HK1174261A - Co-crystals comprising vx-950 and pharaceutical compositions comprising the same - Google Patents

Description

Co-crystals comprising VX-950 and pharmaceutical compositions comprising the same

The present application is a divisional application of an invention patent application having an application date of 27/2/2007 (PCT/US2007/004995), application number 200780015244.5(PCT/US2007/004995), entitled "co-crystal comprising VX-950 and pharmaceutical composition comprising the co-crystal".

Cross-referencing

This application claims the benefit of U.S. provisional application 60/777,221 filed on 27.2006, month 2, which is incorporated herein by reference.

Background

Hepatitis c virus ("HCV") infection is a medical problem that humans must face. It is now recognized that HCV is the causative agent in most cases of non-a, non-b hepatitis. The global infection population is estimated to be 3% [ A. Alberti et al, "Natural History of Hepatitis C", J.hepatology, 31(suppl.1), pp.17-24 (1999) ]. Nearly four million people are infected in The United States alone [ M.J. Alter et al, "The epidemic of Viral Hepatitis in The United States," gastroenterol. Clin.North Am., 23, 437-455 (1994); alter, "Hepatitis C Virus Infection in the United States (Hepatitis C Virus Infection)", J.hepatology, 31(suppl.1), pp.88-91 (1999) ].

Only about 20% of infected individuals suffer from acute clinical hepatitis when initially exposed to HCV, while others appear to resolve the infection spontaneously. However, in almost 70% of cases, The virus establishes a Chronic infection that lasts for decades [ s.iwarson, "The natural course of Chronic Hepatitis", femmichiology Reviews, 14, pages 201-204 (1994); lavanchy, "globalservatillonance and Control of Hepatitis C (global monitoring and Control of Hepatitis C)", j.viral Hepatitis, pages 6, 35-47 (1999) ]. This often results in recurrent, progressive malignant Hepatitis, which will often lead to more severe conditions such as cirrhosis and liver cancer [ m.c. kew, "Hepatitis C and Hepatocellular cancer (Hepatitis C and liver cancer)", FEMS Microbiology Reviews, pages 14, 211-220 (1994); saito et al, "Hepatitis C Virus Infection Associated with the Development of Hepatitis C viral Carcinoma," Proc. Natl. Acad. Sci. USA, 87, 6547-6549 (1990). Unfortunately, there is no widely effective treatment for this slow progression of chronic HCV.

[041] The HCV genome encodes a 3010-3033 amino acid polyprotein [ Q.L.Choo et al, "Genetic Organization and Diversity of the Hepatitis C Virus (Genetic Organization and Diversity of Hepatitis C Virus)", Proc.Natl.Acad.Sci.USA, 88, pp.2451-2455 (1991); kato et al, "Molecular Cloning of the HumanHepatii s C Virus Genome From Japanese hepatitis Patients with Non-A, Non-BHEPTITIS (Molecular Cloning of the human hepatitis C Virus Genome From Japanese Non-A, Non-B hepatitis Patients)", Proc.Natl.Acad.Sci., USA, 87, 9524-9528 (1990); takamizawa et al, "Structure and Organization of the Hepatitis C Virus genome Isolated From Human Cariers (Structure and Organization of the Hepatitis C Virus genome Isolated From Human vectors)", J.Virol., 65, pp.1105-1113 (1991) ]. Presumably, the essential catalytic mechanism for viral replication is provided by the HCV Nonstructural (NS) proteins. The NS Protein is derived from the proteolytic Cleavage of polyproteins [ R.Bartenschlager et al, "non structural Protein 3 of the Hepatitis C Virus Encodes a series-type protease Required for Cleavage of NS3/4 and NS4/5Junctions (non structural Protein 3 of Hepatitis C Virus Encodes a Serine-type protease Required for Cleavage of NS3/4 and NS4/5 Junctions) ], J.Virol., 67, pages 3835-3844 (1993); grakoui et al, "Characterization of the Hepatitis C Virus-Encoded Serinoprotein: determination of protease-Dependent polyprotein cleavage Sites (characterization of hepatitis C-encoding serine proteases: Determination of protease-Dependent polyprotein cleavage Sites), "J.Virol., 67, pages 2832-2843 (1993); grakoui et al, "Expression and Identification of Hepatitis C Virus viral protein Cleavage Products (Expression and Identification of Hepatitis C virus polyprotein Cleavage Products)", J.Virol., 67, pages 1385-1395 (1993); tomei et al, "NS 3 isa serine protease required for processing of hepatitis C virus polyprotein (NS3 is a serine protease required for processing of hepatitis C virus polyprotein)", J.Virol., 67, pp.4017-4026 (1993) ].

HCV NS protein 3(NS3) is essential for viral replication and infectivity [ Kolykhalov, J.virology, Vol.74, pp.2046-2051 "Mutations of hepatitis atthe HCV NS3 spring Protease Catalytic triple infectious immunity of HCV RNA in Chimpanzees (Mutations of the HCV NS3 Serine Protease Catalytic Triad antiinfectivity of chimpanzee HCV RNA)" ]. It is known that NS3 Protease mutations in Yellow Fever Virus reduce viral infectivity [ Chambers, T.J. et al, "Evidence of an evolution of the N-tertiary domain of the Nonstructural Protein NS3 From Yellow river Virus viral Protein kinase response for Site-Specific cleavage of the N-terminal domain of the Nonstructural Protein NS3 From Yellow Fever Virus" Protein. The first 181 amino acids of NS3 (residues 1027-1207 of the viral polyprotein) appear to contain the Serine protease domain of NS3, which has all four downstream positions of the HCV polyprotein [ c.lin et al, "Hepatitis C Virus NS3 spring protease: trans-clean Requirements and Processing tools (hepatitis C NS3 serine protease: requirement and process Kinetics for Trans-Cleavage) ", J.Virol., 68, pp.8147-8157 (1994) ].

The HCV NS3 serine protease and its related cofactor NS4A help process all viral enzymes and are therefore considered essential for viral replication. Its processing appears to be similar to that of human immunodeficiency virus aspartyl protease (also involving viral enzyme processing). HIV protease inhibitors that inhibit viral protein processing are effective antiviral agents in humans, indicating that interrupting this phase of the viral life cycle results in therapeutically active agents. Therefore, the HCV NS3 serine protease is also an attractive target for drug discovery.

Until recently, the only therapy established for HCV disease was interferon treatment. However, interferons have significant side effects [ m.a. wlaker et al, "heparis CVirus: an Overview of Current applications and Progress (review of hepatitis C: Current methods and Progress), "DDT, pages 4, 518-29 (1999); moradpour et al, "Current and eventing therapeutics for Hepatitis C (Current and future treatment of Hepatitis C)", eur.j. gastroenterol.hepatol.11, pages 1199-1202 (1999); janssen et al, "Suicide association with alfa-interference Therapy for Viral Hepatitis" Suicide behavior accompanying Interferon alpha Therapy for Chronic Viral Hepatitis, j.hepatol., 21, pp 241-243 (1994); renault et al, "Side Effects of Alpha Interferon", Seminirs in Liver Disease, 9, pp.273-277 (1989)]And only a small fraction (-25%) of cases were induced to achieve long-term remission [ O.Weiland, "Interferon therapy in viral Hepatitis C Infection (interferon therapy for Chronic Hepatitis C Infection)", FEMS microbial. Rev., 14, 279-288 (1994)]. Recently, polyethylene glycol type interferon (PEG-And) And ribavirin and interferon combination therapyThe good turnover rate is only slightly improved, and the side effect is only partially reduced. In addition, the prospect of an effective vaccine against HCV is uncertain.

Therefore, more effective anti-HCV therapeutic drugs are needed. Such inhibitors should have therapeutic potential as protease inhibitors, especially as serine protease inhibitors, more especially as HCV NS3 protease inhibitors. In particular, these compounds are useful as antiviral agents, particularly as anti-HCV agents.

The HCV inhibitor VX-950, having the structure shown below, is a desirable class of such compounds. VX-950 is described in PCT publication No. WO 02/18369, which is incorporated herein by reference in its entirety.

Summary of The Invention

The present invention relates generally to compositions comprising the HCV inhibitor VX-950 and a specific co-crystal precursor (CCF): in some cases, VX-950 and CCF can together form a crystalline composition, i.e., a co-crystal. Particular VX-950 co-crystals are advantageous compared to their free form because they have improved dissolution, higher water solubility, and higher solid state physical stability than amorphous VX-950 dispersions. Certain VX-950 co-crystals provide reduced dosage form volume (mass of the dose form) and therefore have a lower dosing burden (pill burden) because VX-950 co-crystals exhibit higher bulk density relative to amorphous forms. Additionally, VX-950 co-crystals provide manufacturing advantages over amorphous forms that require spray drying, melt extrusion, lyophilization, or precipitation.

In one aspect, the invention provides compositions comprising VX-950 and a CCF compound selected from Salicylic Acid (SA), 4-aminosalicylic acid (4-ASA) and Oxalic Acid (OA), respectively, as CCF. In one embodiment, VX-950 and CCF together are in a crystalline form in the composition.

In another aspect, the invention provides three VX-950 co-crystals, the three co-crystals comprising VX-950 and CCF, respectively. In particular, the first co-crystal comprises VX-950 and Salicylic Acid (SA) as CCF. In some embodiments, when CCF is SA, the X-ray powder diffraction (XRPD) spectrum of the co-crystal shows peaks at about 4.43, 7.63, 8.53, 9.63, 12.89, 14.83, and 16.29 at 2 Θ; and a Differential Scanning Calorimetry (DSC) thermogram showing melting points at about 137 ℃ and about 223 ℃. The second co-crystal comprises VX-950 and 4-aminosalicylic acid (4-ASA) as CCF. In some embodiments, when the CCF is 4-ASA, the XRPD spectrum of the co-crystal shows peaks at about 4.37, 7.57, 8.47, 9.59, 12.81, and 14.75 in 2 Θ; and its DSC thermogram shows a melting point at about 177 ℃. The third eutectic comprises VX-950 and Oxalic Acid (OA) as CCF. In some embodiments, when CCF is OA, the XRPD spectrum of this co-crystal shows peaks at about 4.65, 6.17, 9.63, 12.65, 14.91, and 28.97 in 2 θ. In some embodiments, the ratio of the number of molecules of VX-950 to CCF in a unit cell is from 0.2 to 5 (e.g., 1). In some embodiments, VX-950 and CCF are both in the solid state (e.g., crystalline) and are non-covalently bound (e.g., by hydrogen bonding).

In another aspect, the invention provides a co-crystal of formula (VX-950) m (CCF) n, wherein CCF is a co-crystal precursor selected from salicylic acid, 4-aminosalicylic acid, and oxalic acid, and m and n are independently integers from 1 to 5. In some embodiments, m and n are both 1.

In another aspect, the invention provides a co-crystal of VX-950 and CCF, wherein the CCF is solid at room temperature and the VX-950 and CCF interact through non-covalent bonds. In some embodiments, CCF is selected from salicylic acid, 4-aminosalicylic acid, and oxalic acid. In certain embodiments, the non-covalent interactions between VX-950 and CCF include hydrogen bonding and van der Waals interactions.

In another aspect, the invention provides a pharmaceutical composition comprising one of the three VX-950 co-crystals described above. In one embodiment, the pharmaceutical composition further comprises a diluent, solvent, excipient or carrier.

Another aspect of the invention provides a method of preparing a co-crystal of VX-950 and CCF selected from salicylic acid, 4-aminosalicylic acid, and oxalic acid. The method comprises the following steps: providing VX-950; providing a co-crystal precursor salicylic acid, 4-aminosalicylic acid, or oxalic acid; grinding, heating, co-subliming, co-melting or contacting VX-950 with the co-crystal precursor in solution under crystallization conditions to form a co-crystal in a solid phase; the co-crystal thus formed is then optionally isolated. In some embodiments, preparing a co-crystal of VX-950 and CCF comprises providing VX-950 and CCF in a molar ratio of about 10 to about 0.1.

In another aspect, the invention provides a method of modulating a chemical or physical property of interest (e.g., melting point, solubility, hygroscopicity, and bioavailability) of a co-crystal comprising VX-950 and a CCF selected from salicylic acid, 4-aminosalicylic acid, and oxalic acid. The method comprises the following steps: detecting a chemical or physical property of interest associated with VX-950 and the co-crystal; determining the mole fraction of VX-950 and co-crystal precursor that needs to be adjusted to obtain the relevant chemical or physical property; and producing a co-crystal having the determined mole fraction.

The compositions and co-crystals of the invention are useful for treating diseases associated with or associated with HCV. Thus, the invention also includes a method of treating such diseases comprising administering to a subject in need thereof a therapeutically effective amount of a co-crystal of the invention or a composition of the invention.

The compositions and co-crystals of the invention can also be seeded to produce other co-crystals comprising an active ingredient that may be the same as or different from VX-950, and a CCF that may also be the same as or different from salicylic acid, 4-aminosalicylic acid, and oxalic acid. For example, a small amount of the co-crystal of the invention may be placed in a solution containing the desired active ingredient and CCF, and the resulting mixture placed so that the additional co-crystal is able to form and grow in the presence of the co-crystal present.

In addition, the compositions and co-crystals of the present invention are useful as research tools. For example, the crystal structure of the co-crystal can be used for molecular modeling to identify other possible co-crystal forms. They can be used to study pharmacological properties (such as bioavailability, metabolism and efficacy).

Brief Description of Drawings

FIG. 1 shows a eutectic thermogravimetric analysis (TGA) profile of VX-950 and SA.

FIG. 2 shows a co-crystal TGA spectrum of VX-950 and 4-ASA.

FIG. 3 shows a co-crystal DSC thermogram of VX-950 and SA.

FIG. 4 shows a co-crystal DSC thermogram of VX-950 and 4-ASA.

FIG. 5 shows the XRPD pattern of a co-crystal of VX-950 and 4-ASA at room temperature, water (top) and 1% HPMC (bottom) for 6 hours.

FIG. 6 shows the co-crystal XRPD patterns of VX-950 and SA at room temperature in water for 1 hour (top), 2 hours (middle) and 6 hours (bottom).

Figure 7 shows the co-crystal XRPD patterns of VX-950 and SA at room temperature, 1 hour (middle), 2 hours (top) and 6 hours (bottom) in 1% HPMC.

FIG. 8 shows the co-crystal XRPD pattern between VX-950 and SA.

FIG. 9 shows the co-crystal XRPD pattern of VX-950 and 4-ASA.

FIG. 10 shows the co-crystal XRPD pattern of VX-950 and OA.

Detailed Description

Methods for preparing and characterizing co-crystals are well described in the literature. See, e.g., Trask et al, chem.Commun., 2004, 890-891; almarsson and m.j.zawortko, chem.commun., 2004, 1889-1896. These methods are also generally applicable to the preparation and characterization of the co-crystals of the present invention.

Examples of co-crystals prepared with the active pharmaceutical ingredient and CCF include ball milling, melting in a reaction block, evaporation of solvent, slurry conversion, blending, sublimation, or modeling. In the ball milling process, a molar ratio of the components of the co-crystal (e.g., the compounds of interest in the present invention such as VX-950 and CCF) are mixed and milled with the balls. A solvent, such as methyl ethyl ketone, is optionally added to the mixture being ball milled. After milling, the mixture may be dried at room temperature or under heating conditions under vacuum, typically to give a powder product. In the melt process, the components of the co-crystal (e.g., CCF and VX-950) are combined, optionally with a solvent (e.g., acetonitrile). The mixture was then placed in a reaction block with a lid and subsequently heated to an endothermic temperature. The resulting reaction mixture is then cooled and the solvent (if used) is removed. In the solvent evaporation method, the components of the co-crystal are first dissolved separately in a solvent (or solvent mixture, such as 50/50 toluene and acetonitrile) and the solutions are then mixed together. The mixture was then allowed to stand and the solvent evaporated to dryness to give a co-crystal.

[0032] Examples of characterization methods include thermogravimetric analysis (TGA), Differential Scanning Calorimetry (DSC), X-ray powder diffraction (XRPD), solubility analysis, dynamic vapor adsorption, infrared off-gas analysis (infra-red off-gas analysis), and suspension stability. TGA can be used to study the presence of residual solvent in the co-crystal samples and determine the temperature at which decomposition of each co-crystal sample occurs. DSC can be used to look for the thermal transitions that occur in the eutectic samples as a function of temperature and determine the melting point of each eutectic sample. The co-crystals can be structurally characterized by XRPD. Solubility analysis can be performed to reflect changes in the physical state of each cocrystal sample. Suspension stability analysis can also be used to determine the chemical stability of the co-crystal samples in solvent.

An effective amount of a co-crystal or composition of the invention comprising VX-950 and a co-crystal precursor (CCF) selected from salicylic acid, 4-aminosalicylic acid, and oxalic acid, respectively, can be used to treat a disease associated with or associated with HCV. An effective amount is that amount necessary to impart a therapeutic effect to the subject being treated (e.g., a patient). An effective amount of the co-crystal of VX-950 and CCF is about 0.1mg/kg to about 150mg/kg (e.g., about 1mg/kg to about 60 mg/kg). As will be recognized by those skilled in the art, effective dosages will also vary depending upon the route of administration, the use of excipients, and the possibility of co-use with other therapies, including the use of other therapeutic agents and/or therapies.

The co-crystal or pharmaceutical composition of the invention can be administered to a subject (e.g., a cell, tissue, or patient (including animal or human)) in need thereof by any method that allows for the delivery of the compound VX-950, such as orally, intravenously, or parenterally. For example, they can be used for ingestion or injection by pills, tablets, capsules, aerosols, suppositories, liquid preparations, or as eye drops or ear drops, food supplements and topical preparations.

The pharmaceutical compositions may contain diluents, solvents, excipients and carriers such as water, ringer's solution, isotonic saline, 5% dextrose and isotonic sodium chloride solution. In another embodiment, the pharmaceutical composition may further comprise a solubilizing agent, such as a cyclodextrin. Additional examples of suitable diluents, solvents, excipients, carriers and solubilizers can be found in, for example, U.S. pharmacopeia 23/National Formulary 18, Rockville, MD, U.S. pharmacopeia Convention, inc. (1995); ansel HC, Popovich NG, Allen Jr LV pharmaceutical document and Drug Delivery Systems, Baltimore MD, Williams & Wilkins, (1995); gennaro ar, Remingtons: the Science and Practice of Pharmacy, Easton PA, Mack publishing Co., (1995); wade a., Weller pj. handbook of pharmaceutical Excipients, second edition, Washington DC, american pharmaceutical Association (1994); baner GS, Rhodes CT.Modern pharmaceuticals, third edition, New York, Marcel Dekker, Inc. (1995); ranade VV, Hollinger MA. drug Delivery systems. boca Raton, CRCPress, (1996).

The pharmaceutical compositions may also comprise an aqueous solution of the co-crystal in isotonic saline, 5% glucose or other well-known pharmaceutically acceptable excipients. Solubilizing agents, such as cyclodextrins or other solubilizing agents familiar to those skilled in the art, can be used as pharmaceutical excipients for the delivery of the therapeutic compound VX-950. With respect to the route of administration, the co-crystal or pharmaceutical composition may be administered orally, intranasally, transdermally, intradermally, vaginally, intraotically, intraocularly, buccally, rectally, transmucosally, or by inhalation or intravenously. The composition may also be delivered intravenously via a balloon catheter. The composition can be administered to an animal (e.g., a mammal, e.g., a human, a non-human primate, a horse, a dog, a cow, a pig, a sheep, a goat, a cat, a mouse, a guinea pig, a rabbit, a hamster, a gerbil, a ferret, a lizard, a reptile, or a bird).

The co-crystals or pharmaceutical compositions of the invention may also be delivered by implantation (e.g., surgically) using an implantable device. Examples of implantable devices include, but are not limited to, stents, drug delivery pumps, vascular filters, and implantable controlled release compositions. Any implantable device can be used to deliver compound VX-950 as an active ingredient of a co-crystal or pharmaceutical composition of the invention, provided that 1) the device, compound VX-950, and any pharmaceutical composition comprising the compound are biocompatible, and 2) the device can deliver or release an effective amount of the compound to deliver a therapeutic effect to a patient being treated.

Delivery of therapeutic agents through stents, drug delivery pumps (e.g., mini-osmotic pumps), and other implantable devices is known in the art. See, e.g., Hofma et al, "recent developments in Coated Stents", current international pathology Reports, 2001, 3: 28-36, the entire contents of which, including the references cited therein, are incorporated herein by reference. Additional descriptions of implantable devices (e.g., stents) can be found in U.S. Pat. Nos. 6,569,195 and 6,322,847, and PCT International publications WO04/0044405, WO 04/0018228, WO 03/0229390, WO 03/0228346, WO 03/0225450, WO 03/0216699, and WO 03/0204168, each of which (and other publications cited therein) is incorporated herein by reference in its entirety.

Examples of the preparation and characterization of the co-crystals of the present invention are described below, and these examples are intended to be illustrative only and not to limit the invention in any way.

Example 1 preparation by ball milling method

Salicylic Acid (SA): 70mg VX-950 and an equimolar equivalent of SA as CCF (Sigma Chemicals Co. (Sigma chemical Co.), St. Louis, Mo., USA) were mixed with 50. mu.L of methyl ethyl ketone ("MEK"). These components were ground for 10 minutes using a Wig-L-Bug apparatus. After milling, one batch was dried in a vacuum oven at 75 ℃ for 2 hours. The material obtained was off-white.

4-Aminosalicylic acid (4-ASA): 70mg VX-950 and equimolar equivalents of 4-ASA (15.8mg) (Sigma Chemicals Co. (Sigma chemical Co., St. Louis, Mo., USA) as CCF were mixed with 50. mu.L of acetonitrile ("ACN"). These components were then milled for 3 hours with a ball milling apparatus Retsch MM200(glen mills Inc, Clifton, NJ) at a frequency of 15 Hz. The mixture was placed in a grinding compartment made of sintered corundum. After milling, the material was transferred to a 20mL screw-cap scintillation vial (without cap) and dried in vacuo at room temperature for 16 hours. After drying, the lid was screwed on. The material obtained was off-white.

Oxalic Acid (OA): 70mg VX-950 and equimolar equivalents OA as CCF (Sigma Chemicals Co. (Sigma chemical Co., St. Louis, Mo., USA) were mixed with 5. mu.L of at least one of the following solvents (calculated on 10mg of total solids): ethyl acetate, methyl ethyl ketone, acetonitrile, water or 1, 2-dichloroethane. The components were milled together. After carrying out the same procedure as described above, a co-crystal of VX-950 and OA is obtained.

Example 2 preparation by melt Process

100mg VX-950 and an equimolar equivalent of a CCF selected from salicylic acid, 4-aminosalicylic acid and oxalic acid (Sigma Chemicals Co. (Sigma chemical Co., St. Louis, Mo., USA) were vortex mixed for 5 minutes. This step was performed twice. Once in the absence of solvent. In a second run, 4-aminosalicylic acid, salicylic acid and oxalic acid were treated with 100 μ L of acetonitrile, methyl ethyl ketone and ethyl acetate, respectively. The mixture was placed in a reaction block (reaction block) with a lid (Radiy Discovery Technologies, RR98072) and heated to an endothermic heat. The mixture was held at the endothermic temperature for 30 minutes, then the resulting mixture was cooled under de-lidded ambient conditions and the solvent removed (if used).

Example 3 preparation by solvent Evaporation

VX-950 and a CCF selected from salicylic acid, 4-aminosalicylic acid and oxalic acid (sigma chemicals Co. (sigma chemical company), st.louis, MO, USA) were each dissolved in a solvent mixture of 50% toluene/acetonitrile. Dissolution was aided by spinning and sonication until the solution was visually clear. The VX-950 solution and CCF solution were mixed in a 20mL spiral-cap scintillation vial at 0: 1, 1: 3, 1: 1 and 3: 1, 1: 0 molar ratios to give a final volume of 3 mL. The tubes were uncapped into a fume hood and the solvent was evaporated to dryness over a period of days to give a solid material.

Example 4 preparation by modeling

Modeling also yielded a Co-crystal of VX-950 and a CCF selected from salicylic acid, 4-aminosalicylic acid and oxalic acid (Sigma Chemicals Co. (Sigma chemical company), st.

EXAMPLE 5 thermogravimetric analysis (TGA)

TGA testing of each sample was performed using a Q500thermogravimetric analyzer (TA Instruments, New Castle, DE, USA) with control of ThermalAdvantage Q Series^TMSoftware, version 2.2.0.248, Thermal AdvantageRelease 4.2.1(TA Instruments-Water LLC), with the following components: qadv.exe version 2.2build 248.0; rhdii.dll version 2.2build 248.0; rhbase.dll version 2.2build 248.0; rhcomm. dll version 2.2build 248.0; taliense.dll version 2.2build 248.0; and tga. dll version 2.2build 248.0. Furthermore, the Analysis software used was Universal Analysis 2000 software (operating System Windows 2000/XP, version 4.1D build4.1.0.16(TA Instruments).

For all experiments, the basic procedure for performing the TGA test involved moving an aliquot of the sample (about 3-8mg) into a platinum sample pan (pan: part number 952018.906, TAInstructions). The disks were placed on a loading station and then automatically loaded into a Q500Thermogravimetric Analyzer (Thermogravimetric Analyzer) using control software. Thermograms were obtained by heating samples alone under flowing dry nitrogen (compressed nitrogen, grade 4.8 (bocvas, Murray Hill, NJ, USA) at a ramp rate of 10 ℃/min over a range of temperatures (typically room temperature to 300 ℃), sample purge flow rate of 90L/min, equilibrium purge flow rate of 10L/min.

As shown in FIG. 1, the co-crystal TGA spectrum of VX-950 and SA (molar ratio 1) shows about 2.3% weight loss at 145 ℃ and 18% total weight loss at 160 ℃. This is consistent with the expected loss of salicylic acid from the 1: 1 cocrystal. The first weight loss was probably due to sublimation of salicylic acid, which began at 76 ℃.

As shown in fig. 2, the co-crystal TGA spectrum of VX-950 and 4-ASA (again, in a 1 molar ratio) shows about a 1.4% weight loss (due to solvent release) at 125 ℃ and about a 13% weight loss at 250 ℃.

Example 6 Differential Scanning Calorimetry (DSC)

DSC analysis was performed using MDSC Q100 Differential scanning calorimeterInstruments) (TA Instruments) using control ThermalAdvantage Q Series^TMThe software, version 2.2.0.248, Thermal AdvantageRelease 4.2.1, has the following components: qadv.exe version 2.2build 248.0; rhdii.dll version 2.2build 248.0; rhbase.dll version 2.2build 248.0; rhcomm. dll version 2.2build 248.0; taliense.dll version 2.2build 248.0; and dsc.dll version 2.2 built 248.0. Furthermore, the analysis software used was universal analysis 2000 software (operating system Windows 2000/XP, version 4.1D build4.1.0.16(TA Instruments).

For all DSC experiments, an aliquot of the sample (about 2mg) was weighed and transferred into an aluminum sample pan (pan: part number 900786.901; lid: part number 900779.901, TA Instruments). The sample discs were closed by crimping with a single well and then loaded into a Q100 Differential scanning calorimeter (Differential scanning calorimeter) equipped with an autosampler. Thermograms were obtained by heating each sample separately under flowing dry nitrogen (compressed nitrogen, grade 4.8 (BOC Gases, MurrayHill, NJ, USA), at a ramp rate of 10 ℃/min, over a range of temperatures (typically room temperature to 300 ℃), a sample purge flow rate of 60L/min, and an equilibrium purge flow rate of 40L/min.

As shown in fig. 3, the DSC thermogram shows that the co-crystal of VX-950 and SA melts first at about 137 ℃. The melting points of SA and VX-950 were 159 ℃ and 247 ℃, respectively. The second melt transition at 223 ℃ corresponds roughly to the melt transition of the free compound. The lower melting point was observed due to the presence of impurities, but may include some decomposition.

As shown in FIG. 4, the DSC thermogram shows that the co-crystal of VX-950 and 4-ASA melted at about 177 ℃.

Table 1 shows a summary of DSC screens of potential interactions between VX-950 and CCF used in the present invention.

TABLE 1

Example 7X-ray powder diffraction (XRPD)

In the XRPD analysis, instruments from Bruker or Rigaku were used.

a.Bruker

XRPD patterns were obtained in reflection mode at room temperature using a Bruker D8Discover diffractometer (Bruker AXS, Madison, WI, USA) equipped with a sealed tube source and a Hi-Star planar detector. A copper palladium X-ray tube (Siemens) was operated at 40kV and 35 mA. Parallel monochromatic beams were generated using a graphite monochromator and 0.5mm collimator supplied by Bruker (CuKa,). The distance between the sample and the detector was about 30 cm. The sample was placed on a Si zero background wafer (The Gem Dugout, State College, Pa.), and then on an XYZ stage, centered on The XYZ stage. Data were collected with GADDS software (operating system Windows NT, version 4.1.16) (Bruker AXS, Madison, WI, USA). Two frames were recorded at 120 seconds per frame exposure time. During exposure, the sample was vibrated in both the X and Y directions with an amplitude of 1 mm. The data was then integrated in 0.02 steps over the entire range of 3 to 41 in 2 theta and merged into one continuous pattern. The instrument was calibrated using a corundum plate (NIST standard 1976).

b.Rigaku

XRPD patterns were recorded in transmission mode at room temperature using a rotating anode RUH3R X-ray generator (Rigaku, the woodlands, TX, USA) and a Rigdm rasis IIC detector. CuK radiation at 50kV and 100mA was used. Generation of parallel monochromatic beams with focusing mirrors and Ni filtersDuring the experiment, the samples were held in a 2mm diameter boron glass capillary (Hampton Research, Aliso Viejo, CA, USA) and rotated about the f-axis. The distance between the sample and the detector was about 25 cm. A single frame of 300 second exposure time was recorded with crystalcear software developed by Rigaku, version 1.3.5SP 2. The data was then integrated in steps of about 0.02 deg. over a range of 3 deg. to 41 deg. at 2 q. The instrument was calibrated with silicon powder (NIST standard 640 c).

As shown in fig. 5, after 6 hours at room temperature, in water and in 1% Hydroxypropylmethylcellulose (HPMC), the co-crystal of VX-950 and 4-ASA showed no sign of conversion to the free form after up to 6 hours incubation time. At the 24 hour time point, the co-crystals remained intact in the 1% HPMC solution. However, the sample in water had converted back to the free form at the 24 hour time point.

In contrast, as shown in FIG. 6, the XRPD pattern of the co-crystal of VX-950 and SA after suspension in water at room temperature shows: (i) after 1 hour, the co-crystal showed a slight conversion to the free form as shown by the increase in the peak at 9.12-theta (°), (ii) at the 2 hour time point, showed additional conversion, and (iii) at the 6 hour time point, showed complete conversion. After suspension in 1% HPMC aqueous solution, the same co-crystals showed a slight conversion from the co-crystals to the free form at the 1 hour time point and the 2 hour time point, and an additional conversion was observed at the 6 hour time point. See fig. 7.

Fig. 8, 9 and 10 are XRPD spectra of VX-950 co-crystals of SA, 4-ASA and OA, respectively. Specifically, the XRPD of the VX-950 and SA co-crystal shows peaks at 4.43, 7.63, 8.53, 9.63, 12.89, 14.83, and 16.29 at 2 θ; XRPD of VX-950 and 4-ASA co-crystal showed peaks at 4.37, 7.57, 8.47, 9.59, 12.81, and 14.75 in 2 Θ; the XRPD of the VX-950 and OA co-crystal shows peaks at 4.65, 6.17, 8.21, 9.63, 12.65, 14.91 and 28.97 in 2 θ.

Example 8 solubility analysis

Placing an aliquot of the sample into a tube, and then adding waterA sexual medium. Aliquots of the supernatant were withdrawn at set time points, filtered through 0.45PTFE micron filters (Millex, LCR, Millipore) and processed for High Performance Liquid Chromatography (HPLC) analysis (Agilent 1100; Palo Alto, Calif., USA). The system was equipped with an autosampler set at 25 ℃. The sample was treated and an aliquot of the sample was diluted 1: 1 (by volume) with acetonitrile. The samples were tested using detector isocratically (isocratically) set at 270 nm. The column isPhenyl column 150 mm. times.4.6 mm, 3.5 μm particle size (P/N186001144) (Waters, Milford, MA, USA). The mobile phase was 60: 40(v/v) potassium phosphate buffer (10mM, pH 7.0) to methanol. The assay was run at a flow rate of 1mL/min and completed within 15 min. Table 2 below summarizes the solubility of VX-950 and its co-crystal with 4-ASA (expressed as VX-950 equivalents (VX-950 elutes at 8.8 minutes)) in simulated intestinal fluid (pH 6.8) at 24 hour time points, room temperature.

TABLE 2

Example 9 suspension stability

The physical stability of the co-crystals when suspended in an aqueous medium was evaluated. The co-crystal powder was slurried at 25 ℃ at a nominal concentration of about 6mg/ml in (1) unbuffered deionized water and (2) a 1% (w/w) HPMC solution (low viscosity grade). The slurry was mixed with a magnetic stir bar and plate. Solid samples were isolated by filtration at 1, 2, 6 and 24 hour intervals.

After suspension in water for 1, 2, and 6 hours, the XRPD pattern of the co-crystal of VX-950 and salicylic acid shows a slight conversion of the co-crystal to the free form after 1 hour, as shown by the increase in the peak at 9.12 θ (°). Additional conversion was observed at the 2 hour time point and complete conversion was found at the 6 hour time point.

XRPD patterns of co-crystals of VX-950 and salicylic acid after suspension in 1% HPMC in water for 1, 2, and 6 hours showed slight conversion of the co-crystals to the free form at the 1 hour time point and the 2 hour time point. Additional conversion was observed at the 6 hour time point. HPMC appears to have reduced the rate of conversion of co-crystals to the free form. The slow transition is also evidenced by a rising peak at 9.12 θ (°).

Co-crystal XRPD pattern of VX-950 and 4-aminosalicylic acid after 6 hours of suspension in water and 1% HPMC in water. In both cases, the co-crystals showed no sign of conversion to the free form after up to 6 hours incubation time. At the 24 hour time point, the co-crystals remained intact in the 1% HPMC solution. However, the sample in water had converted back to the free form at the 24 hour time point.

Other embodiments

It is to be understood that while the specification has described the invention in detail, the foregoing detailed description is intended to illustrate and not limit the scope of the invention, which is defined in the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims (20)

1. A co-crystal comprising VX-950 and salicylic acid.

2. The co-crystal of claim 1, wherein the molar ratio of VX-950 and salicylic acid is about 1: 1.

3. The co-crystal of claim 2, having an X-ray powder diffraction peak, in terms of 2 Θ, at about 4.43, 7.63, 8.53, 9.63, 12.89, 14.83, 16.29.

4. The co-crystal of claim 2 having a DSC thermogram with peaks at about 137 ℃ and 223 ℃.

5. A co-crystal comprising VX-950 and 4-aminosalicylic acid.

6. The co-crystal of claim 5, wherein the molar ratio of VX-950 and 4-aminosalicylic acid is about 1: 1.

7. The co-crystal of claim 6, having X-ray powder diffraction peaks, in terms of 2 θ, at about 4.37, 7.57, 8.47, 9.59, 12.81, and 14.75.

8. The co-crystal of claim 6, having a DSC thermogram with a peak at about 177 ℃.

9. A co-crystal comprising VX-950 and oxalic acid.

10. The co-crystal of claim 9, wherein the molar ratio of VX-950 and oxalic acid is about 1: 1.

11. The co-crystal of claim 10, having X-ray powder diffraction peaks, in terms of 2 Θ, at about 4.65, 6.17, 8.21, 9.63, 12.65, 14.91, and 28.97.

12. One type (VX-950)_m:(CCF)_nWherein CCF is a co-crystal precursor selected from salicylic acid, 4-aminosalicylic acid and oxalic acid, and m and n are independently integers from 1 to 5.

13. The co-crystal of claim 12, wherein m and n are both 1.

14. A pharmaceutical composition comprising VX-950 and a co-crystal former selected from the group consisting of salicylic acid, 4-aminosalicylic acid, and oxalic acid.

15. The pharmaceutical composition of claim 14, wherein VX-950 and the co-crystal former are together in crystalline form.

16. The pharmaceutical composition of claim 14, wherein the molar ratio of VX-950 and the co-crystal precursor is about 1.

17. The pharmaceutical composition of claim 15, further comprising a diluent, solvent, excipient, carrier, or solubilizing agent.

18. A method of making a co-crystal, the method comprising:

a. providing a solution of VX-950 in a solvent,

b. providing a co-crystal precursor selected from salicylic acid, 4-aminosalicylic acid and oxalic acid,

c. grinding, heating, co-subliming, co-melting or contacting VX-950 with a co-crystal precursor in solution under crystallization conditions to form a co-crystal in a solid phase, and

d. optionally isolating the co-crystal formed in step (c).

19. A method of modulating a chemical or physical property of interest of a co-crystal, the method comprising:

a. detecting the relevant chemical or physical properties of VX-950 and the co-crystal precursor,

b. determining the mole fraction of VX-950 and co-crystal precursor that will give the desired modulation of the relevant chemical or physical property, and

c. preparing a co-crystal having the mole fraction determined in step (b).

20. A method of making a co-crystal, the method comprising providing an existing co-crystal as a seed for making a co-crystal, wherein the existing co-crystal comprises VX-950 and a co-crystal precursor selected from the group consisting of salicylic acid, 4-aminosalicylic acid, and oxalic acid; and the co-crystal comprises an active ingredient that may be the same as or different from VX-950, and also a co-crystal precursor that may be the same as or different from the co-crystal precursor contained in the existing co-crystal.