WO2014055069A1

WO2014055069A1 - Solid forms comprising an inhibitor of hcv ns5a, compositions thereof, and uses therewith

Info

Publication number: WO2014055069A1
Application number: PCT/US2012/058503
Authority: WO
Inventors: Leping Li; Anna Muchnik; Min Zhong; Nathan C. SCHULTHEISS
Original assignee: Presidio Pharmaceuticals, Inc.
Priority date: 2012-10-02
Filing date: 2012-10-02
Publication date: 2014-04-10

Abstract

Solid forms comprising methyl N-[(2S)-1-[(2S)-2-{5-[4-(2-{2-[(2S)-1-[(2S)-2- [(methoxycarbonyl)amino] -3 -methylbutanoyl]pyrrolidin-2-yl]- 1H-1,3 -benzodiazol-6- yl}ethynyl)phenyl]- 1H-imidazol-2-yl}pyrrolidin-1-yl] -3 -methyl-1-oxobutan-2-yl] carbamate dihydrochloride, compositions comprising the solid forms, methods of making the solid forms, and methods of their use for the treatment of HCV infection are disclosed.

Description

SOLID FORMS COMPRISING AN INHIBITOR OF HCV NS5A. COMPOSITIONS

THEREOF, AND USES THEREWITH

1. FIELD

[0001] Provided herein are solid forms comprising the compound of formula (I), compositions comprising the solid forms, methods of making the solid forms, and methods of their use in inhibiting hepatitis C virus ("HCV") replication, including, for example, functions of the non-structural 5A ("NS5A") protein of HCV.

(I)

2. BACKGROUND

(a) Hepatitis C Virus

[0002] HCV is a single-stranded RNA virus that is a member of the Flaviviridae family. The virus shows extensive genetic heterogeneity as there are currently seven identified genotypes and more than 50 identified subtypes. In HCV infected cells, viral RNA is translated into a polyprotein that is cleaved into ten individual proteins. At the amino terminus are structural proteins: the core (C) protein and the envelope glycoproteins, El and E2. p7, an integral membrane protein, follows El and E2. Additionally, there are six nonstructural proteins, NS2, NS3, NS4A, NS4B, NS5A and NS5B, which play a functional role in the HCV lifecycle. (see, for example, Lindenbach, B.D. and CM. Rice, Nature.

(2005) 436:933-938).

[0003] Infection by HCV is a serious health issue. It is estimated that 170 million people worldwide are chronically infected with HCV. HCV infection can lead to chronic hepatitis, cirrhosis, liver failure and hepatocellular carcinoma. Chronic HCV infection is thus a major worldwide cause of liver-related premature mortality.

[0004] The present standard of care treatment regimen for chronic HCV infection involves interferon-alpha, alone, or in combination with ribavirin. The treatment is cumbersome and sometimes has debilitating and severe side effects and many patients do not durably respond to treatment. New and effective methods of treating HCV infection, including for example, chronic HCV infection, are urgently needed.

[0005] Essential features of the NS5 A protein of HCV make it an ideal target for inhibitors.

(b) Solid Forms

The preparation and selection of a solid form of a pharmaceutical compound is complex, given that a change in solid form may affect a variety of physical and chemical properties, which may provide benefits or drawbacks in processing, formulation, stability and

bioavailability, among other important pharmaceutical characteristics. Potential

pharmaceutical solids include crystalline solids and amorphous solids. Amorphous solids are characterized by a lack of long-range structural order, whereas crystalline solids are characterized by structural periodicity. The desired class of pharmaceutical solid depends upon the specific application; amorphous solids are sometimes selected on the basis of, e.g., an enhanced dissolution profile, while crystalline solids may be desirable for properties such as, e.g., physical or chemical stability (see, e.g., S. R. Vippagunta et ah, Adv. Drug. Deliv. Rev., (2001) 48:3-26; L. Yu, Adv. Drug. Deliv. Rev., (2001) 48:27-42).

[0006] Whether crystalline or amorphous, potential solid forms of a pharmaceutical compound may include single-component and multiple-component solids. Single-component solids consist essentially of the pharmaceutical compound in the absence of other

compounds. Variety among single-component crystalline materials may potentially arise from the phenomenon of polymorphism, wherein multiple three-dimensional arrangements exist for a particular pharmaceutical compound (see, e.g., S. R. Byrn et ah, Solid State Chemistry of Drugs, (1999) SSCI, West Lafayette).

[0007] Additional diversity among the potential solid forms of a pharmaceutical compound may arise from the possibility of multiple-component solids. Crystalline solids comprising two or more ionic species are termed salts (see, e.g. , Handbook of Pharmaceutical Salts: Properties. Selection and Use. P. H. Stahl and C. G. Wermuth, Eds., (2002), Wiley, Weinheim). Additional types of multiple-component solids that may potentially offer other property improvements for a pharmaceutical compound or salt thereof include, e.g., hydrates, solvates, co-crystals and clathrates, among others (see, e.g., S. R. Byrn et ah, Solid State Chemistry of Drugs, (1999) SSCI, West Lafayette). Moreover, multiple-component crystal forms may potentially be susceptible to polymorphism, wherein a given multiple-component composition may exist in more than one three-dimensional crystalline arrangement. The discovery of solid forms is of great importance in the development of a safe, effective, stable and marketable pharmaceutical compound.

3. SUMMARY

[0008] Embodiments herein provide solid forms of the compound of formula (I)

("Compound (I)"):

[0009] Without intending to be limited by any particular theory, the storage stability, compressibility, bulk density or dissolution properties of certain solid forms described herein are believed to be beneficial for manufacturing, formulation and bioavailability of the compound of Formula (I).

[0010] The solid forms provided herein are useful as active pharmaceutical ingredients for the preparation of formulations for use in animals or humans. Thus, embodiments herein encompass the use of these solid forms as a final drug product. Certain embodiments provide solid forms useful in making final dosage forms with improved properties, e.g., powder flow properties, compaction properties, tableting properties, stability properties, and excipient compatibility properties, among others, that are needed for manufacturing, processing, formulation and/or storage of final drug products. Certain embodiments herein provide pharmaceutical compositions comprising a single-component crystal form, a multiple- component crystal form, a single-component amorphous form and/or a multiple-component amorphous form comprising the compound of formula (I) and a pharmaceutically acceptable diluent, excipient or carrier. The solid forms described herein are useful, for example, for inhibiting HCV replication, inhibiting NS5 A, and treating, preventing or managing HCV infection.

4. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] FIG. 1 provides a representative X-ray powder diffraction (XRPD) pattern of Form LC-B of Compound (I). [0012] FIG. 2 provides a representative XRPD pattern for Form B of Compound (I).

[0013] FIG. 3 provides a representative differential scanning calorimetry (DSC) thermogram of Form B of Compound (I).

[0014] FIG. 4 provides a representative thermal gravimetric analysis (TGA) thermogram of Form B of Compound (I).

[0015] FIG. 5 provides a representative XRPD pattern for Form A of Compound (I).

[0016] FIG. 6 provides a representative DSC thermogram of Form A of Compound (I).

[0017] FIG. 7 provides a representative TGA thermogram of Form A of Compound (I).

[0018] FIG. 8 provides a representative 1H NMR spectrum of a sample of Form A dissolved in DMSO-d6.

[0019] FIG. 9 provides a representative dynamic vapor sorption/desorption curve for Form LC-B of Compound (I).

[0020] FIG. 10 provides a representative XRPD pattern for Form VLC of Compound (I).

[0021] FIG. 11 provides the IR spectrum of Compound (I) referenced in Example (i).

[0022] FIG. 12 provides the ^-NMR spectrum of Compound (I) referenced in Example (i).

[0023] FIG. 13 provides the ¹³C-NMR spectrum of Compound (I) referenced in Example (i).

[0024] FIG. 14 provides the Mass Spectrum of Compound (I) referenced in Example (i).

[0025] FIG. 15 provides peak-labeled XRPD pattern for Form A of Compound (I).

[0026] FIG. 16 provides peak labeled XRPD pattern for Form B of Compound (I).

5. DETAILED DESCRIPTION

(a) Definitions

[0027] As used herein and unless otherwise specified, the terms "solid form" and related terms refer to a physical form which is not predominantly in a liquid or a gaseous state. As used herein and unless otherwise specified, the term "solid form" and related terms, when used herein to refer to Compound (I), refer to a physical form comprising Compound (I) which is not predominantly in a liquid or a gaseous state. Solid forms may be crystalline, amorphous or mixtures thereof. In particular embodiments, solid forms may be liquid crystals. A "single-component" solid form comprising Compound (I) consists essentially of Compound (I). A "multiple-component" solid form comprising Compound (I) comprises a significant quantity of one or more additional species, such as ions and/or molecules, within the solid form. For example, in particular embodiments, a crystalline multiple-component solid form comprising Compound (I) further comprises one or more species non-covalently bonded at regular positions in the crystal lattice.

[0028] As used herein and unless otherwise specified, the term "crystalline" and related terms used herein, when used to describe a substance, modification, material, component or product, unless otherwise specified, mean that the substance, modification, material, component or product is substantially crystalline as determined by X-ray diffraction. See, e.g., Remington: The Science and Practice of Pharmacy, 21^st edition, Lippincott, Williams and Wilkins, Baltimore, MD (2005); The United States Pharmacopeia, 23^rd edition, 1843- 1844 (1995).

[0029] As used herein and unless otherwise specified, the term "crystal forms" and related terms herein refer to solid forms that are crystalline. Crystal forms include single- component crystal forms and multiple-component crystal forms, and include, but are not limited to, polymorphs, solvates, hydrates, and other molecular complexes, as well as salts, solvates of salts, hydrates of salts, other molecular complexes of salts, and polymorphs thereof. In certain embodiments, a crystal form of a substance may be substantially free of amorphous forms and/or other crystal forms. In certain embodiments, a crystal form of a substance may contain less than about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%), 25%o, 30%), 35%o, 40%>, 45% or 50%> of one or more amorphous forms and/or other crystal forms on a weight basis. In certain embodiments, a crystal form of a substance may be physically and/or chemically pure. In certain embodiments, a crystal form of a substance may be about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% physically and/or chemically pure.

[0030] As used herein and unless otherwise specified, the terms "polymorphs,"

"polymorphic forms" and related terms herein, refer to two or more crystal forms that consist essentially of the same molecule, molecules or ions. Like different crystal forms, different polymorphs may have different physical properties such as, for example, melting

temperatures, heats of fusion, solubilities, dissolution rates and/or vibrational spectra, as a result of the arrangement or conformation of the molecules and/or ions in the crystal lattice. The differences in physical properties may affect pharmaceutical parameters such as storage stability, compressibility and density (important in formulation and product manufacturing), and dissolution rate (an important factor in bioavailability). Differences in stability can result from changes in chemical reactivity {e.g. , differential oxidation, such that a dosage form discolors more rapidly when comprised of one polymorph than when comprised of another polymorph) or mechanical changes {e.g., tablets crumble on storage as a kinetically favored polymorph converts to a thermodynamically more stable polymorph) or both (e.g., tablets of one polymorph are more susceptible to breakdown at high humidity). As a result of solubility/dissolution differences, in the extreme case, some solid-state transitions may result in lack of potency or, at the other extreme, toxicity. In addition, the physical properties may be important in processing (for example, one polymorph might be more likely to form solvates or might be difficult to filter and wash free of impurities, and particle shape and size distribution might be different between polymorphs).

[0031] As used herein and unless otherwise specified, the term "solvate" and "solvated," refer to a crystal form of a substance which contains solvent. The term "hydrate" and "hydrated" refer to a solvate wherein the solvent comprises water. "Polymorphs of solvates" refers to the existence of more than one crystal form for a particular solvate composition. Similarly, "polymorphs of hydrates" refers to the existence of more than one crystal form for a particular hydrate composition. The term "desolvated solvate," as used herein, refers to a crystal form of a substance which may be prepared by removing the solvent from a solvate.

[0032] As used herein and unless otherwise specified, the term "amorphous,"

"amorphous form," and related terms used herein, mean that the substance, component or product in question is not substantially crystalline as determined by X-ray diffraction. In particular, the term "amorphous form" describes a disordered solid form, i.e., a solid form lacking long range crystalline order. In certain embodiments, an amorphous form of a substance may be substantially free of other amorphous forms and/or crystal forms. In other embodiments, an amorphous form of a substance may contain less than about 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% of one or more other amorphous forms and/or crystal forms on a weight basis. In certain embodiments, an amorphous form of a substance may be physically and/or chemically pure. In certain embodiments, an amorphous form of a substance may be about 99%, 98%>, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% physically and/or chemically pure.

[0033] Techniques for characterizing crystal forms and amorphous forms include, but are not limited to, thermal gravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray powder diffractometry (XRPD), single-crystal X-ray diffractometry, vibrational spectroscopy, e.g., infrared (IR) and Raman spectroscopy, solid-state and solution nuclear magnetic resonance (NMR) spectroscopy, optical microscopy, hot stage optical microscopy, scanning electron microscopy (SEM), electron crystallography and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility measurements, dissolution measurements, elemental analysis and Karl Fischer analysis. Characteristic unit cell parameters may be determined using one or more techniques such as, but not limited to, X- ray diffraction and neutron diffraction, including single-crystal diffraction and powder diffraction. Techniques useful for analyzing powder diffraction data include profile refinement, such as Rietveld refinement, which may be used, e.g., to analyze diffraction peaks associated with a single phase in a sample comprising more than one solid phase.

Other methods useful for analyzing powder diffraction data include unit cell indexing, which allows one of skill in the art to determine unit cell parameters from a sample comprising crystalline powder.

[0034] As used herein and unless otherwise specified, the terms "about" and

"approximately," when used in connection with a numeric value or a range of values which is provided to characterize a particular solid form, e.g., a specific temperature or temperature range, such as, for example, that describing a melting, dehydration, desolvation or glass transition temperature; a mass change, such as, for example, a mass change as a function of temperature or humidity; a solvent or water content, in terms of, for example, mass or a percentage; or a peak position, such as, for example, in analysis by IR or Raman spectroscopy or XRPD; indicate that the value or range of values may deviate to an extent deemed reasonable to one of ordinary skill in the art while still describing the particular solid form. For example, in particular embodiments, the terms "about" and "approximately," when used in this context, indicate that the numeric value or range of values may vary within 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the recited value or range of values. As used herein, a tilde {i.e., "~") preceding a numerical value or range of values indicates "about" or "approximately."

[0035] As used herein and unless otherwise specified, a sample comprising a particular crystal form or amorphous form that is "substantially pure," e.g., substantially free of other solid forms and/or of other chemical compounds, or is noted to be "substantially" a crystal form or amorphous form, contains, in particular embodiments, less than about 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or 0.1% percent by weight of one or more other solid forms and/or of other chemical compounds. As used herein and unless otherwise specified, a sample or composition that is "substantially free" of one or more other solid forms and/or other chemical compounds means that the composition contains, in particular embodiments, less than about 25%, 20%>, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or 0.1% percent by weight of one or more other solid forms and/or other chemical compounds. [0036] As used herein, and unless otherwise specified, the terms "treat," "treating" and "treatment" refer to the eradication or amelioration of a disease or disorder, or of one or more symptoms associated with the disease or disorder. In certain embodiments, the terms refer to minimizing the spread or worsening of the disease or disorder resulting from the

administration of one or more prophylactic or therapeutic agents to a subject with such a disease or disorder. In some embodiments, the terms refer to the administration of a compound provided herein, with or without other additional active agent, after the onset of symptoms of the particular disease.

As used herein, and unless otherwise specified, the terms "prevent," "preventing" and "prevention" refer to the prevention of the onset, recurrence or spread of a disease or disorder, or of one or more symptoms thereof. In certain embodiments, the terms refer to the treatment with or administration of a compound provided herein, with or without other additional active compound, prior to the onset of symptoms, particularly to patients at risk of disease or disorders provided herein. The terms encompass the inhibition or reduction of a symptom of the particular disease. Patients with familial history of a disease in particular are candidates for preventive regimens in certain embodiments. In addition, patients who have a history of recurring symptoms are also potential candidates for the prevention. In this regard, the term "prevention" may be interchangeably used with the term "prophylactic treatment."

[0037] As used herein, and unless otherwise specified, the terms "manage," "managing" and "management" refer to preventing or slowing the progression, spread or worsening of a disease or disorder, or of one or more symptoms thereof. Often, the beneficial effects that a subject derives from a prophylactic and/or therapeutic agent do not result in a cure of the disease or disorder. In this regard, the term "managing" encompasses treating a patient who had suffered from the particular disease in an attempt to prevent or minimize the recurrence of the disease.

[0038] Solid forms may exhibit distinct physical characterization data that are unique to a particular solid form, such as the crystal forms described herein. These characterization data may be obtained by various techniques known to those skilled in the art, including for example X-ray powder diffraction, differential scanning calorimetry, thermal gravimetric analysis, and nuclear magnetic resonance spectroscopy. The data provided by these techniques may be used to identify a particular solid form. One skilled in the art can determine whether a solid form is one of the forms described herein by performing one of these characterization techniques and determining whether the resulting data "matches" the reference data provided herein, which is identified as being characteristic of a particular solid form. Characterization data that "matches" those of a reference solid form is understood by those skilled in the art to correspond to the same solid form as the reference solid form. In analyzing whether data "match," a person of ordinary skill in the art understands that particular characterization data points may vary to a reasonable extent while still describing a given solid form, due to, for example, experimental error and routine sample-to-sample analysis.

[0039] In addition to solid forms comprising Compound (I), provided herein are solid forms comprising prodrugs of Compound (I), also provided herein are the methods of making Compound (I) and intermediates leading to Compound (I).

[0040] Solid forms provided herein may also comprise unnatural proportions of atomic isotopes at one or more of the atoms in Compound (I). For example, the compound may be radiolabeled with radioactive isotopes, such as for example deuterium (²H), tritium (³H), iodine-125 (¹²⁵I), sulfur-35 (³⁵S), or carbon-14 (¹⁴C). Radiolabeled compounds are useful as therapeutic agents, e.g., cancer therapeutic agents, research reagents, e.g., binding assay reagents, and diagnostic agents, e.g., in vivo imaging agents. All isotopic variations of Compound (I), whether radioactive or not, are intended to be encompassed within the scope of the embodiments provided herein.

[0041] The solid forms provided herein may be crystalline, amorphous, or an

intermediate form. The crystal forms described herein, therefore, may have varying degrees of crystallinity or lattice order. The solid forms described herein are not limited to any particular degree of crystallinity or lattice order, and may be 0 - 100% crystalline. Methods of determining the degree of crystallinity are known to those of ordinary skill in the, such as those described in Suryanarayanan, R., X-Ray Power Diffractometry, Physical

Characterization of Pharmaceutical Salts, H.G. Brittain, Editor, Mercel Dekkter, Murray Hill, N.J., 1995, pp. 187 - 199, which is incorporated herein by reference in its entirety. In some embodiments, the solid forms described herein are about 0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100 % crystalline,

(b) Solid Forms Comprising Compound (I)

[0042] Provided herein are solid forms comprising the compound of formula (I). H₃CO

OCH

herein, including the methods described in the Examples below, or by techniques known in the art, including heating, cooling, freeze drying, lyophilization, quench cooling the melt, rapid solvent evaporation, slow solvent evaporation, solvent recrystallization, antisolvent addition, slurry recrystallization, crystallization from the melt, desolvation, recrystallization in confined spaces such as, e.g., in nanopores or capillaries, recrystallization on surfaces or templates such as, e.g., on polymers, recrystallization in the presence of additives, such as, e.g., co-crystal counter-molecules, desolvation, dehydration, rapid cooling, slow cooling, exposure to solvent and/or water, drying, including, e.g., vacuum drying, vapor diffusion, sublimation, grinding (including, e.g., cryo-grinding and solvent-drop grinding), microwave- induced precipitation, sonication-induced precipitation, laser-induced precipitation and precipitation from a supercritical fluid. The particle size of the resulting solid forms, which can vary, {e.g., from nanometer dimensions to millimeter dimensions), can be controlled, e.g., by varying crystallization conditions, such as, e.g., the rate of crystallization and/or the crystallization solvent system, or by particle-size reduction techniques, e.g., grinding, milling, micronizing or sonication.

[0044] While not intending to be bound by any particular theory, certain solid forms are characterized by physical properties, e.g., stability, solubility and dissolution rate, appropriate for clinical and therapeutic dosage forms. Moreover, while not wishing to be bound by any particular theory, certain solid forms are characterized by physical properties {e.g. , density, compressibility, hardness, morphology, cleavage, stickiness, solubility, water uptake, electrical properties, thermal behavior, solid-state reactivity, physical stability, and chemical stability) affecting particular processes {e.g. , yield, filtration, washing, drying, milling, mixing, tableting, flowability, dissolution, formulation, and lyophilization) which make certain solid forms suitable for the manufacture of a solid dosage form. Such properties can be determined using particular analytical chemical techniques, including solid-state analytical techniques (e.g., X-ray diffraction, microscopy, spectroscopy and thermal analysis), as described herein and known in the art.

[0045] Certain embodiments herein provide compositions comprising one or more of the solid forms. Certain embodiments provide compositions of one or more solid forms in combination with other active ingredients. Certain embodiments provide methods of using these compositions in the treatment, prevention or management of diseases and disorders including, but not limited to, HCV infection.

[0046] In certain embodiments, provided herein are crystal forms of Compound (I), including but not limited to Form A crystal form, Form B crystal form, Form LC-B crystal form, and Form VLC crystal form, as well as an amorphous form, which are described in more detail below.

[0047] In certain embodiments, the crystal forms are obtained by a method comprising (1) contacting Compound (II) with HC1 and (2) crystallization.

[0048] In some embodiments, crystallization is performed in solvent, water or solvent/water mixtures including, but not limited to, common laboratory organic solvents. In some embodiments, the solvent is a polar or non-polar solvent, or mixture thereof. In some embodiments, the solvent is methanol, ethanol, water, acetone, acetonitrile, 2-butanone, dichloromethane, /?-dioxane, ethyl acetate, isopropanol, methylene chloride, nitromethane, tetrahydrofuran, trifluorotoluene, diethyl ether, tert-butyl methyl ether, n-hexane, c-hexane, dimethoxyethane, and mixtures thereof. In certain embodiments, the solvent is anhydrous,

(i) FORM A CRYSTAL FORM

[0049] Certain embodiments herein provide the Form A crystal form of Compound (I).

[0050] In some embodiments, crystallization of Form A is performed in a solvent system that comprises ethanol, isopropanol, tert-butyl methyl ether, or a mixture thereof.

[0051] In certain embodiments, crystallization methods include, but are not limited to, precipitation, slurry at ambient temperature, slurry at elevated temperature, slurry at sub- ambient temperature, evaporation, slow evaporation, fast evaporation and/or concentration. In certain embodiments, crystallization comprises precipitation with tert-butyl methyl ether.

[0052] In certain embodiments, the Form A crystal form is obtained by a method comprising (1) contacting Compound (II) with HCl in the presence of ethanol and isoproanol to obtain Compound (I); (2) causing precipitation with tert-butyl methyl ether to obtain a slurry; and (3) heating the slurry to about 35-50 °C. In some embodiments, the slurry is heated to about 40 to 45 °C. In some embodiments, the slurry is heated to about 42 °C. In some embodiments, about 2 molar equivalents of HCl relative to Compound (II) are used. In further embodiments, in step (3), the temperature of about 42 °C is maintained for about 1-10, 2-7, or 6 days. In further embodiments, the method further comprises cooling to ambient conditions after step (3). In further embodiments, the method further comprises cooling to about 2 - 8 °C after step (3). In some embodiments, the method further comprises cooling to about 2 - 8 °C after step (3) and maintaining the temperature of about 2 - 8 °C for about 2-48 hours, about 12-36 hours, or about 1 day. In further embodiments, the method comprises isolating the Form A crystal form by vacuum filtration and drying. In further embodiments, the method further comprises seeding.

[0053] A representative XRPD pattern of Form A is provided in FIG. 5. In certain embodiments, Form A of Compound (I) is characterized by XRPD peaks located at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or all of the following approximate positions: 1.54, 3.25, 5.74, 6.61, 8.80, 10.33, 11.44, 11.86, 13.30, 14.50, 15.13, 15.49, 16.51, 17.47, 18.13, 19.42, 20.20, 21.49, 23.05, 24.07, 24.97, 25.87, 27.19, and 28.42 degrees 2Θ, plus or minus 0.10. In certain embodiments, provided herein is a solid form of Compound (I) having an XRPD pattern comprising peaks at approximately 3.25, 5.74, 10.33, 16.51 degrees 2Θ. In further embodiments, the XRPD pattern further comprises peaks at approximately 15.13, 17.47, 23.05, and 24.97 degrees 2Θ. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by an XRPD diffraction pattern which matches the XRPD pattern presented in FIG 5.

[0054] Representative thermal characteristics of the Form A crystal form of Compound (I) are shown in FIG 6 and FIG 7. A representative differential scanning calorimetry (DSC) thermogram, presented in FIG 6, comprises thermal events with maxima at approximately 82 and 216 °C. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by a DSC thermogram comprising one or more thermal events with the following approximate temperature maxima: 82 and 216 °C. In certain embodiments, the temperature maximum at 82 °C corresponds to a loss of solvent, such as, e.g., loss of water, and/or loss of the hydrochloric acid component of the Compound (I). In certain embodiments, the thermal event at 82 °C corresponds to weight loss as detected by TGA. In certain embodiments, the temperature maximum at 216 °C corresponds to decomposition of Compound (I). In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by a DSC thermogram which matches the DSC thermogram presented in FIG. 6.

[0055] A representative thermal gravimetric analysis curve of Form A is provided in FIG 7, which comprises a weight loss of about 8% of the total sample weight upon heating from about 25 - 170 °C. In certain embodiments, the aforementioned weight loss comprises loss of solvent, such as, e.g., a loss of water. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by a TGA

thermogram which matches the TGA thermogram presented in FIG. 7.

[0056] In certain embodiments, provided herein is a solid form comprising Compound (I) that exhibits one or more of the following characteristics as determined by hot stage microscope analysis when heated to about 217 °C: (1) the solid form is birefringent at about 26 °C; (2) the solid form exhibits a slight change in birefringence at about 82 °C; (3) the solid form starts to melt at about 183 °C; (4) the solid form is completely melted at about 207 °C; (5) the solid form becomes slightly brown in color and/or decomposes at about 214 °C.

[0057] In certain embodiments, the chemical profile of a sample of Form A of Compound (I) can be characterized by solution NMR analysis. A representative 1H NMR spectrum of a sample of Form A dissolved in DMSO-d6 is provided in FIG 8.

(ii) FORM B CRYSTAL FORM

[0058] Certain embodiments herein provide the Form B crystal form of Compound (I).

[0059] In some embodiments, crystallization of Form B is performed in a solvent system that comprises ethanol, isopropanol, tert-butyl methyl ether, or a mixture of two or more thereof.

[0060] In certain embodiments, crystallization methods include, but are not limited to, precipitation, slurry at ambient temperature, slurry at elevated temperature, slurry at sub- ambient temperature, evaporation, slow evaporation, fast evaporation and/or concentration. In certain embodiments, crystallization comprises precipitation with tert-butyl methyl ether.

[0061] In certain embodiments, the Form B crystal form is obtained by a method comprising (1) contacting Compound (II) with HCl in the presence of ethanol and isoproanol to obtain Compound (I); (2) causing precipitation with tert-butyl methyl ether to obtain a slurry; (3) heating the slurry to about 35-50 °C; and (4) cooling to ambient temperature. In some embodiments, the slurry is heated to about 40 to 45 °C. In some embodiments, the slurry is heated to about 42 °C. In some embodiments, about 2 molar equivalents of HC1 relative to Compound (II) are used. In further embodiments, in step (3), the temperature of about 42 °C is maintained for about 1-6, 3-5, or 4 days. In further embodiments, the method comprises isolating the Form B crystal form by vacuum filtration and drying. In further embodiments, the method further comprises seeding.

[0062] In certain embodiments, provided herein are methods for synthesizing the Form B crystal form comprising the steps of

(a) contacting the compound of formula (II):

(Π)

with HC1 in the presence of ethanol at 40 - 60 °C to form a mixture;

(b) heating the mixture for 1 - 6 hours at 40 - 60 °C;

(c) concentrating the mixture to dryness to obtain a solid;

(d) heating the solid of step (c) in refluxing methanol for 3 - 8 hours in the

presence of activated carbon;

(e) filtering off the activated carbon to obtain a filtrate and concentrating said filtrate;

(f) heating the concentrated filtrate of step (e) in the presence of acetone for 40

60 °C for 12 - 36 hours and subsequently cooling to room temperature to obtain a suspension;

(g) stirring the suspension of step (f) for 12 - 36 hours; and

(h) filtering the suspension.

[0063] In some embodiments, the compound of formula (I) is contacted with HC1 in the presence of ethanol at about 40, 45, 50, 55, or 60 °C. In some embodiments, the temperature is about 50 °C. In some embodiments, the heating of step (b) is conducted for about 1, 2, 3, 4, 5, or 6 hours. In some embodiments, the heating is conducted for about 5 hours. In some embodiments, the temperature of step (b) is about 40, 45, 50, 55, or 60 °C. In some embodiments, the temperature is about 50 °C. In some embodiments, the heating of step (d) is performed for about 3, 4, 5, 6, 7, or 8 hours. In some embodiments, the heating of step (d) is performed for about 5 hours. In some embodiments, the filtering of step (e) is performed using a 1.0 micron in-line filter. In some embodiments, the heating of step (f) is performed for about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 hours. In some embodiments, the heating of step (f) is performed for about 20 hours. In some embodiments, the temperature of step (f) is about 40, 45, 50, 55, or 60 °C. In some embodiments, the temperature is about 50 °C. In some embodiments, the stirring of step (g) is performed for about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 hours. In some embodiments, the heating of step (g) is performed for about 24 hours.

[0064] A representative XRPD pattern of Form B is provided in FIG. 2. In certain embodiments, Form B of Compound (I) is characterized by XRPD peaks located at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or all of the following approximate positions: 1.54, 3.25, 7.27, 9.67, 11.26, 13.09, 14.53, 16.48, 18.13, 18.73, 20.86, 22.81, 24.01, 26.47, and 28.24 degrees 2Θ, plus or minus 0.10. In certain embodiments, provided herein is a solid form of

Compound (I) having an XRPD pattern comprising peaks at approximately 7.27, 9.67, 11.26, 14.53 degrees 2Θ. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by an XRPD diffraction pattern which matches the XRPD pattern presented in FIG 2.

[0065] Representative thermal characteristics of the Form B crystal form of Compound (I) are shown in FIG 3 and FIG 4. A representative differential scanning calorimetry (DSC) thermogram, presented in FIG 3, comprises thermal events with maxima at approximately 82 and 239 °C. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by a DSC thermogram comprising one or more thermal events with the following approximate temperature maxima: 82 and 239 °C. In certain embodiments, the temperature maximum at 82 °C corresponds to a loss of solvent, such as, e.g., loss of water. In certain embodiments, the thermal event at 82 °C corresponds to weight loss as detected by TGA. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by a DSC thermogram which matches the DSC thermogram presented in FIG. 3.

[0066] A representative thermal gravimetric analysis curve of Form B is provided in FIG 4, which comprises a weight loss of about 4% of the total sample weight upon heating from about 25 - 170 °C. In certain embodiments, the aforementioned weight loss comprises loss of solvent, such as, e.g., a loss of water. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by a TGA

thermogram which matches the TGA thermogram presented in FIG. 4.

(A) Form LC-B Crystal Form

[0067] Certain embodiments herein provide a low crystalline form of the Form B crystal form of Compound (I), which is designated as Low Crystalline Form B or Form LC-B Crystal Form.

[0068] In some embodiments, crystallization of Form LC-B is performed in a solvent system that comprises ethanol, isopropanol, tert-butyl methyl ether, or a mixture of two or more thereof. In another embodiment, the solvent system comprises at least one of ethyl acetate and ethanol or a mixture thereof. In another embodiment, the solvent is ethyl acetate. In another embodiments, the solvent is acetonitrile. In another embodiment, the solvent is a mixture of isopropanol and tert-butyl methyl ether.

[0069] In certain embodiments, crystallization methods including, but are not limited to, precipitation, slurry at ambient temperature, slurry at elevated temperature, slurry at sub- ambient temperature, evaporation, slow evaporation, fast evaporation and/or concentration. In certain embodiments, crystallization comprises precipitation with tert-butyl methyl ether.

[0070] In certain embodiments, the Form LC-B crystal form is obtained by a method comprising (1) contacting Compound (II) with HC1 in the presence of ethanol and isopropanol to obtain Compound (I); (2) causing precipitation with tert-butyl methyl ether to obtain a slurry; and (3) heating the slurry to approximately 35-50 °C. In some embodiments, the slurry is heated to about 40 to 45 °C. In some embodiments, the slurry is heated to about 42 °C. In some embodiments, about 2 molar equivalents of HC1 relative to Compound (II) are used. In certain embodiments, in step (3), the temperature of 42 °C is maintained for about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days. In further embodiments, the method further comprises cooling to ambient conditions after step (3). In further embodiments, the method comprises isolating the Form LC-B crystal form by vacuum filtration and drying. In further embodiments, the method further comprises seeding.

[0071] In certain embodiments, the Form LC-B crystal form is obtained by a method comprising (1) contacting Compound (II) with HC1 in the presence of isopropanol to obtain Compound (I); (2) adding tert-butyl methyl ether; and (3) heating the slurry to approximately 42 °C. In certain embodiments, in step (3), the temperature of 42 °C is maintained for about 1, 2, 3, 4, 5, or 6 days. In some embodiments, about 2 molar equivalents of HC1 relative to Compound (II) are used. In further embodiments, the method further comprises cooling to ambient conditions after step (3). In further embodiments, the method comprises isolating the Form LC-B crystal form by vacuum filtration and drying. In further embodiments, the method further comprises seeding.

[0072] In certain embodiments, the Form LC-B crystal form is obtained by a method comprising (1) contacting Compound (II) with HC1 in the presence of ethyl acetate and ethanol; (2) heating to about 66 °C; (3) cooling to about 35 °C; and (4) cooling to ambient temperature. In some embodiments, in step (2), the temperature of about 66 °C is maintained for about 5 hours. In some embodiments, in step (3), the temperature of about 35 °C is maintained overnight.

[0073] In certain embodiments, the Form LC-B crystal form is obtained by a method comprising contacting Compound (II) with HC1 in the presence of ethyl acetate at room temperature for about 3 days.

[0074] In certain embodiments, the Form LC-B crystal form is obtained by a method comprising contacting Compound (II) with HC1 in the presence of acetonitrile at room temperature for about 3 days.

[0075] A representative XRPD pattern of Form LC-B is provided in FIG. 1. In certain embodiments, Form LC-B of Compound (I) is characterized by XRPD peaks located at one, two, three, four, five, six, or all of the following approximate positions: 7.27, 9.67, 11.26, 14.53, 18.73, 20.86 and 24.01 degrees 2Θ, plus or minus 0.10. In certain embodiments, provided herein is a solid form of Compound (I) having an XRPD pattern comprising peaks at approximately 9.67, 11.26, 18.73, and 20.86 degrees 2Θ. In further embodiments, the XRPD pattern further comprises peaks at about 7.27, 14.53 and 24.01 degrees 2Θ. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by an XRPD diffraction pattern which matches the XRPD pattern presented in FIG 1.

[0076] In certain embodiments, Form LC-B of Compound (I) may be characterized by dynamic vapor sorption or desorption analysis. For example, in one embodiment, a mass loss of about 4% accompanies an initial equilibration at about 5% RH; this is followed by a total mass gain of about 12% when humidity is increased from 5% to 95% RH; and this is followed by a total mass loss of about 13 % when humidity is decreased from about 95% to about 5% RH. In certain embodiments, the resulting material is characterized as Form LC-B of Compound (I), indicating the stability of Form LC-B in the presence of humidity. A representative dynamic vapor sorption/desorption curve for Form B of the HC1 salt of Compound (I) is presented in FIG 9. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by a DVS plot which matches the DVS plot presented in FIG 9.

(iii) FORM VLC CRYSTAL FORM

[0077] Certain embodiments herein provide a solid form comprising compound (I) that is a very low crystalline form, which is designated as Form VLC crystal form of Compound (I).

[0078] In some embodiments, crystallization of Form VLC is performed in a solvent system that comprises ethanol, isopropanol, and tert-butyl methyl ether. In certain embodiments, the solvent is ethyl acetate. In some embodiments, the solvent is a mixture of isopropanol and tert- butyl methyl ether.

[0079] A representative XRPD pattern of Form VLC is provided in FIG. 10. In certain embodiments, Form B of Compound (I) is characterized by XRPD peaks located at one or more of the following approximate positions: 3. degrees 2Θ. In certain embodiments, provided herein is a solid form of Compound (I) having an XRPD pattern comprising peaks at approximately 1.54 and 20.86 degrees 2Θ. In certain embodiments, provided herein is a solid form comprising Compound (I), wherein the solid form is characterized by an XRPD diffraction pattern which matches the XRPD pattern presented in FIG 10.

(iv) AMORPHOUS FORM

[0080] Certain embodiments herein provide solid form comprising compound (I) that is an amorphous form. In some embodiments, the amorphous form is synthesized in a method comprising (1) contacting Compound (II) with HCl in the presence of isopropanol; (2) adding tert-butyl methyl ether; (3) heating to about 42 °C; and (4) cooling to room temperature. In some embodiments, the heating of step 3 is performed for about 4-7 or 6 days. In some embodiments, the amorphous form is synthesized by heating the Form A crystal form at about 80 °C for about 1 day.

(c) Pharmaceutical Compositions

[0081] Certain embodiments provided herein are pharmaceutical compositions comprising the solid forms described herein. In a first embodiment, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients or vehicles, and optionally other therapeutic and/or prophylactic ingredients. Such excipients are known to those skilled in the art.

[0082] Depending on the intended mode of administration, the pharmaceutical compositions may be in the form of solid or semi-solid dosage forms, such as, for example, tablets, suppositories, pills, capsules, powders, suspensions, creams, ointments, lotions or the like, and in some embodiments, in unit dosage form suitable for single administration of a precise dosage. The compositions will include an effective amount of the selected drug in combination with a pharmaceutically acceptable carrier and, in addition, may include other pharmaceutical agents, adjuvants, diluents, buffers, etc.

[0083] The invention includes a pharmaceutical composition comprising a solid form described herein together with one or more pharmaceutically acceptable carriers and optionally other therapeutic and/or prophylactic ingredients.

[0084] For solid compositions, conventional nontoxic solid carriers include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, magnesium carbonate and the like.

[0085] For oral administration, the composition will generally take the form of a tablet, capsule, or suspension. Tablets and capsules are preferred oral administration forms. Tablets and capsules for oral use will generally include one or more commonly used carriers such as lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. When liquid suspensions are used, the active agent may be combined with

emulsifying and suspending agents. If desired, flavoring, coloring and/or sweetening agents may be added as well. Other optional components for incorporation into an oral formulation herein include, but are not limited to, preservatives, suspending agents, thickening agents and the like.

[0086] In some embodiments, provided herein are dosage forms consisting of the solid form alone, i.e., a solid form without any excipients. In some embodiments, provided herein are sterile dosage forms comprising the solid forms described herein.

[0087] Certain embodiments herein provide the use of the solid forms described herein in the manufacture of a medicament. In further embodiments, the medicament is for the treatment of hepatitis C. In some embodiments, the medicament if for the treatment of chronic HCV infection.

(d) Methods of Use

[0088] Certain embodiments herein provide a method of treating HCV infection comprising administering to a subject in need thereof, a therapeutically effective amount of a solid form described herein, optionally in a pharmaceutical composition. In some embodiments, the HCV infection is chronic HCV infection. A pharmaceutically or therapeutically effective amount of the composition will be delivered to the subject. The precise effective amount will vary from subject to subject and will depend upon the species, age, the subject's size and health, the nature and extent of the condition being treated, recommendations of the treating physician, and the therapeutics or combination of therapeutics selected for administration. Thus, the effective amount for a given situation can be determined by routine experimentation. The subject may be administered as many doses as is required to reduce and/or alleviate the signs, symptoms or causes of the disorder in question, or bring about any other desired alteration of a biological system. One of ordinary skill in the art of treating such diseases will be able, without undue experimentation and in reliance upon personal knowledge and the disclosure of this application, to ascertain a therapeutically effective amount of the compounds of this invention for a given disease,

(e) Combination Therapy

[0089] The solid forms and pharmaceutical compositions described herein are useful in treating and preventing HCV infection, including for example chronic HCV infection, alone or when used in combination with other compounds targeting viral or cellular elements or functions involved in the HCV lifecycle. Classes of compounds useful in the invention may include, without limitation, all classes of HCV antivirals. For combination therapies, mechanistic classes of agents that may be useful when combined, including for example, nucleoside and non-nucleoside inhibitors of the HCV polymerase, protease inhibitors, helicase inhibitors, NS4B inhibitors and medicinal agents that functionally inhibit the internal ribosomal entry site (IRES) and other medicaments that inhibit HCV cell attachment or virus entry, HCV RNA translation, HCV RNA transcription, replication or HCV maturation, assembly or virus release. Specific compounds in these classes include, but are not limited to, macrocyclic, heterocyclic and linear HCV protease inhibitors such as telaprevir (VX-950), boceprevir (SCH-503034), narlaprevir (SCH-900518), ITMN-191 (R-7227), TMC-435350 (a.k.a. TMC-435), MK-7009, BI-201335, BI-2061 (ciluprevir), BMS-650032, ACH-1625, ACH-1095 (HCV NS4A protease co-factor inhibitor), VX-500, VX-813, PHX-1766, PHX2054, IDX-136, IDX-316, ABT-450, EP-013420 (and congeners) and VBY-376; the Nucleosidic HCV polymerase (replicase) inhibitors useful in the invention include, but are not limited to, R7128, PSI-7851, IDX-184, IDX-102, R1479, UNX-08189, PSI-6130, PSI- 938, PSI-879 and PSI-7977 and various other nucleoside and nucleotide analogs and HCV inhibitors including (but not limited to) those derived as 2'-C-methyl modified nucleos(t)ides, 4'-aza modified nucleos(t)ides, and 7'-deaza modified nucleos(t)ides. Non-nuclosidic HCV polymerase (replicase) inhibitors useful in the invention, include, but are not limited to, HCV-796, HCV-371, VCH-759, VCH-916, VCH-222, ANA-598, MK-3281, ABT-333, ABT-072, PF-00868554, BI-207127, GS-9190, A-837093, JKT-109, GL-59728 and GL- 60667.

[0090] In addition, solid forms and compositions described herein may be used in combination with cyclophyllin and immunophyllin antagonists (e.g., without limitation, DEBIO compounds, NM-811 as well as cyclosporine and its derivatives), kinase inhibitors, inhibitors of heat shock proteins (e.g., HSP90 and HSP70), other immunomodulatory agents that may include, without limitation, interferons (-alpha, -beta, -omega, -gamma, -lambda or synthetic) such as Intron A™, Roferon-A™, Canferon-A300™, Advaferon™, Infergen™, Humoferon™, Sumiferon MP™, Alfaferone™, IFN-β™, Feron™ and the like; polyethylene glycol derivatized (pegylated) interferon compounds, such as PEG interferon-a-2a

(PegasysTM), PEG interferon-a-2b (PEGIntron™), pegylated IFN-a-conl and the like; long acting formulations and derivatizations of interferon compounds such as the albumin- fused interferon, Albuferon™ , Locteron™ and the like; interferons with various types of controlled delivery systems (e.g. ITCA-638, omega-interferon delivered by the DUROS™ subcutaneous delivery system); compounds that stimulate the synthesis of interferon in cells, such as resiquimod and the like; interleukins; compounds that enhance the development of type 1 helper T cell response, such as SCV-07 and the like; TOLL-like receptor agonists such as CpG-10101 (actilon), isotorabine, ANA773 and the like; thymosin a -1; ANA-245 and ANA-246; histamine dihydrochloride; propagermanium; tetrachlorodecaoxide; ampligen; IMP-321; KRN-7000; antibodies, such as civacir, XTL-6865 and the like and prophylactic and therapeutic vaccines such as InnoVac C, HCV E1E2/MF59 and the like. In addition, any of the above-described methods involving administering an NS5A inhibitor, a Type I interferon receptor agonist (e.g., an IFN-a) and a Type II interferon receptor agonist (e.g., an IFN-γ) can be augmented by administration of an effective amount of a TNF-a antagonist. Exemplary, non-limiting TNF-a antagonists that are suitable for use in such combination therapies include ENBREL™, REMICADETM and HUMIRA™.

[0091] In addition, solid forms and compositions described herein may be used in combination with antiprotozoans and other antivirals thought to be effective in the treatment of HCV infection, and in some embodiments, chronic HCV infection, such as, without limitation, the prodrug nitazoxanide. Nitazoxanide can be used as an agent in combination the compounds disclosed in this invention as well as in combination with other agents useful in treating HCV infection, and in some embodiments, chronic HCV infection, such as peginterferon alfa-2a and ribavarin

(see, for example, Rossignol, JF and Keeffe, EB, Future Microbiol. 3:539-545, 2008). [0092] The solid forms and compositions described herein may also be used with alternative forms of interferons and pegylated interferons, ribavirin or its analogs (e.g., tarabavarin, levoviron), microRNA, small interfering RNA compounds (e.g., SIRPLEX-140- N and the like), nucleotide or nucleoside analogs, immunoglobulins, hepatoprotectants, antiinflammatory agents and other inhibitors of NS5A. Inhibitors of other targets in the HCV lifecycle include NS3 helicase inhibitors; NS4A co-factor inhibitors; antisense

oligonucleotide inhibitors, such as ISIS- 14803, AVI-4065 and the like; vector-encoded short hairpin RNA (shRNA); HCV specific ribozymes such as heptazyme, RPI, 13919 and the like; entry inhibitors such as HepeX-C, HuMax-HepC and the like; alpha glucosidase inhibitors such as celgosivir, UT-231B and the like; KPE-02003002 and BIVN 401 and IMPDH inhibitors. Other illustrative HCV inhibitor compounds include those disclosed in the following publications: U.S. Pat. No. 5,807,876; U.S. Pat. No. 6,498,178; U.S. Pat. No.

6,344,465; U.S. Pat. No. 6,054,472; WO97/40028; WO98/40381; WO00/56331, WO

02/04425; WO 03/007945; WO 03/010141; WO 03/000254; WO 01/32153; WO 00/06529; WO 00/18231; WO 00/10573; WO 00/13708; WO 01/85172; WO 03/037893; WO

03/037894; WO 03/037895; WO 02/100851; WO 02/100846; EP 1256628; WO 99/01582; WO 00/09543; WO02/18369; W098/17679, WO00/056331; WO 98/22496; WO 99/07734; WO 05/073216, WO 05/073195 and WO 08/021927, the entireties of which are incorporated herein by reference.

[0093] Additionally, combinations of, for example, ribavirin and interferon, may be administered as multiple combination therapy with at least one of solid forms or

compositions described herein. Combinable agents are not limited to the aforementioned classes or compounds and contemplates known and new compounds and combinations of biologically active agents (see, Strader, D.B., Wright, T., Thomas, D.L. and Seeff, L.B., AASLD Practice Guidelines. 1-22, 2009 and Manns, M.P., Foster, G.R., Rockstroh, J.K., Zeuzem, S., Zoulim, F. and Houghton, M., Nature Reviews Drug Discovery. 6:991-1000, 2007, Pawlotsky, J-M., Chevaliez, S. and McHutchinson, J.G., Gastroenterology. 132: 179- 1998, 2007, Lindenbach, B.D. and Rice, CM., Nature 436:933-938, 2005, Klebl, B.M., Kurtenbach, A., Salassidis, K., Daub, H. and Herget, T., Antiviral Chemistry &

Chemotherapy. 16:69-90, 2005, Beaulieu, P.L., Current Opinion in Investigational Drugs . 8:614-634, 2007, Kim, S-J., Kim, J-H., Kim, Y-G., Lim, H-S. and Oh, W-J., The Journal of Biological Chemistry. 48:50031-50041, 2004, Okamoto, T., Nishimura, Y., Ichimura, T., Suzuki, K., Miyamura, T., Suzuki, T., Moriishi, K.and Matsuura, Y., The EMBO Journal. 1- 11, 2006, Soriano, V., Peters, M.G. and Zeuzem, S. Clinical Infectious Diseases . 48:313-320, 2009, Huang, Z., Murray, M.G. and Secrist, J.A., Antiviral Research. 71 :351-362, 2006 and Neyts, J., Antiviral Research. 71 :363-371, 2006, each of which is incorporated by reference in their entirety herein). It is intended that combination therapies described herein include any chemically compatible combination of a compound of this inventive group with other compounds of the inventive group or other compounds outside of the inventive group, as long as the combination does not eliminate the anti-viral activity of the compound of this inventive group or the anti-viral activity of the pharmaceutical composition itself.

[0094] Combination therapy can be sequential, that is treatment with one agent first and then a second agent or it can be treatment with both agents at the same time (concurrently). Sequential therapy can include a reasonable time after the completion of the first therapy before beginning the second therapy. Treatment with both agents at the same time can be in the same daily dose or in separate doses. Combination therapy need not be limited to two agents and may include three or more agents. The dosages for both concurrent and sequential combination therapy will depend on absorption, distribution, metabolism and excretion rates of the components of the combination therapy as well as other factors known to one of skill in the art. Dosage values will also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens and schedules may be adjusted over time according to the individual's need and the professional judgment of the person administering or supervising the administration of the combination therapy.

(f) Synthesis of Compound (I) via compounds (II), A-9, A- 10, and 11

[0095] The following abbreviations are used throughout this application:

ACN Acetonitrile

AcOH Acetic acid

aq Aqueous

Bn Benzyl

Boc t-Butoxycarbonyl

Cbz Benzoxylcarbonoyl

DCE Dichloroethane

DCM Dichloromethane

DEAD Diethyl azodicarboxylate

DEPBT 3-(Diethoxy-phosphoryloxy)-3H-benzo[d][l,2,3] triazin-4-one

DIEA (DIPEA) Diisopropylethylamine

DIBAL Diisobutylaluminium hydride

DMA NN-Dimethylacetamide

DME 1 ,2-Dimethoxyethane

DMF NN-Dimethylformamide

DMSO Dimethylsulfoxide

DMTMM 4-(4,6-Dimethoxy- 1 ,3 ,5-triazin-2-yl)-4-methylmorpholinium chloride DPPA Diphenylphosphoryl azide

dppp l,3-Bis(diphenylphosphino)propane

DTT Dithiothreitol

EDCI 1- Ethyl-3-[3-(dimethylamino) propyl]carbodiimide hydrochloride

EDTA Ethylene diamine tetraacetic acid

EC50 Effective concentration to produce 50% of the maximal effect

ESI Electrospray Ionization

Et₃N, TEA Triethylamine

EtOAc, EtAc Ethyl acetate

EtOH Ethanol

g Gram(s)

h or hr Hour(s)

HATU 2- (7- Aza- 1 H-benzotriazole- 1 -yl)- 1 , 1 ,3 ,3 -tetramethyluronium

hexafluorophosphate

HBTU O-Benzotriazol- 1 -y 1-N,N,N ' ,N ' -tetramethyluronium

hexafluorophosphate

HOBt 1 -Hydroxybenzotriazole

IC₅₀ The concentration of an inhibitor that causes a 50 % reduction in a measured activity

LAH Lithium aluminum hydride

LDA Lithium diisopropylamide

LC-MS Liquid Chramatography Mass Spectrometry

mCPBA m-Chloroperoxybenzoic acid

Mel Methyl Iodide

MeOH Methanol

min Minute(s)

mmol Millimole(s)

Moc Methoxylcarbonyl

NMM 4-Methylmorpholine

NMP N-methylpyrrolidinone

PG Protective Group

PTT Phenyl trimethyl tribromide

Py, Pyr Pyridine

rt Room temperature

TEA Triethylamine

Tf Trifluoromethanesulfonate

TFA Trifluoroacetic acid

TFAA Trifluoroacetic anhydride

THF Tetrahydrofuran

TLC Thin Layer Chromatography

TMSOTf Trimethyisiiyl trifluoromethanesulfonate

[0096] Compound (I) may be synthesized from compound (II), the structure of which is depicted, e.g., in Scheme A.

[0097] In some embodiments, provided herein is a method for preparing Compound (I) comprising contacting Compound (II) with HCl. In further embodiments, the method is performed in the presence of ethanol. In further embodiments, Compound (II) is contacted with HCl in the presence of ethanol at a temperature of about 50 °C. In some embodiments, about 2 molar equivalents of HCl are added. In some embodiments, the HCl is added as a 1.25 M solution in ethanol. In further embodiments, Compound (II) is contacted with HCl in the presence of ethanol at a temperature of about 50 °C, and the temperature is maintained at about 50 °C for about three hours. In another embodiment, after maintaining the temperature at about 50 °C for about three hours, the reaction mixture is concentrated to dryness. In some embodiments, dryness is achieved via azeotropic distillation with a suitable solvent, for example, tert-butyl methyl ether. In further embodiments, the dried material is treated with activated carbon in the presence of methanol at reflux. In particular embodiments, the dried material is treated with activated carbon in the presence of refluxing methanol for about 5 hours. In further embodiments, the carbon is filtered, and the filtrate is concentrated and heated to about 50 °C in the presence of acetone. In further embodiments, the filtrate is heated at about 50 °C in the presence of acetone for about 6 hours and then cooled to room temperature and stirred for an additional 24 hours.

[0098] The synthesis of Compound (II) may be accomplished using various methods, including those described in Schemes A, B, and C.

[0100] Scheme A outlines the general synthetic routes leading to key intermediates and the aryl-acetylene-aryl compounds A-10, 11 and II. More specifically, the phenylimidazole intermediate A-2 can be prepared following published procedures (see, e.g., Huang, W., Pei, J., Chen, B., Pei, W., Ye, X., Tetrahedron 52:10131-10136, 1996; Clemens, J. J., Davis, M. D., Lynch, K. R., Macdonald, T. L., Bioorganic & Medicinal Chemistry Letters 15:3568- 3572, 2005; Pinto, D. J. P., Smallheer, J. M., Corte, J. R., Hu, Z., Cavallaro, C. L., Gilligan, P. J., Quan, M. L., PCTInt. Appl, WO 2007070826, 2007; Bachand, C, Belema, M., Deon, D. H., Good, A. C, Goodrich, J., James, C. A., Lavoie, R., Lopez, O. D., Martel, A., Meanwell, N. A., Nguyen, V. N., Romine, J. L., Ruediger, E. H., Snyder, L. B., St. Laurent, D. R., Yang, F., Langley, D. R., Hamann, L. G., U.S. Pat. Appl. Publ. US 2008/0044379, 2008). The coupling of A-2 with a protected acetylene A-3 leads to A-4 via a transformation generally referred to as Sonogashira reaction {Tetrahedron Lett., 50: 4467-4470, 1975). Representative Sonogashira coupling conditions include Pd(PPh₃)₄, Cul, DIEA, DMF, heating; Pd(PPh₃)₂Cl₂, Cul, Pt-Bu₃, piperidine, DMF, heating; or Pd(OAc)₂, Cul, Pt-Bu₃ or P(o-tol)₃, piperidine, DMF, heating etc. A-4 is converted to A-5 on removal of the Y group, typically under a basic condition, such as K₂C0₃ in MeOH or a fluoride source, such as n- Bu₄NF or CsF in THF, when Y is a trimethylsilyl group. Compound A-10 is formed via another Sonogashira reaction between A-9 and A-2. The nitrogen protecting groups (PG) are removed to give 11. II can be prepared by reacting 11 with (5)-N-methoxycarbonyl-valine under peptide coupling condition, using l-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) and hydroxybenzotriazole (HOBt) with ACN as solvent. Other conditions that can be used in promoting this transformation include, but not limited to, HATU/DIEA or

Et₃N/DMF/rt; DCC/THF/rt; PyAOP/DIEA/DMF/rt; and DMTMM/DIEA/DMF or THF/rt etc. The transformation is typically performed in a solvent or mixture of solvents including DMF, THF, ACN, NMP, EtOAc, isopropyl acetate, acetone, 2-methyltetrahydrofuran, particular conditions will vary depending on the nature of the coupling reagents and the scale of reaction.

Scheme A

[0101] In some embodiments, provided herein are methods for producing a compound of formula (II):

II

comprising contacting the compound of formula A-l 1 :

A-l l

or a salt thereof, with (S)-N-methoxycarbonyl-valine in the presence of peptide coupling reagent or reagents. In some embodiments, the peptide couple reagent or reagents is selected from the group consisting of 2~(7~Aza~i H-benzotriazoie- 1. ~y!)~ 1 , 1 ,3,3-tetraTnellvyluromum he afluorophosphate; dicyclohexylcarbodiimide; benzotriazol- 1 -yl- oxytripyrrolidinophosphonium hexafluorophosphate; 4-(4,6-dimethoxy-l ,3,5-triazin-2-yl)-4- methylmorpholinium chloride; 3-(3-dimethylaminopropyl)carbodiimide;

hydroxybenzotriazole; and l-ethyl-3-(3-dimethylaminopropyl) carbodiimide and

hydroxybenzotriazole .

[0102] In some embodiments, provided herein are methods for producing a compound of formula A-l l :

A-l l

comprising contacting the compound of formula A- 10 with a protecting group cleaving agent:

A-10

wherein PG is a protecting group. In some embodiments, PG is Boc and the cleaving agent is an acid. [0103] In some embodiments, provided herein are methods for producing a compound of formula A- 10:

A-10

comprising contacting the compound of formula A-9

A-9

with the compound of formula A-

A-2

in the presence of a coupling reagent or reagents, wherein PG is, individually at each occurrence, a protecting group; and X is Br, I, or OH.

[0104] In some embodiments, provided herein are methods for producing a compound of formula A-10:

A-10

comprising contacting the compound of formula A-8:

A-8 with the compound of formula A-

A-5

[0105] In further embodiments, the coupling reagents include palladium-based catalysts. In some embodiments, the coupling reagents are Sonogashira coupling reagents. In further embodiments, the coupling reagents are selected from the group consisting of Pd(PPh₃)₄, Cul, and diisopropylethylamine; Pd(PPh₃)₂Cl2, Cul, Pt-Bu₃, and piperidine; Pd(OAc)₂, Cul, Pt- Bu₃, piperidine; and Pd(OAc)₂, Cul, P(o-tol)₃, piperidine.

[0106] In some embodiments, provided herein are methods for producing the compound of formula A-9:

A-9

comprising

(a) contacting the compound of formula A-8:

A-8

with ethynyltrimethylsilane in the presence of piperidine, Pt-Bu₃, Cul, and Pd(PPh₃)₂Cl₂ to form an intermediate ; and

(b) contacting the intermediate of step (a) with K₂C0₃ in the presence of methanol.

[0107] In some embodiments, step (a) is performed at a temperature between 60 -80 °C. In some embodiments, the temperature is about 70 °C. [0108] In further embodiments, the protecting group is selected from the group consisting of t-butoxycarbonyl, benzyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, and 2- (trimethylsilyl)ethoxycarbonyl.

[0109] Scheme B describes alternative synthetic routes to compound II. Typical deprotection of A-2, A-5, A-8 and A-9, followed by coupling with (5)-N-methoxycarbonyl- valine under an amide formation condition gives B-1, B-2, B-3 and B-4, respectively.

Sonogashira reaction of A-2 and B-4 or A-8 and B-2, followed by deprotection gives B-6 or B-8. B-6 or B-8, when coupled with (5)-N-methoxycarbonyl-valine affords II. In yet another alternative route, Sonogashira coupling reaction between B-1 and B-4 or B-2 and B-3 directly generates II.

or

Scheme B

[0110] In some embodiments, provided herein are methods for producing a compound of formula B-8:

comprising contacting the compound of formula B-7:

B-7

with a protecting group cleaving agent, wherein PG is a protecting group.

[0111] In some embodiments, provided herein are methods for producing a compound of formula B-7:

B-7

comprising contacting the compound of formula A-8:

A-8

with the compound of formula B-

B-2

in the presence of a coupling reagent or reagents, wherein X is Br, I, or OH and PG is a protecting group.

[0112] In some embodiments, provided herein are methods for producing the compound of formula B-6:

B-6

comprising contacting the compound of formula B-5 :

B-5

with a protecting group cleaving agent, wherein PG is a protecting group.

[0113] Provided herein are methods for producing the compound of formula B-5:

B-5

comprising contacting the compound of formula A-2:

A-2

with the compound of formula B-4:

B-4 in the presence of a coupling reagent or reagents, wherein X is Br, I, or OH and PG is a protecting group.

[0114] In some embodiments, the protecting group is selected from the group consisting of t-butoxycarbonyl, benzyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, and 2- (trimethylsilyl)ethoxycarbonyl.

[0115] In further embodiments, the coupling reagents include palladium-based catalysts. In some embodiments, the coupling reagents are Sonogashira coupling reagents. In further embodiments, the coupling reagents are selected from the group consisting of Pd(PPh₃)₄, Cul, and diisopropylethylamine; Pd(PPh₃)₂Cl2, Cul, Pt-Bu₃, and piperidine; Pd(OAc)₂, Cul, Pt- Bu₃, piperidine; and Pd(OAc)₂, Cul, P(o-tol)₃, piperidine.

[0116] Scheme C depicts alternative approaches to II. Selective Sonogashira reaction of A-9 (Y= H) or B-4 and C-l gives C-2 or C-3, which is transformed to its corresponding pinnacolborane (C-4 or C-5) or boronic acid. The subsequent Suzuki reaction of C-4 or C-5 and C-6 affords A- 10 or B-5, which undergoes a typical deprotection and amide formation sequence to produce compound II. Similarly, compound II can be readily synthesized using -7 derived from C-4 and C-l as the precursor or by directly coupling C-5 and C-l .

Scheme C

[0117] In some embodiments, provided herein are methods for producing the compound of formula C-2:

C-2

comprising compound the compound of formula A-9

A-9

with the compound of formula C-l :

C-l

in the presence of a coupling reagent or reagents, wherein PG is a protecting group and X is Br, I, or OH.

[0118] In some embodiments, provided herein are methods for producing the compound of formula C-3:

C-3

comprising contacting the compound of formula B-4

B-4

with the compound of formula C-l :

C-l

[0119] In further embodiments, the coupling reagents include palladium-based catalysts. In some embodiments, the coupling reagents are Sonogashira coupling reagents. In further embodiments, the coupling reagents are selected from the group consisting of Pd(PPh₃)₄, Cul, and diisopropylethylamine; Pd(PPh₃)₂Cl2, Cul, Pt-Bu₃, and piperidine; Pd(OAc)₂, Cul, Pt- Bu₃, piperidine; and Pd(OAc)₂, Cul, P(o-tol)₃, piperidine.

[0120] Scheme D illustrates alternative approaches to compound II. Sonogashira reaction of D-l and A-5 or B-2, followed by reduction of the nitro group and ring cyclization with A-7 gives A- 10 or B-7, which can be readily converted to compound II.

Scheme D

[0121] In some embodiments, provided herein are methods for producing the compound of formula D-2:

D-2

comprising contacting the compound of formula A-5 :

A-5

with the compound of formula D-l :

D-l

[0122] In some embodiments, provided herein are methods for producing the compound of formula D-3 :

D-3

comprising contacting the compound of formula B-2

with the compound of formula D-l :

D-l

[0123] In further embodiments, the coupling reagents include palladium-based catalysts. In some embodiments, the coupling reagents are Sonogashira coupling reagents. In further embodiments, the coupling reagents are selected from the group consisting of Pd(PPh₃)₄, Cul, and diisopropylethylamine; Pd(PPh₃)₂Cl2, Cul, Pt-Bu₃, and piperidine; Pd(OAc)₂, Cul, Pt- Bu₃, piperidine; and Pd(OAc)₂, Cul, P(o-tol)₃, piperidine.

[0124] The nitrogen protecting groups, i.e., PG, that are utilized in the approaches above, e.g., schemes A-D, include all suitable protecting groups known to those of skilled in the art. In some embodiments, the protecting group is t-butoxycarbonyl, benzyloxycarbonyl, 2,2,2- trichloroethoxycarbonyl, or 2-(trimethylsilyl)ethoxycarbonyl. Other protecting groups and corresponding protecting group cleaving agents are disclosed in Greene et al, Greene's Protective Groups in Organic Synthesis, 4th ed, John Wiley & Sons, New York, 2006, which is incorporated herein by reference in its entirety. A protecting group cleaving agent, as used herein in connection with an amine protecting group, refers to any compound that is capable of removing an amine protecting group to form the corresponding amine or salt thereof. The peptide coupling reactions described above may be performed under any suitable peptide coupling conditions known to those of skill in the art. Such peptide coupling conditions employ peptide coupling reagents that are known to those of ordinary skill in the art.

Exemplary regents include 2-(7-Aza- 1 H-bers zotriazol e- 1 -y l)-l, 1,3,3 -tetrameihy ί uronium hexafluorophosphate; dicyclohexylcarbodiimide; benzotriazol- 1 -yl- oxytripyrrolidinophosphonium hexafluorophosphate; 4-(4,6-dimethoxy-l ,3,5-triazin-2-yl)-4- methylmorpholinium chloride; 3-(3-dimethylaminopropyl)carbodiimide;

hydroxybenzotriazole; or l-ethyl-3-(3-dimethylaminopropyl) carbodiimide and

hydroxybenzotriazole; or derivatives thereof. In some embodiments, a base is employed, such as, but not limited to, diisopropylethylamine and triethylamine. The aryl and alkyne coupling reactions described herein may be performed under any suitable aryl coupling conditions known to those of skill in the art. Exemplary conditions include palladium catalyst-based systems, such as Sonogashira or Suzuki coupling conditions. In some embodiments, the condistions employ Pd(PPh₃)₄, Cul, and diisopropylethylamine;

Pd(PPh₃)₂Cl₂, Cul, Pt-Bu₃, and piperidine; Pd(OAc)₂, Cul, Pt-Bu₃, piperidine; or Pd(OAc)₂, Cul, P(o-tol)₃, piperidine.

The following Examples are presented by way of illustration, not limitation, (i) SYNTHESIS OF COMPOUND (I)

Scheme 1

Example 1 : Synthesis of (S)-tert-butyl 2-(5-(4-ethvnylphenyl)-lH-imidazol-2-yl)pyrrolidine- l-carboxylate(5)

[0126] Step a. Referring to Scheme 1, to a solution of 2-bromo-l-(4- bromophenyl)ethanone (120 g, 0.43 mol) in CH₃CN (300 mL) was added (5)-N-Boc-Pro-OH (97.0 g, 0.45 mol) and Et₃N (130 g, 1.29 mol), the mixture was stirred at room temperature for 2 hrs. The mixture was concentrated under reduced pressure to afford 2. The crude product was used for next step without further purification.

[0127] Step b. To a solution of 2 (159 g, 0.39 mol) in xylene (250 mL) was added NH₄OAc (300 g, 3.90 mol), the mixture was stirred at 140 °C for overnight. The mixture was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (petroleum ether/EtOAc = 10/1 (v/v)) to afford 3 (105 g, 70% yield) as a white solid: 1H NMR (500 MHz, CDC1₃) δ 1.48 (s, 9H), 1.96 (m, 1H), 2.16 (m, 2H), 3.01 (m, 1H), 3.42 (m, 2H), 4.96 (d, 1H, J= 5.5Hz), 7.22 (s, 1H), 7.46-7.55 (m, 4H) ppm; LC-MS (ESI): m/z 392.1 (M+H)⁺.

[0128] Step c. To a solution of 3 (10.0 g, 25.5 mmol) in anhydrous THF (100 mL) were added PPh₃ (1.34 g, 5.11 mmol), Pd (PPh₃)₂Cl₂ (1.79 g, 2.56 mmol), Cul (0.24 g, 1.28 mmol), DIPEA (7.75 g, 76.8 mmol), and trimethylsilylacetylene (5.02 g, 51.2 mmol), the mixture was refluxed under argon for overnight. The organic solvent was removed under reduced pressure and residue was treated with water, extracted with EtOAc (2 x 100 mL), the combined organic phase was dried, filtered, and concentrated in vacuo to give a residue, which was purified by silica gel column chromatography (petroleum ether/EtOAc = 3/1 (v/v)) to afford 4a (5.80 g, 55% yield) as a yellow solid: 1H NMR (500 MHz, CDC1₃) δ 0.21 (s, 9H), 1.49 (s, 9H), 1.97 (m, 1H), 2.16 (m, 2H), 2.40 (brs, 1H), 3.41 (m, 2H), 4.98 (d, 1H, J = 7.0Hz), 6.78 (s, 1H), 7.61 - 8.01 (m, 4H) ppm; LC-MS (ESI): m/z 410.3 (M+H)⁺.

Alternatively, 2-methylbut-3-yn-2-ol was used as the masked source of acetylene in the place of trimethylsilylacetylene. Dichlorobis(triphenylphosphine)palladium (3.30 g, 4.71 mmol) and copper iodide (0.90 g, 4.71 mmol) were added to a stirred suspension of 2-methylbut-3- yn-2-ol (22.99 ml, 235 mmol) and 3 (46.16 g, 118 mmol) in TEA (120 mL ) and heated at 90 °C for 5 hrs. The reaction was allowed to cool and filtered through a pad of charcoal and Si0₂, which was washed with EtOAc. The filtrate was concentrated in vacuo to give dark foam. The crude residue was then purified by silica gel column chromatography

(DCM/EtOAc = 1/1 (v/v)) to give 4b as an off-white solid (35.95 g, 77% yield).

[0129] Step d. To a solution of 4a (5.80 g, 14.1 mmol) in THF (100 mL) and MeOH (100 mL) was added K₂C0₃ (5.85 g, 42.4 mmol), the mixture was stirred at room temperature for 3 hrs. The solvent was removed under reduced pressure and the residue was purified by silica gel column chromatography (DCM / MeOH = 40: 1 (v/v)) to afford 6 (3.80g, 80% yield) as a yellow solid: 1H NMR (500 MHz, CDC1₃) δ 1.49 (s, 9H), 1.97 (m, 1H), 2.15 (m, 2H), 3.01 (brs, 1H), 3.40 (m, 2H), 4.96 (d, 1H, J= 5.0Hz), 7.24 (s, 1H), 7.47 - 7.52 (m, 4H) ppm.

Scheme 2

Example 2: Synthesis of (S)-tert-butyl 2-(6-iodo-lH-benzo[d]imidazol-2-yl)pyrrolidine-l- carboxylate (8a)

[0130] Step a. Referring to Scheme 2, to a stirred solution of 2-nitroaniline (18.8 g, 137 mmol) and NaOAc (12.7 g, 155 mmol) in AcOH (70 mL) was added ICI (25.0 g, 155 mmol) in AcOH (40 mL) slowly over 30 min. The mixture was heated at 50 °C for 30 min, and stirred at room temperature for additional 30 min. The reaction mixture was poured slowly into water (150 mL) while vigorous stirring and the stirring was continued for 17 hrs. The resulting precipitate was collected by filtration, washed with water (100 mL), dried under vacuum to give 2-nitro-4-iodoaniline (35 g, 95%> yield) as a red powder: LC-MS (ESI): m/z 265 (M + H)⁺. [0131] Step b. To a stirred solution of SnCl₂ (78.0 g, 346 mmol) in concentrated HC1 (150 mL) was added 2-nitro-4-iodoaniline (25.4 g, 92.0 mmol) in three potions over 30 min at room temperature. The reaction mixture was heated at 70 °C for 1 h and then cooled. The mixture was treated with water (150 mL) and stirred for 2 hrs. The precipitate was collected by filtration and dried under vacuum to afford 4-iodobenzene-l,2-diamine (6a) (17 g, 81% yield) as a grey solid. LC-MS (ESI): m/z 235 (M + H)⁺.

[0132] Step c. A mixture of 4-iodobenzene-l ,2-diamine (6a) (1.05 g, 4.40 mmol), (S)- tert-butyl 2-formylpyrrolidine-l-carboxylate (7a) (1.01 g, 4.40 mmol) and iodine (0.11 g, 0.44 mmol) in AcOH (5 mL) was stirred at room temperature in open air overnight, then neutralized with aqueous NaHC0₃, extracted with EtOAc (3 x 100 mL). The organic layer was washed with brine, dried over anhydrous Na₂S0₄, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (Petroleum ether/EtOAc = 4/1 (v/v)) to afford (S)-tert-butyl 2-(6-iodo-lH-benzo[d]imidazol-2-yl)pyrrolidine-l-carboxylate (8a) (500 mg, 30% yield) as a yellow solid. LC-MS (ESI): m/z 414 (M + H)⁺.

Scheme 3

Example 3 : Alternative Synthesis of (S)-tert-butyl 2-(6-bromo-lH-benzord1imidazol-2- vDpyrrolidine- 1 -carboxylate(8b)

[0133] Step a. Referring to Scheme 3, to a solution of (S)-N-Boc-Pro-OH (7b) (29 g, 135 mmol) and DIPEA (29 g, 225 mmol) in THF (500 mL) was added HATU (51 g, 135 mmol) at rt. After stirring for 10 min, 4-bromobenzene-l,2-diamine (6b) (25 g, 135 mmol) was added and the resulting solution was stirred at rt for another several hrs until the completion of the reaction. Subsequently, the reaction mixture was concentrated and the residue was diluted with EtOAc (500 mL). The resulting mixture was washed with water for several times (100 mL x 3) and dried with anhydrous Na₂S0₄. The solvent was removed and the residue was dried in vacuo to give a mixture of crude compounds 6c and 6c', which were used for the next step without further purification. LC-MS (ESI): m/z 384.1 (M+H)⁺. [0134] Step b. A mixture of crude compounds 6c and 6c' obtained from the reaction above in AcOH (1000 mL) was stirred at 40 °C for 12 h. Subsequently, the reaction mixture was carefully neutralized by adding saturated aqueous sodium hydroxide solution to adjust the pH to 8. The resulting mixture was extracted with EtOAc for several times (250 mL x 3) The extracts were combined, washed with water, and dried with anhydrous Na₂S04. The solvent was removed and the residue was purified by silica gel chromatography (Petroleum ether/EtOAc = 4/1 (v/v)) to give compound 8b (35 g, 71% yield) as a yellow solid. 1H NMR (500 MHz, CDCI₃) δ 10.82 (br s, 1H), 7.28-7.72 (m, 3H), 5.09 (d, 1H, J= 6.0 Hz), 3.44 (m, 2H), 2.97 (m, 1H), 2.21 (m, 2H), 2.01 (m, 1H), 1.51 (m, 9H) ppm; LC-MS (ESI): m/z 366.1 (M+H)⁺.

Example 4. CSVfert-butyl 2-(5-(4-((2-((_t^-l-(tert-butoxycarbonvnpyrrolidin-2-vn-lH- benzo[dlimidazol-6-yl)ethynyl)phenyl)-lH-imidazol-2-yl)pyrrolidine-l-carboxylate(10)

[0135] A 22-L round bottom flask in a heating mantle was charged with DMF (6.9 L) and brought to reflux while being simultaneously purged with nitrogen for 1 hr. The mixture was then allowed to cool to approximately 70 °C. Compounds 5 (730 g 2.16 mol), 8b (690 g, 1.88 mol), palladium acetate (21 g, 0.09 mol), copper iodide (3.6 g, 0.02 mol) and tri-o- toylphosphine (60 g, 0.19 mol) were then charged to a separate 22-L flask (also in a heating mantle) and the flask was rendered inert with nitrogen. The warm DMF was transferred under inert conditions to the flask containing the solids. The mixture was then agitated and piperidine (1.1 L, 11.1 mol) was added. The reaction was continued at 80 °C and monitored by HPLC. After 1 hr no starting material 5 was detected. The mixture was allowed to cool to ambient temperature and subsequently poured into water with vigorous agitation. The suspension was then filtered to collect the crude product as a yellow solid, which was then dried in a vacuum oven at 55 °C overnight. The crude product (1.4 kg) was then purified via silica gel column chromatography in three batches following procedures detailed below. The crude solid was taken in methanol/methylene chloride (4 L, 1/3 ratio (v/v)) and agitated until dissolved. Silica gel (1.5 kg) was then added and the slurry mixed and concentrated to dryness. After equilibration of the column with 1% methanol/methylene chloride, the yellow/orange power was loaded into a Sample Injection Module (SIM, 2 x 2 L) and then (after equilibration also with 1% MeOH:DCM) loaded onto the Biotage column. The SIM and column were then eluted with 1 x 20 L 1% MeOH/DCM then 1 x 20 L 2% MeOH/DCM and finally 2 x 20 L 3% MeOH/DCM. Three columns combined gave 540 g product 10 (46% yield). 1H NMR was used to confirm the identity. Analysis by HPLC indicated the product had a purity of 99%. 1H NMR (500 MHz, CDC1₃) δ 10.76 (br s, 1H), 10.54 (br s, 1H), 7.25- 7.92 (m, 8H), 5.11 (m, 1H), 4.97 (m, 1H), 3.41 (m, 4H), 3.04 (m, 2H), 2.16-2.20 (m, 4H), 1.97-2.02 (m, 2H), 1.51 (sm 9H), 1.50 (s, 9H) ppm; LC-MS (ESI): m/z 623.3 (M+H)⁺.

Example 5. 2-((6 -pyrrolidin-2-yl)-6-((4-(2-((6 -pyrrolidin-2-yl)- lH-imidazol-5- yl)phenyl)ethvnyl)-lH-benzord1imidazole hydrochloride (11)

[0136] Compound 10 (519 g, 0.83 mol) was taken in 2-propanol (5.2 L) and cooled to 0-5 °C (ice-bath). 12 N HC1 (0.70 L, 8.3 mol) was slowly added to the suspension keeping the temperature <15 °C. Upon complete addition the resulting clear orange solution was then warmed to 60 °C and continued with agitation for 2 hrs. After 2 hrs an in-process check of the yellow suspension showed reaction completion. The mixture was then cooled to room temperature and then filtered, washing with 1.5 L MeOH/IPA (1/1 (v/v)). The product was collected and dried in vacuo to give 631 g. The crude product was then transferred to a 12-L round bottom flask. Methanol (2.5 L) was then added and the mixture warmed to 50 °C and agitated at that temperature for 5 hrs. The mixture was then allowed to cool to room temperature and stirred for an additional 14 hrs (overnight). The suspension was then filtered and the collected product was dried in vacuo to give 389 g of 11 (94% yield). Purity was >99% by HPLC. LC-MS (ESI): m/z 212.1 (M+2H)²⁺ 1H NMR (500 MHz, CD₃OD) δ 8.13 (s, 1H), 7.96 (s, 1H), 7.91 (d, 2H, J= 8.0 Hz), 7.79 (d, 1H, J= 8.5 Hz), 7.74 (d, 2H, J= 8.5 Hz), 7.67 (d, 1H, J= 8.3 Hz), 3.61-3.64 (m, 4H), 2.74 (m, 2H), 2.25-2.59 (m, 6H) ppm; LC-MS (ESI): m/z 423.2 (M+H)⁺.

Example 6. Methyl Ν-Γ(2£)-1 -\(2S)-2-{5-\4-(2- (2-r(2S)- l -r(2S)-2- r(methoxycarbonyl)amino1 -3 -methylbutanoyl1pyrrolidin-2-vH - 1H- 1 ,3 -benzodiazol-6- yl| ethvnvDphenvH- lH-imidazol-2-yllpyrrolidin-l -νΠ-3-methyl- 1 -oxobutan-2-vH carbamate

[0137] A reaction flask, equipped with a mechanical agitator, a thermocouple and nitrogen inlet was sequentially charged with acetonitrile (5.8 L), 1-hydroxybenotraizole (232 g, 1.72 mol), N-(methoxycarbonyl)-L-valine (291 g, 1.65 mol) and l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI, 328 g, 1.72 mol). The mixture was agitated at 0-5 °C (ice-bath) for 30 min and then charged with 11. While maintaining a temperature of <10 °C, diisopropylethylamine (542 mL, 0.742 mol) was added. The mixture was then warmed to 15-20 °C over 2 hrs and agitated for an additional 17 hrs. An in-process check by HPLC showed complete conversion to the desired product (no starting material was detected), at which point the mixture was washed once with 2 L of 13 wt % aqueous NaCl, twice with 2 L 0.5 N NaOH containing 13 wt % aqueous NaCl and once with 2 L of 13 wt % aqueous NaCl. An in-process HPLC indicated residual HOBt remained in the organic fraction (~5%) and therefore an additional wash was performed using 2 L 0.5 N NaOH containing 13 wt % aqueous NaCl. An in-process HPLC indicated residual HOBt was now <1% in the organic fraction. The organic solution was then concentrated which also served to azeotropically remove any residual water. 1H NMR (500 MHz, CD₃OD) δ 7.33- 7.75 (m, 8H), 5.27 (m 1H), 5.16 (m, 1H), 4.26 (m, 2H), 3.99-4.06 (m, 2H), 3.88-3.94 (m, 3H), 3.71 (s, 6H), 2.06-2.43 (, 11H), 0.89-0.99 (m, 12H) ppm; LC-MS (ESI): m/z 737.4 (M+H)⁺.

Example 7. Methyl Ν-Γ(2£)-1 -Γ(2£)-2-{5-Γ4-(2- (2-r(2SVl-r(2SV2- [(methoxycarbonyl)aminol -3 -methylbutanoyllpyrrolidin-2-yll - 1 H- 1 ,3 -benzodiazol-6- yl| ethynyDphenyll - 1 H-imidazol-2-yl| pyrrolidin- 1 -yl] -3 -methyl- 1 -oxobutan-2-yl] carbamate dihydrochloride (I)

[0138] Taking the product from Example 6, a solvent exchange into ethanol was next performed to a final target volume of 3.9 L. While maintaining a temperature of 50 °C, 1.4 L of 1.25 M HCl in ethanol was charged to the solution containing II. The mixture was agitated at 50 °C for 3 hrs and then concentrated. MTBE (~2 x 2 L) was added as an additive for azeotropic distillation to dryness. The resulting solid was then dissolved in methanol (5 L) and treated with 30 g activated carbon at reflux with agitation for 5 hrs. The solution was cooled to room temperature and filtered through a 1.0 micron in-line filter. The filtrate was concentrated to a target volume of 2 L via vacuum distillation. While maintaining a temperature of 50 °C and with agitation, acetone (2 L) was added. Upon complete addition, the mixture was agitated at 50 °C for 3 hrs then cooled to room temperature and allowed to stir for an additional 17 hrs. Due to a lower than expected amount of product, the mixture was warmed back to 50 °C and an additional 1 L of acetone was added to the mixture. The mixture was the allowed to stir at 50 °C for 3 hrs and then cooled to room temperature.

Stirring for an additional 24 hrs led to a milky white suspension which was filtered to collect the product. The filter cake was washed with fert-butylmethyl ether (~1 L) and then transferred to drying trays. The product was dried in the vacuum oven at 40 °C for 18 hrs to give 350 g (55% yield) of the final product. Analysis confirmed the identity and purity was found to be 98.9% by HPLC peak area percent.

Crystalline Form B was obtained, as determined by XRPD, DSC analyses.

[0139] FT-IR: An IR spectrum was obtained using a Bruker Alpha T spectrometer. A potassium bromide (KBr) pellet was formed by mixing 2.14 mg I with 120.0 mg KBr. A spectrum was obtained from 16 scans with a resolution of 8 cm^"1. The IR spectrum is presented in FIG. 11.

[0140] 1H-NMR: The 1H-NMR spectrum was obtained using a Bruker 400 MHz NMR System. Compound I was dissolved in deuterated methanol. The 1H-NMR spectrum (Figure 1) and the chemical shift assignments shown in the table below is consistent with the structure of I. The 1H-NMR is presented in FIG. 12.

[0141] C-NMR: The ^1JC-NMR spectrum (Figure 13) was obtained using a Bruker 400 MHz NMR. I was dissolved in deuterated methanol. The ¹³C-NMR chemical shifts are consistent with the structure of I.

Figure 13: ¹³C-NMR Spectrum of I.

Peak picking of characteristic signals

[0142] Mass Spectrometry

The mass spectrum of compound (I) is provided in FIG. 14.

Scheme 4

Example 8. (S)-tert-butyl 2-(6-ethynyl-lH-benzo[(i imidazol-2-yl)pyrrolidine-l-carboxylate £9bJ

[0143] Step a. Referring to Scheme 4, a mixture of 8b (21.9 g, 0.060 mol),

ethynyltrimethylsilane (23.6 g, 0.24 mol), piperidine (6.1 g, 0.072 mol), Pt-Bu₃ (7.3 g, 0.036 mol), Cul (1.2 g, 0.006 mol) and Pd(PPh₃)₂Cl₂ (4.2 g, 0.006 mol) in DMF (300 mL) was stirred at 70°C overnight under an atmosphere of N₂. The reaction mixture was concentrated and the residue was diluted with dichloromethane (250 mL). Subsequently, the mixture was washed with water (50 mL x 2) and dried with anhydrous Na₂S0₄. The solvent was removed and the residue was purified by column chromatography (EtO Ac/petroleum ether = 1/10 to 1/2 (v/v)) to give 9a (20 g, 87% yield). LC-MS (ESI): m/z 384.2. [0144] Step b. A mixture of 9a (15 g, 0.039 mol) and K₂C0₃ (6.5 g, 0.047 mol) in methanol (200 mL) was stirred at rt for lh. The reaction mixture was filtered through Celite^®545 and the filtered cake was washed with EtOAc (50 mL x 5). The filtrate was concentrated and the residue was diluted with EtOAc (250 mL). Subsequently, the mixture was washed with water (50 mL x 2) and dried with anhydrous Na₂S0₄. The solvent was removed and the residue was purified by column chromatography (EtO Ac/petroleum ether = 1/10 to 1/1 (v/v)) to give 9b (10 g, 82% yield). LC-MS (ESI): m/z 312.2.

(ii) SYNTHESIS OF FORM A CRYSTAL FORM

[0145] Compound II (408.6 mg or 0.5545 mmol) was dissolved in isopropanol (14 mL) with sonication, yielding a clear solution. Hydrochloric acid (1.25 M in ethanol) was added in a 1 :2 API: acid molar ratio with stirring, also resulting in a clear solution. Aliquots of tert- butyl methyl ether (6 x 5 mL) were added with stirring, causing precipitation on contact and resulting in a white opaque solution with white solid in suspension. The slurry was allowed to stir on a hot plate at approximately 42 °C for a total of 6 days. A cloudy solution with a band of white and yellow solid on the walls at the top of the solution was observed. The solid was scraped down into the solution, and the slurry was allowed to continue stirring at approximately 42 °C for approximately 2 hours. A band of white solid at the top edge of the solution was observed at this time. The hot plate was then turned off, allowing the mixture to slowly cool to ambient temperature. No change was observed after cooling and standing at ambient conditions for approximately 2 hours. The sample was placed in the refrigerator (2 - 8 °C) for 1 day, resulting in an opaque solution with white and yellow solid on the walls and in suspension. The solid was collected by vacuum filtration, air dried under reduced pressure for approximately 20 minutes and under nitrogen gas for approximately 2 minutes, and broken up with a spatula while drying.

(iii) SYNTHESIS OF FORM B CRYSTAL FORM

[0146] To a solution of the dihydrochloride salt of compound II, aliquots of tert-bvXy\ methyl ether (5 x 5 mL) were added slowly with stirring, resulting in a mixture of yellow goo on the bottom of the flask and off-white solid in suspension. The slurry was placed on a hot plate/ stirrer at approximately 42 °C and a clear solution with yellow goo was observed after stirring for a few minutes. The slurry was left to stir for 4 days at those conditions, resulting in a white opaque solution with off-white solid caked onto the stir bar. The hot plate was then turned off to allow the mixture to slowly cool to ambient temperature. After cooling to ambient and standing for approximately 2 hours, a thick white opaque solution with solid in suspension was observed. The solid was collected by vacuum filtration, air dried under reduced pressure approximately 3 minutes, and additionally dried under reduced pressure and nitrogen for approximately 6 minutes, resulting in dry off-white solid, which was later determined to be crystalline pattern B by XRPD.

(iv) SYNTHESIS OF FORM LC-B CRYSTAL FORM

Preparation 1

[0147] Compound II (29.9 mg or 0.0406 mmol) was dissolved in isopropanol (1 mL) with sonication, yielding a clear solution. Hydrochloric acid (1.25 M in ethanol) was added in a molar ratio 1/2 of API/ acid with stirring, also resulting in a clear solution. 7¾rt-butyl methyl ether (1 mL) was added with stirring, causing precipitation on contact and resulting in a cloudy solution with white solid present. The slurry was allowed to stir on a hot plate at approximately 44 - 45 °C for 1 day. A cloudy solution with white solid on the walls was observed. The hot plate was then turned off, allowing the mixture to slowly cool to ambient temperature. A cloudy solution with white solid on the walls and in suspension was observed after cooling and standing at ambient conditions for a few hours. The solid was collected by vacuum filtration and air dried under reduced pressure and nitrogen gas.

Preparation 2

[0148] Compound II (405.5 mg or 0.5503 mmol) was dissolved in ethanol: isopropanol 11 :4 (10 mL) with sonication, yielding a clear yellow solution. Hydrochloric acid (1.25 M in ethanol) was added in a 1/2 molar ratio API/ acid with stirring, also resulting in a clear light yellow solution (slight color change). Aliquots of tert-butyl methyl ether (4 x 5 mL) were added with stirring, causing precipitation on contact and resulting in an opaque solution with off- white solid in suspension. The slurry was allowed to stir on a hot plate at approximately 42 °C for 1 day. A cloudy solution with off-white and yellow solid on the walls was observed. The slurry was removed from the hot plate, and a sub-sample was quickly collected. The bulk slurry was returned to the hot plate, and the sub-sample was analyzed by XRPD and found to be low crystalline pattern B.

(v) INSTRUMENTAL TECHNIQUES

(A) Differential Scanning Calorimetry (DSC)

[0149] DSC was performed using a TA Instruments 2920 differential scanning calorimeter. Temperature calibration was performed using NIST traceable indium metal. The sample was placed into an aluminum DSC pan, and the weight was accurately recorded. The pan was covered with a lid, and the lid was crimped. A weighed, crimped aluminum pan was placed on the reference side of the cell. The sample cell was equilibrated at -30 °C and heated under a nitrogen purge at a rate of 10 °C/minute, up to a final temperature of 250 °C.

Reported temperatures are at the transition maxima.

[0150] FIG. 3 provides a representative DSC thermogram for Form B of Compound (I).

[0151] FIG. 6 provides a representative DSC thermogram for Form A of Compound (I).

(B) Dynamic Vapor Sorption/ Desorption (DVS)

[0152] Dynamic vapor sorption/desorption (DVS) data were collected on a VTI SGA-100 Vapor Sorption Analyzer. NaCl and PVP were used as calibration standards. Samples were not dried prior to analysis. Adsorption and desorption data were collected over a range from 5 to 95% RH at 10% RH increments under a nitrogen purge. The equilibrium criterion used for analysis was less than 0.0100% weight change in 5 minutes with a maximum equilibration time of 3 hours. Data were not corrected for the initial moisture content of the samples.

[0153] FIG. 9 provides a representative dynamic vapor sorption/desorption curve for Form LC-B of Compound (I).

(C) Hot Stage Microscopy

[0154] Hot stage microscopy was performed using a Linkam hot stage (model FTIR 600) mounted on a Leica DM LP microscope equipped with a SPOT Insight™ color digital camera. Temperature calibrations were performed using USP melting point standards.

Samples were placed on a cover glass, and a second cover glass was placed on top of the sample. As the stage was heated, each sample was visually observed using a 20x 0.40 N. A. long working distance objective with crossed polarizers and a first order red compensator. Images were captured using SPOT software (v. 4.5.9).

[0155] A sample of Form A crystal form was analyzed by hot stage microscopy. The table below summarizes the observations obtained when the sample was heated to about 217 °C and subsequently cooled to 27.5 °C.

Temperature °C Observation

183.2 start of melt

192.2 melt continuing, no discoloration/ decomposition

206.8 melt complete, no discoloration/ decomposition

214.3 slight light brown color of "puddles"

217.2 heat removed, no change

27.8 no recrystallization

(D) Thermogravimetry (TGA)

[0156] TGA analyses were performed using a TA Instruments 2950 thermogravimetric analyzer. Temperature calibration was performed using nickel and Alumel™. Each sample was placed in an aluminum pan and inserted into the TGA furnace. The furnace was heated under nitrogen at a rate of 10 °C/minute to a final temperature of 350 °C.

[0157] FIG 4. provides a representative TGA thermogram of Form B of Compound (I).

[0158] FIG. 7 provides a representative TGA thermogram of Form A of Compound (I).

(E) X-Ray Powder Diffraction (XRPD)

Inel XRG-3000 Diffractometer

[0159] XRPD patterns were collected using an Inel XRG-3000 diffractometer equipped with a curved position sensitive detector with a 2Θ range of 120°. An incident beam of Cu Ka radiation (40 kV, 30 mA) was used to collect data in real time at a resolution of 0.03° 2Θ. Prior to the analysis, a silicon standard (NIST SRM 640c) was analyzed to verify the Si 111 peak position. Samples were prepared for analysis by packing them into thin-walled glass capillaries. Each capillary was mounted onto a goniometer head and rotated during data acquisition. In general, the monochromator slit was set at 5 mm by 160 μιη, and the samples were analyzed for 5 minutes.

Bruker D-8 Discover Diffractometer

[0160] XRPD patterns were collected using a Bruker D-8 Discover diffractometer and Bruker's General Detector System (GADDS, v. 4.1.20). An incident microbeam of Cu Ka radiation was produced using a fine-focus tube (40 kV, 40 mA), a Gobel mirror, and a 0.5 mm double-pinhole collimator. Prior to the analysis, a silicon standard (NIST SRM 640c) was analyzed to verify the Si 111 peak position. The sample was packed between 3 μιη thick films to form a portable, disc-shaped specimen. The prepared specimen was loaded in a holder secured to a translation stage. A video camera and laser were used to position the area of interest to intersect the incident beam in transmission geometry. The incident beam was scanned and rastered to optimize orientation statistics. A beam-stop was used to minimize air scatter from the incident beam. Diffraction patterns were collected using a Hi-Star area detector located 15 cm from the sample and processed using GADDS. The intensity in the GADDS image of the diffraction pattern was integrated using a step size of 0.04° 2Θ. The integrated patterns display diffraction intensity as a function of 2Θ.

PANalvtical EXPERT Pro MPD Diffractometer

[0161] The XRPD patterns were collected using a PANalytical X'Pert Pro diffractometer. An incident beam of Cu Ka radiation was produced using an Optix long, fine-focus source. An elliptically graded multilayer mirror was used to focus the Cu Ka X-rays of the source through the specimen and onto the detector. Data were collected and analyzed using X'Pert Pro Data Collector software (v. 2.2b). Prior to the analysis, a silicon specimen (NIST SRM 640c) was analyzed to verify the Si 111 peak position. The specimen was sandwiched between 3 um thick films, analyzed in transmission geometry, and rotated to optimize orientation statistics. A beam-stop was used to minimize the background generated by air scattering. Soller slits were used for the incident and diffracted beams to minimize axial divergence. Diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) located 240 mm from the specimen.

Shimadzu XRD-6000 Diffractometer

[0162] XRPD patterns were collected using a Shimadzu XRD-6000 X-ray powder diffractometer. An incident beam of Cu Ka radiation was produced using a long, fine-focus X-ray tube (40 kV, 40 mA) and a curved graphite monochromator. The divergence and scattering slits were set at 1°, and the receiving slit was set at 0.15 mm. Diffracted radiation was detected by a Nal scintillation detector. Data were collected and analyzed using XRD- 6100/7000 software (v. 5.0). Prior to the analysis, a silicon standard (NIST SRM 640c) was analyzed to verify the Si 1 11 peak position. Samples were prepared for analysis by placing them in an aluminum holder with a silicon zero-background insert. Patterns were typically collected using a Θ-2Θ continuous scan at 3 °/min. (0.4 sec/0.02° step) from 2.5 to 40° 2Θ.

[0163] FIG. 1 provides a representative XRPD pattern of Form LC-B of Compound (I).

[0164] FIG. 2 provides a representative XRPD pattern of Form B of Compound (I). [0165] FIG. 5 provides a representative XRPD pattern of Form A of Compound (I).

[0166] FIG. 10 provides a representative XRPD pattern for Form VLC of Compound (I),

(vi) SOLUBILITY PROPERTIES

[0167] Approximate ambient visual solubility was assessed for Form A, Form B, and Form LC-B crystal forms. Solubilities are calculated based on the total solvent used to give a solution; actual solubilities may be greater because of the volume of the solvent portions utilized or a slow rate of dissolution. Approximate pH of the solutions was measured after solubility determination using indicator stips. The observations are summarized below.

CRYSTALLIZATION EXPERIMENTS AND DATA (A) Summary of Methods of Synthesizing Compound (I) Solid

Forms

[0168] The table below summarizes various conditions employed to prepare the solid forms described herein. "Procedure A" refers to the following procedure: dissolving

Compound (II) in isopropanol, adding acid in EtOH, adding tert-butyl methyl ether until precipitate persists, then slurry the mixture at approximately 42 - 45 °C for the indicated time. SC refers to slow cooling, where a sample is heated to a particular temperature on a hot plate and subsequently cooled by turning off the hot plate.

Seeded Slurry

FORM Approx. Scale Conditions

With Duration

400 mg

VLC - 2 days Procedure A

VLC 400 mg - 6 days Procedure A

400 mg Procedure A, but higher EtOH

LC-B - 1 day

concentration

Procedure A, but higher EtOH

LC-B 400 mg - 4 days

concentration

LC-B 400 mg - slow cool from EtOAc/ EtOH slow cool from IP A/ EtOH,

LC-B 300 mg - - evaporate solvent, slurry in EtOAc

3 days

slow cool from EtOH, evaporate

LC-B 300 mg - - solvent, slurry in ACN 3 days slow cool from EtOAc/ EtOH,

LC-B 300 mg - - slurry 3 days slurry in anhydrous EtOAc, 1 day,

VLC 300 mg - - SC slurry in anhydrous EtOAc, 1 day,

VLC 300 mg A - SC

Procedure A with anhydrous IPA

200 mg

VLC - 1 day (acid also in anhydrous IPA), sub- sample of 3709-67-02

X-ray Procedure A with anhydrous IPA

200 mg - 6 days

amorphous (acid also in anhydrous IPA)

200 mg Procedure A with anhydrous IPA

LC-B A 1 day

(acid also in anhydrous IPA),

Procedure A with anhydrous IPA

LC-B 200 mg A 6 days

(acid also in anhydrous IPA) Seeded Slurry

FORM Approx. Scale Conditions

With Duration

anhydrous IPA/MTBE, heat up to

X-ray

50 mg A - -44 °C, MTBE to cloud point, SC amorphous

with seeding, freezer 1 day

(B) Stability Stressing

[0169] Samples of Form- A, Form-B, Form LC-B, and the amorphous form of Compound (I) were subjected to different levels of relative humidity (R.H.) and temperature, and observations were taken, which are summarized below.

Con¬

Form Time Observations Technique Analysis/ Result ditions

7 dry off-white

- - days solid

dry off-white

solid;

aggregates and

14 crystalline, pattern A, specks, partially XRPD

days minor peak shifts birefringent

(around edges) dry off-white

1 day - - solid

2 dry off-white

- - days solid

dry off-white

7

60% solid - - days

RH/

25 °C dry off-white

solid;

14 aggregates and

- - days specks, partially

birefringent

(around edges)

dry off-white

1 day - - solid

60%

2 dry off-white

B RH/ - - days solid

25 °C

7 dry off-white

- - days solid Con¬

Form Time Observations Technique Analysis/ Result ditions

dry off-white

14 solid;

- - days aggregates, not

birefringent

dry off-white

1 day - - solid

2 dry off-white

- - days solid

75% dry off-white

7

RH/ solid - - days

40 °C

dry off-white

14 solid;

XRPD crystalline pattern B days aggregates, not

birefringent

14 dry yellow low crystalline pattern

60% XRPD

days solids B

RH/

28

25 °C

days

LC-B

14 dry yellow low crystalline pattern

75% XRPD

days solids B

RH/

28

40 °C

days

dry off-white

7 solids, mostly

- - days adhered to

75%

bottom of vial

Amorphous RH/

14

40 °C

days

28

days (C) Drying Experiments

[0170] Drying was performed on Form A, Form B, and Form LC-B. The observations are summarized below.

Form Conditions Observations Technique Analysis/ Result dry,

3.21% weight loss from 25 - -77 - 78 TGA

170 °C

°C, 5 days

heat^a in

capillary to no change in

- -

-88 °C color/ habit continue no change in

heating to color/ habit,

-150 °C, liquid on inside

LC-B hold for of capillary after

~15 min. heating

vacuum

dry, RT,

1 day dry, off-white XRPD

low crystalline pattern B solid

vacuum

dry,

dry dark yellow low crystalline -77 - 82 XRPD

solid pattern B

LC-B °C,

3 days

minor EtOH, EtOAc, and

1H NMR

water present minor MTBE, large amount vacuum

water present, possibly dry, dry, off-white

A 1H NMR broken salt

-79 °C, 1 solid

day Form Conditions Observations Technique Analysis/ Result vacuum

dry, dry off-white X-ray amorphous,

XRPD

-77 - 82 solid peak at -32 °2Θ °C, 3 days

1.06% weight loss from 25 - 170 °C

TG-IR

background water and carbon dioxide greater than volatiles from sample, no HC1 lost

(D) Milling Experiments

[0171] Milling experiments were performed on Form A and LC-B. Observations are summarized in the table below.

Form Solvent Conditions Observations XRPD Result

off-white,

aggregates and

20 min., 30

toluene tiny particles, low crystalline, pattern B

Hz

not

birefringent

(viii) PEAK PICKED XRPD

[0172] The peaks for Form A and Form B XRPD patterns were identified. XRPD patterns with identified peaks are provided in FIG. 15 (Form A) and FIG. 16 (Form B). Peak values and intensity values are provided in the table below. For samples with only one XRPD pattern and no other means to evaluate whether the sample provides a good approximation of the powder average, peak tables contain data identified only as "Prominent Peaks". These peaks are a subset of the entire observed peak list. Prominent peaks are selected from observed peaks by identifying preferably non-overlapping, low-angle peaks, with strong intensity. Although peaks are labeled on diffraction patterns and listed in tables, for technical reasons, different rounding algorithms were used to round each peak to the nearest 0.1° or 0.01° 2Θ, depending upon the instrument used to collect the data and/or the inherent peak resolution. The location of the peaks along the x-axis (° 2Θ) in both the figures and the tables were automatically determined using proprietary software PatternMatch™ 3.0.1 and rounded to one or two significant figures after the decimal point based upon the above criteria. Peak position variabilities are given to within ±0.1° 2Θ based upon recommendations outlined in the USP discussion of variability in x-ray powder diffraction.

FORM A

°2θ d space (A) Intensity (%)

1.54 ± 0.10 57.367 ± 3.984 17

3.25 ± 0.10 27.186 ± 0.863 86

5.74 ± 0.10 15.397 ± 0.273 55

6.61 ± 0.10 13.372 ± 0.205 16

8.80 ± 0.10 10.049 ± 0.115 9

10.33 ± 0.10 8.564 ± 0.083 100

11.44 ± 0.10 7.735 ± 0.068 20

11.86 ± 0.10 7.462 ± 0.063 25 13.30 ± 0.10 6.657 ±0.050 14

14.50 ±0.10 6.109 ±0.042 22

15.13 ± 0.10 5.856 ±0.039 51

15.49 ±0.10 5.721 ±0.037 40

16.51 ±0.10 5.369 ±0.032 66

17.47 ±0.10 5.076 ±0.029 38

18.13 ± 0.10 4.893 ± 0.027 19

19.42 ±0.10 4.571 ±0.023 5

20.20 ±0.10 4.396 ±0.022 36

21.49±0.10 4.135 ±0.019 32

23.05 ±0.10 3.859 ±0.017 40

24.07 ±0.10 3.697 ±0.015 30

24.97 ±0.10 3.566 ±0.014 49

25.87 ±0.10 3.444 ±0.013 15

27.19±0.10 3.280 ±0.012 8

28.42 ±0.10 3.141 ±0.011 8

FORM A Prominent Peaks

°2Θ d space (A) Intensity (%)

3.25 ±0.10 27.186 ±0.863 86

5.74 ±0.10 15.397 ±0.273 55

10.33 ±0.10 8.564 ±0.083 100

16.51 ±0.10 5.369 ±0.032 66

FORM B

°2Θ d space (A) Intensity (%)

I.54 ±0.10 57.367 ±3.984 22

3.25 ±0.10 27.186 ±0.863 10

7.27 ±0.10 12.160 ±0.169 41

9.67 ±0.10 9.147 ±0.095 97

II.26 ±0.10 7.858 ±0.070 80

13.09 ±0.10 6.764 ±0.052 33 14.53 ±0.10 6.096 ± 0.042 58

16.48 ±0. 10 5.379 ± 0.033 9

18.13 ± 0. 10 4.893 ± 0.027 10

18.73 ±0. 10 4.738 ± 0.025 29

20.86 ±0. 10 4.259 ± 0.020 100

22.81 ±0. 10 3.899 ± 0.017 44

24.01 ±0. 10 3.706 ± 0.015 22

26.47 ±0. 10 3.367 ± 0.013 14

28.24 ±0. 10 3.160 ± 0.011 11

FORM B Prominent Peaks

°2Θ d space (A) Intensity (%)

7.27 ±0.10 12.160 ±0.169 41

9.67 ±0.10 9.147 ±0.095 96

11.26 ±0.10 7.858 ±0.070 79

14.53 ±0.10 6.096 ±0.042 57

Pharmacokinetic Data

[0173] Appendix A details experiments detailing pharmacokinetic characteristics of Compound I.

APPENDIX A

GLOSSARY

Tmax Time to reach maximum plasma concentration MATERIALS AND METHODS

IN-LIFE PHASE Test article Compound I Form B ( MW: 809.78 ) and its free base, Compound II (

MW: 736.86 )

Animal SD rats (220-250 g), 6-7 weeks, male, purchased from SLAC

Laboratory Animal Co. LTD.

Qualification No.: SCXK (SH) 2007-0005, 15123

6 rats for oral administration of Compound II and Compound I Form B

Dose Compound I Form B: 19.5 mg/kg equivalent to free base (Compound

II) (10 mL/kg) via oral gavage

Compound II: 18.5 mg/kg (10 mL/kg) via oral gavage

Formulation preparation 0.5% MC (Methylcellulose) in saline

The detailed procedures of formulation preparation were described as follows:

For Compound I Form B:

1 ) Weighed 19.76 mg of Compound I Form B (equivalent to 17.98 mg of free base) into a new tube.

2) Added 1 1.987 mL of 0.5% MC in saline to the tube containing compound.

3) Vortexed the tube for 2-3 min, and sonicated for 15-20 min to obtain a white and homogenous suspension.

For Compound II:

1 ) Weighed 15.16 mg of Compound II into a new tube.

Added 10.107 mL of 0.5% MC in saline to the tube containing compound.

2) Vortexed for 1-2 min, homogenized the mixture by a IKA

homogenizer at 5000 rpm for about 10 min.

3) Vortexed and sonicated the mixture until a homogenous

suspension was obtained.

Administration/Sampling 6 rats for PO administration of Compound I Form B and point Compound II

Pre-dose, 0.083, 0.25, 0.5, 1 , 2, 4, 8, 24 hr, Plasma only,

Fasted over night, free access to water

Fed 4 hr after administration

Blood collection The rats were restrained manually. Approx. 150-200 [it blood/time point was collected into K₂EDTA tubes via tail vein or retro-orbital puncture (under anesthesia with Isoflurane). Blood samples were put on ice and centrifuged (2000g for 5 minutes in 4^°C) to obtain plasma within 30 minutes after collection.

Sample storage and Plasma samples were stored at approximately -70°C until analysis, disposition

QUANTITATION BY LC-MS/MS

LCMSMS-008 (Agilent 6410, triple quadruple)

SD rat plasma

Salt, Compound I Form B, and its free base, Compound II, collectively, "Compound l-N"

Compound Y

Positive ion, ESI

MRM detection

Compound l-ll: [M+2H]²⁺ m/z 369.3→ 70.1

Compound Y: [M+H]⁺ m/z 763.4→ 588.9

Mobile phase A: H₂0- 0.1 %FA-5 mM NH₄OAc

Mobile phase B: MeOH- 0.1 %FA-5 mM NH₄OAc Time

(min) Pump B (%)

0.30 10

0.50 90

2.00 90

2.01 10

3.50 Stop

Column: Waters XBridge Ci₈ (2.1 *50 mm, 3.5 μιη)

Flow rate: 0.40 mL/min

Retention time:

Compound l-ll: 2.31 min;

Compound Y: 2.30 min

For plasma samples:

An aliquot of 30 μΙ_ of plasma sample was added with 30 μΙ_ of the IS (Compound Y, 200 ng/mL) first and partitioned by LLE using 1 mL of Sample preparation MTBE. The mixture was vortexed for 5 min and centrifuged at 10000 rpm for 5 min. An aliquot of 850 μΙ_ of supernatant was dried under N₂ stream. The residue was reconstituted with 150 μΙ_ of 50% MeOH, vortexed for 2 minutes and an aliquot of 5 μΙ_ supernatant was injected onto LC-MS/MS.

For diluted samples:

An aliquot of 10 [it plasma sample was added with 40 [it blank plasma to obtain the diluted plasma samples, and the sample dilution factor is 5. The exaction procedure for diluted samples was the same with non-diluted samples.

Calibration curve 1.00-3000 ng/mL for the Compound l-ll in SD rat plasma

Pharmacokinetic analysis

The PK parameters were determined by using non-compartmental module of WinNonlin®

Professional 5.3. The time points to determine the terminal t_1/2 were selected by the best fit model of WinNonlin. Manual selection of time points would be an alternative method if the best fit model can not determine the terminal phase well. Any cone, data under LLOQ (LLOQ = 1 ng/mL for plasma) were replaced with "BQL".

RESULTS

The concentrations of the Compound l-ll in rat plasma after oral administration of Compound I Form B and Compound II are listed in Tables l-ll, and illustrated in Figures l-lll. Dosing solution analysis is listed in Table III. The PK parameters are listed in Tables IV-VI.

Table I. Individual and mean plasma concentration-time data of Compound l-ll after oral administration of 19.5 mg/kg (equivalent to free base) in Compound I Form B to SD rats

Plasma concentration (ng/mL)

Time(hr) Mean SD CV (%)

Rat#1 Rat#2 Rat#3

0 BQL BQL BQL BQL NA NA

0.083 2233 1733 1583 1850 340 18.4

0.25 9659 5673 7979 7770 2001 25.8

0.5 14413 14522 13649 14195 476 3.35

1 1 1326 15901 13752 13660 2289 16.8

2 13899 17353 8227 13160 4608 35.0

4 9961 13969 12488 12139 2027 16.7

8 4765 9469 6944 7059 2354 33.3

24 35.6 80.4 40.3 52.1 24.6 47.2

Table II. Individual and mean plasma concentration-time data of Compound l-ll after oral administration of 18.5 mg/kg in Compound II to SD rats

Time(hr) Plasma concentration (ng/mL) Mean SD CV (%) Table II. Individual and mean plasma concentration-time data of Compound l-ll after oral administration of 18.5 mg/kg in Compound II to SD rats

Rat#4 Rat#5 Rat#6

0 BQL BQL BQL BQL NA NA

0.083 1882 1610 2276 1923 335 17.4

0.25 6367 6076 6514 6319 223 3.53

0.5 14779 12546 1 1836 13054 1536 1 1.8

1 10975 14487 12885 12782 1758 13.8

2 7131 1 131 1 9713 9385 2109 22.5

4 6650 5831 9625 7369 1996 27.1

8 2730 7563 5750 5348 2441 45.7

24 13.5 36.1 51 .4 33.7 19.0 56.5

Table III. Dosing solution analysis

Measured

Expected Cone. Mean

Sample Name Cone. Accuracy (%)

( g mL) (Mg/mL)

( g mL)

Compound I Form

150 195

B-01

Compound I Form

150 193 195 130

B-02

Compound I Form

150 196

B-03

Compound 11-01 150 188

Compound II-02 150 181 185 124

Compound II-03 150 187

The original dosing solution (1.5 mg/mL) was diluted to be 150 ug/mL (expected Cone.) before dosing solution analysis.

The actual Compound l-ll dosage of 19.5 mg/kg in Compound I Form B and 18.5 mg/kg in Compound II form was used for PK data analysis since the measured dosing concentration was not without 80-120% of the expected concentration.

Figure I. Individual plasma concentration-time profiles of Compound l-ll after oral administration of 19.5 mg/kg (equivalent to free base) in Compound I Form B to SD rats

Figure II. Individual plasma concentration-time profiles of Compound l-ll after oral administration of 18.5 mg/kg in Compound II form to SD rats

8 12 16 20 24

Time (hr) Figure III. Mean plasma concentration-time profiles of Compound l-ll after oral administration of 19.5 mg/kg (equivalent to free base) in Compound I Form B form ("HCI salt") and 18.5 mg/kg in Compound II form ("free base") to SD rats

12 16 20 24

Time (hr)

Table IV. Pharmacokinetic parameters of Compound l-ll in plasma after oral administration of 19.5 mg/kg (equivalent to free base) in Compound I Form B to SD rats

PK parameters Unit Rat#1 Rat#2 Rat#3 Mean SD CV (%)

Tmax hr 0.500 2.00 1.00 1.17 0.764 65.5

Cmax ng/mL 14400 17400 13800 15200 1929 12.7 t-l/2 hr 2.40 2.57 2.34 2.44 0.119 4.90

AUCiast hr^*ng/ml_ 115000 182000 137000 144667 34152 23.6

AUC_INF hr^*ng/ml_ 115000 182000 137000 144667 34152 23.6

Table V. Pharmacokinetic parameters of Compound l-ll in plasma after oral administration of 18.5 mg/kg in Compound II form to SD rats

PK parameters Unit Rat#4 Rat#5 Rat#6 Mean SD CV (%)

Tmax hr 0.500 1.00 1.00 0.833 0.289 34.6

Cmax ng/mL 14800 14500 12900 14067 1021 7.26 t-l/2 hr 2.19 2.68 2.56 2.48 0.255 10.3

AUC_LAST hr^*ng/ml_ 73400 127000 117000 105800 28501 26.9

AUCNF hr^*ng/ml_ 73400 128000 117000 106133 28877 27.2 Table VI. Pharmacokinetic parameters comparison of Compound l-ll in plasma after oral administration in Compound I Form B form and Compound II form to SD rats

Compound I Form B Compound II

PK parameters Unit 19.5 mg/kg equivalent to free base 18.5 mg/kg

Mean SD CV (%) Mean SD CV (%)

Tmax hr 1 .17 0.764 65.5 0.833 0.289 34.6

Cmax ng/mL 15200 1929 12.7 14067 1021 7.26 t-l/2 hr 2.44 0.1 19 4.90 2.48 0.255 10.3

AUCiast hr^*ng/ml_ 144667 34152 23.6 105800 28501 26.9

AUC_INF hr^*ng/ml_ 144667 34152 23.6 106133 28877 27.2

PK profile in SD rat

After an actual oral dose of 19.5 mg/kg (equivalent to free base) in HCI salt form, the C_max value of Compound l-ll in rat plasma was 15200 + 1929 ng/mL, and corresponding mean T_max value was 1.17 + 0.764 hours. The area under curve from time 0 to last time point AUC|_ast and from time 0 to infinity AUCi_NF was 144667 + 34152 and 144667 + 34152 hr^*ng/mL, respectively.

After an actual oral dose of 18.5 mg/kg in free base form, the C_max value of Compound l-ll in rat plasma was 14067 + 1021 ng/mL, and corresponding mean T_max value was 0.833 + 0.289 hours. The area under curve from time 0 to last time point AUCi_ast and from time 0 to infinity AUC_INF was 105800+ 28501 and 106133 + 28877 hr^*ng/mL, respectively.

Standard Curve Summary of Compound l-ll in Rat Plasma

Std 01 Std08

Sample Name Std 02 Std 03 Std 04 Std 05 Std 06 Std 07

(LLOQ) (ULOQ)

Theoretical

1.00 3.00 10.0 30.0 100 300 1000 3000 Cone. ( ng/mL)

Measured ^*0.682 2.69 9.52 32.7 87.2 256 1012 3193

Cone. ( ng/mL)

1.03 ^*3.70 10.2 32.6 101 279 1057 3373

Mean Cone.

1.03 2.69 9.88 32.6 94.0 267 1034 3283 (ng/mL)

Accuracy

103 89.8 98.8 109 94.0 89.1 103 109 (%)

*: The calculated value that was not within 85% to 1 15% (80% to 120% for LLOQ) of the theoretical value was excluded from calibration curve Dilution QC summary of Compound l-ll in rat plasma

Nominal concentration Measured concentration

Sample Name Accuracy (%)

DQC-02 480 523 109

DQC-03 480 527 1 10

Claims

WHAT IS CLAIMED:

1. A solid form comprising the compound of formula (I):

2. The solid form of claim 1, wherein the solid form is the Form B crystal form of the compound of formula (I).

3. The solid form of claim 1, wherein the solid form is the Form LC-B crystal form of the compound of formula (I).

4. The solid form of claim 1 having an X-ray powder diffraction pattern comprising peaks at approximately 9.67, 11.26, 18.73, and 20.86 degrees 2Θ.

5. The solid form of claim 4 having an X-ray powder diffraction pattern further comprising peaks at approximately 7.27, 14.53 and 24.01 degrees 2Θ.

6. The solid form of claim 1 having an X-ray powder diffraction pattern which matches the XRPD pattern presented in FIG 1 or FIG 2..

7. The solid form of claim 1 having a differential scanning calorimetry thermogram comprising an endothermic event with a peak maximum of approximately 82 °C.

8. The solid form of claim 1 having a differential scanning calorimetry thermogram which matches the differential scanning calorimetry thermogram presented in FIG 3.

9. The solid form of claim 1 having a thermal gravimetric analysis curve that exhibits a weight loss of about 4% of the total sample weight between about 25 - 170 °C.

10. The solid form of claim 1 having a thermal gravimetric analysis curve which matches the thermal gravimetric analysis curve presented in FIG 4.

11. The solid form of claim 1 , wherein the solid form is the Form A crystal form of the compound of formula (I).

12. The solid form of claim 1 having an X-ray powder diffraction pattern comprising peaks at approximately 3.25, 5.74, 10.33, 16.51 degrees 2Θ.

13. The solid form of claim 17 having an X-ray powder diffraction pattern further comprising peaks at approximately 15.13, 17.47, 23.05, and 24.97 degrees 2Θ.

14. The solid form of claim 1 having an X-ray powder diffraction pattern which matches the XRPD pattern presented in FIG 5.

15. The solid form of claim 1 having a differential scanning calorimetry thermogram comprising an endothermic event with a peak maximum of approximately 82 °C and approximately 216 °C.

16. The solid form of claim 1 having a differential scanning calorimetry thermogram which matches the differential scanning calorimetry thermogram presented in FIG 6.

17. The solid form of claim 1 having a thermal gravimetric analysis curve which matches the thermal gravimetric analysis curve presented in FIG 7.

18. A pharmaceutical composition comprising a solid form of any of claims 1-17.

19. The pharmaceutical composition of claim 18 further comprising one or two additional compounds having anti-HCV activity.

20. A method of treating HCV infection in a mammal comprising administering to the mammal a therapeutically effective amount of the solid form of any of claims 1-17 or the pharmaceutical composition of claim 18 or 19.

21. A method for producing a crystal form of the compound of formula (I):

(I) comprising the steps of

(a) contacting the compound of formula (II):

(Π)

with about 2 molar equivalents of HC1 in the presence of ethanol and contacting resulting reaction mixture of step (a) with tert-butyl methyl ether to form a slurry; and

heating the slurry of step (b) to about 42 °C.

A method for producing a crystal form of the compound of formula (I)

(I) comprising the steps of: contacting the compound of formula (II)

(Π)

with HCl in the presence of ethanol at 40 - 60 °C to form a mixture;

(b) heating the mixture for 1 - 6 hours at 40 - 60 °C;

(c) concentrating the mixture to dryness to obtain a solid;

(d) heating the solid of step (c) in refluxing methanol for 3 - 8 hours in the presence of activated carbon;

(f) heating the concentrated filtrate of step (e) in the presence of acetone at 40

- 60 °C for 12 - 36 hours and subsequently cooling to room temperature to obtain a suspension;

(g) stirring the suspension of step (f) for 12 - 36 hours; and

(h) filtering the suspension.

23. A method for producing a compound of formula (II):

(Π) comprising contacting the compound of formula A-l 1 :

A-l l

or a salt thereof, with (5)-N-methoxycarbonyl-valine in the presence of peptide coupling reagent or reagents.

24. The method of claim 33, wherein the peptide couple reagent or reagents is selected from the group consisting of:

(a) 2-(7~Aza~ 1 H-benzotriazole- 1 -yl)- 1 ,1 ,3 ,3~l^'etrarnethyluroxiium

liexafluorophosphate;

(b) dicyclohexylcarbodiimide;

(d) benzotriazol- 1 -yl-oxytripyrrolidinophosphonium hexafluorophosphate;

(e) 4-(4,6-dimethoxy-l,3,5-triazin-2-yl)-4-methylmorpholinium chloride;

(f) 3 -(3 -dimethylaminopropyl)carbodiimide;

(g) hydroxybenzotriazole; and

(h) l-ethyl-3-(3-dimethylaminopropyl) carbodiimide and hydroxybenzotriazole.

25. A method for producing a compound of formula A-l l :

A-l l

A-10

wherein PG is a protecting group. A method for producing the compound of formula A- 10

A-10 comprising contacting the compound of formula A-9

A-9

with the compound of formula A-

27. A method for producing the compound of formula A-10:

A-10

comprising contacting the compound of formula A-8:

A-8 with the compound of formula A-5

A-5

28. A method for producing the compound of formula A-9:

A-9

comprising

(a) contacting the compound of formula A-8:

A-8

with ethynyltrimethylsilane in the presence of piperidine, Pt-Bu₃, Cul, and

Pd(PPh₃)₂Cl₂ to form an intermediate ; and

contacting the intermediate of step (a) with K₂C0₃ in the presence of methanol.

A method for producing the compound of formula B-8

B-8 comprising contacting the compound of formula B-7:

B-7

with a protecting group cleaving agent, wherein PG is a protecting group.

A method for producing the compound of formula B-7

B-7

comprising contacting the compound of formula A-8:

with the compound of formula B-2:

B-2 in the presence of a coupling reagent or reagents, wherein X is Br, I, or OH and PG is a protecting group.

31. A method for producing the compound of formula B-6:

B-6

comprising contacting the compound of formula B-5 :

B-5

a protecting group cleaving agent, wherein PG is a protecting group.

32. A method for producing the compound of formula B-5 :

B-5 comprising contacting the compound of formula A-2:

A-2

with the compound of formula B-4:

33. A method for preparing the compound of formula C-2:

comprising compound the compound of formula A-9:

with the compound of formula C-l :

C-l

34. A method for producing the compound of formula C-3 :

C-3

comprising contacting the compound of formula B-4:

B-4

with the compound of formula C-l :

C-l

in the presence of a coupling reagent or reagents, wherein PG is a protecting group and X is

Br, I, or OH.

A method for producing the compound of formula D-2

D-2

comprising contacting the compound of formula A-5 :

A-5 with the compound of formula D-l :

D-l

Br, I, or OH.

A method for producing the compound of formula D-3

D-3 comprising contacting the compound of formula B-2

B-2

with the compound of formula D-l :

D-l

Br, I, or OH.