New synthesis method

Abstract

The present disclosure generally relates to a process for the synthesis of AZD6482. In particular, the present disclosure also relates to a process for the synthesis of enantiomerically pure AZD6482. The present disclosure also relates to a process for the synthesis of enantiomerically pure intermediates. The present disclosure also relates to compounds prepared by the processes and the use of such compounds to prepare compositions and to treat disorders.

Description

[0001] New synthesis method

[0002] This application claims priority from Australian application no. 2023901984 filed 23

[0003] June 2023. The entire contents of this application are incorporated by reference herein.

[0004] Technical Field

[0005] This application relates to the synthesis of AZD6482 and related compounds, in particular to the synthesis of enantiomerically pure AZD6482, and to intermediates useful in the synthesis of enantiomerically pure AZD6482. The present application also relates to pharmaceutical compositions comprising AZD6482 and to methods of therapeutic and prophylactic treatments of thrombosis and related conditions in a subject using AZD6482. AZD6482 may be administered alone or in combination with a thrombolytic agent and / or an anticoagulant.

[0006] Background

[0007] The present invention relates to processes for making AZD6482, particularly enantiomerically pure AZD6482, and to intermediates useful therein. AZD6482 is also referred to as TBO-309, KIN-193, 2-[l-(7-methyl-2-(morpholin-4-yl)-4-oxo-4H-pyrido[l,2- a]pyrimidin-9-yl)ethylamino]benzoic acid or (J?)-2-((l-(7-methyl-2-morpholino-4-oxo-477- pyrido[l,2-a]pyrimidin-9-yl)ethyl)amino) benzoic acid and has the following formula (la):

[0008] Formula (la)

[0009] AZD6482 was described in W02004 / 016607, and is a selective inhibitor of phosphoinositide (PI) 3-kinase p. As described in W02009 / 093972 and WO2021 / 226677, AZD6482 has anti-platelet activity and may be useful in the treatment and / or prevention of thrombosis, for example in combination with a thrombolytic agent and / or an anticoagulant.

[0010] AZD6482 has a single asymmetric center, i.e. the compound exists as two enantiomers. It is desirable to obtain enantiomerically pure AZD6482. One reported process for preparing AZD6482 comes from the international PCT publication W02009 / 093972, and is shown in Scheme 1, below. According to the reported process, compound 3 can be synthesised by reacting malonyl dichloride with 2-amino-3-bromo-5-methylpyridine. Compound 3 is then reacted with morpholine in the presence of a base to produce compound 4. Compound 4 is then reacted with butyl vinyl ether in the presence of PdCh(dppf) / DIPEA to form the ketone 5. The ketone 5 is then reduced using NaBth to form the secondary alcohol 6. The alcohol 6 is reacted with phosphorous tribromide to form an alkyl halide, which is then reacted with 2- aminobenzoic acid to obtain compound 1. Scheme 1. Reagents, conditions, and yields', (i) DCM, malonyl dichloride, RT, 48h; (ii) THF, EtsN, N2, MsCl, 5 °C, 2.5h, then morpholine, 60 °C, 5h, then H2O, 65 °C, 3h, then 20 °C, overnight (94%); (iii) K2CO3, DMF / H2O, N2, DPPP / Pd(OAc)2, 90 °C, 48h (89%); (iv) MeOH, NaHBr. RT, Ih, then H2O (80%); and (v) CH2CI2, PBn, 40 °C, 2.5h, then ET3N, 2- aminobenzoic acid, 40 °C, overnight, then H2O, acetone and HC1.

[0011] In the above synthetic route both enantiomers are produced. In order to isolate the preferred enantiomer, compound 1 is methylated by reacting with Mel in the presence of DIPEA to form compound 7. The enantiomers of compound 7 are separated by chiral chromatography and the preferred enantiomer treated with NaOH to form the preferred enantiomer of compound 1, as shown in Scheme 2 below.

[0012] Scheme 2. Reagents, conditions, and yields, (vi) DMF, Mel, DIPEA, rt, overnight; (vii) Chiralpak AS HPLC column eluted with heptane / EtOH 20:80; (viii) THF and MeOH, NaOH and H2O, rt, 3 days, recrystallise from EtOH / TEO.

[0013] A drawback to the reported process is the inability to efficiently scale this chemistry to the quantities that are required for clinical investigation and manufacture for clinical development. CN104592222 describes a method for the synthesis of enantiomerically pure AZD6482. Despite CN104592222, there is a need to improve the synthesis of AZD6482, particularly the synthesis of AZD6482 with a high degree of the enantiomeric purity and / or in the quantities that are required for clinical investigation and manufacture for clinical development. There is also a need for a process that can be reproduced.

[0014] Summary

[0015] The present application relates to intermediate compounds useful in making AZD6482 and related compounds and the use of these compounds in the synthesis of enantiomerically pure AZD6482. The subject matter of the present disclosure is predicated in part on the development of an efficient and scalable process for preparing a compound of Formula (I), including an enantiomerically pure compound of Formula (la).

[0016] It will be appreciated that other aspects, embodiments, and examples of the compounds, pharmaceutical compositions, methods, or uses, are further described herein.

[0017] As described herein, there is provided an enantiomerically rich compound of formula (IV): wherein R1is selected from Ci-Ce alkyl or an optionally substituted C3-7 cycloalkyl. In some embodiments, R1is C1-6 alkyl. In some embodiments, R1is selected from -Me, Et, zz-Pr, z-Pr, zz-Bu and / -Bu. In some embodiments, R1is -CH3.

[0018] The compounds of formula (IV) can be formed with a high ratio of the S epimer relative to the R epimer. The ratio of SR epimer can be in the range of 95:5 to 100:0 and even 99:1 to 100:0 (i.e., at least 95%, or at least 99% S enantiomeric purity). The ability to obtain the compound of formula (IV) in a solid form is also advantageous.

[0019] Compounds of formula (IV) are thus useful intermediates in making AZD6482 and related compounds. Accordingly, in one aspect there is provided a process for preparing AZD6482 (e.g. enantiomerically pure AZD6482) comprising using a compound of formula (IV) as described herein as an intermediate or a starting product.

[0020] In some embodiments, the compound of formula (IV) is a compound of formula (IVa):

[0021]

[0022] In one aspect, there is provided an enantiomerically rich compound of formula (III): Formula (III), wherein R1and R2are independently selected from Ci-Ce alkyl or an optionally substituted C3-7 cycloalkyl. In some embodiments, R1and R2are independently Ci-Ce alkyl. In some embodiments, R1is C1-6 alkyl. In some embodiments, R1is selected from -Me, Et, n- Pr, z-Pr, z?-Bu and Z-Bu. In some embodiments, R1is -CH3. In some embodiments, R2is C1-6 alkyl. In some embodiments, R2is selected from -Me, Et, zz-Pr, z-Pr, zz-Bu and z-Bu. In some embodiments, R2is -CH3. In some embodiments, R1is CH3 and R2is CH3.

[0023] In some embodiments, the compound of formula (III) is a compound of formula (Illa):

[0024]

[0025] Formula (Illa).

[0026] The compounds of formula (III) can be formed with a high ratio of the R epimer relative to the S epimer. The ratio of R S epimer can be in the range of 95:5 to 100:0 and even 99: 1 to 100:0 (i.e., at least 95%, or at least 99% R enantiomeric purity). The ability to obtain the compound of formula (III) in a solid form is also advantageous.

[0027] Compounds of formula (III) are thus useful intermediates in making AZD6482 and related compounds. Accordingly, in one aspect there is provided a process for preparing AZD6482 (e.g. enantiomerically pure AZD6482) comprising using a compound of formula (III) as described herein as an intermediate or a starting product.

[0028] In one aspect, there is provided an enantiomerically rich compound of formula (II):

[0029] Formula (II), wherein R1and R2are independently selected from Ci-Ce alkyl or an optionally substituted C3-7 cycloalkyl. In some embodiments, R1and R2are independently C1-C6 alkyl.

[0030] In some embodiments, R1is C1-6 alkyl. In some embodiments, R1is selected from -Me, Et, n- Pr, z-Pr, z?-Bu and Z-Bu. In some embodiments, R1is -CH3. In some embodiments, R2is C1-6 alkyl. In some embodiments, R2is selected from -Me, Et, zz-Pr, z-Pr, zz-Bu and z-Bu. In some embodiments, R2is -CH3. In some embodiments, R1is CH3 and R2is CH3.

[0031] In some embodiments, the compound of formula (II) is a compound of formula (Ila): Formula (Ila).

[0032] The compounds of formula (II) can be formed with a high ratio of the R epimer relative to the S epimer. The ratio of R S epimer can be in the range of 95:5 to 100:0 and even 99: 1 to 100:0 (i.e., at least 95%, or at least 99% R enantiomeric purity).

[0033] Compounds of formula (II) are thus useful intermediates in making AZD6482 and related compounds. Accordingly, in one aspect there is provided a process for preparing AZD6482 (e.g. enantiomerically pure AZD6482) comprising using a compound of formula (II) as described herein as an intermediate or a starting product.

[0034] In some embodiments, there is provided a process for preparing an intermediate compound of Formula (IV) comprising: asymmetric reduction of the ketone of Formula (V) to form a compound of Formula

[0035] (IV). wherein R1is Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl. In some embodiments, R1is C1-6 alkyl. In some embodiments, R1is selected from -Me, Et, H-Pr, z-Pr, rz-Bu and Z-Bu. In some embodiments, R1is CH3.

[0036] In some embodiments, a ruthenium-catalyst is used for asymmetric reduction of the ketone. In some embodiments, the ruthenium-catalyst is RuCl[(S,S)-MsDpen]( -cymene).

[0037] In some embodiments, the asymmetric reduction uses a hydrogen source. In some embodiments, a hydrogen source for the asymmetric reduction comprises triethylamine and formic acid. In some embodiments, formic acid and triethylamine is used in a formic acid: tri ethylamine ratio of between about 5:1 and 1:5. In some embodiments, formic acid and triethylamine is used in a formic acid: triethylamine ratio of between about 1.2: 1 and 1:1.2, or about 1 : 1. In some embodiments, formic acid is added portionwise to a solution comprising the compound of Formula (V) and ruthenium-catalyst in solvent. In some embodiments, about 0.2 molar equivalents of formic acid are added every 30 minutes over three hours. In some embodiments, formic acid is added continuously, for example, over a period of 1 to 5 hours (e g. 2 to 3 hours) to a solution comprising the compound of Formula (V) and ruthenium- catalyst in solvent.

[0038] In some embodiments, the asymmetric reduction is performed in the presence of a solvent, wherein the solvent is di chloromethane.

[0039] In some embodiments, the process provides an enantiomerically pure compound of formula (IV). In some embodiments, the process provides a compound of formula (IV) having an enantiomeric purity of at least 95%, or at least 97%.

[0040] In some embodiments, there is also provided a process for producing a compound of Formula (I): the process comprising: a) converting a compound of Formula (V) to a compound of Formula (IV) according to the process as described herein; b) a Mitsunobu reaction of the compound of Formula (IV) with a compound of formula (VI) to form a compound of Formula (III): c) converting the compound of Formula (III) to form the compound of Formula (I)

[0041] R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl; and

[0042] R2is an optionally substituted C1-6 alkyl or an optionally substituted C3-7 cycloalkyl. In some embodiments, R1and R2are independently selected from Ci-Ce alkyl. In some embodiments, R1is C1-6 alkyl. In some embodiments, R1is selected from -Me, Et, zz-Pr, z-Pr, zz-Bu and t-Bu. In some embodiments, R1is -CH3. In some embodiments, R2is C1-6 alkyl. In some embodiments, R2is selected from -Me, Et, zz-Pr, z-Pr, zz-Bu and / -Bu. In some embodiments, R2is -CH . In some embodiments, R1is CH3 and R2is CH . In some embodiments, the Mitsunobu reaction is performed in the presence of triphenylphosphine (TPP) and an azodicarboxylate. In some embodiments, the azodicarboxylate is selected from diethyl azodicarboxylate (DEAD) or diisopropyl azodicarboxylate (DIAD).

[0043] In some embodiments, the Mitsunobu reaction is performed in the presence of a solvent, e.g. a non-polar or aprotic solvent. In some embodiments, the aprotic solvent or non-polar solvent is selected from dichloromethane (DCM), tetrahydrofuran (THF), methyltetrahydrofuran (MeTHF), acetonitrile (MeCN), A'-methylpyrrolidone (NMP), pyridine, toluene, hexanes, n-heptane, ethyl acetate (EtOAc), methylisopropyl ketone (MIPK), N,N- dimethylformamide (DMF), dimethylsulfoxide (DMSO), or any combinations thereof.

[0044] In some embodiments, the compound of formula (III) is isolated prior to use in step (c), wherein the isolation comprises: solvent exchange into THF; filtration to form a filtrate; solvent exchange of the filtrate into an alcohol to precipitate the compound of formula (III); and isolation of the precipitate. In some embodiments, the alcohol is selected from methanol, ethanol and isopropyl alcohol, or any combinations thereof. In some embodiments, wherein the alcohol is ethanol.

[0045] In some embodiments, the compound of Formula (VI) is prepared by a process comprising sulfonylation of an anthranilate of Formula (VII) using a nosylating agent:

[0046] Formula (VII) Formula (VI) wherein wherein, R2is Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl. In some embodiments, R2is Ci-6 alkyl. In some embodiments, R2is selected from -Me, Et, n-Pr, z-Pr, zz-Bu and Z-Bu. In some embodiments, R2is -CH3.

[0047] In some embodiments, the nosylating agent is 4-nosylchloride.

[0048] In some embodiments, the anthranilate of Formula (VII) is methyl anthranilate.

[0049] In some embodiments, the nosylating agent is provided in molar equivalents of between about 0.01 and 5, between about 0.05 and 4, or between about 0.1 and 2, or between about 0.5 to 1.5, or about 1.1 relative to the anthranilate. In some embodiments, a base reagent for sulfonylation of the anthranilate of Formula (VII) comprises pyridine. In some embodiments, a base reagent for sulfonylation of the anthranilate of Formula (VII) does not comprise DMAP. In some embodiments, a base reagent for sulfonylation of the anthranilate of Formula (VII) does not comprise tri ethylamine.

[0050] In some embodiments, the sulfonylation reaction is performed in the presence of a solvent and wherein the amount of solvent is present in volume equivalents (L), relative to the molar amount of the compound of the anthranilate of Formula (VII), of between about 1 and 15, between about 5 and 10, or between about 6 and 8. In some embodiments, the solvent is

[0051] DCM.

[0052] In some embodiments, step c) comprises: cl) de-nosylating the compound of Formula (III) to form a compound of Formula (II): c2) treating the compound of Formula (II) with a base to form a compound of Formula

[0053] (I):

[0054] In some embodiments, de-nosylating comprises reacting the compound of Formula (III) with thioglycolic acid in the presence of a base. In some embodiments, the base is tetramethylguanidine.

[0055] In some embodiments, the base used in step c2) is NaOH.

[0056] In some embodiments, the compounds are reacted in situ, without isolation, in the consequent reaction. In some embodiments, further comprising recrystallising the compound of Formula (I) using iso-propanol.

[0057] In some embodiments,

[0058] In some embodiments, the process provides a compound of formula (I) having an enantiomeric purity of at least 95%, or at least 97%.

[0059] In some embodiments, there is also provided an enantiomerically pure compound of Formula (IV) wherein Ri is C1-6 alkyl or an optionally substituted C3-7 cycloalkyl. In some embodiments, R1is Ci-Ce alkyl. In some embodiments, R1is selected from -Me, Et, «-Pr, i- Pr, n-Bu and Z-Bu. In some embodiments, R1is -CH3.

[0060] In some embodiments, there is also provided an enantiomerically pure compound of Formula (IVa):

[0061] Formula (IVa)

[0062] In some embodiments, there is also provided an enantiomerically pure compound of Formula (III):

[0063] Formula (III) wherein

[0064] R1and R2are independently Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl, and

[0065] In some embodiments, R1and R2are independently Ci-Ce alkyl. In some embodiments, R1is C1-6 alkyl. In some embodiments, R1is selected from -Me, Et, zz-Pr, z-Pr, zz-Bu and Z-Bu In some embodiments, R1is -CH3. In some embodiments, R2is C1-6 alkyl. In some embodiments, R2is selected from -Me, Et, zz-Pr, z-Pr, zz-Bu and Z-Bu. In some embodiments, R2is -CH3. In some embodiments, R1is CH3 and R2is CH3.

[0066] In some embodiments, there is also provided an enantiomerically pure compound of

[0067] Formula (Illa):

[0068]

[0069] Formula (Illa) wherein

[0070] In some embodiments, there is also provided a compound of Formula (I) prepared by one or more of the process described herein. In some embodiments, the compound is enantiomerically pure. In some embodiments, the compound has an enantiomeric purity of at least 95%, or at least 97%.

[0071] In some embodiments, there is also provided a process for preparing AZD6482 starting from a compound of Formula (IVa). In some embodiments, AZD6482 has an enantiomeric purity of at least 95%, or at least 97%.

[0072] In other aspects or embodiments, there is provided a compound of Formula (III), Formula (II), or Formula (I), prepared from any one or more processes as described by the above aspects, or any embodiments or examples thereof as described herein.

[0073] In other embodiments or examples, the process of preparing a compound of Formula (I) comprises the process of one or more of the above further aspects.

[0074] In some aspects, the present application provides enantiomerically pure AZD6482. In some embodiments, the enantiomeric purity is at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%.

[0075] Particular embodiments of each aspect are described throughout the specification, including in the dependent claims. The present invention is not to be limited in scope by the specific embodiments described herein, which are intended for the purpose of exemplification only. Functionally-equivalent products, compositions and methods are clearly within the scope of the invention as described herein.

[0076] Brief description of the drawings

[0077] The following figures form part of the present specification and are included to further demonstrate certain aspects of the present disclosure. The disclosure may be better understood by reference to one or more of these figures in combination with the detailed description of specific embodiments presented herein.

[0078] Figure 1 shows the X-ray powder diffraction pattern (XRPD) of an example compound of Formula (I), 2-[l-(7-methyl-2-(morpholin-4-yl)-4-oxo-4H-pyrido[l,2-a]pyrimidin-9 yl)ethylamino]benzoic acid.

[0079] Figure 2 shows the X-ray crystal structure of an example compound of Formula (I), 2-[l-(7- methyl-2-(morpholin-4-yl)-4-oxo-4H-pyrido[ 1 ,2-a]pyri midin-9 yl)ethylamino]benzoic acid.

[0080] Figure 3 shows the infrared spectrum of an example compound of Formula (I), 2-[l-(7-methyl- 2-(morpholin-4-yl)-4-oxo-4H-pyrido[ 1 ,2-rz]pyrimidin-9 yl)ethylamino]benzoic acid.

[0081] Figure 4 shows the Differential scanning calorimetry (DSC) of an example compound of Formula (I), 2-[l-(7-methyl-2-(morpholin-4-yl)-4-oxo-4H-pyrido[l,2-rz]pyrimidin-9- yl)ethylamino]benzoic acid. The DSC thermogram was obtained within the range 30 - 300 °C under inert atmosphere. The DSC thermogram shows one endothermic event with measured on-set temperature at 255.0 °C.

[0082] Figure 5 shows an example scheme for the synthesis of an example compound of Formula (I), 2-[l-(7-methyl-2-(morpholin-4-yl)-4-oxo-4H-pyrido[l,2-a]pyrimidin-9-yl)ethylamino] benzoic acid. Detailed description

[0083] General Definitions

[0084] The following definitions apply to the terms as used throughout this specification, unless otherwise limited in specific instances.

[0085] Unless specifically defined otherwise, all technical and scientific terms used herein shall be taken to have the same meaning as commonly understood by one of ordinary skill in the art (e.g., chemistry, biochemistry, medicinal chemistry, and the like).

[0086] As used herein, the term “and / or”, e.g., “X and / or Y”, shall be understood to mean either "X and Y" or "X or Y" and shall be taken to provide explicit support for both meanings or for either meaning, e.g., A and / or B includes the options i) A, ii) B, or iii) A and B.

[0087] As used herein, the term “about”, unless stated to the contrary, refers to + / - 20%, typically + / - 10%, typically + / - 5%, of the designated value. For the avoidance of doubt, it is to be understood that the term “about” includes a specific reference to the integer (e.g. “about 10” is to be understood as including an explicit reference to 10).

[0088] As used herein, the terms “a”, “an”, and “the” include both singular and plural aspects, unless the context clearly indicates otherwise.

[0089] Throughout this specification, unless specifically stated otherwise or the context requires otherwise, reference to a single step, composition of matter, group of steps or group of compositions of matter shall be taken to encompass one and a plurality (i.e. one or more) of those steps, compositions of matter, groups of steps or group of compositions of matter.

[0090] It is to be appreciated that certain features that are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any sub-combination.

[0091] Those skilled in the art will appreciate that the present disclosure is susceptible to variations and modifications other than those specifically described. It is to be understood that the disclosure includes all such variations and modifications. The disclosure also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any two or more of said steps or features.

[0092] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purpose of exemplification only. Functionally equivalent products, compositions and methods are clearly within the scope of the disclosure, as described herein.

[0093] Each feature of any particular aspect or embodiment or embodiment of the present disclosure may be applied mutatis mutandis to any other aspect or embodiment or embodiment of the present disclosure.

[0094] Throughout the present specification, various aspects and components of the invention can be presented in a range format. The range format is included for convenience and should not be interpreted as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range, unless specifically indicated. For example, description of a range such as from 1 to 5 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 5, from 3 to 5 etc., as well as individual and partial numbers within the recited range, for example, 1, 2, 3, 4, 5, 5.5 and 6, unless where integers are required or implicit from context. This applies regardless of the breadth of the disclosed range. Where specific values are required, these will be indicated in the specification.

[0095] Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps.

[0096] It will be clearly understood that, although a number of prior art publications are referred to herein, this reference does not constitute an admission that any of these documents form part of the common general knowledge in the art in the United States, Australia, or in any other country.

[0097] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. In case of conflict, the present specification, including definitions, will prevail. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0098] Salts may be formed in the case of embodiments of the compounds described herein, which contain a suitable acidic or basic group. Suitable salts of the compounds described herein include those formed with organic or inorganic acids or bases. Accordingly, it will be appreciated, that in referring to a compound by its formula (i.e., Formula (I)), reference is made to both the free-base / free-acid compound, and the corresponding salt thereof.

[0099] As used herein, the phrase “pharmaceutically acceptable salt” or a like term refers to pharmaceutically acceptable organic or inorganic salts. It will be appreciated that any reference to “salt” herein can include “pharmaceutically acceptable salts”. Exemplary acid addition salts include, but are not limited to, sulfate, citrate, acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate, phosphate, isonicotinate, lactate, salicylate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, / ?-toluenesulfonate, and pamoate (i.e., l,l'-methylene-bis-(2 -hydroxy-3 - naphthoate)) salts Exemplary base addition salts include, but are not limited to, ammonium salts, alkali metal salts, for example those of potassium and sodium, alkaline earth metal salts, for example those of calcium and magnesium, and salts with organic bases, for example dicyclohexylamine, A-methyl-D-glucamine, morpholine, thiomorpholine, piperidine, pyrrolidine, a mono-, di- or tri-lower alkylamine, for example ethyl-, tert-butyl-, diethyl-, diisopropyl-, triethyl-, tributyl- or dimethyl- propylamine, or a mono-, di- or trihydroxy lower alkylamine, for example mono-, di- or triethanolamine. A pharmaceutically acceptable salt may involve the inclusion of another molecule such as an acetate ion, a succinate ion or other counterion. The counterion may be any organic or inorganic moiety that stabilizes the charge on the parent compound. Furthermore, a pharmaceutically acceptable salt may have more than one charged atom in its structure. Instances where multiple charged atoms are part of the pharmaceutically acceptable salt can have multiple counter ions. Hence, a pharmaceutically acceptable salt can have one or more charged atoms and / or one or more counterions. It will also be appreciated that non-pharmaceutically acceptable salts also fall within the scope of the present disclosure since these may be useful as intermediates in the preparation of pharmaceutically acceptable salts or may be useful during storage or transport.

[0100] Those skilled in the art of organic and / or medicinal chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as “solvates”. For example, a complex with water is known as a “hydrate”. As used herein, the phrase “pharmaceutically acceptable solvate” or “solvate” refer to an association of one or more solvent molecules and a compound of the present disclosure. Examples of solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropyl alcohol, ethanol, methanol, dimethylsulfoxide, ethyl acetate, isopropyl acetate, acetic acid, and ethanolamine. It will be understood that the present disclosure encompasses solvated forms, including hydrates, of the compounds of Formula (I) and salts thereof.

[0101] The compounds of the present disclosure may contain one or more chiral (asymmetric) centers or the molecule as a whole may be chiral. Unless indicated otherwise, the individual stereoisomers (e.g., enantiomers and / or diastereoisomers) and mixtures of these are within the scope of the present disclosure.

[0102] As used herein, the term “stereoisomer” refers to compounds having the same molecular formula and sequence of bonded atoms (i.e., atom connectivity), though differ in the three- dimensional orientations of their atoms in space. As used herein, the term “enantiomers” refers to two compounds that are stereoisomers in that they are non-superimposable mirror images of one another. Relevant stereocenters may be denoted with (R)- or (^-configuration.

[0103] Those skilled in the art of organic and / or medicinal chemistry will appreciate that the compounds described herein (e.g. compounds of formula (I)) and salts thereof may be present in an amorphous form or in a crystalline form. Unless indicated otherwise, it will be understood that the present disclosure encompasses all forms and polymorphs of the compounds described herein (e.g. compounds of formula (I)) and salts thereof.

[0104] As used herein, the term “protecting group” has the meaning conventionally associated with it in organic synthesis / medicinal chemistry, i.e., a chemical group that selectively blocks one or more reactive sites in a multifunctional compound such that a chemical reaction can be carried out selectively on another unprotected reactive site and such that the group can readily be removed after the selective reaction is complete.

[0105] Implicit hydrogen atoms (such as the hydrogen atoms present on the pyrrole ring, etc.) are omitted from the formulae for clarity, but would be understood by the skilled person to be present.

[0106] As used herein, the term “alkyl” encompasses both straight-chain (i.e., linear) and branched-chain hydrocarbon groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, i-butyl, sec-butyl, pentyl, and hexyl groups. In one example, the alkyl group is of one to eight carbon atoms (i.e. Ci-salkyl). In one example, the alkyl group is of one to six carbon atoms (i.e. Ci-ealkyl). In one example, the Ci-ealkyl comprises methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, i-butyl and sec-butyl. As used herein, “cycloalkyl” means a fully saturated carbocyclic ring or ring system. In some embodiments, the cycloalkyl is one of 3 to 7 carbon atoms (i.e. C3-7 cycloalkyl). Examples include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0107] As used herein, the term “optionally substituted” refers to a group being unsubstituted or substituted as described herein. For the avoidance of doubt, it is to be understood that the term “optionally substituted” includes a specific reference to the unsubstituted group (e g. “optionally substituted Cue alkyl” is to be understood as including an explicit reference to “unsubstituted C1-6 alkyl”).

[0108] As used herein, the term “unsubstituted” refers to a group that does not have any further groups attached thereto or substituted therefore.

[0109] As used herein, a substituted group is derived from the unsubstituted parent group in which there has been an exchange of one or more hydrogen atoms for another atom or group (i.e., substituent). Unless otherwise indicated, when a group is deemed to be “substituted,” it is meant that the group is substituted with one or more substituents independently selected from Ci-Ce alkyl, C2-C6 alkenyl, C2-C6 alkynyl, Ci-Ce heteroalkyl, C3-C7 carbocyclyl (optionally substituted with halo, Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce haloalkyl, and Ci-Ce haloalkoxy), C3- Cv-carbocyclyl-Ci-Ce-alkyl (optionally substituted with halo, Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce haloalkyl, and Ci-Ce haloalkoxy), 5-10 membered heterocyclyl (optionally substituted with halo, Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce haloalkyl, and Ci-Ce haloalkoxy), 5-10 membered heterocyclyl-Ci-Ce-alkyl (optionally substituted with halo, Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce haloalkyl, and Ci-Ce haloalkoxy), aryl (optionally substituted with halo, Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce haloalkyl, and Ci-Ce haloalkoxy), aryl(Ci-Ce)alkyl (optionally substituted with halo, Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce haloalkyl, and Ci-Ce haloalkoxy), 5-10 membered heteroaryl (optionally substituted with halo, Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce haloalkyl, and Ci-Ce haloalkoxy), 5-10 membered heteroaryl(Ci-Ce)alkyl (optionally substituted with halo, Ci-Ce alkyl, Ci-Ce alkoxy, Ci-Ce haloalkyl, and Ci-Ce haloalkoxy), halo, cyano, hydroxy, Ci- Ce alkoxy, Ci-Ce alkoxy(Ci-Ce)alkyl (i.e., ether), aryloxy, sulfhydryl (mercapto), halo(Ci- Ce)alkyl (e.g., -CF3), halo(Ci-Ce)alkoxy (e.g., -OCF3), Ci-Ce alkylthio, arylthio, amino, amino(Ci-Ce)alkyl, nitro, O-carbamyl, A-carbamyl, (9-thiocarbamyl, A-thiocarbamyl, C- amido, A-amido, 5-sulfonamido, A-sulfonamido, C-carboxy, O-carboxy, acyl, cyanato, isocyanato, thiocyanato, isothiocyanate, sulfinyl, sulfonyl, and oxo (=0). Wherever a group is described as “optionally substituted” that group can be substituted with the above substituents. As used herein, the term “saturated” refers to a group where all available valence bonds of the backbone atoms are attached to other atoms. Representative examples of saturated groups include, but are not limited to, butyl, cyclohexyl, piperidine, and the like.

[0110] As used herein, the term “unsaturated” refers to a group where at least one valence bond of two adjacent backbone atoms is not attached to other atoms. Representative examples include, but are not limited to, alkenes (e g., -CH2-CH2CH=CH), phenyl, pyrrole, and the like.

[0111] As used herein, the term “optionally substituted” refers to a group being unsubstituted or substituted as described herein.

[0112] The term "selective PI 3-kinase P inhibitor" as used herein refers to a compound that inhibits PI 3-kinase at least >10-fold, preferably >20-fold, more preferably >30-fold more effectively than other isoforms of the PI 3-kinase family. A "selective PI 3-kinase P inhibitor" compound is understood to be more selective for PI 3-kinase P than compounds conventionally and generally designated PI 3-kinase inhibitors such as LY294002 or wortmannin. Compounds of any type that selectively inhibit PI 3-kinase P expression or activity can be used as selective PI 3-kinase P inhibitors in the methods of the present invention.

[0113] The terms "chiral purity" and "enantiomeric purity" are used interchangeably throughout the specification and refer to a measure of the purity of a substance (enantiomer) with the undesired enantiomer being the impurity. Throughout this specification, the chiral purity or enantiomeric purity of products is expressed as e.r. or relative area of the desired enantiomer by chiral HPLC analysis (%-area) (these terms are used interchangeably). A HPLC chiral purity of 50 %-area indicates a racemic mixture. When enantiomeric purity is measured by HPLC, an enantiomeric purity of 98% indicates a 98% area under the curve (AUC) for the desired enantiomer compared to 2% AUC for the undesired enantiomer. As described herein, an enantiomerically pure compound has an enantiomeric purity of at least 90% or greater, about 91% or greater, about 92% or greater, about 93% or greater, about 94% or greater, about 95% or greater, or about 96% or greater, or about 97% or greater, or about 98% or greater, or about 99% or greater, or even about 99.9% or greater. In some embodiments, the product has an enantiomeric excess of at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%.

[0114] The term "enantiomeric excess" or “ee” refers to the absolute difference between the mole fraction of each enantiomers as further defined below. A racemic mixture has an ee of 0%, while a single completely pure enantiomer has an ee of 100%. In some embodiments, ee can be expressed as a percentage. All documents cited or referenced herein, and all documents cited or referenced in herein cited documents, together with any manufacturer’s instructions, descriptions, product specifications, and product sheets for any products mentioned herein or in any document incorporated by reference herein, are hereby incorporated herein by reference in their entirety.

[0115] Process for preparing AZD6482

[0116] The subject matter of the present disclosure is predicated in part on the discovery of an efficient and scalable process for preparing AZD6482, including enantiomerically pure AZD6482. Scheme 3, below, provides an example of an efficient and / or scalable process for

[0117] Scheme 3. Exemplary process for the preparation of AZD6482.

[0118] The above process is described further below in relation to each of the steps of the process. Each step may provide its own independent process aspect, embodiment, or example for preparing an intermediate or compound per se, or may provide a further embodiment or example to another process aspect or embodiment as described herein. Each intermediate or prepared compound of each step may also provide its own independent aspect, embodiment, or example, in relation to compounds, compositions, and / or processes thereof.

[0119] As will be appreciated by the person skilled in the art, the one or more of the processes described herein will be performed in a suitable solvent. In some embodiments, the reactions may be performed in a solvent, such as a non-polar solvent, polar protic solvent, or polar aprotic solvent. Examples of non-polar solvents include, but are not limited to, hydrocarbons and aromatic hydrocarbons and combinations thereof. Examples of hydrocarbons include pentane, hexane, and heptane. Examples of aromatics include benzene, toluene, and xylene. Examples of polar protic solvents include, but are not limited to, alcohols, glycols, and any combinations thereof. Examples of alcohols include, but are not limited to, methanol (MeOH), ethanol (EtOH), 1 -propanol, isopropyl alcohol (2 -propanol, z-PrOH or IP A), 1 -butanol, 2-butanol, t- butanol (7-BuOH), 1 -pentanol, 3 -methyl- 1 -butanol, 2-methyl-l -propanol, and combinations thereof. Examples of glycols include, but are not limited to, ethylene glycol. Examples of polar aprotic solvents include, but are not limited to, halogenated hydrocarbons, ketones, nitriles, esters, carbonate esters, ethers, sulfoxides, sulfones, amides, nitroalkanes, pyrrolidines, pyridines, and combinations thereof. Examples of ketones include, but are not limited to, acetone, methylethyl ketone (MEK), methylbutyl ketone (MBK), methylisobutyl ketone (MIBK), methylisopropyl ketone (MIPK), and combinations thereof. Examples of nitriles include, but are not limited to, acetonitrile (MeCN). Examples of esters include, but are not limited to, ethyl formate, methyl acetate (MeOAc), ethyl acetate (EtOAc), propyl acetate, isopropyl acetate (z-PAC), zz-butyl acetate, isobutyl acetate, and combinations thereof. Examples of carbonate esters include, but are not limited to, dimethyl carbonate (DMC), propylene carbonate (PC), and combinations thereof. Examples of polar and non-polar ethers include, but are not limited to, mcthyl-Ze / 7-butyl ether (MTBE), diethyl ether, 1,4-di oxane, 2- m ethoxy ethanol, 2-ethoxy ethanol, dimethoxyethane (DME or monoglyme), 1,1- dimethoxymethane, 2,2-dimethoxypropane, 1,1 -di ethoxypropane, isopropyl ether, petroleum ether, cyclopentyl methyl ether (CPME), anisole (methoxybenzene), methyltetrahydrofuran (MeTHF), tetrahydrofuran (THF), and combinations thereof. Examples of sulfoxides include, but are not limited to, dimethylsulfoxide (DMSO). Examples of sulfones include, but are not limited to, sulfolane. Examples of amides include, but are not limited to, formamide, A,A- dimethylacetamide, A-di methyl form am ide (DMF), and combinations thereof. Examples of nitroalkanes include, but are not limited to, nitromethane. Examples of pyrrolidines include, but are not limited to, A-methylpyrrolidone (NMP). Examples of pyridines include, but are not limited to, pyridine. Examples of polar and non-polar halogenated hydrocarbons, such as chlorocarbons, include, but are not limited to, di chloromethane (DCM), chloroform, 1,2- di chloroethane, 1,1,1 -tri chloroethane, 1,1 -di chloroethene, 1,2-di chloroethene, and combinations thereof. Examples of hydrocarbons include, but are not limited to, hexanes and n-heptane and combinations thereof.

[0120] In some embodiments, the solvent may be anhydrous. For example, the amount of water in the solvent may be less than about (in ppm) 100, 75, 50, 25, 10, 5, 1, 0.1, or 0.01. In some embodiments, the solvent is present in volumes (L) of greater than about 1, 10, 50, 75, 100, 250, 500, 750, 1000, 2000, 3000, 4000, or 5000. In some embodiments, the solvent is present in volumes (L) of less than about 10000, 5000, 4000, 3000, 2000, 1000, 750, 500, 250, or 100. The solvent may be in a range provided by any two or more of the upper and / or lower amounts, for example between about 1 and 10000, between about 100 and 5000, or between about 500 and 2000. These volumes may relate to a single batch reaction system. It will be appreciated that multiple batch reactions may be combined.

[0121] Synthesis of a Compound of Formula (V) The compound of Formula (V):

[0122] Formula (V), wherein R1is an optionally substituted C1-6 alkyl or an optionally substituted C3-7 cycloalkyl, may be prepared as process described in W02004 / 016607 and / or W02009 / 093972.

[0123] In some embodiments, R1is a Ci-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R1is CH3. In some embodiments, the compound of Formula (V) is a compound of Formula (Va):

[0124] Formula (Va).

[0125] Synthesis of a Compound of Formula The present disclosure provides a process for preparing an intermediate compound of

[0126] Formula (IV):

[0127] Formula (IV), wherein R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0128] In some embodiments, R1is a C1-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R1is CH3.

[0129] In some embodiments, the present disclosure provides a process for preparing an enantiomerically pure intermediate compound of Formula (IV). In some embodiments, the process for preparing an intermediate compound of formula

[0130] (IV) comprising converting a compound of formula (V) to a compound of formula (IV).

[0131] wherein R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0132] In some embodiments, R1is a C1-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, z-propyl, / / -butyl and / -butyl. In some embodiments, R1is CHs. In some embodiments, the process comprises asymmetric reduction of the ketone of formula (V) to form a compound of formula (IV). In some embodiments, the intermediate compound of formula (IV) is an enantiomerically pure intermediate compound of (IV). For example, in some embodiments, the process provides a compound of formula (IV) having an enantiomer purity of at least 90%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% S.

[0133] In some embodiments, the compound of formula (V) is a compound of Formula (Va):

[0134] Formula (Va) and the compound of formula (IV) is a compound of Formula (IVa):

[0135] In some embodiments, the ketone of Formula (V) is reduced asymmetrically under ruthenium-catalyzed transfer hydrogenation conditions to provide the alcohol of Formula (IV), preferrably in high enantiomeric purity. In some embodiments, the asymmetric reduction reaction is performed in the presence of a ruthenium-catalyst. Any suitable ruthenium-catalyst may be used. In some embodiments, the ruthenium-catalyst includes (but is not limited to) Ru( / ?-cymene)(5,S-MsDPEN)Cl, Ru(mesitylene)(5,5-MsDPEN)Cl, Ru(7?,A-teth-T sDPEN)Cl or Ru( / ?-cymene)( / < / ?-FsDPEN)Cl. In some embodiments, the ruthenium-catalyst has a ( .5)-ligand, for example Ru(p-cymene)(5,5-MsDPEN)Cl or Ru(Mesitylene)(S,5-MsDPEN)Cl . In some embodiments, the ruthenium-catalyst is RuCl[( , )-MsDpen]( -cymene) (available from FountainB ridge Ltd., UK; Strem Ltd., currently Ascensus Specialities). Advantages associated with the use of RuCl[(S,S)-MsDpen](p-cymene) include availability and cost.

[0136] In some embodiments, the catalyst may have an enantiomeric excess (ee) of at least 95%, 96%, 97%, 98%, 99% or 99.3%. In some embodiments, the catalyst may have an ee of at least 98%.

[0137] In some embodiments, the concentration of catalyst in the reaction solution is greater than about 0.001, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, or 4 mol%, relative to the amount of the ketone. In some embodiments, the concentration of catalyst in the reaction solution is less than about 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, or 0.01 mol %, relative to the amount of the ketone. The catalyst may be in a concentration range provided by any two of these upper and / or lower values, for example between about 0.001 and 5, between about 0.005 and 2, or between about 0.01 and 1. In some embodiments, the concentration of catalyst in the reaction solution is 0.5 mol%. In some embodiments, the concentration of catalyst in the reaction solution is 1 mol%

[0138] As will be appreciated by the person skilled in the art, ruthenium-catalysed transfer hydrogenation is performed in the presence of a hydrogen source (i.e. hydrogen donor). Any suitable hydrogen source may be used. The hydrogen source may be provided in the reaction in any amount suitable so as to effect the asymmetric reduction reaction. In some embodiments, the hydrogen source is provided in molar equivalents, relative to the molar amount of the ketone of Formula (V), of less than about 5, 4, 3, 2, 1.5, 1.2, 1.0 or 0.5. In some embodiments, the hydrogen source is provided in molar equivalents, relative to the molar amount of the ketone of Formula (V), of greater than about 0.01, 0.05, 0.1, 0.3, 0.5, 1, 1.5, 2, or 3. The hydrogen source may be in a range provided by any two of these upper and / or lower ranges, for example between about 0.01 and 5, between about 0.05 and 4, or between about 0.1 and 2, or between about 0.5 to 1.5.

[0139] In some embodiments, the hydrogen source comprises ethanol, isopropanol or a reagent such as triethylammonium formate. As would be understand by the person skilled in the hydrogen source may also function as the solvent (e.g. isopropanol or ethanol). In some embodiments, the hydrogen source comprises isopropanol. In some embodiments, the hydrogen source comprises formic acid. In some embodiments, the hydrogen source comprises triethylamine and formic acid. In some embodiments, the molar ratio of formic acid: triethylamine is between 5: 1 and 1 :5, orbetween 3:l and 1:3, orbetween2: l and 1 :3, or between 1: 1 and 1 :2. In some embodiments, the molar ratio of formic acid: triethylamine is between 1.5: 1 and 1 :1.5, or between 1.2: 1 and 1:1.2 or about 1.2:1. In some embodiments, the molar ratio of formic acid: tri ethylamine is about 1.2:1. In some embodiments, the formic acid is provided in molar equivalents of between about 0.5 and 5, between about 0.8 and 3, between about 1 and 2 or between about 1.0 and 1.5, or about 1.2, relative to the ketone or formula (V). In some embodiments, the triethylamine is provided in molar equivalents of between about 0.5 and 5, between about 0.8 and 3, between about 0.9 and 2 or between about 1.0 and 1.5, or about 1.0, relative to the ketone or formula (V).

[0140] In some embodiments, the reaction mixture further comprises a suitable base. For example, in some embodiments, the base comprises potassium / -butoxide, sodium / -butoxide or lithium / -butoxide. In some embodiments, the concentration of base in the reaction solution is greater than about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 5, 7, 8, 9 or 10 mol%, relative to the amount of the ketone. In some embodiments, the concentration of base in the reaction solution is less than about 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1 mol %, relative to the amount of the ketone. The base may be in a concentration range provided by any two of these upper and / or lower values, for example between about 1 and 10, between about 3 and 7, or between about 4 and 6 mol%. In some embodiments, the concentration of base in the reaction solution is 5 mol%.

[0141] In some embodiments, the present inventors have found that the asymmetric reduction reaction as described herein results in the formation of the desired alcohol as well as an impurity that could not be easily removed by recrystallization. The present inventors have found that excess formic acid not only promotes impurity formation (e.g. formation of the side-product eluting from HPLC at RRT (relative retention time) 1.18), but also reduces the reaction rate. Without wishing to be bound by theory, it is thought that the improved results are obtained when the formic acid: triethylamine molar ratio is kept close to 1 :1 (e.g. 1.2: 1). This is in contrast to classic reaction conditions which often employ the azeotropic F / T 5:2 (mol / mol) mixture. By keeping the ratio close to 1 and avoiding an excess of formic acid, it is thought that the level of impurity is reduced.

[0142] In some embodiments, the asymmetric reduction reaction is performed in the presence of a solvent. In some embodiments, the solvent is a aprotic solvent or a non-polar. In some embodiments, the aprotic solvent or non-polar solvent is selected from hydrocarbons, halogenated hydrocarbons, aromatic hydrocarbons, ketones, nitriles, esters, carbonate esters, ethers, sulfoxides, sulfones, amides, nitroalkanes, pyrrolidines, or any combinations thereof. In some embodiments, the aprotic solvent or non-polar solvent is selected from di chloromethane (DCM), tetrahydrofuran (THF), methyltetrahydrofuran (MeTHF), acetonitrile (MeCN), N- methylpyrrolidone (NMP), pyridine, toluene, hexanes, n-heptane, ethyl acetate (EtOAc), methylisopropyl ketone (MIPK), 7V, V-di methyl form am ide (DMF), dimethylsulfoxide (DMSO), or any combinations thereof. In some embodiments, the solvent is dichloromethane (DCM). In some embodiments, the amount of solvent is present in volume equivalents (L), relative to the molar amount of the ketone of Formula (V), of between about 1 and 40, 1 and 30, between about 2 and 20, or between about 5 and 20, or between about 8 and 15, or about 10. In some embodiments, the solvent is a polar protic solvent. In some embodiments, the solvent is also the hydrogen source, for example, isopropanol or ethanol. In some embodiments, the solvent is isopropanol. In some embodiments, the solvent comprises isopropanol and dichloromethane.

[0143] In some embodiments, the concentration of hydrogen source in the reaction solution (mol / L) is greater than about 0.001, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, or 4. In some embodiments, the concentration of hydrogen source in the reaction solution (mol / L) is less than about 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, or 0.01. The hydrogen source may be in a concentration range provided by any two of these upper and / or lower values, for example between about 0.001 and 5, between about 0.005 and 2, or between about 0.01 and 1.

[0144] In some embodiments, the concentration of the compound of Formula (V) in the reaction solution (mol / L) is greater than about 0.001, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, or 4. In some embodiments, the concentration of the compound of Formula (V) in the reaction solution (mol / L) is less than about 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, or O.Ol. The compound of Formula (V) may be in a concentration range provided by any two of these upper and / or lower values, for example between about 0.001 and 5, between about 0.005 and 2, or between about 0.01 and 1.

[0145] In some embodiments, the compound of Formula (V) is dissolved in solvent and filtered prior to addition of the hydrogen source and catalyst. In some embodiments the solution containing the compound of Formula (V) is clear-filtered, such as on a fritted glass filter. The fritted glass filter may have an suitable porosity provided undissolved materials are retained. In one example, the porosity of the fritted glass filter is 3. Optionally, any undissolved compound of Formula (V) on the filter may be dissolved using additional solvent and filtered again. The clear filtrates are then optionally combined before addition of the hydrogen source and catalyst as defined herein. In some embodiments, the compound of Formula (V) recrystallised, for example using DCM, prior to addition of the hydrogen source and catalyst.

[0146] In some embodiments, the hydrogen source is added portion wise to a solution comprising the compound of Formula (V) and ruthenium-catalyst in solvent, e.g. the hydrogen source may be added as 2, 3, 4, 5, 6, 7, 8, 9, 10 or more portions. In some embodiments, the hydrogen source may be added in 6 portions. In some embodiments, the formic acid is added portionwise to a solution comprising the compound of Formula (V) and ruthenium-catalyst in solvent. In some embodiments, the solution further comprises triethylamine. Each portion may contain, for example, 0.1, 0.2, 0.3, 0.4 or 0.5 molar equivalents of formic acid, relative to the amount of the ketone. In some embodiments, each portion comprises about 0.2 molar equivalents of formic acid. In some embodiments, formic acid is added portionwise over a period of 1 or more hours, 2 or more hours or 3 or more hours. In some embodiments, a portion is added every 10 minutes, every 15 minutes, every 30 minutes, every 45 minutes or every 60 minutes. In some embodiments, a portion is added every 30 minutes. In some embodiments, 0.2 molar equivalents of formic acid is added every 30 minutes over three hours. Alternatively, the hydrogen source may be added in a slow stream (i.e. continuously), for example, over a period of several hours. In some embodiments, the hydrogen source is added over 1 hour, over 2 hours or over 3 or more hours. Without wishing to be bound by theory, it is thought addition of the hydrogen source (e.g. formic acid) portionwise or as a slow stream over a period of several hours ensures a faster reaction and / or limits side-product formation. Additional advantages of portion wise addition include that it offers good reproducibility between different scale batches and allows the reaction to reach almost full substrate conversion, for example, within 3 hours. The person skilled in the art will appreciate that it may be necessary to apply heat to facilitate the reaction. The amount of heat required may depend upon the solvent in which the reaction is performed in, as discussed above. In some embodiments, the mixture was heated to reflux. In some embodiments, the asymmetric reduction reaction is heated to between about 30 °C and 100 °C, about 40 °C and 80 °C, about 50 °C and 70 °C, or about 55 °C and 65 °C. In one example, the asymmetric reduction reaction is heated to between about 25 °C and 50 °C. In one example, the asymmetric reduction reaction is performed in DCM and heated to between about 30 °C and 45 °C. In one example, the asymmetric reduction reaction is performed in DCM and heated to about 30 °C. In one example, the asymmetric reduction reaction is performed in DCM and heated to about 45 °C. In some embodiments, the mixture is held at reflux for sufficient time for the reaction to go to completion. In some embodiments, the mixture is held at reflux for 0.5, 1, 1.5, 2, 3, 4, 5, 6, 9, 12, 15, 18, 21 or 24 hours or more. In some embodiments, the mixture is held at reflux for 60 minutes. In some embodiments, the mixture is held at reflux for 120 minutes. It was found that longer reflux times reduced the amount of impurity observed at RRT1.18, however this came with lower isolated product yield.

[0147] In some embodiments, the compound of Formula (IV) may be isolated once the reaction has completed. The compound of Formula (IV) may be isolated according to any suitable means as would be understood by the person skilled in the art. For example, isolation of the compound of Formula (IV) may be achieved by filtration, chromatography (e g. reverse-phase or normal phase column chromatography), extraction, or recrystallization. In some embodiments, the reaction mixture may be cooled and the product isolated by filtration. In some embodiments, the reaction mixture may be cooled to about 0 °C and then held at that temperature for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 4, 15, 16, 17, 18, 19 or 20 hours before the product is isolated by filtration. In some embodiments, the reaction mixture may be cooled to about 0 °C and then held at that temperature for at least 1 hour. In some embodiments, the reaction mixture may be cooled to about 0 °C and then held at that temperature for at least 10 hours. In some embodiments, the reaction mixture may be cooled to about 0 °C and then held at that temperature for at least 16 hours. Synthesis of a of Formula (VI)

[0148] The present application also provides a compound of Formula (VI):

[0149] Formula (VI) wherein wherein, R2is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0150] In some embodiments,

[0151] In some embodiments,

[0152] In some embodiments,

[0153] In some embodiments, R2is a C1-6 alkyl. In some embodiments, R2is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R2is CH3.

[0154] In some embodiments, the process for preparing the compound of Formula (VI): comprises sulfonylation of an anthranilate of Formula (VII) using a nosylating agent: wherein wherein, R2is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0155] In some embodiments,

[0156] In some embodiments,

[0157] In some embodiments,

[0158] In some embodiments, R2is a C1-6 alkyl. In some embodiments, R2is selected from methyl, ethyl, zz-propyl, z-propyl, zz-butyl and / -butyl. In some embodiments, R2is CH3.

[0159] In some embodiments, the compound of Formula (VII) is methyl anthranilate. In some embodiments, the compound of Formula (VII) is methyl anthranilate and the compound of Formula (VI) is a compound of Formula (Via) as described herein.

[0160] In some embodiments, the process for preparing the compound of Formula (Via): comprises sulfonylation of methyl anthranilate using a nosylating agent:

[0161] In some embodiments, the nosylating agent is 2-nitrobenzenesulfonyl chloride, 4- nitrobenzenesulfonyl chloride or 2, 4-nitrobenzenesulfonyl chloride. In some embodiments, the nosylating agent is 4-nitrobenzenesulfonyl chloride (also referred to as 4-nosylchloride). Any suitable amount of nosylating agent may be used. In some embodiments, the nosylating agent is provided in molar equivalents of between about 0.01 and 5, between about 0.05 and 4, or between about 0.1 and 2, or between about 0.5 to 1.5, or about 1.1 relative to the anthranilate. In some embodiments, the nosylating agent is provided in molar equivalents of about 1.1, relative to the anthranilate. The present inventors have found that increasing the amount of NsCl did not affect the reaction, and in some examples may be undesirable as it could contaminate the product.

[0162] In some embodiments, sulfonylation of the anthranilate is performed in the presence of a base. In some embodiments, the base comprises pyridine. In some embodiments, the base does not comprise dimethylaminopyridine (DMAP). In some embodiments, the base does not comprise TEA. In some embodiments, the sulfonylation of the anthranilate does not require DMAP. The base may be provided in the reaction in any amount suitable so as to effect the sulfonylation reaction. In some embodiments, the base is provided in molar equivalents, relative to the molar amount of the anthranilate, of less than about 5, 4, 3, 2, 1.5, 1.0, or 0.5. In some embodiments, the base is provided in molar equivalents, relative to the molar amount of the anthranilate, of greater than about 0.01, 0.05, 0.1, 0.3, 0.5, 1, 1.5, 2, or 3. The base may be in a range provided by any two of these upper and / or lower ranges, for example between about 0.01 and 5, between about 0.05 and 4, or between about 0.1 and 2, or between about 0.5 to 1.5. In some embodiments, the base is pyridine and provided in molar equivalents, relative to the molar amount of the anthranilate, of between 0.5 and 3, or between 1.5 and 2.5 or of about 2. In some embodiments, the base is pyridine and provided in molar equivalents, relative to the molar amount of the anthranilate, of about 2. The present inventors have found that the use of pyridine as the base means that an additional catalyst (such as DMAP) does not have to be used. In addition, the final reaction mixture is less complex helping with isolation of the compound of formula (VI) in high yields and / or purity.

[0163] In some embodiments, sulfonylation of the anthranilate is performed in the presence of a catalyst. In some embodiments, the catalyst is dimethylaminopyridine, e.g. 4- dimethylaminopyridine. In some embodiments, the concentration of catalyst in the reaction solution is greater than about 0.001, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, or 4 mol%, relative to the amount of the anthranilate. In some embodiments, the concentration of catalyst in the reaction solution is less than about 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, or 0.01 mol %, relative to the amount of the anthranilate. The catalyst may be in a concentration range provided by any two of these upper and / or lower values, for example between about 0.001 and 5, between about 0.005 and 2, or between about 0.01 and 1. In some embodiments, the catalyst is present in an amount of about 5 mol%, relative to the amount of the anthranilate. In some embodiments, the reaction does not require a separate catalyst (such as dimethylaminopyridine) with pyridine functioning as both the base and the catalyst.

[0164] As will be appreciated by the person skilled in the art, the process will be performed in a suitable solvent. In some embodiments, the sulfonylation reaction is performed in a solvent, such as a non-polar solvent, a protic solvent, or a aprotic solvent. In some embodiments, the sulfonylation reaction is performed in a polar aprotic solvent. Examples of polar aprotic solvents include, but are not limited to, halogenated hydrocarbons, ketones, nitriles, esters, carbonate esters, ethers, sulfoxides, sulfones, amides, nitroalkanes, pyrrolidines, pyridines, and combinations thereof. Examples of nitriles include, but are not limited to, acetonitrile (MeCN). Examples of esters include, but are not limited to, ethyl formate, methyl acetate (MeOAc), ethyl acetate (EtOAc), propyl acetate, isopropyl acetate (z-PAC), n-butyl acetate, isobutyl acetate, and combinations thereof. Examples of carbonate esters include, but are not limited to, dimethyl carbonate (DMC), propylene carbonate (PC), and combinations thereof. Examples of polar and non-polar ethers include, but are not limited to, m ethyl - / c / 7-butyl ether (MTBE), diethyl ether, 1,4-di oxane, 2-methoxy ethanol, 2-ethoxy ethanol, dimethoxy ethane (DME or monoglyme), 1,1-dimethoxymethane, 2,2-dimethoxypropane, 1,1 -di ethoxypropane, isopropyl ether, petroleum ether, cyclopentyl methyl ether (CPME), anisole (methoxybenzene), methyltetrahydrofuran (MeTHF), tetrahydrofuran (THF), and combinations thereof. Examples of sulfoxides include, but are not limited to, dimethylsulfoxide (DMSO). Examples of sulfones include, but are not limited to, sulfolane. Examples of amides include, but are not limited to, formamide, V, / V-di methyl acetamide, jV, V-dimethylformamide (DMF), and combinations thereof. Examples of nitroalkanes include, but are not limited to, nitromethane. Examples of pyrrolidines include, but are not limited to, V-methyl pyrrolidone (NMP). Examples of pyridines include, but are not limited to, pyridine. In one example, the sulfonylation reaction is performed in dichloromethane (DCM). In one example, the sulfonylation reaction is performed in an alkyl nitrile. In one example, the sulfonylation reaction is performed in acetonitrile (MeCN). Use of MeCN as the solvent is thought to facilitate isolation of the reaction product.

[0165] In some embodiments, the solvent may contain water. For example, the solvent may be aqueous acetonitrile.

[0166] The solvent may be present in the reaction in any amount suitable so as to effect the sulfonylation reaction. In some embodiments, the solvent is present in volume equivalents (L), relative to the molar amount of the anthranilate, of less than about 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1. In some embodiments, the solvent is present in volume equivalents (L), relative to the molar amount of the anthranilate, of more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 25. The solvent may be in a range provided by any two or more of the upper and / or lower amounts, for example between about 1 and 30, between about 2 and 20, between about 5 and 10 or between about 7 and 9. In one example, the solvent is present in volume equivalents (L), relative to the molar amount of the anthranilate, of about 8. The volume equivalent of solvent may be selected to provide an appropriate reaction rate, while ensuring the reaction mixture does not become too difficult to stir due to the formation of a precipitate. In one example, the solvent is acetonitrile and present in volume equivalents (L), relative to the molar amount of the anthranilate, of about 8.

[0167] The person skilled in the art will appreciate that it may be necessary to control the temperature of the reaction. In one example, the temperature of the reaction is maintained at or below 35 °C, at or below 30 °C. In one example, the sulfonylation reaction is maintained at about 25 °C. In some embodiments, the reaction is allowed to proceed for at least one hour, at least 2 hours, at least three hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 8 hours or at least 10 hours. The person skilled in the art will be able to monitor the reaction to determine when the reaction is sufficiently complete. The present inventors have found that unreacted methyl anthranilate is readily rejected from the isolated product in amounts as high as 10 mol%.

[0168] After completion of the reaction, the compound of formula (VI) may be isolated prior to being used in the next step. Any suitable technique known to the person skilled in the art, e.g. filtration, chromatography, crystallisation and the like, may be used. In some embodiments, the compound of formula (VI) is isolated by filtration. In some embodiments, reaction mixture is diluted with water before filtration to obtain the compound of Formula (VI). In some embodiments, the water is present in volume equivalents (L), relative to the molar amount of the anthranilate, of less than about 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1. In some embodiments, the water is present in volume equivalents (L), relative to the molar amount of the anthranilate, of more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, or 25. The water may be in a range provided by any two or more of the upper and / or lower amounts, for example between about 1 and 30, between about 2 and 20, between about 5 and 10 or between about 7 and 9. In one example, the water is present in volume equivalents (L), relative to the molar amount of the anthranilate, of about 8. In some embodiments, the water is added in about an equi-volume amount relative to the solvent.

[0169] Synthesis of a Compound of Formula (III)

[0170] The present application also provides an enantiomerically pure compound of formula

[0171] (III):

[0172] Formula (III) wherein wherein R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl; and wherein R2is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0173] In some embodiments,

[0174] In some embodiments,

[0175] In some embodiments,

[0176] In some embodiments, R1is a C1-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R1is CH3.

[0177] In some embodiments, R2is a C1-6 alkyl. In some embodiments, R2is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R2is CH3. In some embodiments, R1is CH3, and R2is CH3.

[0178] In some embodiments, the compound of formula (III) is a compound of formula (Illa):

[0179] Formula (Illa) In other words,

[0180] The present application also provides a process for preparing a compound of Formula (III). In some embodiments, the compound of Formula (III) is an enantiomerically pure compound of Formula (III). For example, in some embodiments, the process provides a compound of formula (III) having an enantiomeric purity of at least 90%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% R.

[0181] In some embodiments, the process for preparing the compound of Formula (III) comprises alkylation of a compound of formula (IV) with a compound of formula (VI) to form a compound of formula (III): wherein R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl; and wherein R2is an optionally substituted C1-6 alkyl or an optionally substituted C3-7 cycloalkyl. In some embodiments,

[0182] In some embodiments,

[0183] In some embodiments,

[0184] In some embodiments, R1is a Ci-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R1is CHa.

[0185] In some embodiments, R2is a Ci-6 alkyl. In some embodiments, R2is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R2is CH3.

[0186] In some embodiments, R1is CH3, and R2is CH3.

[0187] In some embodiments, the compound of Formula (IV) is a compound of Formula (IVa). In some embodiments, the compound of Formula (III) is a compound of Formula (Illa).

[0188] In some embodiments, the process for preparing the compound of Formula (III) comprises a Mitsunobu reaction (e.g. a Fukuyama-Mitsunobu reaction) of a compound of formula (IV) with a compound of formula (VI) to form a compound of formula (III). In the a Mitsunobu reaction, the chiral alcohol undergoes an inversion of stereochemistry. As would be appreciated by the person skilled in the art the Mitsunobu reaction converts an alcohol (such as a compound of Formula (IV) or (IVa)) using triphenylphosphine and an azodicarboxylate. In Suitable azodicarboxylates include, but are not limited to, dialkyl azodicarboxylates such as diethyl azodicarboxylate (DEAD) or diisopropyl azodicarboxylate (DIAD). In some embodiments, the Mitsunobu reaction is performed in the presence of a triphenylphosphine (TPP) and diethylazodicarboxylate (DEAD). In some embodiments, the Mitsunobu reaction is performed in the presence of triphenylphosphine (TPP) and diisopropyl azodicarboxylate (DIAD).

[0189] TPP and the azodicarboxylate may be provided in the reaction in any amount suitable so as to effect the Mitsunobu reaction. In some embodiments, TPP is provided in molar equivalents, relative to the molar amount of the compound of Formula (IV), of less than about 5, 4, 3, 2, 1.5, 1.25, or 1.0. In some embodiments, the TPP is provided in molar equivalents, relative to the molar amount of the compound of Formula (IV), of greater than about 0.5, 0.8, 1, 1.25, 1.5 or 2. TPP may be in a range provided by any two of these upper and / or lower ranges, for example between about 0.01 and 5, between about 0.05 and 4, or between about 0.1 and 2, or between about 0.5 to 1.5. In some embodiments, TPP is provided in molar equivalents, relative to the molar amount of the compound of Formula (IV), of about 1.5. In some embodiments, the azodicarboxylate (e.g. DIAD) is provided in molar equivalents, relative to the molar amount of the compound of Formula (IV), of less than about 5, 4, 3, 2, 1.5, 1.25, or 1.0. In some embodiments, the azodicarboxylate (e g. DIAD) is provided in molar equivalents, relative to the molar amount of the compound of Formula (IV), of greater than about 0.5, 0.8, 1, 1.25, 1.5 or 2. The azodicarboxylate (e.g. DIAD) may be in a range provided by any two of these upper and / or lower ranges, for example between about 0.01 and 5, between about 0.05 and 4, or between about 0.1 and 2, or between about 0.5 to 1.5. In some embodiments, the azodicarboxylate (e.g. DIAD) is provided in molar equivalents, relative to the molar amount of the compound of Formula (IV), of about 1.5.

[0190] As would be appreciated by the person skilled in the art, the alkylation reaction is performed in the presence of a solvent. Any suitable solvent may be used, such as those described herein. In some embodiments, the solvent is a aprotic solvent or non-polar solvent as described herein. In some embodiments, the aprotic solvent or non-polar solvent is selected from diethyl ether, dichloromethane (DCM), tetrahydrofuran (THF), methyltetrahydrofuran (MeTHF), acetonitrile (MeCN), / V-methylpyrrolidone (NMP), pyridine, toluene, hexanes, n- heptane, ethyl acetate (EtOAc), methylisopropyl ketone (MIPK), 7V, / V-dimethylformamide (DMF), dimethylsulfoxide (DMSO), or any combinations thereof. In some embodiments, the solvent is dichloromethane (DCM). In some embodiments, the amount of solvent is present in volume equivalents (L), relative to the molar amount of the compound of Formula (IV), of between about 1 and 30, between about 2 and 20, or between about 5 and 10, or between about 5 and 7, or about 5.

[0191] In some embodiments, the concentration of the compound of Formula (IV) in the reaction solution (mol / L) is greater than about 0.001, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, or 4. In some embodiments, the concentration of the compound of Formula (IV) in the reaction solution (mol / L) is less than about 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, or 0.01. The compound of Formula (IV) may be in a concentration range provided by any two of these upper and / or lower values, for example between about 0.001 and 5, between about 0.005 and 2, or between about 0.01 and 1.

[0192] In some embodiments, the concentration of the compound of Formula (VI) in the reaction solution (mol / L) is greater than about 0.001, 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, or 4. In some embodiments, the concentration of the compound of Formula (VI) in the reaction solution (mol / L) is less than about 5, 4, 3, 2, 1, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, 0.05, or 0.01. The compound of Formula (VI) may be in a concentration range provided by any two of these upper and / or lower values, for example between about 0.001 and 5, between about 0.005 and 2, or between about 0.01 and 1.

[0193] In some embodiments, the compound of formula (VI) is provided in molar equivalents of between about 0.5 and 5, between about 0.8 and 3, between about 1 and 2 or between about 1.0 and 1.5, or about 1.1, relative to the compound of formula (IV).

[0194] In some embodiments, the order of addition of the reagents of the Mitsunobu reaction (e g. Fukuyama-Mitsunobu reaction) may be important. In some embodiments, the compound of formula (IV), the compound of formula (VI), and triphenylphosphine are dissolved in a suitable solvent, cooled to less than 0 °C before addition of the azodicarboxylate (e.g. DIAD). In some embodiments, the reaction mixture is cooled to about -5 °C prior to the addition of the azodi carb oxy late (e.g. DIAD). In some embodiments, the reaction mixture is maintained at 0 °C or less during the addition of DIAD. In some embodiments, the reaction mixture is maintained at 5 °C or less during the addition of DIAD. In some embodiments, the reaction mixture is maintained at 8 °C or less during the addition of DIAD. In some embodiments, the reaction mixture is maintained for at least 30, 40, 50, 0, 70, 77 or 80 minutes. In some embodiments, the reaction mixture is then raised to between about 15 °C and 30 °C, or about 25 °C over a period of time (e g. at least 1, 2, 3, 4, or 5 hours). In some embodiments, the reaction mixture is then stirred at room temperature for several hours (e.g. at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 hours). In some embodiments, following addition of azodi carb oxy late (e g. DAID) the reaction is heated to between about 15 °C and 30 °C, about 20 °C and 25 °C, or about 25 °C. In one example, the Mitsunobu reaction is heated to about 25 °C for at least 10 hr, at least 12 hr, at least 14 hr, at least 16 hr, at least 20 hr or at least 24 hrs. In one example, the Mitsunobu reaction is performed in DCM and heated to between about 25 °C for a period of 16 hr.

[0195] Once the reaction is complete, the compound of formula (III) may be isolated. The present inventors have found that solvent exchange into a first solvent (such as THF) precipitates the racemate compound of Formula (III). This precipitate may be filtered and discarded. Including this first solvent exchange step is thought to improve the enantiomeric purity of the compound of Formula (III) as the solvent exchange and incubation steps selectively precipitate the racemate compound of Formula (III). In some embodiments, after solvent exchange the reaction mixture is incubated (e.g. at 20°C), optionally with stirring, for least 30 min, at least 60 min, at least 90 min, at least 2 hr prior to filtration. In some embodiments, after the first incubation, the reaction mixture may be further cooled (e.g. to 5°C) and incubated for at least a further 1, 2, 3, 4, or 5 hours, prior to filtration. In some embodiments, after the second incubation (e.g. at 5°C), the reaction mixture may be further incubated (e g. 10°C) for at least a further 2, 4, 6, 8, 10, 12, 14, 16, 18 or 20 hours, prior to filtration. It is thought the additional incubation times improves removal of the racemate. In some embodiments, a filtration aid (such as Seitz K200 clear-filtration aid) is added prior to filtration to remove rac- Formula (III). The present inventors have also found that solvent exchange of the filtrate into a second solvent (e.g. an alcohol such as ethanol) leads to precipitation of the desired enantiomer, which may then be isolated by filtration. Accordingly, in some embodiments, isolation of the compound of formula (III) comprises: a first solvent exchange into a first solvent, filtration to obtain a filtrate, a second solvent exchange into an alcohol to precipitate the compound of formula (III); and isolation of the precipitate. In some embodiments, the first solvent is a THF. In some embodiments, the alcohol is selected from methanol, ethanol and isopropyl alcohol, or any combinations thereof. In some embodiments, the alcohol is ethanol. In some embodiments, following solvent exchange into ethanol, the solution is cooled gradually (e.g. from +70°C to +20°C over 300 min; 10°C / h). Once crystallisation is observed, the mixture may be incubated for a further 2, 4, 6, 8, 10, 12 or 13 or more hours before filtration. Finally, the present inventors also found that the optical purity of the final product could be improved by longer holding time during racemate precipitation, and the use of polish filtration aid to assist racemate removal.

[0196] In some embodiments, the process provides a compound of formula (IV) having an enantiomeric purity of at least 90%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%. Synthesis of a Compound of Formula (I

[0197] The present application also provides an enantiomerically pure compound of Formula (I): wherein R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0198] In some embodiments, R1is a C1-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R1is CH3. In some embodiments, the compound of formula (I) has an enantiomeric purity of at least 90%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%.

[0199] The present application also provides a process for preparing a compound of Formula (I). In some embodiments, the compound of Formula (I) is an enantiomerically pure compound of Formula (I). For example, in some embodiments, the process provides a compound of formula (I) having an enantiomeric purity of at least 90%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% R.

[0200] In some embodiments, the process comprises converting the compound of Formula (III) to a compound of Formula (I). wherein R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl; and wherein R2is an optionally substituted C1-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0201] In some embodiments,

[0202] In some embodiments,

[0203] In some embodiments,

[0204] In some embodiments, R1is a C1-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R1is CH3.

[0205] In some embodiments, R2is a C1-6 alkyl. In some embodiments, R2is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R2is CH3. In some embodiments, R1is CI I3, and R2is CHa.

[0206] In some embodiments, converting the compound of Formula (III) to a compound of Formula (I) occurs via an intermediate compound of Formula (II). In some embodiments, the compound of Formula (II) is isolated prior to the consequent reaction. In some embodiments, the compound of Formula (II) is not isolated prior to the consequent reaction. In some embodiments, the compound of Formula (III) is a compound of Formula (Illa), the compound of Formula (II) is a compound of Formula (Ila), and the compound of Formula (I) is a compound of Formula (la), as defined herein. In some embodiments, the present application also provides a process for preparing an enantiomerically pure compound of Formula (I):

[0207] Formula (I) the method comprising:

[0208] (a) deprotecting a compound of Formula (III) to form a compound of Formula (II); and

[0209] (b) ester hydrolysis of the compound of Formula (II) to form the compound of Formula

[0210] (I)

[0211] Formula (II) Formula (I) wherein R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl; and wherein R2is an optionally substituted C1-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0212] In some embodiments,

[0213] In some embodiments,

[0214] In some embodiments,

[0215] In some embodiments, R1is C1-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R1is CHa.

[0216] In some embodiments, R2is C1-6 alkyl. In some embodiments, R2is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R2is CHa.

[0217] In some embodiments, R1is CH3, and R2is CH3.

[0218] As would be appreciated by the person skilled in the art, deprotecting the compound of Formula (III) comprises removal of the nosyl (nitrobenzenel sulfonyl) group (i.e. denosylation). The nosyl group may be deprotected using any suitable technique known to the person skilled in the art. In some embodiments, the nosyl group is deprotected by aromatic nucleophilic substitution with a thiol. In some embodiments, the nosyl group is removed using a thiol reagent in the presence of a base. In some embodiments, the thiol reagent is selected from thiophenol or thioglycolic acid. In some embodiments, the thiol reagent is thioglycolic acid. An advantage of thioglycolic acid is the fact that the cleavage side product, nitrophenylthioacetic acid, can be washed away from the product with aqueous Nal ICOs and there is lower impurity formation in the reaction. Any suitable base may be used. In some embodiments, the base comprises a piperidine, a guanidine or an amidine base. In some embodiments, the base comprises 1,8- Diazabicyclo[5.4.0]undec-7-ene (DBU) or tetramethylguanidine (TMG) or combinations thereof. In some embodiments, the base comprises TMG. In some embodiments, the selected base does not promote nucleophilic substitution or elimination of the starting material which may help reduce impurities.

[0219] The base and / or the thiol reagent may be provided in the reaction in any amount suitable so as to effect deprotection. In some embodiments, the thiol reagent is provided in molar equivalents, relative to the molar amount of the compound of Formula (III), of less than about 5, 4, 3, 2, 1.5, 1.2, 1.1 or 1.0. In some embodiments, the thiol reagent is provided in molar equivalents, relative to the molar amount of the compound of Formula (III), of greater than about 0.01, 0.05, 0.1, 0.3, 0.5, 1, 1.3, 1.5, 2, or 3. The thiol reagent may be in a range provided by any two of these upper and / or lower ranges, for example between about 0.01 and 5, between about 0.05 and 4, or between about 0.1 and 2, or between about 0.5 to 1.5. In some embodiments, the thiol reagent is provided in molar equivalents, relative to the molar amount of the compound of Formula (II), of 1.5. Under the HSCH2COOH / TMG conditions, thioglycolic acid loading was about 1.5 mol equiv. relative to the molar amount of the compound of Formula (III).

[0220] In some embodiments, the base is provided in molar equivalents, relative to the molar amount of the compound of Formula (III), of less than about 10, 8, 5, 4, 3, 2.5, 2, 1.5, or 1.0. In some embodiments, the base is provided in molar equivalents, relative to the molar amount of the compound of Formula (III), of greater than about 0.01, 0.05, 0.1, 0.3, 0.5, 1, 1.3, 1.5, 2, 3, 4 or 5. The base may be in a range provided by any two of these upper and / or lower ranges, for example between about 0.5 and 8, between about 2 and 6, or between about 2.5 and 5. In some embodiments, the base is provided in molar equivalents, relative to the molar amount of the compound of Formula (II), of about 3. Under the HSCH2COOH / TMG conditions, about 3.0 mol equiv of base was preferred, relative to the molar amount of the compound of Formula (III).

[0221] In some embodiments, the molar ratio of thiol :base is between 5: 1 and 1:5, or between 1: 1 and 1:3, or between 1: 1.5 and 1 :2.5. In some embodiments, the molar ratio of HSCH2COOH / TMGis between 5:1 and 1:5, or between 1 : 1 and 1 :3, or between 1:1.5 and 1:2.5, or between 1 :1.8: 1:2.2, or about 1 :2. In some embodiments, the molar ratio of HSCH2COOH / TMG ratio is about 1:2, consistent with double deprotonation of thioglycolic acid being crucial for ensuring its reactivity.

[0222] In some embodiments, the deprotection reaction is performed in the presence of a solvent. In some embodiments, the solvent is a polar aprotic solvent or a non-polar as defined herein. In some embodiments, the aprotic solvent or non-polar solvent is selected from di chloromethane (DCM), tetrahydrofuran (THF), methyltetrahydrofuran (MeTHF), acetonitrile (MeCN), / V-m ethyl pyrrolidone (NMP), pyridine, toluene, hexanes, n-heptane, ethyl acetate (EtOAc), methylisopropyl ketone (MIPK), A( / V-di methyl form am ide (DMF), dimethylsulfoxide (DMSO), acetone, or any combinations thereof. In some embodiments, the solvent is a polar aprotic solvent, for example, acetone, acetonitrile, dichloromethane, DMSO or tetrahydrofuran or combinations thereof. In some embodiments, the solvent is acetonitrile or acetone or combinations thereof. In some embodiments, the solvent is acetonitrile. In some embodiments, the amount of solvent is present in volume equivalents (L), relative to the molar amount of the compound of Formula (III), of between about 1 and 30, between about 2 and 20, or between about 3 and 10, or between about 3 and 8, or about 3 and 5, or about 5.

[0223] In some embodiments, the deprotection reaction is incubated for at least 2, 4, 6, 8, 10, 12, 14, 16, 17, 18, 19 or 20 hours at a suitable temperature (e.g. 25 °C). In some embodiments, the deprotection reaction is incubated for at least 17 hours at a suitable temperature (e g. 25 °C). If the reaction is still incomplete after the first incubation, further HSCH2COOH / TMG may be added and the reaction mixture incubated for at least a further 2, 4, 6, 8, 10, 12, 14, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 or 26 hours.

[0224] Once complete, the reaction may be diluted with a second solvent (e g. MTBE (methyl tert-butyl ether) or DCM) and washed with water and / or sodium bicarbonate solution to remove water-soluble nosyl deprotection byproducts. In some embodiments, the second solvent is MTBE. The present inventors have found that MTBE offers several advantages, including better layer separation during extraction and an operation sequence that prevents oiling out of the reaction mixture as the second solvent is added. In some embodiments, the second solvent is DCM. In some embodiments, the amount of second solvent is present in volume equivalents (L), relative to the molar amount of the compound of Formula (III), of between about 1 and 30, between about 2 and 20, or between about 3 and 10. In some embodiments, the diluted reaction mixture may be washed with water and / or sodium bicarbonate solution to remove water-soluble nosyl deprotection byproducts. In some embodiments, the product is obtained as a solution in solvent (e.g. DCM or MTBE) and is transferred to the next step directly. In some embodiments, the product is obtained as a solution in solvent and used in the next step immediately. In some embodiments, the product is isolated before being used in the next step using techniques known to the person skilled in the art. In embodiments where the product is used in the next step without isolation, the solvent is exchanged to a suitable solvent such as an alcohol, for example methanol, ethanol or isopropyl alcohol, or any combinations thereof. In some embodiments, the alcohol is methanol. Any suitable technique for solvent exchange may be used. In some embodiments, the product is concentration by distillation before being diluted in the alcohol. The distillation process is repeated at least one, two or three times to obtain the compound of Formula (II) in solution with the alcohol.

[0225] Following solvent exchange, the ester is hydrolysed using a base to form a compound of Formula (I). Any suitable base may be used. In some embodiments, the base is sodium hydroxide. The person skilled in the art will appreciate that it may be necessary to apply heat to facilitate the hydrolysis reaction. The amount of heat required may depend upon the solvent in which the reaction is performed in, as discussed above. In some embodiments, the hydrolysis reaction is heated to between about 30 °C and 100 °C, about 40 °C and 90 °C, about 60 °C and 80 °C, or about 70 °C and 80 °C. In one example, the hydrolysis reaction is performed in methanol - aqueous sodium hydroxide mixture and heated to between about 75 °C and 80 °C. The duration of heating is sufficient for the reaction to go to completion. In some embodiments, the reaction is heated for at least 0.5 hr, at least 1 hr, or at least 1.5 hr. In some embodiments, the reaction is heated for at least Ihr. Following completion of the hydrolysis reaction, the base may be neutralised with an acid. In some embodiments, an acid (e.g. acetic acid) is added during the workup step to form the compound of Formula (I). In some embodiments, the acid is added such that the final pH of the solution is 7.5 or lower, 7 or lower, 6 or lower, 5 or lower or between 4 and 5. In some embodiments, the acetic acid is added until the pH of the solution is between about 4 and 5.

[0226] In some embodiments, the compound of Formula (I) may be isolated following acidification of the reaction. The compound of Formula (I) may be isolated according to any suitable means as would be understood by the person skilled in the art. For example, isolation of the compound of Formula (I) may be achieved by filtration, chromatography (e.g. reversephase or normal phase column chromatography), distillation, extraction, or recrystallization. In some embodiments, the alcohol (e.g. methanol) may be removed by distillation. In some embodiments, distillation to remove the alcohol may be followed by a series of extractions with a solvent such as DCM. In some embodiments, the final product may be precipitated following solvent exchange into a solvent with lower solubility for the compound of Formula (I). In some embodiments, the final product may be precipitated following solvent exchange into acetonitrile. In some embodiments, seeding may be used during solvent exchange to control precipitation of the compound of Formula (I). The compound of Formula (I) may then be isolated by filtration. In some embodiments, the solution is cooled (e g. from +80 °C to +20 °C) over a suitable period of time (e.g. at least 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 h). In some embodiments, the solution is cooled to +20 °C at a rate of 10°C / h. In some embodiments, the cooled mixture is incubated for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15h.

[0227] In some embodiments, the step of isolating the compound of Formula (I) is achieved by recrystallization. In recrystallising the compound of Formula (I), the person skilled in the art would appreciate that a suitable solvent includes that in which the compound of Formula (I) has a reduced solubility. In some embodiments, the step of isolating the compound of Formula (I) is achieved by recrystallization wherein the solvent is selected from dichloromethane, n- heptane, ethanol, isopropyl alcohol, toluene, ethyl - / -butyl ether, acetone, methylethyl ketone, isopropyl acetate, and combinations thereof. In one example, the step of isolating the compound of Formula (I) is achieved by recrystallization wherein the solvent is a combination of isopropyl alcohol and dichloromethane. In one example, the step of isolating the compound of Formula (I) is achieved by recrystallization wherein the solvent is a combination of ethanol and di chloromethane. In one example, the step of isolating the compound of Formula (I) is achieved by recrystallization wherein the solvent is a combination of methanol and dichloromethane. In one example, the step of isolating the compound of Formula (I) is achieved by recrystallization wherein the solvent is isopropanol. In one example, recrystallization comprises filtration with a filtration aid, such as Seitz K200 filter aid. Without wishing to be bound by theory, the present inventors have found that the racemate is less soluble than the pure enantiomer. Accordingly, filtration is thought to help remove impurities such as the racemate and inorganic impurities. In some embodiments, the crude compound of Formula (I) is dissolved in di chloromethane, filtered to remove any precipitate (optionally including any racemate), solvent exchanged into an alcohol (e.g. isopropanol or ethanol), stirred, optionally seeded and then filtered to obtain the compound of Formula (I). In some embodiments, the crude compound of Formula (I) is dissolved in dichloromethane, filtered to remove any precipitate (optionally including any racemate), solvent exchanged into ethanol, stirred, optionally seeded and then filtered to obtain the compound of Formula (I). In some embodiments, the crude compound of Formula (I) is dissolved in dichloromethane, filtered to remove any precipitate (optionally including any racemate), solvent exchanged into aqueous ethanol, stirred, optionally seeded and then filtered to obtain the compound of Formula (I). In some embodiments, the amount of di chloromethane is present in volume equivalents (L), relative to the molar amount of the compound of Formula (I), of between about 5 and 20, between about 8 and 15, or between about 9 and 12, or about 10. In some embodiments, the amount of alcohol is present in volume equivalents (L), relative to the molar amount of the compound of Formula (I), of between about 5 and 20, between about 8 and 15, or between about 9 and 12, or about 10. In some embodiments, the solvent exchanged mixture is seeded at a temperature of at least 50 °C, at least 55 °C, at least 60 °C, at least 65 °C or at least 70 °C. In some embodiments, water is added after solvent exchange. In some embodiments, water is added after seeding. In some embodiments, the crystallisation mixture is cooled to less than 5 °C, e.g. 0 °C and held at that temperature to promote crystal formation.

[0228] In some embodiments, the present application also provides a process for preparing an enantiomerically pure compound of Formula (I):

[0229] Formula (I) which uses an enantiomerically pure compound of Formula (III) as defined herein as an intermediate. R1is as defined herein.

[0230] In some embodiments, the present application also provides a process for preparing an enantiomerically pure compound of Formula (I): Formula (I) which uses an enantiomerically pure compound of Formula (IV) as defined herein as an intermediate. R1is as defined herein.

[0231] In some embodiments, the present application also provides a process for preparing an enantiomerically pure compound of Formula (I):

[0232] Formula (I) the process comprising:

[0233] (a) converting a compound of formula (V) to a compound of formula (IV)

[0234] Formula (V) Formula (IV)

[0235] (b) alkylation of a compound of formula (IV) with a compound of formula (VI) to form a compound of formula (III):

[0236]

[0237] (c) deprotecting a compound of Formula (III) to form a compound of Formula (II); and (d) ester hydrolysis of the compound of Formula (II) to form the compound of

[0238] Formula (I)

[0239] Formula (IT) Formula (I) wherein wherein R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl; and wherein R2is an optionally substituted C1-6 alkyl or an optionally substituted C3-7 cycloalkyl.

[0240] In some embodiments,

[0241] In some embodiments,

[0242] In some embodiments,

[0243] In some embodiments, R1is C1-6 alkyl. In some embodiments, R1is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R1is CH3.

[0244] In some embodiments, R2is C1-6 alkyl. In some embodiments, R2is selected from methyl, ethyl, n-propyl, i-propyl, n-butyl and t-butyl. In some embodiments, R2is CHa.

[0245] In some embodiments, R1is CH3, and R2is CH3.

[0246] In some embodiments, the process provides a compound of formula (I) having an enantiomeric purity of at least 90%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% R.

[0247] In some embodiments, the compound of Formula (I) is a compound of Formula (la):

[0248] Formula (la)

[0249] In some embodiments, the compound of Formula (II) is a compound of Formula (Ila):

[0250] In some embodiments, the compound of Formula (III) is a compound of Formula (Illa): wherein Ns is as defined herein. Compounds

[0251] In some embodiments, there is provided an enantiomerically pure compound of Formula (IV): wherein R1is as described herein. In some embodiments, the compound of Formula (IV) is prepared by any one or more of the processes as described herein.

[0252] In some embodiments, there is provided an enantiomerically pure compound of Formula

[0253] (IVa):

[0254] In some embodiments, the compound of Formula (IVa) is prepared by any one or more of the processes as described herein.

[0255] In some embodiments, there is provided an enantiomerically pure compound of Formula

[0256] (Ill):

[0257] Formula (III); wherein Ns, R1and R2are as described herein. In some embodiments, the compound of Formula (III) is prepared by any one or more of the processes as described herein.

[0258] In some embodiments, there is provided an enantiomerically pure compound of Formula (Illa):

[0259] Formula (Illa). In some embodiments, the compound of Formula (Illa) is prepared by any one or more of the processes as described herein.

[0260] In some embodiments, there is provided an enantiomerically pure compound of Formula (II):

[0261] Formula (II); wherein R1and R2are as described herein. In some embodiments, the compound of Formula (II) is prepared by any one or more of the processes as described herein.

[0262] In some embodiments, there is provided an enantiomerically pure compound of Formula (Ila):

[0263] Formula (Ila). In some embodiments, the compound of Formula (Ila) is prepared by any one or more of the processes as described herein.

[0264] In some embodiments, there is provided an enantiomerically pure compound of Formula (I):

[0265] Formula (I); wherein R1and R2are as described herein. In some embodiments, the compound of Formula (I) is prepared by any one or more of the processes as described herein.

[0266] In some embodiments, there is provided an enantiomerically pure compound of Formula (la):

[0267] Formula (la).

[0268] In some embodiments, the compound of Formula (la) is prepared by any one or more of the processes as described herein.

[0269] The invention further relates to an enantiomerically pure compound of Formula (I) as described herein, or pharmaceutically acceptable salts thereof, being in a solid state which can be amorphous, at least partly crystalline or substantially crystalline. The crystalline form may be more stable, easier to handle and store, and easier to purify and easier to synthesise in a reproducible manner. Example solid state forms are described in herein. In some embodiments, there is provided a crystalline form. In some embodiments, there is provided a crystalline form (Form A) having an XRPD pattern having peaks at one or more of the two theta values shown in Figure 1 (20 degrees±0.2). In some embodiments, there is provided a crystalline form having an XRPD pattern having peaks at two or more, three or more, four or more, five or more or all of the two theta values shown in Figure 1 (20 degrees±0.2). In some embodiments, there is provided a crystalline form having an XRPD pattern having peaks all of the two theta values shown in Figure 1 (20 degrees±0.2). In some embodiments, there is provided a crystalline form having an XRPD pattern that is substantially the same as shown in Figure 1. It will be understood that the 2-theta values of the X-ray powder diffraction patterns for crystalline form (A) may vary slightly from one instrument to another and also depending on variations in sample preparation and batch to batch variation, and so the values quoted are not to be construed as absolute. It will also be understood that the relative intensities of peaks may vary depending on orientation effects so that the intensities shown in the XRPD trace included herein are illustrative and not intended to be used for absolute comparison. Accordingly, it is to be understood that the phrase “substantially the same XRPD pattern as shown in Figure 1” means that for comparison purposes, at least 90% of the peaks shown in Figure 1 are present. It is to be understood that the relative peak positions may vary ±0.2 degrees from the peak positions shown in Figure 1. It is to be further understood that for comparison purposes some variability in peak intensities from those shown in Figure 1 is allowed.

[0270] In some embodiments, the crystalline form has substantially the same crystal structure shown in Figure 2.

[0271] In some embodiments, the compound of Formula (I) (e.g. the enantiomerically pure compound of Formula (I)) has substantially the same Infrared Spectrum (IR spectrum) as shown in Figure 3.

[0272] In some embodiments, the compound of Formula (I) (e.g. the enantiomerically pure compound of Formula (I)) has substantially the same Differential scanning calorimetry (DSC) thermogram as shown in Figure 4. In some embodiments, the DSC thermogram comprises one endothermic event with an on set temperature at 255.0 °C.

[0273] Compositions

[0274] Whilst a compound of Formula (I) or salt thereof may in some embodiments be administered alone, it is more typically administered as part of a pharmaceutical composition or formulation. Thus, the present disclosure also provides a pharmaceutical composition comprising a compound of Formula (I) or salt thereof and a pharmaceutically acceptable excipient. The pharmaceutical composition comprises one or more pharmaceutically acceptable diluents, carriers, or excipients (collectively referred to herein as “excipient” materials).

[0275] The present disclosure also provides pharmaceutical formulations or compositions, both for veterinary and for human medical use, which comprise compounds of Formula (I) of the present disclosure or a pharmaceutically acceptable salt thereof, with one or more pharmaceutically acceptable carriers, and optionally any other therapeutic ingredients, stabilizers, or the like. The carrier(s) must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the formulation and not unduly deleterious to the recipient thereof.

[0276] The pharmaceutical formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy. All methods include the step of bringing a compound of Formula (I) or salt thereof into association with the excipient that constitutes one or more necessary ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired formulation.

[0277] In some embodiments, the compound of Formula (I) is a compound of Formula (la).

[0278] In some embodiments, there is provided a composition comprising the enantiomerically pure compound of Formula (la) and one or more excipients according to any embodiments or examples thereof as described herein:

[0279] Formula (la); wherein any impurities, if present, are in an amount (weight % of the amount of the compound of Formula (la)) of less than about 5, 4, 3, 2, 1, 0.5, 0.1, 0.05, 0.01, 0.005, 0.001, 0.0005 or 0.0001. The composition may be substantially free of any impurities. The impurities may be selected from any one or more of the by-products or reagents used in the processes as described herein. The composition may be a pharmaceutical composition comprising one or more pharmaceutically acceptable excipients according to any embodiments or examples thereof as described herein. In some embodiments, composition comprises a compound of formula (la) having an enantiomer ratio of at least 90%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99%. In some embodiments, the enantiomerically pure compound of Formula (la) may be in a neutral form. The neutral form may be more stable, easier to handle and store, easier to purify and easier to synthesise in a reproducible manner.

[0280] It will be appreciated by persons skilled in the art that numerous variations and / or modifications may be made to the above-described embodiments, without departing from the broad general scope of the present disclosure. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive.

[0281] Any discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is not to be taken as an admission that any or all of these matters form part of the prior art base or were common general knowledge in the field relevant to the present disclosure as it existed before the priority date of each of the appended claims.

[0282] Examples

[0283] The invention disclosed herein will now be further described by reference to the following non-limiting examples.

[0284] Example 1: Synthesis of the compound of Formula (la)

[0285] The compound of Formula (la) was prepared on multi-gram scale via the intermediate compound of Formula (IVa), providing quantities (e.g. 92 g) of the compound of Formula (la) suitable for clinical development. Individual steps of the synthetic route and synthesis of the intermediate compounds disclosed herein were as follows.

[0286] Notation used

[0287] Chiral purity (also referred to as enantiomeric purity) of products is expressed as e.r. or relative area of the desired enantiomer by chiral HPLC analysis (%-area); these terms are used interchangeably. HPLC chiral purity of 50 %-area indicates a racemic mixture. The concept of ee (enantiomeric excess) is not used in the following. Step (a): (5)-9-(l-Hydroxyethyl)-7-methyl-2-morpholino-4H-pyrido[l,2-a]pyrimidin-4- one

[0288] A compound of Formula (IVa) was prepared from a compound of Formula (Va) by asymmetric transfer hydrogenation in the presence of homogeneous ruthenium catalyst RuCl[(S,<S)-MsDpen](p-cymene) and a mixture of tri ethylamine (TEA) and formic acid (FA) in DCM solution under mild reflux. Formic acid was added portionwise, as this ensured faster reaction and limited side product formation. Once the reaction was finished, the product was isolated by cooling of the reaction mixture and subsequent filtration. Advantageously, the process could be performed on a 359-gram scale, furnishing the product in 55% yield and e.r. 97.2% (S).

[0289] The compound of Formula (Va) (9-acetyl-7-methyl-2-morpholin-4-yl-4 / / -pyrido[l,2- «]pyrimidin-4-one; also referred to herein as Ketone-1) may be prepared, for example, as described in W02009 / 093972 and / or W02004 / 016607 or purchased from a commercial supplier.

[0290] In a representative procedure, to a 5-L reactor, DCM (3.6 L, 10 rel. volumes) was charged, followed by the compound of Formula (Va) (359 g, 1.25 mol, 1.00 equiv.), and triethylamine (EtiN; 126 g / 174 mL, 1.25 mol, 1.00 equiv ). RuCl[(S,5)-MsDpen]( ?-cymene) catalyst (3.5 g, 0.00625 mol, 0.5 mol%, Strem Ltd., currently Ascensus Specialties) was then charged, and the reactor was evacuated and refilled with nitrogen three times. The reactor contents were then brought to gentle reflux. A solution of formic acid (69.0 g I 56.6 mL, 1.50 mol, 1.20 equiv.) in DCM (228 mL, 0.63 rel. volumes) was prepared, and then charged to the reaction mixture in six equal portions each spaced 30 minutes. Upon the addition of a fourth portion, product slurry started to form. After the addition was finished, the reaction mixture was held at reflux for additional 90 minutes. In-process control (IPC) confirmed full starting material conversion (0.2 %-area compound of Formula (V), HPLC, 265 nm). The reaction mixture was cooled to 0 °C, and then held at that temperature for 1 hour. The product slurry was filtered and washed with DCM (0.36 L, 1 rel. volume), DCM-MTBE 1 : 1 v / v (0.36 L, 1 rel. volume), and MTBE (360 mL, 1 rel. volume), in that order. The product was dried on the filter for 16 hours.

[0291] Yield: 199 g (55%), off-white solid.

[0292] Loss on drying (LOD): 0.5 %-w / w.

[0293] Purity (HPLC, 265 nm): > 99 %-area.

[0294] Enantiomeric purity (HPLC): 97.2 %-area (S).

[0295] Step (b): Methyl 2-((4-nitrophenyl)sulfonamido)benzoate

[0296] Methyl anthranilate 4-Nosylchloride Formula (Via)

[0297] The compound of Formula (Via) was prepared by sulfonation of methyl anthranilate with 4-nosyl chloride in acetonitrile solution, employing pyridine as base and catalyst. Once the reaction was complete, the reaction mixture was diluted with water, and the product was isolated by filtration.

[0298] In a representative procedure, to a 5-L reactor, MeCN (0.90 L, 6 rel. volumes) was charged, followed by 4-nosylchloride (244 g, 1.10 mol, 1.10 equiv.) and pyridine (158 g / 161 mL, 2.00 mol, 2.00 equiv.). A solution of methyl anthranilate (151 g / 129 mL, 1.00 mol, 1.00 equiv.) in MeCN (0.30 L, 2 rel. volumes) was prepared and charged to the reactor over 20 minutes, maintaining the internal temperature below 35 °C. The mixture was stirred at 25 °C for additional 4 hours. In-process control (IPC) confirmed full starting material conversion (0.9 %-area methyl anthranilate, HPLC, 265 nm). Water (1.2 L, 8 rel. volumes) was added to the reaction mixture, and the reactor contents were stirred for an additional hour. The mixture was then filtered, and the filter contents were washed twice with MeCN-water 1 : 1-v / v (2x0.30 L, 2x2 rel. volumes), then dried under suction for 16 hours.

[0299] Yield: 319 g (95%), light-yellow solid.

[0300] LOD (loss on drying): 0.5 %-w / w.

[0301] Purity (HPLC, 265 nm): > 99 %-area. Step (c): Methyl ( / ?)-2-((M-(l-(7-methyl-2-morpholino-4-oxo-4H-pyrido|l,2-u|pyrimidin-

[0302] 9-yl)ethyl)-4-nitrophenyl)sulfonamido)benzoate

[0303] The compound of Formula (Illa) was prepared via Mitsunobu reaction between the compound of Formula (IVa) and the compound of Formula (Via), mediated by triphenylphosphine (TPP) and diisopropyl azodi carb oxy late (DIAD) in DCM solvent. Once the reaction was complete, the solvent was exchanged to THF to precipitate a minor first crop of racemic version of the compound of Formula (III) (rac-compound of Formula (Illa)), which was filtered and discarded. Subsequent solvent exchange to ethanol led to precipitation of the compound of Formula (Illa) (i.e., the desired (R) enantiomer) main crop, which was isolated by filtration.

[0304] In a representative procedure, to a 5 L reactor, DCM (1.0 L. 5 rel. volumes) was charged, followed by the compound of Formula (IV) (199 g, 0.688 mol, 1.00 equiv), the compound of Formula (VI) (254 g, 0.757 mol, 1.10 equiv.), and TPP (270 g, 1.03 mol, 1.50 equiv.). The reactor contents were cooled to -5 °C, and DIAD (208 g / 204 mL, 1.03 mol, 1.50 equiv.) was charged over 67 minutes, without letting the internal temperature exceed 0 °C. The reaction mixture was held at 0 °C for additional 1 hour, warmed to 25 °C over 3 hours, then stirred at 25 °C for 16 hours. IPC confirmed full starting material conversion (the compound of Formula (IV) not detected, HPLC, 265 nm).

[0305] The reaction mixture was diluted with THF (2.0 L, 10 rel. volumes), then distilled under reduced pressure to a total volume of 2.6 L (initial volume 4.0 L). The formed thin slurry of rac-compound of Formula (III) was cooled to 25 °C and filtered on a glass frit filter equipped with Seitz K200 filter aid; the solids were rinsed with additional THF (0.20 L, 1 rel. volume) and discarded. The reactor was cleaned, and the filtrate was transferred back to the reactor. EtOH (1.6 L, 8 rel. volumes) was added, and the solvents were distilled off under atmospheric pressure to a total volume of 1.8 L; distillation was repeated with another portion of EtOH (1.6 L, 8 rel. volumes) to the end volume of 2.8 L. The mixture was cooled to 25 °C and held at that temperature for 18 hours. The slurry was filtered; the solids were washed with EtOH (2x0.40 L, 2x2 rel. volumes) and dried under air suction for 26 hours.

[0306] Yield: 320 g (77%), yellow solid.

[0307] LOD: 0.6 %-w / w.

[0308] HPLC purity: 99.6 %-area.

[0309] Enantiomeric purity (HPLC): 98.7 %-area (R) (determined in the next synthetic step due to insolubility of the compound of Formula (Illa) in conventional chiral HPLC solvents).

[0310] Step (d): Methyl ( / ?)-2-((l-(7-methyl-2-morpholino-4-oxo-4 / / -pyrido|l,2-«|pyrimidin-9- yl)ethyl)amino)benzoate

[0311] The compound of Formula (Ila) was prepared by removal of the nosyl protecting group from the compound of Formula (Illa) using a mixture of thioglycolic acid and tetramethylguanidine (TMG) in acetonitrile, and was carried forward to the next step of the synthesis without being isolated.

[0312] In a representative procedure, to a 5-L reactor, MeCN (1.4 L, 5 rel. volumes) was charged, followed by the compound of Formula (Illa) (284 g, 0.467 mol, 1.00 equiv.) and thioglycolic acid (64.4 g / 48.9 mL, 0.701 mol, 1.50 equiv.). To the resulting suspension, TMG (161 g / 176 mL, 1.40 mol, 3.00 equiv.) was added slowly over 23 minutes, maintaining internal temperature below 35 °C. After the addition was finished, the mixture was stirred at 25 °C for 3 hours. IPC1 showed incomplete starting material conversion (3.9 %-area compound of Formula (Illa)), and stirring was continued for another 20 hours, at which point IPC2 confirmed full conversion (0.4 %-area compound of Formula (Illa)). DCM (2.8 L, 10 rel. volumes) was charged to the reactor. In a separate container, a solution of NaHCOi (100 g, 35%-w / w of the compound of Formul (Illa)) in water (2.8 L, 10 rel. volumes) was prepared. The reactor contents were then washed with the prepared NaHCOi solution (2x 1.4 L, 2x5 rel. volumes), followed by water (1.4 L, 5 rel. volumes). Aqueous washings were discarded. The obtained solution of the compound of Formula (II) in DCM was used in the next synthetic step immediately.

[0313] Step (e): (l?)-2-((l-(7-methyl-2-morpholino-4-oxo-47 / -pyrido[l,2-a]pyrimidin-9- yl)ethyl)amino)benzoic acid (AZD6482)

[0314] Formula (Ila) Formula (la)

[0315] The compound of Formula (la) was prepared by base-mediated hydrolysis of the intermediate compound of Formula (Ila), conducted in methanol - aqueous sodium hydroxide mixture. After the reaction, methanol was distilled off, followed by a series of acid-base extractions with DCM. Solvent exchange to acetonitrile resulted in precipitation of the final product, which was isolated by filtration.

[0316] In a representative procedure, in a 5 L reactor, to a solution of the compound of Formula (II) in DCM taken directly from the previous synthetic step, MeOH (1.4 L, 5 rel. volumes) was added, and the mixture was distilled under reduced pressure to a residual volume of 1.4 L; the distillation was repeated three times with fresh portions of MeOH (2x 1.4 L, 2x5 rel. volumes), to a residual volume of 1.4 L. In a separate container, NaOH (74.7 g, 1.87 mol, 4.00 equiv.) solution in water (1.4 L, 5 rel. volumes) was prepared. The NaOH solution was charged to the reactor, and the mixture was heated at gentle reflux (internal temperature range +77 to +79 °C) for 1 h 17 min., at which point IPC confirmed full starting material conversion (compound of Formula (II) not detected). The reaction mixture was concentrated under reduced pressure to a residual volume of 1.4 L. The reaction mixture was then washed with DCM (1.4 L, 5 rel. volumes), and the DCM phase was discarded. To the remaining aqueous layer, a fresh portion of DCM (1.4 L, 5 rel. volumes) was added; with stirring, the mixture was neutralized with AcOH (224 g / 214 mL, 3.74 mol, 8.00 equiv.) to a pH of 4-5. The organic layer was drained and saved, while the remaining aqueous layer was extracted with a fresh portion of DCM (1.4 L, 5 rel. volumes). The organic extracts were combined, and the residual aqueous layer was discarded. The combined organic phase was charged back to the reactor and washed twice with water (2x 1.4 L, 2x5 rel. volumes). To the organic phase, MeCN (1.4 L, 5 rel. volumes) was added, and the mixture was distilled under reduced pressure to a residual volume of 2.8 L. Fresh portion of MeCN (1.4 L, 5 rel. volumes) was added, the mixture was seeded with a sample of the compound of Formula (I) (0.19 g, 0.1 mol%), and the distillation was continued to a residual volume of 2.8 L. A third portion of fresh MeCN (1.4 L, 5 rel. volumes) was added, and the distillation was continued to a residual volume of 2.8 L, at which point an off-white slurry formed. The slurry was held at 55 to 60 °C for 17 minutes, then cooled to 25 °C over 3 hours and stirred at that temperature for 3 days 14 hours.

[0317] The slurry was filtered, and the solids were washed with MeCN (2x0.6 L, 2x2 rel. volumes). The filter cake was dried under air suction for 23 hours.

[0318] Yield 125 g (65%), off-white solid.

[0319] Purity (HPLC, 265 nm): 99.4 %-area.

[0320] Enantiomeric purity (HPLC): 98.8 %-area (R).

[0321] Example 2: Optimisation of the synthesis of the compound of Formula (IVa)

[0322] It was identified by the present inventors that the compound of Formula (IVa) disclosed herein could be prepared from the compound of Formula (V) by asymmetric transfer hydrogenation in the presence of homogeneous ruthenium catalyst. It was determined that an optimum homogeneous ruthenium catalyst was RuCl[(S,S)-MsDpen](p-cymene), and that optimum conditions included carrying out the asymmetric transfer hydrogenation with a mixture of triethylamine and formic acid under reflux. Notably, after several reagent stoichiometry and mode of addition experiments, it was found that portionwise addition of formic acid to the reaction mixture provided faster reaction rates, limited impurity formation, and offered good reproducibility between different scale batches. Table 1 summarizes the representative batches manufactured under these conditions.

[0323] Table 1. Representative batches during optimization of the synthesis of the compound of formula (IV) aProduct / substrate / RRT 1.18 impurity ratio, HPLC, 265 nm.bExtra 0.05 eq formic acid was added and the mixture was stirred for 16 more hours.cThe amount of RRT 1.18 was not measured in IPC result.

[0324] Earlier experiments identified formation of a side product (or possibly a side product mixture) atRRT 1.18 at ca. 5-10 %-area levels (HPLC, 265 nm). Rejection of the impurity was poor during recrystallization, furthermore high recoveries of the product (compound of Formula (IV)) could not be achieved (the best conditions offered product recovery of 75% and enrichment from 91.3 to 95.9 %-area, HPLC analysis). Suppression of the side product formation via optimization of reaction conditions was highly desirable.

[0325] In the first optimization, the stoichiometry of formic acid / triethylamine (F / T mixture) was investigated. The first investigational experiment (SB087-50) was run on a 1.44 g (5 mmol) scale, with 2 equiv. of HCOOH and 10 equiv. of Et-N. After overnight stirring at RT, the product / substrate / impurity ratio was 62 / 29 / 9 %-area; the mixture was further heated at 40 °C for 1.5 hours, whereupon the aforementioned ratio was 87 / 1 / 12. After a solvent exchange to z- PrOAc, the product was isolated by filtration in a 1.08 g (75%) yield and 92 %-area purity (HPLC, 265 nm; RRT 1.18 content 8 %-area). The reaction proceeded smoothly overall, but the impurity levels were unsatisfactory, and a more detailed investigation was warranted.

[0326] In batch SB087-51, various loadings of both of HCOOH and EtsN were tested within the range of 1 to 3 molar equivalents in a 7-experiment matrix, as shown in Tables 2 and 3 below. The data suggested that excess formic acid not only promotes the impurity formation, but also reduced the reaction rate, as evidenced by lower starting material conversion at 17- hour mark. Analyzing the data at 38 h mark which included the product enantiomeric purity, best results were achieved when F / T ratio was kept close to 1:1. Table 2. SB087-51, IPC at 17h mark, Tj = 30 °C. Numbers (top to bottom) represent product / substrate / impurity ratio (HPLC %-area, 265 nm).

[0327] Table 3. SB087-51, IPC at 38 h mark, Tj = 45 °C. Numbers (top to bottom) represent product / substrate / impurity ratio (HPLC %-area, 265 nm) and enantiomeric purity.

[0328] Observing that the levels of RRT 1.18 side product decreased when less formic acid is used, an experiment with portionwise addition of HCOOH (SB087-53) was attempted, conducted on a 10 mol (2.9 g) scale. The experiment was sampled for IPC before each fresh portion of HCOOH was added. The results are shown in Table 4 below. The formation of the impurity could not be fully suppressed; nonetheless, the results were superior to the ones obtained in experiments SB087-51 earlier. Notably, stepwise addition of the reagent allowed the reaction to reach almost full substrate conversion in 3 hours.

[0329] Table 4. IPC of SB087-53 taken at 30 min intervals. Conditions: Tj = 45 °C.

[0330] As the RRT 1.18 impurity formation could not be completely avoided, a more selective isolation method was desired. Previous experiment (SB087-53) revealed that the solids which precipitate out as the reaction nears completion consisted of pure (>99 %-area) and highly enantioenriched (99.0% (S)) compound of Formula (IV) product, however if isolated as it was (only implementing brief cooling to 0 °C), the yield was only 50%.

[0331] The reaction with stepwise formic acid addition protocol was repeated on a larger 50- mmol (14-g) scale without issues (batch SB087-56), reaching completion within 3 h and with RRT 1.18 impurity detected at 7 % area in the reaction mixture. The reaction mixture was then sampled multiple times for isolation condition screening: the results are presented in Table 5.

[0332] Table 5. Compound of Formula (IV) isolation tests, batch SB087-56, 14.4 g (50 mmol) scale. Conditions: DCM (13 vol), EtsN (1.0 eq), FA added in portions (6><0.2 eq) 30 minutes apart, Ti = 40 °C.

[0333] In test 56-S1, solvent exchange from DCM to EtOH resulted in poorer quality solids. Dilution with MeCN (56-S2) offered improved enantioenrichment and purity, however the yield improvement was marginal; dilution with MTBE (56-S3) provided slightly more product at the expense of slightly lower optical purity. Concentration of the reaction mixture to ca. 1 / 2 original volume (56-S4) gave comparable results. Lastly, concentration to ca. 1 / 3 original volume followed by dilution with equal volume MeCN (56-S5) gave the highest yield in this set of experiments, however the obtained material was of comparatively poor optical purity.

[0334] Batch SB087-56-S2 conditions were selected for further scale-up, to prioritize optical purity over slightly higher product yields. Unfortunately, further optimization attempts for the 56-S2 conditions in batch SB087-58 failed to improve the yield and only reduced the isolated product’s optical purity - in fact, a later isolation attempt directly from the reaction mixture after 1 h cooling at 0 °C (batch SB087-65) produced improved results.

[0335] Advantageously, the designed conditions worked well during scale-up to 14-, 50-, and ultimately 359-g loading of Ketone- 1 (compound of Formula (V)) and were appropriate for multi-scale production of the compound of Formula (I).

[0336] It is noted that regarding mode of addition of formic acid, portionwise addition was chosen in this work based on process repeatability considerations, as addition of pre-measured reagent portions at fixed time intervals is easily documentable and reproducible. Addition of formic acid in a slow stream over a period of several hours also provided excellent results as shown in Table 6.

[0337] Table 6. Representative batches during further optimization of the synthesis of the compound of Formula (IV)

[0338] Example 3: Optimisation of the synthesis of the compound of Formula (Via)

[0339] It was identified by the present inventors that running the reaction with a slight excess of 4-nosylcholride (1.10 equiv.) in presence of pyridine (2.0 equiv.) in 8 relative volumes of MeCN afforded the compound of Formula (VI) in excellent yields and purity. This optimized process scaled without issues from 1.51 to 151 grams (10.0 mmol to 1.00 mol). A minor concern with a pronounced exotherm during reagent addition could be resolved with slow charging and dilution of methyl anthranilate with MeCN; no delayed exotherms were observed in verification batch production. On dilution with water prior to filtration, uniform, well-behaved slurries were obtained, no solid deposition on the reactor walls could be observed, and filtration rates were high on all scales. Notably, the product also dried well, despite being isolated from aqueous acetonitrile.

[0340] Example 4: Optimisation of the synthesis of the compound of Formula (Illa)

[0341] The present inventors found that a Mitsunobu reaction between a compound of Formula (IVa) as the alcohol and a compound of Formula (Via) as an N-nucleophile could be efficiently conducted on multi-gram scale. It was found that suitable conditions included triphenylphosphine (TPP) and diisopropyl azodicarboxylate (DIAD) in DCM. In an optimized procedure, once the reaction was complete, the solvent was exchanged to THF to precipitate a minor first crop of a racemic version of the compound of Formula (III) (rac-compound of Formula (Illa)), which was filtered and discarded. Subsequent solvent exchange to ethanol led to precipitation of the compound of Formula (Illa) (i.e., the desired (R) enantiomer) main crop, which was isolated by filtration. The identified reaction conditions worked well on both 28 g R&D scale, and on a 199 g production scale.

[0342] Table 7 Representative scale-up of the Mitsunobu reaction (step (c)).

[0343] * Issues not pertaining to the process design were encountered during Ba 1974 production due to a loss of solvents in the reactor. This resulted in the lower final yield.

[0344] The experiments defined in the example were focused on the product isolation. Based on the earlier-obtained solubility data for pure compound of Formula (Illa), it should exhibit very limited solubility in DCM or THF. Despite this reduced solubility, the product did not readily crystallize from the reaction mixture, likely due to PhsPO and DIAD-H2 byproducts.

[0345] In the initial isolation optimization experiment (SB087-46), the reaction mixture was diluted with THF (20 vol). A thin slurry formed and was filtered to obtain a 4% yield of the racemic material (‘crop 1’), similar to what was observed in earlier experiments. The filtrate was evaporated to dryness; to it, EtOH (50 vol) was added, causing precipitation - the mixture was heated to reflux until all solids dissolved, then cooled and filtered, obtaining a 69% yield of ‘crop 2’.

[0346] Considering that in the context of double precipitation, THF appeared to afford clean precipitation of the racemate, while DCM (reaction solvent) dissolved it to some degree, it was decided that dilution of reaction mixture was not optimal. Rather than using high volumes of THF (20 relative volumes or more) for dilution and precipitation as done in earlier experiments, a full solvent exchange to THF (10 volumes) was tested. For the second crystallization, the filtrate was solvent-swapped to ethanol (20 vol), and a brief reflux step was implemented, which greatly improved stirring and filtration of the obtained slurry. 43.6 g (80%) of the second crop was obtained in 99.6 %-area purity and ( ?)-enantiomer content of 96.1 %-area (confirmed in 2 independent deprotection experiments SB087-67 and -68). The lower optical purity in the above experiment was potentially attributed to extremely fine particle size of obtained rac-compound of Formula (Illa), leading to poor filtration with some of the material passing through the filter. In verification batch production, longer stirring time was implemented before racemate filtration for crystal maturation, as well as an addition of Seitz K200 clear-filtration aid. In the verification batch material, (7?)-enantiomer content of 98.7 %-area was obtained.

[0347] Overall, the developed conditions allowed isolation of the compound of Formula (Illa) on multi-gram scale in 77-80% yield and high enantiomeric purity of 98.7 %-area.

[0348] Example 5: Optimisation of the synthesis of the compound of Formula (Ila)

[0349] It was identified by the present inventors that the removal of the nosyl protecting group to obtain the compound of Formula (Ila) from the compound of Formula (Illa) could be advantageously effected on a multi-gram scale using a mixture of thioglycolic acid and tetramethylguanidine in acetonitrile. The inventors found that the most efficient method for carrying the produced compound of Formula (Ila) forward was to partition the resulting reaction mixture between water and DCM to remove water-soluble deprotection byproducts, obtain the product as a solution in DCM, and use it directly in the next step of the synthesis.

[0350] On smaller scale, the reaction had worked well under conditions of a thioglycolic acid - potassium carbonate mixture in wet DMSO. However, on a gram-scale the reaction was problematic for several reasons: DMSO was undesirable due to potential problems with drying; large excess of reagents was used; the reaction itself was heterogeneous; and isolation procedure resulted in a separation of sticky mass which then had to be stirred for an extended time period before a filterable slurry was obtained (the solids were also found to be contaminated with 3-10% mol / mol of the thioglycolic acid - nitrobenzene adduct that forms in the reaction). To determine the optimized representative procedure, the following conditions were tested.

[0351] Table 8. Step (d) development. All reactions: thioglycolic acid (2.5 equiv)

[0352] Thioglycolic acid with tetramethylguanidine (TMG) as a base in MeCN or acetone have been identified as potentially useful conditions. DMSO could thus be avoided, while the reaction itself is homogeneous for its entire duration. Between TMG and DBU which both worked well as bases, TMG was the preferred choice due to lower impurity formation in the reaction. In DCM, reactions proceeded well initially, but stalled at incomplete conversions, possibly due to cross-reactivity of thioglycolic acid with the solvent, resulting in reagent deactivation.

[0353] Under the HSCH2COOH / TMG conditions, thioglycolic acid loading could be optimized to 1.5 mol equiv. The Table below shows an optimization of reagent loading with stepwise addition of TMG, with best results obtained when HSCH2COOH / TMG ratio was maintained at ca. 1 :2, consistent with double deprotonation of thioglycolic acid being crucial for ensuring its reactivity. Table 9. Optimization of reagent loading. MeCN (10 vol), RT, 0.12-g scale. Attempts to isolate the product (the compound of Formula (Ila)) suggested that the compound of Formula (Ila) may be challenging to isolate as a crystalline solid, which led to development of process which involves carrying the compound of Formula (Ila) forward as an intermediate in a solution, without product isolation.

[0354] While developing the conditions for carrying the compound of Formula (Ila) forward as an intermediate in a solution, it was aimed to use a water-immiscible solvent, to implement an aqueous extraction for the water-soluble nosyl deprotection products in the reaction mixture (an ester hydrolysis test without prior removal of these byproducts gave rise to a complex mixture, making the washing step crucial).

[0355] MeTHF / MeCN 2: 1-v / v, z-PrOAc (both 10 vol dilution), z-PrOAc / MeCN 2: 1-v / v (15 vol dilution) were tested, but only resulted in heterogeneous reaction mixtures and poor conversions. DMSO (5 vol) was tested, but did not offer any benefit over MeCN, and isolation attempts with water dilution (similar to the original conditions) resulted in a poor product yield of 49%.

[0356] In the end, it was possible to run the reaction highly concentrated in MeCN (3-5 vol), and once complete, it could be diluted with MTBE or DCM and washed with water to remove all water-soluble nosyl deprotection byproducts. The solution of the compound of Formula (Ila) intermediate could then be solvent-swapped to MeOH for the subsequent ester hydrolysis reaction.

[0357] Example 6: Optimisation of the synthesis of the compound of Formula (la)

[0358] It was identified by the present inventors that the compound of Formula (la) could be synthesized by exchanging the solvent of a solution of the compound of Formula (Ila) directly from the previous synthetic to methanol, and submitting the compound to basic hydrolysis in methanol - aqueous sodium hydroxide mixture. After the reaction, methanol was distilled off, followed by a series of acid-base extractions with DCM. Solvent exchange to acetonitrile precipitated the final product, which was isolated by filtration.

[0359] While the hydrolysis reaction proceeded cleanly, the isolation of the compound of Formula (la) required investigation, including the following conditions that were trialled on a 10-mmol scale:

[0360] (l) The volatiles (MeOH) were distilled off. The remaining basic aqueous solution of the compound of Formula (la) was washed with DCM to remove possible organic impurities, then acidified to pH 4 5 and extracted with DCM, pulling the product into the organic phase. Solvent exchange to MeCN then cleanly precipitated the product.

[0361] (2) The reaction mixture was directly acidified to pH 4-5 and heated at reflux for 15 min to afford the product slurry. Acidification at room temperature resulted in oiling out of the product which coalesced into semi-solid gum; reflux was necessary for proper crystallization of the solids.

[0362] In both experiments, materials of 99.2 %-area purity by HPLC were obtained, however due to uncontrolled oiling out and solidification of the material in experiment (2), these conditions were deemed less robust than the acid-base extraction approach of experiment (1), which were chosen to use in further scale-up.

[0363] Solvent choice for precipitation was based on the preliminary solubility assessment. In the 1st campaign, the final product (the compound of Formula (la)) was isolated by evaporation to dryness, yielding what likely was an amorphous solid, with HPLC purity of 97.6 %-area. Trituration of the material was tested in a series of solvents, summarized in the Table below. Among these, several solvents (MeCN, z-PrOAc, z-PrOH, acetone) caused clean white slurry formation from an initially yellow solid, suggesting crystallization. In later development, it became apparent that MeCN causes clean product precipitation regardless of impurity content.

[0364] Table 10. Preliminary solubility of a likely amorphous sample of the compound of Formula

[0365] (la) aNumbers given where full dissolution could be achieved, NA - no full dissolution in the specified solvent volume.

[0366] Overall, in optimizing the procedure for steps (e) and (d), the inventors developed a process for efficient and scalable nosyl deprotection and ester hydrolysis, which was successfully scaled up to 284 g loading of the compound of Formula (III) and provided a >99 %-area (HPLC) pure product (the compound of Formula (I)) in up to 65% yield over two synthetic steps.

[0367] Example 7: Recrystallisation of the compound of Formula (la)

[0368] Formula (la)

[0369] It was identified by the present inventors that purified samples of the compound of Formula (la) could be obtained by clear-filtration of a crude solution of the compound of Formula (la) in DCM, followed by solvent exchange to isopropanol, leading to crystallization of the product. The material was isolated by filtration.

[0370] In a representative procedure, to a 5-L reactor, DCM (1.2 L, 10 rel. volumes) was charged. Crude compound of Formula (la) (120 g, 0.294 mol, 1.00 equiv.) was then charged, and the mixture was heated at gentle reflux (39 to 40 °C internal temperature) for 30 minutes, until the reactor walls and moving parts were free of solid residue. The resulting thin slurry (consisting primarily of rac -compound of Formula (la) and inorganic impurities) was cooled to 25 °C and clear-filtered (Seitz K200 filter aid); filter was rinsed with DCM (0.12 L, 1 rel. volume), and the filtrate was charged back to the reactor. z-PrOH (1.2 L, 10 rel. volumes) was charged to the reactor, and the mixture was distilled under reduced pressure to a residual volume of 1.2 L. A fresh portion of z-PrOH (1.2 L, 10 rel. volumes) was added, and distillation was continued to a residual volume of 1.2 L, at which point a white slurry had formed. The reactor was re-pressurized with nitrogen to atmospheric pressure, and the mixture was heated at gentle reflux (80 to 83 °C internal temperature) for 43 minutes until the reactor walls were free of solids (manual dislodging was necessary). The mixture was then cooled to 25 °C over 2 hours, held at 25 °C for 20 minutes, and filtered. The filter cake was washed with 2 portions of z-PrOH (2x0.24 L, 2x2 rel. volumes) and dried under air suction for 15 hours.

[0371] Yield: 90.7 g (77%), off-white solid.

[0372] LOD: 0.6 %-w / w.

[0373] Purity (HPLC, 265 nm): 99.8 %-area.

[0374] Enantiomeric purity (HPLC): 99.8 %-area (R).

[0375] Assay by qNMR: 93.0 %-w / w.

[0376] Water content: 0.1 %-w / w.

[0377] Sulphated ash: 0.1 %-w / w.

[0378] Ru content (ICP-MS): <0.01 ppm: Since a ruthenium catalyst was used in Step (a) of the synthesis, ruthenium quantification by ICP MS was included in the verification of the final product before and after recrystallization. Neither samples contained ruthenium above limit of detection (Ru < 0.01 ppm).

[0379] During development of the representative procedure, aqueous ethanol mixtures was chosen as the starting solvent. However, it was found that crude aqueous ethanol solutions of the compound of Formula (la) were unstable and tended to oil out uncontrollably, while required solvent quantities were large and material recoveries poor. Variations in solubility between different batches of the crude material were also observed. Additionally, solvents such as MeCN-water, MeTHF, z-PrOH were tested, however the crude product did not dissolve in these solvents even after prolonged reflux. These preliminary screening results are summarized in Table 11 below.

[0380] Table 11. Initial recrystallization experiments using 500 mg samples of the compound of Formula (la)

[0381] In sub-batch SB087-75-2, considerable amount of precipitate formed in the filtrate as EtOH evaporated off - this precipitate was also filtered and analysed. Notably, the undesired enantiomer concentrated in the first product crop, maintaining the trend of the racemate being significantly less soluble than the pure enantiomer, as observed with other intermediates; the second crop was notably enantioenriched, with only 0.13% of the undesired (^-enantiomer detected. In the sub-batch SB087-75-3, only the near-racemic product collected on the filter, while the remaining extremely fine particles passed through. As with 75-2, more crystals precipitated in the filtrate, confirming that the conditions are not suitable. Furthermore, in all recrystallization runs employing water as antisolvent, oiling-out of the liquors and amorphous film formation on most surfaces in contact with the liquors was observed, likely caused by evaporation of the organic phase - that is, aqueous systems were inherently unstable, and an alternative was necessary. During solvent screening it was noticed that DCM dissolved the crude compound extremely well, leaving behind a small amount of extremely fine precipitate; assuming that the precipitate might be the poorly soluble racemic material it was attempted to clear-filter the suspension, and then isolate the product by solvent exchange (batch SB087-78 experiments, see Table 12), testing ethanol, isopropanol, and aqueous ethanol as antisolvents. Since the isopropanol conditions provided superior product recovery, these were the conditions used in the verification batch manufacture. In the verification batch, the same conditions with extended stirring times and employment of Seitz K200 clear-filtration aid have afforded the final product in a high enantiomeric purity of 99.8 %-w / w of the desired (7?)-enantiomer. Table 12. Further optimisation of recrystallization conditions.

[0382] An X-ray powder diffraction (XRPD) pattern of a sample of the compound of Formula (la) prepared as described above is illustrated in Figure 1. Absolute configuration of the chiral center in compound of Formula (la) prepared as described above was determined by single crystal X-ray diffraction. Single-crystal diffraction data were collected on an XtaLAB Synergy-S Dualflex diffractometer (Rigaku Corporation, Tokyo, Japan) equipped with a HyPix6000 detector and micro-focus sealed X- ray tube using Cu Ka radiation (X = 1.54184 A). Single crystal was fixed with oil in a nylon loop of a magnetic CryoCap and set on a goniometer head. The samples were cooled down to 150 K, and co-scans were performed with a step size of 0.5°. Data collection and reduction were performed with the CrysAlisPro 1.171.40.35a software (Oxford Diffraction Ltd., Abingdon, UK). Structure solution and refinement were performed with SHELXL and SHELXT software that are parts of the CrysAlisPro and Olex2 suites.

[0383] Figure 2 shows an ORTEP view of a sample of the compound of Formula (la) prepared as described above, demonstrating the absolute configuration at the center of chirality to be (R).

[0384] The absolute configuration was determined by the Flack method using anomalous dispersion of oxygen atoms. The Flack parameter is calculated by the refinement program SHELXL. For correct absolute configuration the Flack parameter assumes the value close to 0.0. If configuration is wrong, the Flack parameter is close to 1.0. For compound of Formula (I) structure the values of Flack parameter were: 0.046(311) by classical fit to all intensities and 0.087(105) from 1360 selected quotients (Parsons' method). The values obtained were close to zero and, therefore, indicate that determined and shown in Figure 2 absolute configuration is correct and chiral center at carbon atom has R-configuration. For reliability, the Flack parameter was also computed for inverted structure and obtained the following values: 0.924(312) by classical fit to all intensities and 0.909(105) from 1359 selected quotients (Parsons' method). Since, in this case, the values were close to 1.0, the inverted structure had wrong absolute configuration. Details of data collection and structure refinement are provided in Tables 13, 14 and 15.

[0385] Table 13, Crystal data Table 14. Data collection

[0386] Table 15. Refinement Geometric parameters: not shown.

[0387] Further optimization experiments focused on crystallisation procedures for generating pre-production scales of the product, e g. 15 g and higher. The following optimization experiments were conducted using 5 g of crude amorphous solids of compound of Formula (la). Table 16. Optimization of crystallization conditions on multi-gram scale

[0388] Most attempted conditions resulted in various degrees of solid deposition on flask walls.

[0389] As the issue became more pronounced on a larger scale, these conditions were not tested any further.

[0390] It was noted that acid-base processes (experiments 5 and 6) in which the material was dissolved in aqueous ethanol with base addition, clear-filtered and neutralized with AcOH, resulted in clean precipitation with no solid deposition, however the precipitation was too rapid and produced solids which stirred, filtered and dried poorly, and on drying formed hard pieces of material that would likely require additional processing such as milling. When attempted to repeat the experiment, the product oiled out during neutralization, and the precipitate was not homogeneous.

[0391] Dissolving the material in ethanol under reflux followed by addition of seed suspension in water at 60 °C resulted in clean precipitation of the product with no observed solid deposition on the flask walls. The formed slurry stirred, filtered, and dried well. This process was investigated further (experiments SB087-110) with variations to ethanol-water ratio and isolation temperature. The results are shown in Table 17.

[0392] Table 17. Optimization of crystallization conditions on multi-gram scale Higher water content resulted in slightly higher isolated yields; between experiments 1 and 2, assay values and HPLC purities of isolated products were comparable, yet visually the material from experiment 2 appeared slightly more colored. Higher ethanol content produced the highest assay material with most of the colored impurities remaining in the filtrate, however the yield was also significantly lower. Overall the product slurries in all experiments were well-behaved, and no solids deposited on flask walls at any moment, indicating that the conditions were suitable for crystallization of the product (compound of Formula (I)) on at least a 5 g scale.

[0393] Example 8: Synthesis of the compound of Formula (la)

[0394] A process for manufacture of the compound of Formula (la), via the intermediate compound of Formula (IVa), has been optimised and tested in the production of technical / process verification batch with a final product output of 100 g and overall yield of 20% in 5 synthetic steps (4 steps longest linear sequence), including final product recrystallization.

[0395] Individual steps of the synthetic route and synthesis of the intermediate compounds disclosed herein were as follows and are summarised in Figure 5.

[0396] Step (a): ( )-9-(l-Hydroxyethyl)-7-methyl-2-morpholino-4H-pyrido[l,2-a]pyrimidin-4- one

[0397] The compound of Formula (Va) may be prepared, for example, as described in W02009 / 093972 and / or W02004 / 016607 or purchased from a commercial supplier.

[0398] In a representative procedure, to a clean 5 L container the compound of Formula (Va) (431 g, 1.50 mol, 1.00 equiv ), was charged, followed by DCM (4.3 L, 10 rel. volumes). The mixture was clear-filtered on a fritted glass filter (porosity 3) equipped with a Seitz K-200 filter aid. The undissolved compound of Formula (Va) residue on the filter was solubilized with additional DCM (900 mL, 2 rel volumes) and filtered Both filtrates were combined. A clear yellow filtrate was obtained, with a layer of fine brown particles trapped on the filter. The solids were discarded.

[0399] The filtrate was transferred to a 5 L reactor. Et3N (152 g / 210 mL, 1.50 mol, 1.00 equiv.) andRuCl[(S,S)-MsDpen](p-cymene) (4.2 g, 0.00750 mol, 0.5 mol%, FountainBridgeLtd., UK) were charged, and the reactor was evacuated and refilled with nitrogen three times. The reactor contents were then brought to a gentle reflux. A ca. 5M solution of formic acid (82.9 g / 67.9 mL, 1.80 mol, 1.20 equiv.) in DCM (274 mL, 0.63 rel. volumes) was prepared, and then charged to the reaction mixture in six equal portions each spaced 30 minutes; between the addition of 5th and 6th portions, a product slurry started to form. After the addition was finished, the reaction mixture was held at reflux for an additional 60 minutes. IPC confirmed full starting material conversion (0.6 %-area Ketone-1, HPLC, 265 nm, IPC limit Ketone-1 < 3.0 %-area).

[0400] The reaction mixture was cooled to 0 °C, and then held at that temperature for 16 hours. The product slurry was filtered and washed with DCM (0.43 L, 1 rel. volume), DCM-MTBE 1 : 1 v / v (0.43 L, 1 rel. volume), and MTBE (0.43 mL, 1 rel. volume), in that order. The product was dried on the filter for 73 hours.

[0401] Yield: 243 g (56%), off-white solid.

[0402] LOD: 0.6 %-w / w.

[0403] Purity (HPLC, 265 nm): 99.1 %-area.

[0404] Enantiomeric purity (HPLC): 97.3 %-area (5).

[0405] Step (b): Methyl 2-((4-nitrophenyl)sulfonamido)benzoate

[0406] Methyl anthranilate 4-Nosylchloride Formula (Via)

[0407] In a representative procedure, to a 5-L reactor, MeCN (0.90 L, 6 rel. volumes) was charged, followed by 4-nosylchloride (244 g, 1.10 mol, 1.10 equiv.) and pyridine (158 g / 161 mL, 2.00 mol, 2.00 equiv.). A solution of methyl anthranilate (151 g / 129 mL, 1.00 mol, 1.00 equiv.) in MeCN (0.30 L, 2 rel. volumes) was prepared and charged to the reactor over 20 minutes, maintaining the internal temperature below 35 °C. The mixture was stirred at 25 °C for an additional 4 hours. IPC sample was taken, which showed incomplete starting material conversion (8.9 %-area methyl anthranilate, HPLC, 265 nm, IPC limit < 2.0 %-area). The reaction mixture was stirred for another 1 hour and a second IPC sample was taken, which contained 3.7 %-area methyl anthranilate. Based on the earlier-obtained evidence that the excess starting material is rejected during isolation, it was decided to continue with the workup and product isolation.

[0408] Water (1.2 L, 8 rel. volumes) was added to the reaction mixture, and the reactor contents were stirred for an additional hour. The mixture was then filtered, and the filter contents were washed twice with MeCN-water 1 : 1-v / v (2x0.30 L, 2x2 rel. volumes), then dried under suction for 16 hours.

[0409] Yield: 305 g (91%), light-yellow solid.

[0410] LOD: 0.6 %-w / w.

[0411] Purity (HPLC, 265 nm): 99.8 %-area.

[0412] Step (c): Methyl (!?)-2-((iV-(l-(7-methyI-2-morpholino-4-oxo-4H-pyrido[l,2-a]pyrimidin-

[0413] 9-yl)ethyl)-4-nitrophenyl)sulfonamido)benzoate

[0414] In a representative procedure, to a 5-L reactor, DCM (1.2 L. 5 rel. volumes) was charged, followed by (S)HRIOOl-Ol (238 g, 0.825 mol, 1.00 equiv), HRI001-03 (305 g, 0.908 mol, 1.10 equiv.), and TPP (325 g, 1.24 mol, 1.50 equiv.). The reactor contents were cooled to -5 °C, and DIAD (251 g / 244 mL, 1.24 mol, 1.50 equiv.) was charged over 59 minutes, without letting the internal temperature exceed 0 °C. The reaction mixture was held at 0 °C for an additional 77 minutes, warmed to 25 °C over 3 hours, then stirred at +20 to +25 °C for 13 hours. IPC confirmed full starting material conversion ((S)HRI001-01 0.9 %-area, HPLC, 265 nm, IPC limit < 1.0 %-area).

[0415] The reaction mixture was diluted with THF (2.4 L, 10 rel. volumes), then distilled under reduced pressure to a total volume of 3.3 L. The mixture was cooled to +20 °C and held at that temperature for 90 minutes, then further cooled to +5 °C and held at that temperature for 3 hours, during which a thin slurry of rac-HRIOO 1 -04 formed. The slurry was held at 10 °C for 16 hours and then filtered on a fritted glass filter equipped with Seitz K200 filter aid; the solids were rinsed with additional THF (0.24 L, 1 rel. volume) and discarded. The reactor was cleaned, and the filtrate was transferred back to the reactor. EtOH (1.9 L, 8 rel. volumes) was added, and the solvents were distilled off under atmospheric pressure to a total volume of 2.4 L; distillation was repeated with another portion of EtOH (1.9 L, 8 rel. volumes) to the end volume of 3.4 L. The mixture was cooled to from +70 to +20 °C over 300 min (10 °C / h; crystallization initiated at Ti = 63 °C) and held at that temperature for 13 hours. The slurry was filtered; the solids were washed with EtOH (2x0.48 L, 2x2 rel. volumes) and dried under air suction for 69 hours. Yield: 375 g (75%), yellow solid.

[0416] LOD: 0.5 %-w / w.

[0417] HPLC purity: 99.0 %-area.

[0418] Enantiomeric purity (HPLC): 99.9 %-area (A).

[0419] Step (d): Methyl R)-2-((l-(7-methyl-2-morpholino-4-oxo-4 / / -pyrido[l,2-a]pyrimidin-9- yl)ethyl)amino)benzoate

[0420] In a representative procedure, to a 5-L reactor, MeCN (1.7 L, 5 rel. volumes) was charged, followed by (R)HRI001-04 (338 g, 0.556 mol, 1.00 equiv.) and thioglycolic acid (76.9 g / 58.2 mL, 0.835 mol, 1.50 equiv.). To the resulting suspension, thioglycolic acid (192 g / 209 mL, 1.67 mol, 3.00 equiv.) was added slowly over 15 minutes, maintaining internal temperature below 37 °C. After the addition was finished, the mixture was stirred at 25 °C for 17 hours. IPC1 showed incomplete starting material conversion (3.5 %-area (R)HRI001-04). Additional portions of thioglycolic acid (7.7 g / 5.8 mL, 0.084 mol, 0.15 equiv.) and thioglycolic acid (19.2 g / 20.9 mL, 0.167 mol, 0.30 equiv.) and stirring was continued for another 23 hours, at which point ZPC2 confirmed full conversion (< 0.1 %-area (R)HRI001-04).

[0421] MTBE (1.7 L, 5 rel. volumes) was charged to the reactor, followed by water (1.7 L, 5 rel. volumes), and another portion of MTBE (1.7 L, 5 rel. volumes). The reactor contents were then washed water (3x 1.7 L, 3x5 rel. volumes). Aqueous washings were discarded. The obtained (R)HRI001-02 solution in MTBE was used in the next synthetic step immediately. Step (e): ( / ?)-2-((l-(7-methyl-2-morpholino-4-oxo-4H-pyrido[l,2-«|pyrimidin-9- yl)ethyl)amino)benzoic acid (AZD6482)

[0422] In a representative procedure, in a 5 L reactor, to a solution of (R)HRI001-02 in MTBE taken directly from the previous synthetic step, MeOH (1.7 L, 5 rel. volumes) was added, and the mixture was distilled under reduced pressure to a residual volume of 1.7 L; the distillation was repeated another time with a fresh portion of MeOH (1.7 L, 5 rel. volumes), to a residual volume of 1.7 L. In a separate container, NaOH (89.0 g, 2.23 mol, 4.00 equiv.) solution in water (1.7 L, 5 rel. volumes) was prepared. The NaOH solution was charged to the reactor, and the mixture was heated at gentle reflux (internal temperature range +77 to +79 °C) for 1 h 15 min., at which point IPC confirmed full starting material conversion ((R)HRI001-02 not detected).

[0423] The reaction mixture was concentrated under reduced pressure to a residual volume of 1.7 L. DCM (1.7 L, 5 rel. volumes) was added, forming a biphasic mixture; with strong stirring, the mixture was neutralized with AcOH (267 g / 254 mL, 4.45 mol, 8.00 equiv.) to a pH of 4- 5. The organic layer was drained and saved, while the remaining aqueous layer was extracted with a fresh portion of DCM (1.7 L, 5 rel. volumes). The organic extracts were combined, and the residual aqueous layer was discarded.

[0424] The combined organic phase was charged back to the reactor and washed twice with water (2x 1.7 L, 2x5 rel. volumes). To the organic phase, MeCN (1.7 L, 5 rel. volumes) was added, and the mixture was distilled under atmospheric pressure to a residual volume of 3.4 L. Fresh portion of MeCN (1.7 L, 5 rel. volumes) was added, and the distillation was continued to a residual volume of 3.4 L, at which point an off-white slurry formed. A third portion of fresh MeCN (1.7 L, 5 rel. volumes) was added, and the distillation was continued to a residual volume of 3.4 L. The slurry was cooled from +80 °C to +20 °C over 6 hours (10 °C / h) and stirred at that temperature for 10 hours.

[0425] The slurry was filtered, and the solids were washed with MeCN (2x0.7 L, 2x2 rel. volumes). The filter cake was dried under air suction for 2 hours, at which point a constant weight of the filter cake was confirmed. Yield 136 g (60%), off-white solid.

[0426] LOD: 0.7 %-w / w.

[0427] Purity (HPLC, 265 nm): 99.3 %-area.

[0428] Enantiomeric purity (HPLC): > 99.9 %-area (R).

[0429] Step (f): Recrystallization of ( / ?)-2-(( l-(7-niethyl-2-niorpholino-4-oxo-4 / / -pyrido|l,2- a]pyrimidin-9-yl)ethyl)amino)benzoic acid (AZD6482)

[0430] In a representative procedure, to a 2-L round-bottom flask, DCM (1.26 L, 10 rel. volumes) was charged. Crude compound of Formula (la) (126 g, 0.326 mol, 1.00 equiv.) was then charged, and the mixture was heated at gentle reflux (39 to 40 °C internal temperature) for 10 minutes, until the reactor walls and moving parts were free of solid residue. The resulting turbid solution (consisting primarily of rac-HRIOOl and inorganic impurities) was cooled to 25 °C and clear-filtered (Seitz K-200 filter aid); filter was rinsed with DCM (0.13 L, 1 rel. volume), and the filtrate was charged to a 5 L reactor.

[0431] EtOH (1.26 L, 10 rel. volumes) was charged to the reactor, and the mixture was distilled under reduced pressure to a residual volume of 1.3 L. A fresh portion of EtOH (1.26 L, 10 rel. volumes) was added, and distillation was continued to a residual volume of 2.0 L, at which point some crust formation started on the reactor walls and eventually a slurry started forming. Water (0.63 L, 5 rel. volumes) was added, and distillation was continued to a residual volume of 2.0 L. The reactor was re-pressurized with nitrogen to atmospheric pressure, and the mixture was heated at gentle reflux (80 to 83 °C internal temperature) for 30 minutes until the reactor walls were free of solids.

[0432] The mixture was cooled to +70 °C . In a separate container, a seed sample of a compound of Formula (la) (630 mg, 0.5 %-w / w) was weighed in and suspended in a mixture of water (0.25 L, 2 rel. volumes) and EtOH (0. 13 L, 1 rel. volume). The seed slurry was charged to the reactor over the duration of 30 minutes while maintaining the internal temperature between +60 and +65 °C. After the seeding, another portion of water (0.25 L, 2 rel. volumes) was added over 30 minutes at the same temperature. A 10 °C / h cooling ramp was set up, and the mixture was cooled to a 0 °C endpoint (7 h), then held at that temperature for 11 hours.

[0433] The slurry was filtered, and the solids were washed with a 3:2-v / v EtOH-water mixture (2*2.5 rel. volumes). The filter cake was dried under air suction for 24 h.

[0434] A representative XRPD spectrum, IR spectrum and DSC thermogram are shown in Figures 1, 3 and 4, respectively.

[0435] Yield: 100 g (79%), off-white solid. LOD: 0.4 %-w / w.

[0436] Purity (HPLC, 265 nm): 99.7 %-area.

[0437] Enantiomeric purity (HPLC): >99.97 %-area (R).

[0438] Water content: 0.03 %-w / w. Sulphated ash: 0.1 %-w / w.

[0439] Ru content (ICP-MS): < 0.01 ppm.

[0440] A 700-gram scale GMP production has been successfully carried out based on this process, starting from 2.80 kg Ketone- 1 raw material, and obtaining 0.593 kg of the final (R)HRI001 product, in an overall yield of 15%. Due to technical difficulties encountered in

[0441] Step 3 of the production, part of the material was lost (some advanced intermediates are stored and sustain GMP status), and the isolated yield was lower than planned. Despite this, the final product amount was only 15% off the 700-g target. A summary of the manufacturing steps and isolated yields is provided in Table 19: Table 19. A summary of the manufacturing steps and isolated yields

Claims

Claims:

1. A process for preparing an intermediate compound of Formula (IV) comprising: asymmetric reduction of the ketone of Formula (V) to form a compound of Formula (IV).) wherein R1is Ci-6 alkyl.

2. The process of claim 1, wherein a ruthenium-catalyst is used for asymmetric reduction of the ketone.

3. The process of claim 2, wherein the ruthenium-catalyst is RuCl[(S,S)-MsDpen](p- cymene).

4. The process of any one of claims 1 to 3, wherein a hydrogen source for the asymmetric reduction comprises triethylamine and formic acid.

5. The process of claim 4, wherein formic acid and triethylamine is used in a formic acid: tri ethylamine ratio of between about 5: 1 and 1:5.

6. The process of claim 5, wherein formic acid and triethylamine is used in a formic acid: triethylamine ratio of between about 1.2: 1 and 1:1.2, or about 1:1.

7. The process of any one of claims 1 to 6, wherein formic acid is added portionwise to a solution comprising the compound of Formula (V) and ruthenium-catalyst in solvent.

8. The process of claim 7, wherein about 0.2 molar equivalents of formic acid are added every 30 minutes over three hours.

9. The process of any one of claims 1 to 8, wherein the asymmetric reduction is performed in the presence of a solvent, wherein the solvent is dichloromethane.

10. The process of any one of claims 1 to 9, wherein the process provides a compound of formula (IV) having an enantiomeric purity of at least 95%, or at least 97%.

11. The process of any one of claims 1 to 10, wherein R1is CHs.

12. A process for producing a compound of Formula (I):Formula (I) the process comprising: a) converting a compound of Formula (V) to a compound of Formula (IV) according to the process of any one of claims 1 to 11; b) a Mitsunobu reaction of the compound of Formula (IV) with a compound of formula(VI) to form a compound of Formula (III):c) converting the compound of Formula (III) to form the compound of Formula (I)R1is an optionally substituted Ci-6 alkyl or an optionally substituted C3-7 cycloalkyl; andR2is an optionally substituted C1-6 alkyl or an optionally substituted C3-7 cycloalkyl.

13. The process of claim 12, wherein the Mitsunobu reaction is performed in the presence of triphenylphosphine (TPP) and an azodicarboxylate.

14. The process of claim 13, wherein the azodi carb oxy late is selected from diethyl azodicarboxylate (DEAD) or diisopropyl azodicarboxylate (DIAD).

15. The process of any one of claims 12 to 14, wherein the Mitsunobu reaction is performed in the presence of a solvent.

16. The process of claim 15, wherein the aprotic solvent or non-polar solvent is selected from dichloromethane (DCM), tetrahydrofuran (THF), methyltetrahydrofuran (MeTHF), acetonitrile (MeCN), V-m ethyl pyrrolidone (NMP), pyridine, toluene, hexanes, n-heptane, ethyl acetate (EtOAc), methylisopropyl ketone (MIPK), 7V,V-dimethylformamide (DMF), dimethylsulfoxide (DMSO), or any combinations thereof.

17. The process of any one of claims 12 to 16, wherein the compound of formula (III) is isolated prior to use in step (c), wherein the isolation comprises: solvent exchange into THF; filtration to form a filtrate; solvent exchange of the filtrate into an alcohol to precipitate the compound of formula (III); and isolation of the precipitate.

18. The process of claim 17, wherein the alcohol is selected from methanol, ethanol and isopropyl alcohol, or any combinations thereof.

19. The process of claim 18, wherein the alcohol is ethanol.

20. The process of any one of claims 12 to 19, wherein the compound of Formula (VI) is prepared by a process comprising sulfonylation of an anthranilate of Formula (VII) using a nosylating agent:Formula (VII) Formula (VI)whereinwherein, R2is Ci-6 alkyl.

21. The process of claim 20, wherein the nosylating agent is 4-nosylchloride.

22. The process of claim 20 or claim 21, wherein the anthranilate of Formula (VII) is methyl anthranilate.

23. The process of any one of claims 20 to 22, wherein the nosylating agent is provided in molar equivalents of between about 0.01 and 5, between about 0.05 and 4, or between about 0.1 and 2, or between about 0.5 to 1.5, or about 1.1 relative to the anthranilate.

24. The process of any one of claims 20 to 23, wherein a base reagent for sulfonylation of the anthranilate of Formula (VII) comprises pyridine.

25. The process of any one of claims 20 to 24, wherein a base reagent for sulfonylation of the anthranilate of Formula (VII) does not comprise DMAP.

26. The process of any one of claims 20 to 25, wherein the sulfonylation reaction is performed in the presence of a solvent and wherein the amount of solvent is present in volume equivalents (L), relative to the molar amount of the compound of the anthranilate of Formula (VII), of between about 1 and 15, between about 5 and 10, or between about 6 and 8.

27. The process of claim 26, wherein the solvent is DCM.

28. The process according to any one of claims 12 to 27, wherein step c) comprises: cl) de-nosylating the compound of Formula (III) to form a compound of Formula (II):c2) treating the compound of Formula (II) with a base to form a compound of FormulaFormula (II) Formula (I)29 The process of claim 28, wherein de-nosylating comprises reacting the compound ofFormula (III) with thioglycolic acid in the presence of a base.

30. The process of claim 29, wherein the base is tetramethylguanidine.

31. The process of claim 30, wherein the base used in step c2) is NaOH.

32. The process of any one of claims 28 to 31, wherein the compounds are reacted in situ, without isolation, in the consequent reaction.

33. The process of any one of claims 12 to 32, further comprising recrystallising the compound of Formula (I) using iso-propanol.

34. The process of any one of claims 12 to 33, wherein R1is CHa, R2is CHa and Ns is35. The process of any one of claims 12 to 34, wherein the process provides a compound of formula (I) having an enantiomeric purity of at least 95%, or at least 97%.

36. An enantiomerically pure compound of Formula (IV)wherein Ri is Ci-6 alkyl.

37. An enantiomerically pure compound of Formula (IVa)38. An enantiomerically pure compound of Formula (III)Formula (III) whereinR1and R2are independently Ci-6 alkyl, and39. An enantiomerically pure compound of Formula (Illa)wherein40. A compound of Formula (I) prepared by the process of any one of claims 12 to 35.

41. The compound of any one of claims 35 to 40, wherein the compound has an enantiomeric purity of at least 95%, or at least 97%.

42. A process for preparing AZD6482 starting from a compound of Formula (IVa).

43. The process of claim 42, wherein AZD6482 has an enantiomeric purity of at least 95%, or at least 97%.

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