[0006] Further lyophilized preparations of melphalan flufenamide are described in WO 2012 / 146625 and WO 2014 / 065751. Liquid preparations of melphalan flufenamide comprising propylene glycol and / or polyethylene glycol as solvents are described in WO 2020 / 212594.

[0007] Prior to administration of PEPAXTI® (or PEPAXTO®), the lyophilized powder must be reconstituted with a 5% glucose solution (w / v in water) and then further diluted to the required concentration with a 0.9% sodium chloride solution (w / v in water) suitable for administration to patients by intravenous infusion. However, like many alkylating drugs, melphalan flufenamide is unstable in aqueous solutions. Therefore, to ensure that patients receive a safe and accurate dose of melphalan flufenamide, it is necessary to rapidly prepare the infusion solution and begin infusion to the patient immediately after the reconstitution step.

[0008] Improvements to the stability and / or handling characteristics of melphalan flufenamide preparations, particularly in aqueous solutions, would be advantageous in the clinic, for example allowing greater flexibility in storage conditions of melphalan flufenamide and the procedures used for preparation and administration of injection and infusion solutions. Summary of the Invention

[0009] The present invention provides melphalan flufenamide or a salt thereof, and γ-cyclodextrin (γCD) or a pharma- ceutically acceptable derivative of γ-cyclodextrin.

[0010] The inventors have found that melphalan flufenamide is surprisingly stable and soluble in aqueous solutions containing γCD. As described herein, in a stability test using melphalan flufenamide hydrochloride (Example compound 1, Ex1), the inventors found that after storing a solution containing melphalan flufenamide and γCD at 25°C-30°C for 22 hours, the melphalan flufenamide content of the solution was reduced by only 31% relative to the melphalan flufenamide content before storage of the solution at 25°C-30°C (i.e., t0). In the absence of any excipients, melphalan flufenamide hydrochloride is known to degrade at a rate of about 8%-10% per hour in aqueous solutions when stored at room temperature. A deuterated analog of melphalan flufenamide (Example compound 2, Ex2) was also found to exhibit improved stability in the presence of γCD.

[0011] In further stability studies, the inventors found that the melflufenflufenamid content in saline injection solutions containing melflufenflufenamid and γCD decreased by about 1% over 24 hours at 5° C. and about 2% over 48 hours at 5° C., whereas the melflufenflufenamid content in saline injection solutions without γCD decreased by about 5% over 24 hours at 5° C. and about 8% over 48 hours at 5° C. (see FIG. 3). That is, melflufenflufenamid showed a decomposition rate of about 1% per 24 hours at 5° C. in saline injection solutions containing γCD and a decomposition rate of about 4-5% per 24 hours at 5° C. in saline injection solutions without γCD.

[0012] The inventors also found that the positive effect of γCD can be obtained using pharma- ceutical acceptable derivatives of γ-cyclodextrin. In a further stability test, the inventors found that the melflufenfluramide content in saline injection solutions containing melflufenfluramide and hydroxypropyl-γCD decreased by about 4% on average over 5 hours at room temperature, while the melflufenfluramide content in saline injection solutions without HPγCD decreased by about 40% on average over the same period and under the same conditions. Similar results were found in glucose injection solutions. Increased stability was also seen for liquid formulations of melflufenfluramide and HPγCD in propylene glycol, compared to the same liquid formulations without any HPγCD.

[0013] In addition to improved stability, the pharmaceutical preparation of melphalan flufenamide with HPγCD allows for a higher reconstitution concentration of up to 3.0 mg / mL compared to the PEPTAXI® approved formulation, which has a limited reconstitution concentration of 0.5 mg / mL. Because a higher concentration of melphalan flufenamide can be achieved with the pharmaceutical preparation of the present invention, less liquid is required for initial reconstitution. This has the advantage that solid formulation pharmaceutical preparations, such as lyophilized pharmaceutical preparations, of melphalan flufenamide (or its salts) and γCD or pharma- ceutical acceptable γCD derivatives (such as HPγCD) can be provided in smaller vials.

[0014] Finally, the inventors have found that improved stability of melphalan flufenamine is achieved regardless of how γCD or a pharma- ceutically acceptable derivative of γCD, such as HPγCD, is formulated with melphalan flufenamine, which may be lyophilized with γCD or a pharma- ceutically acceptable derivative of γCD, such as HPγCD, which may be provided separately and mixed with melphalan flufenamide upon initial reconstitution (provided in a separate vial to melphalan flufenamide and mixed with it upon initial reconstitution to provide a 0.5-3.0 mg / mL solution before further dilution to an injection concentration of about 0.05 or 0.10 mg / mL), or which may be added to or provided in an injection solution (e.g., a physiologically acceptable solution having a concentration of melphalan flufenamide of 0.10-0.16 mg / mL that can be administered directly to a patient).

[0015] Typically, for an infusion solution of melflufenfluramide to be considered suitable for administration to a patient, the melflufenfluramide content in the solution when it is administered to a patient must be greater than 90% of the intended melflufenfluramide content when the solution was prepared. Thus, the reduction in the degradation rate of melflufenfluramide in the presence of γCD or a pharma- ceutically acceptable derivative of γCD allows for longer storage of the infusion solution prior to administration to a patient and also ensures that the quality of the infusion solution is not substantially reduced during the infusion process.

[0016] Furthermore, stabilized aqueous solutions of melphalan flufenamide are particularly useful in the clinic because they allow for greater flexibility and robustness in drug preparation, storage and administration protocols. For example, by increasing the stability of melphalan flufenamide in an aqueous infusion solution, the burden on clinical staff is dramatically reduced because the time spent preparing the drug for administration can be more flexible and when the drug can be administered. Furthermore, the ability to store an aqueous solution of melphalan flufenamide for longer periods of time allows for the preparation of larger batches of the aqueous infusion solution, which may then be transported to multiple clinical sites for administration to patients, and / or allows for staggering of patient appointments over a longer period of time without the need to make new batches of the aqueous infusion solution for patient groups arriving at different times. Such features are particularly important for highly potent cytotoxic agents such as melphalan flufenamide, because they reduce the exposure of clinical staff to the cytotoxic agent (i.e., because batches need to be prepared less frequently).

[0017] Melflufenflufenamide has also been found to exhibit a favorable dissolution rate in saline solutions containing γCD or pharma- ceutically acceptable derivatives of γCD, which is particularly beneficial in the clinic, since it allows the solution to be prepared quickly and eliminates the need for additional solvents or diluents to improve the dissolution rate of melflufenflufenamide in infusion solutions.

[0018] The invention also provides a composition comprising the pharmaceutical preparation of the invention and a physiologically acceptable aqueous solvent.

[0019] The present invention also provides a kit comprising melphalan flufenamide and a pharma- ceutically acceptable derivative of γCD or γ-cyclodextrin, and optionally a physiologically acceptable aqueous solvent.

[0020] The present invention also provides a pharmaceutical preparation or composition of the invention for use as a medicament.

[0021] The present invention also provides a pharmaceutical preparation or composition of the invention for use in the treatment or prevention of a disease or condition that can be treated with conventional chemotherapeutic agents, such as alkylating agents (e.g., melphalan, cyclophosphamide and bendamustine).

[0022] The present invention further provides a pharmaceutical preparation or composition of the invention for use in the treatment or prevention of cancer, such as a cancer selected from the list consisting of multiple myeloma, breast cancer, lung cancer, ovarian cancer, glioblastoma, leukemia and lymphoma.

[0023] The present invention further provides a pharmaceutical preparation or composition of the invention for use in the treatment or prevention of amyloidosis.

[0024] The present invention also provides a method for treating a patient, comprising administering to a subject in need thereof a pharma- ceutically effective amount of the pharmaceutical preparation or composition of the present invention. [Brief description of the drawings]

[0025] [Figure 1] The change in content of Example Compound 2 (Ex2) in saline solution containing Ex2 and γCD (top line of graph) and glucose solution containing Ex2 and γCD (second line from the top of graph) over 18 hours at 25° C. to 30° C. The change in content of Ex2 in saline and glucose solutions without γCD is also shown (third and fourth lines from the top of graph, respectively). For the avoidance of doubt, γCD is referred to as "gamma CD" in FIG. 1 and melphalan flufenamide is referred to as "melflufen". [Diagram 2]2 shows the change in melphalan flufenamide content in glucose solution containing melphalan flufenamide and γCD over 18 hours at room temperature. The melphalan flufenamide solution used in the experiment was prepared using a lyophilized melphalan flufenamide preparation containing melphalan flufenamide and sucrose. For the avoidance of doubt, γCD is referred to as "gCD" and "gamma CD" in FIG. 2, and melphalan flufenamide is referred to as "melflufen." [Diagram 3] 3 shows the change in melphalan flufenamide content in a melphalan flufenamide infusion solution over 43 hours at 5° C. The infusion solution was prepared in accordance with the guidance set forth in the US Prescribing Information for the melphalan flufenamide preparation sold under the name PEPTAXTO® dated February 2021. The top line of the graph shows the change in melphalan flufenamide content in the infusion solution containing γCD (at a γCD to melphalan flufenamide weight ratio (w / w) of 16:1) and the bottom line of the graph shows the change in melphalan flufenamide content in the infusion solution without γCD. For the avoidance of doubt, γCD is referred to as “gCD” in FIG. 3 and melphalan flufenamide is referred to as “melflufen.” DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0026] The present invention provides a pharmaceutical preparation comprising melflufen or a salt thereof and γ-cyclodextrin (γCD) or a pharma- ceutical acceptable derivative of γ-cyclodextrin. As described in the Examples section, the pharmaceutical preparation of the present invention exhibits excellent stability in aqueous solution.

[0027] Melphalan flufenamide For the avoidance of doubt, where the term "melphalan flufenamide" is used in this document, it is intended to include salts thereof, even if not expressly stated.

[0028] Also, for the avoidance of doubt, when the term "melphalan flufenamide" is used in this document, it includes isotopic derivatives of melphalan flufenamide unless otherwise specified. A specific example of an isotopic derivative of melphalan flufenamide suitable for use in the present invention is a deuterated melphalan flufenamide derivative. Examples of deuterated melflufenamide derivatives and methods for making deuterated melphalan flufenamide derivatives are described in WO2020 / 079165, the contents of which are incorporated herein by reference.

[0029] In certain embodiments, the deuterated melphalan flufenamide derivative suitable for use in the present invention is 2 H5) Ethyl (2S)-2-[(2S)-2-amino-3-{4-[bis(2-chloroethyl)amino]phenyl}propanamido]-3-(4-fluorophenyl)propanoate hydrochloride (hereinafter referred to as Ex2). Ex2 has the following structure:

[0030] [ka] has.

[0031] The inventors have found that the pharmaceutical preparation of the present invention is particularly suitable for isotopic derivatives of melphalan flufenamide, such as Ex2 (see Example 3 and Figure 1 of the present application). Thus, in a preferred embodiment, the pharmaceutical preparation of the present invention comprises Ex2 or a salt thereof, and a pharma- ceutically acceptable derivative of γCD or γ-cyclodextrin, such as HPγCD. In another preferred embodiment, the pharmaceutical preparation of the present invention comprises Ex2 or a salt thereof, and γCD. The inventors have also found that the pharmaceutical preparation of the present invention is particularly suitable for melphalan flufenamide with natural isotopic abundance. Thus, in another preferred embodiment, the pharmaceutical preparation of the present invention comprises melphalan flufenamide with natural isotopic abundance, or a salt thereof, and a pharma- ceutically acceptable derivative of γCD or γ-cyclodextrin, such as HPγCD.

[0032] Also, for the avoidance of doubt, when referred to in this document, the mass of melphalan flufenamide is the mass of the melphalan flufenamide molecule excluding the mass of any counterion (i.e., melphalan flufenamide as its free base), unless expressly stated otherwise. The molecular weight of melflufen (also referred to herein as "free base") (with natural isotopic abundance) without counterion is 498.42 g / mol. Melphalan flufenamide and its salts, in particular its hydrochloride salt, are known, for example, from WO 01 / 96367, WO 2014 / 065751 and WO 2020 / 212594, and the same salts are suitable for use in the present invention. The salts of melphalan flufenamide suitable for use in the present invention are those in which the counterion is pharma- ceutically acceptable. Suitable salts include those formed with organic or inorganic acids. In particular, suitable salts formed with acids according to the invention include those formed with mineral acids such as amino acids, strong organic carboxylic acids, such as hydroxycarboxylic acids, such as alkanecarboxylic acids of 1 to 4 carbon atoms, either unsubstituted or substituted, for example, by halogens, or with organic sulfonic acids, such as (C1-C4) alkyl or aryl sulfonic acids, either unsubstituted or substituted, for example, by halogens. Pharmaceutically acceptable acid addition salts include those formed from hydrochloric, hydrobromic, sulfuric, nitric, citric, tartaric, acetic, phosphoric, lactic, pyruvic, acetic, trifluoroacetic, succinic, perchloric, fumaric, maleic, glycolic, lactic, salicylic, oxalic, oxaloacetic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic, benzenesulfonic, isethionic, ascorbic, malic, phthalic, aspartic, and glutamic acids, lysine, and arginine.

[0033] Preferred salts of melphalan flufenamide include acid addition salts such as those formed from hydrochloric acid, hydrobromic acid, acetic acid, p-toluenesulfonic acid, tartaric acid, sulfuric acid, succinic acid, phosphoric acid, oxalic acid, nitric acid, methanesulfonic acid, malic acid, maleic acid, and citric acid. Typically, the salt of melphalan flufenamide for use according to the invention is the hydrochloride salt (i.e., an addition salt formed from hydrochloric acid).

[0034] Other preferred salts of melphalan flufenamide include hydrochloric acid, hydrobromic acid, nitric acid, methanesulfonic acid, sulfuric acid, phosphoric acid, trifluoroacetic acid, para-toluenesulfonic acid, 2-mesitylsulfonic acid, citric acid, acetic acid, tartaric acid, fumaric acid, lactic acid, succinic acid, malic acid, malonic acid, maleic acid, 1,2-ethanedisulfonic acid, adipic acid, aspartic acid, benzenesulfonic acid, benzoic acid, ethanesulfonic acid or nicotinic acid.

[0035] Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with the solvents with which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates." For example, a complex with water is known as a "hydrate." The complex can incorporate the solvent in stoichiometric or non-stoichiometric amounts. Solvates are described in Water-Insoluble Drug Formulation, 2 nd ed R. Lui CRC Press, page 553 and Byrn et al Pharm Res 12(7), 1995, 945-954. Before being made into a solution, melphalan flufenamide, or a salt thereof, for use in the present invention may be in the form of a solvate. Solvates of melphalan flufenamide suitable for use in the present invention are those in which the associated solvent is pharma- ceutically acceptable. For example, hydrates are pharma- ceutically acceptable solvates.

[0036] Pharmaceutical Preparations The present invention provides melphalan flufenamide or a salt thereof and a pharma- ceutically acceptable derivative of γCD or γ-cyclodextrin.

[0037] γCD is a cyclic oligosaccharide composed of eight α-1,4-linked glucopyranose units. It is a known excipient for pharmaceutical and nutritional products and has been used as a delivery vehicle for hydrophobic or poorly water-soluble molecules.

[0038] Various pharma- ceutically acceptable derivatives of γ-cyclodextrin are known. Preferably, such derivatives are soluble in water, for example, to a concentration of at least 5% by weight (i.e., at least 50 g / L), for example, at least 10% by weight, at room temperature. Examples of pharma- ceutically acceptable derivatives of γ-cyclodextrin include, but are not limited to, hydroxypropyl-γ-cyclodextrin, succinyl-γ-cyclodextrin, sulfobutylether-γ-cyclodextrin, sodium γ-cyclodextrin phosphate, sodium γ-cyclodextrin sulfate, 2-carboxyethyl-γ-cyclodextrin, acetylated-γ-cyclodextrin, and sugammadex. For example, hydroxypropyl-γCD (HPγCD) is highly suitable for use in pharmaceutical preparations, particularly in the pharmaceutical preparations, formulations, and compositions of the present invention. HPγCD is a particularly preferred pharma- ceutically acceptable derivative of γCD according to the present invention. In this specification, unless the context requires otherwise, comments regarding γCD apply equally to derivatives of γCD (and in particular to pharma- ceutically acceptable derivatives of γCD, such as HPγCD, succinyl-γ-cyclodextrin, sulfobutylether-γ-cyclodextrin, γ-cyclodextrin sodium phosphate, γ-cyclodextrin sodium sulfate, 2-carboxyethyl-γ-cyclodextrin, acetylated-γ-cyclodextrin and sugammadex, in particular HPγCD).

[0039] For the avoidance of doubt, in embodiments in which the pharmaceutical preparation of the present invention comprises a pharma- ceutically acceptable derivative of γ-cyclodextrin, the pharma- ceutically acceptable derivative of γ-cyclodextrin may have any degree of substitution (DS, also known as molar substitution). As used herein, DS is the average number of derivative group substituents (e.g., the average number of hydroxypropyl, sulfobutyl ether, 2-carboxyethyl, acetyl and / or 3-sulfanylpropanoic acid substituents) that have reacted with one glucopyranose repeat unit of γCD. Each glucopyranose repeat unit has three hydroxyl groups that can be replaced by derivative group substituents. The DS number represents the average DS of the mixture of isomers. For some derivative group substituents (such as the hydroxypropyl group), the electrophile used to react with γCD to form a derivative of γCD (e.g., propylene oxide) can react with the hydroxyl groups of the derivative group substituent itself and with the hydroxyl groups of the glucopyranose repeat unit to form a polymeric side chain (e.g., polypropylene glycol). Thus, the DS value for a derivative of γ-cyclodextrin can be greater than 3 when two or more substituents react to form oligomeric or polymeric side chains.

[0040] As an illustrative example, a pharma- ceutically acceptable derivative of γ-cyclodextrin of the invention having a DS of about 0.6 (e.g., HPγCD) has, on average, 0.6 derivative group substituents (e.g., hydroxypropyl groups) reacted with one glucopyranose repeat unit of γCD. Because γCD has eight glucopyranose repeat units, a pharma- ceutically acceptable derivative of γ-cyclodextrin defined as having a DS of 0.6 has, on average, about 4.7 (e.g., 4.7 or 4.8) derivative group substituents (e.g., hydroxypropyl groups) per γCD molecule.

[0041] In certain embodiments, the pharmaceutical preparation of the invention comprises a pharma- ceutically acceptable derivative of γ-cyclodextrin having a DS of at least 0.04, e.g., from about 0.04 to about 5. For example, from about 0.04 to about 4, from about 0.04 to about 3, or from about 0.04 to about 3. In certain embodiments, the pharmaceutical preparation of the invention comprises a pharma- ceutically acceptable derivative of γ-cyclodextrin having a DS of at least 0.08, 0.1, at least 0.2, at least 0.4, or at least 0.5. In certain embodiments, the pharmaceutical preparation of the invention comprises a pharma- ceutically acceptable derivative of γ-cyclodextrin having a DS of less than 5, less than 4, less than 3, less than 2, less than 1.5, less than 1, or less than 0.8. In a particular embodiment, the pharma- ceutically acceptable derivative of γ-cyclodextrin of the present invention has a DS of about 0.04 to about 2, for example, about 0.1 to about 1.5, about 0.1 to about 1, about 0.2 to about 1, about 0.2 to about 0.8, or about 0.4 to about 0.8. In a preferred embodiment, the pharma- ceutically acceptable derivative of γ-cyclodextrin is HPγCD having a DS of about 0.04 to about 1, more preferably about 0.1 to about 1, more preferably about 0.2 to about 0.8, and most preferably 0.4 to 0.8, for example, 0.4, 0.5, 0.6, 0.7, or 0.8. In a preferred embodiment, the pharma- ceutically acceptable derivative of γ-cyclodextrin is HPγCD having a DS of about 0.6.

[0042] The pharmaceutical preparation of the present invention comprises γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) in a weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 1:1 to about 100:1. Typically, the pharmaceutical preparation comprises γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) in a weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 4:1 to about 80:1.

[0043] Melphalan flufenamide exhibits particularly good stability in aqueous solution when γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) is included in the pharmaceutical preparation at a weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 4:1 to about 25:1. For example, the inventors have shown that an aqueous solution of melphalan flufenamide containing γCD at a weight ratio (w / w) of γCD to melphalan flufenamide of about 4:1 retained 69% of melphalan flufenamide after 22 hours at 25° C. to 30° C. In further stability tests, the inventors have shown that melphalan flufenamide exhibits particularly good stability in aqueous solutions containing γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) at weight ratios (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 10:1 to about 50:1, about 10:1 to about 30:1, about 10:1 to about 28:1, about 10:1 to about 25:1, and about 10:1 to about 23:1.

[0044] In further stability tests, the inventors have shown that melphalan flufenamide exhibits particularly good stability in aqueous solutions containing γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) at weight ratios (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 10:1 to about 50:1, about 10:1 to about 30:1, about 10:1 to about 28:1, about 10:1 to about 25:1, and about 10:1 to about 23:1. In certain stability studies, the inventors have shown that melphalan flufenamide exhibits particularly good stability in aqueous solutions containing γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) at weight ratios (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 15:1 to about 50:1, about 15:1 to about 30:1, about 15:1 to about 28:1, and about 15:1 to about 25:1.

[0045] Thus, preferably, the pharmaceutical preparation of the invention comprises γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) in a weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 4:1 to about 25:1. In an exemplary embodiment, the pharmaceutical preparation of the invention may comprise γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) in a weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of at least about 5:1. In certain embodiments, the pharmaceutical preparation of the invention comprises γCD (or a pharma- ceutical acceptable derivative of γ-cyclodextrin, e.g., HPγCD) in a weight ratio (w / w) of γCD (or a pharma- ceutical acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 5:1 to about 25:1. More preferably, the weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide is about 5:1 to about 25:1, e.g., about 10:1 to about 25:1 (e.g., a pharmaceutical preparation of the present invention may contain γCD in a weight ratio (w / w) of γCD to melphalan flufenamide of about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1 or about 25:1). In one preferred embodiment, the weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide is about 10:1 to about 23:1 (e.g., the weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide is about 10:1, about 15:1, about 18:1, or about 23:1).

[0046] In an exemplary embodiment in which the pharmaceutical preparation of the present invention comprises γCD in a weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 5:1 to about 25:1, the pharmaceutical preparation may comprise, for example, 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion) and about 5 mg to about 750 mg of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD). For example, a pharmaceutical preparation comprising 20 mg of melphalan flufenamide (excluding the mass of any counterion) may comprise about 100 mg to about 500 mg of γCD (e.g., about 200 mg to about 500 mg of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD)).

[0047] In another exemplary embodiment in which the pharmaceutical preparation of the present invention comprises γCD in a weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 15:1 to about 25:1, the pharmaceutical preparation may comprise, for example, 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion) and about 15 mg to about 750 mg of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD). For example, a pharmaceutical preparation comprising 20 mg of melphalan flufenamide (excluding the mass of any counterion) may comprise about 300 mg to about 500 mg of γCD (e.g., about 320 mg to about 500 mg of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD)).

[0048] In another exemplary embodiment in which the pharmaceutical preparation of the present invention comprises γCD in a weight ratio (w / w) of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD) to melphalan flufenamide of about 20:1 to about 25:1, the pharmaceutical preparation may comprise, for example, 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion) and about 20 mg to about 750 mg of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD). For example, a pharmaceutical preparation comprising 20 mg of melphalan flufenamide (excluding the mass of any counterion) may comprise about 400 mg to about 500 mg of γCD (e.g., about 400 mg, about 420 mg, about 440 mg, about 460 mg, about 480 mg, or about 500 mg of γCD (or a pharma- ceutically acceptable derivative of γ-cyclodextrin, e.g., HPγCD)).

[0049] Melphalan flufenamide exhibits particularly good stability in aqueous solution when HPγCD is included in the pharmaceutical preparation at a weight ratio (w / w) of HPγCD to melphalan flufenamide of about 10:1 to about 50:1, e.g., about 20:1 to about 40:1 or about 15:1 to about 25:1. For example, the inventors have shown that an aqueous solution of melphalan flufenamide containing HPγCD at a weight ratio (w / w) of HPγCD to melphalan flufenamide of about 36:1 retained 87% of melphalan flufenamide after 18 hours at room temperature. As a further example, the inventors have shown that aqueous solutions of 0.2 mg / mL melphalan flufenamide containing HPγCD in a weight ratio (w / w) of HPγCD to melphalan flufenamide of 1:20, 1:24, 1:28, 1:32, 1:36 or 1:40 retained greater than 78% of the melphalan flufenamide content after 18 hours at room temperature and greater than 93% of the melphalan flufenamide content after 18 hours after refrigeration. As another example, the inventors have shown that aqueous solutions of 0.1 and 0.5 mg / mL melphalan flufenamide containing HPγCD in a weight ratio (w / w) of HPγCD to melphalan flufenamide of 1:16, 1:20, 1:24 or 1:28 retained greater than 80% and 90% of the melphalan flufenamide content, respectively, after 20 hours at room temperature.

[0050] In exemplary embodiments in which a pharmaceutical preparation of the invention comprises HPγCD in a weight ratio (w / w) of HPγCD to melphalan flufenamide of about 15:1 to about 40:1, the pharmaceutical preparation may comprise, for example, 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion) and about 15 mg to about 1200 mg of HPγCD. For example, a pharmaceutical preparation comprising 20 mg of melphalan flufenamide (excluding the mass of any counterion) may comprise about 300 mg to about 800 mg of HPγCD (e.g., about 500 mg to about 700 mg of HPγCD).

[0051] In exemplary embodiments in which a pharmaceutical preparation of the invention comprises HPγCD in a weight ratio (w / w) of HPγCD to melphalan flufenamide of about 20:1 to about 40:1, the pharmaceutical preparation may comprise, for example, 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion) and about 20 mg to about 1200 mg of HPγCD. For example, a pharmaceutical preparation comprising 20 mg of melphalan flufenamide (excluding the mass of any counterion) may comprise about 400 mg to about 800 mg of HPγCD (e.g., about 500 mg to about 700 mg of HPγCD). In further exemplary embodiments in which the pharmaceutical preparation of the invention comprises HPγCD in a weight ratio (w / w) of HPγCD to melphalan flufenamide of about 32:1 to about 40:1, the pharmaceutical preparation may comprise, for example, 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion) and about 32 mg to about 1200 mg of HPγCD. For example, a pharmaceutical preparation comprising 20 mg of melphalan flufenamide (excluding the mass of any counterion) may comprise about 640 mg to about 800 mg of HPγCD (e.g., about 680 mg to about 760 mg of HPγCD).

[0052] In further preferred exemplary embodiments in which the pharmaceutical preparation of the invention comprises HPγCD in a weight ratio (w / w) of HPγCD to melphalan flufenamide of about 15:1 to about 25:1, the pharmaceutical preparation may, for example, comprise 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion) and about 15 mg to about 750 mg of HPγCD. For example, a pharmaceutical preparation comprising 20 mg of melphalan flufenamide (excluding the mass of any counterion) may comprise about 300 mg to about 500 mg of HPγCD (e.g., about 300 mg, about 320 mg, about 360 mg, about 400 mg, about 440 mg, about 480 mg, or about 500 mg of HPγCD).

[0053] In further preferred exemplary embodiments in which the pharmaceutical preparation of the invention comprises HPγCD in a weight ratio (w / w) of HPγCD to melphalan flufenamide of about 20:1 to about 25:1, the pharmaceutical preparation may comprise, for example, 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion) and about 20 mg to about 750 mg of HPγCD. For example, a pharmaceutical preparation comprising 20 mg of melphalan flufenamide (excluding the mass of any counterion) may comprise about 400 mg to about 500 mg of HPγCD (e.g., about 400 mg, about 420 mg, about 440 mg, about 460 mg, about 480 mg, or about 500 mg of HPγCD).

[0054] Additional excipients may be included in the pharmaceutical preparation of the present invention. Additional excipients may be included in the pharmaceutical preparation to improve the dissolution rate of melphalan flufenamide in aqueous solution and / or to improve the stability of melphalan flufenamide, for example, when in its lyophilized form. Notably, the inventors have found that γCD and pharma- ceutically acceptable derivatives of γCD (such as HPγCD) have a stabilizing effect on melphalan flufenamide even when additional excipients are included in the pharmaceutical preparation. Thus, in certain embodiments, the pharmaceutical preparation of the present invention comprises one or more additional pharma- ceutically acceptable excipients in addition to γCD or pharma- ceutically acceptable derivatives of γ-cyclodextrin (such as HPγCD).

[0055] The one or more further pharma- ceutically acceptable excipients may be selected from the group consisting of polysorbate 80, propylene glycol, sucrose, trehalose dehydrate, MgCl2 and CaCl2. Preferably, the further pharma- ceutically acceptable excipient is sucrose.

[0056] Typically, when a pharmaceutical preparation of the invention includes, in addition to γCD, one or more additional excipients, the one or more additional excipients are each independently present in the preparation in a weight ratio (w / w) of excipient to melphalan flufenamide of about 1:1 to about 100:1, e.g., about 1:1 to about 50:1.

[0057] In certain embodiments, the pharmaceutical preparation of the present invention is a lyophilized powder. The inventors have found that γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) can be included in the lyophilized melphalan flufenamide formulation. Reconstitution of the lyophilized powder provides an aqueous solution in which melphalan flufenamide is particularly stable.

[0058] The pharmaceutical preparation of the present invention, when in the form of a lyophilized powder, may consist essentially of melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD). Alternatively, the pharmaceutical preparation of the present invention, when in the form of a lyophilized powder, may comprise, in addition to γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD), one or more further excipients, and in particular, in addition to γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD), one further excipient. The further excipient may be selected from the group consisting of sucrose, polysorbate 80, propylene glycol, anhydrous trehalose, MgCl2 and CaCl2. Preferably, the further excipient is sucrose.

[0059] In an exemplary embodiment in which the pharmaceutical preparation is a lyophilized powder, the pharmaceutical preparation comprises (or consists essentially of) melphalan flufenamide, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD), and sucrose. Sucrose has been found to improve the dissolution rate of melphalan flufenamide in aqueous solutions. When sucrose is present in the pharmaceutical preparation of the present invention, it is typically present in a weight ratio (w / w) of sucrose to melphalan flufenamide of about 1:1 to about 500:1. For example, sucrose may be present in a weight ratio (w / w) of sucrose to melphalan flufenamide of about 2:1 to about 100:1, about 1:2 to about 1:75, or about 1:4 to about 1:75. Preferably, sucrose is present in a weight ratio (w / w) of sucrose to melphalan flufenamide of about 20:1 to about 75:1. More preferably, the sucrose is present in a weight ratio (w / w) of sucrose to melphalan flufenamide of from about 25:1 to about 75:1. Even more preferably, the sucrose is present in a weight ratio (w / w) of sucrose to melphalan flufenamide of from about 30:1 to about 70:1, from about 40:1 to about 60:1, or from about 45:1 to about 55:1 (e.g., 50:1).

[0060] In certain embodiments where the pharmaceutical preparation is a lyophilized powder, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) to melphalan flufenamide is about 4:1 to about 80:1 (e.g., about 5:1 to about 25:1), and the weight ratio (w / w) of sucrose to melphalan flufenamide is about 25:1 to 75:1 (e.g., 50:1). For example, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) to melphalan flufenamide is about 10:1 to about 25:1, and the weight ratio (w / w) of sucrose to melphalan flufenamide is about 25:1 to 75:1 (e.g., 50:1). An exemplary embodiment of such a preparation comprises 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion), about 5 mg to about 750 mg of γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) and about 0.25 g to 2.25 g of sucrose. In a further exemplary embodiment, such a preparation may comprise 20 mg of melphalan flufenamide (excluding the mass of any counterion), 1 g of sucrose, and about 200 mg to about 500 mg of γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) (e.g., 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, or 500 mg of γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD)).

[0061] Preferably, in embodiments where the pharmaceutical preparation is a lyophilized powder, the pharmaceutical preparation is free or substantially free of any organic solvent. By "free" of organic solvent, it is meant that the lyophilized powder pharmaceutical preparation does not contain any measurable amount of organic solvent. By "substantially free" of any organic solvent, it is meant that the lyophilized powder pharmaceutical preparation contains only trace amounts of organic solvent, e.g., less than about 0.1% total w / v organic solvent.

[0062] In certain embodiments, the pharmaceutical preparation of the present invention is a liquid pharmaceutical formulation, i.e., the formulation is in liquid form when stored at room temperature.

[0063] In an embodiment where the pharmaceutical preparation is a liquid pharmaceutical preparation, the pharmaceutical preparation comprises melphalan flufenamide, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD), and a pharma- ceutically acceptable organic solvent. Typically, the pharma- ceutically acceptable organic solvent is selected from the group consisting of propylene glycol, polyethylene glycol (PEG), and mixtures thereof. In one embodiment, the pharmaceutical preparation is a liquid pharmaceutical preparation, and the pharmaceutical preparation comprises (or consists of) melphalan flufenamide, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD), and propylene glycol.

[0064] In embodiments where the pharmaceutical preparation is a liquid pharmaceutical preparation, the pharmaceutical preparation may optionally contain one or more physiologically acceptable aqueous solvents. Suitable aqueous solvents include non-toxic parenterally acceptable diluents or solvents, such as water, Ringer's solution, glucose solution, and sodium chloride solution (e.g., isotonic sodium chloride solution). The pharmaceutical preparation may optionally further contain additional components, such as antioxidants, buffers, bacteriostats, and solutes that render the pharmaceutical preparation of the present invention isotonic with the blood of the intended recipient.

[0065] In an alternative embodiment where the pharmaceutical preparation is a liquid pharmaceutical formulation, the pharmaceutical preparation may optionally be substantially free of a physiologically acceptable aqueous solvent. In a further alternative embodiment where the pharmaceutical preparation is a liquid pharmaceutical formulation, the pharmaceutical preparation may optionally be substantially free of an aqueous solvent. By "substantially free" of a physiologically acceptable aqueous solvent or solvent, it is meant that the pharmaceutical preparation liquid formulation contains only trace amounts of a physiologically acceptable aqueous solvent or solvent. For example, less than about 1% w / v of a physiologically acceptable aqueous solvent or solvent. Preferably, less than 0.5% w / v of a physiologically acceptable aqueous solvent or solvent, and more preferably less than 0.1% w / v of a physiologically acceptable aqueous solvent or solvent.

[0066] In certain embodiments, where the pharmaceutical preparation is a liquid pharmaceutical formulation, the pharmaceutical preparation consists essentially of melphalan flufenamide or a salt thereof, γCD, and propylene glycol.

[0067] In other embodiments, where the pharmaceutical preparation is a liquid pharmaceutical formulation, the pharmaceutical preparation consists essentially of melphalan flufenamide or a salt thereof, γCD and PEG.

[0068] In certain embodiments, where the pharmaceutical preparation is a liquid pharmaceutical formulation, the pharmaceutical preparation consists essentially of melphalan flufenamide or a salt thereof, HPγCD and propylene glycol.

[0069] In other embodiments, where the pharmaceutical preparation is a liquid pharmaceutical formulation, the pharmaceutical preparation consists essentially of melphalan flufenamide or a salt thereof, HPγCD and PEG.

[0070] The pharmaceutical preparations of the invention comprising PEG typically comprise PEG having an average molecular weight of 150-650 Da, for example 150-250, 250-350, 350-450, 450-550 and 550-650 Da. For example, the PEG may be PEG200, PEG300, PEG400 or PEG600 as defined in the national pharmacopoeias. Preferably, the PEG has an average molecular weight of 250-350 Da. The pharmaceutical preparations of the invention may comprise a mixture of two or more different PEG compounds, for example a mixture comprising two or more of PEG200, PEG300, PEG400 and PEG600.

[0071] In other embodiments, the pharmaceutical preparation comprises melphalan flufenamide or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD), propylene glycol, and PEG. In certain embodiments, the pharmaceutical preparation consists essentially of melphalan flufenamide or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD), propylene glycol, and PEG.

[0072] For pharmaceutical preparations of the invention comprising PEG (e.g., PEG200, PEG300, PEG400, or PEG600) and propylene glycol, the weight ratio (w / w) of propylene glycol to PEG content can be, for example, about 1:1 to about 1:50 (e.g., 0.5:1, 1:1, 1:2, 1:5, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50) or about 1:1 to about 50:1 (e.g., 1:0.5, 2:1, 5:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, or 50:1).

[0073] In embodiments of the invention in which the pharmaceutical preparation comprises PEG and propylene glycol, the PEG content may consist of one PEG compound, as described above, or may consist of a mixture of two or more different PEG compounds, as described above.

[0074] Propylene glycol and PEG suitable for use according to the invention are readily available from commercial sources. It should be understood that the source and / or grade of propylene glycol, PEG and / or any other solvents present in the pharmaceutical preparation of the invention are not relevant to the beneficial effects of the invention to be realized.

[0075] In certain embodiments where the pharmaceutical preparation is a liquid formulation, the pharmaceutical preparation is substantially free of organic solvents that are not PEG or propylene glycol. By "substantially free" of organic solvents, it is meant that the pharmaceutical preparation liquid formulation contains only trace amounts of organic solvents other than propylene glycol and PEG. For example, less than about 1% w / v of organic solvents other than propylene glycol or PEG. Preferably, less than 0.5% w / v, and more preferably less than 0.1% w / v of organic solvents other than propylene glycol or PEG. In embodiments where the pharmaceutical preparation is a liquid formulation, the melphalan flufenamide may be present (e.g., is present) at a concentration of about 1 mg / mL to about 50 mg / mL (excluding the mass of any counterion). Typically, the melphalan flufenamide is at a concentration of about 10 mg / mL to about 30 mg / mL (excluding the mass of any counterion). For example, 15 mg / mL to 25 mg / mL or 17.5 mg / mL to 22.5 mg / mL (excluding the mass of any counterion). For example, such a liquid formulation may contain melphalan flufenamide at a concentration of 20 mg / mL (excluding the mass of any counterion) of melphalan flufenamide (and, for example, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin, such as HPγCD, at a concentration of about 200 mg / mL to about 500 mg / mL).

[0076] In embodiments where the pharmaceutical preparation is a liquid formulation, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) may be present (e.g., is present) at a concentration of about 1 mg / mL to about 5000 mg / mL (excluding the mass of any counterions). Typically, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) is at a concentration of about 40 mg / mL to about 1200 mg / mL (excluding the mass of any counterions). For example, about 60 to about 1000 mg / mL or about 70 mg / mL to about 900 mg / L (excluding the mass of any counterions). In one embodiment, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) is at a concentration of 100 mg / mL to 500 mg / mL or 150 mg / mL to 600 mg / L (excluding the mass of any counterions). In another embodiment, γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) is at a concentration of 200 mg / mL to 500 mg / mL (excluding the mass of any counterion). For example, such a liquid formulation contains melphalan flufenamide at a concentration of 15 mg / mL to 25 mg / mL or 17.5 mg / mL to 22.5 mg / mL (excluding the mass of any counterion) and γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD) at a concentration of about 200 mg / mL to about 500 mg / mL.

[0077] When the pharmaceutical preparation of the present invention is in the form of a liquid pharmaceutical formulation, it can be directly administered to a subject, for example, without prior dilution, for example, without prior dilution with a physiologically acceptable solvent or diluent.Therefore, in one preferred embodiment of the present invention, the pharmaceutical preparation is in a form suitable for direct administration to a subject without dilution.In another preferred embodiment, the pharmaceutical preparation is in a form suitable for administration to a patient after dilution, for example, after dilution with a physiologically acceptable solvent or diluent, for example, saline solution, glucose solution, or mixtures thereof.

[0078] In certain embodiments where the pharmaceutical preparation is a liquid pharmaceutical formulation, the pharmaceutical preparation preferably does not include any additional physiologically acceptable solvent. Alternatively, or in addition, in certain embodiments where the pharmaceutical preparation is a liquid pharmaceutical formulation, the pharmaceutical preparation preferably does not include any additional physiologically acceptable aqueous solvent.

[0079] In certain embodiments, where the pharmaceutical preparation is a liquid formulation, the pharmaceutical preparation is substantially free of water. By "substantially free of water" it is meant that the pharmaceutical preparation liquid formulation contains only trace amounts of water. For example, less than about 1% w / v water. Preferably, less than 0.5% w / v water, more preferably less than 0.1% w / v water.

[0080] The present invention also provides a composition comprising the pharmaceutical preparation of the present invention and a physiologically acceptable aqueous solvent or diluent. The composition of the present invention may be suitable for parenteral administration to a subject. As described in the Examples section, injection and infusion solutions may be prepared by diluting the lyophilized powder or liquid pharmaceutical preparation of the present invention with one or more physiologically acceptable solvents or diluents. Exemplary solvents or diluents include mannitol, glucose, 1,3-butanediol, water, Ringer's solution, isotonic sodium chloride solution, or other suitable dispersing or wetting agents and suspending agents, including synthetic mono- or diglycerides, and fatty acids, including oleic acid, or Cremaphor.

[0081] In certain embodiments, the pharmaceutical preparations of the present invention are administered to a subject, for example, after dilution with a physiologically acceptable aqueous solvent or diluent, such as a saline solution, a glucose solution, or a mixture thereof. Any such solvent or diluent may optionally be buffered.

[0082] Preferably, the physiologically acceptable solvent is an aqueous solution such as a glucose solution (e.g., a 5% glucose solution (w / v in water)), a saline solution (e.g., a 0.9% sodium chloride solution (w / v in water)), or a mixture thereof.

[0083] The pharmaceutical preparation of the present invention may be diluted with a physiologically acceptable solvent prior to administration to a subject to a melphalan flufenamide concentration (excluding the mass of any counterion), for example, of about 0.001 mg / mL to about 3 mg / mL (e.g., 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1.4 mg / mL, 0.05 mg / mL to 1.2 mg / mL, 0.05 mg / mL to 1 mg / mL, 0.05 mg / mL to 0.9 mg / mL, 0.05 mg / mL to 0.8 mg / mL, 0.1 mg / mL to 0.4 mg / mL, or 0.1 mg / mL to 0.3 mg / mL).

[0084] Thus, in certain embodiments, the pharmaceutical preparation of the invention comprises melphalan flufenamide at a concentration of about 0.001 mg / mL to about 3 mg / mL (e.g., 0.01 mg / mL to 2 mg / mL, 0.05 mg / mL to 1.4 mg / mL, 0.05 mg / mL to 1.2 mg / mL, 0.05 mg / mL to 1 mg / mL, 0.05 mg / mL to 0.9 mg / mL, 0.05 mg / mL to 0.8 mg / mL, 0.1 mg / mL to 0.4 mg / mL, 0.1 mg / mL to 0.3 mg / mL, or 0.1 mg / mL to 0.16 mg / mL) (excluding the mass of any counterion).

[0085] In certain preferred embodiments, the pharmaceutical preparations of the invention may be diluted with a physiologically acceptable solvent prior to administration to a concentration of, for example, about 0.05 mg / mL to 0.9 mg / mL, preferably 0.1 mg / mL to 0.3 mg / mL, and more preferably about 0.1 mg / mL to 0.16 mg / mL of melphalan flufenamide (excluding the mass of any counterion).

[0086] Melphalan flufenamide (e.g., melphalan flufenamide in its hydrochloride form (Ex1) or deuterated melflufenamide such as Ex2) may be administered to a subject at a concentration of about 0.1 mg / mL to about 3 mg / mL (excluding the mass of any counterion). Dosage regimens commonly used in the clinic for Ex1 require a melphalan flufenamide concentration in the infusion solution of 0.1 mg / mL to 0.16 mg / mL. Thus, in certain embodiments, the pharmaceutical preparation of the present invention comprises melphalan flufenamide at a concentration of about 0.1 mg / mL to about 0.16 mg / mL (excluding the mass of any counterion) (e.g., 0.10 mg / mL to 0.16 mg / mL).

[0087] Compositions suitable for parenteral administration according to the present invention are preferably suitable for administration by infusion or injection, particularly suitable for administration by intravenous infusion.

[0088] In certain embodiments, injection and infusion solutions of melphalan flufenamide can be prepared by diluting a lyophilized powder or liquid pharmaceutical formulation of melphalan flufenamide without γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) with one or more physiologically acceptable solvents or diluents, including γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD). For example, melphalan flufenamide can be a lyophilized powder that includes (or consists essentially of) melphalan flufenamide and sucrose. For example, the lyophilized powder can include (or consist essentially of) melphalan flufenamide and sucrose in a weight ratio of melphalan flufenamide to sucrose of 1:50 (e.g., one vial of lyophilized powder can include 20 mg of melphalan flufenamide (i.e., 21.48 mg of melphalan flufenamide hydrochloride) and 1,000 mg of sucrose). Alternatively, for example, melphalan flufenamide may be a liquid pharmaceutical formulation comprising (or consisting essentially of) melphalan flufenamide and propylene glycol and / or PEG. For example, the liquid pharmaceutical formulation comprises melphalan flufenamide at a concentration of 15 mg / mL to 25 mg / mL and propylene glycol and / or PEG as a solvent.

[0089] In embodiments in which the one or more physiologically acceptable solvents or diluents comprise γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), the γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) may be present at a concentration of about 1 mg / mL to 10 mg / mL. For example, the γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) may be present at a concentration of about 1 mg / mL to 3 mg / mL, e.g., about 1 mg / mL, 1.5 mg / mL, 2 mg / mL, 2.5 mg / mL, 3 mg / mL, 4 mg / mL, or 5 mg / mL.

[0090] Alternatively or additionally, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) may be added directly to the injection or infusion solution after reconstitution and / or dilution of melphalan flufenamide with a suitable physiologically acceptable solvent or diluent, or melphalan flufenamide may be added directly to the injection or infusion solution after reconstitution / dilution of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) with a suitable physiologically acceptable solvent or diluent. Thus, the present invention also provides an injection or infusion solution comprising melphalan flufenamide, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), one or more physiologically acceptable solvents or diluents, and optionally one or more further excipients.

[0091] In embodiments in which γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) is added directly to the injection or infusion solution after reconstitution and / or dilution of melphalan flufenamide with a suitable physiologically acceptable solvent or diluent, or in which melphalan flufenamide is added directly to the injection or infusion solution after reconstitution and / or dilution of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) with a suitable physiologically acceptable solvent or diluent, the melphalan flufenamide may be in the form of a pharmaceutical preparation comprising melphalan flufenamide, for example, a lyophilized powder or liquid pharmaceutical formulation. In an exemplary embodiment, the melphalan flufenamide may be in the form of a lyophilized powder pharmaceutical preparation comprising melphalan flufenamide and sucrose. For example, sucrose may be present in a weight ratio (w / w) of sucrose to melphalan flufenamide of about 2:1 to about 100:1, about 1:2 to about 1:75, or about 1:4 to about 1:75. Preferably, sucrose is present in a weight ratio (w / w) of sucrose to melphalan flufenamide of about 20:1 to about 75:1 (e.g., about 30:1 to about 70:1, about 40:1 to about 60:1, about 45:1 to about 55:1, or about 50:1). In another exemplary embodiment, the melphalan flufenamide may be in the form of a liquid pharmaceutical preparation comprising melphalan flufenamide or a salt thereof and a pharma- ceutically acceptable organic solvent (typically, the pharma- ceutically acceptable organic solvent is selected from the group consisting of propylene glycol, polyethylene glycol (PEG), and mixtures thereof; in certain embodiments, the pharmaceutical preparation consists essentially of melphalan flufenamide or a salt thereof, and propylene glycol).

[0092] In embodiments providing an injection or infusion solution comprising melphalan flufenamide, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), one or more physiologically acceptable solvents or diluents, and optionally one or more further excipients, the injection or infusion solution preferably comprises γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 1:1 to about 100:1, for example, a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) of about 1:1 to about 50:1, or about 3:1 to about 50:1. In one embodiment, the injection or infusion solution comprises γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 4:1 to about 40:1. In an exemplary embodiment, the injection or infusion solution comprises γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 4:1 to about 30:1, preferably about 4:1 to about 25:1. Thus, in certain preferred embodiments, the injection or infusion solution comprises γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 10:1 to about 25:1. For example, the injection or infusion solution comprises γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, or about 25:1.More preferably, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 13:1 to about 25:1. In one preferred embodiment, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 15:1 to about 23:1. For example, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) is about 15:1, about 18:1, or about 23:1. In another preferred embodiment, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 20:1 to about 25:1. For example, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 20:1, about 22:1, about 24:1, or about 25:1.

[0093] In certain other preferred embodiments, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 15:1 to about 25:1. For example, the injection or infusion solution contains γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, or about 25:1. In one preferred embodiment, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 16:1 to about 25:1. In a more preferred embodiment, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (e.g., HPγCD) to melphalan flufenamide is about 20: 1 to about 25: 1. For example, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (e.g., HPγCD) to melphalan flufenamide is about 20: 1, about 22: 1, about 24: 1, or about 25: 1.

[0094] Additional Therapeutic Agents Melphalan flufenamide or its salt may be present in the pharmaceutical preparation of the present invention as the only active ingredient, but it is also possible that the pharmaceutical preparation of the present invention further comprises one or more additional therapeutic agents. Such agents are known in the art. Examples of additional therapeutic agents for use in the present invention include steroids (prednisone, prednisolone, and dexamethasone), IMiDs (thalidomide, lenalidomide, and pomalidomide), PIs (bortezomib, carfilzomib, and ixazomib), histone deacetylase (HDAC) inhibitors (panobinostat), conventional chemotherapeutic agents (alkylating agents (e.g., melphalan, cyclophosphamide, bendamustine), doxorubicin), and monoclonal antibodies (e.g., anti-CD38 antibodies (daratumumab) and anti-SLAMF7 antibodies (elotuzumab)).

[0095] Thus, in certain embodiments, the pharmaceutical preparation of the invention comprises melphalan flufenamide or a salt thereof, γCD, and one or more additional therapeutic agents.

[0096] For example, the pharmaceutical preparation of the present invention may consist essentially of melphalan flufenamide or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), and one or more additional therapeutic agents. Alternatively, for example, the pharmaceutical preparation of the present invention may consist essentially of melphalan flufenamide or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), one or more additional excipients (e.g., sucrose), and one or more additional therapeutic agents. Alternatively, for example, the pharmaceutical preparation of the present invention may consist essentially of melphalan flufenamide or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), propylene glycol and / or PEG, and one or more additional therapeutic agents.

[0097] The exact amount and concentration of additional therapeutic agents that may be present in the pharmaceutical preparation or formulation or composition of the present invention may vary depending on the administration schedule, the potency of the particular agent selected, the age, size, sex and condition of the subject (typically a mammal, e.g., a human), the nature and severity of the disease or condition, and other relevant medical and physical factors.Those skilled in the art can easily determine the amount and concentration of melflufen or a salt thereof, and optionally one or more additional therapeutic agents, suitable for use according to the present invention.

[0098] treatment The pharmaceutical preparations, formulations, and compositions of the invention find use as medicines.

[0099] In certain embodiments, the pharmaceutical preparations, formulations, and compositions of the invention find use in the treatment and / or prevention of diseases or conditions that can be treated with conventional chemotherapeutic agents, such as alkylating agents (e.g., melphalan, cyclophosphamide, and bendamustine).

[0100] In certain embodiments, the pharmaceutical preparations, formulations, and compositions of the present invention find use in the treatment and / or prevention of cancer, reducing tumor growth, and / or killing tumor cells. Thus, the pharmaceutical preparations, formulations, and compositions of the present invention can be used for the cure and / or prolongation of survival of patients suffering from cancer disease. The pharmaceutical preparations, formulations, and compositions of the present invention find particular use in the treatment or prevention of cancer selected from the group consisting of multiple myeloma, breast cancer, lung cancer, ovarian cancer, glioblastoma, leukemia, and lymphoma.

[0101] In other embodiments, the pharmaceutical preparations, formulations, and compositions of the invention find use in the treatment and / or prevention of amyloidosis.

[0102] The amount of melphalan flufenamide required to achieve a therapeutic effect will vary depending on the specific route of administration and the characteristics of the subject under treatment, such as species, age, weight, sex, medical condition, the specific disease and its severity, and other relevant medical and physical factors. A physician of ordinary skill can easily determine and administer the effective amount of melphalan flufenamide required for the treatment or prevention of cancer.

[0103] The pharmaceutical preparations, formulations, and compositions of the present invention find utility in methods for treating a subject, comprising administering to the subject a pharma- ceutical effective amount of the pharmaceutical preparations, formulations, or compositions of the present invention.Typically, the pharmaceutical preparations of the present invention are administered after dilution in a physiologically acceptable solvent or diluent (e.g., saline solution or glucose solution).In embodiments in which the pharmaceutical preparation is a liquid formulation, the pharmaceutical preparation can be administered directly to the subject, for example, without prior dilution, for example, without prior dilution with a physiologically acceptable solvent or diluent.

[0104] Unit Dose The pharmaceutical preparations of the present invention may be provided as unit doses. Preferred unit doses are those containing the required dose of melphalan flufenamide or a salt thereof suitable for use according to the present invention.

[0105] In a preferred embodiment, the pharmaceutical preparation of the present invention may be provided in a vial containing a unit dose of melphalan flufenamide or a salt thereof. For example, the pharmaceutical preparation of the present invention may be provided in a vial containing a unit dose of 10-800 mg of melphalan flufenamide (excluding the mass of any counterion), for example, the vial may contain a unit dose of 10 mg-50 mg, 50 mg-150 mg, 150 mg-250 mg, 250 mg-350 mg, 350 mg-450 mg, 450 mg-550 mg, 550 mg-650 mg, or 650 mg-750 mg of melflufen (excluding the mass of any counterion). Preferably, the vial contains a unit dose of 5 mg to 15 mg, 15 mg to 25 mg, 25 mg to 50 mg, 50 mg to 75 mg, 75 mg to 125 mg, 125 mg to 175 mg, or 175 mg to 225 mg of melphalan flufenamide, such as 20 mg, 30 mg, 40 mg, 50 mg, 100 mg, 150 mg, and 200 mg of melphalan flufenamide (excluding the mass of any counterion). In certain embodiments, the vial contains a unit dose of 100 mg of melphalan flufenamide (excluding the mass of any counterion). In certain other embodiments, the vial contains a unit dose of 20 mg of melphalan flufenamide (excluding the mass of any counterion). In certain other embodiments, the vial contains a unit dose of 40 mg of melphalan flufenamide (excluding the mass of any counterion).

[0106] kit The present invention provides a kit suitable for preparing a pharmaceutical preparation according to the present invention. The kit of the present invention comprises melphalan flufenamide or a salt thereof, and γCD or a pharma- ceutically acceptable derivative of γ-cyclodextrin (such as HPγCD). Melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in the kit may be provided together as a mixture, or they may be provided separately, for example in separate vials that are subsequently combined before use of the kit (e.g., combined to prepare a pharmaceutical preparation, formulation, or composition of the present invention before use in the method or use of the present invention as defined herein). For example, melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in the kit may be provided separately in separate vials that are subsequently combined before use of the kit (e.g., combined to prepare a pharmaceutical preparation, formulation, or composition of the present invention before use in the method or use of the present invention as defined herein). In one preferred embodiment, the invention provides a kit comprising a first vial containing melphalan flufenamide and sucrose as a lyophilized powder (e.g., 20 mg melphalan flufenamide (21.48 mg melphalan flufenamide hydrochloride, i.e., Ex1) and 1000 mg sucrose as a lyophilized powder) and a second vial containing γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD). The vials are provided separately and are subsequently combined prior to use of the kit (e.g., combined to prepare a pharmaceutical preparation, formulation or composition of the invention prior to use in a method or use of the invention as defined herein).

[0107] In certain embodiments, the kit of the present invention further comprises one or more physiologically acceptable aqueous solvents or diluents, such as glucose solutions or saline solutions as described herein. Typically, the physiologically acceptable aqueous solvents or diluents are provided separately to melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (HPγCD) in the kit, or the physiologically acceptable aqueous solvents or diluents are provided together as a mixture with γCD or a pharma-ceutically acceptable derivative of γCD (HPγCD) in the kit, and melphalan flufenamide is provided separately, e.g., melphalan flufenamide is provided in a separate vial that is subsequently combined with a composition comprising γCD or a pharma-ceutically acceptable derivative of γCD (HPγCD) and a physiologically acceptable aqueous solvent or diluent prior to use of the kit (e.g., combined to prepare a composition of the present invention prior to use in a method or use of the present invention as defined herein).

[0108] In certain embodiments, the kit of the present invention further comprises one or more pharma- ceutically acceptable organic solvents selected from the group consisting of propylene glycol and PEG. The one or more PEG compounds are preferably one or more of the PEG compounds described herein. The one or more pharma- ceutically acceptable organic solvents in the kit may be provided together as a mixture with one or both of melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (HPγCD), or may be provided separately and combined with melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (HPγCD), for example, in separate vials that are subsequently combined before use of the kit. For example, the one or more pharma- ceutically acceptable organic solvents in the kit may be provided together as a mixture with γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), and melphalan flufenamide may be provided separately and combined with one or more pharma- ceutically acceptable organic solvents and a pharma- ceutical acceptable derivative of γCD (such as HPγCD) mixture before use of the kit. Alternatively, for example, one or more pharma- ceutically acceptable organic solvents in the kit may be provided together as a mixture with both melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD).

[0109] In one embodiment of the present invention, the kit of the present invention further comprises one or more additional therapeutic agents as described herein. The one or more additional therapeutic agents in the kit may be provided together in a mixture with melphalan flufenamide and one or both of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), or may be provided separately.

[0110] For the avoidance of doubt, melphalan flufenamide or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γCD (HPγCD), an optional physiologically acceptable aqueous solvent or diluent, an optional one or more pharma- ceutically acceptable organic solvents, and an optional one or more further therapeutic agents are present in the kit according to the invention in forms and amounts suitable for the preparation of a pharmaceutical preparation according to the invention. A person skilled in the art can readily determine the amounts of melflufen or a salt thereof, a pharma- ceutically acceptable organic solvent, and an optional one or more further therapeutic agents suitable for use according to the invention.

[0111] The present invention also provides kits suitable for preparing injection and infusion solutions containing melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD). Such kits include melphalan flufenamide or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), and one or more physiologically acceptable aqueous solvents or diluents, such as glucose or saline solutions, as described herein. Melphalan flufenamide, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), and one or more physiologically acceptable aqueous solvents or diluents may be provided as a mixture or separately. For example, melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) may be provided together as a mixture, and one or more physiologically acceptable aqueous solvents or diluents may be provided separately. Alternatively, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) and one or more physiologically acceptable aqueous solvents or diluents may be provided as a mixture (i.e., provided as a composition comprising γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) and one or more physiologically acceptable aqueous solvents or diluents) and melphalan flufenamide may be provided separately.

[0112] Preferably, the pharma- ceutically acceptable aqueous solvent is a 5% glucose solution (w / v in water) or a 0.9% sodium chloride solution (w / v in water). Preferably, such a solution is suitable for intravenous injection and / or infusion into a subject.

[0113] In such a kit, preferably, melphalan flufenamide or a salt thereof, and γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) are provided as a pharmaceutical preparation of the present invention, e.g., a lyophilized powder of the present invention or a liquid pharmaceutical formulation of the present invention. Thus, the present invention provides kits suitable for the preparation of injection and infusion solutions comprising a pharmaceutical preparation of the present invention (e.g., a lyophilized powder of the present invention or a liquid pharmaceutical formulation of the present invention) as described herein and one or more physiologically acceptable aqueous solvents or diluents, such as a glucose solution or a saline solution.

[0114] Alternatively, the invention provides kits suitable for the preparation of injection and infusion solutions comprising a pharmaceutical preparation comprising melphalan flufenamide or a salt thereof as described herein (e.g., a lyophilized pharmaceutical preparation comprising melphalan flufenamide or a salt thereof and sucrose), a composition comprising γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), and one or more physiologically acceptable aqueous solvents or diluents, such as a glucose solution or a saline solution.

[0115] For example, the present invention provides kits suitable for the preparation of injection and infusion solutions comprising a lyophilized pharmaceutical preparation comprising melphalan flufenamide or a salt thereof, and sucrose (e.g., 20 mg of melphalan flufenamide (21.48 mg of melphalan flufenamide hydrochloride, i.e., Ex1) and 1000 mg of sucrose as a lyophilized powder), a composition comprising γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), and one or more physiologically acceptable aqueous solutions or diluents (such as a glucose solution or a saline solution), as described herein.

[0116] In one embodiment of the invention, kits of the invention suitable for the preparation of injection and infusion solutions further comprise one or more additional therapeutic agents as described herein.

[0117] Preferably, the kit of the invention includes instructions, e.g., instructions instructing a user to mix a described amount of melphalan flufenamide with a described amount of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD).

[0118] Additionally or alternatively, the instructions may instruct the user to mix a described amount of the mixture of melphalan flufenamide and γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) with a described amount of one or more pharma- ceutically acceptable organic solvents and / or a described amount of one or more physiologically acceptable aqueous solvents.

[0119] Additionally, or alternatively, the instructions may instruct the user to mix a stated amount of melphalan flufenamide with a stated amount of one or more pharma- ceutically acceptable organic solvents containing γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) and / or a stated amount of one or more physiologically acceptable aqueous solvents containing γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD).

[0120] Additionally, or alternatively, the instructions may instruct the user to mix a described amount of melphalan flufenamide with a described amount of one or more pharma- ceutically acceptable organic solvents and / or a described amount of one or more physiologically acceptable aqueous solvents, and then mix γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) into the mixture of melphalan flufenamide, one or more pharma- ceutically acceptable organic solvents and / or one or more physiologically acceptable aqueous solvents.

[0121] Such instructions may also provide guidance as to storage conditions and / or administration instructions.

[0122] For the avoidance of doubt, melphalan flufenamide or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), one or more pharma- ceutically acceptable aqueous solvents or diluents, and optionally one or more additional therapeutic agents are present in the kit according to the invention in forms and amounts suitable for the preparation of a solution suitable for use as an injection or infusion solution. Those skilled in the art can readily determine the amounts of melflufen or a salt thereof, γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD), a physiologically acceptable aqueous solvent, and optionally one or more additional therapeutic agents suitable for use according to the invention.

[0123] Finally, in embodiments of the invention in which a kit is provided, the kit of the invention may comprise γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 1:1 to about 100:1, for example, a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 4:1 to about 80:1, about 4:1 to about 50:1, or about 4:1 to about 30:1, wherein the amount of melphalan flufenamide is the amount present in the kit. In an exemplary embodiment, the pharmaceutical preparation of the invention comprises γCD or a pharma- ceutical acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutical acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 4:1 to about 25:1, preferably about 5:1 to about 25:1, wherein the amount of melphalan flufenamide is the amount present in the kit.

[0124] In certain preferred embodiments, the kits of the invention contain γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 10:1 to about 25:1, wherein the amount of melphalan flufenamide is the amount present in the kit. For example, the kit of the present invention may contain γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) in a weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide of about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, or about 25:1. In one preferred embodiment, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 10:1 to about 23:1, and the amount of melphalan flufenamide is the amount present in the kit. For example, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) is about 15:1, about 18:1, or about 23:1. In another preferred embodiment, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 15:1 to about 25:1 (e.g., 20:1 to 25:1), and the amount of melphalan flufenamide is the amount present in the kit. For example, the weight ratio (w / w) of γCD or a pharma- ceutically acceptable derivative of γCD (such as HPγCD) to melphalan flufenamide is about 15:1, about 20:1, about 22:1, about 24:1, or about 25:1.

[0125] Equivalent The invention has been described broadly and generically herein. Those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary, and that the actual parameters, dimensions, materials, and / or configurations will depend on the particular application or applications in which the teachings of the invention are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Accordingly, the foregoing embodiments are presented by way of example only, and it is to be understood that, within the scope of the appended claims and their equivalents, the invention may be practiced otherwise than as specifically described and claimed. The invention is directed to each individual feature, system, article, material, kit, and / or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and / or methods is included within the scope of the invention, provided such features, systems, articles, materials, kits, and / or methods are not mutually inconsistent. In addition, each of the narrower groups and subgroups falling within the generic disclosure also form part of the invention. This includes the general description of the invention, subject to any condition or negative limitation that excludes any subject matter from the genus, whether or not the exclusion is specifically set forth herein.

[0126] Incorporation by Reference The contents of the articles, patents, and patent applications, and other documents and electronically available information mentioned or cited herein are incorporated herein by reference in their entirety to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference. Applicant reserves the right to physically incorporate into this application any and all materials and information from such articles, patents, patent applications, or other physical and electronic documents.

[0127] The following examples illustrate the invention. EXAMPLES

[0128] Example 1: The stabilizing effect of γ-cyclodextrin on melphalan flufenamide. The ability of γ-cyclodextrin (γCD) to stabilize melphalan flufenamide in aqueous solution was investigated and compared with other excipients.

[0129] In this example, melphalan flufenamide is used in its hydrochloride form (hereinafter referred to as Example compound 1-Ex1).The preparation method of Ex1 is disclosed in WO 01 / 96367 and WO 2016 / 180740, which are incorporated herein by reference.

[0130] Sample preparation: The following excipients were used: trehalose dihydrate, sucrose, polysorbate 80, α-cyclodextrin, β-cyclodextrin, propylene glycol, 2-hydroxypropyl-β-cyclodextrin, and γCD. Samples were prepared with a weight ratio (w / w) of excipient to Ex1 of 4:1 (w / w).

[0131] Stock solutions of each excipient were prepared in water (10 mg / mL). Aliquots of 400 μL (4 mg) of each stock solution were then transferred to separate clear 4 mL vials. The vials containing the stock solutions were then cooled and lyophilized using a Christ Alpha 2-4 LSC Plus instrument.

[0132] One mL of a 1 mg / mL solution of Ex1 in water / t-butanol (1:1 v / v) was added to each of the vials containing the lyophilized excipients to give a mixture of excipient to Ex1 in a weight ratio (w / w) of 4:1. The mixtures were cooled and lyophilized overnight.

[0133] One mL of 5% glucose solution (w / v in water) containing 0.08 mg / mL of 3-methoxybenzoic acid as an internal standard was added to each vial containing the lyophilized Ex1-excipient mixture. The vials were then mixed by shaking for 15 seconds and then shaken intermittently for 5 minutes. The solutions were transferred to Whatman® Mini-UniPrep® syringeless filters and filtered according to the manufacturer's instructions. The filtrates were transferred to glass vials to prevent further dissolution. These filtrates represent the starting concentration of melphalan flufenamide for each solution (i.e., t0 samples). The t0 samples were analyzed by HPLC as described below and then stored at 25°C-30°C for 22 hours.

[0134] Sample Analysis: The amount of Ex1 in solution in each sample was determined by HPLC analysis using a calibration curve for Ex1. For each excipient tested, the percentage of Ex1 remaining in solution after 18-20 h of storage was calculated relative to the amount of Ex1 present in the corresponding t0 sample.

[0135] HPLC analysis of each sample was performed using an Agilent 1100 Series Liquid Chromatography / Mass Selective Detector (MSD, single quadrupole) equipped with an electrospray interface and a UV diode array detector. The analysis was performed using an ACE 3 C8 (3.0 x 50 mm) column with a gradient of 5 to 95% acetonitrile, 0.1% TFA in water over 3 min at a flow rate of 1 mL / min. An injection volume of 2 μL was used.

[0136] result Table 1 shows the percentage of Ex1 remaining in each solution after storage at 25°C to 30°C for 22 hours.

[0137] [Table 1]

[0138] In the absence of excipients, Ex1 has a decomposition rate of approximately 8-10% per hour at room temperature (RT). As shown above, Ex1 was found to exhibit good stability in a γCD 5% glucose solution (w / v in water) with 69% of Ex1 remaining in solution after 22 hours of storage at 25°C-30°C.

[0139] Example 2: Effect of γ-cyclodextrin on the dissolution rate of melphalan flufenamide In a clinical situation, it is important that the dissolution of melphalan flufenamide is not adversely affected by any excipients present in the formulation. Therefore, the dissolution rate of melphalan flufenamide was investigated in the presence of the excipients tested in Example 1. In this example, melphalan flufenamide was used in the form of its hydrochloride salt (i.e., Ex1).

[0140] Sample preparation: Samples were prepared with a ratio of Ex1 (free base) to excipient of 1:4 (w / w). Stock solutions of each excipient were prepared in water (10 mg / mL). Aliquots of 400 μL (4 mg) of each stock solution were then transferred to separate clear 4 mL vials. The vials containing the stock solutions were then cooled and lyophilized using a Christ Alpha 2-4 LSC Plus instrument.

[0141] One mL of a 1 mg / mL solution of Ex1 in water / t-butanol (1:1 v / v) was added to each of the vials containing the lyophilized excipients to obtain a mixture of Ex1 to excipients in a ratio of 1:4 (w / w). The mixtures were cooled and lyophilized overnight.

[0142] 0.08 mg / ml of 5% glucose solution (w / v in water) containing 3-methoxybenzoic acid as an internal standard was added to each vial containing the lyophilized Ex1-excipient mixture. The vials were then mixed by shaking for 15 seconds. Approximately half of the solution was then transferred to a Whatman® Mini-UniPrep® syringeless filter and filtered according to the manufacturer's instructions. The filtrate was immediately transferred to a glass vial to prevent further dissolution.

[0143] Sample Analysis: The amount of Ex1 in solution in each sample was determined by HPLC analysis using a calibration curve for Ex1. The HPLC analysis of each sample was carried out using the same method as described in Example 1.

[0144] result: The amount of Ex1 dissolved in each sample is shown in Table 2.

[0145] [Table 2]

[0146] As shown above, Ex1 was found to exhibit the highest initial dissolution rate in solutions containing γCD, with 68% of Ex1 dissolving within the first 15 seconds of mixing. In the absence of excipients, Ex1 has a solubility of <0.4 mg / mL in aqueous solutions. Thus, the data shown above indicates that Ex1 exhibits a very favorable dissolution rate and solubility in the presence of γCD.

[0147] Example 3: Stabilizing effect of γ-cyclodextrin on melphalan flufenamide and Example Compound 2 in non-aqueous liquid formulations The effect of γCD on the stability of melphalan flufenamide and Example Compound 2 (Ex2) in a liquid formulation using propylene glycol as a solvent was investigated.

[0148] Sample preparation: Example compound 2 has the following structure:

[0149] [ka]

[0150] The preparation method of Ex2 is disclosed in WO 2020 / 079165, which is incorporated herein by reference. In this example, melphalan flufenamide was used in its hydrochloride form (i.e., Ex1) and Ex2 was used in its hydrochloride form to prepare liquid preparations of these compounds.

[0151] Stock solutions containing 16 mg / mL γCD in 0.9% sodium chloride solution (w / v in water) or 5% glucose solution (w / v in water) and liquid formulations of Ex1 (20 mg / mL free base) or Ex2 (20 mg / mL free base) in propylene glycol were prepared. The following samples were prepared using the stock solutions and liquid formulations: - 1.2 mg / mL of Ex1 (free base) or Ex2 (free base), 6% propylene glycol, and 15-16 mg / mL of γCD 5% glucose solution (w / v in water); - 0.6 mg / mL of Ex1 (free base) or Ex2 (free base), 3% propylene glycol, and 15-16 mg / mL of γCD 5% glucose solution (w / v in water); - 0.2 mg / ml Ex1 (free base) or Ex2 (free base), 1% propylene glycol, and 15-16 mg / mL γCD 5% glucose solution (w / v in water); - 1.2 mg / mL of Ex1 (free base) or Ex2 (free base), 6% propylene glycol, and 15-16 mg / mL of γCD in 0.9% sodium chloride solution (w / v in water); - 0.6 mg / mL of Ex1 (free base) or Ex2 (free base), 3% propylene glycol, and 15-16 mg / mL of γCD 0.9% sodium chloride solution (w / v in water); and - 0.2 mg / mL of Ex1 (free base) or Ex2 (free base), 1% propylene glycol, and 15-16 mg / ml of γCD 0.9% sodium chloride solution (w / v in water).

[0152] Briefly, samples containing 1.2 mg / mL of Ex1 (free base) or Ex2 (free base) were prepared by adding 60 μL of Ex1 or Ex2 liquid formulation to 940 μL of γCD stock solution, samples containing 0.6 mg / mL of Ex1 (free base) or Ex2 (free base) were prepared by adding 30 μL of Ex1 or Ex2 liquid formulation to 970 μL of γCD stock solution, and samples containing 0.2 mg / mL of Ex1 (free base) or Ex2 (free base) were prepared by adding 20 μL of Ex1 or Ex2 liquid formulation to 1980 μL of γCD stock solution.

[0153] The samples were mixed by vortexing for 10 seconds and then filtered through a 0.45 μm PTFE filter. The filtrates were analyzed by HPLC immediately after sample preparation (i.e., at t0) and then after storage at room temperature (approximately 23° C.) for 2 hours, 5 hours, 7 hours, or 18 hours.

[0154] Sample Analysis: The Ex1 (free base) or Ex2 (free base) concentrations in the t0 samples and in samples stored at room temperature for 2 hours, 5 hours, 7 hours, or 18 hours were determined by HPLC analysis using the method described in Example 4.

[0155] result Ex2 was found to exhibit poor stability in control solutions (6% propylene glycol and saline only, or 6% propylene glycol and 5% glucose only). γCD improved the stability of Ex2 when used in saline or 5% glucose solutions at weight ratios (w / w) of γCD to melphalan flufenamide of about 16:1, about 30:1, and about 80:1. Figure 1 shows the stabilizing effect of γCD on melphalan flufenamide when used at a weight ratio (w / w) of γCD to melphalan flufenamide of about 16:1. The stability enhancing properties of γCD were found to be independent of propylene glycol concentration. Ex1 and Ex2 were found to exhibit comparable stability and solubility and have comparable HPLC retention times.

[0156] Example 4: Stabilizing effect of γ-cyclodextrin on lyophilized melphalan flufenamide preparations made in aqueous solution In August 2022, the European Medicines Agency approved a melphalan flufenamide hydrochloride preparation for use in treating adults with multiple myeloma (cancer of the bone marrow) when their cancer has not responded to previous treatments (refractory). It is used in combination with dexamethasone (an anti-inflammatory drug) in adults who have received at least three previous therapies, including immunomodulators, proteasome inhibitors, and anti-CD38 antibodies, and whose disease has worsened since the last treatment. For patients who have undergone autologous stem cell transplantation (a procedure in which cells are removed from a patient's bone marrow and replaced with stem cells from the patient themselves), it can be used if the time from transplant to cancer recurrence is at least three years. The approved melphalan flufenamide preparation is sold in Europe under the name PEPAXTI® (another trade name for melphalan flufenamide hydrochloride is PEPAXTO®). A single-dose vial of the approved melphalan flufenamide preparation PEPAXTI® (hereinafter referred to as melphalan flufenamide formulation) contains 20 mg melphalan flufenamide as a lyophilized powder (21.48 mg melphalan flufenamide hydrochloride, i.e., Ex1) and 1000 mg sucrose. PEPAXTO® has an identical formulation. A series of experiments were performed to identify the optimal concentration of γCD to improve the stability of melphalan flufenamide formulations in aqueous solutions.

[0157] Sample preparation: A stock solution of 23 mg / mL γCD in 5% glucose (w / v in water) was used to prepare standard solutions containing 1 mg, 5 mg, 10 mg, 15 mg, 18 mg, 20 mg, or 23 mg / mL γCD.

[0158] Approximately 50 mg of melphalan flufenamide formulation (containing approximately 1 mg of melphalan flufenamide) was transferred into eight different 4 mL clear glass vials, followed by the addition of 1 mL of γCD standard solution. The solution was stirred for 5 minutes. The solution was then filtered through a 0.45 μm PTFE filter. The filtrate was analyzed by HPLC immediately after sample preparation (i.e., at t0), and then after storage at room temperature (approximately 23° C.) for 2 hours, 5 hours, 7 hours, or 18 hours.

[0159] Sample Analysis: HPLC analysis was performed using an Agilent 1260 series HPLC. A Poroshell 3 x 50 mm EC-C18 column was used with a gradient of 5% to 97% acetonitrile, 0.1% formic acid over 3 minutes and a flow rate of 1 mL / min. An injection volume of 0.2 μL was used. Melphalan flufenamide concentrations in each sample were determined by HPLC analysis using a calibration curve of melphalan flufenamide.

[0160] result: As shown in Figure 2, melphalan flufenamide was stabilized by solutions containing ratios (w / w) of 1 to 23 equivalents of γCD and 1 equivalent of melphalan flufenamide. Melphalan flufenamide was most stable in solutions containing 15, 18, and 23 equivalents of γCD and 1 equivalent of melphalan flufenamide (w / w).

[0161] Example 5: Stabilizing effect of γ-cyclodextrin on melphalan flufenamide injection solution Administration of melphalan flufenamide formulations to patients in the clinic is generally by intravenous infusion over a 30-minute period using a central venous access device. The concentration of melphalan flufenamide in the infusion solution is typically 0.1 mg / mL to 0.16 mg / mL. Because melphalan flufenamide degrades in aqueous solutions, the steps to prepare the infusion solution must be performed rapidly, and infusion to patients must begin within 60 minutes of the reconstitution step of the lyophilized powder. Storage of the prepared infusion solution at 2°C to 8°C allows for delayed administration, which should not exceed 6 hours of the reconstitution step. The stabilizing effect of γCD on melphalan flufenamide contained in the infusion solution was investigated and compared with the stability of melphalan flufenamide in an infusion solution without γCD.

[0162] Sample preparation: A control infusion solution without γCD was prepared according to the following method aligned with the protocol described in the United States Prescribing Information (USPI) for PEPAXTO® dated February 2021.

[0163] [Table 3] * Steps 3 to 5 must be completed within 30 minutes.

[0164] Infusion solutions containing γCD were prepared according to steps 1-5 above, except that step 2 included adding 320 mg of γCD to each vial (i.e., a 1:16 ratio w / w of melphalan flufenamide to γCD), followed by mixing the powders by vortexing. Steps 3-5 were then performed as above.

[0165] Immediately after preparation of each infusion solution, 4 mL was removed from the infusion bag and the melphalan flufenamide content was determined by HPLC analysis. These samples represent each infusion solution at the beginning of the stability study (i.e., t0 samples). Each infusion bag was then stored at 5° C. After storage at 5° C. for 1 hour, 3 hours, 5 hours, 23 hours, 27 hours, 29 hours, and 43 hours, a 4 mL sample of the infusion solution was removed from each infusion bag. The melphalan flufenamide content of each sample was determined by HPLC analysis.

[0166] Sample Analysis: The melphalan flufenamide content of each sample of the infusion solution was determined by HPLC analysis using the method described in Example 5.

[0167] result: After storage at 5°C for 24 hours, the melphalan flufenamide content of the control infusion solution (i.e., solution without γCD) was found to be reduced by about 7% relative to the melphalan flufenamide content of the t0 sample after storage at 5°C for 24 hours. After storage at 5°C for 43 hours, the melphalan flufenamide content of the control infusion solution was found to be reduced by about 10% relative to the melphalan flufenamide content of the t0 sample. In contrast, after storage at 5°C for 43 hours, the melphalan flufenamide content of the infusion solution containing γCD was found to be reduced by only about 2% to 3% relative to the melphalan flufenamide content of the t0 sample. See Figure 3.

[0168] Example 6: Stabilizing effect of γ-cyclodextrin derivatives on melphalan flufenamide injection solutions A stock solution of 16 mg / mL hydroxypropyl-γ-cyclodextrin in 5% glucose and 0.9% saline was prepared. A 20 mg / mL (free base) solution of the compound of Example 2 in propylene glycol was prepared. Hydroxypropyl-γ-cyclodextrin was obtained from Sigma-Aldrich (product number 779229) and had a degree of substitution of 0.6 (corresponding to an average number of hydroxypropyl groups per γCD molecule of about 4.7).

[0169] To a 2 mL clear vial was added the amount of each stock solution according to Table 4 below.

[0170] [Table 4]

[0171] Samples were stored at room temperature and analyzed by HPLC after 0, 2, 5, and 23 hours. Compound degradation was reported versus area under the curve at time = 0 hours. The percentage numbers in Table 5 below represent the percentage of remaining compound from Example 2.

[0172] [Table 5]

[0173] In entries 4, 5 and 6 there was precipitation of the compound.

[0174] Analytical HPLC was performed on an Agilent Series 1100 system using a Kinetex XB C18 (2.6 μm, 3.0 × 50 mm) column with 0.1% TFA in MiliQ HO / MeCN as the mobile phase (flow rate 1 mL / min; 5–97% MeCN in 3 min).

[0175] For each pair (1-7, 2-8, 3-9, 4-10, 5-11, and 6-12), hydroxypropyl-γ-cyclodextrin had more of the compound of Example 2 remaining at the end of storage, i.e., the solutions containing hydroxypropyl-γ-cyclodextrin had better stability.

[0176] Example 7: Stabilizing effect of γ-cyclodextrin derivatives on melphalan flufenamide injection solutions with different weight ratios A stock solution of 10% by weight hydroxypropyl-γ-cyclodextrin in saline was prepared.A stock solution of the compound of Example 2 (20 mg / mL, free base) in propylene glycol was prepared.

[0177] In a 4 mL clear vial, the stock solution of Example 2 (20 μL), a 10 wt% stock solution of hydroxypropyl-γ-cyclodextrin in saline, and saline were added to make different ratios of Example 2 and excipients according to the following Table 6. The concentration of Ex2 in the final solution was 0.2 mg / mL.

[0178] [Table 6]

[0179] Samples were analyzed at 0 hours, samples were stored at room temperature or in the refrigerator and analyzed after 18 hours. The area under the curve at t=0 hours versus remaining API (i.e., remaining melphalan flufenamide) was reported.

[0180] Analytical HPLC was performed on an Agilent Series 1100 system using a Kinetex XB C18 (2.6 μm, 3.0 × 50 mm) column with 0.1% TFA in MiliQ HO / MeCN as the mobile phase (flow rate 1 mL / min; 5–97% MeCN in 3 min).

[0181] The results are shown in Table 7 below. From Table 7, it can be seen that good stability was achieved at each of the weight ratios. It is particularly noteworthy that good stability was achieved at weight ratios of 1:32 to 1:40.

[0182] [Table 7]

[0183] Example 8: Stabilizing effect of γ-cyclodextrin derivatives on melphalan flufenamide injection solutions with different weight ratios and concentrations Stock solutions of 10 wt% hydroxypropyl-γ-cyclodextrin in saline and 10 wt% hydroxypropyl-γ-cyclodextrin in 5% glucose were prepared. A stock solution of Ex2 (20 mg / mL, free base) in propylene glycol was prepared.

[0184] Stock solutions of Ex2 (100 μL) and a 10 wt % stock solution of hydroxypropyl-γ-cyclodextrin in saline or a 10 wt % stock solution of hydroxypropyl-γ-cyclodextrin in 5% glucose (see "Medium" in Table 8) were added to clear vials according to Table 8. Additional 0.9% saline or 5% glucose was added to obtain the concentrations shown in Table 8.

[0185] [Table 8]

[0186] Samples were analyzed after 0 hours, samples were stored at room temperature and analyzed after the times indicated in the table below. The remaining API (i.e., remaining melphalan flufenamide) versus area under the curve at t=0 hours was reported. Analytical HPLC was performed on an Agilent Series 1100 system using a Kinetex XB C18 (2.6 μm, 3.0 × 50 mm) column with 0.1% TFA in MiliQ H2O / MeCN as the mobile phase (flow rate 1 mL / min). 5 to 97% MeCN in 3 min.

[0187] The results are shown below in Table 9. All solutions containing hydroxypropyl-γ-cyclodextrin showed good stability at all time points.

[0188] [Table 9]

[0189] Example 9. Lyophilization, reconstitution and stability of melphalan flufenamide in derivatives of γ-cyclodextrin at injection concentrations To a mixture of Ex1 (27.1 mg, 26.0 mg free base) and hydroxypropyl-γ-cyclodextrin (1750 mg), t-BuOH (2000 μL) and water (3500 μL) were added. Hydroxypropyl-γ-cyclodextrin was obtained from Sigma-Aldrich (product number 779229) and had a degree of substitution of 0.6 (corresponding to an average number of hydroxypropyl groups per γCD molecule of approximately 4.7). The weight ratio (w / w) of melphalan flufenamide (free base):hydroxypropyl-γ-cyclodextrin was 1:67. The mixture was stirred at room temperature for 30 minutes and then lyophilized overnight. A white fluffy lyophilized cake was formed. The lyophilized material was kept refrigerated for 5 months. 0.9% saline (9000 μL) was added to the lyophilized cake and the mixture was vortexed for 1 minute. The material was completely dissolved to make a 3.0 mg / mL solution. HPLC analysis showed a purity of >99%. The reconstituted solution was diluted to 0.10 mg / mL in 0.9% saline. The stability of the solution was analyzed after 17 hours at room temperature with 91% Ex1 remaining. The stability of the approved formulation PEPAXTI® at 0.10 mg / mL in 0.9% saline after 17 hours at room temperature is approximately 56%.

[0190] This example shows that when Ex1 was lyophilized with hydroxypropyl-γ-cyclodextrin, a high reconstituted concentration could be obtained (3 mg / mL; the reconstituted concentration of the approved formulation PEPAXTI® is 0.5 mg / mL) and its good stability was achieved after reconstitution with 0.9% saline to infusion concentrations.

[0191] Example 10. Lyophilization, reconstitution and stability of melphalan flufenamide in different ratios with derivatives of γ-cyclodextrin at injection concentrations A 10 wt% solution of hydroxypropyl-γ-cyclodextrin (1750 mg) in t-BuOH / water 1:1 (15.75 g) and a 1:1 tBuOH / water solution were prepared. Hydroxypropyl-γ-cyclodextrin was obtained from Sigma-Aldrich (product number 779229) and had a degree of substitution of 0.6 (corresponding to an average number of hydroxypropyl groups per γCD molecule of about 4.7). Example 1 was weighed into different vials and different amounts of 10 wt% hydroxypropyl-γ-cyclodextrin in tBuOH / water 1:1 and a 1:1 tBuOH / water solution were added according to Table 10 below to make a 4.0 mg / mL solution of melphalan flufenamide (i.e., Ex1 as its free base). The solution was lyophilized for 4 days.

[0192] [Table 10]

[0193] The lyophilized materials of entries 1-4 in Table 10 were reconstituted with 0.9% saline to produce 0.5 mg / mL, 1.0 mg / mL, and 2.0 mg / mL reconstituted solutions. The lyophilized materials were completely dissolved in each reconstituted solution. A portion of the 0.5 mg / mL, 1.0 mg / mL, and 2.0 mg / mL reconstituted solutions was then diluted to 0.10 mg / mL in 0.9% saline (injection concentration), and samples of these 0.10 mg / mL and 0.5 mg / mL reconstituted solutions were analyzed by HPLC after 0, 4, and 20 hours at room temperature. The stability of the solutions is reported in Table 11 below as remaining Ex1 relative to AUC at t=0 hours. The equivalents reported in Table 11 are weight equivalents relative to melphalan flufenamide (i.e., Ex1 as its free base).

[0194] As can be seen from Table 11, good stability was achieved for the 0.5 mg / mL solution and for each of the 0.1 mg / mL injection concentration reconstituted solutions at all time points and for all w / w ratios of Ex1:HPγCD.

[0195] [Table 11]

[0196] Example 11. Reconstitution of Melphalan Flufenamide in the Form of an EMA Approved Formulation, PEPAXTI®, with a Solution of a Derivative of γ-Cyclodextrin in 5% Glucose The drug PEPAXTI® approved by the EMA is a formulation consisting of melflufen hydrochloride as a lyophilized product, Ex1 (20 mg free base) and sucrose (1000 mg) in a 50 mL vial. According to the approved protocol, the formulation should be reconstituted to 0.5 mg / mL by the addition of 5% glucose (40 mL). This solution is then further diluted to 0.10-0.16 mg / mL with 0.9% saline in an infusion bag. In this example, the formulation PEPAXTI® (melflufen hydrochloride as a lyophilized product, Ex1 (20 mg free base) and sucrose (1000 mg)) was reconstituted with a solution of a derivative of γ-cyclodextrin (hydroxypropyl-γ-cyclodextrin) in a 5% glucose solution containing different amounts of γ-cyclodextrin derivative.

[0197] 5% glucose solution and 10 wt% hydroxypropyl-γ-cyclodextrin in 5% glucose solution were prepared. Hydroxypropyl-γ-cyclodextrin was obtained from Sigma-Aldrich (product number 779229) and had a degree of substitution of 0.6 (corresponding to an average number of hydroxypropyl groups per γCD molecule of about 4.7). One vial of PEPAXTI® formulation (melflufen hydrochloride as a lyophilized product, Ex1 (20 mg free base) and sucrose (1000 mg)) was distributed into smaller vials and then reconstituted to a 0.5 mg / mL concentration solution using different amounts of 5% glucose solution and 10 wt% hydroxypropyl-γ-cyclodextrin in 5% glucose solution to obtain solutions with different amounts of HPγCD excipients according to Table 12 below. The equivalent weight reported in Table 12 is the weight equivalent weight to melphalan flufenamide (i.e., Ex1 as its free base).

[0198] [Table 12]

[0199] The 0.5 mg / mL solutions in Table 11 were then diluted with 0.9% saline to 0.16 mg / mL (320 μL + 0.9% saline (680 μL)) and 0.10 mg / mL (200 μL + 0.9% saline (800 μL)) solutions to make the injection concentration reconstituted solutions. These solutions were then analyzed by HPLC after 0, 4, and 20.5 hours at room temperature. The stability of these solutions was reported as the remaining Ex1 relative to the AUC at t=0 hours. The results are shown in Table 13 below. The equivalents reported in Table 13 are weight equivalents relative to melphalan flufenamide (i.e., Ex1 as its free base).

[0200] As can be seen from Table 13, improved stability was achieved at all time points for the HPγCD containing infusion concentration reconstitution solution compared to the approved PEPAXTI® formulation reconstitution solution (5% glucose without γCD).

[0201] [Table 13]

[0202] Example 12. Reconstitution of melphalan flufenamide in the form of an EMA approved formulation, PEPAXTI®, with a solution of a derivative of γ-cyclodextrin in 0.9% saline. As mentioned above, the drug PEPAXTI® approved by the EMA is a formulation consisting of melflufen hydrochloride as a lyophilized product, Ex1 (20 mg free base), and sucrose (1000 mg) in a 50 mL vial. According to the approved protocol, the formulation should be reconstituted to 0.5 mg / mL by the addition of 5% glucose (40 mL). This solution is then further diluted to 0.10-0.16 mg / mL with 0.9% saline in an infusion bag. In this example, the formulation PEPAXTI® (melflufen hydrochloride as a lyophilized product, Ex1 (20 mg free base) and sucrose (1000 mg)) was reconstituted with a solution of a derivative of γ-cyclodextrin (hydroxypropyl-γ-cyclodextrin) in a 0.9% saline solution with different amounts of γ-cyclodextrin derivative.

[0203] 10 wt% hydroxypropyl-γ-cyclodextrin in 0.9% saline solution and 0.9% saline solution were prepared. 5 wt% hydroxypropyl-γ-cyclodextrin solution in 0.9% saline was also prepared by diluting the previous one with 0.9% saline (1:1). Hydroxypropyl-γ-cyclodextrin was obtained from Sigma-Aldrich (product number 779229) and had a degree of substitution of 0.6 (corresponding to an average number of hydroxypropyl groups per γCD molecule of about 4.7). One vial of PEPAXTI® formulation (melflufen hydrochloride as a lyophilized product, Example 1 (20 mg free base) and sucrose (1000 mg)) was distributed into smaller vials and then reconstituted with various saline solutions to different reconstituted concentrations with different amounts of excipients according to Table 14 below. Each vial was vortexed for 1 minute to obtain a clear solution. The equivalents reported in Table 14 are weight equivalents relative to melphalan flufenamide (ie, Ex1 as its free base).

[0204] [Table 14]

[0205] The solutions in Table 14 were then diluted to 0.10 mg / mL with 0.9% saline to make the infusion concentration reconstituted solutions. These solutions were then analyzed by HPLC after 0, 4, and 20.5 hours at room temperature. The stability of these solutions was reported as the remaining Ex1 versus the AUC at t=0 hours. The results are shown in Table 15 below. The equivalents reported in Table 15 are weight equivalents relative to melphalan flufenamide (i.e., Ex1 as its free base).

[0206] As can be seen from Table 15, improved stability was achieved at all time points for the HPγCD containing infusion concentration reconstitution solution compared to the approved PEPAXTI® formulation reconstitution solution (5% glucose without γCD).

[0207] [Table 15]

[0208] Example 13. Lyophilized material with and without derivatives of sucrose and γ-cyclodextrin. A solution of Ex1 (121.7 mg, 113 mg free base) in t-BuOH / water 1:1 (22.6 mL) was prepared to make a stock solution of Ex1 at 5 mg / mL. Aliquots (400 μL, 2.00 mg Ex1 as free base) were distributed into tubes. A 10% solution of sucrose in water and a 10% solution of hydroxypropyl-γ-cyclodextrin in water were prepared. Hydroxypropyl-γ-cyclodextrin was obtained from Sigma-Aldrich (product number 779229) and had a degree of substitution of 0.6 (corresponding to an average number of hydroxypropyl groups per γCD molecule of approximately 4.7). To the tubes, excipients were added according to the table below. The samples were then lyophilized for 4 days.

[0209] [Table 16]

[0210] For entry 1, the lyophilized sample was reconstituted with 5% glucose (4 mL) and the vial was vortexed for 1 minute. Entries 2 and 3 were reconstituted with 0.9% saline (1 mL) and the vial was vortexed for 1 minute. The samples were diluted to 0.10 mg / mL in 0.9% saline. These samples were then analyzed by HPLC after 0, 4, and 23 hours at room temperature. The stability of these solutions is reported in Table 17 below as remaining Ex1 relative to AUC at t=0 hours. The equivalents reported in Table 17 are weight equivalents relative to melphalan flufenamide (i.e., Ex1 as its free base). The best stability at all time points was observed for the solution containing hydroxypropyl-γ-cyclodextrin.

[0211] [Table 17]

Claims

1. A pharmaceutical preparation comprising melphalan flufenamide or a salt thereof and γ-cyclodextrin or a pharmaceutically acceptable derivative of γ-cyclodextrin.

2. 2. The pharmaceutical preparation according to claim 1, wherein the weight ratio (w / w) of γ-cyclodextrin or the pharmaceutically acceptable derivative of γ-cyclodextrin and melphalan flufenamide is 1:1 to 100:1, 4:1 to 80:1, 5:1 to 25:1, or 10:1 to 25:

1.

3. 10. The pharmaceutical preparation of claim 1, wherein the pharmaceutical preparation is a lyophilized powder.

4. 4. The pharmaceutical preparation of claim 3, wherein the pharmaceutical preparation comprises 1 mg to 30 mg of melphalan flufenamide (excluding the mass of any counterion).

5. The pharmaceutical preparation of claim 4, which comprises sucrose.

6. 6. The pharmaceutical preparation of claim 5, wherein the weight ratio (w / w) of sucrose to melphalan flufenamide is from 25:1 to 75:1, for example 50:

1.

7. 10. The pharmaceutical preparation of claim 1, which is substantially free of organic solvents.

8. 10. The pharmaceutical preparation of claim 1, wherein the pharmaceutical preparation is a liquid formulation comprising propylene glycol.

9. 9. The pharmaceutical preparation of claim 8, wherein the melphalan flufenamide or salt thereof is at a concentration of about 10 mg / mL to about 50 mg / mL (excluding the mass of any counterion).

10. 10. The pharmaceutical preparation of claim 9, wherein the melphalan flufenamide or a salt thereof is at a concentration of about 15 mg / mL to about 25 mg / mL (excluding the mass of any counterion).

11. 2. The pharmaceutical preparation of claim 1, comprising one or more additional therapeutic agents selected from the group consisting of protease inhibitors (PIs), immunomodulatory drugs (IMiDs), alkylating agents, and monoclonal antibodies (e.g., anti-CD38 antibodies or anti-SLAMF7 antibodies).

12. 2. The pharmaceutical preparation of claim 1, wherein the pharmaceutical preparation consists essentially of melphalan flufenamide or a salt thereof, γ-cyclodextrin or a pharmaceutically acceptable derivative of γ-cyclodextrin, and sucrose.

13. 2. The pharmaceutical preparation according to claim 1, wherein the pharmaceutical preparation consists essentially of melphalan flufenamide or a salt thereof, γ-cyclodextrin or a pharmaceutically acceptable derivative of γ-cyclodextrin, and propylene glycol and / or PEG.

14. 2. The pharmaceutical preparation of claim 1, wherein the melphalan flufenamide is in the form of its hydrochloride salt.

15. 2. The pharmaceutical preparation of claim 1, wherein the melphalan flufenamide is deuterated melphalan flufenamide, preferably the deuterated melphalan flufenamide has the following structure: 【Chemistry 1】

16. The pharmaceutically acceptable derivative of γ-cyclodextrin includes hydroxypropyl-γ-cyclodextrin, succinyl-γ-cyclodextrin, sulfobutylether-γ-cyclodextrin, γ-cyclodextrin sodium phosphate, γ-cyclodextrin sodium sulfate, 2-carboxyethyl-γ-cyclodextrin, acetylated-γ-cyclodextrin, and sugammadex, and preferably, the pharmaceutically acceptable derivative of γ-cyclodextrin is hydroxypropyl-γ-cyclodextrin.

2. The pharmaceutical preparation of claim 1.

17. The weight ratio (w / w) of the γ-cyclodextrin or the pharmaceutically acceptable derivative of γ-cyclodextrin to melphalan flufenamide is 15:1 to 25:1, for example, 20:1 to 25:

1.

2. The pharmaceutical preparation of claim 1.

18. A composition comprising the pharmaceutical preparation of claim 1 and a physiologically acceptable aqueous solvent or diluent.

19. A composition comprising melphalan flufenamide or a salt thereof, γ-cyclodextrin or a pharmaceutically acceptable derivative of γ-cyclodextrin, and a physiologically acceptable aqueous solvent.

20. 19. The composition of claim 18, wherein the physiologically acceptable aqueous solvent is a glucose solution, a saline solution, or a mixture thereof.

21. A kit comprising the pharmaceutical preparation of claim 1 and a physiologically acceptable aqueous solvent or diluent.

22. A kit comprising melphalan flufenamide or a salt thereof, a pharmaceutically acceptable derivative of γCD or γ-cyclodextrin, and optionally one or more physiologically acceptable aqueous solvents or diluents.

23. The kit of claim 21, wherein the melphalan flufenamide or a salt thereof is provided as a lyophilized preparation comprising (or consisting essentially of) melphalan flufenamide and sucrose, or as a liquid pharmaceutical formulation comprising (or consisting essentially of) melphalan flufenamide and propylene glycol and / or polyethylene glycol.

24. A pharmaceutical preparation according to any one of claims 1 to 17, or a composition according to any one of claims 18, 19 or 20, or a kit according to any one of claims 21, 22 or 23, for use as a medicament.

25. A pharmaceutical preparation according to any one of claims 1 to 17, or a composition according to any one of claims 18, 19 or 20, or a kit according to any one of claims 21, 22 or 23, for use in the treatment or prevention of a disease or condition that can be treated with conventional chemotherapeutic agents, such as alkylating agents (e.g. melphalan, cyclophosphamide and bendamustine).

26. A pharmaceutical preparation according to any one of claims 1 to 17, or a composition according to any one of claims 18, 19 or 20, or a kit according to any one of claims 21, 22 or 23 for use in the treatment or prevention of cancer or in the treatment or prevention of amyloidosis.

27. 24. The pharmaceutical preparation of any one of claims 1 to 17, or the composition of any one of claims 18, 19 or 20, or the kit of claim 21, 22 or 23, for use in the treatment or prevention of cancer, wherein the cancer is selected from the group consisting of multiple myeloma, breast cancer, lung cancer, ovarian cancer, glioblastoma, leukemia and lymphoma.

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