Pharmaceutical compositions of beta-substituted beta-amino acid derivative-based chemotherapeutic agents

Abstract

Disclosed are pharmaceutical compositions comprising a chemotherapeutic agent based on a beta-substituted beta-amino acid derivative. The pharmaceutical compositions include a beta-substituted beta-amino acid derivative and a cyclodextrin derivative. The pharmaceutical compositions are suitable for treating cancer.

Description

[0001] This application claims the benefit of PCT International Application No. PCT / CN2021 / 070782, filed on January 8, 2021, which is incorporated herein by reference in its entirety. Technical Field

[0002] This disclosure relates to pharmaceutical compositions of chemotherapeutic agents based on β-substituted β-amino acid derivatives. The pharmaceutical compositions comprise β-substituted β-amino acid derivatives and cyclodextrin derivatives. The pharmaceutical compositions are suitable for treating cancer. Background Technology

[0003] The capabilities of selective targeted chemotherapy are of immense value in clinical practice. Cancer is a leading cause of death in developed countries, affecting one in three people in their lifetime. Numerous treatment options exist, including surgery, chemotherapy, radiation therapy, immunotherapy, and monoclonal antibody therapy. Unfortunately, for many patients, treatment options are limited, and response rates remain low.

[0004] β-substituted β-amino acid derivatives can be used as LAT1 transport chemotherapeutic agents. However, some β-substituted β-amino acid derivatives are unstable in buffered aqueous solutions suitable for intravenous administration. Summary of the Invention

[0005] According to the present invention, a host-guest inclusion complex comprises:

[0006] Compound of formula (1):

[0007]

[0008] Or a pharmaceutically acceptable zwitterion, inner salt, or salt, wherein R 1 Selected from C 1-6 Alkyl and C 1-6 alkoxy groups; and

[0009] Cyclodextrin derivatives of formula (2):

[0010]

[0011] Or a pharmaceutically acceptable zwitterion, inner salt, or salt, wherein,

[0012] n is selected from 4, 5, and 6;

[0013] R 1 To R 9 Each of them is independently selected from hydrogen, C 1-8 Alkyl sulfonate, C 1-6 Alkyl and substituted C 1-6 Alkyl; and

[0014] R 1 To R 9 At least one of them is C 1-8 Alkyl disulfonate.

[0015] According to the present invention, a pharmaceutical composition comprises:

[0016] Compound of formula (1):

[0017]

[0018] Or a pharmaceutically acceptable zwitterion, inner salt, or salt, wherein R 1 Selected from C 1-6 Alkyl and C 1-6 alkoxy groups; and

[0019] Cyclodextrin derivatives of formula (2):

[0020]

[0021] Or a pharmaceutically acceptable zwitterion, inner salt, or salt, wherein,

[0022] n is selected from 4, 5, and 6;

[0023] R 1 To R 9 Each of them is independently selected from hydrogen, C 1-8 Alkyl sulfonate, C 1-6 Alkyl and substituted C 1-6 Alkyl; and

[0024] R 1 To R 9 At least one of them is C 1-8 Alkyl disulfonate.

[0025] According to the present invention, a pharmaceutical kit comprises a host-guest inclusion complex according to the present invention; and an aqueous solution.

[0026] According to the present invention, a method of treating a patient's cancer comprises administering a therapeutically effective amount of the pharmaceutical composition according to the present invention to a patient who requires such treatment. Detailed Implementation

[0027] For the purposes of the following detailed description, unless expressly stated otherwise, it should be understood that the embodiments provided by this disclosure may take various alternative forms and sequences of steps. Furthermore, except as indicated in any operational instance or otherwise, all numerical values ​​used in the specification and claims to express, for example, quantities of ingredients should be understood to be modified in all cases by the term “about.” Therefore, unless indicated to the contrary, the numerical parameters set forth in the following specification and appended claims are approximate values ​​that may vary depending on the desired characteristics obtained through the invention. At least, and without attempt to limit the application of the doctrine of equivalence to the scope of the claims, each numerical parameter should be interpreted at least according to the number of significant figures reported and by applying ordinary rounding techniques.

[0028] Although the numerical ranges and parameters illustrating the broad scope of the invention are approximate, the values ​​described in specific embodiments should be reported as accurately as possible. However, any numerical value inherently contains some error that is necessarily caused by the standard deviation present in its corresponding test measurement.

[0029] Furthermore, it should be understood that any numerical range listed herein is intended to include all subranges contained herein. For example, the range “1 to 10” is intended to include all subranges between the listed minimum value 1 and the listed maximum value 10 (and inclusive of 1 and 10), that is, the minimum value is equal to or greater than 1 and the maximum value is equal to or less than 10.

[0030] "Alkoxy" refers to the group -OR, where R is an alkyl group. Examples of alkoxy groups include methoxy, ethoxy, propoxy, and butoxy. Alkoxy groups can be, for example, C0. 1-6 Alkoxy, C 1-5 Alkoxy, C 1-4 Alkoxy, C 1-3 Alkyl, ethoxy, or methoxy.

[0031] "Alkyl" refers to a saturated branched or straight-chain monovalent hydrocarbon group derived by removing a hydrogen atom from a single carbon atom of a parent alkane. Alkyl groups can be, for example, C1646 ... 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl or C 1-3 Alkyl group. Alkyl groups can be methyl, ethyl, n-propyl, isopropyl, or tert-butyl.

[0032] "Alkidine" refers to a saturated branched or straight-chain divalent hydrocarbon group derived by removing two hydrogen atoms from one or two carbon atoms of a parent alkane. Alkidines can be, for example, C... 1-8 Alkyl, C 1-6 Alkyl, C 1-5 Alkyl, C 1-4 Alkyl or C 1-3Alkyl. Alkyl groups can be, for example, methylenediyl, ethylenediyl, n-propyldiyl, isopropyldiyl, or butyl.

[0033] "Alkyl sulfonate" refers to a compound in which one of the carbon atoms is bonded to a sulfonate group, -SO3 - X + Alkyl, wherein X + It is a relative cation. In alkyl disulfonates, the sulfonate can associate with the terminal carbon of the alkyl group. Alkyl disulfonates can be, for example, C10 ... 1-8 Alkyl sulfonate, C 1-6 Alkyl sulfonate, C 1-5 Alkyl sulfonate, C 1-4 Alkyl disulfonate or C 1-3 Alkyl sulfonates. In alkyl disulfonates, the terminal carbon atom can be substituted with a sulfonate. For example, C 1-4 Alkyl disulfonates can have the structure -CH2-SO3 - X + -CH2-CH2-SO3 - X + 、or -CH2-CH2-CH2-SO a - X + or -CH2-CH2-CH2-CH2-SO a - X + The relative cation can be, for example, Na. + and C 1-8 Alkyl sulfonate group.

[0034] "Substituted" refers to a group in which one or more hydrogen atoms are independently replaced by the same or different substituents. Each substituent can be independently selected from halogens, -OH, -CN, -CF3, -OCF3, oxo groups (=O), -NO2, C 1-6 Alkoxy, C 1-6 Alkyl, -COOR, -NR2, and -CONR2; wherein each R is independently selected from hydrogen and C. 1-6 Alkyl group. Each substituent may be independently selected from halogen, -NH2, -OH, C 1-3 Alkoxy and C 1-3 Alkyl, trifluoromethoxy, and trifluoromethyl. Each substituent may be independently selected from OH, methyl, ethyl, trifluoromethyl, methoxy, ethoxy, and trifluoromethoxy. Each substituent may be selected from C10. 1-3 Alkyl, oxo (=O), C 1-3 Alkyl, C 1-3 Alkyl and phenyl groups. Each substituent may be selected from -OH, -NH2, C. 1-3 Alkyl and C 1-3 Alkyl group.

[0035] The "average degree of substitution" (ADS) refers to the average number of substituents per cyclodextrin molecule. The concept of the average degree of substitution for cyclodextrin derivatives is described in PCT International Publication No. WO 2009 / 018069. As an example, cyclodextrin derivatives can be described using the following designations. Substituent abbreviations are accompanied by a subscript indicating the ADS of the substituent. For example, a β-cyclodextrin derived from sulfobutyl ether with an ADS of 6.5 is designated as SBE. 6.5 -β-CD, where SBE is an abbreviation for sulfobutyl ether group. As another example, β-cyclodextrins derived from both sulfobutyl ether and hydroxypropyl groups are represented as SBE. 4.2 -HP 2.5 -β-CD, wherein the ADS of the sulfobutyl ether (SBE) group is 4.2 and the ADS of the hydroxypropyl (HP) group is 2.5.

[0036] The term "compound" as disclosed herein includes any specific compound within the disclosed chemical formula. Compounds can be identified by their chemical structure and / or chemical name. Compounds are named using the ChemBioDraw Ultra 14.0.0.117 (CambridgeSoft, Cambridge, MA) nomenclature program. In the event of a conflict between chemical structure and chemical name, the chemical structure determines the identity of the compound. Compounds described herein may contain one or more stereosymmetry centers and / or double bonds, and therefore may exist in stereoisomeric forms such as double bond isomers (i.e., geometric isomers), enantiomers, diastereomers, tautomers, or lag isomers. Therefore, any chemical structure within the scope of this specification, described in whole or in part with relative configurations, encompasses all possible enantiomers and stereoisomers of the compound shown, including stereoisomeric pure forms (e.g., geometrically pure, enantiomeric pure, or diastereomeric pure) and mixtures of enantiomers and (diastereomeric) stereoisomers. Mixtures of enantiomers and (diastereoisomers) stereoisomers can be separated into their enantiomers or (diastereoisomers) stereoisomers using separation techniques or chiral synthesis techniques well known to those skilled in the art.

[0037] Compounds of formula (1) encompass compounds of formula (1a), formula (1b), formula (1c), and combinations thereof.

[0038] "Cyclodextrin derivatives" refer to cyclic oligosaccharides comprising five or more α-D-glucopyranoside units linked in a circular configuration, and containing one or more substituents in the glucopyranoside units linked at positions 2, 3 and / or 6 via γ-1,4-glycosidic bonds.

[0039] The “nominal concentration” of compound (1) refers to the concentration of compound (1) and non-compound (1) in the composition or solution.

[0040] "Patient" refers to a mammal, such as a human.

[0041] "Pharmaceutical acceptable" means approved or permitted by federal regulatory agencies or state governments, or listed in the United States Pharmacopeia or other generally recognized pharmacopoeia for use in animals and, more precisely, in humans.

[0042] A "pharmaceutically acceptable salt" is a salt of a compound that has the desired pharmacological activity of its parent compound. Such salts include acid addition salts, which are formed by an inorganic acid with one or more protonable functional groups, such as primary, secondary, or tertiary amines within the parent compound. Examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Salts can be formed from organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-en-1-carboxylic acid, glucohepanoic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, mucoconic acid, etc. Salts can be formed when one or more acidic protons present in the parent compound are replaced by a metal ion (e.g., alkali metal ions, alkaline earth metal ions, or aluminum ions, or combinations thereof) or coordinated with an organic base (e.g., ethanolamine, diethanolamine, triethanolamine, N-methylglucosamine, etc.). Pharmaceutically acceptable salts can be hydrochloride salts. Pharmaceutically acceptable salts can be sodium salts. In compounds having two or more ionizable groups, pharmaceutically acceptable salts can contain one or more opposing ions, such as disalts, for example, dihydrochloride salts. Examples of pharmaceutically acceptable salts are disclosed, for example, in Stahl and Wermuth (eds.), Handbook of Pharmaceutical Salts, Properties, Selection and Use, First Edition, Wiley-VCH, 2008.

[0043] "Pharmaceutically acceptable salts" include hydrates and other solvates, as well as salts in crystalline or amorphous forms. When a particular pharmaceutically acceptable salt is disclosed, it should be understood that the particular salt (e.g., a hydrochloride salt) is an example of a salt, and other salts can be formed using techniques known to those skilled in the art. Furthermore, those skilled in the art will be able to convert pharmaceutically acceptable salts into corresponding compounds, free bases, and / or free acids using techniques generally known in the art.

[0044] "Pharmaceutically acceptable medium" means a pharmaceutically acceptable diluent, a pharmaceutically acceptable adjuvant, a pharmaceutically acceptable excipient, a pharmaceutically acceptable carrier, or a combination of any of the foregoing, which allows administration of the compound provided by this disclosure to a patient without destroying the pharmacological activity of the compound and is non-toxic when administered in a dose sufficient to provide a therapeutically effective amount of the compound.

[0045] "Pharmaceutical composition" refers to a β-substituted β-amino acid derivative or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable mediator for administering the β-substituted β-amino acid derivative or a pharmaceutically acceptable salt thereof to a patient. Pharmaceutically acceptable mediators are known in the art.

[0046] "Cure" a disease means to eliminate the disease or symptom or the symptoms of the disease or symptom.

[0047] "Disease" refers to a disease, symptom, illness, or any of the aforementioned symptoms.

[0048] "Treatment" in the context of a disease or condition refers to reducing the severity of one or more clinical symptoms of a disease or condition, delaying the onset of one or more clinical symptoms of a disease or condition, and / or alleviating one or more clinical symptoms of a disease or condition.

[0049] "Treatment" of a disease or condition means to suppress a disease or condition or one or more clinical symptoms thereof, to curb the development of a disease or condition or one or more clinical symptoms thereof, to alleviate a disease or condition or one or more clinical symptoms thereof, to cause a remission of a disease or condition or one or more clinical symptoms thereof, and / or to stabilize a disease or condition or one or more clinical symptoms thereof. "Treatment" of a disease or condition or condition means to produce a clinically beneficial effect without curing the underlying disease or condition.

[0050] "Therapeutic effective amount" means the amount of a compound, such as an active pharmaceutical ingredient, sufficient to affect the treatment of a disease or at least one clinical symptom of a disease when administered to a patient. "Therapeutic effective amount" can vary depending on, for example, the following: the compound; the disease and / or the symptoms of the disease; the severity of the disease and / or the symptoms of the disease or condition; the age, weight, and / or health status of the patient to be treated; and the judgment of the prescribing physician. Therapeutic effective amount in any given situation can be determined by a person skilled in the art or can be determined by routine experiments.

[0051] "Therapeutic effective dose" refers to a dose that provides effective treatment for a disease or condition in a patient. Therapeutic effective doses can vary between compounds and between patients, and can depend on factors such as the patient's condition and the route of delivery. Therapeutic effective doses can be determined according to conventional pharmacological procedures known to those skilled in the art.

[0052] "Mediator" refers to a diluent, excipient, or carrier used to administer a compound to a patient. A mediator may be pharmaceutically acceptable. Pharmaceutically acceptable mediators are those known in the art.

[0053] "About" means within 5% of a specific value, such as within 5% of the stated concentration range or within 5% of the stated time range.

[0054] Pharmaceutical compositions and methods of using said pharmaceutical compositions are now described. The disclosed pharmaceutical compositions and methods of using said pharmaceutical compositions are not intended to limit the claims. Rather, the claims are intended to cover all alternatives, modifications, and equivalents.

[0055] The host-guest inclusion complexes provided in this disclosure may contain β-substituted β-amino acid derivatives and cyclodextrin derivatives. The host-guest inclusion complexes may be in lyophilized form.

[0056] The pharmaceutical compositions provided in this disclosure may comprise β-substituted β-amino acid derivatives and cyclodextrin derivatives. The pharmaceutical compositions may comprise the host-guest inclusion complexes of this disclosure reconstituted in an aqueous formulation. The pharmaceutical compositions provided in this disclosure may be aqueous solutions for intravenous injection. The pharmaceutical compositions provided in this disclosure may be suitable for the treatment of cancer.

[0057] Cyclodextrins have been used to improve the solubility and stability of chemotherapeutic agents such as melphalan. Cyclodextrins are a family of cyclic oligosaccharides composed of macrocyclic rings of glucose subunits linked by α-1,4 glycosidic bonds. Cyclodextrins are produced from starch via enzymatic conversion. They are used in the food, pharmaceutical, drug delivery, and chemical industries, as well as in agriculture and environmental engineering. Cyclodextrins comprise five or more α-D-glucopyranoside units, as in amylose (a segment of starch). Typical cyclodextrins contain multiple glucose monomers ranging from six to eight units in the ring, forming a cyclic shape. For example, α-cyclodextrin contains six glucose subunits, β-cyclodextrin contains seven glucose subunits, and γ-cyclodextrin contains eight glucose subunits.

[0058] Sulfonyl-substituted cyclodextrins are water-soluble and characterized by a hydrophilic outer surface surrounding an internal lipophilic cavity. Cyclodextrins and lipophilic therapeutic agents can form host-guest inclusion complexes that enhance the physicochemical properties of the drug.

[0059] The subject-guest inclusion complexes or pharmaceutical compositions provided in this disclosure may comprise β-substituted β-amino acid derivatives or combinations of β-substituted β-amino acid derivatives.

[0060] β-substituted β-amino acid derivatives may have the structure of formula (1) or a combination of β-substituted β-amino acid derivatives of formula (1):

[0061]

[0062] Or a pharmaceutically acceptable zwitterion, inner salt, or salt, wherein R 1 Selected from C 1-6 Alkyl and C 1-6 Alkyl group.

[0063] In the compound of formula (1), R 1 It can be C 1-6 alkyl.

[0064] In the compound of formula (1), R 1 It can be selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0065] In the compound of formula (1), R 1 It can be C 1-6 Alkyl group.

[0066] In the compound of formula (1), R 1 It can be selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

[0067] In the compound of formula (1), the carbon atom bonded by the amino group can have an (S) absolute configuration.

[0068] In the compound of formula (1), the carbon atom bonded by the amino group can have an (R) absolute configuration.

[0069] The compound of formula (1) may be a mixture having (S) and (R) enantiomers, such as a racemic mixture having a 1:1 ratio of (S) and (R) enantiomers; or a non-racemic mixture, such as a mixture having about 75% (S) enantiomers and about 25% (R) enantiomers. The compound of formula (1) may contain, for example, X% (S) enantiomers and 100%-X% (R) enantiomers, where X is from 0 to 100.

[0070] Compound of formula (1) may be 3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid (1a), or a pharmaceutically acceptable zwitterion, inner salt, or salt thereof:

[0071]

[0072] Compound of formula (1) may be (R)-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid (1b), or a pharmaceutically acceptable zwitterion, inner salt, or salt thereof:

[0073]

[0074] Compound of formula (1) may be (S)-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid (1c), or a pharmaceutically acceptable zwitterion, inner salt, or salt thereof:

[0075]

[0076] In the compound of formula (1), the pharmaceutically acceptable salt may be a monohydrochloride salt.

[0077] In the compound of formula (1), the pharmaceutically acceptable salt may be a di(hydrochloric acid) salt.

[0078] The salt form of the compound of formula (1) depends on the pH of the aqueous solution containing the compound of formula (1).

[0079] The compound of formula (1) can be a free base, a zwitterion, or an internal salt.

[0080] The enantiomer purity of the compound of formula (1) may be, for example, greater than about 90%, greater than about 95%, greater than about 98%, greater than about 99%, greater than about 99.5%, or greater than about 99.9%.

[0081] The compound of formula (1) is a substrate of LAT1 / 4F2hc (a large amino acid 1 transporter).

[0082] Methods for synthesizing compounds of formula (1) are disclosed in U.S. Patent Nos. 9,394,237 and 9,783,487, each of which is incorporated herein by reference in its entirety.

[0083] The subject-guest inclusion complexes or pharmaceutical compositions provided in this disclosure may contain cyclodextrin derivatives or combinations of cyclodextrin derivatives.

[0084] Cyclodextrin derivatives may have the structure of formula (2):

[0085]

[0086] Or a pharmaceutically acceptable zwitterion, inner salt, or salt, wherein,

[0087] n can be selected from 4, 5, and 6;

[0088] R 1 To R 9 Each of them can be independently selected from hydrogen, C 1-8 Alkyl sulfonate, C 1-6 Alkyl and substituted C 1-6 Alkyl; and

[0089] R 1 To R 9 At least one of them can be C 1-8 Alkyl disulfonate.

[0090] In the cyclodextrin derivatives of formula (2), n can be 4, 5 or 6.

[0091] In the cyclodextrin derivatives of formula (2), C 1-8 Alkyl sulfonates may be selected from, for example, sulfoethyl, sulfopropyl, 1-methyl-sulfopropyl, sulfobutyl, 1-methyl-sulfobutyl, 2-methyl-sulfobutyl, 1-methyl-sulfobutyl-3-yl, 2-ethyl-sulfobutyl, 3-ethyl-sulfobutyl, sulfopentyl, 1-sulfopentyl-3-yl, sulfohexyl, sulfoheptyl, and sulfoctyl sulfonates.

[0092] In the cyclodextrin derivatives of formula (2), C 1-8 Alkyl disulfonates may be selected from, for example, -(CH2)1-SO3 - X + -(CH2)2-SO3 - X + -(CH2)3-SO3 - X + -(CH2)4-SO3 - X+ -(CH2)5-SO3 - X + -(CH2)6-SO3 - X + -(CH2)7-SO3 - X + and -(CH2)8-SO3 - X + , where X + Optional from, for example, Li + Na + K + Mg 2+ and Ca 2+ .

[0093] C 1-6 The alkyl group can be selected from, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl.

[0094] Replaced C 1-6 The alkyl group may be selected from, for example, substituted methyl, substituted ethyl, substituted n-propyl, substituted isopropyl, substituted n-butyl, substituted isobutyl, substituted tert-butyl, substituted n-pentyl, and substituted n-hexyl.

[0095] Replaced C 1-6 Alkyl groups can be substituted with hydroxyl groups. 1-6 Alkyl groups, such as 2-hydroxypropyl, 3-hydroxypropyl, 2,3-dihydroxypropyl, 3-oxobutyl or 2-ethoxy-ethyl.

[0096] The average degree of substitution (ADS) of the cyclodextrin derivative of formula (2) can be, for example, about 4 to about 8, about 5 to about 8, about 5.5 to about 7.5, about 6 to about 7.5, about 6 to about 7 or about 6.5 to about 7.

[0097] In the cyclodextrin derivatives of formula (2), each R 1 To R 9 It can be independently selected from hydrogen and C. 1-8 Alkyl sulfonate, and the ADS may be, for example, about 4 to about 8, about 5 to about 8, about 5.5 to about 7.5, about 6 to about 7.5, about 6 to about 7 or about 6.5 to about 7.

[0098] In the cyclodextrin derivatives of formula (2), R 1 To R 9 At least one of the C atoms can be hydroxyl-substituted. 1-6 Alkyl, such as hydroxyl-substituted C3 alkyl, and the ADS may be, for example, about 1 to about 8, about 2 to about 8, about 3 to about 7, or about 4 to about 7.

[0099] In the cyclodextrin derivatives of formula (2), the (relative) cation can be selected from, for example, Li + Na + K + Mg 2+ Ca 2+ Quaternary ammonium cations, such as C 1-8 Tetraalkylammonium; and amine cations, such as C 1-6 Alkylamine, C 4-8 Cycloalkylamines, C 1-6 Alkylamines and C 4-8 Cycloalkylamines.

[0100] In the cyclodextrin derivatives of formula (2), the cation can be Na. + The cyclodextrin derivatives of formula (2) can be polysodium salts or mixtures of salts.

[0101] In the cyclodextrin derivative of formula (2), each of the (relative) cations can be, for example, sodium, and the number of sodium cations is equal to the number of sulfonate groups.

[0102] Cyclodextrin derivatives of formula (2) may have the structure of formula (2a):

[0103]

[0104] Each R can be independently selected from hydrogen and -(CH2)4-SO3. - Na + .

[0105] The ADS of the cyclodextrin derivative of formula (2a) may be, for example, about 6 to about 7.5, about 6.2 to about 7.3, or about 6.5 to about 7.1.

[0106] The cyclodextrin derivative of formula (2a) may be a sulfobutyl ether-β-cyclodextrin (SBE-β-CD) derivative with an ADS of, for example, about 6 to about 7.5, about 6.2 to about 7.3 or about 6.5 to about 7.1.

[0107] The cyclodextrin derivative of formula (2a) may be an SBE-β-CD derivative with an ADS of, for example, about 6.0, about 6.1, about 6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8, about 6.9 or about 7.0.

[0108] Examples of suitable sulfobutyl ether-β-cyclodextrin derivatives of formula (2a) with an ADS of about 6 to about 7 can be, for example, Purchased. Cyclodextrin is a polyanionic β-cyclodextrin derivative in which the sodium sulfonate salt is separated from the lipophilic cyclodextrin cavity by a butyl ether spacer group. Cyclodextrin derivatives are available from CyDex Pharmaceuticals and Ligand Pharmaceuticals. Cyclodextrin derivatives are also available from CycloLab Research and Development Laboratory Ltd.

[0109] The cyclodextrin derivatives of formula (2) may contain SBE. 6.5 -β-CD.

[0110] The host-guest inclusion complexes provided in this disclosure may include β-substituted β-amino acid derivatives of formula (1) and cyclodextrin derivatives of formula (2).

[0111] The host-guest inclusion complex may comprise a compound of formula (1) and a cyclodextrin of formula (2) in the following weight ratios: for example, about 1:1 to about 1:100, about 1:10 to about 1:90, about 1:20 to about 1:80, about 1:30 to about 1:70, or about 1:40 to about 1:60. The host-guest inclusion complex may comprise a compound of formula (1) and a cyclodextrin of formula (2) in the following weight ratios: for example, about 1:48 to about 1:60, about 1:50 to about 1:58, or about 1:52 to about 1:56.

[0112] The host-guest inclusion complex may comprise a compound of formula (1) and a cyclodextrin derivative of formula (2) in the following molar ratios: for example, about 1:6 to about 1:11, about 1:7 to about 1:10, about 1:7.5 to about 1:9.5, about 1:8 to about 1:9 or about 1:8.2 to about 1:8.8.

[0113] The host-guest inclusion complex may comprise, for example, a compound of formula (1) and a cyclodextrin derivative of formula (2) in a molar ratio of about 1:7.4 to about 1:9.25.

[0114] The host-guest inclusion complex can be a lyophilized product. The lyophilized product can be prepared by dissolving the β-substituted β-amino acid derivative of formula (1) and the cyclodextrin derivative of formula (2) in water adjusted to a suitable pH and then lyophilizing the solution to obtain the corresponding lyophilized host-guest inclusion complex.

[0115] The pharmaceutical compositions provided in this disclosure may comprise host-guest inclusion complexes of a β-substituted β-amino acid derivative of formula (1) and a cyclodextrin derivative of formula (2).

[0116] The pharmaceutical compositions provided in this disclosure may comprise β-substituted β-amino acid derivatives of formula (1) and cyclodextrin derivatives of formula (2).

[0117] The pharmaceutical compositions provided in this disclosure may comprise compounds of formula (1) and formula (2) in the following mass ratios: for example, about 1:1 to about 1:100, about 1:10 to about 1:90, about 1:20 to about 1:80, about 1:30 to about 1:70, or about 1:40 to about 1:60. The pharmaceutical compositions may comprise compounds of formula (1) and formula (2) in the following weight ratios: for example, about 1:48 to about 1:60, about 1:50 to about 1:58, or about 1:52 to about 1:56. A 1:1 mass ratio means 50 mg of compound (1): 50 mg of a cyclodextrin derivative. In SBE 6.5 With an average MW of 2,163 g / mol for -β-CD, this is equivalent to 150 μmol of compound (1): 23.1 μmol of SBE. 6.5 -β-CD molar ratio. A 1:1 molar ratio is equivalent to 50 mg of compound (1): 324.5 mg of SBE. 6.5 -β-CD mass ratio or 1:6.49 mass ratio.

[0118] The pharmaceutical compositions provided in this disclosure may comprise compounds of formula (1) and formula (2) in the following molar ratios: for example, about 1:6 to about 1:11, about 1:7 to about 1:10, about 1:7.5 to about 1:9.5, about 1:8 to about 1:9, or about 1:8.2 to about 1:8.8.

[0119] The pharmaceutical composition may comprise, for example, a compound of formula (1) and a cyclodextrin derivative of formula (2) in a molar ratio of about 1:7.4 to about 1:9.25.

[0120] The pharmaceutical composition may contain an aqueous formulation.

[0121] The nominal concentration of the β-substituted β-amino acid derivative of formula (1) in the reconstituted aqueous formulation may be, for example, from about 1 mg / mL to about 9 mg / mL, from about 2 mg / mL to about 8 mg / mL, from about 3 mg / mL to about 7 mg / mL, from about 4 mg / mL to about 6 mg / mL, or from about 4.5 mg / mL to about 5.5 mg / mL.

[0122] The nominal concentration of the β-substituted β-amino acid derivative of formula (1) in the aqueous additive formulation may be, for example, from about 0.1 mg / mL to about 2.0 mg / mL, from about 0.2 mg / mL to about 1 mg / mL, from about 0.2 mg / mL to about 0.8 mg / mL, from about 0.3 mg / mL to about 0.7 mg / mL, or from about 0.4 mg / mL to about 0.6 mg / mL.

[0123] The aqueous formulation may comprise a combination of a β-substituted β-amino acid derivative of formula (1) and a cyclodextrin derivative of formula (2) suspended in a sodium chloride solution, such as a sodium chloride solution of about 0.4% w / v to about 1.5% w / v, about 0.6% w / v to about 1.3% w / v, or about 0.8% w / v to about 1.1% w / v. The aqueous solution may be a sodium chloride solution, such as a physiological saline solution of about 0.9% w / v.

[0124] The pharmaceutical compositions provided in this disclosure may comprise an aqueous solution of a compound of formula (1) at a nominal concentration of, for example, from about 3.0 mg / mL to about 7.0 mg / mL, such as from about 4.0 mg / mL to about 6.0 mg / mL, or from about 4.5 mg / mL to about 5.5 mg / mL. The pharmaceutical compositions may comprise an aqueous solution of a compound of formula (1) at a nominal concentration of about 5.0 mg / mL. The aqueous solution may be a sodium chloride solution, such as about 0.9% w / v physiological saline solution, such as about 154 mM sodium chloride solution.

[0125] For intravenous administration, the pharmaceutical composition may comprise, for example, an aqueous solution of a compound of formula (1) at a nominal concentration of about 0.3 mg / mL to about 0.7 mg / mL, such as about 0.4 mg / mL to about 0.6 mg / mL or about 0.45 mg / mL to about 0.55 mg / mL. The pharmaceutical composition may comprise an aqueous solution of a compound of formula (1) at a nominal concentration of about 0.5 mg / mL. The aqueous solution may be a sodium chloride solution, such as about 0.9% saline solution.

[0126] The pH of the aqueous formulation may be, for example, about 3 to about 7, about 3.5 to about 6.5, about 4 to about 6, or about 4 to about 5.

[0127] The pharmaceutical composition may comprise a lyophilized product. The lyophilized product may comprise, for example, a lyophilized aqueous formulation provided in this disclosure. The lyophilized product may be storage-stable and may be included in a kit, for example, containing an aqueous diluent, such that the lyophilized product can be reconstituted in the aqueous diluent to provide an aqueous formulation, such as an injectable aqueous formulation. For example, the lyophilized product provided in this disclosure may be stable at 5°C for 6 months, 12 months, or 18 months. For example, the lyophilized product provided in this disclosure may be stable at 25°C / 60% RH for 6 months, 3 months, or 6 months. For example, the lyophilized product provided in this disclosure may be light-stable as determined using the ICH photostability assay.

[0128] The pharmaceutical compositions provided in this disclosure may comprise a β-substituted β-amino acid derivative of formula (1) and a cyclodextrin derivative of formula (2) in the following mass ratios: for example, about 1:1 to about 1:100, about 1:10 to about 1:90, about 1:20 to about 1:80, about 1:30 to about 1:70, or about 1:40 to about 1:60. The pharmaceutical compositions provided in this disclosure may comprise a β-substituted β-amino acid derivative of formula (1) and a cyclodextrin derivative of formula (2) in the following mass ratios: for example, about 1:48 to about 1:60, about 1:50 to about 1:58, or about 1:52 to about 1:56.

[0129] The pharmaceutical compositions provided in this disclosure may comprise a β-substituted β-amino acid derivative of formula (1) and a cyclodextrin derivative of formula (2) in the following mass ratios: about 1:54, such as about 1:50 to about 1:58, about 1:51 to about 1:57, about 1:52 to about 1:56 or about 1:53 to about 1:55.

[0130] The pharmaceutical compositions provided in this disclosure may comprise a β-substituted β-amino acid derivative of formula (1) and a cyclodextrin derivative of formula (2) in the following molar ratios: for example, about 1:6 to about 1:11, about 1:7 to about 1:10, about 1:7.5 to about 1:9.5, about 1:8 to about 1:9 or about 1:8.2 to about 1:8.8.

[0131] The pharmaceutical compositions provided in this disclosure may contain compounds of formula (1) at nominal concentrations of, for example, from about 0.25 mg / mL to about 2.0 mg / mL, such as about 0.25 mg / mL, about 0.5 mg / mL, about 1.0 mg / mL, about 1.5 mg / mL or about 2.0 mg / mL.

[0132] The pharmaceutical compositions provided in this disclosure may contain, for example, a sodium chloride solution, such as an aqueous saline solution of about 0.9% w / v.

[0133] The pharmaceutical composition may contain one or more pharmaceutically acceptable excipients. The pharmaceutical composition may contain pharmaceutically acceptable buffers and / or pH adjusters, such as acidifiers or alkalizers. After dilution with an aqueous diluent, the pH of the pharmaceutical composition may be, for example, from about 3 to about 6, such as from about 4 to about 6, or from about 4.5 to about 5.5.

[0134] The pharmaceutical composition may contain an amount of pH adjuster sufficient to provide a diluted composition with a pH of about 4 to about 6. The pharmaceutical composition may contain an aqueous solution of NaHCO3 or NaOH as a pH adjuster. The pharmaceutical composition may contain an aqueous solution of HCl as a pH adjuster.

[0135] The pharmaceutical compositions provided in this disclosure, such as aqueous sodium chloride solutions, are stable at a temperature of about 25°C for, for example, about 2 hours, about 4 hours, or about 8 hours.

[0136] The pharmaceutical compositions provided in this disclosure, such as aqueous sodium chloride solutions, are stable at a temperature of about 0°C for, for example, about 12 hours, about 24 hours, or about 36 hours.

[0137] A stable pharmaceutical composition is a composition in which, after storage under the stated storage conditions (including, for example, a specified time and at a specified temperature), greater than about 80%, greater than about 85%, greater than about 90%, greater than about 95%, or greater than about 98% of the initial amount of a compound of formula (1) or a pharmaceutically acceptable zwitterion, inner salt, or salt thereof remains.

[0138] For example, a host-guest inclusion complex containing a certain concentration of compound (1) / cyclodextrin (such as HCl salt / SBE of compound (1c) at a concentration of about 0.125 mg / mL to about 2 mg / mL). 6.5 A 0.9% saline solution of a β-CD host-guest inclusion complex is stable for about 2 to about 12 hours at about 25°C and for about 12 to about 48 hours at about 0°C. For example, a 0.9% saline solution of a compound of formula (1) / cyclodextrin host-guest inclusion complex (such as HCl salt of compound (1c) / SBE at a concentration of about 0.125 mg / mL to about 2 mg / mL) is also acceptable. 6.5 An aqueous solution of about 0.9% of the β-CD host-guest complex is stable for about 2 hours at about 25°C and for about 12 to 48 hours at about 0°C.

[0139] For example, a host-guest inclusion complex containing a certain concentration of compound (1) / cyclodextrin (e.g., compound (1c) at a concentration of about 0.5 mg / mL to about 2 mg / mL) free base / SBE 6.5 An approximately 0.9% saline solution of the β-CD host-guest inclusion complex is stable for approximately 5 to approximately 19 hours at approximately 25°C and for approximately 31 to approximately 46 hours at approximately 0°C.

[0140] After the lyophilized product containing the compound of formula (1) and the cyclodextrin derivative of formula (2) can be reconstituted in an aqueous solution (such as about 0.9% saline solution), the aqueous solution of the drug can be suitable for intravenous administration, for example for about 60 minutes, about 90 minutes, about 120 minutes, about 180 minutes or about 240 minutes.

[0141] The pharmaceutical kit provided in this disclosure may comprise a lyophilized version of the pharmaceutical composition provided in this disclosure and an aqueous solution for reconstitution of the lyophilized version to provide a formulation suitable for intravenous administration. The kit may be used to treat cancers in patients, such as cancers of the central nervous system, brain cancer, metastatic cancer, metastatic cancer of the central nervous system, or metastatic cancer of the brain.

[0142] Lyophilized products containing a β-substituted β-amino acid derivative of formula (1) and a cyclodextrin derivative of formula (2) may be provided in glass vials fitted with butyl rubber stoppers. For example, the lyophilized product may contain about 50 mg of a β-substituted β-amino acid derivative of formula (1) and about 2,700 mg of a cyclodextrin derivative of formula (2).

[0143] The kit may optionally contain an aqueous solution, such as about 0.9% saline solution, such as about 8.0 mL to about 9.0 mL, about 8.2 mL to about 8.6 mL, or about 8.3 mL to about 8.5 mL of 0.9% saline solution. An aqueous solution, such as about 0.9% saline solution, may be added to a vial containing a lyophilized compound of formula (1) / cyclodextrin host-guest inclusion complex. After mixing the lyophilized compound and the saline solution, the vial may contain about 10 mL of an aqueous solution having a nominal concentration of the compound of formula (1) of about 5 mg / mL, such as about 3 mg / mL to about 7 mg / mL, about 3.5 mg / mL to about 6.5 mg / mL, about 4 mg / mL to about 6 mg / mL, or about 4.5 mg / mL to about 5.5 mg / mL.

[0144] The kit may contain, for example, about 1 mg to about 100 mg of lyophilized host-guest inclusion complex, about 5 mg to about 80 mg, about 10 mg to about 70 mg, or about 20 mg to about 60 mg of lyophilized host-guest inclusion complex.

[0145] Any suitable vial size can be used, such as approximately 10 mL to approximately 50 mL, approximately 10 mL to approximately 40 mL, or approximately 20 mL to approximately 30 mL. Vial sizes can be, for example, approximately 10 mL, approximately 20 mL, approximately 30 mL, approximately 40 mL, or approximately 50 mL.

[0146] This solution can be further diluted by adding an additional 0.9% saline solution or by diluting an aliquot of the sample in another vial or suitable container (such as a bag with an appropriate amount of about 0.9% saline solution) to make the final theoretical or nominal concentration of the compound of formula (1) be about 0.2 mg / mL to about 1.0 mg / mL, about 0.2 mg / mL to about 0.8 mg / mL, about 0.4 mg / mL to about 0.6 mg / mL or about 0.5 mg / mL.

[0147] The pH of the reconstituted lyophilized solution may be, for example, in the range of about 4 to about 6, about 3 to about 5, about 3.5 to about 4.5, or the pH may be about 4.0.

[0148] The reconstituted lyophilized solution may contain a compound of formula (1) in the form of a free base, an HCl salt, or a combination thereof. For example, a reconstituted lyophilized solution of formula (1) may contain more than about 0% HCl salt, more than about 20%, more than about 40%, more than about 60%, or more than about 80% or about 100% HCl salt of the compound of formula (1), wherein the percentages are relative molar amounts of the compound of formula (1). For example, about 50 mg of the compound of formula (1c) corresponds to 0.150038 mmol. For example, a reconstituted solution of formula (1) may contain from about 0% to about 100% HCl salt of the compound of formula (1), from about 10% to about 90%, from about 20% to about 80%, from about 40% to about 60%, or from about 30% to about 70% HCl salt of the compound of formula (1), wherein the percentages are relative molar amounts of the compound of formula (1).

[0149] The methods provided in this disclosure include methods for administering the pharmaceutical compositions provided in this disclosure to a patient.

[0150] The drug composition can be administered intravenously via methods such as bolus injection, intravenous infusion, limb perfusion, normothermic isolated limb perfusion, and percutaneous hepatic perfusion. Intravenous administration includes administration via injection and / or via drip lines using cannulas, central lines, or external central catheter lines.

[0151] The pharmaceutical composition can be administered via infusion for an appropriate duration.

[0152] The pharmaceutical compositions provided in this disclosure are suitable for treating cancer, including solid tumors and metastatic cancer.

[0153] The amount of the compound of formula (1) that is effective in treating cancer will depend at least in part on the nature of the disease and can be determined by standard clinical techniques known in the art. Additionally, in vitro or in vivo analyses may be used to help identify the optimal dosing range. The dosing regimen and dosing intervals can also be determined by methods known to those skilled in the art. The amount of the β-substituted β-amino acid derivative / cyclodextrin derivative administered may depend in particular on the following factors: the patient being treated, the patient's weight, potential comorbidities, the patient's clinical condition, the severity of the disease, the route of administration, and the prescribing physician's judgment.

[0154] For systemic administration, the therapeutically effective dose can first be estimated by in vitro analysis. An initial dose can also be estimated using techniques known in the art based on in vivo data (e.g., animal models). This information can be used to more accurately determine the dose suitable for humans. Those skilled in the art can optimize human administration based on animal data.

[0155] The pharmaceutical compositions provided in this disclosure can be administered, for example, once daily, twice daily, and in some embodiments at intervals more than once daily. Administration can be provided alone or in combination with other drugs, and can be continued as long as necessary for effective treatment of the disease. Administration can also be performed using continuous or semi-continuous administration over a period of time. Administration includes administering the pharmaceutical composition to mammals (such as humans) in a feeding or fasting state.

[0156] The dosage and appropriate dosing interval of the compound of formula (1) can be selected to maintain a sustained therapeutically effective concentration of the compound of formula (1) in the patient's blood.

[0157] The pharmaceutical compositions provided in this disclosure can be administered using a suitable dosing regimen. The dosing regimen can be adjusted, stopped, or extended as needed to achieve therapeutic goals.

[0158] A pharmaceutical composition comprising a compound of formula (1) may be administered to treat a patient’s cancer so as to provide a therapeutically effective concentration of the compound of formula (1) in the patient’s plasma. The therapeutically effective concentration of the compound of formula (1) in the patient’s plasma may be less than an amount that would cause unacceptable adverse effects, including adverse effects on homeostasis. The therapeutically effective concentration of the compound of formula (1) in the patient’s plasma may be an amount sufficient to treat the patient’s cancer.

[0159] The pharmaceutical compositions provided in this disclosure can be applied to treat a patient’s cancer so as to provide a therapeutically effective concentration of the compound of formula (1) for a prolonged period of time, such as at least about 0.5 hours, at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, at least about 6 hours, at least about 8 hours, at least about 10 hours, or at least about 12 hours.

[0160] The amount of compound of formula (1) applied may vary during the treatment regimen.

[0161] The methods provided in this disclosure include methods for treating a patient with cancer, the methods comprising administering a therapeutically effective amount of the pharmaceutical composition provided in this disclosure to a patient who requires such treatment.

[0162] Cancer can be cancer in the brain or the central nervous system, regardless of its origin. Cancer can be a metastatic cancer in the brain or central nervous system. Cancer can be a solid tumor originating in the brain or central nervous system.

[0163] The pharmaceutical composition can be used to treat breast cancer and metastatic breast cancer.

[0164] The pharmaceutical compositions disclosed herein can be used to treat cancers of the central nervous system.

[0165] The pharmaceutical compositions disclosed herein can be used to treat brain cancer.

[0166] Cancer can be a cancer of the central nervous system or a metastasis of cancer originating from tissues other than the central nervous system.

[0167] Cancer can be primary brain cancer or metastatic brain cancer. Cancer can be primary cancer of the central nervous system or metastatic cancer of the central nervous system.

[0168] The pharmaceutical compositions disclosed herein can be used to treat metastatic cancers, such as metastatic cancers of any origin that highly express LAT1 / 4F2hc.

[0169] The pharmaceutical compositions provided in this disclosure may be administered together with one or more compounds that are effective in treating cancers treated by compounds of formula (1) and / or causing side effects from the administration of compounds of formulas (1)-(1c).

[0170] The pharmaceutical compositions provided in this disclosure can be administered concurrently with the administration of another therapeutic agent, which may be part of the same pharmaceutical composition comprising a compound of formula (1) or in a different pharmaceutical composition comprising a compound of formula (1). The compound of formula (1) may be administered before or after the administration of the other therapeutic agent. Combination therapy may be included by alternating between the administration of the compound of formula (1) and the composition comprising the other therapeutic agent, for example, to minimize adverse drug effects associated with a particular drug. When the compound of formula (1) is administered concurrently with another therapeutic agent that may produce adverse drug effects (including, for example, toxicity), the other therapeutic agent may be administered at a dose below a threshold that would trigger an adverse drug reaction.

[0171] The pharmaceutical compositions provided in this disclosure can be administered in combination with agents known or believed to be effective in treating cancer in patients.

[0172] Example

[0173] The embodiments provided in this disclosure are further illustrated by referring to the following examples, which describe the pharmaceutical compositions and their properties provided in this disclosure. Those skilled in the art will appreciate that numerous modifications can be made to both the substances and methods without departing from the scope of this disclosure.

[0174] Example 1

[0175] Solubility of the free base of compound (1c)

[0176] Mannitol (500 mg; MW 182.17 g / mol; 2.744 mmol) was added to 10 mg (0.030 mmol) of (S)-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid (compound (1c)) (free base; MW 333.25 g / mol), and dissolved in 5 mL, 10 mL, or 15 mL of distilled water by vortexing for about 30 seconds to obtain solutions with pH values ​​from 3.8 to 4.2.

[0177] Compound (1c) was not completely dissolved in 5 mL or 10 mL of mannitol solution. Compound (1c) was completely dissolved in 15 mL of mannitol solution.

[0178] Example 2

[0179] Solubility of the free base of compound (1c)

[0180] Lactose (500 mg; MW 342.30 g / mol; 1.461 mmol) was added to 10 mg (0.030 mmol) of (S)-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid (compound (1c)) (free base) and dissolved in 5 mL, 10 mL or 15 mL of distilled water by vortexing for about 30 seconds to obtain a solution with a pH of 3.8 to 4.2.

[0181] Compound (1c) was not completely dissolved in 5 mL or 10 mL of lactose solution. Compound (1c) was completely dissolved in 15 mL of lactose solution.

[0182] Example 3

[0183] Stability of free base of compound (1c)

[0184] Stability analysis was performed in 96-well microtiter plates. (S)-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid (the free base of compound (1c)) was incubated at 37 °C in phosphate-buffered saline (PBS buffer), pH 7.4. The final concentration of compound (1c) in the reaction mixture (30 μL) was 1 μM. The percentage of remaining compound (1c) in the mixture was determined by LC / MS / MS.

[0185] At each sampling time point, 300 μL of quenching solution (50 v / v acetonitrile, 50 v / v methanol containing 1.3% (v / v) 6N HCl) and internal standards (bucetin for positive ion ESI mode; warfarin for negative ion ESI mode) were transferred to each well. The plate was sealed and centrifuged at 4,000 rpm for 15 min at 4 °C. The supernatant was transferred to a fresh plate for LC / MS / MS analysis.

[0186] All samples were tested using AB Sciex API in conjunction with the Shimadzu LC-20AD LC pump system. The instrument performed LC / MS / MS analysis. A Waters Atlantis T3 dC18 reverse-phase HPLC column (20 mm × 2.1 mm) was used to separate and analyze the samples at a flow rate of 0.4 mL / min. The mobile phase consisted of water containing 0.1% formic acid (solvent A) and 100% acetonitrile containing 0.1% formic acid (solvent B). Details of the eluent gradient used are provided in Table 1. The stability results of the tested compounds in PBS buffer are provided in Table 2.

[0187] Table 1. Details of analytical HPLC gradients.

[0188]

[0189] Table 2. Stability of the free base of compound (1c) in PBS buffer.

[0190] Time (minutes) 0 15 30 60 120 240 Remaining compounds (1c) (%) 100 78 68 56 30 10

[0191] Example 4

[0192] Stability of free base of compound (1c)

[0193] Cyclodextrin derivative (270 mg; SBE) 6.5 -β-CD, 5.5 mg (0.0165 mmol) of (S)-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid (compound (1c)) (free base) with an average MW of 2,163 g / mol was added to a 12 mL glass vial and dissolved by vortexing in 3 mL to 4 mL of double-distilled water to obtain a clear, colorless solution. The solution was filtered into a 12 mL glass vial through a 0.45 μm nylon syringe filter. The filter was washed twice with double-distilled water (approximately 0.5 mL each time) to obtain a clear, colorless solution. The filtered solution was frozen at -78 °C (dry ice / acetone bath) and lyophilized at approximately 100 mTorr for approximately 16 hours to obtain a solution consisting of 5.5 mg (0.0165 mmol) of free base of compound (1c) and 270 mg of SBE. 6.5 -β-CD, A colorless powdery solid composed of cyclodextrin derivatives / compound (1c). Several vials containing host-guest inclusion complexes of cyclodextrin derivatives / compounds (1c) were prepared according to this method.

[0194] The host-guest inclusion complex of the cyclodextrin derivative / compound (1c) was reconstituted by dissolving it in 0.86 mL of 0.9% saline solution to obtain a 1.0 mL solution having a nominal concentration of 5.5 mg / mL of the host-guest inclusion complex of the cyclodextrin derivative / compound (1c).

[0195] A 1.0 mL aliquot of the reconstitution solution was diluted with 10.0 mL of 0.9% saline solution and the pH was adjusted with about 5 μL of saturated NaHCO3 aqueous solution to obtain 11.0 mL of transparent colorless solution with a pH of 5.8 to 6.1, and the nominal concentration of compound (1c) was 0.5 mg / mL.

[0196] A 1.0 mL aliquot of the reconstitution solution was diluted with 4.5 mL of 0.9% saline solution and the pH was adjusted with about 5 μL of saturated NaHCO3 aqueous solution to obtain a 5.5 mL transparent colorless solution with a nominal concentration of 1.0 mg / mL of compound (1c) and a pH of 6.1.

[0197] A 1.0 mL aliquot of the reconstitution solution was diluted with 1.75 mL of 0.9% saline solution and the pH was adjusted with approximately 10 μL of saturated NaHCO3 aqueous solution to obtain 2.75 mL of a transparent, colorless solution with a nominal concentration of 2.0 mg / mL of compound (1c) and a pH of 6.1.

[0198] The solution was placed in a glass vial and stored at room temperature (approximately 25°C) or refrigerated (approximately 0°C), and the amount of compound (1c) was measured at intervals using analytical high-performance liquid chromatography (HPLC / UV) coupled with a UV detector.

[0199] Analytical HPLC / UV was performed using an Agilent 1200 HPLC system equipped with a G1379B degasser, a G1312A binary pump, a G1367B Hip-Als, a G1316A TCC, and a G1315B DAD. Kinetex 5μm column C18 (4.6×150mm) and An Eclipse XDB C18 column (2.1 × 150 mm) and a personal computer were used for data processing. Water (solvent A) (Arrowhead, Nestle North America) and acetonitrile (MeCN; solvent B) (EMD AX0145-1 or Aldrich) were used in analytical HPLC / UV analysis. A gradient of 439134, each containing 0.1 vol% trifluoroacetic acid (TFA) (Oakwood Chemical, 001271). The HPLC / UV method was performed at a flow rate of 1.0 mL / min over a 15-minute run time using a gradient from 5 vol% solvent B to 100 vol% solvent B, with UV detection at λ = 220 nm and λ = 254 nm. Details of the eluent gradient are provided in Table 3. The percentage of the remaining compound (1c) is based on the AUC determined at an absorption wavelength of λ = 254 nm, with linear extrapolation to time zero (set as 100% AUC). The results are provided in Table 4.

[0200] Table 3. Details of analytical HPLC gradients.

[0201]

[0202] Table 4. Stability of free base of compound (1c).

[0203]

[0204] Example 5

[0205] Stability of compound (1c) hydrochloride

[0206] A stock solution was prepared by dissolving 68.8 mg (0.206 mmol) of compound (1c) (free base) in approximately 20.0 mL of an acetonitrile / water (1:1, v / v) mixture in a clean, graduated glass cylinder (25 mL) under vortexing. 0.407 mL of a 0.5072 M HCl aqueous solution (0.206 mmol, 1.0 equivalent; Fluka, 318957) was added by gentle vortexing, and the resulting clear solution was further diluted to 25.0 mL with an acetonitrile / water (1:1, v / v) mixture under gentle vortexing to obtain a stock solution of compound (1c) as a single HCl salt with a concentration of 2.75 mg / mL. This solution was filtered into a clean 50 mL Erlenmeyer flask via a 0.45 μm nylon syringe filter. Aliquots of 2.0 mL (2 × 1.0 mL; 2 × 2.75 mg) of stock solution were rapidly pipetted into clean 12 mL scintillation vials. The aliquots were frozen at -78 °C (dry ice / acetone bath) and lyophilized at approximately 100 mTorr for approximately 16 hours to obtain test vials each containing 6.1 mg (0.0165 mmol) of compound (1c) monohydrochloride (monoHCl salt) as a colorless powder (corresponding to 5.5 mg of compound (1c)) free base.

[0207] Cyclodextrin derivative (270 mg; SBE) 6.5 -β-CD, 6.1 mg (0.0165 mmol) of (S)-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid monohydrochloride (the monoHCl salt of compound (1c)) was added to a 12 mL glass vial and dissolved by vortexing in 3 to 4 mL of double-distilled water for approximately 30 seconds to obtain a clear, colorless solution. The solution was filtered into a 12 mL glass vial through a 0.45 μm nylon syringe filter. The filter was washed twice with double-distilled water (approximately 0.5 mL each time) to obtain a clear, colorless solution with a pH of 4.4 to 4.7. The filtered solution was frozen at -78 °C (dry ice / acetone bath) and lyophilized at approximately 100 mTorr for approximately 16 hours to obtain a solution consisting of 6.1 mg (0.0165 mmol) of the monoHCl salt of compound (1c) (corresponding to 5.5 mg of compound (1c) as a free base) and 270 mg of SBE. 6.5 -β-CD, A colorless powdery solid composed of ) was prepared. Several vials containing host-guest inclusion complexes of a single HCl salt of a cyclodextrin derivative / compound (1c) were prepared according to this method.

[0208] The host-guest inclusion complex was reconstituted by dissolving the monoHCl salt host-guest complex of the cyclodextrin derivative / compound (1c) in 0.86 mL of 0.9% saline to obtain a 1.0 mL solution with a nominal concentration of 6.1 mg / mL of the monoHCl salt host-guest inclusion complex of compound (1c), which is equivalent to a nominal concentration of 5.5 mg / mL of the host-guest inclusion complex of compound (1c).

[0209] A 0.5 mL aliquot of this solution was diluted with 5.0 mL of 0.9% saline solution and the pH was adjusted with 5 μL of saturated NaHCO3 aqueous solution to obtain a 5.5 mL transparent colorless solution with a nominal concentration of 0.5 mg / mL and a pH of 5.3 to 5.5 for compound (1c).

[0210] A 1.0 mL aliquot of the reconstitution solution was diluted with 4.5 mL of 0.9% saline solution and the pH was adjusted with approximately 10 μL of saturated NaHCO3 aqueous solution to obtain a 5.5 mL transparent colorless solution with a nominal concentration of 1.0 mg / mL of compound (1c) and a pH of 5-6.

[0211] A 1.0 mL aliquot of the reconstitution solution was diluted with 1.75 mL of 0.9% saline solution and the pH was adjusted with approximately 10 μL of saturated NaHCO3 aqueous solution to obtain 2.75 mL of a transparent, colorless solution with a nominal concentration of 2.0 mg / mL of compound (1c) and a pH of 5-6.

[0212] A 0.25 mL aliquot of the reconstituted solution was diluted with 11.75 mL of 0.9% saline solution and the pH was adjusted with approximately 10 μL of saturated NaHCO3 aqueous solution to obtain 11.0 mL of a transparent, colorless solution with a nominal concentration of 0.125 mg / mL of compound (1c) and a pH greater than 6.

[0213] As described in Example 4, the solution was placed in glass vials and stored at room temperature (approximately 25°C) or refrigerated (approximately 0°C), and the amount of compound (1c) was measured at intervals using analytical high-performance liquid chromatography (HPLC / UV) coupled with a UV detector. The results are presented in Table 5.

[0214] Table 5. Stability of compound (1c) hydrochloride in 0.9% saline solution.

[0215]

[0216] Example 6

[0217] Compound (1c) containing compound (1c): SBE at different pH values 6.5 -β-CD host-guest inclusion complex regulation Stability in compounds

[0218] The presence of compound (1c): SBE at different pH values ​​was determined by analytical HPLC / UV. 6.5 Stability of formulations of β-CD host-guest inclusion complexes. Compound (1c) and SBE. 6.5 The mass ratio of -β-CD is 1:54 or 1:50.

[0219] To prepare the formulation, SBE was continuously stirred. 6.5 -β-CD was dissolved in water for injection (WFI) until a clear solution was obtained. The solution was acidified by adding 1N HCl. Compound (1c) was dissolved in acidified SBE. 6.5 A solution of compound (1c) with a nominal concentration of 5.0 mg / mL was prepared by adding water to an aqueous solution of β-CD. The pH was adjusted with 1N HCl to the values ​​shown in Table 7, corresponding to the pH of the final formulation after reconstitution with 0.9% brine. The solution was filtered through a 0.22 μm syringe filter and then lyophilized to dryness to obtain a colorless powder. The lyophilized product was reconstituted with 0.9% brine to the nominal target concentration of compound (1c) of 5.0 mg / mL.

[0220] The total AUC and the AUC of impurities with relative retention times (RRTs) at 0.42 and 1.27 were also determined by analytical HPLC / UV. The RRT at 0.42 corresponds to the single hydrolysis product of compound (1c). The identity of impurities at an RRT of 1.27 has not yet been determined.

[0221] Analytical HPLC / UV was performed on an HPLC system equipped with a UV detector and a Waters system operating at 30°C. A C18 column (4.6 × 150 mm; 5 μm) and a personal computer were used for data processing. In analytical HPLC / UV analysis, a gradient of 0.01 M potassium dihydrogen phosphate solution (KH2PO4) (solvent A) and acetonitrile (MeCN; solvent B) adjusted to pH 3.0 with phosphoric acid (H3PO4):acetonitrile = 850:150 (v / v) was used. The needle was washed with a 50% aqueous acetonitrile solution. Before injection into the HPLC / UV system (10 μL injection volume), the sample was diluted to a nominal concentration of compound (1c) of 0.5 mg / mL with 0.9 M sodium chloride (NaCl) and 0.1 N hydrochloric acid (HCl). The diluent solution was prepared by diluting 5.26 g (90.0 mmol) of sodium chloride and 850 μL of concentrated hydrochloric acid (HCl) (36–38 wt%, d 1.184, 1.00 g, 10.2 mmol) to 100 mL.

[0222] The HPLC / UV method used involved a 36-minute run time with a flow rate of 1.5 mL / min using a solvent gradient from 10% vol% to 100% vol% of solvent B, and UV detection at λ = 220 nm. Details of the eluent gradient used are provided in Table 6. The results of the HPLC / UV analysis are provided in Table 7.

[0223] Table 6. Details of analytical HPLC gradients.

[0224]

[0225]

[0226] Table 7. Compound (1c) in 0.9% saline at different pH values ​​and nominal concentrations of 5.0 mg / mL: SBE 6.5 Stability of formulations of β-CD host-guest inclusion complexes.

[0227]

[0228] Example 7

[0229] Compound (1c) contains different compounds (1c): SBE 6.5 In formulations of β-CD host-guest inclusion complexes stability

[0230] Compound (1c): SBE was determined by analytical HPLC / UV at pH 4.5 in different proportions. 6.5 Stability of formulations of β-CD host-guest inclusion complexes. Compound (1c) and SBE. 6.5 The mass ratio of -β-CD is 1:54 or 1:70.

[0231] The formulation was prepared as described with respect to Example 6, except that a solution of compound (1c) with a nominal concentration of 3.3 mg / mL was prepared and the pH of the solution was adjusted to 4.5 with 1N HCl. The solution was filtered through a 0.22 μm syringe filter and then lyophilized to dryness to obtain a colorless powder. The lyophilized product was reconstituted with 0.9% saline to a nominal target concentration of compound (1c) of 0.5 mg / mL and a pH of 4.6, and then directly analyzed by HPLC / UV. The nominal target concentration of compound (1c) of 0.5 mg / mL and the pH of 4.6 represent the concentration and pH suitable for intravenous infusion.

[0232] As described for Example 6, the total AUC of the impurities and the AUC of the impurities with relative retention times (RRTs) at 0.42 and 1.27 were also determined by analytical HPLC. The RRT at 0.42 corresponds to the single hydrolysis product of compound (1c). The identity of the impurities at an RRT of 1.27 has not been determined.

[0233] The analytical HPLC / UV instrument and analytical HPLC / UV analytical method used are the same as in Example 6. The results of the HPLC / UV analysis are provided in Table 8.

[0234] Table 8. Compound (1c) in different proportions of compound (1c): SBE 6.5 Stability of formulations of β-CD host-guest inclusion complexes.

[0235]

[0236] Example 8

[0237] Compound (1c) contains compound (1c): SBE 6.5 Formulation of β-CD and mannitol host-guest inclusion complex Stability in matter

[0238] The compound (1c):SBE containing a weight ratio of 1:50:30 6.5 Stability of formulations of -β-CD:mannitol host-guest inclusion complexes compared to compounds containing 1:50 by weight (1c):SBE 6.5 A comparison of the stability of formulations of β-CD host-guest inclusion complexes was conducted.

[0239] The formulation was prepared as described with respect to Example 6. A solution of compound (1c) with a nominal concentration of 5.0 mg / mL was prepared, and the pH of the solution was adjusted to 5.5 with 1N HCl. The solution was filtered through a 0.22 μm syringe filter and then lyophilized to dryness to obtain a colorless powder. The lyophilized product was restored to the nominal target concentration of compound (1c) of 5.0 mg / mL and pH 5.5 with 0.9% saline solution, and then diluted to a nominal concentration of 0.5 mg / mL with 0.9M NaCl / 0.1N HCl as described in Example 6. The product was then analyzed directly by HPLC / UV.

[0240] As described with respect to Examples 6 and 7, the total AUC of the impurities and the AUC of the impurities with relative retention times (RRTs) at 0.42 and 1.27 were also determined by analytical HPLC / UV at 25°C and temperatures ranging from 2°C to 8°C. The RRT at 0.42 corresponds to the single hydrolysis product of compound (1c). The identity of the impurities at an RRT of 1.27 has not been determined.

[0241] The analytical HPLC / UV instruments and analytical HPLC / UV analytical methods used were the same as in Examples 6 and 7. The results of the HPLC / UV analysis are provided in Table 9.

[0242] Table 9. Compound (1c) in compounds containing: SBE 6.5 Stability of formulations of the β-CD inclusion complex.

[0243]

[0244]

[0245] Example 9

[0246] The relative stability of compounds (1c), melphalan, and bendamustine in injectable formulations sex

[0247] The stability of the compound (1c) as an HCl salt or free base was compared with that of melphalan HCl and / or bendamustine HCl in various pharmaceutical formulations and at different concentrations and temperatures of 0°C or 25°C.

[0248] The stability of the compound in pharmaceutical formulations was determined at concentrations ranging from 0.4 mg / mL to 0.5 mg / mL, 0.9 mg / mL to 1.1 mg / mL, and 1.8 mg / mL to 2.1 mg / mL.

[0249] At different times during storage, as described in detail in Examples 4 and 5, the amount of active pharmaceutical ingredient (API) (compound (1c), melphalan, or bendamustine) in the pharmaceutical formulation was determined using HPLC / UV.

[0250] For stability measurements, four pharmaceutical compositions were included: a formulation buffered with trisodium citrate (formulation 1); based on The blend (blended mixture 2); based on The blend (blend 3); and based on The blend (blending agent 4).

[0251] 1. Ingredients.

[0252] The first formulation comprises 57.5 vol% 1,2-propanediol (1,2-PG) and 30 vol% HS 15 ( A mixture of BASF (BASF AG) / Sigma-Aldrich and 12.5% ​​vol ethanol (EtOH) was prepared, and the resulting solution was further diluted and adjusted to pH 6.8 using a pH 8.1–8.3 trisodium citrate buffer (51 mM trisodium citrate·2H2O and 73.6 mM NaCl).

[0253] The pH 8.1–8.3 trisodium citrate buffer solution was prepared as follows: 1.50 g (5.10 mmol) of trisodium citrate·2H₂O (Na₃C₆H₅O₇·2H₂O; MW 294.10 g / mol; JT Baker, 3650-01) and 0.43 g (7.36 mmol) of sodium chloride (NaCl; MW 58.44 g / mol; Sigma-Aldrich, S9888) were dissolved in double-distilled water (dd-H₂O) (Arrowhead) to a final volume of 100 mL. The solution was filtered through a 0.45 μm nylon syringe filter and the pH was adjusted to 8.1–8.3 with a saturated aqueous solution of sodium bicarbonate (NaHCO₃).

[0254] The excipient mixture was prepared from the following: 4.77 g (62.7 mmol; 4.6 mL) of 1,2-propanediol (1,2-PG; MW 76.09 g / mol; d 1.036; Acros Organics, 220870010) and 2.52 g (2.40 mL, d 1.05) melt (microwave for 15 seconds). HS 15 (BASF / Sigma-Aldrich, 42966) and 0.798 g (17.32 mmol, 1.0 mL) of ethanol (EtOH, MW 46.06 g / mol, d 0.798, KOPTEC, V1016). Warm the clear, viscous mixture to approximately 40°C (water bath) before use.

[0255] As described in Example 5, a series of 12 mL glass vials were prepared by lyophilizing aliquots of the acidified stock solution, each containing 6.1 mg (0.0318 mmol) of compound (1c) monohydrochloride (corresponding to 5.5 mg of compound (1c) as a free base).

[0256] 660 μL (6 vol%) of the pre-warmed excipient mixture was added to the monohydrochloride of compound (1c) in the vial. The monohydrochloride of compound (1c) was dissolved by gentle heating and vortexing. The viscous solution was further diluted with 10,340 μL (94 vol%) of trisodium citrate buffer (pH 8.1–8.3) to give 11.0 mL of a clear, colorless formulation containing 0.5 mg / mL of compound (1c) as the monohydrochloride at pH 6–7.

[0257] 330 μL (6 vol%) of the pre-warmed excipient mixture was added to the monohydrochloride of compound (1c) in the vial. The monohydrochloride of compound (1c) was dissolved by gentle heating and vortexing. The viscous solution was further diluted with 5,170 μL (94 vol%) of trisodium citrate buffer (pH 8.1–8.3) to give 5.5 mL of clear, colorless formulation containing 1.0 mg / mL of compound (1c) as monohydrochloride at pH 6–7.

[0258] 165 μL (6 vol%) of the pre-warmed excipient mixture was added to the monohydrochloride of compound (1c) in the vial. The monohydrochloride of compound (1c) was dissolved by gentle heating and vortexing. The viscous solution was further diluted with 2,585 μL (94 vol%) of trisodium citrate buffer (pH 8.1–8.3) to give 2.75 mL of a clear, colorless formulation containing 2.0 mg / mL of compound (1c) as the monohydrochloride at pH 6–7.

[0259] As described in Example 4, the solution was placed in a glass vial and stored at room temperature (approximately 25°C), and the amount of compound (1c) derived from the monohydrochloride salt (monoHCl salt) of compound (1c) was measured at intervals using analytical high-performance liquid chromatography (HPLC / UV) coupled with a UV detector. The results are presented in Table 10.

[0260] 2. Ingredients.

[0261] This is an FDA-approved injectable formulation of melphalan-HCl. For injection. Supplied as a sterile, non-pyrolytic, lyophilized powder. Each vial contains melphalan hydrochloride equivalent to 50 mg melphalan and 20 mg povidone (polyvinylpyrrolidone, PVP). The solid powder is reconstituted in a sterile diluent (total 10 mL) containing 0.2 g sodium citrate, 6.0 mL propylene glycol, and 0.52 mL ethanol (96%), along with water for injection. This provides a nominal 5 mg / mL melphalan solution (mixture). The dose to be administered is immediately diluted in 0.9% sodium chloride injection (USP) to a concentration not exceeding 0.45 mg / mL. For injection. Administer intravenously.

[0262] 68.8 mg (0.206 mmol) of compound (1c) (free base) and 25.0 mg of [unclear text - possibly a vortex] were [unclear text - possibly a vortex] A stock solution was prepared by dissolving C-12 (povidone; Ashland, 830796) in approximately 20.0 mL of an acetonitrile / water (1:1, v / v) mixture in a clean, graduated glass cylinder (25 mL). 0.407 mL of a 0.5072 M HCl aqueous solution (0.206 mmol, 1.0 equivalent; Fruka, 318957) was added by gentle vortexing, and the resulting clear solution was further diluted to 25.0 mL with the acetonitrile / water (1:1, v / v) mixture by gentle vortexing to obtain a single HCl salt with a concentration of 2.75 mg / mL. A stock solution of compound (1c) at a concentration of 1.0 mg / mL was prepared. This solution was filtered through a 0.45 μm nylon syringe filter into a clean 50 mL Erlenmeyer flask. A 2.0 mL aliquot of this stock solution (2 × 1.0 mL; 2 × 2.75 mg) was rapidly pipetted into a clean 12 mL scintillation vial. The aliquots were frozen at -78 °C (dry ice / acetone bath) and lyophilized at approximately 100 mTorr for approximately 16 hours to obtain test vials each containing 6.1 mg (0.0165 mmol) of compound (1c) monohydrochloride (monoHCl salt) as a colorless powder (corresponding to 5.5 mg of compound (1c) free base).

[0263] 62.5 mg (0.205 mmol) melphalan (free base; Sigma-Aldrich, M2011) and 25.0 mg melphalan were administered via vortex. A stock solution was prepared by dissolving C-12 (povidone; Ashland, 830796) in approximately 20.0 mL of an acetonitrile / water (1:1, v / v) mixture in a clean, graduated glass cylinder (25 mL). 0.404 mL of a 0.5072 M HCl aqueous solution (0.205 mmol, 1.0 equivalent; Fruka, 318957) was added by gentle vortexing, and the resulting clear solution was further diluted to 25.0 mL with the acetonitrile / water (1:1, v / v) mixture by gentle vortexing to obtain a single HCl salt with a concentration of 2.50 mg / mL. A stock solution of melphalan with a concentration of 1.0 mg / mL was prepared. This solution was filtered through a 0.45 μm nylon syringe filter into a clean 50 mL Erlenmeyer flask. A 2.0 mL aliquot of this stock solution (2 × 1.0 mL; 2 × 2.50 mg) was rapidly pipette into a clean 12 mL scintillation vial. The aliquots were frozen at -78 °C (dry ice / acetone bath) and lyophilized at approximately 100 mTorr for approximately 16 hours to obtain test vials each containing 5.6 mg (0.0164 mmol) of melphalan monohydrochloride (mono-HCl salt) as a colorless powder (corresponding to 5.0 mg of melphalan as a free base).

[0264] Prepared from the following Diluent: Dissolve 12.43 g (163.4 mmol; 12.0 mL) of 1,2-propanediol (1,2-PG; MW 76.09 g / mol; d 1.036; Anchor Organics, Inc., 220870010), 0.40 g (1.36 mmol) of trisodium citrate·2H₂O (Na₃C₆H₅O₇·2H₂O; MW 294.10 g / mol; JT Baker, 3650-01), and 0.821 g (17.82 mmol, 1.04 mL) of ethanol (EtOH, MW 46.07 g / mol, d 0.798, KOPTEC, V1016) in a 25.0 mL graduated flask to a final volume of 20.0 mL. Filter the solution through a 0.45 μm nylon syringe filter into a clean 20 mL glass vial.

[0265] Add 1.0 mL A diluent was added to the vial containing the monohydrochloride of compound (1c). The monohydrochloride of compound (1c) was dissolved by gentle shaking and vortexing. The solution was further diluted with 9.0 mL of 0.9% saline to obtain 10.0 mL of clear, colorless formulation with a pH of 5.3 to 5.7, wherein the concentration of compound (1c) as monohydrochloride was 0.55 mg / mL.

[0266] Put 0.5 mL A diluent was added to the vial containing the monohydrochloride of compound (1c). The monohydrochloride of compound (1c) was dissolved by gentle shaking and vortexing. The solution was further diluted with 4.5 mL of 0.9% saline to give 5.0 mL of clear, colorless formulation at pH 5.3, in which the concentration of compound (1c) as monohydrochloride was 1.1 mg / mL.

[0267] Put 0.25 mL A diluent was added to the vial containing the monohydrochloride of compound (1c). The monohydrochloride of compound (1c) was dissolved by gentle shaking and vortexing. The solution was further diluted with 2.25 mL of 0.9% saline to give 2.5 mL of clear, colorless formulation at pH 5.3, in which the concentration of compound (1c) as monohydrochloride was 2.2 mg / mL.

[0268] Put 1.0 mL Add the diluent to the melphalan monohydrochloride in the vial. Dissolve the melphalan monohydrochloride by gentle shaking and vortexing. Further dilute the solution with 9.0 mL of 0.9% saline to obtain 10.0 mL of clear, colorless formulation with a pH of 5.5–5.8, wherein the concentration of melphalan monohydrochloride is 0.50 mg / mL.

[0269] Put 0.5 mL Add the diluent to the melphalan monohydrochloride in the vial. Dissolve the melphalan monohydrochloride by gentle shaking and vortexing. Further dilute the solution with 4.5 mL of 0.9% saline to obtain 5.0 mL of clear, colorless formulation with a pH of 5.3–5.5, wherein the concentration of melphalan monohydrochloride is 1.0 mg / mL.

[0270] Put 0.25 mL Add the diluent to the melphalan monohydrochloride in the vial. Dissolve the melphalan monohydrochloride by gentle shaking and vortexing. Further dilute the solution with 2.25 mL of 0.9% saline to obtain 2.5 mL of clear, colorless formulation with a pH of 5.0–5.3, in which the concentration of melphalan monohydrochloride is 2.0 mg / mL.

[0271] As described in Example 4, the solution was placed in a glass vial and stored at room temperature (approximately 25°C), and the amount of compound (1c) derived from the monohydrochloride salt (monoHCl salt) or melphalan derived from the monohydrochloride salt (monoHCl salt) was measured at intervals using analytical high-performance liquid chromatography (HPLC / UV) coupled with a UV detector. The results are provided in Table 10.

[0272] 3. Blend.

[0273] It is an FDA-approved injectable formulation of melphalan hydrochloride, namely 4-[bis(2-chloroethyl)amino]-L-phenylalanine hydrochloride. Supplied as a sterile white to off-white lyophilized powder in single-dose vials for intravenous use. Each vial contains 50 mg of melphalan free base equivalent to 56 mg of melphalan hydrochloride and 2,700 mg of betacyclodextrin sulfobutyl ether sodium (SBE). 6.5 -β-CD; sulfobutyl ether β-cyclodextrin sodium (NF). Physiological saline solution (0.9% sodium chloride injection, USP) (8.6 mL, as specified) for reconstitution. This brings the total volume to 10 mL, with a nominal concentration of 5 mg / mL for melphalan. The amount of medication required to dispense the patient's dose from the vial is... The required volume is added to an appropriate volume of 0.9% sodium chloride injection (USP) until the final nominal concentration of melphalan is 0.45 mg / mL. This solution is then infused via the injection port or central venous catheter.

[0274] As described in Example 4, a series of 12 mL glass vials were prepared, each containing 5.5 mg (0.0165 mmol) of the free base of compound (1c) and 270 mg of SBE. 6.5 -β-CD, ).

[0275] The host-guest inclusion complex was restored by dissolving the cyclodextrin derivative / compound (1c) free base host-guest complex in 0.86 mL of 0.9% saline to obtain a 1.0 mL solution, wherein the nominal concentration of the compound (1c) free base in the solution was 5.5 mg / mL.

[0276] A 1.0 mL aliquot of this solution was diluted with 10.0 mL of 0.9% saline solution and the pH was adjusted with 5 μL of saturated NaHCO3 aqueous solution to obtain 11.0 mL of a clear, colorless solution with a nominal concentration of 0.50 mg / mL of compound (1c) as the free base of compound (1c) and a pH of 5.8 to 6.1.

[0277] A 1.0 mL aliquot of this solution was diluted with 4.5 mL of 0.9% saline solution and the pH was adjusted with 5 μL of saturated NaHCO3 aqueous solution to obtain a 5.5 mL transparent colorless solution with a nominal concentration of 1.0 mg / mL of compound (1c) as the free base of compound (1c) and a pH of approximately 6.1.

[0278] A 1.0 mL aliquot of this solution was diluted with 1.75 mL of 0.9% saline solution and the pH was adjusted with 5 μL of saturated NaHCO3 aqueous solution to obtain 2.75 mL of a clear, colorless solution with a nominal concentration of 2.0 mg / mL of compound (1c) as the free base of compound (1c) and a pH of approximately 6.1.

[0279] As described in Example 5, a series of 12 mL glass vials were prepared, each containing 6.1 mg (0.0165 mmol of compound (1c) monohydrochloride (corresponding to 5.5 mg of compound (1c) free base) and 270 mg of SBE. 6.5 -β-CD, ).

[0280] The host-guest inclusion complex was restored by dissolving the cyclodextrin derivative / compound (1c) monoHCl salt host-guest complex in 0.86 mL of 0.9% saline solution to obtain a 1.0 mL solution with a nominal concentration of 6.1 mg / mL of compound (1c) monoHCl salt (corresponding to a nominal concentration of 5.5 mg / mL of compound (1c) free base).

[0281] A 0.5 mL aliquot of this solution was diluted with 5.0 mL of 0.9% saline solution and the pH was adjusted with 5 μL of saturated NaHCO3 aqueous solution to obtain a 5.5 mL transparent colorless solution with a nominal concentration of 0.50 mg / mL of compound (1c) monohydrochloride and a pH of 5.3 to 5.5.

[0282] A 1.0 mL aliquot of this solution was diluted with 4.5 mL of 0.9% saline solution and the pH was adjusted with 10 μL of saturated NaHCO3 aqueous solution to obtain a 5.5 mL transparent colorless solution with a nominal concentration of 1.0 mg / mL of compound (1c) monohydrochloride and a pH of 5 to 6.

[0283] A 0.5 mL aliquot of this solution was diluted with 0.875 mL of 0.9% saline solution and the pH was adjusted with 10 μL of saturated NaHCO3 aqueous solution to obtain 1.375 mL of a clear, colorless solution derived from the monohydrochloride of compound (1c) with a nominal concentration of 2.0 mg / mL and a pH of 5 to 6.

[0284] A 0.25 mL aliquot of this solution was diluted with 10.75 mL of 0.9% saline solution and the pH was adjusted with 10 μL of saturated NaHCO3 aqueous solution to obtain 11.0 mL of a clear, colorless solution with a nominal concentration of 0.125 mg / mL of compound (1c) monohydrochloride and a pH greater than 6.

[0285] As described in Example 4, a series of 12 mL glass vials were prepared, each containing 5.0 mg (0.0164 mmol melphalan free base) and 270 mg SBE. 6.5 -β-CD, ).

[0286] The host-guest inclusion complex was reconstituted by dissolving the cyclodextrin derivative / melphalan free base host-guest complex in 0.86 mL of 0.9% saline to obtain a 1.0 mL solution with a nominal concentration of 5.0 mg / mL melphalan free base.

[0287] A 1.0 mL aliquot of this solution was diluted with 10.0 mL of 0.9% saline solution and the pH was adjusted with 5 μL of saturated NaHCO3 aqueous solution to obtain 11.0 mL of a clear, colorless solution with a nominal concentration of melphalan from the free base of melphalan of 0.45 mg / mL and a pH of 5.8 to 6.1.

[0288] As described in Example 5, a series of 12 mL glass vials were prepared, each containing 5.6 mg (0.0164 mmol melphalan monohydrochloride, corresponding to 5.0 mg melphalan free base) and 270 mg SBE. 6.5 -β-CD

[0289] The host-guest inclusion complex was reconstituted by dissolving the cyclodextrin derivative / melphalan monoHCl salt host-guest complex in 0.86 mL of 0.9% saline solution to obtain a 1.0 mL solution with a nominal concentration of melphalan monoHCl salt of 5.6 mg / mL (corresponding to a nominal concentration of 5.0 mg / mL of melphalan free base).

[0290] A 0.5 mL aliquot of this solution was diluted with 5.0 mL of 0.9% saline solution and the pH was adjusted with 7 μL of saturated NaHCO3 aqueous solution to obtain a 5.5 mL clear, colorless solution with a nominal concentration of 0.45 mg / mL of melphalan monohydrochloride and a pH of 5.8 to 6.1.

[0291] As described in Example 4, the solution was placed in a glass vial and stored at room temperature (approximately 25°C), and the amount of compound (1c) derived from the monohydrochloride salt (monoHCl salt) or melphalan derived from the monohydrochloride salt (monoHCl salt) was measured at intervals using analytical high-performance liquid chromatography (HPLC / UV) coupled with a UV detector. The results are provided in Table 10.

[0292] 4. Blending agent.

[0293] This is an FDA-approved injectable formulation of bendamustine hydrochloride, namely 5-[bis(2-chloroethyl)amino]-1-methyl-1H-benzimidazole-2-butyric acid, 4-(5-(bis(2-chloroethyl)amino)-1-methyl-1H-benzimidazole-2-yl)butyric acid monohydrochloride. For injection. It is supplied in vials containing 25 or 100 mg of bendamustine HCl as a white to off-white lyophilized powder. Each vial contains 25 mg or 100 mg of bendamustine hydrochloride and 42.5 mg or 170 mg of mannitol (USP), which is reconstituted in 0.9% sodium chloride injection (USP) or 2.5% dextrose / 0.45% sodium chloride solution injection (USP) to a final concentration of 0.2 mg / mL to 0.6 mg / mL and pH adjusted to 2.5 to 3.5.

[0294] A series of 12 mL glass vials containing 12.5 mg (0.0317 mmol) bendamustine monohydrochloride (C) were prepared by weighing commercially available compounds into vials. 16 H 21 Cl2N3O2·HCl, 394.72 g / mol; MedKoo, 200470 (corresponding to 11.3 mg of bendamustine as a free base). Add 21.3 mg (0.117 mmol) of D-mannitol (C6H4O2·HCl) to each vial containing bendamustine monohydrochloride.14 O6, MW 182.17 g / mol; Sigma-Aldrich, M1902). The mixture was dissolved in 2.0 mL of double-distilled water (dd-H2O) (arrow), and the clear, colorless solution was frozen at -78 °C (dry ice / acetone bath) and the solvent was lyophilized at about 100 mTorr over about 16 hours to give a colorless solid.

[0295] The stock solution was prepared by vortexing 85.0 mg (0.255 mmol) of compound (1c) (free base) and 170.4 mg (0.935 mmol) of D-mannitol (C6H) 14 O6 (MW 182.17 g / mol; Sigma-Aldrich, M1902) and 0.503 mL of 0.5072 M HCl aqueous solution (0.255 mmol, 1.0 equivalent) (Frukka, 318957) were dissolved in approximately 16.0 mL of a mixture of acetonitrile / water (1:1, v / v) in a clean, graduated glass cylinder (25 mL) to obtain a stock solution of compound (1c) as a single HCl salt with a concentration of 5.313 mg / mL and D-mannitol concentration of 10.65 mg / mL. This solution was filtered into a clean 25 mL Erlenmeyer flask through a 0.22 μm PTFE syringe filter. An aliquot of 2.0 mL (2 × 1.0 mL; 2 × 5.313 mg) of this stock solution was rapidly pipetted into a clean 12 mL scintillation vial. The aliquots were frozen at -78°C (dry ice / acetone bath) and freeze-dried at approximately 100 mTorr for about 16 hours to obtain test vials containing 11.8 mg (0.0319 mmol) of compound (1c) monohydrochloride (monoHCl salt) (corresponding to 10.6 mg of compound (1c) as a free base) and 21.3 mg (0.117 mmol) of D-mannitol, respectively, which were in the form of colorless powder.

[0296] The substance in the vial was dissolved in 2.5 mL of double-distilled water (dd-H2O) (Arrowhead) to obtain a clear, colorless, non-viscous stock solution containing either 4.24 mg / mL of the monohydrochloride salt of the compound (1c) or 4.54 mg / mL of the monohydrochloride salt of bendamustine.

[0297] A 1.0 mL stock solution containing 4.24 mg / mL of the monohydrochloride compound (1c) was diluted with 9.0 mL of 0.9% saline to obtain a 10.0 mL test solution with a concentration of 0.424 mg / mL of the monohydrochloride compound (1c) and a pH of approximately 4. A 1.0 mL stock solution containing 4.24 mg / mL of the monohydrochloride compound (1c) was diluted with 4.0 mL of 0.9% saline to obtain a 5.0 mL test solution with a concentration of 1.06 mg / mL of the monohydrochloride compound (1c) and a pH of 3–4. A 1.0 mL stock solution containing 4.24 mg / mL of the monohydrochloride compound (1c) was diluted with 2.0 mL of 0.9% saline to obtain a 3.0 mL test solution with a concentration of 2.12 mg / mL of the monohydrochloride compound (1c) and a pH of approximately 3.

[0298] A 1.0 mL stock solution containing 4.54 mg / mL bendamustine monohydrochloride was diluted with 9.0 mL of 0.9% saline to obtain a 10.0 mL test solution with a bendamustine monohydrochloride concentration of 0.454 mg / mL and a pH of 4–5. A 1.0 mL stock solution containing 4.54 mg / mL bendamustine monohydrochloride was diluted with 4.0 mL of 0.9% saline to obtain a 5.0 mL test solution with a bendamustine monohydrochloride concentration of 1.135 mg / mL and a pH of approximately 4–4.5. A 1.0 mL stock solution containing 4.54 mg / mL bendamustine monohydrochloride was diluted with 2.0 mL of 0.9% saline to obtain a 3.0 mL test solution with a bendamustine monohydrochloride concentration of 2.27 mg / mL and a pH of approximately 4.

[0299] As described in Example 4, the solution was placed in a glass vial and stored at room temperature (approximately 25°C) or refrigerated (approximately 0°C), and the amount of compound (1c) derived from the monohydrochloride salt (monoHCl salt) of compound (1c), or bendamustine derived from the monohydrochloride salt (monoHCl salt), was measured at intervals using analytical high-performance liquid chromatography (HPLC / UV) coupled with a UV detector. The results are provided in Table 10.

[0300] Table 10. Comparative stability of compound (1c), melphalan, and bendamustine formulations at 25°C and 0°C.

[0301]

[0302] 1 Not detected.

[0303] 2 Compound (1c), melphalan or bendamustine.

[0304] Finally, it should be understood that alternative ways of implementing the embodiments disclosed herein exist. Therefore, the embodiments of the invention are to be regarded as illustrative rather than restrictive, and the claims are not limited to the details provided in this disclosure.

Claims

1. A host-guest inclusion complex comprising: ( S )-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid or a pharmaceutically acceptable salt thereof; and sulfobutyl ether-β-cyclodextrin or its pharmaceutically acceptable salt (SBE) having an average degree of substitution (ADS) of 6 to 8. 6-8 -β-CD), The ( ) S The mass ratio of 3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid or a pharmaceutically acceptable salt thereof to the sulfobutyl ether-β-cyclodextrin or a pharmaceutically acceptable salt thereof is 1:50 to 1:

60.

2. The host-guest inclusion complex according to claim 1, wherein the sulfobutyl ether-β-cyclodextrin or a pharmaceutically acceptable salt thereof has an average degree of substitution of 6.5 to 7.

3. The host-guest inclusion complex according to claim 1, wherein the cyclodextrin derivative is a polysodium salt.

4. The host-guest inclusion complex according to claim 1, wherein ( S The molar ratio of 3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid or a pharmaceutically acceptable salt thereof to the sulfobutyl ether-β-cyclodextrin or a pharmaceutically acceptable salt thereof is 1:7 to 1:

10.

5. The host-guest inclusion complex according to claim 1, wherein ( S )-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid or a pharmaceutically acceptable salt thereof is ( S )-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyrate salt.

6. The host-guest inclusion complex according to claim 1, wherein the host-guest inclusion complex is in the form of a lyophilized product.

7. A pharmaceutical composition comprising a host-guest inclusion complex according to any one of claims 1 to 6.

8. The pharmaceutical composition according to claim 7, wherein the pharmaceutical composition comprises 1 mg / mL to 10 mg / mL of a solution dissolved in an aqueous diluent. S )-3-amino-4-(5-(bis(2-chloroethyl)amino)-2-methylphenyl)butyric acid.

9. The pharmaceutical composition of claim 8, wherein the aqueous diluent comprises a sodium chloride solution.

10. A pharmaceutical kit comprising: a host-guest inclusion complex according to any one of claims 1 to 6; and an aqueous solution.

11. The pharmaceutical kit according to claim 10, wherein the aqueous solution is a sodium chloride solution.

Images