TW202214572A - Synthesis of substituted arylmethylureas, analogues, and crystalline forms thereof and methods of using same - Google Patents

Abstract

The present disclosure includes synthetic methods for preparing certain substituted (hetero)arylmethyl urea compounds, compositions comprising the same, and crystalline forms thereof, which can be used to treat, ameliorate, and/or prevent hepatitis B virus (HBV) infections in a patient.

Description

Synthesis of Substituted Aryl Methyl Ureas, Analogs and Crystal Forms and Methods of Using the Same

This case claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 63/121,021, filed December 3, 2020, and U.S. Provisional Patent Application No. 63/040,211, filed June 17, 2020, all of which It is incorporated herein by reference in its entirety.

This case relates to the synthesis of substituted aryl methyl ureas, their analogs and crystalline forms, and methods of using them.

Hepatitis B is one of the most prevalent diseases in the world and is listed as a high priority area of concern by the National Institute of Allergy and Infectious Diseases (NIAID). While most people clear up the infection after acute symptoms, about 30 percent of cases become chronic. An estimated 350-400 million people worldwide suffer from chronic hepatitis B, causing 50-1 million deaths each year, mainly due to the development of hepatocellular carcinoma, cirrhosis and/or other complications.

A limited number of drugs are currently approved for the treatment of chronic hepatitis B, including two alpha-interferon preparations (standard and pegylated) that inhibit HBV DNA polymerase and five nucleoside/nucleotide analogs ( lamivudine, adefovir, entecavir, telbivudine and tenofovir). Currently, the first-line treatment options are entecavir, tenofovir, or peginterferon alfa-2a. However, peginterferon alfa-2a achieves the desired serological milestone in only one third of treated patients, often with severe side effects. Entecavir and tenofovir require long-term or possibly lifelong dosing for sustained suppression of HBV replication and may eventually fail due to the emergence of resistant viruses. Therefore, there is an urgent need to introduce novel, safe, and effective treatments for chronic hepatitis B.

Hepatitis B is caused by the Hepatitis B virus (HBV), a non-cytopathic hepadnavirus that belongs to the family Hepadnaviridae . Pregenomic (pg) RNA is the template for reverse transcription replication of HBV DNA. Encapsidation of pg RNA into the nucleocapsid together with viral DNA polymerase is critical for subsequent viral DNA synthesis. Inhibition of pg RNA encapsidation blocks HBV replication and provides a new therapeutic avenue for HBV treatment. Capsid inhibitors work by directly or indirectly inhibiting the expression and/or function of capsid proteins: for example, it can inhibit capsid assembly, induce the formation of non-capsid polymers, promote excessive capsid assembly or misdirected capsid Shell assembly, affect capsid stability, and/or inhibit RNA encapsidation. Capsid inhibitors can also act by inhibiting capsid function during one or more downstream events during replication, such as, but not limited to, viral DNA synthesis, transport of relaxed circular DNA (rcDNA) to the nucleus, co-coding Valence closed circular DNA (cccDNA) formation, virus maturation, budding and/or release.

Hepatitis D virus (HDV) is a small circular enveloped RNA virus that reproduces only in the presence of HBV. Specifically, HDV requires the HBV surface antigen protein to reproduce itself. Infection with HBV and HDV leads to more serious complications than infection with HBV alone. These complications include a greater likelihood of experiencing liver failure with acute infection and rapid progression to cirrhosis, and an increased chance of developing liver cancer with chronic infection. When combined with hepatitis B, hepatitis D has the highest mortality rate of all hepatitis infections. The transmission route of HDV is similar to that of HBV. Infection is largely restricted to groups at high risk of HBV infection, particularly injecting drug users and those receiving clotting factor concentrates.

In the clinic, inhibition of pg RNA encapsidation or, more generally, nucleocapsid assembly may offer certain therapeutic advantages for the treatment of hepatitis B and/or D. In one aspect, inhibition of pgRNA encapsidation can complement current drugs by providing options to patient subpopulations who cannot tolerate or benefit from current drugs. On the other hand, based on its unique antiviral mechanism, inhibition of pg RNA encapsidation is effective against HBV and/or HDV variants that are resistant to currently available DNA polymerase inhibitors. On the other hand, the combination therapy of pg RNA encapsidation inhibitor and DNA polymerase inhibitor can synergistically inhibit HBV and/or HDV replication and prevent the emergence of drug resistance, thus chronic hepatitis B and/or hepatitis D infection Provides more effective treatment.

Currently, there is no effective antiviral therapy available for the treatment of acute or chronic hepatitis D. Interferon-alpha given weekly for 12 to 18 months is the only licensed therapy for hepatitis D. Responses to this therapy have been limited, as only about a quarter of patients have undetectable serum HDV RNA 6 months after treatment.

Accordingly, there is a need in the art to identify novel compounds that can be used to treat and/or prevent HBV and/or HDV infection in a subject. In certain embodiments, the novel compounds inhibit HBV and/or HDV nucleocapsid assembly. In other embodiments, the novel compounds are useful in patients infected with HBV and/or HBV-HDV, patients at risk for infection with HBV and/or HBV-HDV, and/or patients infected with drug-resistant HBV and/or HDV . The present invention addresses this need.

(X) In one aspect, the present disclosure includes the preparation of ( R )-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxo-1,2- Dihydroisoquinolin-4-yl)ethyl)-1-methylurea (also known as Compound (X)), or its salts, solvates, prodrugs, isotopically labeled derivatives, stereoisomers ( such as, in a non-limiting example, an enantiomer or any mixture thereof, such as, in a non-limiting example, a mixture of its enantiomers in any ratio), and/or a tautomer, and Method for any mixture:

(X)

In another aspect, the present disclosure provides developable forms of (X) or a solvate thereof that are useful for treating, ameliorating and/or preventing HBV (and/or HBV-HDV) infection and related conditions in a subject. In other embodiments, the present disclosure provides polymorphs of (X) , which may include any solvates thereof.

In certain aspects, the present disclosure relates to the discovery of scalable synthetic routes that allow for the reproducible multigram synthesis of certain substituted urea-containing compounds. In other aspects, the present disclosure relates to developable forms of certain substituted urea compounds useful in the treatment, amelioration and/or prevention of HBV (and/or HBV-HDV) infection and related symptoms in a subject.

(X) In one aspect, the present disclosure includes the preparation of ( R )-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxo-1,2- Dihydroisoquinolin-4-yl)ethyl)-1-methylurea (also known as Compound (X)), or its salts, solvates, prodrugs, isotopically labeled derivatives, stereoisomers ( such as, in a non-limiting example, an enantiomer or any mixture, such as, in a non-limiting example, a mixture of its enantiomers in any ratio), and/or tautomers, and their Method for any mixture:

(X)

In another aspect, the present disclosure provides developable forms of (X) or a solvate thereof. In other embodiments, the present disclosure provides polymorphs of (X) (which may include any solvates thereof).

Certain crystalline polymorphic forms of (X) are described herein. During the crystallization screening, from materials such as acetone, tetrahydrofuran, ethanol, dichloromethane, dimethylformamide/water, DMF/tert-butyl methyl ether, water, tetrahydrofuran, acetone/water, 2-methyltetrahydrofuran, 2- Solvents such as propanol, n-heptane, methanol, toluene, dimethylsulfoxide/methyl ethyl ketone/toluene, ethyl acetate, and tetrahydrofuran/water identify certain polymorphic forms. Solvates including ethanol, ethyl acetate, methanol, possibly mixed solvates/hydrates or acetone solvates, and possibly non-stoichiometric hydrates (forms 4, 5, 6, 7 and 9) and unsolvated forms (forms 3 and 8). The dichloromethane solvate (Form 1) and the stable unsolvated form (Form 2) were identified prior to this screening.

PCT International Application PCT/US2019/065756 filed on December 11, 2019 (published as WO 2020/123674 on June 18, 2020), US Provisional Application 62/896,237 filed on September 5, 2019, and December 2018 The disclosure of US Provisional Application 62/778,471, filed 12, is incorporated herein by reference in its entirety.

definition

Throughout this specification, when a composition is described as having, comprising or comprising particular components, or when a process is described as having, comprising or comprising particular process steps, it is contemplated that the composition of the present teaching also consists essentially of the recited The components consist of or consist of, and the processes of the present teachings also consist essentially of or consist of the enumerated processing steps.

In this application, when an element or component is referred to as being included in and/or selected from a list of recited elements or components, it will be understood that the element or component may be among the recited elements or components and may be selected from two or more of the listed elements or components.

As used herein, unless otherwise defined, all technical and scientific terms generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature and laboratory procedures in organic chemistry, virology, biochemistry and pharmaceutical sciences used herein are those well known and commonly used in the art. Also, it is to be understood that the phraseology or terminology employed herein and not otherwise defined is for the purpose of description and not of limitation. The use of any section headings is intended to aid reading of the document and should not be construed as limiting; information related to section headings may appear within or outside of that particular section. All publications, patents, and patent documents mentioned in this document are incorporated by reference in their entirety, as if individually incorporated by reference.

In the methods described herein, actions can be performed in any order unless a time or sequence of operations is explicitly stated. Furthermore, specified actions may be performed concurrently unless explicit declarative language states that they are performed separately. For example, a claimed X action and a claimed Y action may be performed simultaneously in a single operation, and the resulting process would fall within the literal scope of the claimed process.

As used herein, the terms "a," "an," or "the" are used to include one or more unless the context clearly dictates otherwise. Unless stated otherwise, the term "or" is used to mean a non-exclusive "or". The statement "at least one of A and B" or "at least one of A or B" has the same meaning as "A, B or A and B".

As used herein, the term "about" will be understood by one of ordinary skill in the art and will vary to some extent in the context in which it is used. As used herein, "about" when referring to a measurable value such as an amount, a duration of time, etc., is intended to encompass ±20%, ±10%, ±5%, ±1%, or ±0.1% of the specified value. % variation as such variation is suitable for performing the disclosed method.

As used herein, unless otherwise specified, the term "alkenyl", alone or in combination with other terms, refers to a stable monounsaturated or diunsaturated straight or branched chain hydrocarbon group having the specified number of carbon atoms. Examples include vinyl, propenyl (or allyl), crotyl, isopentenyl, butadienyl, 1,3-pentadienyl, 1,4-pentadienyl, and more High homologs and isomers. An example of a functional group representing an alkene is -CH ₂ -CH=CH ₂ .

As used herein, unless otherwise specified, the term "alkoxy," used alone or in combination with other terms, refers to a specified number of carbons, as defined elsewhere herein, attached to the remainder of the molecule via an oxygen atom. Alkyl groups of atoms such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (or isopropoxy) and higher homologues and isomers. Specific examples are (C ₁ -C ₃ )alkoxy groups such as, but not limited to, ethoxy and methoxy.

As used herein, unless otherwise stated, the term "alkyl" by itself or as part of another substituent refers to a group having the specified number of carbon atoms (ie, _C1 - _C10 represents 1 to 10 carbon atoms). straight or branched chain hydrocarbons, and includes straight, branched or cyclic substituents. Examples include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, and cyclopropylmethyl. Specific embodiments are (C ₁ -C ₆ )alkyl groups such as, but not limited to, ethyl, methyl, isopropyl, isobutyl, n-pentyl, n-hexyl, and cyclopropylmethyl.

As used herein, unless otherwise specified, the term "alkynyl," used alone or in combination with other terms, refers to a stable straight or branched chain hydrocarbon group having the specified number of carbon atoms having a carbon-carbon triple bond. Non-limiting examples include ethynyl and propynyl, as well as higher homologs and isomers. The term "propargyl" refers to a group exemplified by _-CH2 -C≡CH. _The term " _{homopropargylic} " refers to a group exemplified by -CH2CH2-C≡CH.

As used herein, the term "aromatic" refers to having one or more polyunsaturated rings and having aromatic character, ie having (4n+2) delocalized π(pi) electrons (where "n" is Integer) of the carbocyclic or heterocyclic ring.

As used herein, unless otherwise specified, the term "aryl" used alone or in combination with other terms refers to a carbocyclic aromatic group containing one or more rings (usually one, two or three rings). systems in which the rings can be linked together in a pendant fashion, such as biphenyl, or can be fused, such as naphthalene. Examples include phenyl, anthracenyl, and naphthyl. Aryl also includes, for example, a benzene or naphthalene ring fused to one or more saturated or partially saturated carbocyclic rings (eg, bicyclo[4.2.0]octa-1,3,5-trienyl or indanyl) , which may be substituted at one or more carbon atoms of an aromatic ring and/or a saturated or partially saturated ring.

As used herein, the term "aryl-( _C1 - _C6 )alkyl" refers to a functional group in which a ₁ to ₆ carbon alkylene chain is attached to an aryl group, eg, -CH2CH2-phenyl or _-CH2 -phenyl (or benzyl). Specific examples are aryl-CH2- and aryl-CH( _{CH3 )} -. The term "substituted aryl-( _C1 - _C6 )alkyl" refers to an aryl-( _C1 - _C6 )alkyl functional group in which the aryl group is substituted. A specific example is substituted aryl ( _CH2 )-. Similarly, the term "heteroaryl-( _C1 - _C6 )alkyl" refers to a functional group in which a ₁ to ₃ carbon alkylene chain is attached to a heteroaryl, eg, -CH2CH2-pyridyl. A specific example is heteroaryl-( _CH2 )-. The term "substituted heteroaryl-( _C1 - _C6 )alkyl" refers to a heteroaryl-( _C1 - _C6 )alkyl functional group in which the heteroaryl group is substituted. A specific example is substituted heteroaryl-( _CH2 )-.

For the purposes of the present invention, the terms "compound," "analog," and "composition of matter" apply equally to the prodrug agents described herein, including enantiomeric forms, diastereomeric forms, salts, and the like , and the terms "compound," "analog," and "composition of matter" are used interchangeably throughout this specification.

As used herein, unless otherwise specified, the term "cycloalkyl" by itself or as part of another substituent refers to a cyclic chain hydrocarbon having the specified number of carbon atoms (ie, C3 _{- C6} refers to a group consisting of cyclic group consisting of 3 to 6 carbon atoms) and includes straight chain, branched chain or cyclic substituents. Examples of (C3 _{- C6} )cycloalkyl are cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Cycloalkyl rings can be optionally substituted. Non-limiting examples of cycloalkyl include: cyclopropyl, 2-methyl-cyclopropyl, cyclopropenyl, cyclobutyl, 2,3-dihydroxycyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, decahydronaphthyl, 2,5-dimethylcyclopentyl, 3,5-dichlorocyclo Hexyl, 4-hydroxycyclohexyl, 3,3,5-trimethylcyclohex-1-yl, octahydropentenyl, octahydro-1 H -indenyl, 3a,4,5,6,7,7a - Hexahydro- 3H -inden-4-yl, decahydroazulene; bicyclo[6.2.0]decyl, decahydronaphthyl and dodecahydro- 1H -fluorenyl. The term "cycloalkyl" also includes bicyclic hydrocarbon rings, non-limiting examples of which include bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.1]heptyl, 1, 3-Dimethyl[2.2.1]heptan-2-yl, bicyclo[2.2.2]octyl and bicyclo[3.3.3]undecanyl.

As used herein, a "disease" is a state of health in a subject in which the subject is unable to maintain homeostasis, and if the disease does not improve, the subject's health continues to deteriorate.

As used herein, a "disorder" in a subject is a state of health in which the subject is able to maintain homeostasis, but in which the state of health of the subject is less favorable than it would be in the absence of the disorder. If left untreated, the condition will not necessarily lead to a further decline in the subject's health status.

As used herein, an "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" of a compound is an amount of the compound sufficient to provide a beneficial effect to the subject to which the compound is administered.

As the term is used herein, "instructional material" includes publications, records, diagrams, or any other presentation medium that can be used to convey the usefulness of the compositions and/or compounds of the invention in a kit. The kit's instructional material, for example, can be affixed to the container containing the compound and/or composition of the present invention or shipped with the container containing the compound and/or composition. Alternatively, the instructional material may be shipped separately from the container for the recipient to use the instructional material with the compound. Delivery of the instructional material may, for example, be physical delivery by means of a publication or other medium of expression that conveys the usefulness of the set, or may alternatively be effected by electronic transmission, for example, by computer means such as by electronic mail , or download from the website.

As used herein, the term "halide" refers to a negatively charged halogen atom. Halide anions are fluoride (F ⁻ ), chloride (Cl ⁻ ), bromide (Br ⁻ ) and iodide (I ⁻ ).

As used herein, unless otherwise indicated, the terms "halo" or "halogen" alone or as part of another substituent refer to a fluorine, chlorine, bromine or iodine atom.

As used herein, unless otherwise specified, the term "heteroalkenyl" by itself or in combination with another term refers to a stable compound consisting of the specified number of carbon atoms and one or two heteroatoms selected from O, N, and S. A linear or branched mono- or di-unsaturated hydrocarbon group in which the nitrogen and sulfur atoms may be optionally oxidized, and the nitrogen heteroatom may be optionally quaternary aminated. Up to two heteroatoms can be placed in succession. Examples include -CH=CH-O- _CH3 , -CH=CH- _CH2 -OH, _-CH2 -CH=N- _OCH3 , -CH=CH-N( _CH3 ) _-CH3 and _-CH2 -CH=CH- _CH2 -SH.

As used herein, unless otherwise specified, the term "heteroalkyl" by itself or in combination with another term refers to a group consisting of the specified number of carbon atoms and one or two heteroatoms selected from O, N, and S. A stable straight or branched chain alkyl group consisting of, and wherein nitrogen and sulfur atoms can be optionally oxidized, and nitrogen heteroatoms can optionally be quaternary ammonium. The heteroatom(s) can be located anywhere in the heteroalkyl group, including between the remainder of the heteroalkyl group and the fragment to which it is attached, and to the most distal carbon atom in the heteroalkyl group. _Examples include _{: -OCH2CH2CH3} , _-CH2CH2CH2OH , _{-CH2CH2NHCH3 , -CH2SCH2CH3} , _{and -CH2CH2S ( = O ) CH3 .} Up to _two heteroatoms may be consecutive, such as, for example, _{-CH2NH - OCH3} or _-CH2CH2SSCH3 .

As used herein, the term "heteroaryl" or "heteroaromatic" refers to a heterocyclic ring having aromatic character. Polycyclic heteroaryl groups may include one or more partially saturated rings. Examples include tetrahydroquinoline and 2,3-dihydrobenzofuranyl.

As used herein, unless otherwise stated, the terms "heterocycle" or "heterocyclyl" or "heterocyclic" by themselves or as part of another substituent are meant to include carbon atoms and are selected from the group consisting of N, O, and An unsubstituted or substituted, stable monocyclic or polycyclic heterocyclic ring system of at least one heteroatom of S, and wherein the nitrogen and sulfur heteroatoms may be optionally oxidized, and the nitrogen atom may be optionally quaternary aminated . Unless otherwise specified, the heterocyclic ring system can be attached to any heteroatom or carbon atom that provides a stable structure. Heterocycles can be aromatic or non-aromatic in nature. In certain embodiments, the heterocycle is a heteroaryl.

Examples of non-aromatic heterocycles include monocyclic groups such as aziridine, ethylene oxide, thiirane, azetidine, oxetane, thietane, pyrrole pyridine, pyrroline, imidazoline, pyrazolidine, dioxolane, cyclobutane, 2,3-dihydrofuran, 2,5-dihydrofuran, tetrahydrofuran, tetrahydrothiophene (thiophane), oxidine, 1 , 2,3,6-tetrahydropyridine, 1,4-dihydropyridine, tetrahydropyridine, morpholine, thiomorpholine, pyran, 2,3-dihydropyran, tetrahydropyran, 1,4 -Dioxane, 1,3-Dioxane, Homopyridine, Homopyridine, 1,3-dioxane, 4,7-dihydro-1,3-dioxepin ) and epoxy hexane (hexamethyleneoxide).

Examples of heteroaryl groups include pyridyl, pyridyl, pyrimidinyl (such as, but not limited to, 2- and 4-pyrimidinyl), pyridyl, thienyl, furyl, pyrrolyl, imidazolyl, thiazolyl, ethylene azolyl, pyrazolyl, isothiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-triazolyl, tetrazolyl, 1,2,3 -thiazolyl, 1,2,3-oxadiazolyl, 1,3,4-thiazolyl and 1,3,4-oxadiazolyl.

Examples of polycyclic heterocycles include indolyl (such as, but not limited to, 2-, 3-, 4-, 5-, 6-, and 7-indolyl), indolinyl, quinolinyl, tetrahydroquinoline olinyl, isoquinolinyl (such as, but not limited to, 1- and 5-isoquinolinyl), 1,2,3,4-tetrahydroisoquinolinyl, cinnolinyl, quinolinyl (eg, but not limited to, 2- and 5-quinoxolinyl), quinazolinyl, phthaloyl, 1,8-naphthyridinyl, 1,4-benzodiethylalkyl, coumarin, di Hydrocoumarin, 1,5-naphthyridinyl, benzofuranyl (eg, but not limited to, 3-, 4-, 5-, 6-, and 7-benzofuranyl), 2,3-dihydrobenzene furanyl, 1,2-benzoxazolyl, benzothienyl (such as, but not limited to, 3-, 4-, 5-, 6-, and 7-benzothienyl), benzoxazolyl, benzothiazolyl (such as, but not limited to, 2-benzothiazolyl and 5-benzothiazolyl), purinyl, benzimidazolyl, benzotriazolyl, thioxanthinyl, oxazole group, quinolinyl, acridinyl, pyrrolizidinyl and quinolizidinyl.

The above list of heterocyclyl and heteroaryl moieties is intended to be representative, not limiting.

As used herein, the term "NMT" means not exceeding.

As used herein, the term "pharmaceutical composition" or "composition" refers to a mixture of at least one compound useful in the present invention and a pharmaceutically acceptable carrier. Pharmaceutical compositions facilitate administration of a compound to a subject.

As used herein, the term "pharmaceutically acceptable" refers to a relatively non-toxic material, such as a carrier or diluent, that does not abrogate the biological activity or properties of the compounds useful in the present invention, ie, can be used without causing adverse biological effects or administer the material to the subject without interacting in a deleterious manner with any component of the composition in which it is contained.

As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable carrier that participates in carrying or transporting a compound useful in the present invention in or to a subject so that it can perform its intended function materials, compositions or carriers such as liquid or solid fillers, stabilizers, dispersants, suspending agents, diluents, excipients, thickeners, solvents or encapsulating materials. Typically, such constructs are carried or transported from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation, including the compounds useful in the present invention, and not injurious to the subject. Some examples of materials that can be used as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as cornstarch and potato starch; cellulose and derivatives thereof, such as sodium carboxymethylcellulose , ethyl cellulose and cellulose acetate; powdered gum tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, Corn oil and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol and polyethylene glycols; esters such as ethyl oleate and ethyl laurate; agar; buffers such as hydrogen Magnesium oxide and aluminum hydroxide; surfactants; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; phosphate buffered solution; and other nontoxic compatible substances used in pharmaceutical preparations.

As used herein, "pharmaceutically acceptable carrier" also includes any and all coatings, antibacterial and antifungal, that are compatible with the activity of the compounds useful in the present invention and are physiologically acceptable to the subject agents, and absorption delaying agents. Supplementary active compounds can also be incorporated into the compositions. A "pharmaceutically acceptable carrier" may further include pharmaceutically acceptable salts of the compounds useful in the present invention. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the present invention are known in the art and are described, for example, in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA). description, which is incorporated herein by reference.

As used herein, the language "pharmaceutically acceptable salts" refers to pharmaceutically acceptable salts formed from inorganic acids, inorganic bases, organic acids, inorganic bases, solvates (including hydrates) and clathrates thereof. Salts of the administered compounds are prepared from accepted non-toxic acids and/or bases.

As used herein, a "pharmaceutically effective amount", "therapeutically effective amount" or "effective amount" of a compound is an amount of the compound sufficient to provide a beneficial effect to the subject to which the compound is administered.

As used herein, the terms "prevent," "prevent," and "avoid" refer to avoiding or delaying symptoms associated with a disease or disorder in subjects who have not developed such symptoms at the time of initiation of administration of an agent or compound outbreak. Disease, disorder and symptom are used interchangeably herein.

As used herein, a "patient" or "subject" can be a human or non-human mammal or bird. Non-human mammals include, for example, livestock and pets such as sheep, cattle, porcine, canine, feline and murine mammals. In certain embodiments, the subject is a human.

The term "specifically binds" or "specifically binds" as used herein means that a first molecule binds preferentially, but not necessarily only, to a second molecule (eg, a particular receptor or enzyme).

As used herein, the terms "subject" and "individual" and "patient" are used interchangeably and can refer to a human or non-human mammal or bird. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline, and murine mammals. In certain embodiments, the subject is a human.

As used herein, the term "substituted" means that an atom or group of atoms has been replaced with hydrogen as a substituent attached to another group.

As used herein, the term "substituted alkyl", "substituted cycloalkyl", "substituted alkenyl" or "substituted alkynyl" refers to an alkyl group as defined elsewhere herein , cycloalkyl, alkenyl or alkynyl independently selected from halogen, -OH, alkoxy, tetrahydro-2-H-pyranyl, _-NH2 , -NH( _{C1 -} C6alkane base), -N(C ₁ -C ₆ alkyl) ₂ , 1-methyl-imidazol-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-4-yl, -C(=O )OH, -C(=O)O(C ₁ -C ₆ ) alkyl, trifluoromethyl, -C≡N, -C(=O)NH ₂ , -C(=O)NH(C ₁ - C ₆ ) alkyl, -C(=O)N((C ₁ -C ₆ ) alkyl) ₂ , -SO ₂ NH ₂ , -SO ₂ NH(C ₁ -C ₆ alkyl), -SO ₂ N (C ₁ -C ₆ alkyl) ₂ , -C(=NH)NH ₂ and -NO ₂ are substituted with one, two or three substituents, in certain embodiments containing, in certain embodiments, independently selected from halogen, -OH , alkoxy, _-NH2 , trifluoromethyl, -N( _CH3 ) ₂ and one or two substituents of -C(=O)OH, in certain embodiments independently selected from halogen, alkane Oxygen and -OH. Examples of substituted alkyl groups include, but are not limited to, 2,2-difluoropropyl, 2-carboxycyclopentyl, and 3-chloropropyl.

For aryl, aryl-( _{C1 -} C3)alkyl and heterocyclyl, the term "substituted" applied to the rings of these groups refers to any level of substitution where such substitution is permissible, That is, single, two, three, four or five substitutions. Substituents are independently selected, and the substitution can be at any chemically accessible position. In certain embodiments, the number of substituents varies between 1 and 4. In other embodiments, the number of substituents varies between 1 and 3. In another embodiment, the number of substituents varies between 1 and 2. In yet other embodiments, the substituents are independently selected from _C1 - _C6 alkyl, -OH, _C1 - _C6 alkoxy, halogen, cyano, amino, acetamido, and nitro. As used herein, when a substituent is an alkyl or alkoxy group, the carbon chain can be branched, straight or cyclic.

Unless otherwise stated, when ^two substituents are taken together to form a ring with the specified number of ring atoms (eg, R and ^R together with the nitrogen to which they are attached form a ring with 3 to 7 ring members), the ring Can have carbon atoms and optionally one or more (eg, 1 to 3) additional heteroatoms independently selected from nitrogen, oxygen, or sulfur. The ring may be saturated or partially saturated, and may be optionally substituted.

Whenever a term or any of its prefix roots appears in the name of a substituent, that name should be construed to include those limitations provided herein. For example, whenever the terms "alkyl" or "aryl" or any of their prefix roots appear in the name of a substituent (eg, arylalkyl, alkylamino), that name should be construed as including herein Those restrictions given elsewhere for "alkyl" and "aryl" respectively.

In certain embodiments, substituents of compounds are disclosed in groups or ranges. It is specifically intended that this description include each and each individual subcombination of the members of these groups and ranges. For example, the term "C _1-6 alkyl" is expressly intended to disclose C ₁ , C ₂ , C ₃ , C ₄ , C ₅ , C ₆ , C ₁ -C ₆ , C ₁ -C ₅ , C ₁ - _C4 , _{C1 -} C3, _{C1 -} C2, C2 _{- C6} , C2 _{- C5} , _{C2 - C4} , C2 _- C3, C3 _{- C6} , C3 _{- C5} , C ₃ -C ₄ , C ₄ -C ₆ , C ₄ -C ₅ and C ₅ -C ₆ alkyl groups.

The terms "treating," "treating," and "treating," as used herein, refer to reducing the frequency or severity of symptoms of a disease or disorder experienced by a subject by administering an agent or compound to the subject.

The term "vol" as used herein to describe the amount of solvent (where no specified volume is provided) refers to 1 L of solvent per 100 g of limiting reagent. For example, in the context of describing "dissolving compound A (100 g) in water (10 vol)", the amount of water used to dissolve compound A is 1 L.

The term "V" as used herein to describe the amount of solvent (where no specified volume is provided) refers to 1 L of solvent per 1 kg of limiting reagent. For example, in the context of the description "Compound A (100 g) is dissolved in water (10 V)", the amount of water used to dissolve Compound A is 100 mL (0.1 L).

Some abbreviations used herein are as follows: cccDNA, covalently closed circular DNA; CPME, cyclopentylmethane; DMF, dimethylformamide; DMSO, dimethylsulfoxide; DNA, deoxyribonucleic acid; DSC, differential Scanning calorimetry; GC, gas chromatography; GPC, gel permeation chromatography; HBsAg, HBV surface antigen; HBV, hepatitis B virus; HDV, hepatitis D virus; HPLC, high pressure liquid chromatography ; MEK, methyl ethyl ketone; MTBE, methyl tertiary butyl ether; NARTI or NRTI, nucleoside reverse transcriptase inhibitor; NtARTI or NtRTI, nucleotide analog reverse transcriptase inhibitor; NMR, nuclear magnetic resonance; PLM, polarization Light microscopy; RH, relative humidity; rcDNA, relaxed circular DNA; sAg, surface antigen; SFC, supercritical fluid chromatography; TGA, thermogravimetric analysis; THF, tetrahydrofuran; TLC, thin layer chromatography; XRPD, X-ray Powder diffraction; XRDP, X-ray diffraction pattern.

Ranges: Throughout this disclosure, various aspects of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have explicitly disclosed all possible subranges and individual numerical values within that range. For example, a description of a range such as from 1 to 6 should be considered to have explicitly disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as individual values within the range, eg, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. For example, a range of "about 0.1% to about 5%" or "about 0.1% to 5%" should be considered to include not only about 0.1% to about 5%, but also individual values within the indicated range (eg, 1%, 2%, 3%, and 4%) and subranges (eg, 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%). Unless otherwise indicated, the statement "about X to Y" has the same meaning as "about X to about Y". Likewise, the statement "about X, Y, or Z" has the same meaning as "about X, about Y, or about Z" unless stated otherwise. This applies regardless of the breadth of the scope.

Compounds and Synthesis

The present disclosure further provides methods of making the compounds of the present disclosure. The compounds of the present teachings can be prepared according to the procedures outlined herein from commercially available starting materials, compounds known in the literature, or by intermediates readily prepared using standard synthetic methods and procedures known to those skilled in the art. Standard synthetic methods and procedures for the preparation of organic molecules and functional group transformation and manipulation can be readily obtained from the relevant scientific literature or from standard textbooks in the field.

It should be appreciated that where typical or preferred process conditions (ie, reaction temperatures, times, molar ratios of reactants, solvents, pressures, etc.) are given, other process conditions may also be used unless otherwise specified. Optimal reaction conditions may vary with the particular reactants or solvents used, but such conditions can be determined by one skilled in the art by routine optimization procedures. Those skilled in the art of organic synthesis will recognize that the nature and order of the presented synthetic steps may be varied in order to optimize the formation of the compounds described herein.

The processes described herein can be monitored according to any suitable method known in the art. For example, by spectroscopic means, such as nuclear magnetic resonance spectroscopy (eg, ¹ H or ¹³ C), infrared spectroscopy, spectrophotometry (eg, UV-visible light), mass spectrometry, or by chromatography, such as high performance liquid Product formation was monitored by chromatography (HPLC), gas chromatography (GC), gel permeation chromatography (GPC) or thin layer chromatography (TLC).

The preparation of compounds can involve the protection and deprotection of various chemical groups. The need for protection and deprotection and the selection of appropriate protecting groups can be readily determined by those skilled in the art. The chemistry of protecting groups can be found, for example, in Greene, et al. , Protective Groups in Organic synthesis, 2d. Ed. (Wiley & Sons, 1991), which is hereby incorporated by reference in its entirety for all purposes .

The reactions or processes described herein can be carried out in suitable solvents readily selected by those skilled in the art of organic synthesis. Suitable solvents are typically substantially unreactive with the reactants, intermediates, and/or products at temperatures at which the reaction is carried out (ie, temperatures that can range from the solvent's freezing temperature to the solvent's boiling temperature). A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the specific reaction step, an appropriate solvent can be selected for the specific reaction step.

(X)， The present disclosure includes the preparation of ( R )-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxo-1,2-dihydroisoquinoline) Lin-4-yl)ethyl)-1-methylurea (also known as compound (X)), or its salts, solvates, prodrugs, isotopically-labeled derivatives, stereoisomers (such as, in non- In a limiting example, an enantiomer or any mixture thereof, such as, in a non-limiting example, a mixture of its enantiomers in any ratio), and/or a tautomer, and any mixture thereof method:

(X) ,

In certain embodiments, compounds of formula (X) , or salts, solvates, prodrugs, isotopically-labeled derivatives and/or tautomers thereof, can be synthesized according to the non-limiting synthetic scheme outlined in Scheme 1 preparation.

By Lewis acid catalyzed coupling with acetylacetone (using a cuprous (I) salt in a non-limiting example) in the presence of a base such as, but not limited to, an alkali metal alkoxide, the Commercially available bromide (A) is converted to the corresponding ketone (B) .

Ketones (B) can undergo acid-catalyzed cyclization in dichloromethane (DCM) to give isochromenone (C) , which can be reacted with formamide in basic solution to give isoquinolinone (D) . Alternatively, under acidic conditions in the presence of methanol, ketone (B) can undergo both acid-catalyzed esterification to yield ester (C') and acid-catalyzed cyclization to yield isochromene (C) , either Independently reacts with tris(D) under basic conditions to form isoquinolinones (D) .

In the presence of a tertiary base such as but not limited to triethylamine (TEA), Hünig base (diisopropylethylamine or DIPEA), pyridine, etc., in an inert solvent such as, but not limited to, acetonitrile, dichloromethane, etc., can be used Chlorinating agents, such as, but not limited to, phosphorus oxychloride, thionite chloride, and the like, convert the isoquinolinone (D) to the corresponding 1-chloroisoquinoline (E) .

The 1-chloroisoquinoline (E) can be converted to the corresponding 1-methoxyisoquinoline (F) using, for example, an alkali metal methoxide in methanol solution.

1-Methoxyisoquinoline (F) can, for example, be converted to sulfenamide (H) in a two-step process in the presence of a Lewis acid such as but not limited to a titanium (IV) alkoxide The following (F) is reacted with (G) to produce N-alkylenesulfinamides, which can be reduced to sulfenamides (H) using reducing agents such as, but not limited to, borohydrides.

The sulfinamide (H) can be made on the Alkylation at the sulfinamide nitrogen yields chiral intermediates (I) , which can be hydrolyzed to the corresponding isoquinolinones (J) using acids such as, but not limited to, hydrochloric acid, sulfuric acid, and the like .

Carbamate (K) can be prepared by catalyzing 3-cyano in the presence of a tertiary base such as, but not limited to, triethylamine (TEA), Hünig base (diisopropylethylamine or DIPEA), pyridine, and the like. It is prepared by reacting phenyl-4-fluoroaniline with phenyl chloroformate.

Compound (J) and compound (K) can be coupled in the presence of a tertiary amine base, such as, but not limited to, triethylamine (TEA), Hünig base (diisopropylethylamine or DIPEA), pyridine, etc., to generate Compound (X). In certain embodiments, (X) is isolated as free acid/base.

process 1

In certain embodiments, at least one compound of the present invention is a component of a pharmaceutical composition further comprising at least one pharmaceutically acceptable carrier.

Compounds contemplated by this invention may possess one or more stereocenters, and each stereocenter may independently exist in the ( R ) or ( S ) configuration. In certain embodiments, the compounds described herein exist in optically active or racemic forms. The compounds described herein include racemic, optically active, regioisomeric and stereoisomeric forms or combinations thereof having the therapeutically useful properties described herein. The preparation of optically active forms is accomplished in any suitable manner, including, by way of non-limiting example, resolution of racemic forms by recrystallization techniques, synthesis from optically active starting materials, chiral synthesis, or the use of chiral fixation The phases were separated by chromatography. In certain embodiments, mixtures of one or more isomers are used as therapeutic compounds described herein. In other embodiments, the compounds described herein contain one or more chiral centers. These compounds are prepared by any means, including stereoselective synthesis, enantioselective synthesis, and/or separation of mixtures of enantiomers and/or diastereomers. Resolution of compounds and isomers thereof can be accomplished by any method including, by way of non-limiting example, chemical methods, enzymatic methods, fractional crystallization, distillation, and chromatography.

In certain embodiments, compounds of the present disclosure exist as tautomers. All tautomers are included within the scope of the compounds described herein.

The compounds described herein also include isotopically labeled compounds in which one or more atoms are replaced with an atom having the same atomic number but an atomic mass or mass number different from that normally found in nature. Examples of isotopes suitable for inclusion in the ^compounds described herein include, but are not limited to,2H, ^3H , ^11C , ^13C , ^14C , ^36Cl , ^18F , ^123I , ^125I , ^13N , ^15N , ¹⁵ O, ¹⁷ O, ¹⁸ O, ³² P and ³⁵ S. In certain embodiments, isotopically labeled compounds are useful in drug and/or stromal tissue distribution studies. In other embodiments, substitution with heavier isotopes such as deuterium provides greater metabolic stability (eg, increased in vivo half-life or reduced dosage requirements). In yet other embodiments, substitution with positron emitting isotopes such ^as11C , ^18F , ^15O , ^and13N can be used in positron emission tomography (PET) studies to examine matrix receptor occupancy. Isotopically-labeled compounds can be prepared by any suitable method or by a process using an appropriate isotopic label in place of an otherwise employed unlabeled reagent.

In certain embodiments, the compounds described herein are labeled by other means, including, but not limited to, the use of chromophore or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

The compounds described herein, as well as other related compounds with various substituents, are used as described herein and/or, for example, as in Fieser &Fieser's Reagents for Organic Synthesis, Vol. 1-17 (John Wiley and Sons, 1991), Rodd's Chemistry of Carbon Compounds, Vol. 1-5 and Supplementals (Elsevier Science Publishers, 1989), Organic Reactions, Vol. 1-40 (John Wiley and Sons, 1991), Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989), March, Advanced Organic Chemistry 4th ^Ed ., (Wiley 1992), Carey & Sundberg, Advanced Organic Chemistry, 4th ^Ed ., Vols. A and B (Plenum 2000, 2001) and Green & Wuts, Protective Groups in Organic Synthesis 3rd Ed. , (Wiley 1999), all of which are incorporated by reference into the present disclosure, were synthesized using techniques and materials. The general methods for preparing the compounds described herein are modified by the use of appropriate reagents and conditions to incorporate the various moieties found in the formulae provided herein.

In all embodiments provided herein, examples of suitable optional substituents are not intended to limit the scope of the claimed disclosure. The compounds of the present disclosure can contain any substituent or combination of substituents provided herein.

Salt

The compounds described herein may form salts with acids or bases, and such salts are included in the present disclosure. The term "salt" includes addition salts of free acids or bases useful in the methods of the present disclosure. The term "pharmaceutically acceptable salt" refers to a salt having a toxicity profile within a range useful in pharmaceutical applications. In certain embodiments, the salt is a pharmaceutically acceptable salt. However, pharmaceutically unacceptable salts may possess properties such as high crystallinity, which have utility in the practice of the present invention, such as, for example, utility in the synthesis, purification or formulation of compounds useful in the methods of the present disclosure sex.

Suitable pharmaceutically acceptable acid addition salts can be prepared from inorganic or organic acids. Examples of inorganic acids include sulfate, hydrogen sulfate, hydrochloric acid, hydrobromic acid, hydroiodic acid, nitric acid, carbonic acid, sulfuric acid, and phosphoric acid (including hydrogen phosphate and dihydrogen phosphate). Suitable organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic acids, examples of which include formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, glucose Sugar acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, 4 -Hydroxybenzoic acid, phenylacetic acid, mandelic acid, pamoic acid (or pamoic acid), methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, pantothenic acid, sulfanilic acid, 2-hydroxyethanesulfonic acid, trifluoromethanesulfonic acid, p-toluenesulfonic acid, cyclohexylaminosulfonic acid, stearic acid, alginic acid, beta-hydroxybutyric acid, salicylic acid, galactaric acid, galacturonic acid, glycerophosphonic acid, and saccharin (e.g., saccharin salts ( saccharinate), saccharate). A salt may consist of 1 molar equivalent, 1 or more than 1 molar equivalent of an acid or base moiety relative to any compound of the present disclosure.

Suitable pharmaceutically acceptable base addition salts of the compounds of the present disclosure include, for example, ammonium and metal salts, including alkali metal, alkaline earth metal and transition metal salts such as, for example, calcium, magnesium, potassium, sodium and zinc salts. Pharmaceutically acceptable base addition salts also include compounds made from basic amines such as, for example, N , N' -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (or N -methylglucamine) and organic salts prepared from procaine. All of these salts can be prepared from the corresponding compounds by, for example, reacting a suitable acid or base with the compound.

combination therapy

In one aspect, the compounds of the present invention are used in the methods of the present invention in combination with one or more additional agents for the treatment of HBV and/or HDV infection. These additional agents may include compounds or compositions identified herein, or compounds (eg, commercially available compounds) known to be useful in the treatment, prevention, or alleviation of symptoms of HBV and/or HDV infection.

Non-limiting examples of one or more additional agents for treating HBV and/or HDV infection include: (a) reverse transcriptase inhibitors; (b) capsid inhibitors; (c) cccDNA formation inhibitors; (d) ) RNA destabilizers; (e) oligonucleotides targeting the HBV genome; (f) immunostimulants such as checkpoint inhibitors (eg, PD-L1 inhibitors); (g) targeting HBV gene transcription and (h) a therapeutic vaccine.

(a) reverse transcriptase inhibitor

In certain embodiments, the reverse transcriptase inhibitor is a reverse transcriptase inhibitor (NARTI or NRTI). In other embodiments, the reverse transcriptase inhibitor is a nucleotide analog reverse transcriptase inhibitor (NtARTI or NtRTI).

Reported reverse transcriptase inhibitors include, but are not limited to, entecavir, clevudine, telbivudine, lamivudine, adefovir, and tenofovir tenofovir, tenofovir disoproxil, tenofovir alafenamide, adefovir dipovoxil, (1 R ,2 R ,3 R , 5R )-3-(6-amino-9H-9- purinyl )-2-fluoro-5-(hydroxymethyl)-4-methylenecyclopentan-1-ol (described in U.S. Patent 8,816,074, incorporated herein by reference in its entirety), emtricitabine, abacavir, elvucitabine, ganciclovir, lobcavir, famciclovir, penciclovir and Amdoxovir.

Reported reverse transcriptase inhibitors further include, but are not limited to, entecavir, lamivudine, and ( 1R , 2R , 3R , 5R )-3-(6-amino-9H-9- purinyl )- 2-Fluoro-5-(hydroxymethyl)-4-methylenecyclopentan-1-ol.

Reported reverse transcriptase inhibitors further include, but are not limited to, covalently bound aminophosphonate or phosphatamide moieties of the reverse transcriptase inhibitors mentioned above, or as described, for example, in U.S. Patent No. 8,816,074, U.S. Patent Application Publication No. US 2011/0245484 A1 and US 2008/0286230 A1, all of which are incorporated herein by reference in their entirety.

Reported reverse transcriptase inhibitors further include, but are not limited to, nucleotide analogs that include a phosphoramidate moiety, such as, for example, (((( 1R , 3R , 4R , 5R )-3-(6-amine yl-9H-purin-9-yl)-4-fluoro-5-hydroxy-2-methylenecyclopentyl)methoxy)methyl(phenoxy)phosphoryl)-( D or L) -Alanine ester and ((((1 R ,2 R ,3 R ,4 R )-3-fluoro-2-hydroxy-5-methylene-4-(6-oxy-1,6-dihydro -9H-purin-9-yl)cyclopentyl)methoxy)methyl(phenoxy)phosphoryl)-( D or L)-alanine ester. Also included are its individual diastereomers, including, for example, (( R )-((( 1R , 3R , 4R , 5R )-3-(6-amino- 9H -purine-9 -yl)-4-fluoro-5-hydroxy-2-methylenecyclopentyl)methoxy)methyl(phenoxy)phosphoryl)-(D or L)-alanine ester and (( S )-(((1 R ,3 R ,4 R ,5 R )-3-(6-amino-9 H -purin-9-yl)-4-fluoro-5-hydroxy-2-methylene ring Amyl)methoxy)methyl(phenoxy)phosphoryl)-(D or L)-alanine ester.

Reported reverse transcriptase inhibitors further include, but are not limited to, compounds that include a phosphamide moiety, such as, for example, tenofovir alafenamide, and those described in US Patent Application Publication No. US 2008/0286230 A1, by This reference is incorporated herein in its entirety. Methods for preparing stereoselective aminophosphate or phosphamidamide containing actives are described, for example, in US Patent No. 8,816,074 and US Patent Application Publication Nos. US 2011/0245484 A1 and US 2008/0286230 A1, all of which It is incorporated herein by reference in its entirety.

(b) capsid inhibitor

As described herein, the term "capsid inhibitor" includes compounds capable of directly or indirectly inhibiting the expression and/or function of capsid proteins. For example, capsid inhibitors can include, but are not limited to, inhibiting capsid assembly, inducing non-capsid polymer formation, promoting excess capsid assembly or misdirection of capsid assembly, affecting capsid stability, and/or inhibiting RNA (pgRNA) coating any compound that is shelled. Capsid inhibitors are also included in the downstream event(s) of the replication process (e.g., viral DNA synthesis, translocation of loose-loop DNA (rcDNA) to the nucleus, covalently closed circular DNA (cccDNA) formation, viral maturation, budding and/or release, etc.) that inhibit capsid function. For example, in certain embodiments, the inhibitor detectably inhibits the expression level or biological activity of the capsid protein, eg, as measured using the assays described herein. In certain embodiments, the inhibitor inhibits the levels of rcDNA and downstream products of the viral life cycle by at least 5%, at least 10%, at least 20%, at least 50%, at least 75%, or at least 90%.

Reported capsid inhibitors include, but are not limited to, compounds described in International Patent Application Publication Nos. WO 2013006394, WO 2014106019, and WO2014089296, all of which are incorporated herein by reference in their entirety.

Reported capsid inhibitors also include but are not limited to the following compounds and their pharmaceutically acceptable salts and/or solvates: Bay-41-4109 (see International Patent Application Publication No. WO 2013144129), AT-61 (see International Patent Application Publication No. WO 2013144129) Patent Application Publication No. WO 1998033501; and King et al., 1998, Antimicrob. Agents Chemother. 42(12):3179-3186), DVR-01 and DVR-23 (see International Patent Application Publication No. WO 2013006394; and Campagna et al. al., 2013, J. Virol. 87(12):6931, which is hereby incorporated by reference in its entirety.

Additionally, reported capsid inhibitors include, but are not limited to, those described in US Patent Application Publication Nos. US 2015/0225355, US 2015/0132258, US 2016/0083383, US 2016/0052921 and International Patent Application Publication Nos. WO 2013096744, WO 2014165128, WO 2014033170、WO 2014033167、WO 2014033176、WO 2014131847、WO 2014161888、WO 2014184350、WO 2014184365、WO 2015059212、WO 2015011281、WO 2015118057、WO 2015109130、WO 2015073774、WO 2015180631、WO 2015138895、WO 2016089990、WO 2017015451、WO 2016183266、 Those generally and specifically described in WO 2017011552, WO 2017048950, WO2017048954, WO 2017048962, WO 2017064156, and are hereby incorporated by reference in their entirety.

(c) cccDNA formation inhibitor

Covalently closed circular DNA (cccDNA) is produced in the nucleus of viral rcDNA and serves as the transcription template for viral mRNA. As described herein, the term "inhibitor of cccDNA formation" includes compounds capable of directly or indirectly inhibiting the formation and/or stability of cccDNA. For example, inhibitors of cccDNA formation can include, but are not limited to, any compound that inhibits capsid disassembly, entry of rcDNA into the nucleus, and/or conversion of rcDNA to cccDNA. For example, in certain embodiments, the inhibitor inhibits the formation and/or stability of cccDNA, eg, as measured using the assays described herein. In certain embodiments, the inhibitor inhibits the formation and/or stability of cccDNA by at least 5%, at least 10%, at least 20%, at least 50%, at least 75%, or at least 90%.

Reported inhibitors of cccDNA formation include, but are not limited to, compounds described in International Patent Application Publication No. WO 2013130703, which is incorporated herein by reference in its entirety.

In addition, reported inhibitors of cccDNA formation include, but are not limited to, those generally and specifically described in US Patent Application Publication No. US 2015/0038515 Al, which is hereby incorporated by reference in its entirety.

(d) RNA destabilizer

As used herein, the term "RNA destabilizer" refers to a molecule or a salt or solvate thereof that reduces the total amount of HBV RNA in mammalian cell culture or in a living human subject. In a non-limiting example, the RNA destabilizer reduces the amount of RNA transcript(s) encoding one or more of the following HBV proteins: surface antigen, core protein, RNA polymerase, and e-antigen. In certain embodiments, the RNA destabilizer reduces the total amount of HBV RNA in mammalian cell culture or in a live human subject by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or at least 90%.

Reported RNA destabilizers include compounds described in US Pat. No. 8,921,381, as well as compounds described in US Patent Application Publication Nos. US 2015/0087659 and US 2013/0303552, all of which are incorporated herein by reference in their entirety.

In addition, the reported RNA de -stabilizer includes but is not limited to but not limited to the open number of WO 2015113990, WO 2015173164, US 2016/0122344, WO 2016107832, WO 2016023877, WO 2016128335, WO 2016176555, WO 2017901346, wo 201701346, wo27017013466 Those generally and specifically described in WO 2017016960, WO 2017017042, WO 2017017043, WO 2017102648, WO 2017108630, WO 2017114812, WO 2017140821, WO 2018085619, and are hereby incorporated by reference in their entirety.

(e) target HBV genomic oligonucleotides

Reported oligonucleotides targeting the HBV genome include, but are not limited to, Arrowhead-ARC-520 (see US Pat. No. 8,809,293; and Wooddell et al., 2013, Molecular Therapy 21(5):973-985, all by is incorporated herein by reference in its entirety).

In certain embodiments, oligonucleotides can be designed to target one or more genes and/or transcripts of the HBV genome. Oligonucleotides targeting the HBV genome also include, but are not limited to, isolated double-stranded siRNA molecules, each siRNA molecule comprising a sense strand and an antisense strand that hybridizes to the sense strand. In certain embodiments, the siRNA targets one or more genes and/or transcripts of the HBV genome.

(f) immune stimulants

Checkpoint inhibitors

As described herein, the term "checkpoint inhibitor" includes any compound capable of inhibiting immune checkpoint molecules that act as immune system modulators (eg, stimulate or inhibit immune system activity). For example, some checkpoint inhibitors block inhibitory checkpoint molecules, which stimulate immune system function, such as stimulating T cell activity against cancer cells. A non-limiting example of a checkpoint inhibitor is a PD-L1 inhibitor.

As described herein, the term "PD-L1 inhibitor" includes any compound capable of directly or indirectly inhibiting the expression and/or function of programmed death ligand 1 (PD-L1) protein. PD-L1, also known as cluster of differentiation 274 (CD274) or B7 homolog 1 (B7-H1), is important in suppressing the adaptive arm of the immune system during pregnancy, tissue allotransplantation, autoimmune disease and hepatitis Acting type 1 transmembrane protein. PD-L1 binds to its receptor, the inhibitory checkpoint molecule PD-1 (found on activated T cells, B cells and myeloid cells) in order to regulate the activation or inhibition of the adaptive arm of the immune system. In certain embodiments, the PD-L1 inhibitor inhibits the expression and/or function of PD-L1 by at least 5%, at least 10%, at least 20%, at least 50%, at least 75%, or at least 90%.

Reported PD-L1 inhibitors include, but are not limited to, compounds described in one of the following patent application publications: US 2018/0057455; US 2018/0057486; WO 2017/106634; WO 2018/026971; WO 2018/045142; WO 2018/118848; WO 2018/119221; WO 2018/119236; WO 2018/119266; WO 2018/119286; .

(g) target HBV gene transcript GalNAc-siRNA Conjugate

"GalNAc" is an abbreviation for N-acetylgalactosamine, and "siRNA" is an abbreviation for small interfering RNA. In GalNAc-siRNA conjugates useful in the practice of the present invention, siRNA targeting the HBV gene transcript is covalently bound to GalNAc. While not wishing to be bound by theory, it is believed that GalNAc binds to asialoglycoprotein receptors on hepatocytes, thereby facilitating siRNA targeting of HBV-infected hepatocytes. siRNA enters infected hepatocytes and stimulates the destruction of HBV gene transcripts by the phenomenon of RNA interference.

Examples of GalNAc-siRNA conjugates useful in the practice of this aspect of the invention are in published International Application PCT/CA2017/050447 (PCT Application Publication No. WO/2017/177326 published October 19, 2017) set forth, which is incorporated herein by reference in its entirety.

(h) therapeutic vaccine

In certain embodiments, administration of a therapeutic vaccine is useful in the practice of the present disclosure for treating a viral disease in a subject. In certain embodiments, the viral disease is hepatitis virus. In certain embodiments, the hepatitis virus is at least one selected from the group consisting of hepatitis B virus (HBV) and hepatitis D virus (HDV). In certain embodiments, the subject is a human.

For example, suitable methods such as, for example, the Sigmoid- _Emax equation (Holford & Scheiner, 1981, Clin. Pharmacokinet. 6:429-453), the Loewe additivity equation (Loewe & Muischnek, 1926, Arch. Exp. Pathol) can be used Pharmacol. 114:313-326) and the median effect equation (Chou & Talalay, 1984, Adv. Enzyme Regul. 22:27-55) calculated synergistic effects. Each of the equations mentioned elsewhere in this article can be applied to experimental data to generate corresponding curves to help assess the effects of drug combinations. The corresponding curves associated with the equations mentioned elsewhere in this document are the concentration-effect curve, the isobologram curve and the combined exponential curve, respectively.

method

In one aspect, the present disclosure provides a method of treating or preventing hepatitis virus infection in a subject. In certain embodiments, the infection comprises hepatitis B virus (HBV) infection. In other embodiments, the methods comprise administering to a subject in need thereof a therapeutically effective amount of at least one compound of the present invention. In yet other embodiments, at least one compound is administered to the subject in a pharmaceutically acceptable composition. In yet other embodiments, the subject is further administered at least one additional agent for the treatment of hepatitis infection. In yet other embodiments, the at least one additional agent comprises at least one selected from the group consisting of: a reverse transcriptase inhibitor; a capsid inhibitor; an inhibitor of cccDNA formation; an RNA destabilizer; acid; immunostimulants, such as checkpoint inhibitors (eg, PD-L1 inhibitors); GalNAc-siRNA conjugates targeting HBV gene transcripts; and therapeutic vaccines. In yet other embodiments, the subject is co-administered at least one compound and at least one additional agent. In yet other embodiments, at least one compound and at least one additional agent are co-formulated.

The present invention further provides methods of directly or indirectly inhibiting the expression and/or function of viral capsid proteins in a subject. In certain embodiments, the methods comprise administering to a subject in need thereof a therapeutically effective amount of at least one compound of the present invention. In other embodiments, at least one compound is administered to the subject in a pharmaceutically acceptable composition. In yet other embodiments, the subject is further administered at least one additional agent for treating HBV infection. In yet other embodiments, the at least one additional agent comprises at least one selected from the group consisting of: a reverse transcriptase inhibitor; a capsid inhibitor; an inhibitor of cccDNA formation; an RNA destabilizer; acid; immunostimulants - such as checkpoint inhibitors (eg, PD-L1 inhibitors); GalNAc-siRNA conjugates targeting HBV gene transcripts; and therapeutic vaccines. In yet other embodiments, the subject is co-administered at least one compound and at least one additional agent. In yet other embodiments, at least one compound and at least one additional agent are co-formulated.

In certain embodiments, the subject is a mammal. In other embodiments, the mammal is a human.

Pharmaceutical compositions and preparations

The present invention provides pharmaceutical compositions comprising at least one compound of the present invention, or a salt or solvate thereof, for use in practicing the methods of the present invention. Such a pharmaceutical composition may consist of at least one compound of the present invention or a salt or solvate thereof in a form suitable for administration to a subject, or the pharmaceutical composition may comprise at least one compound of the present invention or a salt or solvate thereof and one or Various pharmaceutically acceptable carriers, one or more additional ingredients, or any combination of these. As is well known in the art, at least one compound of the present invention may be present in a pharmaceutical composition in the form of a physiologically acceptable salt, eg, in combination with a physiologically acceptable cation or anion.

In certain embodiments, pharmaceutical compositions for practicing the methods of the present invention can be administered to deliver a dose of between 1 ng/kg/day and 100 mg/kg/day. In other embodiments, the pharmaceutical compositions used to practice the present invention can be administered to deliver a dose of between 1 ng/kg/day and 1,000 mg/kg/day.

The relative amounts of active ingredients, pharmaceutically acceptable carriers and any additional ingredients in the pharmaceutical compositions of the present invention will vary depending on the identity, size and condition of the subject being treated, and further Depends on the route by which the composition is administered. For example, the composition may include between 0.1% and 100% (w/w) active ingredient.

The pharmaceutical composition used in the method of the present invention can be appropriately developed for nasal, inhalation, oral, rectal, vaginal, pleural, peritoneal, parenteral, topical, transdermal, pulmonary, intranasal, buccal, ophthalmic Intradural, intrathecal, intravenous, or another route of administration. The compositions used in the methods of the present invention can be administered directly to the brain, brain stem, or any other part of the central nervous system of a mammal or avian. Other contemplated formulations include projected nanoparticles, microspheres, liposomal formulations, coated particles, polymer conjugates, resealed red blood cells containing active ingredients, and immunology-based formulations.

In certain embodiments, the compositions of the present invention are part of a pharmaceutical matrix that allows for the processing of insoluble materials and their improved bioavailability, the development of controlled or sustained release products, and the creation of homogeneous compositions. For example, hot melt extrusion, solid solutions, solid dispersions, size reduction techniques, molecular complexes (eg, cyclodextrins, etc.), microparticles, and granule and formulation coating methods can be used to prepare drug matrices. Amorphous or crystalline phases can be used in this process.

The route(s) of administration will be apparent to the skilled artisan and will depend on many factors, including the type and severity of the disease being treated, the type and age of the veterinary or human patient being treated, and the like.

The formulations of the pharmaceutical compositions described herein can be prepared by any method known or hereafter developed in the art of pharmacology and pharmacy. Generally, such methods of preparation include the steps of bringing into association the active ingredient with a carrier or one or more other accessory ingredients, and then, if necessary or desired, shaping or packaging the product in the desired single- or multi-dose unit.

As used herein, a "unit dose" is a discrete quantity of a pharmaceutical composition containing a predetermined quantity of an active ingredient. The amount of active ingredient is generally equal to the dose of active ingredient to be administered to the subject or a convenient fraction of such a dose, such as, for example, one half or one-third of such a dose. The unit dosage form can be a single daily dose or one of multiple daily doses (eg, about 1-4 or more times per day). When multiple daily doses are used, the unit dosage form for each administration may be the same or different.

Although the descriptions of pharmaceutical compositions provided herein are primarily directed to pharmaceutical compositions suitable for ethical administration to humans, the skilled artisan will understand that such compositions are generally suitable for administration to various animals. Modifications of pharmaceutical compositions suitable for administration to humans in order to make compositions suitable for administration to a variety of animals are well known and can be devised and carried out by a veterinary pharmacologist of ordinary skill only by ordinary experimentation. Subjects to which the pharmaceutical compositions of the present invention are expected to be administered include, but are not limited to, humans and other primates, mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, and dogs .

In certain embodiments, the compositions of the present invention are formulated using one or more pharmaceutically acceptable excipients or carriers. In certain embodiments, the pharmaceutical compositions of the present invention include a therapeutically effective amount of at least one compound of the present invention and a pharmaceutically acceptable carrier. Useful pharmaceutically acceptable carriers include, but are not limited to, glycerol, water, saline, ethanol, recombinant human albumin (eg, Recombumin®), soluble gels (eg, Gelofusine®), and other pharmaceutically acceptable salts Solutions such as salts of phosphates and organic acids. Examples of these and other pharmaceutically acceptable carriers are described in Remington's Pharmaceutical Sciences (1991, Mack Publication Co., New Jersey).

The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), recombinant human albumin, soluble gelatin, suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of coatings such as lecithin, by the maintenance of the desired particle size in the presence of dispersions, and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, isotonic agents such as sugars, sodium chloride or polyols such as mannitol and sorbitol are included in the composition. Prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate or gelatin.

The formulations may be used in admixture with conventional excipients, ie pharmaceutically acceptable, suitable for oral, parenteral, nasal, inhalation, intravenous, subcutaneous, transdermal enteral or any other suitable mode of administration known in the art of organic or inorganic carrier substances. The pharmaceutical preparations can be sterilized and, if desired, mixed with adjuvants such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing the osmotic pressure buffer, coloring agents, flavoring agents and/or Aroma-imparting substances, etc. are mixed. They can also be combined with other active agents, such as other analgesics, anxiolytics or hypnotics, if desired. As used herein, "additional ingredients" include, but are not limited to, one or more ingredients that can be used as a pharmaceutical carrier.

The compositions of the present invention may include from about 0.005% to 2.0% of a preservative by total weight of the composition. Preservatives are used to prevent spoilage in the event of exposure to contaminants in the environment. Examples of preservatives useful according to the present invention include, but are not limited to, those selected from the group consisting of benzyl alcohol, sorbic acid, p-hydroxybenzoic acid, imidurea, and combinations thereof. One such preservative is a combination of about 0.5%-2.0% benzyl alcohol and 0.05%-0.5% sorbic acid.

The composition may include antioxidants and chelating agents that inhibit the degradation of the compounds. For certain compounds, the antioxidants are BHT, BHA, alpha-tocopherol, and ascorbic acid, with an exemplary range of about 0.01% to 0.3% by weight, based on the total weight of the composition, or a BHT range of 0.03% by weight % to 0.1%. The chelating agent may be present in an amount of 0.01% to 0.5% by weight, based on the total weight of the composition. Exemplary chelating agents include in the range of about 0.01% to 0.20% by weight or in the range of 0.02% to 0.10% by weight ethylenediaminetetraacetate (eg, ethylenediaminetetraacetate), based on the total weight of the composition disodium acetate) and citric acid. Chelating agents can be used to chelate metal ions in the composition, which can be detrimental to the shelf life of the formulation. For certain compounds, although BHT and disodium EDTA are exemplary antioxidants and chelating agents, respectively, other suitable and equivalent antioxidants and chelating agents can be substituted as known to those skilled in the art.

Liquid suspensions can be prepared using conventional methods to suspend the active ingredient in aqueous or oily vehicles. Aqueous vehicles include, for example, water and isotonic saline. Oily vehicles include, for example, almond oil, oily esters, ethanol, vegetable oils such as peanut, olive, sesame or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin. Liquid suspensions may further include one or more additional ingredients including, but not limited to, suspending, dispersing or wetting agents, emulsifying agents, emulsifying agents, preservatives, buffers, salts, flavoring, coloring and sweetening agents . The oily suspensions may further comprise thickening agents. Known suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fats, sodium alginate, polyvinylpyrrolidone, tragacanth, acacia, and cellulose derivatives such as sodium carboxymethylcellulose, methyl cellulose, hydroxypropyl methylcellulose. Known dispersing or wetting agents include, but are not limited to, naturally occurring phospholipids such as lecithin, alkylene oxides with fatty acids, with long-chain aliphatic alcohols, with partial esters derived from fatty acids and hexitols, or with fatty acids and hexitols. Partial esters of anhydrosugar alcohols (for example, polyoxyethylene stearate, heptaethyleneoxycetyl alcohol, polyoxyethylene sorbitan monooleate, and polyoxyethylene sorbitan monooleate, respectively) ) condensation product. Known emulsifiers include, but are not limited to, lecithin, acacia, and ionic or nonionic surfactants. Known preservatives include, but are not limited to, methylparaben, ethylparaben, or n-propylparaben, ascorbic acid, and sorbic acid. Known sweeteners include, for example, glycerol, propylene glycol, sorbitol, sucrose, and saccharin.

Liquid solutions of the active ingredient in aqueous or oily solvents can be prepared in substantially the same manner as liquid suspensions, with the main difference being that the active ingredient is dissolved rather than suspended in the solvent. As used herein, an "oily" liquid is a liquid that includes carbon-containing liquid molecules and exhibits less polarity than water. Liquid solutions of the pharmaceutical compositions of the present invention may contain each of the components described with respect to liquid suspensions, it being understood that suspending agents do not necessarily aid dissolution of the active ingredient in the solvent. Aqueous solvents include, for example, water and isotonic saline. Oily solvents include, for example, almond oil, oily esters, ethanol, vegetable oils such as peanut oil, olive oil, sesame oil or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin.

Powder and granular formulations of the pharmaceutical formulations of the present invention can be prepared using known methods. Such formulations can be administered directly to a subject, for example, to form lozenges, fill capsules, or to prepare aqueous or oily suspensions or solutions by adding thereto an aqueous or oily vehicle. Each of these formulations may further include one or more of dispersing or wetting agents, suspending agents, ionic and nonionic surfactants, and preservatives. Additional excipients such as fillers and sweetening, flavoring or coloring agents may also be included in these formulations.

The pharmaceutical compositions of the present invention can also be prepared, packaged, or sold in the form of oil-in-water emulsions or water-in-oil emulsions. The oily phase can be a vegetable oil such as olive or peanut oil, a mineral oil such as liquid paraffin, or a combination of these. Such compositions may further comprise one or more emulsifiers such as naturally occurring gums such as acacia or tragacanth; naturally occurring phospholipids such as soy or lecithin; esters or partials derived from combinations of fatty acids and hexitol anhydrides Esters, such as sorbitan monooleate, and the condensation products of such partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. These emulsions may also contain additional ingredients including, for example, sweetening or flavoring agents.

Methods of impregnating or coating materials with chemical compositions are known in the art and include, but are not limited to, methods of depositing or binding chemical compositions to surfaces, in synthetic materials (ie, using, for example, physiologically acceptable methods of incorporating chemical compositions into the structure of materials during degraded materials) and methods of absorbing aqueous or oily solutions or suspensions into absorbent materials, followed by drying or no drying. Methods of mixing the components include physical milling, use of pellets in solid and suspension formulations, and mixing in transdermal patches known to those skilled in the art.

cast / dosing

The dosing regimen may affect the composition of the effective amount. The therapeutic formulation can be administered to the patient before or after the onset of disease or symptoms. In addition, several divided and staggered doses may be administered daily or sequentially, or the doses may be infused continuously, or they may be bolus injections. Furthermore, the dosage of the therapeutic agent may be proportionally increased or decreased as indicated by the exigencies of the therapeutic or prophylactic situation.

The compositions of the present invention can be administered to a patient, eg, a mammal, eg, a human, using known procedures, at doses and for periods of time effective to treat the disease or condition contemplated herein. The effective amount of a therapeutic compound necessary to achieve a therapeutic effect may vary depending on factors such as the activity of the particular compound employed; the time of administration; the rate of excretion of the compound; the duration of treatment; other drugs used in combination with the compound , compound or material; state of the disease or symptom, age, sex, weight, condition, general health and past medical history of the patient being treated, and similar factors well known in the medical arts. Dosage regimens can be adjusted to provide the best therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced, as indicated by the exigencies of the therapeutic situation. A non-limiting example of an effective dosage range of a therapeutic compound of the present invention is about 0.01 mg/kg to 100 mg/kg body weight per day. One of ordinary skill in the art would be able to study relevant factors and determine an effective amount of a therapeutic compound without undue experimentation.

The compound may be administered to the animal frequently several times a day, or may be administered less frequently, such as once a day, once a week, once every two weeks, once a month, or even less frequently, such as every few Once a month, or even once a year or less. It will be appreciated that, in non-limiting examples, the amount of compound to be administered per day may be administered once every day, every other day, every 2 days, every 3 days, every 4 days, or every 5 days. For example, administered every other day, a daily dose of 5 mg may be started on Monday, the first subsequent daily dose of 5 mg administered on Wednesday, and the second subsequent daily dose of 5 mg administered on Friday ,and many more. The frequency of dosing will be apparent to the skilled artisan and will depend on many factors such as, but not limited to, the type and severity of the disease being treated and the type and age of the animal.

The actual dosage level of the active ingredient in the pharmaceutical compositions of the present invention can be varied in order to obtain an amount of active ingredient effective to achieve the desired therapeutic response for a particular patient, composition and mode of administration, without being toxic to the patient.

A physician of ordinary skill in the art, such as a physician or veterinarian, can readily determine and prescribe the desired effective amount of the pharmaceutical composition. For example, a physician or veterinarian may start doses of the compounds of the invention used in pharmaceutical compositions at levels lower than required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

In particular embodiments, it is especially advantageous to formulate the compounds in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suitable as unitary dosages for the patients to be treated; each unit contains a predetermined quantity of the therapeutic compound calculated to be in association with the required pharmaceutical vehicle. produce the desired therapeutic effect. The dosage unit form of the present invention is determined by and is directly dependent on (a) the unique characteristics of the therapeutic compound and the particular therapeutic effect to be achieved, and (b) the time when such therapeutic compound is formulated/formulated for the treatment of a disease or condition in a patient inherent limitations in the field.

In certain embodiments, the compositions of the present invention are administered to a patient at a dose in the range of 1-5 or more times per day. In other embodiments, the compositions of the present invention are administered to a patient in a dosage range that includes, but is not limited to, once daily, once every two days, once every three days to once a week, and once every two weeks. It will be apparent to those skilled in the art that the administration of the various compositions of the present invention will vary from subject to subject, depending on a number of factors, including but not limited to age, disease or symptom being treated, gender, general health and other factors. Accordingly, the present invention should not be construed as limited to any particular dosage regimen, and the precise dosage and composition to be administered to any patient will be determined by the attending physician taking into account all other factors of the patient.

The compounds of the invention for administration may be in the following ranges: about 1 μg to about 7,500 mg, about 20 μg to about 7,000 mg, about 40 μg to about 6,500 mg, about 80 μg to about 6,000 mg, about 100 μg to about 5,500 mg, about 200 µg to about 5,000 mg, about 400 µg to about 4,000 mg, about 800 µg to about 3,000 mg, about 1 mg to about 2,500 mg, about 2 mg to about 2,000 mg, about 5 mg to about 1,000 mg, about 10 mg to about 750 mg, about 20 mg to about 600 mg, about 30 mg to about 500 mg, about 40 mg to about 400 mg, about 50 mg to about 300 mg, about 60 mg to about 250 mg , about 70 mg to about 200 mg, about 80 mg to about 150 mg, and any full and partial increments therebetween.

In some embodiments, the dose of a compound of the invention is between about 0.5 μg and about 5,000 mg . In some embodiments, the doses of the compounds of the invention used in the compositions described herein are less than about 5,000 mg, or less than about 4,000 mg, or less than about 3,000 mg, or less than about 2,000 mg, or less than about 1,000 mg, or Less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 200 mg, or less than about 50 mg. Similarly, in some embodiments, the dosage of the second compound as described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 400 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 40 mg, or less than about 30 mg, or less than about 25 mg, or less than about 20 mg, or less than about 15 mg , or less than about 10 mg, or less than about 5 mg, or less than about 2 mg, or less than about 1 mg, or less than about 0.5 mg, and any full and partial increments thereof.

In certain embodiments, the present invention relates to a packaged pharmaceutical composition comprising a container containing a therapeutically effective amount of a compound of the present invention, alone or in combination with a second drug; Instructions for one or more symptoms of a disease or condition.

The term "container" includes any receptacle for holding a pharmaceutical composition or for managing stability or water absorption. For example, in certain embodiments, the container is a package containing a pharmaceutical composition, such as liquids (solutions and suspensions), semi-solids, lyophilized solids, solutions and powders, or lyophilized formulations, present in dual compartments. In other embodiments, the container is not a package containing the pharmaceutical composition, ie, the container is a receptacle, such as a box or vial containing a packaged or unpackaged pharmaceutical composition and instructions for use of the pharmaceutical composition. Furthermore, packaging techniques are well known in the art. It will be appreciated that instructions for use of the pharmaceutical composition may be included on the package containing the pharmaceutical composition, and thus, the instructions form an increased functional relationship with the packaged product. It should be understood, however, that the instructions may contain information regarding the ability of the compound to perform its intended function, such as treating, preventing or alleviating a disease or symptom in a patient.

dosing

Routes of administration for any of the compositions of the present invention include inhalation, oral, nasal, rectal, parenteral, sublingual, transdermal, transmucosal (eg, sublingual, lingual, (trans) buccal, (trans) urethral, vaginal ( For example, transvaginal and perivaginal), nasal (intra) and ((trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastric, intrathecal, epidural, intrapleural, intraperitoneal, subcutaneous, intramuscular , intradermal, intraarterial, intravenous, intrabronchial, inhalation and topical administration.

Suitable compositions and dosage forms include, for example, troches, capsules, caplets, pills, softgels, lozenges, emulsions, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels Dosages, powders, pellets, magma, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, Dry powder or aerosol formulations for inhalation, compositions and formulations for intravesical administration, etc. It is to be understood that the formulations and compositions useful in the present invention are not limited to the specific formulations and compositions described herein.

oral administration

For oral administration, lozenges, lozenges, liquids, drops, capsules, caplets and softgels are particularly suitable. Other formulations suitable for oral administration include, but are not limited to, powder or granular formulations, aqueous or oily suspensions, aqueous or oily solutions, pastes, gels, toothpastes, mouthwashes, coatings, mouth rinses or emulsions. Compositions intended for oral use may be prepared according to any method known in the art, and such compositions may contain a drug selected from the group consisting of inert, nontoxic, generally regarded as safe (GRAS) pharmaceuticals suitable for use in the manufacture of lozenges One or more agents on excipients. Such excipients include, for example, inert diluents such as lactose; granulating and disintegrating agents such as corn starch; binding agents such as starch; and lubricants such as magnesium stearate.

Tablets may be uncoated or may be coated using known methods to achieve delayed disintegration in the gastrointestinal tract of a subject, thereby providing sustained release and absorption of the active ingredient. For example, a tablet may be coated with a material such as glyceryl monostearate or glyceryl distearate. By way of further example, lozenges may be coated to form osmotic controlled release tablets using the methods described in US Patent Nos. 4,256,108; 4,160,452; and 4,265,874. Lozenges may further contain sweetening, flavoring, coloring, preservative, or some combination of these to provide a pharmaceutically elegant and palatable preparation. Hard capsules containing the active ingredient can be prepared using physiologically degradable compositions such as gelatin. Capsules include the active ingredient and may further include additional ingredients including, for example, inert solid diluents such as calcium carbonate, calcium phosphate or kaolin.

Hard capsules containing the active ingredient can be prepared using physiologically degradable compositions such as gelatin. Such hard capsules contain the active ingredient and may further contain additional ingredients including, for example, inert solid diluents such as calcium carbonate, calcium phosphate or kaolin.

Soft capsules containing the active ingredient can be prepared using a physiologically degradable composition such as gelatin from animal collagen or hypromellose (a modified form of cellulose) using gelatin, water, and a plasticizer such as sorbitan Manufactured from an optional mixture of sugar alcohols or glycerol. Such soft capsules contain the active ingredient in admixture with an aqueous or oily vehicle, such as peanut oil, liquid paraffin, or olive oil.

For oral administration, the compounds of the present invention may be in the form of lozenges or capsules prepared by conventional methods with pharmaceutically acceptable excipients such as binders; fillers; lubricants; disintegrants; or wetting agents . If desired, suitable methods and coating materials can be used such as those available from Colorcon, West Point, Pa. (eg, OPADRY® Type OY, Type OYC, Organic Enteric OY-P, Aqueous Enteric OY- Form A, OY-PM and OPADRY® White, 32K18400) with the OPADRY® film coating system to coat tablets. It should be understood that similar types of film coatings or polymeric products from other companies may be used.

A lozenge containing the active ingredient can be prepared, for example, by compressing or molding the active ingredient, optionally with one or more additional ingredients. Compressed tablets may be prepared by compressing in a suitable device a free-flowing form of the active ingredient such as a powder or granules, optionally mixed with one or more binders, lubricants, excipients, surface active and dispersing agents. Molded lozenges can be made by molding in a suitable device a mixture of the active ingredient, a pharmaceutically acceptable carrier, and at least a liquid sufficient to wet the mixture. Pharmaceutically acceptable excipients for the manufacture of lozenges include, but are not limited to, inert diluents, granulating and disintegrating agents, binders and lubricants. Known dispersants include, but are not limited to, potato starch and sodium hydroxymethyl starch. Known surfactants include, but are not limited to, sodium lauryl sulfate. Known diluents include, but are not limited to, calcium carbonate, sodium carbonate, lactose, microcrystalline cellulose, calcium phosphate, dibasic calcium phosphate, and sodium phosphate. Known granulating and disintegrating agents include, but are not limited to, corn starch and alginic acid. Known binders include, but are not limited to, gelatin, acacia, pregelatinized cornstarch, polyvinylpyrrolidone, and hydroxypropylmethylcellulose. Known lubricants include, but are not limited to, magnesium stearate, stearic acid, silica, and talc.

Granulation techniques are well known in the pharmaceutical arts for modifying starting powders or other granular materials of active ingredients. Powders are usually mixed with binder material into larger permanent free-flowing agglomerates or granules called "granulation". For example, a general feature of the "wet" granulation process using solvents is that the powder is mixed with a binder material and wetted under certain conditions with water or an organic solvent to form a wet granulated material, from which the solvent must then be evaporated .

Melt granulation generally involves the use of materials that are solid or semi-solid at room temperature (ie, have a relatively low softening or melting point range) to facilitate the preparation of powders or other materials with substantially no addition of water or other liquid solvents. grain. When heated to a temperature in the melting point range, the low melting point solid liquefies to serve as a binder or granulation medium. The liquefied solid spreads itself on the surface of the powdered material with which it comes into contact, and upon cooling forms a solid particulate mass in which the initial mass is bound together. The resulting melt granulation can then be supplied to a tablet press or packaged to prepare an oral dosage form. Melt granulation improves the dissolution rate and bioavailability of active substances (ie, drugs) by forming solid dispersions or solid solutions.

US Patent No. 5,169,645 discloses directly compressible wax-containing particles with improved flow characteristics. Granules are obtained when the wax is mixed in the melt with certain flow-improving additives, and the mixture is then cooled and granulated. In certain embodiments, only the wax itself is melted in the molten composition of the wax(s) and the additive(s), and in other cases, the wax(s) and the additive(s) are melted Both will melt.

The present invention also includes multi-layered lozenges comprising layers that provide delayed release of one or more compounds useful in the methods of the present invention, and other layers that provide immediate release of one or more compounds useful in the methods of the present invention layer. Using wax/pH-sensitive polymer mixtures, it is possible to obtain gastric insoluble compositions in which the active ingredient is entrapped, thereby ensuring its delayed release.

Liquid preparations for oral administration may be in the form of solutions, syrups or suspensions. Liquid preparations can be prepared in conventional manner with pharmaceutically acceptable additives such as suspending agents (for example, sorbitol syrup, methyl cellulose, or hydrogenated edible fats); emulsifiers (for example, lecithin or acacia); non-aqueous Vehicle (eg, almond oil, oily esters, or ethanol); and preservatives (eg, methyl or propyl paraben or sorbic acid) are prepared. Liquid formulations of the pharmaceutical compositions of the present invention suitable for oral administration can be prepared, packaged and sold in liquid form or as a dry product intended for reconstitution with water or another suitable vehicle before use.

parenteral administration

As used herein, "parenteral administration" of a pharmaceutical composition includes any route of administration characterized by physical destruction of a subject's tissue and administration of the pharmaceutical composition by destruction in the tissue. Parenteral administration thus includes, but is not limited to, administration of the pharmaceutical composition by injection of the composition, administration of the composition through a surgical incision, administration of the composition through a non-surgical wound that penetrates tissue, and the like. In particular, parenteral administration is contemplated including, but not limited to, subcutaneous, intravenous, intraperitoneal, intramuscular, intrasternal injection, and renal dialysis infusion techniques.

Formulations of pharmaceutical compositions suitable for parenteral administration include the active ingredient in combination with a pharmaceutically acceptable carrier such as sterile water or sterile isotonic saline. Such formulations can be prepared, packaged or sold in a form suitable for bolus or continuous administration. Injectable formulations can be prepared, packaged, or sold in unit dosage form, eg, in ampoules or in multi-dose containers with a preservative. Injectable formulations can also be prepared, packaged, or sold in a device such as a patient-controlled analgesia (PCA) device. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, ointments, and implantable sustained-release or biodegradable formulations. Such formulations may further contain one or more additional ingredients including, but not limited to, suspending, stabilizing or dispersing agents. In one embodiment of the formulation for parenteral administration, the active ingredient is provided in dry (ie, powder or granule) form for reconstitution with a suitable vehicle (eg, sterile pyrogen-free water) prior to parenteral administration pre-reconstituted composition.

Pharmaceutical compositions can be prepared, packaged, or sold as sterile injectable aqueous or oily suspensions or solutions. This suspension or solution may be formulated according to known techniques, and may contain, in addition to the active ingredient, additional ingredients such as dispersing, wetting, or suspending agents described herein. Such sterile injectable preparations can be prepared using nontoxic parenterally acceptable diluents or solvents such as, for example, water or 1,3-butanediol. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or diglycerides. Useful other parenterally administrable formulations include those comprising the active ingredient in microcrystalline form in recombinant human albumin, fluidized gelatin, liposomal formulations, or a component of a biodegradable polymer system. Compositions for sustained release or implantation may include pharmaceutically acceptable polymers or hydrophobic materials such as emulsions, ion exchange resins, sparingly soluble polymers or sparingly soluble salts.

local administration

An obstacle to the topical administration of pharmaceutical formulations is the stratum corneum of the epidermis. The stratum corneum is a highly resistant layer composed of proteins, cholesterol, sphingolipids, free fatty acids, and various other lipids, and includes keratinocytes and living cells. One of the factors limiting the penetration (flux) of a compound across the stratum corneum is the amount of active that can be loaded or applied to the skin surface. The greater the amount of active substance applied per unit area of skin, the greater the concentration gradient between the skin surface and the underlying layers of the skin, and thus the greater the diffusivity of the active substance across the skin. Thus, formulations containing higher concentrations of active were more likely to result in more active penetration across the skin at a more consistent rate than other formulations with lower concentrations, all else being equal.

Formulations suitable for topical administration include, but are not limited to, liquid or semi-liquid formulations, such as liniments, lotions, oil-in-water or water-in-oil emulsions, such as creams, ointments, or pastes, and solutions or suspensions. Topically administrable formulations may, for example, comprise from about 1% to about 10% (w/w) active ingredient, although the concentration of the active ingredient may be the same as the solubility limit of the active ingredient in the solvent. Formulations for topical administration may further include one or more of the additional ingredients described herein.

Penetration enhancers can be used. These materials increase the penetration rate of the drug through the skin. Typical accelerators in the art include ethanol, glycerol monolaurate, PGML (polyethylene glycol monolaurate), dimethylsulfoxide, and the like. Other accelerators include oleic acid, oleyl alcohol, ethoxyethylene glycol, azone, alkanecarboxylic acids, dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone.

One acceptable vehicle for topical delivery of some compositions of the present invention may comprise liposomes. The composition of liposomes and their uses are known in the art (ie, US Patent No. 6,323,219).

In alternative embodiments, the topically active pharmaceutical composition may optionally be combined with other ingredients such as adjuvants, antioxidants, chelating agents, surfactants, foaming agents, wetting agents, emulsifiers, viscosity enhancers, buffers agents, preservatives, etc. In other embodiments, a penetration or penetration enhancer is included in the composition and is effective for improving the penetration of the active ingredient into the skin and across the stratum corneum relative to a composition lacking the penetration enhancer. Various penetration enhancers, including oleic acid, oleyl alcohol, ethoxylated glycol, laurofenone, alkanecarboxylic acids, dimethylsulfoxide, polar lipids, or N-methyl-2-pyrrolidone are in the art known to the skilled person. In another aspect, the composition may further comprise a hydrotrope, which acts to increase symptoms of the stratum corneum structure, and thus allow for increased transport across the stratum corneum. Various hydrotropes such as isopropanol, propylene glycol or sodium xylene sulfonate are known to those skilled in the art.

Topically active pharmaceutical compositions should be administered in amounts effective to effect the desired change. As used herein, an "effective amount" refers to an amount sufficient to cover the surface area of the skin in need of modification. The active compound should be present in an amount from about 0.0001% to about 15% by weight of the composition. For example, it should be present in an amount of about 0.0005% to about 5% of the composition; for example, it should be present in an amount of about 0.001% to about 1% of the composition. Such compounds may be of synthetic or natural origin.

Administered

The pharmaceutical compositions of the present invention can be prepared, packaged, or sold as formulations suitable for buccal administration. Such formulations may, for example, be in the form of lozenges or throat lozenges prepared using conventional methods, and may contain, for example, 0.1 to 20% (w/w) active ingredient, the balance comprising orally dissolvable or degradable compositions, and optionally one or more additional ingredients described herein. Alternatively, formulations suitable for buccal administration may include powdered or aerosolized or atomized solutions or suspensions containing the active ingredient. When dispersed, such powdered, aerosolized or sprayed formulations may have an average particle or droplet size in the range of about 0.1 to about 200 microns, and may further include one or more additional ingredients described herein . The examples of formulations described herein are not exhaustive, and it is to be understood that the present invention includes additional modifications of these and other formulations not described herein but known to those skilled in the art.

Rectal administration

The pharmaceutical compositions of the present invention can be prepared, packaged or sold in formulations suitable for rectal administration. Such compositions may be in the form of, for example, suppositories, retention enemas, and solutions for rectal or colonic irrigation.

Suppository formulations can be prepared by mixing the active ingredient with a non-irritating pharmaceutically acceptable excipient that is solid at normal room temperature (ie, about 20°C) and will It is liquid at rectal temperature (ie, about 37°C in healthy humans). Suitable pharmaceutically acceptable excipients include, but are not limited to, cocoa butter, polyethylene glycols and various glycerides. Suppository formulations may further contain various additional ingredients including, but not limited to, antioxidants and preservatives.

Retention enema formulations or solutions for rectal or colonic irrigation can be prepared by mixing the active ingredient with a pharmaceutically acceptable liquid carrier. As is well known in the art, the enema formulation can be administered using a delivery device appropriate to the subject's rectal anatomy, and the enema formulation can be packaged in the delivery device. Enema formulations may further contain various additional ingredients including, but not limited to, antioxidants and preservatives.

Alternative Administering Forms

Additional dosage forms of the present invention include dosage forms as described in US Pat. Nos. 6,340,475, 6,488,962, 6,451,808, 5,972,389, 5,582,837, and 5,007,790. Additional dosage forms of the present invention also include dosage forms as described in US Patent Application Nos. 20030147952, 20030104062, 20030104053, 20030044466, 20030039688, and 20020051820. Additional dosage forms of the invention are also included as described in PCT Application Nos. WO 03/35041, WO 03/35040, WO 03/35029, WO 03/35177, WO 03/35039, WO 02/96404, WO 02/32416, WO 01 Dosage forms described in WO 97783, WO 01/56544, WO 01/32217, WO 98/55107, WO 98/11879, WO 97/47285, WO 93/18755 and WO 90/11757.

Controlled Release Formulations and Drug Delivery Systems :

In certain embodiments, the compositions and/or formulations of the present invention may be, but are not limited to, short-term, fast-compensating, and controlled, eg, sustained-release, delayed-release, and pulsed-release formulations.

The term sustained release in its conventional sense refers to a pharmaceutical formulation that can gradually release a drug over an extended period of time, although not necessarily, resulting in a substantially constant blood level of the drug over an extended period of time. This time period can be as long as a month or more, and should be longer than the same amount of release administered as a bolus.

For sustained release, the compounds can be formulated with suitable polymeric or hydrophobic materials that provide the compounds with sustained release properties. Thus, the compounds used in the methods of the present invention may be administered in particulate form by, for example, injection, or in wafer or disc form by implantation.

In certain embodiments of the present invention, a compound useful in the present invention, alone or in combination with another pharmaceutical formulation, is administered to a subject using a sustained release formulation.

The term delayed release is used herein in its conventional sense to refer to a pharmaceutical formulation that provides initial release of the drug after a certain delay following drug administration, and, although not required, can include from about 10 minutes up to about 12 hours.

The term pulsatile release is used herein in its conventional sense to refer to a pharmaceutical formulation that provides drug release in a manner that produces a pulsatile plasma profile following drug administration.

The term immediate release in its conventional sense refers to a pharmaceutical formulation that provides for release of the drug immediately after drug administration.

As used herein, short term refers to any period of time following drug administration, up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, About 2 hours, about 1 hour, about 40 minutes, about 20 minutes, or about 10 minutes and any or all full or partial increments thereof.

As used herein, rapid compensation refers to any period of time following drug administration, up to and including about 8 hours, about 7 hours, about 6 hours, about 5 hours, about 4 hours, about 3 hours, about 2 hours, About 1 hour, about 40 minutes, about 20 minutes, or about 10 minutes, and any full and partial increments thereof.

Using no more than routine experimentation, those skilled in the art will recognize, or be able to ascertain, many equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents are considered to be within the scope of this invention and are covered by the appended claims. For example, it should be understood that using art-recognized alternatives and using only routine experimentation, including but not limited to reaction times, reaction sizes/volumes, and experimental reagents such as solvents, catalysts, pressures, atmospheric conditions such as nitrogen atmosphere, and reducing agents/ Modifications of the reaction conditions of the oxidant are all within the scope of this application.

The following examples further illustrate aspects of the invention. However, they are in no way limiting of the teachings or disclosures of the invention described herein.

Example

The invention will now be described with reference to the following examples. These examples are provided for illustrative purposes only, and the present invention is not limited to these examples, but includes all modifications that are obvious in light of the teachings provided herein.

Characterization method

X-ray Powder Diffraction (XRPD)

Powder X-ray diffraction was performed with a Stoe Stadi P diffractometer equipped with a Mythen 1K detector operating with Cu-K _α1 radiation. Measurements with this instrument were performed in transmission at a tube voltage of 40 kV and a tube power of 40 mA. A curved Ge monochromator allows testing with Cu-K _α1 radiation. The following parameters were set: 0.02° 2θ step size, 12 s step time, 1.5–50.5° 2θ scan range, and 1° 2θ detector step size (detector mode in step scan). For typical sample preparation, approximately 10 mg of sample was placed between two acetate foils and mounted into the Stoe transmission sample holder. During the measurement, the sample is rotated. All sample preparation and measurements were performed in an ambient air atmosphere.

Thermogravimetric Infrared Spectroscopy (TG-FTIR)

Thermogravimetric measurements were performed using a Netzsch Thermo-Microbalance TG 209 coupled to a Bruker FTIR Spectrometer Vector 22 (sample pan with pinhole, _N2 atmosphere, heating rate 10 K/min).

Microscopy

Light microscopy was performed on a Leitz Orthoplan polarizing microscope part #130880, typically at 10×10 magnification.

Nuclear Magnetic Resonance ( ¹ H-NMR)

Compounds of the present disclosure were characterized by ¹ H-NMR spectroscopy using at least one NMR spectrometer with an operating frequency selected from 300 MHz, 400 MHz, and 600 MHz; solvent peaks for reference; chemical shifts reported on the TMS scale .

Differential Scanning Calorimetry (DSC)

Differential scanning calorimetry was performed with a TA Instruments Q2000 instrument (sample pan with vacuum on lid, heating rate 10 K/min). Melting point is understood as the peak maximum.

Dynamic Vapor Sorption (DVS)

DVS measurements were performed with a SPS11-100n "Sorptions Prüsystem" from ProUmid (formerly "Projekt Messtechnik"), August-Nagel-Str. 23, 89079 Ulm (Germany). A sample of approximately 5-20 mg was placed in an aluminum sample pan. Use a humidity change rate of 5% per hour.

Place the sample on an aluminum or foil holder above the microbalance and equilibrate at 50% RH, then start the predefined humidity program:

2 hours at 50% RH

50 to 95% RH (5%/h); 5 hours at 95% RH

95 to 0% RH (5%/h); 5 hours at 0% RH

0 to 95% RH (5%/h); 5 hours at 95% RH

95 to 50% RH (5%/h); 2 hours at 50% RH

Hygroscopicity is classified based on mass gain at 85% RH relative to initial mass as follows: deliquescent (sufficient water is adsorbed to form a liquid), very hygroscopic (≥15% mass gain), absorbent Hygroscopic (<15% but ≥2% mass gain), slightly hygroscopic (<2% but ≥0.2% mass gain), or non-hygroscopic (<0.2% mass gain).

Raman spectroscopy

FT-Raman spectra were recorded on a Bruker MultiRAM FT-Raman system using a near-infrared Nd:YAG laser operating at 1064 nm and a liquid nitrogen-cooled germanium detector. 64 scans with a resolution of 2 cm ^-1 were accumulated in the range of 3500 to -50 cm ^-1 ; however, only data above 100 cm ^-1 were evaluated due to filter cutoff effects. The nominal laser power is usually 100 or 300 mW.

Solubility

Approximate solubility was determined by adding solvent in approximately 10 mg increments of compound. Solubility is expressed as <1 mg/ml if a total of at least 10 mL of solvent is added without dissolving the substance. Due to the experimental error inherent in this method, this solubility value is intended to be considered a rough estimate and is used only for the design of crystallization experiments.

Example 1 : (X) Synthesis

Step 1—Synthesis of 4,5 -difluoro -2-(2 -oxypropyl ) benzoic acid ( B)

To 2-bromo-4,5-difluorobenzoic acid (Compound A , 400 g, 1.0 eq), CuBr (24 g, 0.1 eq) and acetone (338 g, 2.0 eq) in EtOH (4 L, 10 To the suspension in vol) was added a solution of 21% NaOEt in EtOH (1.75 L, 3.0 eq) dropwise. The mixture was refluxed at 80°C for 14 hours. An LCMS sample of the reaction mixture showed one major peak with the expected mass. The mixture was cooled to room temperature (rt), filtered through CELITE®, and washed with a small amount of EtOH. The filtrate was concentrated in vacuo. The residue was diluted with dichloromethane (DCM, 15 vol) and washed with 2 N HCl (5 vol, pH=2); the aqueous layer was back extracted with DCM (2 x 600 mL). The combined organic layers _were dried over _Na2SO4 , filtered and concentrated in vacuo. The crude product was triturated with hexane (3 vol), filtered and dried to give 330 g (90.9% yield) of compound (B) . ¹ H NMR (400 MHz, chloroform- d ) δ 8.04 - 7.92 (m, 1H), 7.04 (t, J = 8.9 Hz, 1H), 4.09 (s, 2H), 2.27 (s, 3H).

Step 2— Synthesis of 6,7 -difluoro - 3 -methyl -1H- isochromen- 1 -one (C)

The above 4,5-difluoro-2-(2-oxypropyl)benzoic acid (Compound B , 330 g, 1.0 eq) was dissolved in DCM (3.3 L, 10 vol) and added slowly at room temperature Concentrated _{H2SO4 (} 33 mL). The reaction was stirred at room temperature for 14 hours. LCMS of the reaction mixture showed 1 main peak with the expected product mass. The reaction was cooled to 5°C and saturated _NaHCO3 (5 vol) was added slowly (exothermic gas evolution was observed). The mixture was stirred at room temperature for 30 minutes until the outgassing subsided. The phases settle and separate. The organic layer was washed with brine. The organic layer was dried _{over Na2SO4} , filtered and concentrated. The crude product was triturated with hexanes (3 vol), filtered and dried to give 274 g (90.7% yield) of 6,7-difluoro-3-methyl-1H-isochromen-1-one as a white solid ( C) . ¹ H NMR (400 MHz, chloroform- d ) δ 8.02 (dd, J = 10.0, 7.9 Hz, 1H), 7.12 (dd, J = 10.1, 7.1 Hz, 1H), 6.20 (s, 1H), 2.28 (s , 3H).

Step 2' —6,7 -difluoro - 3 -methyl -1H- isochromen- 1 -one (C) and methyl 4,5 -difluoro -2-(2 - oxypropyl ) benzoate ( Synthesis of C')

Compound B (221 mg) was dissolved in MeOH (3 mL) and concentrated _H2SO4 ( ₁ mL) was added slowly at room temperature. The reaction was heated at 60°C overnight. LCMS of the reaction mixture showed two main peaks, one with the expected product mass. The reaction was cooled to room temperature and slowly added to a mixture of MeOH (10 mL) and saturated NaHCO ₃ (10 mL) at 0 °C (exotherm and vigorous exhaust observed). The mixture was stirred for 30 minutes until the outgassing subsided. The mixture was concentrated in vacuo until most of the water remained. The suspension was dissolved in EtOAc and water. The phases were separated and the aqueous phase was back extracted twice with EtOAc. The organic layers were combined, dried _{over Na2SO4} , filtered and concentrated to give a yellow semisolid/oil. The crude product was loaded onto a 4 g silica gel column and eluted with 0-100% EtOAc in hexanes. The pure product filtrate (second peak from ISCO) was concentrated to give 63 mg (48% yield) of 4,5-difluoro-2-(2-oxypropyl)benzoic acid as a clear, yellow oil Methyl ester (C') . ¹ H NMR (400 MHz, chloroform- d ) δ 7.88 (dd, J = 11.0, 8.2 Hz, 1H), 6.99 (ddd, J = 10.6, 7.5, 0.4 Hz, 1H), 4.07 (s, 2H), 3.84 (s, 4H), 2.30 (s, 3H). The first peak filtrate leaving the ISCO was concentrated to give 38 mg (29% yield) of 6,7-difluoro-3-methyl- 1H -isochromen-1-one (C) as a white solid. LCMS: m/z: C10H6F2O2 ^{: M+1 calcd: 197.04 found: 197.05 along with 1} _{H NMR showed 6,7} -difluoro-3-methyl-1H-isochromene- 1-keto. ¹ H NMR (400 MHz, chloroform- d ) δ 8.03 (dd, J = 10.0, 7.9 Hz, 1H), 7.12 (dd, J = 10.1, 7.1 Hz, 1H), 6.20 (s, 1H), 2.29 (s , 3H).

Step 3 - Synthesis of 4- Acetyl- 6,7 -difluoroisoquinolin- 1(2H) -one (D)

To a solution of 6,7-difluoro-3-methyl-1H-isochromen-1-one (Compound C , 280 g, 1.0 eq) in 2-methyltetrahydrofuran (MeTHF, 2.8 L, 10 vol) K2CO3 ₍ 749 _g , 3.8 eq) was added and the system was stirred at room temperature. Formamide (193 g, 3.0 eq) was added dropwise over a 30 minute period and an exothermic reaction was observed. The reaction mixture was heated at 55-60°C for 14 hours. An LCMS sample of the reaction mixture showed one major peak with the expected product mass. The reaction mixture was cooled to 25°C and filtered. The solid was slurried with ice water (10 vol) at 0°C, then the pH was slowly adjusted to pH 6 using 2 N HCl (4.3 L) over 1.5 hours. The light brown slurry was stirred for 1 hour, filtered and washed with water, cold MeOH and hexanes, and dried to give 200 g (63% yield) of 4-acetoxy-6,7-difluoro as an off-white solid Isoquinolin-1(2H)-one (D) . ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 12.19 (s, 1H), 8.92 – 8.81 (m, 1H), 8.24 (s, 1H), 8.12 – 8.02 (m, 1H), 2.48 (s, 3H) ). Note: Tris is used in place of formamide; an alternative base such as NaOMe can be used; and a solvent such as MeOH can be used.

Step 3'— Synthesis of 4- acetyl -6,7 -difluoroisoquinolin- 1(2H) -one (D)

To methyl 4,5-difluoro-2-(2-oxypropyl)benzoate (compound C' , 63 mg, 0.28 mmol) and tris(27 mg, 0.33 mmol) in MeOH (2 mL) To the solution of 25% NaOMe in MeOH (298 mg, 1.38 mmol) was added. The mixture was stirred at room temperature for 1 hour. An LCMS sample of the reaction mixture showed one major peak with the expected product mass. After 2 hours, the reaction was quenched with 5 mL of 10% ammonium chloride solution and stirred for 15 minutes. The mixture was concentrated in vacuo to remove MeOH. The slurry was then dissolved with EtOAc and water. The phases were separated and the aqueous phase was back extracted with EtOAc. The combined organic phases were dried over Na ₂ SO ₄ , filtered and concentrated to give 58.6 mg (95% yield) of 4-acetyl-6,7-difluoroisoquinoline-1 (2H) as an off-white solid )-ketone (D) .

Step 3" — Synthesis of 4- acetyl -6,7 -difluoroisoquinolin- 1(2H) -one (D)

To a solution of 6,7-difluoro-3-methyl-1H-isochromen-1-one (Compound C , 38 mg, 0.19 mmol) and tris(19 mg, 0.23 mmol) in MeOH (1 mL) A solution of 25% NaOMe in MeOH (209 mg, 0.97 mmol) was added. The mixture was stirred at room temperature for 1 hour. An LCMS sample of the reaction mixture showed one major peak with the expected product mass. After 2 hours at room temperature, the reaction was quenched with 5 mL of 10% ammonium chloride solution and stirred for 15 minutes. The mixture was concentrated in vacuo to remove MeOH. The slurry was then dissolved with EtOAc and water. The phases were separated and the aqueous phase was back extracted with EtOAc. The combined organic phases were dried over Na ₂ SO ₄ , filtered and concentrated to give 32.3 mg (75% yield) of 4-acetyl-6,7-difluoroisoquinoline-1 (2H) as an off-white solid )-ketone (D) .

Step 4 - Synthesis of 1-(1- chloro -6,7 -difluoroisoquinolin- 4 -yl ) ethane - 1 -one (E)

To a suspension of 4-acetyl-6,7-difluoro-2H-isoquinolin-1-one (Compound D , 650 g, 2.91 mol) in acetonitrile (CAN, 11 L, 17 V) at room temperature Triethylamine (TEA, 406 mL, 2.95 mol, 1.00 eq) was added to the solution. _POCl3 (327 mL, 3.50 mol, 1.20 eq) was added slowly dropwise, at which point an exotherm of 25 to 32 °C was observed and jacket cooling should be applied. After the addition was complete, the reaction mixture was heated to an internal batch temperature of 75°C for 18 hours. Sampling of the reaction by HPLC showed 0.92% starting material, 97.22% desired product, and 0.74% later eluting peaks (presumed to be overchlorinated by-products). The reaction was cooled to 40°C and quenched very slowly with 6.6 L of water. The reaction was stirred at 40°C for 1 hour. The reaction mixture was cooled to 20°C and stirred for 1 hour, then cooled again to 0°C and aged at 0°C for 1 hour. Filter the slurry. The solid was washed with 2 L of water and then 1.5 L of ACN cooled to 0°C. The solid was vacuum dried in an oven at 40-45°C overnight to give 660 g (94% yield) of compound (E) as a tan solid. ¹ H NMR (400 MHz, chloroform- d ) δ 8.95 (dd, J = 12.4, 8.0 Hz, 1H), 8.85 (s, 1H), 8.19 (dd, J = 10.5, 8.0 Hz, 1H), 2.77(s , 3H). Note: Equivalents of POCl, addition _of base and temperature are important parameters. Larger amounts _of POCl and higher temperatures result in high levels of impurities.

Step 5 - Synthesis of 1-(6,7 -Difluoro - 1 -methoxyisoquinolin- 4 -yl ) ethane - 1 -one (F)

To 1-(1-chloro-6,7-difluoro-4-isoquinolinyl)ketene (Compound E , 965 g, 3.99 mol) in 14.5 L (15 V) MeOH and 14.5 L (15 V) THF A solution of 25% NaOMe in MeOH (1.29 L, 5.99 mol, 1.5 eq) was slowly added to the slurry in , maintaining the internal batch temperature at 0-3 °C. The solids did not completely dissolve, but produced a yellow slurry. The reaction mixture was stirred at 0°C. The reaction was sampled by HPLC after 1 hour, at which point the desired product was observed to be the main peak with 1.1% of the disubstituted impurity (dimethoxy impurity and SM co-eluting on the HPLC method originally used). ). After no more than 2 hours, the reaction was slowly quenched with 10% citric acid in water (14 L). An exotherm was observed and the white slurry was stirred at 0°C for 1 hour. The slurry was filtered and the solids were washed with water to provide a wet cake. The wet cake was vacuum dried in an oven at 35-40°C for 64-80 hours to give 881 g (93% yield) of compound (F) as an off-white fluffy solid. ¹ H NMR (400 MHz, chloroform- d ) δ 9.01 (dd, J = 13.1, 8.2 Hz, 1H), 8.74 (s, 1H), 8.02 (dd, J = 10.6, 8.3 Hz, 1H), 4.19 (s , 3H), 2.70 (s, 3H).

Step 6 - (R)-N-((R)-1-(6,7 -Difluoro - 1 -methoxyisoquinolin- 4 -yl ) ethyl )-2 -methylpropane -2- ylidene Preparation of Sulfonamide (H)

Combine ( R )-2-methylpropane-2-sulfinamide (Compound G , 495.1 g, 4085.0 mmol, 1.9 eq), 1-(6,7-difluoro-1-methoxyisoquinoline- 4-yl)Ethan-1-one (Compound F , 510.0 g, 2150.0 mmol, 1.0 eq), MeTHF (2.6 L, 5.0 V) and Ti(OEt) ₄ (1471.3 g, 6450.0 mmol, 3 eq) were added to in the reactor. The mixture was heated to 82°C and stirred. The reaction mixture was concentrated to about 3V after 1.5 hours. Add fresh MeTHF (2 V) and continue heating. This process was repeated twice over the next 3-5 hours until the reaction was deemed complete by HPLC, with a residual level of (F) not exceeding (NMT) 8% observed by HPLC. The reaction mixture was adjusted to 50 °C, MeTHF (5.1 L, 10 V) and EtOH (495.3 g, 5.0 eq) were added and then cooled to -18 °C. NaBH4 ₍ 81.3 g, 1.0 eq) was added to the reactor in 10 portions over 1 hour while maintaining the temperature between -18°C and -15°C. The mixture was stirred for about 4 hours until the reaction was deemed complete, where NMT 1% of the imine intermediate was observed by HPLC. In another reactor, lemons were prepared by adding citric acid monohydrate (1672.8 g, 3.28 S), purified water (8.0 L, 15.72 V) and sodium hydroxide (255 g, 0.5 S) to the reactor Sodium solution. The sodium citrate solution was cooled to 10°C and the reaction mixture was transferred to the sodium citrate solution with stirring while maintaining the temperature between 10°C and 16°C. The mixture was then adjusted to 25°C and stirred for about 18 hours. The layers were separated and the organic layer was washed sequentially with purified water (2.5 L), 8.0 wt% _NaHCO3 (2.5 L) solution and 24.0 wt% brine solution (2.5 L). The organic layer was dried _{over Na2SO4} and filtered. The filtrate was evaporated to about 2 V while maintaining an internal temperature of about 30°C to 36°C. The residue was subjected to two solvent exchanges with cyclopentyl methyl ether (CPME, 1.5 L, 3V). To the resulting residue was slowly added n-heptane (3.0 L, 6 V) and the mixture was heated to 50°C to 60°C for 2 hours, cooled to 20°C to 30°C over 2 hours, and stirred at this temperature 2 hours, then filtered and washed with n-heptane (3V). The solid was dried at 45°C to obtain compound (H) (584.6 g, 79.4%) as a pale yellow solid. The diastereomeric ratio (dr) of the obtained compound (H) was 97:3. m/z 343.2 [M+H] ⁺ . ¹ H NMR (400 MHz, chloroform- d ) δ 8.11 – 7.99 (m, 2H), 7.91 (dd, J = 12.0, 7.6 Hz, 1H), 5.03 – 4.93 (m, 1H), 4.11 (s, 3H) , 3.48 (s, 1H), 1.69 (d, J = 6.6 Hz, 3H), 1.23 (s, 9H). Alternatively, the crude material of Compound H can be used in the next step without further purification.

Step 7 - (R)-N-((R)-1-(6,7 -Difluoro - 1 -methoxyisoquinolin- 4 -yl ) ethyl )-N,2 - dimethylpropane- Preparation of 2 -sulfinamide (I)

Compound (H) (480 g, 1401.9 mmol) and MeTHF (6.72 L, 14 V) were added to the reactor. The mixture was stirred to obtain a clear solution, then cooled to -6°C. KOH powder (463.0 g, 7009.35 mmol, 5.0 eq) was added and stirred for 30 minutes. MeI (458.0 g, 3224.3 mmol, 2.3 eq) was added slowly over 1 hour while keeping the temperature below 0 °C. The reaction mixture was stirred at this temperature for 30 minutes and then warmed to 20°C to 25°C. The reaction mixture was stirred for 13 hours until the reaction was deemed complete, where NMT 1% of Compound (H) was observed by HPLC. The reaction mixture was cooled to 0°C to 5°C and purified water (3.84 L, 8 V) was added slowly over 2 hours. The mixture was warmed to 20°C to 25°C and stirred for 10 minutes. The aqueous layer was separated and back-extracted twice with MeTHF (2.4 L, 5 V). The organic layers were poured together and then washed with 5.0 wt% brine solution (2.4 L, 5 V) followed by purified water (2.4 L, 5 V). The organic layer was dried _over anhydrous _Na2SO4 and filtered. The filtrate was concentrated to about 3 V and solvent exchanged twice with MTBE (3.84 L, 8 V) to about 2 V. MTBE was added to the residue to adjust the volume to 3 V and the mixture was heated to 45°C. Next, n-heptane (2.88 L, 6V) was added and the mixture was heated at 45°C for 1 hour, then slowly cooled to 0°C over 2 hours. The solids were filtered and washed with 10% MTBE/n-heptane (1.44 L, 3 V). The solid was dried at 45°C to obtain Compound 1 (386.6 g, 77.5 %) as an off-white solid. The diastereomeric ratio (dr) of the obtained compound (I) was 99.5:0.5. m/z 357.2 [M+H] ⁺ . ¹ H NMR (400 MHz, chloroform-d) δ 8.08 – 7.97 (m, 2H), 7.72 (dd, J = 12.0, 7.6 Hz, 1H), 4.68 (q, J = 6.9 Hz, 1H), 4.11 (s , 3H), 2.66 (s, 3H), 1.75 (d, J = 6.9 Hz, 3H), 1.65 (s, 1H), 0.98 (s, 9H). Compound I can optionally be crystallized from isopropyl acetate (IPAc), EtOAc and/or acetonitrile.

將甲醇(2.04 L，6.0 V)加入到反應器中並冷卻到0℃。向甲醇中加入乙醯氯(599.1 g，448.9 mmol，8.0 eq)同時保持溫度低於20℃。將該混合物在20℃下攪拌1小時。在另一個反應器中，加入化合物 (I)(340.0 g，56.11 mmol，1.0 eq)和甲醇(2.72 L，8.0 V)，攪拌10分鐘，然後冷卻到5℃。將鹽酸甲醇緩慢轉移到化合物 (I)的溶液中同時保持溫度低於30℃。將反應混合物在60℃下加熱4小時直到認為反應完成，其中藉由HPLC觀察到NMT 1%的化合物 (I)。將反應混合物冷卻到35℃並濃縮到3 V，並用MeTHF (3.4 L，10 V)進行溶劑交換並減少到2.5 V。向所得的殘餘物加入MeTHF (3.4 L，10 V)和純化水(3.4 L，10 V)，然後將系統攪拌30分鐘，並分離有機層。將水層用MTBE(1.7 L，5V)洗滌並藉由緩慢添加NaHCO ₃(205.6 g，2.85 eq)調節到~pH 7.4。將水層攪拌1小時，然後加入0.1 L的THF，並在20℃下繼續攪拌14小時。過濾所得的固體，用純化水(610 mL，2 V)洗滌，並在45℃下乾燥。獲得為米色固體的化合物 (J)(177.0 g，86.5%)。m/z 239.1 [M+H] ⁺。 ¹H NMR (400 MHz, DMSO-d6) δ 11.84 (s, 1H), 9.35 (s, 1H), 8.17 – 8.07 (m, 2H), 7.64 (d, J = 4.5 Hz, 1H), 4.75 (q, J = 6.8 Hz, 1H), 2.48 (s, 3H), 1.57 (d, J = 6.7 Hz, 3H)。 Step 8 - Preparation of (R)-6,7 -difluoro - 4-(1-( methylamino ) ethyl ) isoquinolin- 1(2H) -one (J )

Methanol (2.04 L, 6.0 V) was added to the reactor and cooled to 0 °C. To methanol was added acetyl chloride (599.1 g, 448.9 mmol, 8.0 eq) while keeping the temperature below 20 °C. The mixture was stirred at 20°C for 1 hour. In another reactor, compound (I) (340.0 g, 56.11 mmol, 1.0 eq) and methanol (2.72 L, 8.0 V) were added, stirred for 10 minutes, and then cooled to 5°C. Methanol hydrochloride was slowly transferred into the solution of compound (I) while keeping the temperature below 30°C. The reaction mixture was heated at 60°C for 4 hours until the reaction was considered complete, wherein NMT 1% of Compound (I) was observed by HPLC. The reaction mixture was cooled to 35 °C and concentrated to 3 V, and solvent exchanged with MeTHF (3.4 L, 10 V) and reduced to 2.5 V. To the obtained residue were added MeTHF (3.4 L, 10 V) and purified water (3.4 L, 10 V), then the system was stirred for 30 minutes, and the organic layer was separated. The aqueous layer was washed with MTBE (1.7 L, 5V) and adjusted to ~pH 7.4 by slow addition of _NaHCO3 (205.6 g, 2.85 eq). The aqueous layer was stirred for 1 hour, then 0.1 L of THF was added and stirring was continued at 20°C for 14 hours. The resulting solid was filtered, washed with purified water (610 mL, 2 V), and dried at 45°C. Compound (J) (177.0 g, 86.5%) was obtained as a beige solid. m/z 239.1 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO-d6) δ 11.84 (s, 1H), 9.35 (s, 1H), 8.17 – 8.07 (m, 2H), 7.64 (d, J = 4.5 Hz, 1H), 4.75 (q , J = 6.8 Hz, 1H), 2.48 (s, 3H), 1.57 (d, J = 6.7 Hz, 3H).

Step 9 - (R)-3-(3- cyano - 4 -fluorophenyl )-1-(1-(6,7 -difluoro - 1 -oxo -1,2 - dihydroisoquinoline- Preparation of 4- yl ) ethyl )-1 -methylurea (X)

Compounds (J) , (K) and MeTHF were added to the reactor. After adjusting to 15-25°C, triethylamine was added. Then, the reaction mixture was heated to 50°C (45-55°C) and stirred for NLT for 6 hours. Once the reaction was complete, the temperature was adjusted to 15°C (10-20°C), and 1 N HCl solution was slowly added to the reaction mixture while maintaining an internal temperature of 10-20°C. After phase separation, the organic layer was washed sequentially with additional 1 N HCl solution, 1 N NaOH solution (×2), purified water, 8 wt % aqueous NaHCO ₃ , and purified water. Then, the organic layer was concentrated to 4.5-5.5 V while maintaining the NMT jacket temperature of 60 °C. After addition of MeTHF, the concentration to 8-10 V was repeated. The contents were filtered through a cartridge filter and concentrated to 4.5-5.5 V while maintaining an NMT jacket temperature of 60°C. After additional concentration with MeTHF, solvent exchange with EtOAc while maintaining an internal temperature of 30-55°C. After adjusting to 4.5-6.0 V (target 5 V) by adding EtOAc, the slurry was heated to 50°C (45-55°C) and stirred for NLT at the same temperature for 2 hours. Then, the contents were slowly adjusted to 15°C (10-20°C) for NLT for 3 hours, and NLT was stirred for 3 hours. The resulting slurry was filtered and the wet cake was washed with EtOAc. After deliquoring, the filter cake was vacuum dried at NMT 55°C to give compound (X) as a white to light brown solid in 75-90% yield. ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 11.61 (d, J = 5.6 Hz, 1H), 8.63 (s, 1H), 8.06 (dd, J = 10.9, 8.5 Hz, 1H), 7.99 (dd, J = 5.8, 2.7 Hz, 1H), 7.85 (ddd, J = 9.3, 4.8, 2.8 Hz, 1H), 7.66 (dd, J = 12.7, 7.5 Hz, 1H), 7.42 (t, J = 9.1 Hz, 1H) ), 7.20 (d, J = 5.7 Hz, 1H), 5.75 (q, J = 6.8 Hz, 1H), 2.60 (s, 3H), 1.41 (d, J = 6.8 Hz, 3H).

Example 2: Preparation of (3- cyano - 4 -fluorophenyl ) phenylcarbamate (K)

5-Amino-2-fluorobenzonitrile and MeTHF were added to the reactor and the temperature was adjusted to 15-25°C. Then, pyridine was added at 15-25°C. After cooling to a temperature of -10°C to -5°C, phenyl chloroformate was slowly added to the reactor while maintaining an internal temperature of NMT of 0°C. After adjusting to 0°C (-5°C to 5°C), the reaction mixture was stirred for NLT at 0°C (-5°C to 5°C) for 1 hour until the reaction was complete. Purified water was slowly added to the reaction mixture while maintaining an internal temperature of NMT of 15°C. After addition of EtOAc, the contents were adjusted to 15-25°C and stirred for NLT for 10 minutes. After phase separation, the aqueous layer was back extracted with EtOAc. The combined organic layers were washed sequentially with 1 N HCl solution (×2), then 5 wt.% NaCl solution. The organic layer was concentrated to 4.5-5.5 V while maintaining the NMT internal temperature of 35 °C. After addition of EtOAc, the contents were concentrated again to 4.5-5.5 V. After filtration through a cartridge filter, the contents were concentrated to 1-2 V. After adjusting the volume of the contents to approximately 2.5 V by adding EtOAc, the contents were adjusted to 40°C (35-45°C). Then, n-heptane was slowly added to the contents while maintaining an internal temperature of 35-45°C. After stirring the NLT at 35-45°C for 0.5 hours, the contents were slowly adjusted to 15-20°C over 1-2 hours and the NLT was stirred at the same temperature for 2 hours. The product slurry was then filtered and the wet cake was washed with 10 v/v% EtOAc in n-heptane. After deliquoring, the filter cake was vacuum dried at NMT 40°C to give compound (K) as an off-white to tan solid in 75-95% yield. ¹ H NMR (400 MHz, chloroform- d ) δ 7.79 (dd, J = 5.5, 2.9 Hz, 1H), 7.65 (dt, J = 9.2, 3.6 Hz, 1H), 7.46 – 7.36 (m, 2H), 7.32 – 7.18 (m, 3H), 7.18 – 7.14 (m, 1H), 7.12 (s, 1H).

Example 3: (X) crystal form 1 purification and crystallization

To ( R )-6,7-difluoro-4-(1-(methylamino)ethyl)isoquinolin-1( 2H )-one hydrochloride (0.91 g, 3.31 mmol, 1.0 eq.) To a suspension in 27 mL of THF was added triethylamine (0.97 mL, 6.94 mmol, 2.1 eq) followed by phenyl N-(3-cyano-4-fluoro-phenyl)carbamate (0.85 g , 3.31 mmol, 1.0 eq). The reaction was stirred at room temperature for 5 minutes, then heated to 50°C for 5 hours. When the reaction was complete, the mixture was concentrated in vacuo and the oil was diluted with EtOAc. The organic layer was washed with 0.2 N HCl solution and the aqueous phase was back extracted with EtOAc. The combined organic layers were washed with _NaHCO3 solution, dried _{over Na2SO4} , filtered and concentrated to an oil. The oil was diluted with 3 mL of DCM at which point the product started to precipitate. The isolated solid was combined with 2 smaller batches prepared by the same method and dissolved in 10% MeOH in DCM. After drying in a vacuum oven at 50°C for 24 hours, the solvent was removed in vacuo to give 1.99 g (75.6% combined yield) of compound (X). LCMS: m/z found 401.10 [M+H] ⁺ . ¹ H NMR (400 MHz, DMSO- d ₆ ): δ 11.61 (d, J = 5.6 Hz, 1H), 8.64 (s, 1H), 8.06 (dd, J = 10.9, 8.5 Hz, 1H), 7.99 (dd , J = 5.8, 2.7 Hz, 1H), 7.85 (ddd, J = 9.3, 4.8, 2.8 Hz, 1H), 7.66 (dd, J = 12.7, 7.5 Hz, 1H), 7.42 (t, J = 9.1 Hz, 1H), 7.20 (d, J = 5.7 Hz, 1H), 5.75 (q, J = 6.8 Hz, 1H), 2.60 (s, 3H), 1.41 (d, J = 6.8 Hz, 3H).

Example 4: (X) crystal form 1 representation of

Initial characterization of the dichloromethane solvate Form 1 was performed including polarized light microscopy (PLM), X-ray powder diffraction (XRPD), thermal analysis characterization (TG-FTIR), dynamic vapor adsorption (DVS) and solubility studies.

Polarized light microscopy analysis showed that the particles were crystalline, very small and partially agglomerated (Figure 1). Hot stage microscopy was performed by heating from 25°C to 250°C at 10 K/min. Optical images were collected at several temperatures (Figures 2A-2K). Observations in this experiment indicate that the samples undergo desolvation and lose crystallinity in the temperature range of about 140°C to 158°C. However, between 170°C and 193°C, the samples appeared to recrystallize before dissolving between 217°C and 222°C. Therefore, a phase transition to the desolvated form may have occurred.

XRPD analysis of Form 1 confirmed the crystalline nature of the starting material (Figure 3). The low peak intensities and rather broad peaks may indicate poor crystallinity, however these may be attributed to the small particle size of the sample rather than inherent to the solvate itself.

surface 1. crystal form 1 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 6.5 13.7 44 2 7.1 12.5 77 3 8.1 10.9 71 4 9.6 9.2 45 5 10.8 8.2 36 6 12.1 7.3 58 7 13.0 6.8 39 8 13.2 6.7 52 9 14.3 6.2 54 10 15.5 5.71 51 11 16.5 5.37 31 12 16.9 5.24 34 13 17.3 5.12 34 14 17.7 5.02 38 15 18.3 4.84 33 16 19.1 4.65 48 17 19.5 4.55 34 18 21.0 4.22 29 19 21.4 4.14 30 20 21.9 4.05 33 twenty one 22.9 3.88 32 twenty two 24.0 3.70 32 twenty three 24.3 3.66 34 twenty four 25.3 3.52 twenty four 25 25.8 3.45 70 26 26.1 3.41 93 27 26.5 3.36 100 28 27.0 3.29 44 29 27.9 3.19 36 30 28.6 3.12 26 32 30.5 2.93 twenty four 32 32.6 2.74 34

Form 1 was further subjected to hot stage XRPD, where the sample was heated from 25°C to 185°C at 10 K/min, and XRPD patterns were collected at several temperatures (Figure 4). While the temperature varies between measurements, it remains constant during each XRPD measurement. XRPD patterns were collected at 160°C, 165°C, 170°C, 180°C and 185°C. At higher temperatures, the sample color changed from white to brown. In addition, the superimposed XRPD pattern showed that Form 1 transformed into Form 2 when the temperature reached 160 °C. At higher temperatures (>170°C) the samples start to lose crystallinity and melt.

Thermogravimetric analysis combined with FT-IR spectroscopy was performed on Form 1. TG-FTIR showed that the sample contained about 10% DCM, which corresponded well to the stoichiometric amount of hemisolvate (Figure 5). Analysis further showed that the sample decomposed at temperatures above about 200°C. The high solvent content of Form 1 was known by TG-FTIR, DSC was performed in a sample pan with pinholes, and a drying step was introduced to obtain a solvent-free sample. DSC showed a glass transition close to 130 °C with a ΔCp of about 0.4 J/(g∙K), which is typical for amorphous pharmaceutical compounds (Figure 6). At higher temperatures, a small portion of the crystallized fraction appears to melt, and an exothermic signal may correspond to recrystallization.

Dynamic vapor sorption (DVS) studies showed that Form 1 lost almost all DCM during the first cycle (Figures 7 and 8). It is unlikely that all DCMs were replaced by water by the end of the test. The XRPD patterns before and after the DVS experiment are quite similar and indicate a considerable degree of isomorphism. Conversion from solvate to hydrate occurred and this new crystal form was designated as Form 9. The most notable difference is a strong reflection at 4.0° 2θ (Figure 9). In addition, TG-FTIR of the sample after DVS (Form 9) confirmed that dichloromethane was replaced by water and the water content was about 3.8% (Figure 10). This result may indicate that the starting material may be a channel-type solvate. During the measurement, DCM was completely removed.

The approximate solubility of Form 1 was determined at room temperature. These values were obtained by adding a small aliquot of solvent to approximately 10 mg of solid to achieve dissolution by short periods of shaking and/or sonication. The maximum amount of solvent used for these assays is 10 mL. Note that these values are only approximations and do not necessarily correspond to thermodynamic solubility values.

surface 2. (X) crystal form 1 solubility studies of solvent Approximate solubility (mg/ml) 2-Propanol < 1 Acetic acid ~ 12 acetone ~ 2 Acetone: Water (4:1) ~6 DCM 6 - 8 DMA ~ 202 DMA:ethanol (1:1) 16 - 19 DMA: Ethyl acetate (1:1) 134 - 268 DMF ~ 224 DMF: Water (1:4) < 1 DMSO 63 - 94 DMSO:ethyl acetate (1:2) ~ 216 Ethanol < 1 Ethanol:Water (4:1) < 1 Ethyl acetate < 1 n-heptane < 1 methanol < 1 Methanol:DCM (1:1) ~ 7 Methyltetrahydrofuran < 1 TBME < 1 THF ~ 4

Example 5 : (X) crystal form 2 purification and crystallization

Compound (J) (1.0 kg), compound (K) (1.0 kg, 0.93 eq.) and 2-MeTHF (25.2 kg) were added to the reactor. Triethylamine (0.47 kg, 1.1 eq.) was further added to the reactor at 15-25°C, and the lines used were flushed with an additional 0.43 kg of 2-MeTHF. The reactor contents were heated to 50°C (45-55°C) for no less than 6 hours. The reaction was monitored by HPLC and was considered complete when no more than 2% (J) remained, and no more than 0.5% (K) remained.

The contents of the reactor were cooled to an internal temperature of 15°C (10-20°C), and the internal temperature was maintained as 1 N HCl solution (5.1 kg) was slowly added to the reactor. The reactor contents were stirred at 10-20°C, then the stirring was stopped, the phases were separated, and the bottom aqueous layer was discarded. 1 N HCl solution (5.1 kg) was added to the reactor again while maintaining the temperature at 10-20°C. The reactor contents were stirred at 10-20°C, then the stirring was stopped, the phases were separated, and the bottom aqueous layer was discarded. Next, 1 N NaOH solution (5.2 kg) was added to the reactor while maintaining the temperature at 10-25°C. The reactor contents were stirred at 20-25°C, then the stirring was stopped, the phases were separated, and the bottom aqueous layer was discarded. 1 N NaOH solution (5.2 kg) was added to the reactor again while maintaining the temperature at 10-25°C. The reactor contents were stirred at 20-25°C, then the stirring was stopped, the phases were separated, and the bottom aqueous layer was discarded. Next, purified water (5.0 kg) was added to the reactor. The reactor was stirred, then stopped, the phases were separated, and the bottom aqueous phase was discarded. Next, _NaHCO3 solution (5.3 kg, 8 wt.% aq.) was added to the reactor while maintaining the temperature at 20-25 °C. The reactor contents were stirred at 20-25°C, then the stirring was stopped, the phases were separated, and the bottom aqueous layer was discarded. Next, purified water (5 L) was added to the reactor, stirred, then stopped, the phases were separated, and the final aqueous layer was discarded.

The organic phase was concentrated under vacuum to 4.5-5.5 L while maintaining a jacket temperature not exceeding 60°C. Next, 2-MeTHF (8.84 kg) was added to the organic phase. The material was re-concentrated to 8-10 L under vacuum while maintaining a jacket temperature not exceeding 60°C. The organic material was Polish filtered through a cartridge filter and the filtrate was transferred to the reactor. Flush all lines and filters with 2-MeTHF (0.85 kg).

The organic phase was concentrated under vacuum to 4.5-5.5 L while maintaining a jacket temperature not exceeding 60°C, then 2-MeTHF (4.27 kg) was added. The organic phase was concentrated under vacuum to 4.5-5.5 L while maintaining a jacket temperature not exceeding 60°C, then 2-MeTHF (4.27 kg) was added. The temperature of the contents of the reactor was adjusted to 35-45°C, then the contents were concentrated under vacuum to 3.5-4.5 L while maintaining an internal temperature of 30-55°C.

Next, EtOAc (5 L) was added and the temperature of the reactor contents was adjusted to 35-45°C. The contents of the reactor were concentrated under vacuum to 3.5-4.5 L while maintaining an internal temperature of 30-55 °C, then EtOAc (5 L) was added. The temperature of the contents of the reactor was adjusted to 35-45°C. The contents of the reactor were concentrated under vacuum to 3.5-4.5 L while maintaining an internal temperature of 30-55°C. The volume of the reactor contents was adjusted to 4.5-6.0 L by adding EtOAc, then the reactor contents were stirred for no less than 5 minutes. Next, the internal temperature of the reactor was adjusted to 45-55°C and stirred for not less than 2 hours. The contents of the reactor were slowly cooled to 10-20°C for not less than 3 hours and stirred for not less than 3 hours. The resulting slurry was filtered and the wet cake was deliquored. The wet cake was washed with EtOAc (2 x 2 L) and then dried under vacuum at no more than 55°C for no less than 12 hours until residual 2-MeTHF and EtOAc were each present in amounts no greater than 5000 ppm. After this step, compound (X)(R)-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxo-1,2- Dihydroisoquinolin-4-yl)ethyl)-1-methylurea is usually obtained from compound (J) in 79-86% yield with a purity of not less than 99.0% as a white to off-white solid.

Example 6 : (X) crystal form 2 representation of

Polarized light microscopic analysis indicated that the particles were crystalline (Figure 11). Compared to Form 1, the particles of Form 2 are significantly larger and exhibit stronger birefringence. XRPD analysis further demonstrated the crystalline nature of Form 2 (Figure 12). The high crystallinity of Form 2 contrasts with the lower crystallinity observed in Form 1 (Figure 13).

surface 3. crystal form 2 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 8.26 10.70 46 2 11.29 7.83 67 3 12.02 7.35 68 4 14.27 6.20 25 5 17.22 5.15 twenty two 6 17.51 5.06 79 7 18.00 4.92 twenty three 8 18.32 4.84 31 9 19.58 4.53 30 10 20.06 4.42 twenty three 11 21.25 4.18 38 12 21.84 4.07 52 13 22.09 4.02 69 14 22.72 3.91 100 15 24.72 3.60 twenty three 16 25.19 3.53 33 17 27.49 3.24 18 18 28.15 3.17 42 19 29.54 3.02 37 20 31.74 2.82 16 twenty one 32.10 2.79 twenty one twenty two 33.06 2.71 14

TG-FTIR was performed on Form 2 and mass loss was detected up to about 200°C, indicating that this is the anhydrous polymorph of (X) (Figure 14).

Differential scanning calorimetry of Form 2 showed a rather sharp endotherm with a maximum peak at 219°C, which was attributed to melting and correlated with a melting enthalpy of about 84 J/g (Figure 15).

The behavior of Form 2 in the presence of variable water vapour pressure was investigated using dynamic vapour adsorption measurements. The results of the DVS measurements showed that the change in water content was relatively small over the entire relative humidity range (Figures 16 and 17). At 95% relative humidity, less than 0.2% water is absorbed, therefore, Form 2 is considered non-hygroscopic.

Example 7: (X) Polymorph screening of

Unless otherwise indicated, Form 1 of (X) was used as starting material for solvent-mediated polymorph screening including slurries under a number of pressure conditions in which temperature, solvent, and water activity differed experiment. New crystalline forms were observed and labeled as Forms III-VIII. The general conditions used to obtain the new crystal forms are provided in Table 4. At least one detailed description of the preparation of each crystalline form is provided herein.

Table 4. Polymorph screening of free crystalline form (1) using slurry experiments experiment solvent experiment result 1 acetone Slurry (rt) Form 3 2 Ethanol Slurry (rt) Form 4 3 Ethanol dry ^a Form 8 4 DCM Slurry (rt) Form 3 5 DMF/water precipitation NA 6 DMF/TBME precipitation Form 3 7 DCM evaporation Amorphous 8 water Slurry (rt) Form 2 9 - heating Amorphous 10 THF cool down Form 3 11 Acetone/Water (4:1, v/v) cool down Form 7 12 Acetone/Water (4:1, v/v) dry ^b mixture 13 water Slurry (rt) Form 2 14 Ethyl acetate Slurry (rt) Form 5 15 2-Methyl-THF Slurry (rt) Form 2 16 2-Propanol Slurry (rt) Form 2 17 n-heptane Slurry (temperature cycle) Form 2 18 methanol Slurry (40℃) Form 6 19 Toluene Slurry (40℃) Form 2 20 water Slurry (40℃) Form 9 twenty one DMSO/MEK/toluene cool down Form 3 ^a Form 4 was used as starting material; ^b Form 7 was used as starting material.

Example 8 : (X) crystal form 3 Crystallization and Characterization of

A sample of (X) Form 1 (54.2 mg) was dissolved in 4.6 mL of THF at 50°C. The resulting colorless solution was cooled to 20°C for 10 hours and stirred at 20°C for about 9 hours. The clear solution was then cycled between 20°C and 5°C (1 cycle), followed by cooling to 5°C (eg, 20°C to 5°C for 10 hours; 5°C to 20°C for 10 hours; 20°C to 5°C for 10 hours) Hour). After stirring at 5 °C for one day, the resulting diluted white suspension was concentrated under N2 for ₂ h, then stirring was continued at 5 °C for 2 days. Then, the suspension was filtered and centrifuged, and a white solid product was obtained.

The collected solids were analyzed by XRPD and TG-FTIR. The observed low intensity XRPD reflections for Form 3 may indicate poor crystallinity (Figure 18). TG-FTIR showed no significant mass loss (Figure 19).

surface 5. crystal form 3 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 8.34 10.59 46 2 10.78 8.20 100 3 11.28 7.84 69 4 12.49 7.08 53 5 14.37 6.16 45 6 17.65 5.02 58 7 18.65 4.75 43 8 19.10 4.64 68 9 22.66 3.92 42 10 23.20 3.83 33 11 24.03 3.70 39 12 24.76 3.59 26 13 25.27 3.52 32 14 26.45 3.37 44 15 28.37 3.14 27 16 30.52 2.93 20 17 32.67 2.74 twenty four

Example 9: (X) crystal form 4 representation of

A sample of (X) Form 1 (55.6 mg) was suspended in 0.6 mL of ethanol and the resulting white suspension was stirred at room temperature for 6 days. The suspension was filtered and centrifuged to obtain a white solid.

The collected solids were analyzed by XRPD and TG-FTIR. The XRPD pattern indicated that Form 4 was crystalline in nature (Figure 20). TG-FTIR showed that Form 4 contained approximately 18% ethanol, but no significant steps were observed, thus complicating the distinction between surface absorption and lattice bound ethanol. However, the fact that temperatures above 170°C were required to remove all solvent indicated that Form 4 was an ethanol solvate of (X) (Figure 21).

surface 6. crystal form 4 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 5.1 17.4 30 2 8.0 11.1 65 3 8.6 10.3 40 4 9.7 9.1 37 5 10.3 8.6 32 6 10.8 8.2 61 7 11.3 7.8 twenty one 8 12.5 7.1 35 9 12.9 6.8 twenty four 10 13.2 6.7 100 11 14.3 6.2 twenty one 12 15.2 5.82 20 13 16.0 5.54 18 14 16.6 5.33 18 15 16.9 5.25 39 16 17.4 5.09 twenty two 17 17.6 5.02 twenty four 18 18.4 4.81 33 19 18.7 4.74 twenty three 20 19.2 4.61 26 twenty one 20.2 4.40 16 twenty two 20.5 4.33 twenty one twenty three 21.1 4.21 twenty four twenty four 21.3 4.16 twenty four 25 21.6 4.11 twenty two 26 21.8 4.07 19 27 22.5 3.95 twenty four 28 23.0 3.86 twenty three 29 23.6 3.76 49 30 24.0 3.70 17 32 24.3 3.66 17 32 25.2 3.53 63 33 25.5 3.50 71 34 25.9 3.43 36 35 26.5 3.35 91 36 26.9 3.31 18 37 27.3 3.26 12 38 27.8 3.21 12 39 28.4 3.14 15 40 28.8 3.09 15 41 29.5 3.03 13 42 30.1 2.97 twenty four 43 30.5 2.93 12 44 31.3 2.86 10 45 31.6 2.83 11 46 32.1 2.79 11 47 32.4 2.76 12 48 32.9 2.72 12 49 33.5 2.67 9 50 33.9 2.64 9 51 34.5 2.60 9 52 35.4 2.53 10 53 35.7 2.51 8 54 36.1 2.49 7 55 36.7 2.45 8 56 38.0 2.37 8 57 38.2 2.35 9 58 39.4 2.29 8 59 39.9 2.26 7 60 40.7 2.22 6

Example 10: (X) crystal form 5 representation of

A sample of (X) Form 1 (48.5 mg) was suspended in 0.6 mL EtOAc and the resulting white suspension was stirred at room temperature for 6 days. The suspension was filtered and centrifuged to obtain a white solid.

The collected solids were analyzed by XRPD and TG-FTIR. The XRPD pattern indicated that Form 5 was crystalline in nature (Figure 22). TG-FTIR indicated that the sample contained approximately 9% EtOAc, which is close to the theoretical value of 9.9% for the hemisolvate (Figure 23).

surface 7. crystal form 5 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 4.03 21.91 44 2 6.45 13.70 42 3 7.09 12.46 100 4 8.06 10.96 95 5 9.58 9.23 59 6 12.12 7.30 60 7 13.17 6.72 52 8 14.15 6.25 60 9 15.57 5.69 70 10 15.86 5.58 66 11 17.34 5.11 39 12 19.27 4.60 37 13 19.80 4.48 34 14 20.79 4.27 29 15 21.62 4.11 35 16 22.13 4.01 33 17 22.51 3.95 30 18 23.53 3.78 28 19 24.40 3.64 34 20 25.72 3.46 71 twenty one 26.07 3.42 71 twenty two 26.42 3.37 88 twenty three 26.77 3.33 53 twenty four 27.67 3.22 47 25 27.94 3.19 27 26 28.55 3.12 twenty three 27 29.24 3.05 16 28 29.54 3.02 18 29 30.38 2.94 twenty two 30 30.73 2.91 twenty two 32 32.71 2.74 twenty three

Example 11: (X) crystal form 6 representation of

A sample of (X) Form 1 (49.6 mg) was suspended in 0.3 mL of methanol and the resulting white suspension was stirred at 40°C for 6 days. Then, the suspension was filtered and centrifuged, and a white solid was obtained.

The collected solids were analyzed by XRPD and TG-FTIR. The XRPD pattern indicated that Form 6 was crystalline in nature (Figure 24). TG-FTIR indicated that the sample contained about 6.4% methanol, which was released in the temperature range from room temperature to about 150°C (Figure 25). No distinct steps were observed, complicating the distinction between surface absorption and lattice bound methanol. However, the fact that temperatures > 100°C are required to remove all solvent indicates that Form 6 is a methanol solvate.

surface 8. crystal form 6 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 5.2 17.1 79 2 7.4 11.9 39 3 7.8 11.4 100 4 8.4 10.5 13 5 9.5 9.3 36 6 10.1 8.8 57 7 10.9 8.1 30 8 11.3 7.8 49 9 11.5 7.7 31 10 12.7 7.0 twenty two 11 13.5 6.5 16 12 13.8 6.4 96 13 14.3 6.2 16 14 14.9 5.94 11 15 15.1 5.86 16 16 15.5 5.70 19 17 16.3 5.42 9 18 16.7 5.31 13 19 17.1 5.18 12 20 17.7 5.02 18 twenty one 18.3 4.85 18 twenty two 18.7 4.75 19 twenty three 19.1 4.64 35 twenty four 19.7 4.51 12 25 20.1 4.42 10 26 20.7 4.29 11 27 21.7 4.09 19 28 21.9 4.05 19 29 22.5 3.95 26 30 22.7 3.91 28 32 23.3 3.82 15 33 24.0 3.71 11 34 24.5 3.63 14 35 25.0 3.56 13 36 25.5 3.49 twenty one 37 25.7 3.46 51 38 26.0 3.42 35 39 26.5 3.36 32 40 26.7 3.33 32 41 27.3 3.26 11 42 27.9 3.19 32 43 28.5 3.13 12 44 29.7 3.01 10 45 29.9 2.99 11 46 30.6 2.91 10 47 31.0 2.88 7 48 31.6 2.83 17 49 32.0 2.79 9 50 32.3 2.77 11 51 32.5 2.75 13 52 33.1 2.71 11 53 33.7 2.66 7 54 34.3 2.62 7 55 34.6 2.59 7 56 34.9 2.57 6 57 36.0 2.49 8 58 36.6 2.45 6 59 37.1 2.42 5 60 38.7 2.33 8 61 40.1 2.25 5 62 40.7 2.21 5 63 41.2 2.19 8

Example 12: (X) crystal form 7 representation of

A sample of (X) Form 1 (47.9 mg) was dissolved in 2.7 mL of acetone/water (4/1) at 50°C. The resulting colorless solution was cooled to 20°C for 10 hours and stirred at 20°C for about 9 hours. The clear solution was then cycled between 20°C and 5°C (1 cycle), followed by cooling to 5°C (eg, 20°C to 5°C for 10 hours; 5°C to 20°C for 10 hours; 20°C to 5°C for 10 hours) Hour). After stirring at 5 °C for one day, the resulting diluted white suspension was concentrated under N2 for ₂ h, then stirring was continued at 5 °C for 2 days. The suspension was then filtered and centrifuged, and the product was obtained as a white solid.

The collected solids were analyzed by XRPD and TG-FTIR. The XRPD pattern indicated that Form 7 was crystalline in nature (Figure 26). TG-FTIR indicated that the sample contained about 38.7% water and acetone, which were released in the temperature range from room temperature to about 130°C (Figure 27). The fact that a temperature >130°C is required to remove all solvent and the presence of a second step at about 110°C indicates that Form 7 is an acetone solvate or a mixed solvate/hydrate of (X) .

surface 9. crystal form 7 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 4.0 22.0 29 2 7.0 12.5 27 3 7.6 11.6 34 4 7.8 11.3 46 5 9.0 9.8 63 6 9.6 9.2 55 7 10.1 8.7 32 8 10.7 8.2 25 9 11.3 7.8 16 10 11.6 7.6 14 11 12.1 7.3 27 12 12.5 7.1 18 13 12.9 6.8 97 14 13.5 6.5 80 15 13.8 6.4 16 16 14.2 6.2 19 17 14.7 6.0 31 18 15.1 5.87 12 19 15.6 5.68 16 20 16.1 5.49 twenty three twenty one 16.9 5.25 twenty two twenty two 17.7 5.01 19 twenty three 18.0 4.93 12 twenty four 18.4 4.83 20 25 18.7 4.74 13 26 19.1 4.64 twenty two 27 19.5 4.56 18 28 19.9 4.46 32 29 20.4 4.34 66 30 20.7 4.29 53 32 21.0 4.22 51 33 21.5 4.13 15 34 22.3 3.99 13 35 22.6 3.93 15 36 22.9 3.87 27 37 23.3 3.81 twenty three 38 24.1 3.69 16 39 24.4 3.65 14 40 25.2 3.53 40 41 26.0 3.43 100 42 26.4 3.37 31 43 27.0 3.30 twenty four 44 27.2 3.28 27 45 27.8 3.21 36 46 28.2 3.16 twenty three 47 28.4 3.14 20 48 28.8 3.10 29 49 29.2 3.05 18 50 30.0 2.98 10 51 30.5 2.93 10 52 30.7 2.91 9 53 31.4 2.85 10 54 31.8 2.81 11 55 32.6 2.74 17 56 32.9 2.72 19 57 33.3 2.69 10 58 33.7 2.66 8 59 34.9 2.57 9 60 35.3 2.54 10 61 35.8 2.51 7 62 36.6 2.45 8 63 37.3 2.41 7 64 37.5 2.40 6 65 38.2 2.35 10 66 38.9 2.31 7 67 39.4 2.28 8 68 40.1 2.25 8 69 40.3 2.23 8 70 41.2 2.19 7

Example 13: (X) crystal form 8 representation of

A sample of Form (X) was vacuum dried at room temperature for 2 days, then the sample was vacuum dried at 40°C for one day.

Solids were analyzed by XRPD and TG-FTIR. The XRPD pattern indicated that Form 8 was crystalline in nature (Figure 28). Additionally, an overlay comparing Form 8 and Form 4 (before drying) is provided (FIG. 29). TG-FTIR analysis showed a water content of about 2.7% and the absence of ethanol (Figure 30). The observed mass loss is close to the expected theoretical water content of the hemihydrate, which is 2.25%. However, further research is needed to clarify this crystal form.

surface 10. crystal form 8 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 5.2 17.1 94 2 7.2 12.3 32 3 7.4 11.9 46 4 7.8 11.4 89 5 8.3 10.6 28 6 9.1 9.7 29 7 9.5 9.3 46 8 10.1 8.8 55 9 10.9 8.1 34 10 11.4 7.8 55 11 11.6 7.6 36 12 12.7 7.0 29 13 13.9 6.4 100 14 15.2 5.81 twenty four 15 15.6 5.68 27 16 16.7 5.30 twenty three 17 17.7 5.01 26 18 18.3 4.84 twenty three 19 18.7 4.75 25 20 19.1 4.64 33 twenty one 20.8 4.26 18 twenty two 21.8 4.08 28 twenty three 21.9 4.05 26 twenty four 22.6 3.93 30 25 22.8 3.90 31 26 23.2 3.83 19 27 24.0 3.70 18 28 25.0 3.56 19 29 25.7 3.46 49 30 26.1 3.41 33 32 26.5 3.36 27 33 26.9 3.32 36 34 27.4 3.25 15 35 28.1 3.17 32 36 30.0 2.98 12 37 30.8 2.90 13 38 31.8 2.82 20 39 32.5 2.76 15 40 33.0 2.71 12 41 33.7 2.66 11 42 34.9 2.57 9 43 35.8 2.51 10 44 36.1 2.49 9 45 38.7 2.32 9 46 41.2 2.19 9

Example 14 : (X) crystal form 9 representation of

Dynamic vapor adsorption (DVS) experiments were performed on samples of (X) Form 1, which indicated that Form 1 of (X) lost almost all DCM during the first cycle, and that DCM might be replaced by water at the end of the cycle, yielding (X) ) of the crystalline form 9.

The solid recovered after DVS was analyzed by XRPD and TG-FTIR. Although the XRPD patterns before and after DVS are similar, a considerable degree of isomorphism is still suggested (Figure 9). Form 9 may be the result of solvate to hydrate conversion. The most noticeable difference is a strong reflection at 4.0º 2θ (Figure 9). TG-FTIR of the samples after DVS confirmed the replacement of DCM by water. The water content was approximately 3.8%, indicating that Form 9 may be a channel-type solvate (Figure 10).

Form 9 of (X) was also prepared by recrystallization from water. A sample of (X) Form 1 (51.8 mg) was suspended in 0.8 mL of water. The resulting white suspension was stirred at 40°C for 6 days, and then the suspension was filtered and centrifuged to obtain a white solid.

surface 11. crystal form 9 of XRPD Peak meter Numbering Location [°2θ] d- spacing [A] strength [%] 1 4.0 22.0 100 2 6.5 13.5 41 3 7.2 12.3 79 4 8.1 10.9 76 5 9.6 9.2 52 6 10.8 8.2 35 7 11.1 7.9 33 8 12.1 7.3 45 9 13.2 6.7 38 10 14.4 6.1 57 11 15.4 5.76 37 12 15.7 5.63 36 13 16.7 5.31 26 14 17.6 5.02 28 15 19.1 4.64 34 16 22.7 3.92 25 17 23.2 3.83 twenty two 18 24.1 3.69 25 19 24.4 3.65 28 20 26.0 3.42 44 twenty one 26.4 3.37 67 twenty two 27.3 3.27 29 twenty three 28.2 3.16 28 twenty four 28.8 3.09 20 25 30.8 2.90 18 26 32.7 2.73 17

Example 15: (X) of competitive pulp

Competitive suspension equilibrium experiments were used to determine thermodynamically stable polymorphs under the conditions tested (Table 12). Two experiments were performed independently at room temperature and at 50°C with acetone as solvent. In each case, a sample of (X) Form 2 was suspended in acetone and seeded with Form 3, and the resulting suspension was stirred for 7 days at the indicated temperature (ie, rt or 50°C).

Comparison of the XRPD patterns of the two starting materials and the two products from the experiments indicated that Form 2 was a thermodynamically stable crystalline form at room temperature and 50°C (Figure 31).

surface 12. Competitive Slurry Experiment experiment solvent mixture experiment result 1 acetone Form 2 + Form 3 Slurry (r.t.) Form 2 2 acetone Form 2 + Form 3 Slurry (50℃) Form 2

List the implementation :

The following enumerated embodiments are provided, and their numbering should not be construed as specifying a level of importance.

(X)， 該方法包括使(3-氰基-4-氟苯基)胺基甲酸苯基酯 (K)：

(K)與( R)-6,7-二氟-4-(1-(甲胺基)乙基)異喹啉-1(2H)-酮 (J)：

(J)反應以便生成包含 (X)的第一反應系統。 Embodiment 1 provides the preparation of ( R )-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxo-1,2-dihydroisoquinoline) A method for olin-4-yl)ethyl)-1-methylurea (X) , or a salt or solvate thereof:

(X) , the method comprising making (3-cyano-4-fluorophenyl) phenylcarbamate (K) :

(K) with ( R )-6,7-difluoro-4-(1-(methylamino)ethyl)isoquinolin-1(2H)-one (J) :

(J) reacts to produce a first reaction system comprising (X) .

Embodiment 2 provides the method of embodiment 1, wherein the reaction of (K) with (J) is carried out in the presence of a base.

Embodiment 3 provides the method of embodiment 2, wherein the base comprises triethylamine, diisopropylethylamine, and/or pyridine.

Embodiment 4 provides the method of any one of embodiments 1-3, wherein the reaction of (K) with (J) is carried out in a solvent comprising 2-methyltetrahydrofuran.

Embodiment 5 provides the method of any one of embodiments 1-4, wherein the purification of (X) comprises neutralizing at least a portion of the base in the first reaction system and forming the resulting (X) in 2-methyltetrahydrofuran. first solution.

Embodiment 6 provides the method of embodiment 5, wherein the purification of (X) further comprises exchanging at least a portion of the 2-methyltetrahydrofuran in the first solution with ethyl acetate, thereby forming a second reaction system.

Embodiment 7 provides the method of embodiment 6, wherein the second reaction system is at least partially concentrated to produce solid (X) .

Embodiment 8 provides the method of any of embodiments 1-7, wherein (K) is prepared by reacting 5-amino-2-fluorobenzonitrile with phenyl chloroformate.

Embodiment 9 provides the method of embodiment 8, wherein 5-amino-2-fluorobenzonitrile is reacted with phenyl chloroformate in the presence of a base.

Embodiment 10 provides the method of embodiment 9, wherein the base comprises triethylamine, diisopropylethylamine, and/or pyridine.

(I)。 Embodiment 11 provides the method of any one of embodiments 1-10, wherein by making ( R )-N-(( R )-1-(6,7-difluoro-1-methoxyisoquinoline- 4-yl)ethyl)-N,2-dimethylpropane-2-sulfinamide (I) reacts with acid to prepare (J) :

(I) .

Embodiment 12 provides the method of embodiment 11, wherein the acid comprises an ethereal solution of HCl and/or a solution of acetyl chloride in methanol.

(H)。 Embodiment 13 provides the method of any one of embodiments 11-12, wherein (I) is obtained by subjecting ( R )-N-(( R )-1-(6,7-difluoro-1-methoxyiso Quinolin-4-yl)ethyl)-2-methylpropane-2-sulfinamide (H) methylation to prepare:

(H) .

Embodiment 14 provides the method of embodiment 13, wherein the methylating agent comprises methyl iodide, methyl chloride, methyl triflate, and/or dimethyl sulfate.

(G)與1-(6,7-二氟-1-甲氧基異喹啉-4-基)乙烷-1-酮 (F)

(F)反應以便形成相應的N-亞烷基亞磺醯胺，並用還原劑處理該N-亞烷基亞磺醯胺以形成 (H)來製備。 Embodiment 15 provides the method of any one of embodiments 13-14, wherein (H) is obtained by making ( R )-2-methylpropane-2-sulfinamide (G)

(G) with 1-(6,7-difluoro-1-methoxyisoquinolin-4-yl)ethan-1-one (F)

(F) is prepared by reacting to form the corresponding N-alkylenesulfinamide and treating the N-alkylenesulfinamide with a reducing agent to form (H) .

Embodiment 16 provides the method of embodiment 15, wherein (F) is reacted with (G) in the presence of a titanium(IV) alkoxide to form an N-alkylenesulfinamide.

Embodiment 17 provides the method of any of embodiments 15-16, wherein the reducing agent comprises a borohydride.

(E)與鹼金屬甲醇鹽反應來製備。 Embodiment 18 provides the method of any one of embodiments 15-17, wherein (F) is obtained by making 1-(1-chloro-6,7-difluoro-4-isoquinolinyl)ketene (E)

(E) Prepared by reaction with alkali metal methoxide.

(D)與氯化劑反應來製備。 Embodiment 19 provides the method of embodiment 18, wherein (E) is obtained by making 4-acetoxy-6,7-difluoro-2H-isoquinolin-1-one ( D)

(D) is prepared by reacting with a chlorinating agent.

Embodiment 20 provides the method of embodiment 19, wherein the chlorinating agent comprises _POCl3 .

Embodiment 21 provides the method of embodiment 20, wherein the molar ratio of _POCl3 : (D) is about 1.2:1.

Embodiment 22 provides the method of any of embodiments 19-21, wherein (D) is reacted with a chlorinating agent in the presence of a base.

Embodiment 23 provides the method of embodiment 22, wherein the base comprises triethylamine, diisopropylethylamine, and/or pyridine.

Embodiment 24 provides the method of any of embodiments 19-23, wherein (D) is contacted with the chlorinating agent at a temperature not exceeding about 35°C.

Embodiment 25 provides the method of embodiment 24, wherein the mixture comprising (D) and the chlorinating agent is maintained at a temperature not exceeding about 80°C.

(C)、

(C')與選自甲醯胺和三𠯤的至少一種試劑反應來製備。 Embodiment 26 provides the method of any one of embodiments 19-25, wherein (D) is obtained by making 6,7-difluoro-3-methyl-1H-isochromen-1-one (C) or methyl 4 ,5-Difluoro-2-(2-oxypropyl)benzoate (C')

(C) ,

(C') is prepared by reacting with at least one reagent selected from the group consisting of formamide and tris.

Embodiment 27 provides the method of embodiment 26, wherein (C) or (C') is reacted with at least one reagent in the presence of a base.

Embodiment 28 provides the method of embodiment 27, wherein the base comprises potassium carbonate, sodium carbonate, sodium methoxide, and/or potassium methoxide.

Embodiment 29 provides the method of any of embodiments 27-28, wherein (C) or (C') is reacted with at least one reagent in the presence of a solvent.

Embodiment 30 provides the method of embodiment 29, wherein the solvent comprises 2-methyltetrahydrofuran and/or methanol.

(B)與酸反應來製備。 Embodiment 31 provides the method of any one of embodiments 26-30, wherein (C) or (C') is obtained by making 4,5-difluoro-2-(2-oxypropyl)benzoic acid (B)

(B) is prepared by reacting with an acid.

Embodiment 32 provides the method of embodiment 31, wherein ( B) is reacted with an acid in the presence of a solvent.

Embodiment 33 provides the method of embodiment 32, wherein the solvent is dichloromethane (DCM) or methanol (MeOH).

(A)與乙醯丙酮反應來製備。 Embodiment 34 provides the method of embodiment 31, wherein (B) is obtained by subjecting 2-bromo-4,5-difluorobenzoic acid to (A)

(A) Prepared by reacting with acetone acetone.

Embodiment 35 provides the method of embodiment 34, wherein (A) is reacted with acetone in the presence of a Lewis acid.

Embodiment 36 provides the method of embodiment 35, wherein the Lewis acid comprises a cuprous (I) salt.

Embodiment 37 provides the method of any one of embodiments 34-36, wherein (A) is reacted with acetone in the presence of a base.

Embodiment 38 provides the method of embodiment 37, wherein the base comprises an alkali metal alkoxide.

Embodiment 39 provides ( R )-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxo-1,2-dihydroisoquinoline) -4-yl)ethyl)-1-methylurea (X) crystalline solid characterized by an X-ray diffraction pattern (XRDP) selected from: (a) Form 1, whose X-ray powder diffraction spectrum includes About: 2Θ values (in degrees) at 7.1, 8.1, 25.8, 26.1, and 26.5; (b) Form 2, whose X-ray powder diffraction spectrum includes about: 11.29, 12.02, 17.51, 21.84, 22.09, and 22.72 2Θ Values (in degrees); (c) Form 3, whose X-ray powder diffraction spectrum includes 2Θ values (in degrees) of approximately: 10.78, 11.28, 17.65, 19.10, and 26.45; (d) Form 4, which X-ray powder diffraction spectrum including about: 8.0, 10.8, 13.2, 25.2, 25.5, and 26.5 2Θ values (in degrees); (e) Form 5, whose X-ray powder diffraction spectrum includes about: 7.09, 8.06, 2Θ values (in degrees) of 15.57, 15.86, 25.72, 26.07, 26.42 and 26.50; (f) Form 6, whose X-ray powder diffraction spectrum includes 2Θ values of approximately: 5.2, 7.8, 10.1, 11.3, 13.8 and 25.7 Values (in degrees); (g) Form 7, whose X-ray powder diffraction spectrum includes 2-theta values (in degrees) of about: 9.0, 12.9, 13.5, 20.4, and 26.0; (h) Form 8, which The X-ray powder diffraction spectrum includes 2-theta values (in degrees) of about: 5.2, 7.8, and 13.9; and (i) Form 9, whose X-ray powder diffraction spectrum includes 2-theta values of about: 4.0, 7.2, and 8.1 ( in degrees); where XRDP is measured with a copper X-ray source.

Embodiment 40 provides the solid of embodiment 39 characterized by an X-ray diffraction pattern (XRDP) selected from the group consisting of: and The 2θ value of 32.6 (in degrees); (b) Form 2, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 8.26, 18.32, 19.58, 21.25, 25.19, 28.15, and 29.54; (c) Form 3, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 8.34, 12.49, 14.37, 18.65, 22.66, 23.20, and 24.03; (d) Form 4, whose X-ray powder diffraction spectrum includes 2Θ values (in degrees) of about: 5.1, 8.6, 9.7, 10.3, 12.5, 16.9, 18.4, 23.6, 25.9, and 30.1; (e) Form 5, whose X-ray powder diffraction spectrum includes 2-theta values (in degrees) of about: 9.58, 12.12, 13.17, 14.15, 26.77, and 27.67; (f) Form 6, whose X-ray powder diffraction spectrum includes 2Θ values (in degrees) of about: 7.4, 9.5, 10.9, 11.5, 12.7, 19.1, 22.5, 22.7, 26.0, 26.5, 26.7, and 27.9; (g) Form 7 having an X-ray powder diffraction spectrum comprising about: 4.0, 7.0, 7.6, 7.8, 9.6, 10.1, 10.7, 12.1, 14.7, 19.9, 20.7, 21.0, 22.9, 25.2, 26.4, 27.0, 27.8 and 28.8's 2theta value in degrees; (h) Form 8, whose X-ray powder diffraction spectrum includes 2Θ values (in degrees) of about: 7.4, 9.5, 10.1, 11.4, 11.6, 25.7, 26.9, and 28.1; and (i) Form 9, whose X-ray powder diffraction spectrum includes 2-theta values (in degrees) of about: 6.5, 9.6, 10.8, 11.1, 12.1, 13.2, 14.4, 15.4, 15.7, 19.1, 26.0, and 26.4.

Embodiment 41 provides the solid of any of Embodiments 39-40, characterized by an X-ray diffraction pattern (XRDP) selected from the group consisting of: (a) Form 1, the X-ray powder diffraction spectrum of which includes about: 2Theta values (in degrees) for 21.0, 21.4, 21.9, 22.9, 24.0, 24.3, 25.3, 25.8, 26.1, 26.5, 27.0, 27.9, 28.6, 30.5, and 32.6; (b) Form 2, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 12.2, 12.4, 14.27, 17.22, 18.00, 20.06, 24.72, 27.49, 31.74, 32.10, and 33.06; (c) Form 3, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 24.76, 25.27, 28.37, 30.52, and 32.67; (d) Form 4, the X-ray powder diffraction spectrum of which includes about: 23.0, 24.0, 24.3, 26.9, 27.3, 27.8, 28.4, 28.8, 29.5, 30.5, 31.3, 31.6, 32.1, 32.4, 32.9, 33.5, 33.9, 34.5, 35.4, 35.7, 36.1, 36.7, 38.0, 38.2, 39.4, 2θ values (in degrees) of 39.9 and 40.7; (e) Form 5 having an X-ray powder diffraction spectrum comprising about: 4.03, 6.45, 17.34, 19.27, 19.80, 20.79, 21.62, 22.13, 22.51, 23.53, 24.40, 27.94, 28.55, 29.24, 29.54, 30.38, 30.73 and The 2θ value of 32.71 in degrees; (f) Form 6, the X-ray powder diffraction spectrum of which includes about: 23.3, 24.0, 24.5, 25.0, 25.5, 27.3, 28.5, 29.7, 29.9, 30.6, 31.0, 31.6, 32.0, 32.3, 32.5, 33.1, 33.7, 34.3, 34.6, 34.9, 36.0, 36.6, 37.1, 38.7, 40.1, 2θ values (in degrees) for 40.7 and 41.2; (g) Form 7, the X-ray powder diffraction spectrum of which includes about: 22.6, 23.3, 24.1, 24.4, 27.2, 28.2, 28.4, 29.2, 30.0, 30.5, 30.7, 31.4, 31.8, 32.6, 32.9, 33.3, 33.7, 34.9, 35.3, 35.8, 36.6, 37.3, 37.5, 38.2, 38.9, 2θ values (in degrees) for 39.4, 40.1, 40.3 and 41.2; (h) Form 8 having an X-ray powder diffraction spectrum comprising about: 2Theta values (in degrees) for 23.2, 24.0, 25.0, 26.1, 26.5, 27.4, 30.0, 30.8, 31.8, 32.5, 33.0, 33.7, 34.9, 35.8, 36.1, 38.7, and 41.2; and (i) Form 9, whose X-ray powder diffraction spectrum includes 2Θ values (in degrees) of about: 16.7, 17.6, 22.7, 23.2, 24.1, 24.4, 27.3, 28.2, 28.8, 30.8, and 32.7.

Embodiment 42 provides the solid of embodiment 39(a), wherein the solid is obtained by crystallizing (X) in at least one solvent selected from methanol and dichloromethane.

Embodiment 43 provides the solid of embodiment 39(a) comprising dichloromethane.

Embodiment 44 provides the solid of embodiment 39(b), wherein the solid is prepared by mixing in a mixture selected from the group consisting of water, ethyl acetate, acetone/water mixture, 2-methyltetrahydrofuran, 2-propanol, n-heptane, toluene, acetone, Obtained by crystallizing (X) from at least one solvent of a water/tetrahydrofuran mixture and dichloromethane.

Embodiment 45 provides the solid of embodiment 44, wherein the ratio of water:tetrahydrofuran in the water/tetrahydrofuran mixture is about 1:1 (v/v).

Embodiment 46 provides the solid of embodiment 44, wherein the ratio of acetone:water in the acetone/water mixture is about 4:1 (v/v), and the solid is vacuum dried at room temperature for about 1 day.

Embodiment 47 provides the solid of embodiment 39 (c), wherein the solid is prepared by dissolving (X) in at least one solvent selected from the group consisting of acetone, dichloromethane, a dimethylformamide/tert-butyl methyl ether mixture, and tetrahydrofuran. to obtain crystals.

Embodiment 48 provides the solid of embodiment 47, wherein the ratio of dimethylformamide:tertiary butyl methyl ether in the dimethylformamide/tertiary butyl methyl ether mixture is about 1:10 (v/v).

Embodiment 49 provides the solid of embodiment 39 (d), wherein the solid is obtained by crystallizing (X) in ethanol.

Embodiment 50 provides the solid of embodiment 39(d) comprising ethanol.

Embodiment 51 provides the solid of embodiment 39 (e), wherein the solid is obtained by crystallizing (X) in ethyl acetate.

Embodiment 52 provides the solid of embodiment 39(e) comprising ethyl acetate.

Embodiment 53 provides the solid of embodiment 39 (f), wherein the solid is obtained by crystallizing (X) in methanol.

Embodiment 54 provides the solid of embodiment 39(f) comprising methanol.

Embodiment 55 provides the solid of embodiment 39 (g), wherein the solid is obtained by crystallizing (X) in an acetone/water mixture.

Embodiment 56 provides the solid of embodiment 39 (g), wherein the ratio of acetone:water in the acetone/water mixture is about 4:1 (v/v).

Embodiment 57 provides the solid of embodiment 39 (h), wherein the solid is obtained by crystallizing (X) in ethanol and vacuum drying at room temperature for about 4 days.

Embodiment 58 provides the solid of embodiment 39(i), wherein the solid is obtained by subjecting Form 1 of (X) to at least one cycle of a dynamic vapor sorption (DVS) experiment.

Embodiment 59 provides the solid of embodiment 39(i), wherein the solid is obtained by crystallizing (X) in water.

Embodiment 60 provides the solid of any of Embodiments 39-46, wherein at least one applies: The solid in (a) has a glass transition at about 127.9°C as characterized by differential scanning calorimetry (DSC) thermogram; The solid in (b) was characterized by DSC thermogram as having a single maximum at about 219°C.

Embodiment 61 provides a pharmaceutical composition comprising at least one pharmaceutically acceptable carrier and the solid of any of embodiments 39-60.

Embodiment 62 provides the pharmaceutical composition of Embodiment 61, which is a solid dosage form for oral administration.

Embodiment 63 provides the pharmaceutical composition of any one of Embodiments 61-62, which is part of a lozenge, dragee, drop, suppository, capsule, caplet, and/or gelcap.

Embodiment 64 provides the pharmaceutical composition of any one of Embodiments 61-63, further comprising at least one additional agent for the treatment of hepatitis virus infection.

Embodiment 65 provides the pharmaceutical composition of embodiment 64, wherein the at least one additional agent comprises at least one selected from the group consisting of: reverse transcriptase inhibitor; capsid inhibitor; cccDNA formation inhibitor; RNA destabilizer; targeting HBV Genomic oligonucleotides; immunostimulants such as checkpoint inhibitors (eg, PD-L1 inhibitors); GalNAc-siRNA conjugates targeting HBV gene transcripts; and therapeutic vaccines.

Embodiment 66 provides a method of treating or preventing Hepatitis B virus (HBV) infection in a subject, the method comprising administering to the subject a therapeutically effective amount of the solid of any one of Embodiments 39-60 or Embodiment 61 - The pharmaceutical composition of any one of 65.

Embodiment 67 provides a method of directly or indirectly inhibiting the expression and/or function of a viral capsid protein in a subject infected with hepatitis B virus, the method comprising administering to the subject a therapeutically effective amount of embodiments 39-60 The solid of any one or the pharmaceutical composition of any one of embodiments 61-65.

Embodiment 68 provides the method of any one of embodiments 65-67, wherein the subject is further administered at least one additional agent for the treatment of HBV infection.

Embodiment 69 provides the method of embodiment 68, wherein the solid or pharmaceutical composition and at least one additional pharmaceutical agent are co-formulated.

Embodiment 70 provides the method of any one of embodiments 65-69, wherein the subject is further infected with hepatitis D virus (HDV).

Embodiment 71 provides the method of any of embodiments 65-70, wherein the subject is a mammal.

Embodiment 72 provides the method of embodiment 71, wherein the mammal is a human.

The disclosure of each patent, patent application, and publication cited herein is hereby incorporated by reference in its entirety.

While the present invention has been disclosed with reference to specific embodiments, it will be apparent that other embodiments and variations of this invention can be devised by others skilled in the art without departing from the true spirit and scope of this invention. The claims are intended to be construed to include all such embodiments and equivalent modifications.

none

The following detailed description of illustrative embodiments of the invention will be better understood when read in conjunction with the accompanying drawings. For the purpose of illustrating the invention, there are shown in the drawings certain illustrative embodiments. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

Figure 1 is (R)-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxo-1,2-dihydroisoquinoline- Non-limiting illustration of polarized light microscopy (PLM) images of 4-yl)ethyl)-1-methylurea (X) Form 1.

Figures 2A-2K are non-limiting illustrations of hot stage micrographs of Form 1 of (X) at several temperatures: (A) 25°C; (B) 140°C; (C) 150°C; (D) 158 (E) 170°C; (F) 178°C; (G) 185°C; (H) 193°C; (I) 217°C; (J) 220°C; and (K) 222°C.

Figure 3 is a non-limiting illustration of the XRPD spectrum of Form 1 of (X) .

Figure 4 is a non-limiting view of overlapping XRPD spectra of (a) Form 1 of ( X ) at 25°C; (b) Form 2 of ( X ) at 25°C; and (c) Form 1 of ( X ) at 160°C illustrate.

Figure 5 is a non-limiting illustration of TG-FTIR analysis of Form 1 of (X) .

Figure 6 is a non-limiting illustration of DSC analysis of Form 1 of (X) .

Figure 7 is a non-limiting illustration of DVS analysis of Form 1 of (X) .

Figure 8 is a non-limiting illustration of DVS analysis of Form 1 of ( X ).

Figure 9 is a non-limiting illustration of overlapping XRPD spectra of ( X ) Form 1 (a) before DVS analysis; and (b) after DVS analysis (Form 9).

Figure 10 is a non-limiting illustration of TG-FTIR analysis of Form 1 (X) following DVS analysis (Form 9).

Figure 11 is a non-limiting illustration of a polarized light microscope (PLM) image of Form 2 of ( X ).

Figure 12 is a non-limiting illustration of the XRPD spectrum of Form 2 of (X) .

Figure 13 is a non-limiting non-limiting XRPD spectrum of (a) Form 1 of ( X ), a first sample of (b) Form 2 of ( X ), and (c) a second sample of Form 2 of (X) illustrate.

Figure 14 is a non-limiting illustration of TG-FTIR analysis of Form 2 of (X) .

Figure 15 is a non-limiting illustration of DSC analysis of Form 2 of (X) .

Figure 16 is a non-limiting illustration of DVS analysis of Form 2 of (X) .

Figure 17 is a non-limiting illustration of DVS analysis of Form 2 of (X) .

Figure 18 is a non-limiting illustration of the XRPD spectrum of Form 3 of (X) .

Figure 19 is a non-limiting illustration of TG-FTIR analysis of Form 3 of (X) .

Figure 20 is a non-limiting illustration of the XRPD spectrum of Form 4 of (X) .

Figure 21 is a non-limiting illustration of TG-FTIR analysis of Form 4 of (X) .

Figure 22 is a non-limiting illustration of the XRPD spectrum of Form 5 of (X) .

Figure 23 is a non-limiting illustration of TG-FTIR analysis of Form 5 of (X) .

Figure 24 is a non-limiting illustration of the XRPD spectrum of Form 6 of (X) .

Figure 25 is a non-limiting illustration of TG-FTIR analysis of Form 6 of (X) .

Figure 26 is a non-limiting illustration of the XRPD spectrum of Form 7 of (X) .

Figure 27 is a non-limiting illustration of TG-FTIR analysis of Form 7 of (X) .

Figure 28 is a non-limiting illustration of the XRPD spectrum of Form 8 of (X) .

Figure 29 is a non-limiting illustration of overlapping XRPD spectra of ( a ) Form 4 (X) and (b) Form 8 (X) .

Figure 30 is a non-limiting illustration of TG-FTIR analysis of Form 8 of (X) .

Figure 31 is a non-limiting illustration of the results of a competitive slurry experiment for (a) Form 2 of (X) ; (b) Form 3 of ( X ); and Form 2 including (X) and (X) Overlaid XRPD spectra of the product of a competitive slurry experiment of Form 3 in acetone at (c) rt; and (d) 50°C.

Figure 32 is (a) Form 1 of (X) ; (b) Form 9 after DVS of Form 1 of (X) ; (c) Form 2 of (X) ; and (d) Form 4 of (X) A non-limiting illustration of the overlapping XRPD spectra.

Figure 33 is a non-limiting view of overlapping XRPD spectra of (a) Form 1 of (X) ; (b) Form 2 of (X) ; (c) Form 5 of (X) ; and (d) Form 7 of (X) Sexual description.

Figure 34 is a collection from different solvents and recrystallization techniques comprising (a) water slurry; (b) 2-methyl THF slurry; (c) 2-propanol slurry; and (d) n-heptane slurry A non-limiting illustration of the overlapping XRPD spectra of several independent samples of Form 2 of (X) .

Figure 35 includes (a) Form 3 obtained from methanol slurry of Form 1 of ( X ); (b) Form 8 obtained from ethanol slurry of Form 1 of (X) and subsequently dried; (c) Form 8 obtained from toluene slurry and (d) a non-limiting illustration of the overlapping XRPD spectra of Form 2 obtained from an aqueous slurry.

Figure 36 is a non-limiting view of the overlapping XRPD spectra of (a) DVS-treated Form 1 of (X) ; and (b) a crystalline solid collected from an aqueous slurry of (X ) Form 1 heated at 40°C for 6 days Sexual description.

Claims (72)

A preparation of ( R )-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxygen-1,2-dihydroisoquinoline-4 - base)ethyl)-1-methylurea (X) , or a method for a salt or solvate thereof,

(X) , the method comprising making (3-cyano-4-fluorophenyl) phenylcarbamate (K) :

(K) with ( R )-6,7-difluoro-4-(1-(methylamino)ethyl)isoquinolin-1(2H)-one (J) :

(J) reacts to produce a first reaction system comprising (X) . The method of claim 1, wherein the reaction of the (K) with the (J) is carried out in the presence of a base. The method of claim 2, wherein the base comprises triethylamine, diisopropylethylamine and/or pyridine. The method of claim 1, wherein the reaction of the (K) with the (J) is carried out in a solvent comprising 2-methyltetrahydrofuran. The method of claim 4, wherein the purification of (X) comprises neutralizing at least a portion of the base in the first reaction system and forming a first solution of 2-methyltetrahydrofuran of (X) produced. The method of claim 5, wherein the purification of (X) further comprises exchanging at least a portion of the 2-methyltetrahydrofuran with ethyl acetate in the first solution, thereby forming a second reaction system. The method of claim 6, wherein at least a portion of the second reaction system is concentrated to form solid (X) . The method of claim 1, wherein the (K) is prepared by reacting 5-amino-2-fluorobenzonitrile with phenyl chloroformate. The method of claim 8, wherein the 5-amino-2-fluorobenzonitrile is reacted with the phenyl chloroformate in the presence of a base. The method of claim 9, wherein the base comprises triethylamine, diisopropylethylamine and/or pyridine. The method of claim 1, wherein the (J) is prepared by making ( R )-N-(( R )-1-(6,7-difluoro-1-methoxyisoquinolin-4-yl ) ethyl)-N,2-dimethylpropane-2-sulfinamide (I) reacts with acid to prepare:

(I) . The method of claim 11, wherein the acid comprises HCl in ether and/or acetyl chloride in methanol. The method of claim 11, wherein by methylating ( R )-N-(( R )-1-(6,7-difluoro-1-methoxyisoquinolin-4-yl)ethyl base)-2-methylpropane-2-sulfinamide (H) to prepare this (I) :

(H) . The method of claim 13, wherein the methylating agent comprises methyl iodide, methyl chloride, methyl trifluoromethanesulfonate and/or dimethyl sulfate. The method of claim 13, wherein the (H) is prepared by making ( R )-2-methylpropane-2-sulfinamide (G)

(G) with 1-(6,7-difluoro-1-methoxyisoquinolin-4-yl)ethan-1-one (F)

(F) Reaction to form the corresponding N-alkylenesulfinamide, and treatment of the N-alkylenesulfinamide with a reducing agent to form (H) Preparation. The method of claim 15, wherein the (F) is reacted with the (G) in the presence of a titanium(IV) alkoxide to form an N-alkylenesulfinamide. The method of claim 15, wherein the reducing agent comprises borohydride. The method of claim 15, wherein the (F) is prepared by reacting 1-(1-chloro-6,7-difluoro-4-isoquinolinyl)ketene (E) with an alkali metal methoxide :

(E) . The method of claim 18, wherein the (E) is prepared by adding 4-acetyl-6,7-difluoro-2H-isoquinolin-1-one ( D)

(D) Prepared by reaction with chlorinating agent. The method of claim 19, wherein the chlorinating agent comprises _POCl3 . The method of claim 20, wherein the _POCl3 : (D) molar ratio is about 1.2:1. The method of claim 19, wherein the (D) is reacted with the chlorinating agent in the presence of a base. The method of claim 22, wherein the base comprises triethylamine, diisopropylethylamine and/or pyridine. The method of claim 19, wherein the (D) is contacted with the chlorinating agent at a temperature not exceeding about 35°C. The method of claim 24, wherein the mixture comprising the (D) and the chlorinating agent is maintained at a temperature not exceeding about 80°C. The method of claim 19, wherein the ( D ) is prepared by making 6,7-difluoro-3-methyl-1H-isochromen-1-one ( C ) or 4,5-difluoro-2- Methyl (2-oxypropyl)benzoate ( C' )

( C ),

( C' ) is prepared by reacting with at least one reagent selected from the group consisting of formamide and tris. The method of claim 26, wherein the ( C ) or the ( C' ) is reacted with the at least one reagent in the presence of a base. The method of claim 27, wherein the base comprises potassium carbonate, sodium carbonate, sodium methoxide and/or potassium methoxide. The method of claim 26, wherein the ( C ) or the ( C' ) is reacted with the at least one reagent in the presence of a solvent. The method of claim 29, wherein the solvent comprises 2-methyltetrahydrofuran and/or methanol. The method of claim 26, wherein by adding 4,5-difluoro-2-(2-oxypropyl)benzoic acid ( B )

( B ) is reacted with an acid to prepare the ( C ) or the ( C' ). The method of claim 31, wherein the ( B ) is reacted with the acid in the presence of a solvent. The method of claim 32, wherein the solvent is dichloromethane (DCM) or methanol (MeOH). The method of claim 31, wherein by adding 2-bromo-4,5-difluorobenzoic acid ( A )

(A) is reacted with acetone acetone to prepare this ( B ). The method of claim 34, wherein the ( A ) is reacted with acetylacetone in the presence of Lewis acid. The method of claim 35, wherein the Lewis acid comprises a cuprous (I) salt. The method of claim 35, wherein the (A) is reacted with acetylacetone in the presence of a base. The method of claim 37, wherein the base comprises an alkali metal alkoxide. A ( R )-3-(3-cyano-4-fluorophenyl)-1-(1-(6,7-difluoro-1-oxygen-1,2-dihydroisoquinoline-4- (X) crystalline solid, characterized by X-ray diffraction patterns (XRDP) selected from the group consisting of: (a) Form 1, its X-ray powder diffraction Spectra include 2Θ values (in degrees) at about: 7.1, 8.1, 25.8, 26.1, and 26.5; (b) Form 2, whose X-ray powder diffraction spectrum includes about: 11.29, 12.02, 17.51, 21.84, 22.09, and 22.72 (c) Form 3, whose X-ray powder diffraction spectrum includes 2θ values (in degrees) of approximately: 10.78, 11.28, 17.65, 19.10, and 26.45; (d) Form 4 , whose X-ray powder diffraction spectrum includes 2θ values (in degrees) of about: 8.0, 10.8, 13.2, 25.2, 25.5, and 26.5; (e) Form 5, whose X-ray powder diffraction spectrum includes about: 7.09, 2Θ values (in degrees) of 8.06, 15.57, 15.86, 25.72, 26.07, 26.42 and 26.50; (f) Form 6, whose X-ray powder diffraction spectrum includes approximately: 5.2, 7.8, 10.1, 11.3, 13.8 and 25.7 (g) Form 7, whose X-ray powder diffraction spectrum includes 2 theta values (in degrees) of approximately: 9.0, 12.9, 13.5, 20.4, and 26.0; (h) Form 8 , whose X-ray powder diffraction spectrum includes 2-theta values (in degrees) of about: 5.2, 7.8, and 13.9; and (i) Form 9, whose X-ray powder diffraction spectrum includes about: 4.0, 7.2, and 8.1 2-theta Value (in degrees); where the XRDP was measured with a copper X-ray source. The solid of claim 39 characterized by a X-ray diffraction pattern (XRDP) selected from the group consisting of: (a) Form 1, whose X-ray powder diffraction spectrum further comprises about: 10.8, 16.5, 16.9, 17.3, 17.7, 18.3, 19.5, 21.0, 21.4, 21.9, 22.9, 24.0, 24.3, 25.3, 27.9, 28.6, 30.5 and the 2θ value of 32.6 in degrees; (b) Form 2, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 8.26, 18.32, 19.58, 21.25, 25.19, 28.15, and 29.54; (c) Form 3, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 8.34, 12.49, 14.37, 18.65, 22.66, 23.20, and 24.03; (d) Form 4, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 5.1, 8.6, 9.7, 10.3, 12.5, 16.9, 18.4, 23.6, 25.9, and 30.1; (e) Form 5, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 9.58, 12.12, 13.17, 14.15, 26.77, and 27.67; (f) Form 6, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 7.4, 9.5, 10.9, 11.5, 12.7, 19.1, 22.5, 22.7, 26.0, 26.5, 26.7, and 27.9; (g) Form 7, the X-ray powder diffraction spectrum of which further comprises about: and the 2θ value of 28.8 in degrees; (h) Form 8, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 7.4, 9.5, 10.1, 11.4, 11.6, 25.7, 26.9, and 28.1; and (i) Form 9, whose X-ray powder diffraction spectrum includes 2-theta values (in degrees) of about: 6.5, 9.6, 10.8, 11.1, 12.1, 13.2, 14.4, 15.4, 15.7, 19.1, 26.0, and 26.4. The solid of claim 40, characterized by a X-ray diffraction pattern (XRDP) selected from the group consisting of: (a) Form 1, the X-ray powder diffraction spectrum of which includes about: 2Theta values (in degrees) for 21.0, 21.4, 21.9, 22.9, 24.0, 24.3, 25.3, 25.8, 26.1, 26.5, 27.0, 27.9, 28.6, 30.5, and 32.6; (b) Form 2, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 12.2, 12.4, 14.27, 17.22, 18.00, 20.06, 24.72, 27.49, 31.74, 32.10, and 33.06; (c) Form 3, the X-ray powder diffraction spectrum of which further comprises 2Θ values (in degrees) of about: 24.76, 25.27, 28.37, 30.52, and 32.67; (d) Form 4, the X-ray powder diffraction spectrum of which includes about: 23.0, 24.0, 24.3, 26.9, 27.3, 27.8, 28.4, 28.8, 29.5, 30.5, 31.3, 31.6, 32.1, 32.4, 32.9, 33.5, 33.9, 34.5, 35.4, 35.7, 36.1, 36.7, 38.0, 38.2, 39.4, 2θ values (in degrees) of 39.9 and 40.7; (e) Form 5 having an X-ray powder diffraction spectrum comprising about: 4.03, 6.45, 17.34, 19.27, 19.80, 20.79, 21.62, 22.13, 22.51, 23.53, 24.40, 27.94, 28.55, 29.24, 29.54, 30.38, 30.73 and The 2θ value of 32.71 in degrees; (f) Form 6, the X-ray powder diffraction spectrum of which includes about: 23.3, 24.0, 24.5, 25.0, 25.5, 27.3, 28.5, 29.7, 29.9, 30.6, 31.0, 31.6, 32.0, 32.3, 32.5, 33.1, 33.7, 34.3, 34.6, 34.9, 36.0, 36.6, 37.1, 38.7, 40.1, 2θ values (in degrees) for 40.7 and 41.2; (g) Form 7, the X-ray powder diffraction spectrum of which includes about: 22.6, 23.3, 24.1, 24.4, 27.2, 28.2, 28.4, 29.2, 30.0, 30.5, 30.7, 31.4, 31.8, 32.6, 32.9, 33.3, 33.7, 34.9, 35.3, 35.8, 36.6, 37.3, 37.5, 38.2, 38.9, 2θ values (in degrees) for 39.4, 40.1, 40.3 and 41.2; (h) Form 8 having an X-ray powder diffraction spectrum comprising about: 2Theta values (in degrees) for 23.2, 24.0, 25.0, 26.1, 26.5, 27.4, 30.0, 30.8, 31.8, 32.5, 33.0, 33.7, 34.9, 35.8, 36.1, 38.7, and 41.2; and (i) Form 9, whose X-ray powder diffraction spectrum includes 2Θ values (in degrees) of about: 16.7, 17.6, 22.7, 23.2, 24.1, 24.4, 27.3, 28.2, 28.8, 30.8, and 32.7. The solid of claim 39(a), obtained by crystallizing (X) in at least one solvent selected from methanol and dichloromethane. The solid of claim 39(a), which comprises dichloromethane. A solid as claimed in claim 39(b), wherein the solid is prepared by mixing in water, ethyl acetate, acetone/water mixture, 2-methyltetrahydrofuran, 2-propanol, n-heptane, toluene, acetone, water/ The solid is obtained by crystallizing (X) from at least one solvent of a mixture of tetrahydrofuran and dichloromethane. The solid of claim 44, wherein the ratio of water:tetrahydrofuran in the water/tetrahydrofuran mixture is about 1:1 (v/v). The solid of claim 44, wherein the ratio of acetone:water in the acetone/water mixture is about 4:1 (v/v), and the solid is vacuum dried at room temperature for 1 day. The solid of claim 39(c), wherein (X) is obtained by crystallizing (X) in at least one solvent selected from the group consisting of acetone, dichloromethane, dimethylformamide/tert-butyl methyl ether mixtures and tetrahydrofuran to obtain the solid. The solid of claim 47, wherein the ratio of dimethylformamide: tertiary butyl methyl ether in the dimethylformamide/tertiary butyl methyl ether mixture is about 1:10 (v/v). The solid of claim 39(d) obtained by crystallizing (X) in ethanol. The solid of claim 39(d), which comprises ethanol. The solid of claim 39(e) obtained by crystallizing (X) in ethyl acetate. The solid of claim 39(e) comprising ethyl acetate. The solid of claim 39(f) obtained by crystallizing (X) in methanol. The solid of claim 39(f), which comprises methanol. The solid of claim 39(g) obtained by crystallizing (X) in an acetone/water mixture. The solid of claim 55, wherein the ratio of acetone:water in the acetone/water mixture is about 4:1 (v/v). The solid of claim 39(h) obtained by crystallizing (X) in ethanol and vacuum drying at room temperature for about 4 days. The solid of claim 39(i), wherein the solid is obtained by subjecting Form 1 of (X) to at least one cycle of a Dynamic Vapor Sorption (DVS) experiment. The solid of claim 39(i) obtained by crystallizing (X) in water. A solid as claimed in any one of claims 39 to 46, wherein at least one of: The solid in (a) has a glass transition at about 127.9°C as characterized by differential scanning calorimetry (DSC) thermogram; The solid in (b) was characterized by DSC thermogram as having a single maximum at about 219°C. A pharmaceutical composition comprising at least one pharmaceutically acceptable carrier and the solid of any one of claims 39 to 60. The pharmaceutical composition of claim 61, which is a solid dosage form for oral administration. The pharmaceutical composition of any one of claims 61 to 62, which is part of a lozenge, dragee, drop, suppository, capsule, caplet, and/or gelcap. The pharmaceutical composition of any one of claims 61 to 63, further comprising at least one additional agent for the treatment of hepatitis virus infection. The pharmaceutical composition of claim 64, wherein the at least one additional agent comprises at least one selected from the group consisting of: reverse transcriptase inhibitor; capsid inhibitor; cccDNA formation inhibitor; RNA destabilization HBV genome-targeting oligonucleotides; immunostimulants such as checkpoint inhibitors (eg, PD-L1 inhibitors); GalNAc-siRNA conjugates targeting HBV gene transcripts; and therapeutic vaccines . A method of treating, ameliorating and/or preventing hepatitis B virus (HBV) infection in a subject, the method comprising administering to the subject a therapeutically effective amount of any one of claims 39 to 60 A solid and/or a pharmaceutical composition as claimed in any one of claims 61 to 65. A method of directly or indirectly inhibiting the expression and/or function of a viral capsid protein in a subject infected with hepatitis B virus, the method comprising administering to the subject a therapeutically effective amount of any one of claims 39 to 60 The solid according to one item and/or the pharmaceutical composition according to any one of claims 61 to 65. The method of claim 66 or 67, wherein the subject is further administered at least one additional agent for the treatment of HBV infection. The method of claim 68, wherein the solid or pharmaceutical composition and the at least one additional pharmaceutical agent are co-formulated. The method of any one of claims 66 to 69, wherein the subject is further infected with hepatitis D virus (HDV). The method of any one of claims 66 to 70, wherein the subject is a mammal. The method of claim 71, wherein the mammal is a human.

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