Patent Application
20170368052
The present invention provides solid crystalline forms of apomorphine free base or a hydrate or solvate thereof, as well as a method for the preparation thereof, and liquid formulations obtained by dissolving said crystalline forms of apomorphine in a solvent. Such formulations are useful in the treatment of a neurological or movement disorder, e.g., Parkinson's disease, or a condition associated therewith.
Abbreviations: BHT, butylated hydroxytoluene; DCM, dichloromethane; DMSO, dimethyl sulfoxide; DSC, differential scanning calorimetry; DTA, differential thermal analysis; FTIR, fourier transform infrared spectroscopy; GVS, gravimetric vapour sorption; HPLC, high performance liquid chromatography; HSM, hot stage microscopy; IPA, isopropanol (isopropyl alcohol); IR, infrared spectroscopy; J, Joule; KF, Karl Fischer (determination of the water content by coulometric titration); LC-MS, liquid chromatography-mass spectrometry; MEK, methyl ethyl ketone; MET/CR, chromatography method reference; NMR, nuclear magnetic resonance; pKa, −log (Ka), acid dissociation constant; PLM, polarized light microscopy; RH, relative humidity (water activity*100); RRT, relative retention time; RT, room temperature (ambient, typically: 18 to 23° C.); STA, simulated thermal analysis (STA=TGA+DTA); TBME, tert-butyl methyl ether; TCNB, 2,3,5,6-tetrachloronitrobenzene; TGA, thermogravimetric analysis; THF, tetrahydrofuran; TMP, 2,2,6,6-tetramethylpiperidine; w/w, weight/weight; XRPD, X-ray powder diffraction.
Parkinson's disease is a progressive degenerative disease of the central nervous system. Although the primary cause of Parkinson's disease is not known, it is characterized by the degeneration of dopaminergic neurons of the substantia nigra. The substantia nigra is located in the midbrain and is involved in controlling voluntary movements. The degeneration of neurons causes a shortage of dopamine in the brain, which is believed to cause the observable symptoms of the disease. These symptoms include paucity of movement and rigidity, resting tremor, bradykinesia, and poor balance.
There are a variety of therapeutic treatments available for Parkinson's disease. The best known is levodopa, a dopamine precursor; however, treatment with levodopa can cause serious side-effects, especially over a long term. One such complication of long-term treatment with levodopa is the development of rapid fluctuations in clinical state, where a patient switches suddenly between mobility and immobility for periods ranging from a few minutes to a few hours. This phenomenon is known as the “on-off effect”, the “on” state characterized by the levodopa benefit of early normal motor functioning and the “off” state characterized by akinesia—abrupt loss of mobility, e.g., where a patient may suddenly stop while walking. Approximately half of patients on levodopa therapy will develop such on-off effects after several years of therapy.
While apomorphine hydrochloride has proved effective in treating “off” episodes in patients with Parkinson's disease, a common and serious side effect of administering apomorphine hydrochloride by subcutaneous injection is the development of subcutaneous nodules at the injection site, which can become infected, necessitating treatment or surgical involvement. A majority of people on infused apomorphine develop nodules, and a new nodule may form every time the infusion needle is re-sited, which may happen on a daily basis. Such nodules may be painful, limit available infusion sites and interfere with absorption. Further, unstable compositions (e.g., having precipitate of apomorphine or other agents) may be the cause, or exacerbate, such nodule side effects. Thus, there is a need for new, stable formulations of apomorphine which are safe and effective for administration to patients.
In one aspect, the present invention provides a solid crystalline form of apomorphine free base or a hydrate, solvate, or co-crystal thereof, more particularly, such a solid crystalline form of apomorphine free base or a solvate thereof, e.g., an alcohol solvate crystal of apomorphine free base. In a particular embodiment, the present invention provides a solid crystalline form of apomorphine solvate, wherein the solvate forming solvent is (C1-C8)alkanol, preferably IPA, i.e., to a solid crystalline form of apomorphine•IPA.
In another aspect, the present invention provides a liquid formulation obtained by dissolving a solid crystalline form of apomorphine free base or hydrate, solvate, or co-crystal thereof, as disclosed herein, e.g., said solid crystalline form of apomorphine free base or solvate thereof, in a solvent. The liquid formulation of the invention may further comprise an antioxidant. Particular such liquid formulations further comprise one or more pharmaceutically acceptable carriers or excipients, i.e., are pharmaceutically acceptable liquid formulations.
In yet another aspect, the present invention relates to a method of treating a neurological or movement disease or disorder, or a condition associated therewith, in a patient in need thereof, comprising administering to said patient a liquid formulation as disclosed herein. Examples of neurological or movement diseases or disorders include, without being limited to, Parkinson's disease, Alzheimer's disease, and akinesia, and non-limiting examples of conditions associated with neurological or movement diseases or disorders include alcoholism, opiate addiction, and erectile dysfunction.
In still another aspect, the present invention relates to a liquid formulation as disclosed herein, for use in the treatment of a neurological or movement disease or disorder, or a condition associated therewith.
In a further aspect, the present invention relates to a method of producing said solid crystalline form of apomorphine free base or a solvate thereof, said method comprising:
In particular embodiments, the method of the present invention comprises, in step (a), dissolving apomorphine hydrochloride and, optionally, said antioxidant, in IPA, to thereby obtained, following step (c), a solid crystalline form of apomorphine•IPA.
In one aspect, the present invention provides a solid crystalline form of apomorphine free base or a hydrate, solvate, or co-crystal thereof. In a more particular such aspect, the present invention provides a solid crystalline form of apomorphine free base or a solvate thereof. Such crystalline forms of apomorphine can be advantageous over amorphous form of apomorphine, e.g., amorphous salt forms such as acid addition salts of apomorphine, due to their increased/greater stability and/or improved pharmacological properties, e.g., decreased adverse reactions such as nodule side effects at the site of administration, as compared to the corresponding amorphous forms.
The term “solvate”, with respect to the solid crystalline form of the present invention, refers to a solid crystalline form consisting of apomorphine free base molecules and molecules of one or more solvents each referred to herein as “a solvate forming solvent”. Solid crystalline forms of apomorphine free base solvate comprising molecules of more than one solvent are also referred to herein as “solid crystalline form of apomorphine free base mixed solvate”. As shown herein, such crystalline forms can be prepared by crystallization from a solvent or a mixture of more than one, e.g., two or three, solvents in which the apomorphine free base is dissolved. Yet, in cases wherein crystallization is carried out from a solvent mixture, and depending on the crystallization procedure and conditions, the solid crystalline forms of the apomorphine solvate obtained may comprise molecules of one or more of the solvents present in said solvent mixture.
In certain embodiments, the present invention provides a solid crystalline form of apomorphine solvate as defined above, wherein the solvate forming solvent is selected from a (C1-C3)alkyl-, dialkyl-, or trialkylbenzene, pyridine, pyrrole, (C1-C3)alkyl-CN, (C1-C3)alkyl-NO2, (R)2NC(O)H wherein R is H or (C1-C6)alkyl, (C1-C5)alkylC(O)O— esters such as (C1-C5)alkyl-C(O)O—(C1-C5)alkyl, straight or branched (C1-C8)alkanol, i.e., (C1-C8)alcohol, (C2-C8)alkyl-O(C1-C8)alkyl, (C3-C8)cyclic ether, (C3-C7)cyclic diether, (C2-C6)glycol, or a mixture thereof.
The term “alkyl” as used herein typically means a linear or branched saturated hydrocarbon radical having 1-8 carbon atoms and includes, e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2,2-dimethylpropyl, n-hexyl, n-heptyl, n-octyl, and the like. Preferred are (C1-C5)alkyl groups, more preferably (C1-C3)alkyl groups, i.e., methyl, ethyl, n-propyl, and isopropyl.
The term “(C1-C5)alkylC(O)O— esters” as used herein refers to a molecule wherein the group (C1-C5)alkyl-COO— is linked, via the carboxylic group thereof, to a group such as (C1-C8)alkyl, (C2-C12)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having from 4-9 carbon atoms, 1-methyl-1-(alkanoyloxy)-ethyl having from 5-10 carbon atoms, alkoxycarbonyloxymethyl having from 3-6 carbon atoms, 1-(alkoxycarbonyl-oxy)ethyl having from 4-7 carbon atoms, 1-methyl-1-(alkoxycarbonyloxy)ethyl having from 5-8 carbon atoms, N-(alkoxycarbonyl)aminomethyl having from 3-9 carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having from 4-10 carbon atoms, 3-phthalidyl, 4-crotonolactonyl, gamma-butyrolacton-4-yl, di-N,N—(C1-C2) alkylamino(C2-C3)alkyl (such as β-dimethylaminoethyl), carbamoyl-(C1-C2) alkyl, N,N-di(C1-C2)alkylcarbamoyl-(C1-C2)alkyl and piperidino-, pyrrolidino- or morpholino(C2-C3)alkyl. According to the present invention, particular such molecules are selected from (C1-C5)alkyl-C(O)O—(C1-C5)alkyl.
The terms “(C3-C8)cyclic ether” and (C3-C7)cyclic diether, as used herein, refer to a cyclic organic compound having 4-8 carbon atoms and containing an ether group, i.e., a group of the formula R—O—R wherein R each independently represents an alkyl or aryl group, and to a cyclic organic compound having 3-7 carbon atoms and containing two ether groups as defined above. Examples of such compounds include, without being limited to, furan, furfural, THF, dihydrofuran, 2-furan methanol, 2-methyl-tetrahydrofuran, 2,5-dimethyl-tetrahydrofuran, 2-methyl furan, 2-ethyl-tetrahydrofuran, 2-ethyl furan, hydroxymethylfurfural, 3-hydroxytetrahydrofuran, tetrahydro-3-furanol, 2,5-dimethyl furan, 5-hydroxymethyl-2(5H)-furanone, dihydro-5-(hydroxymethyl)-2(3H)-furanone, tetrahydro-2-furoic acid, dihydro-5-(hydroxymethyl)-2(3H)-furanone, tetrahydrofurfuryl alcohol, 1-(2-furyl)ethanol, hydroxymethyltetrahydrofurfural, dioxanes, dioxalanes, pyrans, tetrahydropyrans, dioxins, oxepane, oxypine, and isomers thereof.
The term “glycol” as used herein refers to an organic alcohol having 2-6 carbon atoms, wherein two hydroxyl groups are attached to different carbon atoms of the molecule. Non-limiting examples of glycols include ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, and the like.
In particular such embodiments, the solvate forming solvent is selected from a formamide, acetone, TBME, THF, acetonitrile, nitromethane, pyridine, ethylene glycol, cumene, methyl acetate (MeOAc), ethyl acetate (EtOAc), isopropyl acetate, methanol (MeOH), ethanol (EtOH), IPA, n-propanol, n-butanol (n-BuOH), 1,4-dioxane, or a mixture thereof such as a mixture of IPA and cumene. More particular such embodiments are those wherein the solid crystalline form comprises about 0.1 to about 1.1, preferably about 0.5 to about 1.0, mol of formamide, acetone, TBME, THF, acetonitrile, nitromethane, pyridine, ethylene glycol, cumene, MeOAc, EtOH, IPA, or 1,4-dioxane per about 1 mol apomorphine free base.
In certain specific embodiments, the present invention provides a solid crystalline form of apomorphine free base or a hydrate or solvate thereof, having an XRPD pattern equivalent to that of
In certain embodiments, the present invention provides a solid crystalline form of apomorphine solvate as defined above, wherein the solvate forming solvent is (C1-C8)alkanol, e.g., methanol, ethanol, propanol, IPA or n-butanol, but preferably IPA. In particular such embodiments, the invention provides a solid crystalline form of apomorphine free base•IPA solvate wherein the IPA is about 15% to about 25%, about 16% to about 20%, about 17% to about 19%, or about 18% to about 19%, e.g., about 18.0%, 18.1%, 18.2%, 18.3%, 18.4%, 18.5%, 18.6%, 18.7%, 18.8%, 18.9% or 19.0%, by weight of the crystal. In other particular such embodiments, the invention provides a solid crystalline form of apomorphine free base•IPA mono-solvate, i.e., a solid crystalline form of apomorphine solvate, which comprises about 1 mol of IPA per about 1 mol apomorphine free base.
Such solid crystalline forms, e.g., an apomorphine free base/IPA crystals, may have enhanced stability against discoloration or decomposition relative to amorphous apomorphine free base. In addition, such crystals may desolvate at a temperature of about 110° C.; or melt at a temperature of about 204° C. Solid crystals of apomorphine solvate wherein the solvate forming solvent is (C1-C8)alkanol may absorb less than 0.1% w/w water from the air when allowed to equilibrate, as measured by Gravimetric Vapour Sorption (GVS), from 0% to about 90% relative humidity and 25±0.1° C.; or may contain about 0.2% w/w water or less.
In certain specific embodiments, the present invention provides a solid crystalline form of apomorphine solvate wherein the solvate forming solvent is (C1-C8)alkanol, having an XRPD pattern with peaks at:
In other specific embodiments, the present invention provides a solid crystalline form of apomorphine solvate wherein the solvate forming solvent is (C1-C8)alkanol, having an XRPD pattern equivalent to that of
In another aspect, the present invention provides a liquid formulation produced, or obtained, by dissolving a solid crystalline form of apomorphine free base or hydrate, solvate, or co-crystal thereof as disclosed herein, e.g., said solid crystalline form of apomorphine free base or solvate thereof, in a solvent. The liquid formulation of the invention may further comprise an antioxidant, i.e., an agent that inhibits the formation of oxidation products, such as an o-quinone scavenger, a tyrosinase inhibitor, a Cu+2 chelator and/or a tetrahydroquinoline.
Examples of o-quinone scavengers include, without being limited to, ascorbic acid, an ascorbate such as Na-ascorbate, ascorbic acid-6-palmitate, L-cysteine, N-acetyl cysteine (NAC), glutathione (GSH), or a mixture thereof.
Examples of tyrosinase inhibitors include, without limiting, captopril, methimazole, quercetin, arbutin, aloesin, N-acetylglucoseamine, retinoic acid, α-tocopheryl ferulate, Mg ascorbyl phosphate (MAP), substrate analogues, e.g., sodium benzoate, or L-phenylalanine, or a mixture thereof.
Examples of Cu+2 chelators include, without being limited to, Na2-EDTA or Na2-EDTA-Ca.
Other antioxidants that may be included in a liquid formulation as disclosed herein are dimercaptosuccinic acid (DMSA), diphenylamine (DPA), trientine-HCl, dimercaprol, clioquinol, sodium thiosulfate, triethylenetetramine (TETA), tetraethylene pentamine (TEPA), curcumin, neocuproine, tannin, cuprizone, sulfite salts such as sodium hydrogen sulfite or sodium metabisulfite, di-tert-butyl methyl phenols, tert-butyl-methoxyphenols, polyphenols, tocopherols, ubiquinones, or caffeic acid.
Further antioxidants that may be included in a liquid formulation as disclosed herein are thiols such as aurothioglucose, dihydrolipoic acid, propylthiouracil, thioredoxin, glutathione, cysteine, cystine, cystamine, and thiodipropionic acid; sulphoximines such as buthionine-sulphoximines, homo-cysteine-sulphoximine, buthionine-sulphones, and penta-, hexa- and heptathionine-sulphoximine; metal chelators such as α-hydroxy-fatty acids, palmitic acid, phytic acid, lactoferrin, citric acid, lactic acid, malic acid, humic acid, bile acid, bile extracts, bilirubin, biliverdin, ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), and diethylenetriaminepentaacetic acid (DTPA); sodium metabisulfite, vitamins such as vitamin E, vitamin C, ascorbyl palmitate, Mg ascorbyl phosphate, and ascorbyl acetate; phenols such as butylhydroxytoluene, butylhydroxyanisole, ubiquinol, nordihydroguaiaretic acid, and trihydroxybutyrophenone; benzoates such as coniferyl benzoate; uric acid; mannose; propyl gallate; selenium such as selenium-methionine; stilbenes such as stilbene oxide and trans-stilbene oxide; or combinations thereof.
A liquid formulation contemplated herein may thus comprise one or more antioxidants selected from ascorbic acid, an ascorbate such as Na-ascorbate, L-cysteine, NAC, GSH, Na2-EDTA, Na2-EDTA-Ca, or a combination thereof. In particular embodiments, the liquid formulation comprises ascorbic acid, ascorbic acid-6-palmitate, sodium bisulfite, or a combination of ascorbic acid and another antioxidant, e.g., a cysteine such as L-cysteine or NAC. The ratio of ascorbic acid to the other antioxidant, e.g., L-cysteine or NAC, may exist at a particular weight-to-weight ratio such as about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, or about 10:1.
In certain embodiments, the liquid formulation disclosed herein is a pharmaceutically acceptable liquid formulation also referred herein to as a pharmaceutical composition, i.e., a liquid formulation as disclosed in any one of the embodiments described above, when further comprising one or more pharmaceutically acceptable carriers or excipients.
The term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” as used herein refers to any and all solvents, dispersion media, preservatives, antioxidants, coatings, isotonic and absorption delaying agents, surfactants, and the like, that are compatible with pharmaceutical administration. Examples of such excipients include, without limiting, Tween-80, Tween-60, Tween-40, Tween-20, N-methylpyrrolidone (NMP), or polyvinylpyrrolidone (PVP), EDTA or salts thereof, cysteine, N-acetylcysteine, sodium bisulfite, and mixtures thereof. The use of such media and agents for pharmaceutically active substances is well known in the art. The pharmaceutical compositions disclosed herein may contain other active compounds providing supplemental, additional, or enhanced therapeutic functions.
The terms “pharmaceutically acceptable” and “pharmacologically acceptable” as used herein refer to molecular entities and compositions that do not produce an adverse, allergic, or other untoward reaction when administered to an animal or human, as appropriate. For human administration, preparations should meet sterility, pyrogenicity, general safety, and purity standards as required by a government drug regulatory agency, e.g., the United States Food and Drug Administration (FDA) Office of Biologics standards.
The liquid formulations and pharmaceutical compositions disclosed herein may be liquid solutions, i.e., substantially homogeneous liquid mixtures at room temperature (e.g., 25° C.). In particular embodiments, the liquid formulations and pharmaceutical compositions disclosed herein are substantially aqueous.
In certain embodiments, the liquid formulations and pharmaceutical compositions of the present invention are stable for at least 24 hours, 48 hours, or more, i.e., for 1, 2, 3, 4, 5, 6, or 7 days, 1 week, 2 weeks, 1 month, 2 months, or more, at room temperature, e.g., at any temperature in the range of 18° C. to 30° C., e.g., at 25° C.
In certain embodiments, the liquid formulations and pharmaceutical compositions of the present invention have substantially no precipitation of solids for at least 24 hours, 48 hours, or more, i.e., for 1, 2, 3, 4, 5, 6, or 7 days, 1 week, 2 weeks, 1 month, 2 months, or more, at room temperature, e.g., at any temperature in the range of 18° C. to 30° C., e.g., at 25° C.
The pharmaceutical composition of the present invention may be formulated for any suitable route of administration, e.g., for subcutaneous, transdermal, intradermal, intravenous, intraarterial, intramuscular, intraperitoneal, intrathecal, intrapleural, intratracheal, intranasal, sublingual, or buccal administration.
In yet another aspect, the present invention relates to a method of treating a neurological or movement disease or disorder, or a condition associated therewith, in a patient in need thereof, comprising administering to said patient a liquid formulation as disclosed herein. Examples of neurological or movement diseases or disorders include, without being limited to, Parkinson's disease, Alzheimer's disease, and akinesia, and non-limiting examples of conditions associated with neurological or movement diseases or disorders include alcoholism, opiate addiction, and erectile dysfunction. In a particular embodiment, the invention thus relates to a method of treating Parkinson's disease in a patient in need thereof, comprising administering to said patient a liquid formulation as disclosed herein.
In still another aspect, the present invention relates to a liquid formulation as disclosed herein, for use in the treatment of a neurological or movement disease or disorder, or a condition associated therewith. In a particular embodiment, the invention thus relates to a liquid formulation as disclosed herein, for use in the treatment of Parkinson's disease.
In a further aspect, the present invention relates to a method of producing a solid crystalline form of apomorphine free base or a solvate thereof as disclosed herein, said method comprising:
As shown herein and defined by the method of the present invention, the disclosed solid crystalline forms of apomorphine free base or solvate thereof are produced starting from apomorphine hydrochloride, which is dissolved in a solvent. Yet, it should be understood that such solid crystalline forms may further be produced, following a method similar to that disclosed herein, wherein the apomorphine hydrochloride is replaced by apomorphine hydrobromide.
In certain embodiments, the solvent dissolving the apomorphine hydrochloride and optionally said antioxidant in step (a) of the method of the invention is a formamide, acetone, TBME, THF, acetonitrile, nitromethane, pyridine, ethylene glycol, cumene, MeOAc, EtOAc, isopropyl acetate, MeOH, EtOH, IPA, n-propanol, n-BuOH, 1,4-dioxane, or a mixture thereof. In particular embodiments, said solvent is MeOH, EtOH, IPA, n-propanol, n-BuOH, or dioxane, preferably IPA.
In particular embodiments, the method of the present invention thus comprises, in step (a), dissolving apomorphine hydrochloride and, optionally, said antioxidant, in IPA, to thereby obtained, following step (c), a solid crystalline form of apomorphine*IPA solvate.
In certain embodiments, the base contacted in step (b) of the method of the invention with the solution obtained in step (a) has a pKa higher than that of apomorphine, i.e., higher than 8.9. Such a base may be selected from a (C1-C8)amino alcohol also referred to as (C1-C8)alkanolamine, i.e., an organic compound having an alkane backbone of 1-8 carbon atoms, which contain both hydroxyl and amino (—NH2, —NHR, and —NR2) functional groups. Particular such bases include, without being limited to, pyrrolidine, piperidine, 2,2,6,6-tetramethylpiperidine, diethylamine, ethanolamine, 2-(methylamino) ethanol, ethanolamine, 2-amino-1-propanol, 3-amino-1-propanol, alaninol, serinol, 2-amino-1-butanol, 4-amino-1-butanol, arginine, or N-methyl dicyclohexyl amine. In a particular embodiment, said base is 3-amino-1-propanol.
In certain embodiments, step (a) of the method of the invention comprises dissolving apomorphine hydrochloride in a solvent as defined above. In other embodiments, the solid crystalline form produced comprises at least one antioxidant, and step (a) of the method of the invention thus comprises dissolving apomorphine hydrochloride and said antioxidant in a solvent as defined above. The antioxidant may be any antioxidant as defined above, or a mixture of such antioxidants. In particular embodiments, the antioxidant is an ascorbate-based antioxidant such as ascorbic acid-6-palmitate. More particular such embodiments are those where the antioxidant is ascorbic acid-6-palmitate, and the amount of antioxidant dissolved in the solvent is about 0.001% to about 6%, about 0.001% to about 5%, about 0.001% to about 4%, about 0.001% to about 3%, about 0.001% to about 2%, about 0.001% to about 1%, about 0.005% to about 2.5%, about 0.005% to about 2.0%, about 0.005% to about 1.5%, or about 1%, 2%, 3%, 4%, 5% or 6%, by weight, relative to the amount of apomorphine hydrochloride.
In certain embodiments, at least part of the method of the present invention is performed under a flow of an inert gas. In particular such embodiments, the method of the invention is aimed at producing a solid crystalline form of apomorphine*IPA solvate, and step (a) of the method comprises dissolving apomorphine hydrochloride and, optionally, said antioxidant, in IPA, following which the solution is placed under a flow of nitrogen.
In certain embodiments, step (a) of the method of the present invention includes heating the components, i.e., said solvent, apomorphine hydrochloride, and optionally said antioxidant. In particular such embodiments, the method of the invention is aimed at producing a solid crystalline form of apomorphine•IPA solvate, and step (a) includes heating the components to a temperature of about 55° C. to about 83° C.
In certain embodiments, the solution obtained in step (b) of the method of the present invention, which contains the apomorphine free base or solvate thereof, is a homogenous solution. In other embodiments, the solution obtained in step (b) is filtered prior to step (c), i.e., before subjecting to conditions that result in crystallization of the apomorphine free base or solvate thereof.
In certain embodiments, the crystallization step (c) of the method of the present invention comprises gradual cooling over 1 to 24 hours to initiate crystallization. In particular such embodiments, step (c) comprises gradual cooling from approximately 82° C. to 68-72° C. for 1-2 h, and then to approximately 18-23° C. for 3-10 hours.
In certain embodiments, the crystallization step (c) of the method of the invention further comprises seeding the solution with seed crystals to initiate crystallization of the solution.
It should be understood that a variety of crystallization methods and techniques may be used in accordance with the disclosed method. For example, the crystallization of the apomorphine free base or solvate thereof in step (c) may be performed by diffusion techniques; evaporative crystallization, e.g., at a high partial pressure of an inert gas such as nitrogen; slow evaporation under reduced pressure; or a classical crystallization. The crystallization may also be performed by rapid, ballistic, or shock cooling, e.g., to 0° C. or −78 C°.
In yet a further aspect, the present invention relates to a crystalline form of apomorphine free base or solvate thereof produced, or obtained, by a method as disclosed herein.
In still a further aspect, the present invention provides a kit comprising (i) crystals or co-crystals of apomorphine free base or a hydrate or solvate thereof, or a formulation obtained by dissolving said crystals or co-crystals, and optionally an additional therapeutic agent such as levodopa, carbidopa, or entacapone, in a solvent; and optionally (ii) instructions for use. Such a formulation may be liquid as disclosed herein, or in the form of a lyophilized powder that can be reconstituted into a liquid formulation. The formulation may be designed for administration by any suitable route such as, but not limited to, subcutaneously, transdermally, intradermally, intravenously, intraarterially, intramuscularly, intraperitoneally, intrathecally, intrapleurally, intratracheally, intranasally, sublingually, or buccally; or may be designed for transdermal administration and form a part of a transdermal patch.
In certain embodiments, a kit as disclosed herein comprises one, two, three, or more pre-filled cartridges, each containing a liquid formulation as disclosed herein, suitable for use by a patient or a physician. Each such pre-filled cartridge may contain a disclosed liquid formulation comprising a single dose, i.e., a dose suitable for a single administration to a patient, of an apomorphine free base or a solvate thereof, and optionally an additional therapeutic agent, e.g., levodopa, carbidopa, or entacapone.
The invention will now be illustrated by the following non-limiting Examples.
An aqueous procedure was used to isolate apomorphine free base using the following steps. Apomorphine hydrochloride (3.1 g, 1.0 wt) was dissolved in 0.1% w/w aqueous sodium metabisulfite solution (200 ml, 67 vol). 1N aqueous sodium bicarbonate solution was added (33 ml, 11 vol) at approximately 25° C. Stirring continued for approximately 30 minutes at approximately 25° C. The resulting precipitate was filtered using Whatman paper #43 ø 110 mm (1 L Buchner flask) using a vacuum pump. The precipitate was washed with water (2×100 ml, 2×33 vol) and dried under a flow of N2 using a vacuum pump for 4 h. The expected yield was 83.4%. An improved result was obtained by cooling the precipitate for an additional 20 minutes at approximately 4° C.
A crystallization study of the free base was conducted (A0513-36) focusing on removal of color, filterability, and determination of whether the solid was crystalline or amorphous. The apomorphine free base used was an amorphous residue isolated from the resin procedure (A0513-32) described below. This solid was highly colored, underscoring the need for a procedure capable of removing any colored impurities in the solid. Solvents and mixtures of solvents were investigated for crystallization of the free base. The results are summarized in Table 1 below.
Crystallization solvent candidates that resulted in solids include IPA, methyl acetate (MeOAc), ethyl formate, ethanol (at low volumes), ethyl acetate (EtOAc), and isopropyl acetate (i-PrOAc). Crystallization solvent candidates that result in a gum or highly colored solids include: toluene, chlorobenzene, n-butylacetate (n-BuOAc), isobutyl acetate (i-BuOAc), methyl isobutylketone (MIBK), and anisole. Solvents include methanol, n-butanol, THF, MEK. Anti-solvents include TBME, heptane, diisopropyl ether (DIPE), and cumene.
The most promising solids isolated from crystallization solvent candidates were investigated by microscope under plain- and cross-polarized light. These were isolated in good yield and were white to off-white solids (
Four samples (B, G, H and O) were transferred to a vacuum oven and dried at 45° C. for 24 h. Post-drying, solvent contents were 23.4% w/w dioxane, 18.6% w/w IPA, 20.0% w/w MeOAc and 7.8% w/w EtOH, respectively. Crystals isolated from dioxane and IPA were approximately mono-solvates and in the case of EtOH a hemi-solvate. Sample G (from IPA) was heated to 75° C. for 17 h under vacuum, at which point IPA remained at 17.4% w/w, indicating that the solvent is bound in the crystal. MeOAc and IPA resulted in the whitest material, and they had a similar level of crystallinity. These solvents were entrained in the crystal at approximately 1 molar equivalents (eq.), confirming that they were stoichiometric solvates.
Crystals isolated from IPA and 1,4-dioxane were investigated by DSC to determine the temperature at which solvent would be released. IPA is released at >110° C., and dioxane at >120° C. IPA and MeOAc allow full dissolution of the free base at moderate temperature and poor solubility on cooling to ambient temperature. Controlling the rate of cooling can therefore allow control over the rate of crystal growth and thus control of the filtration, which is problematic when following the aqueous route.
Significantly, both IPA and MeOAc allow removal of the green color from the product. IPA was selected as the lead solvent candidate for the final crystallization.
Samples of free base isolated from water, IPA, and MeOAc were retained open to air and the color visually monitored (
A portion of apomorphine free base was synthesized utilizing the supplied aqueous method (A0513-02-01). This material was used in 50 mg portions (A0513-06).
The free base is fully soluble in 5 vol of either MeOH, EtOH, or n-BuOH. Of the solvents screened, n-BuOH appeared promising as a candidate for a reaction solvent, as it did not dissolve the HCl salt (in up to 20 vol at reflux) and could potentially offer both a phase separation with water and an azeotrope with water to dry the organic phase (if required). Methanol was the only solvent that dissolved any significant amount of the HCl salt. The free base precipitated as an oil from most solvents, particularly after heating. An apparently crystalline solid formed following dissolution of the free base and cooling in 1,4-dioxane.
Salt release was accomplished using: 1) basic resins where the resin would be filtered from a solution of the product; 2) organic bases to remove the base hydrochloride salt by precipitation, retaining the apomorphine in solution; and 3) lipophilic bases keeping the base hydrochloride salt in organic solution and obtaining the apomorphine free base by precipitation or extraction.
To use resins, solubility of the free base in solution was required, and therefore the alcoholic solvents identified above that are able to dissolve the free base were used. To determine the required loading of resin and the most suitable solvent, a small screen was performed utilizing Amberlite IRA-402(OH) and Ambersep900 hydroxide form resins with MeOH, EtOH, and n-BuOH. To each portion of apomorphine hydrochloride (200 mg, 1.0 wt), Amberlite or Ambersep (100 mg, 0.5 wt) was added in MeOH, EtOH or n-BuOH (7.5 vol).
The noted weights of resins included the weight of water contained in the resin. Prior to use, each resin was washed with solvent (3×10 vol) to remove water. The progress of the reactions could be followed visually as the apomorphine hydrochloride was present as a white slurry (except in MeOH, in which the HCl salt has moderate solubility), while the resin was present as red beads. The white slurry disappeared as the reaction progressed.
Salt release with basic resins was possible and performed well with Amberlite IRA-402(OH) (2.5 wt, wet weight). Ambersep resin required higher loadings to effect the salt release and was therefore not investigated further. Methanol was preferred, as the HCl salt had more solubility in MeOH than the other alcohols, resulting in the fastest reaction rate.
The Amberlite IRA-402(OH) conditions in MeOH were scaled up to 10 g (A0513-32; this material was later used in the initial screening of isolation conditions, see below).
Amberlite IRA-402(OH) (25 g, 2.5 wt) was washed with MeOH (degassed, 4×100 ml, 4×10 vol); 4 washes resulted in 2.0% w/w water. Apomorphine hydrochloride (10.0 g, 1.0 wt) was charged to the resin with MeOH (degassed, 75 ml, 7.5 vol). The mixture was slurried, protected from light and air at 18-23° C. for 30 min, and filtered and washed with MeOH (2×7.5 vol). The blue/violet solution was concentrated to a residue at 45° C. on a rotary evaporator to yield 5.9 g (67%) (A0513-32-01) as a turquoise solid (yield corrected for 7.7% w/w MeOH by 1H NMR). Liquid chromatography showed 99.68% area vs. 99.73% area when released from via the supplied aqueous route. In an attempt to recover material from the resin, washes with 2 M HCl in MeOH (40 vol) were performed to yield 600 mg of a black tar.
Although a portion of apomorphine free base was generated for use in solubility screening, the recovery from the resin was poor. The yield was low and the material isolated was highly colored, which was challenging to remove by crystallization.
A base with a pKa higher than that of apomorphine (pKa 8.9) was used to release salt from apomorphine and to form its own salt (Scheme 1). Some base salts would be insoluble in a solvent which apomorphine free base would be soluble, allowing removal of the base hydrochloride by filtration.
n-BuOH (7.5 vol) was first used as a solvent because it solubilizes the free base but not the apomorphine hydrochloride. The aim was to determine which base hydrochloride salts could be removed by filtration, thus leaving behind a solution of apomorphine free base. A selection of amine bases having pKa values ranging from 10.8 to 13.2 were screened (A0513-10).
The selected base (1.1 eq) was added to a slurry of apomorphine hydrochloride (100 mg) in n-BuOH (7.5 vol). Where a solid formed, it was filtered off and the product-containing liquor analyzed and compared with the expected NMR spectrum of the free base. Where appropriate, the base hydrochloride collected by filtration was also analyzed to determine if the apomorphine hydrochloride was fully released. The results are summarized in Table 2.
The appearance of the salt release reactions is shown in
Of diisopropylamine, L-arginine, and lysine, L-arginine and lysine formed salts that were not soluble in d6-DMSO. On investigating the solid and the liquor (by NMR), the presence of the L-arginine and lysine could not be detected. Signals were detected for diisopropylamine in the NMR of both the liquors and the isolated solid, indicating that an alternate solvent to n-BuOH may be required in which the diisopropylamine hydrochloride salt is less soluble. L-Arginine was further investigated for the salt release (A0513-50) using MeOH, EtOH, and n-BuOH as reaction solvents (Scheme 2).
MeOH, EtOH and n-BuOH were added to vials (A, B, D and E) with apomorphine as shown in Table 3. HCl (11-00645, Johnson Matthey, 98.9% w/w by NMR assay vs. TCNB) (3×200 mg) and L-arginine (126 mg, 1.1 eq) were added. The reaction was stirred at ambient temperature. Vials D and E were heated to 40° C. to aid solubility of the apomorphine free base. Strong apomorphine signals were detected in the L-arginine removed by filtration by 1H NMR analysis. The solids were filtered after 2 h of stirring (n-BuOH after 16 h at 40° C.).
The amount of arginine (HCl or free base) retained in the solution of product was found to be least when ethanol was employed as the solvent. Salt release in ethanol at 40° C. was then scaled up to determine the yield more accurately and analyze the viability of the route (A0513-60).
apomorphine hydrochloride (10.0 g) and L-arginine (6.02 g, 6.0 wt, 1.05 eq) were added to a flask, EtOH (7.5 vol) was added, and the solids were slurried at ambient temperature then heated to 38-42° C. There remained a white slurry throughout. After 1 h at 40° C., 1H NMR analysis indicated that complete salt release had occurred. Therefore the slurry was filtered at 40° C. and washed with EtOH (2×5 vol). The yield was 4.28 g (48.7%) contained in ethanolic solution A0513-60-02. Assay of the solids indicated 4.53 g (51.5%) contained apomorphine.
The white solids were returned to flask under nitrogen and slurried in EtOH (7.5 vol) at 40° C. After 4 h at 40 to 45° C. the slurry was filtered and a further 1.1 g (12%) was isolated (by solution assay of the liquors). The solids were again returned to the flask and slurried in EtOH (5.0 vol) at 40° C., and after 16 h at 40-45° C. the slurry was filtered and a further 1.1 g (12%) was isolated (by solution assay of the liquors).
The reaction did not fully complete the salt release during the stir period at 40° C., or the product as the HCl salt was encapsulated by the arginine hydrochloride salt. Thus, another approach was employed.
Amines were used to release the apomorphine hydrochloride in IPA, with the objective of leaving the HCl salt of the amine used to release the apomorphine hydrochloride salt in solution with precipitation of the apomorphine free base (A0513-42). The amines chosen below released the apomorphine salt in n-BuOH but remained in solution as their HCl salts (findings from A0513-10). Samples of apomorphine hydrochloride (200 mg) were slurried in IPA (2 ml, 10 vol) and amine base (1.1 eq) added.
NMR analysis indicated that each of the samples had fully released the apomorphine salt. The samples were heated to aid mobility (see Table 4 for observations). All samples were then cooled to 18-23° C. for 2 h and aged for 1 h before filtration. The solids were washed with IPA (2×5 vol) at 18-23° C. and dried for 5 min on the filter.
After storage for 64 h, reactions B and E showed the least green coloration, and C was dark green. Each of the bases effected full salt release of the apomorphine, which precipitated in each case to yield a filterable solid. At ambient temperatures gels formed in some cases but heating resulted in mobile slurries. Therefore, adding the amine to a warm (approximately 40° C.) slurry of the apomorphine hydrochloride was expected to aid mobility and the crystallization of the product. Only reaction C with TMP resulted in significant entrainment of the base used to release the apomorphine in the product (Table 5).
From the selection of bases reported above, piperidine and 3-amino-1-propanol resulted in white apomorphine with no base detected in the isolated product by 1H NMR analysis. Because 3-amino-1-propanol is of lower toxicity, it was chosen for the scale-up reaction. Scheme 3 shows the initial salt release scale-up reaction conditions.
The 3-amino-1-propanol mediated salt release of apomorphine was scaled to 10 g in IPA (10 vol), and in order to aid the mobility of the reaction, the addition of the 3-amino-1-propanol was performed at 40-45° C. On rapid addition a full solution formed from which precipitated the product after approximately 15 min. Analysis of the liquor indicated that 80% crystallization had occurred (later reactions indicated that when the addition time of the 3-amino-1-propanol was increased, the HCl salt converted to the free base without formation of a solution). The slurry was cooled and aged over 16 h at 18 to 23° C. The solid was filtered and washed with IPA to yield a white solid in 92.0% yield corrected for 84.7% w/w assay (the combined mother liquors and washes contained 6.2% yield by assay) (17.8% w/w IPA, approximately 1 eq) 100% area by HPLC KF=0.1% w/w.
The scale-up reaction indicated that apomorphine free base mono-IPA solvate product could be isolated as a white crystalline solid. The solid was found to contain no residual 3-amino-1-propanol by 1H NMR.
The stability of the product to air oxidation is expected to increase by increasing the particle size. This reduces the total surface area of the product relative to mass and results in less crystal/air contact. A recrystallization would also be useful as a purification method if ascorbic acid-6-palmitate or 3-amino-1-propanol were entrained in the isolated product. The recrystallization of isolated apomorphine free base was performed to determine the volume of IPA required and the potential seed point if required (A0513-90-01).
On a 1.0 g scale (The material used was A0513-68-01, previously isolated from IPA in the presence of ascorbic acid-6-palmitate), the apomorphine free base was slurried in IPA (10 ml, 10 vol). The slurry was vacuum/nitrogen purged three times at ambient temperature and heated to 50° C. At this temperature, a slurry remained, and only on achieving reflux and adding further IPA (1 vol) was full dissolution obtained. On cooling, solids formed at 71° C.; based on this a seed temperature of 77-78° C. was calculated (later experiments indicated that the optimal temperature for seeding the reaction mixture was 71° C., the difference is likely to be due to the presence of the 3-amino-1-propanol hydrochloride salt). This should allow seeds to be added and not dissolve which will initiate crystal growth and potentially generate larger crystals. The slurry was warmed to 74° C. and held for 1 h before cooling to 55° C. The slurry was held for 40 min before cooling to 18-23° C. for 2 h. The slurry was filtered and resulted in a yield of 1.00 g, 83.5%, 83.5% w/w assay, 16.8% w/w IPA (some mechanical losses were made in transfer to the filter). Chemical purity (CP)=100% area.
The product was stable under the crystallization conditions over the 4 h time period, and no color was observed in the liquor. The isolated material was investigated by microscopy (
The apomorphine free base isolated as an IPA solvate was found to be relatively stable in air when compared to the aqueous route. However, the free base in solution is susceptible to aerial oxidation. To improve stability during processing, ascorbic acid-6-palmitate or BHT were assessed re: stabilizing the apomorphine free base in solution (A0513-54).
200 mg portions of apomorphine free base (isolated by the supplied aqueous route, A0513-02-01) and antioxidant (10 wt %, 20 mg) were added to vials, followed by MeOH (10 vol). The vial was flushed with air, sealed, and stirred. The reactions were compared with a control reaction having no antioxidant.
Samples were analyzed by HPLC.
After 72 h, all samples had dark-colored precipitate present. Sample B exhibited the highest purity, while samples A and C continued to degrade to early running impurities totaling >3.1% area and >4.3% area. Further impurities formed at RRT 1.09, 1.25 and to a lesser extent at RRT 1.37, 1.71, and 1.76.
The ascorbic acid-6-palmitate had a stabilizing effect on the apomorphine free base, and prevented discoloration for a limited time, likely due to full consumption of the ascorbic acid-6-palmitate.
The effect of adding ascorbic acid-6-palmitate on stability of the reaction mixture and the isolated solid was determined in a larger scale reaction (A0513-68). On a 10 g scale, ascorbic acid-6-palmitate (0.01 wt) was added to a flask with the apomorphine hydrochloride salt and IPA (10 vol). 3-amino-1-propanol (2.5 ml, 1.1 eq, 0.25 vol) was added at 40-45° C., and a slurry was present throughout. The slurry was stirred at 40-45° C. for 2 h; at this point 10% of the reaction mixture was removed for an extended stability study at 40-45° C. The remainder of the reaction was cooled and aged over 16 h to 18-23° C. The solid was filtered and washed with IPA (2×3 vol) and resulted a white solid (liquor was pale yellow). The solid was dried under a flow of nitrogen for 30 min. The yield was 7.03 g (79.8%, corrected for 18.3% w/w IPA, approximately 1 eq), 100% area by HPLC (Note: 10% of the reaction mixture was removed for the stability study, therefore this yield is at the expected approximately 90% level, and the combined mother liquor and washes contained 7.6% of theoretical yield by assay). In this material the ascorbic acid-6-palmitate and 3-amino-1-propanol in the product were below detectable levels by 1H NMR analysis.
Stability of the reaction mixture was compared with and without ascorbic acid-6-palmitate. Each reaction was held at 40-45° C. and both the color and the HPLC profile monitored. HPLC purity profiles are shown on Table 6. Photographs of the reactions after extended time periods are shown in
Effective purging of the reaction to remove air is important to obtain a stable reaction mixture at 40-45° C. Even with good purging, minor discoloration is observed over extended stir periods at 40-45° C. Ascorbic acid-6-palmitate resulted in less discoloration but little difference in HPLC profile over the time period examined.
Following the observation that a controlled cool down from reflux resulted in a larger particle size, an investigation of the stability under reflux conditions in IPA (10 vol) was performed. In this case, the base was added under reflux and the procedure compared with and without ascorbic acid-6-palmitate as an antioxidant (Table 7).
The reaction without ascorbic acid-6-palmitate was found to be unstable and resulted in a green solution after 2 h. This instability was unexpected as the recrystallization experiment discussed above, which employed high-purity apomorphine free base previously isolated from IPA, did not indicate that the product was particularly unstable. It was therefore concluded that the presence of the 3-amino-1-propanol led to the reduced stability under reaction conditions. Addition of ascorbic acid-6-palmitate to the reaction mixture from the start resulted in a clear pale yellow solution after 2 h at reflux, and the solid later isolated was whiter than that isolated in the absence of the ascorbic acid-6-palmitate. The use of 0.01 wt % of the antioxidant was therefore added to the process. This addition was shown to have no detrimental effect on the reaction and to be removed beyond the limit of 1H NMR detection levels on isolation from IPA. Although the color difference was significant, the difference in the HPLC profile was minor; the isolated yields were also comparable at approximately 85% each.
Samples of solids isolated from an IPA recrystallization of the free base, salt released via the 3-amino-1-propanol route were monitored by their color change and their HPLC profile. The HPLC profiles are described in Table 8. Photographs of the color changes are shown in
The isolated product IPA solvate was found to be stable at ambient conditions when exposed to air and light when isolated from reactions with 3-amino-1-propanol used as the base. This is in contrast to materials isolated in this manner using alternative bases (i.e., pyrrolidine, diethylamine, N-methyl dicyclohexylamine, or TMP). This suggests that the base used, and efficient removal of it, and/or the highly crystalline form of the solvate, can be important to the stability of the IPA solvate.
An optimized non-aqueous procedure is provided below and shown in Scheme 4.
The procedure above was used to prepare a 30 g sample for use in accelerated stability testing. The preparation was performed as described above (A0513-126).
Key points of note: (i) No exotherm was noted on addition of the 3-amino-1-propanol. (ii) During the crystallization phase of the preparation the slurry was heated to 82° C. and full dissolution was obtained. During the cooling step to 71° C. for 1 h the precipitate was noted at 72° C. (iii) The slurry was cooled to 18-23° C. at the rate of approximately 10° C./h. After 5 h the temperature was 21° C. and the slurry was aged for 2 h. (iv) Output: 34.11 g, 86.1% corrected for 17.9% w/w IPA (A0513-126-01). (v) The process performed as expected except for a marginally suppressed yield, which may be due to a shorter age period at ambient temperature. The material was isolated as a highly crystalline solid (approximately 100×250 μm in size; see
The aqueous route described above was performed to prepare a 30 g sample. Scheme 5 shows the reaction conditions for the aqueous route. The preparation was performed as described above (A0513-132; XRPD data shown in
Key points of note: (i) The precipitate was washed with degassed water (2×1.5 L, 2×33 vol) and dried under N2. Drying of the filter cake was slow considering the scale: 4 h KF=66% w/w water; 6 h KF=23% w/w water; 8 h KF=22% w/w water; 10 h KF=4% w/w water; 12 h KF=2.3% w/w water. (ii) Drying was stopped. Following removal of the solid from the filter, the solid was homogenized and KF was found to be 5.7% w/w. The solid was returned to the filter for further drying: 1 h KF=4.0% w/w; 2 h KF=3.8% w/w; 2.5 h KF=4.3% w/w drying was stopped as a minor amount of pale green was noted in the filter cake; Output: 29.7 g, 75% uncorrected for 4.3% w/w water (A0513-136-07).
The isolated material was crystallized from IPA.
A0486-178—Recrystallization of Crude A0513-136-07 from IPA
The aqueous route performed as expected up to initial isolation. The filtrations were acceptable; however, the filter cake was sticky, difficult to paste down, and cracked heavily. The filter cake was difficult to dry and dried non-uniformly.
During the IPA crystallization of the isolated crude a quantity of insoluble solids was encountered during the crystallization. The insoluble substance has an apomorphine-related HPLC and 1H NMR profiles, and LCMS indicates the mass ion for apomorphine. A clarification in 30 vol of IPA was included in the process to remove the solids and mitigate any affect the solids may have on the crystallization and purity of the product. The crystallization resulted in lower recovery (71% vs. expected 85-90%, the remaining mass was found in the liquors). The crystalline form by XRPD (
Following a screen of organic bases and basic resins, a scalable non-aqueous salt release for apomorphine hydrochloride was developed using 3-amino-1-propanol as the organic base. The 3-amino-1-propanol hydrochloride is removed to the mother liquor on isolation of the apomorphine free base from IPA. The addition of antioxidants was investigated and it was concluded that the addition of 0.1 wt % of ascorbic acid-6-palmitate resulted in a reaction mixture with a greater stability. The product isolated via the non-aqueous route is a highly crystalline mono-IPA solvate; the isolation was developed to control the crystallization which gave control over residual 3-amino-1-propanol and ascorbic acid-6-palmitate and generated a large particle size.
In comparison with the aqueous method, the developed route results in a product which is isolated in higher overall yield, 86% vs. 75%. By contrast, if the current IPA crystallization is applied to the material derived from the aqueous route, the crystallization results in an overall process yield of 50%.
Apomorphine free base batches: A0513-002-03, A0513-132-07, and A0530-020-01 were used for the salt investigation. Apomorphine•1*IPA batches: A0513-126-01, A0530-020-01, and A0526-010-A1 were used for the polymorph investigation.
Apomorphine free base was investigated with solvents that belonged to European Pharmacopoeia/ICH Classes 2 and 3. Twenty six crystalline or partially crystalline solids were identified; the remaining isolated solids were amorphous by XRPD. The diffraction patterns of the 26 crystalline solids were then compared and 13 of these were found to be consistent with a solid phase impurity described below that was recovered during the crystallization of the 30 g batch prepared via the aqueous route. The 12 remaining crystalline and partially crystalline solids (not including the product isolated from isopropanol, A0530-010-T1), that were not concordant with either apomorphine•1*IPA Form A or the impurity (A0486-178-A1), are believed to be novel. Characterization data of the solvents diffraction patterns and assumed temperatures of solvent release are shown in
Solvates from the Polymorph Investigation
During suspension equilibrations of apomorphine•1*IPA in a variety of solvents, under aqueous and anhydrous conditions (described below), five crystalline solvates were unintentionally generated via exchange of the isopropanol:
The main diffraction peaks of the ethanol and TBME solvates corresponded to those first identified in the solvate screen. Whilst the main diffraction peaks of the acetone and THF solvates were markedly different from those identified in the solvate screen these differences were attributed to possible polymorphism.
An impurity was first isolated during the 30 g crystallization of apomorphine•1*IPA from isopropanol, prepared via the aqueous route. The impurity was isolated from 13 of the 26 crystalline solids generated during the solvate screen. The identification of this impurity was attempted.
Analytical data obtained from the impurity (A0486-178-A1) was compared to the corresponding analysis performed on the recrystallized demonstration batch of apomorphine•1*IPA (A0526-010-A1). The findings from this investigation are summarized below:
To establish if treatment of apomorphine•1*IPA with carbon dioxide under aqueous conditions resulted in the formation of an ionic carbon dioxide adduct (apomorphine hydrogen carbonate or disproportionated into apomorphine carbonate) or generated a formal covalent cyclic carbonate under anhydrous conditions (Scheme 6), the following experiments were performed:
A solution of apomorphine•1*IPA in isopropanol (50.0 vol) at 50-55° C. was quenched by a single pellet of dry ice, the product was isolated by filtration (65% theoretical) and was consistent with apomorphine•1*IPA, Form A by XRPD. The structure was concordant with apomorphine by 1H NMR and contained one equivalent of isopropanol.
A suspension of apomorphine•1*IPA, Form A, was stirred in IPA/water (10.0/9.0, v/v, 11.0 vol) at 18-23° C., under CO2 at balloon pressure for 24 h, the product was consistent with apomorphine•1*IPA by XRPD. The structure was concordant with apomorphine by 1H NMR and contained one equivalent of isopropanol.
Two principal forms of apomorphine were incorporated into the polymorph investigation; the form generated by crystallization from isopropanol and designated as Form A (Table 9) and the form derived by precipitation from water (consistent with amorphous material). Apomorphine•1*IPA with not less than 16.5% w/w and not more than 20.2% w/w isopropanol content by GC-HS, MET/CR/2497 was preferred.
1H NMR: conformed to molecular structure
1H NMR assaya (acquired against TCNB as
The salt release of apomorphine hydrochloride in the presence of antioxidant ascorbic acid-6-palmitate is performed in IPA by treatment with 3-amino-1-propanol. Apomorphine•1*IPA then crystallizes from solution as prisms and 3-amino-1-propanol hydrochloride remains in solution. A second crystallization of apomorphine•1*IPA from IPA is then performed to upgrade the appearance of the material.
The salt release of apomorphine hydrochloride in the presence of antioxidant sodium metabisulphite is performed under aqueous conditions by treatment with sodium hydrogen carbonate. Amorphous apomorphine then precipitates from solution.
The polymorph investigation consisted of techniques to induce stable forms such as suspension equilibrations and crystallizations in conjunction with techniques intended to promote kinetic forms e.g., ballistic cooling and co-solvent precipitations, etc.
The DSC of apomorphine•1*IPA (Form A) contained a large endothermic event with onset 110° C. (
To confirm that apomorphine•1*IPA (Form A) and the related desolvated form (apomorphine•0*IPA) were not isostructural by XRPD, a specimen of Form A IPA solvate (
The freshly dehydrated specimen was then expressed from the crucible under nitrogen and analyzed by 1H NMR and XRPD (
The dehydrated specimen that had undergone thermal release of IPA (A0526-010-A1 140° C.) was not consistent with authentic Form A IPA solvate (A0526-010-A1), indicating that desolvation had altered the crystal structure. Absence of the IPA was confirmed by 1H NMR analysis.
To gain better understanding of the small exothermic event present on the DSC (approximately 140-160° C.,
The endotherm observed by DSC with onset 110° C. corresponded to release of a liquid from the crystal by thermal microscopy, attributed to IPA desolvation of apomorphine•1*IPA (
The thermal microscopy analysis was repeated under a fast stream of nitrogen. The cooling effect of the nitrogen affected the temperature readings. Release of IPA was observed at approximately 134° C. A transition occurred >200° C. and needles began to grow. The needles proliferated until they melted and had cleared by 260° C. The chemical identity of the specimen at this temperature was not known. When the melt was exposed to the air, rapid discoloration occurred.
Specimens of amorphous apomorphine (5 mg, A0530-020-01) and apomorphine•1*IPA (Form A), (A0526-010-A1) were compressed under a pressure of 10 tones (10 t, 10×1000 kilograms), under nitrogen at 18-23° C. and maintained under these conditions for 1 h. After treatment all of the specimens became ‘glassy’ in appearance. XRPD analysis of the recovered specimens showed an increased amorphous content (A0526-010-A1) and a corresponding loss of resolution of the main reflections (Table 10).
Suspension equilibration under anhydrous or aqueous conditions is a thermodynamic technique. The technique was applied to apomorphine•1*IPA (Form A) to determine if the phase evolved into a more stable modification.
Sixteen portions of apomorphine•1*IPA (approximately 100 mg, 1.0 wt.), antioxidant (0.01 wt.) and the appropriate solvents (1000-2000 μl, 10.0-20 vol) were charged to 16 separate vessels and stirred for 5-10 days at 40-45° C. If the majority of the solid dissolved, the solution was concentrated by approximately one half under nitrogen and if a suspension was generated then equilibration was resumed. If no such suspension was generated then the experiment was repeated using an equal volume of an inert diluent such as n-heptane, if the majority of the solids still dissolved under these conditions then the experiment was abandoned.
After this time the products were isolated by filtration, washed with recycled maturation solvent, dried under a stream of nitrogen at 18-23° C. and analyzed by XRPD for evidence of alternative crystalline forms and their compositions determined by 1H NMR.
No alternative phases to the apomorphine•1*IPA prepared in the demonstration batch were identified. Typically, IPA was removed or partially exchanged from apomorphine•1*IPA by the suspension equilibration solvent. Suspension equilibrations of apomorphine•1*IPA in IPA under anhydrous (A0505-080-N1) and aqueous (A0505-090-N1) conditions generated materials that were consistent with the authentic starting material by XRPD.
Suspension equilibrations in anhydrous n-heptane (A0505-080-J1) generated a mixture of unchanged apomorphine•1*IPA solvate and a new phase (believed to be crystalline apomorphine free base). Treatment of apomorphine•1*IPA with cumene (isopropylbenzene) generated mixed solvates (A0505-080-E1 and A0505-090-E1) that were isostructural with the input material; the solvent stoichiometries were disrupted by the presence of amorphous free base. Five crystalline solvates were obtained by exchange of the solvated isopropanol and were characterized by 1H NMR, XRPD and DSC:
The effect that different rates of cooling had on solutions of apomorphine in IPA were examined to show that crystallization promoted by rapid cooling does not alter the physical phase of the isolated apomorphine•1*IPA product. The experiments are described below and the results are summarized in Table 11.
Apomorphine free base (150 mg, 1.0 wt), ascorbic acid-6-palmitate (1.5 mg, 0.01 wt) and IPA (1650 μl, 11.0 vol) were stirred at 18-23° C. The suspension was sonicated to degas and placed under a flow of nitrogen at 18-23° C. The suspension was heated to approximately 80-85° C., to effect dissolution. The solution was stirred for 15-20 minutes, cooled until the cloud point was observed approximately 70° C. and aged at the cloud point temperature over 16-20 h. The suspension was filtered, washed with IPA (2×450 μl, 2×3 vol) and dried under a flow of nitrogen at 18-23° C. for at least 30 min.
The experiment was repeated as above (A0505-124-A1) except the hot solution at 80-85° C. was plunged into ice water at 0° C. The suspension was aged at this temperature for 1-2 h, filtered, washed with isopropanol (2×450 μl, 2×3 vol) and dried under a flow of nitrogen at 18-23° C. for at least 30 min.
The experiment was repeated according to A0505-124-A1, except the hot solution at 80-85° C. was plunged into carbon/dioxide acetone at −78° C. The suspension was filtered immediately, washed with IPA (2×450 μl, 2×3 vol) and dried under a flow of nitrogen at 18-23° C. for at least 30 min.
The isolated products were consistent with apomorphine•1*IPA (Form A) and the yields ranged from 76-81% (theoretical).
The crystallization procedure was able to tolerate natural and accelerated rates of cooling and still generate apomorphine•1*IPA as the isolated product (Table 11). The isolated samples exhibited good crystalline diffraction patterns (e.g.,
The effect of seeding solutions of apomorphine in IPA was examined to show that Form A is still isolated even after nucleation by alternative physical forms of apomorphine.
Apomorphine free base (150 mg, 1.0 wt), ascorbic acid-6-palmitate (1.5 mg, 0.01 wt) and isopropanol (1650 μl, 11.0 vol) were stirred at 18-23° C. The suspension was sonicated to degas and placed under a flow of nitrogen at 18-23° C. The suspension was heated to approximately 80-85° C. to effect dissolution. The solution was stirred for 15-20 minutes, cooled until the cloud point was observed approximately 70° C. The temperature was increased by approximately 5° C. to re-dissolve the solids and apomorphine seeds (3.0 mg, 2% w/w) were charged to the solution. The fates of the seeds were observed. The suspension was aged at this temperature for 1-2 h and cooled to 18-23° C. The suspension was filtered, washed with IPA (2×450 μl, 2×3 vol) and dried under a flow of nitrogen at 18-23° C. for at least 30 min.
Seeds Used:
Apomorphine free base (A0530-020-01, generated via the aqueous method), apomorphine•1*IPA (A0505-096-A1, generated via evaporation from IPA and exhibited erroneous reflections by XRPD at 2Theta 24.2° and 20.7°) and apomorphine hydrochloride.
The isolated products were consistent with apomorphine•1*IPA (Form A) and the yields ranged from 75-82% (theoretical).
The crystallization procedure was able to tolerate seeding at 2% w/w by different forms and phases of apomorphine and still generate apomorphine•1*IPA (Table 12).
These experiments were performed to determine if metastable kinetic forms of apomorphine•1*IPA were generated by evaporation of the corresponding solution in IPA.
Apomorphine free base (150 mg, 1.0 wt), ascorbic acid-6-palmitate (1.5 mg, 0.01 wt) and IPA (150000, 100.0 vol) were charged to a capped vessel and stirred at 18-23° C. under nitrogen. The slurry was heated to approximately 80-85° C. until full dissolution had taken place. The solution was cooled to 40° C. and left to stand under a very slow stream of nitrogen whilst evaporated to dryness over 18 h to give a pale brown solid.
The pale brown solid was recovered in quantitative yield and contained isopropanol (17.3% w/w) that corresponded to 1.0 to 0.91 apomorphine:IPA stoichiometry (by 1H NMR). Some differences were evident by XRPD at 2Theta 24.2° and 20.7° (
The product obtained from experiment A0505-096-A1 (100 mg) was stirred at 40 to 45° C. for 3 days under nitrogen, filtered, washed with recycled liquors to give 78 mg (78% theoretical) as an off-white solid. XRPD analysis (
As above, except the solution was left to stand under a very slow stream of nitrogen whilst cooling to 18-23° C. until evaporated to dryness. Pale brown solid was recovered in quantitative yield. The diffraction pattern (
The purpose of these experiments were to determine if metastable kinetic forms of apomorphine•1*IPA were generated by precipitation promoted by co-solvent addition to an IPA solution.
Sixteen portions of apomorphine free base (150 mg, 1.0 wt.) and ascorbic acid 6-palmitate (1.5 mg, 0.01 wt) were charged to 16 separate vessels. IPA (15000, 10.0 vol) was charged to each vessel, the vessels were capped and heated to 80-85° C. to affect full dissolution. The relevant solvent (15000, 10.0 vol) was then charged to each of the hot solutions. The mixtures were left to cool to 18-23° C., with stirring over 16-20 h and the products were isolated by filtration under nitrogen (Table 13).
No alternative phases to the apomorphine•1*IPA (Form A), prepared in the demonstration batch were identified. However, two product subsets were identified. The first set consisted of mixed occupancy solvates, isostructural with apomorphine•1*IPA (Form A) and the second set consisted of low yields of the impurity, consistent with the clarification residue (A0486-178-A1) recovered from the 30 g batch that used free base apomorphine derived from the aqueous route (See Example 10). An explanation for the presence of this impurity was that the starting material (A0530-020-01) used for the investigation was apomorphine free base that was derived from the aqueous route.
Mixed occupancy solvates were generated when IPA was partially exchanged from apomorphine•1*IPA by the precipitant solvent, except in the cases of diethyl ether and n-heptane (A0505-098-I1 and A0505-098-J1, Table 13) where no exchange took place, presumably because of the poor solubility of the solvate in these solvents.
Evidently, apomorphine•1*IPA Form A can alter its solvated composition if certain solvents or water are present at high enough activities to effect displacement. Partial replacement of IPA by the competitive solvent avoids disruption to the crystal phase by XRPD. However, solvates or hemi-solvates obtained from the suspension equilibration screen, in which complete replacement of the IPA occurred, resulted in new diffraction patterns by XRPD.
Precipitation of apomorphine•1*IPA from IPA, by isopropanol generated a product (A0505-098-N1, Table 13) that was consistent with the authentic starting material by XRPD. This supports the assumption that undesirable forms of apomorphine•1*IPA will not be generated by uncontrolled crystallization during manufacture, or if they are, they readily convert to or revert back into Form A.
Four suspension equilibrations (A0505-116-A1 to D1) were performed to include different combinations of input forms (Table 14). The mixtures were stirred in IPA (10.0 vol) at 45-50° C. for 6 days. The resulting solids were isolated by filtration, washed with recycled maturation solvent, dried under a stream of nitrogen at 18-23° C. and analyzed by XRPD. The diffraction patterns and isopropanol contents of the products from the suspension equilibrations were consistent with authentic apomorphine•1*IPA Form A.
After single forms or mixtures of single forms were suspended in anhydrous isopropanol at 45-50° C. and stirred for up to 6 days, the following observations were made:
These findings suggest that if other forms are generated in situ they can be readily controlled by suspension equilibration treatment with IPA.
Experiments were performed on samples of apomorphine•1*IPA (demonstration batch A0526-004-B1, after single crystallization), apomorphine•1*IPA (demonstration batch A0526-010-A1, after recrystallization) an alternative solvate apomorphine•0.5*EtOH (A0505-080-G1) and amorphous apomorphine, prepared via the aqueous route (A0530-020-01). All samples were maintained under a single humidity condition of 75 to 80% RH at 18-23° C. for 144 h. Uppermost areas of exposure of the samples were the same for all solids and the results are summarized in Table 15.
Water uptake of the apomorphine solvates was insignificant after 144 h, whilst the weight increase of amorphous apomorphine (A0486-080-C1) was 2.0% w/w after 1 h and had stabilized at this level after completion of the investigation. Uniform equilibration was assumed and all specimens were mobile after this time and appeared physically unaltered when observed. Their compositions by 1H NMR and form by XRPD were unchanged.
Apomorphine•1*IPA Form A (A0526-010-A1) was subjected to a step profile of 0 to 90% RH in 10% RH increments followed by a desorption profile of 85% RH to 0% RH in 10% RH decrements and the temperature was maintained at 25±0.1° C. The weight changes during the sorption/desorption cycle were monitored.
From 0% to 90% RH a weight change of <0.1% w/w was observed attributed to absorption of surface water, not bonded, this water was lost in desorption. The absorption profile showed that the sample was not hygroscopic according to European Pharmacopoeia classification of equilibration of the API (Table 16). Form changes were not observed.
The following solvates were identified: formamide, acetone, TBME, methyl acetate, THF, ethanol, acetonitrile, 2-propanol (solvate investigated in the polymorph screen), water, 1,4-dioxane, nitromethane, pyridine, and ethylene glycol. The solvates contained variable amounts of amorphous apomorphine free base. The diffraction patterns and proposed onset temperatures of solvent release for each of the solvates were measured and are reported in Example 14 and the corresponding figures.
Five crystalline solvates were unintentionally generated during the polymorph screen. Characterization data for these appears in Example 14 and the corresponding figures.
Apomorphine•1*IPA Form A is advantageous because it is easily prepared and the physical form is well controlled by crystallization. No alternative apomorphine•1*IPA polymorphic forms were identified during the polymorph screen and the solvate exhibited good resilience to elevated humidity conditions.
Some solvent exchange of IPA occurred when other solvents were present at high concentrations, but this can be avoided during the final recrystallization and isolation from IPA by employing IPA of sufficiently high grade (e.g., INEOS) pharm grade with a purity of 99.96% w/w and containing a low level of water (0.1% w/w).
DSC.
A Mettler Toledo DSC 821 instrument was used for the thermal analysis operating with STARe™ software. The analysis was conducted in 40 μl open aluminium pans, under nitrogen and sample sizes ranged from 1 to 10 mg. Typical analysis method was 20 to 350 at 10° C./minute.
FTIR.
FTIR Spectra were acquired using a PerkinElmer Spectrum One FTIR spectrometer. Samples were analyzed directly using a universal ATR attachment in the frequency range 4000 to 600 cm−1. Spectra were processed using Spectrum CFD, vs. 4.0 PerkinElmer Instruments LLC.
GVS.
The sample (approximately 7 mg) was placed into a wire mesh vapour sorption balance pan and loaded inside a Hiden Analytical Instruments IGAsorp vapour sorption balance and maintained at 25±0.1° C. The sample was then subjected to s step profile from 0 to 90% RH in 10% increments and then a desorption profile from 85% to 0% RH in 10% decrements. The weight change during the sorption cycle was monitored, allowing the hygroscopic nature of the sample to be determined.
1H NMR.
1H NMR Spectra were acquired using a Bruker 400 MHz spectrometer and data was processed using Topspin. 1H NMR Samples were prepared in DMSO-d6 and referenced to the non-deuterated solvent residual at 2.50 ppm.
Internal standard TCNB (30.8 mg, F.W. 260.89, 99%) and apomorphine (A0526-010-A1) (30.3 mg, F.W. 267.33) were dissolved in CD3OD (3.0 ml) and the 1H NMR spectrum was acquired using an extended relaxation (20 s) method (
Signal at δ=8.4 ppm (1H, d) corresponded to an aryl signal associated with the product (∫1.00). Signal at δ=8.5 ppm (s) corresponded to the internal standard TCNB (∫1.27).
METCR2498
Column: Hypersil BDS C18, 150×4.6 mm, 5 μm
Inj. volume: 10 μl
Detection: Ultra violet @ 280 nm
Mobile phase A: Sodium octanesulfonate, pH2.2
Mobile phase B: Acetonitrile
Gradient:
Flow rate: 1.5 ml/min
Column temperature: 35° C.
Run time: 50 minutes
Integration time: 37 minutes
Wash vial: Water/acetonitrile, 1/1 v/v
To prepare 2 L mobile phase A, weigh 2.2 g of sodium octanesulfonate monohydrate into a 2 L duran and dissolve in 2000 ml of deionized water. Adjust to pH 2.2 (±0.1) using a dilute orthophosphoric acid solution (1:1 with deionised water), mix well and degas by sonication.
To prepare 1 L mobile phase B, transfer 1000 ml acetonitrile to a 1 L mobile phase reservoir and degas by sonication.
To prepare 500 ml sample diluent transfer 500 ml deionized water to a 500 ml mobile phase duran. Add 5 ml of acetic acid. Mix well and degas by sonication.
Different volumes of mobile phase and/or sample diluent may be prepared as long as the proportions of each component remain the same.
Routine LC-MS data were collected using a Micro Mass platform LCZ interfaced with: CTC Analytics liquid sample changer system, Waters 2487 dual λ absorbance detector and Agilent series 1100 binary pump.
The instrument used a ZMD quadrupole mass analyser based detector and the mass separated ions were detected via a photomultiplier system. The ZMD quadrupole instrument was calibrated up to 2000 Da.
The instrument used for digital capture was an Olympus BX41 microscope with digital camera attachment. The magnification was ×100 and ×400. Samples were observed under plane-polarized and cross-polarized light.
TGA:A Perkin Elmer Pyris Diamond TG/DTA 6300 was used to measure the weight loss as a function of temperature from 30 to 600° C. The scan rate was 10° C./min and the purge gas was nitrogen.
HSM: The instrument used for digital capture was an Olympus BX41 microscope with digital camera and hot stage attachment. The magnification was ×100 and ×400. Samples were observed under plane-polarized and cross-polarized light.
XRPD analysis was carried out using a Bruker D2 Phaser powder diffractometer equipped with a LynxEye detector. The specimens underwent minimum preparation but, if necessary they were lightly milled in a pestle and mortar before acquisition. The specimens were located at the center of a silicon sample holder within a 5 mm pocket (approximately 5-10 mg).
The samples were continuously spun during data collection and scanned using a step size of 0.02° two theta (20) between the range of 4° to 40° two theta. Data was acquired using either 3 minute or 20 minute acquisition methods. Data was processed using Bruker Diffrac.Suite.
Scheme 7 summarizes the steps performed in the optimized synthesis. Each step is described in more detail below:
Output: 605 g, 85% th corrected for 18.9% w/w IPA assay against TCNB internal standard (A0526-004-B1, 99.4% w/w on anhydrous solvent free basis).
Photomicrograph: prisms (
XRPD: sharp diffraction peaks, consistent with previous batches (
Recrystallization (A0526-010) Note: All manipulations were performed under nitrogen.
Output (A0526-010-A1): 503 g, 91% th (100.7% w/w on anhydrous solvent free basis, 19.2% w/w IPA, by 1H NMR), assay against TCNB internal standard.
XRPD: sharp diffraction peaks, consistent with previous batches (
QC analysis: see
Scheme 8 summarizes the steps performed in the optimized aqueous-based procedure. Each step is described in more detail below:
The following solvates were identified: formamide, acetone, TBME, methyl acetate, THF, ethanol, acetonitrile, 2-propanol (solvate investigated in the polymorph screen), water, 1,4-dioxane, nitromethane, pyridine, and ethylene glycol. The solvates contained variable amounts of amorphous apomorphine free base. The diffraction patterns and proposed onset temperatures of solvent release for each of the solvates were measured and are reported below.
2Theta: 7.489, 7.588, 8.192, 9.130, 10.978, 12.232, 13.529, 14.037, 14.928, 19.569, 20.241, 20.706, 21.859, 22.547, 22.898, 23.328, 24.066, 24.307, 25.313, 26.047, 26.834, 29.855, 33.007.
2Theta: 7.626, 8.780, 9.443, 10.327, 12.709, 13.053, 13.986, 14.821, 15.534, 16.590, 17.130, 17.621, 18.849, 19.610, 18.358, 20.273, 21.010, 22.729, 23.122, 23.392, 24.227, 26.806, 25.503, 28.535, 29.441, 30.622.
2Theta: 8.289, 10.534, 14.010, 14.927, 18.727, 20.048, 22.104.
2Theta: 9.085, 10.615, 11.442, 12.771, 13.172, 13.899, 14.689, 15.291, 16.883, 18.195, 18.486, 19.028, 19.570, 20.629, 21.316, 21.647, 23.009, 24.239, 25.906, 26.858, 27.850, 29.163, 33.626, 35.225.
2Theta: 10.623, 10.905, 11.891, 12.572, 13.136, 13.840, 14.779, 16.212, 17.292, 17.808, 18.808, 19.922, 21.213, 21.847, 22.833, 24.266, 27.412, 29.007.
2Theta: 10.585, 11.980, 12.768, 13.091, 14.344, 14.526, 15.596, 15.960, 17.637, 18.446, 18.708, 19.678, 20.224, 20.689, 21.497, 22.467, 24.326, 25.437, 26.387, 27.577, 28.067, 32.313, 28.850, 24.036.
2Theta: 5.817, 9.377, 10.668, 11.655, 12.125, 13.161, 12.795, 14.150, 14.449, 14.671, 15.752, 17.289, 18.242, 19.259, 19.823, 20.266, 21.602, 22.852, 24.344, 25.423, 26.538, 27.547, 29.244.
2Theta: 7.593, 7.988, 8.383, 10.306, 11.305, 11.946, 12.423, 12.997, 13.397, 14.587, 16.101, 16.614, 17.128, 17.441, 17.883, 19.152, 19.716, 20.610, 22.272, 23.872, 25.025, 26.199, 27.160, 27.887, 28.535.
2Theta: 8.010, 8.779, 10.552, 11.038, 13.210, 14.057, 14.797, 14.962, 15.979, 16.816, 17.625, 18.398, 18.926, 19.478, 20.407, 21.896, 22.690, 22.996, 23.746, 24.197, 25.362, 26.374, 26.775, 27.255.
2Theta: 9.033, 10.633, 11.565, 16.594, 20.748, 21.173, 22.987, 23.652, 24.487.
2Theta: 11.731, 13.713, 14.859, 15.135, 18.047, 19.470, 21.359, 23.508, 24.428, 25.997, 29.428.
2Theta: 8.023, 10.511, 12.063, 12.899, 14.382, 14.850, 15.487, 15.925, 17.334, 18.353, 18.623, 20.078, 20.258, 20.767, 21.277, 22.049, 22.746, 23.937, 24.409, 24.975, 25.929, 26.790, 27.540, 27.874, 28.911, 29.865.
Solvates from the Polymorph Screen
2Theta: 8.194, 10.652, 11.468, 12.344, 14.096, 14.464, 15.493, 15.739, 16.392, 18.107, 18.404, 19.499, 20.025, 20.967, 21.502, 21.824, 22.359, 24.332, 24.766, 25.719, 26.455, 26.923, 28.444, 31.670, 32.289, 29.084.
2Theta: 8.282, 10.529, 13.709, 13.998, 14.892, 16.521, 17.772, 18.712, 18.953, 20.067, 20.907, 22.105, 23.789, 24.712, 25.758, 26.575, 27.700, 28.807.
2Theta: 7.953, 8.440, 11.182, 12.017, 12.754, 12.917, 15.942, 16.872, 17.338, 17.815, 20.374, 21.456, 23.176, 23.721, 25.627, 27.141, 24.280.
2Theta: 7.962, 10.599, 11.952, 12.778, 14.352, 14.527, 15.608, 15.925, 17.584, 18.375, 18.693, 19.688, 20.249, 20.668, 21.480, 22.159, 22.447, 24.024, 24.365, 25.404, 25.662, 26.428, 27.535, 28.036, 28.896, 29.360, 29.860, 30.262, 31.018, 32.308.
2Theta: 8.050, 10.387, 11.260, 12.351, 14.123, 15.253, 16.126, 17.808, 19.671, 19.945, 20.544, 21.211, 21.741, 24.341, 24.871, 25.247, 26.496, 27.967.
Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about”. Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention.
The entire contents of all patents, published patent applications, websites, and other references cited herein are hereby expressly incorporated herein in their entireties by reference.
This application claims the benefit of U.S. Provisional Patent Application No. 62/096,352, filed Dec. 23, 2014, and U.S. Provisional Patent Application No. 62/240,611, filed Oct. 13, 2015, the entire content of which being herewith incorporated by reference as if fully disclosed herein.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/IL2015/051246 | 12/23/2015 | WO | 00 |
| Number | Date | Country | |
|---|---|---|---|
| 62240611 | Oct 2015 | US | |
| 62096352 | Dec 2014 | US |
