EA043669B1 - PURINONE COMPOUNDS AND THEIR USE IN CANCER TREATMENT - Google Patents

Description

Field of technology

The present description generally relates to substituted purinone compounds and their pharmaceutically acceptable salts. These compounds and their pharmaceutically acceptable salts selectively modulate DNA-dependent protein kinase (DNA-PK), and therefore, the present disclosure also relates to the use of such compounds and their salts for the treatment or prevention of DNA-PK-mediated disease, including cancer. The present description further relates to crystalline forms of purinone compounds and their pharmaceutically acceptable salts; pharmaceutical compositions containing such compounds and salts; kits containing such compounds and salts; methods for preparing such compounds and salts; intermediates useful in the preparation of such compounds and salts; and methods of treating DNA-PK mediated disease, including cancer, using such compounds and salts.

State of the art

DNA-PK is a nuclear serine-threonine protein kinase complex consisting of the catalytic subunit DNA-PKcs and a heterodimer of Ku proteins (Ku70/Ku80). DNA-PK plays a critical role in the repair of DNA double-strand breaks (DSBs), which serves to maintain genome integrity, and in the process of V(D)J recombination, which results in the highly diverse repertoire of antibodies/immunoglobulins and T-cell receptors found on B and T cells, respectively. DNA-PK has also been linked to a variety of other biological processes, including modulation of chromatin structure, telomere length maintenance, transcriptional regulation, and the response to replicative stress (Smith and Jackson, 1999; Goodwin and Knudsen, 2014).

DNA DSBs are considered the most lethal forms of damage that can occur in a cell. To counteract the serious threats posed by DNA DSBs, eukaryotic cells have evolved several mechanisms to mediate their repair. In higher eukaryotes, the predominant mechanism is non-homologous DNA end joining (NHEJ). It is an error-prone DSB repair pathway involving direct ligation of broken DSB ends that occurs during all phases of the cell cycle, and is primarily used during early G1/S phases where the sister chromatid is not yet available to serve as a template (Hartlerode and Scully, 2009). This is in contrast to the second major DSB repair pathway, homologous recombination (HR), which occurs primarily during the G2/M phases of the cell cycle when intact sister chromatids are available (San Filippo et al., 2008). Other mechanisms driving the selection of NHEJ or HR for DSB repair are not fully defined, although blunt, minimally processed DNA ends are repaired by NHEJ, whereas 3' end truncation is required for HR to occur (Symington and Gautier, 2011). End truncation is controlled by the interaction of BRCA1 and 53BP1, with 53BP1 mediating NHEJ passage by suppressing end truncation (Escribano-Diaz et al., 2013).

NHEJ is initiated through the recognition and binding of broken DNA ends by the ring-shaped Ku70/Ku80 heterodimer, followed by the recruitment of DNA-PKcs through its interaction with Ku and DNA. Recruitment of DNA-PKcs facilitates the movement of the Ku heterodimer into the DNA duplex, which allows DNA-PKcs to act as a binding element for broken DNA ends and prevent destruction by exonucleases (Yoo and Dynan, 1999). Binding to DNA promotes activation of the catalytic activity of DNA-PKcs. Arguably, the most important DNA-PK substrate is the kinase subunit itself, since autophosphorylation is critical for regulating DNA end processing, enzyme inactivation, and complex dissociation (Chan et al., 2002). The most well-studied autophosphorylation sites are Ser2056 and Thr2609 (Douglas et al., 2002). DNA-PKcs phosphorylates and modifies the activity of multiple substrates that mediate NHEJ, including Artemis, Ku70, Ku80, and DNA ligase 4 (Neal and Meek, 2011); it also phosphorylates Ser139 in the histone H2AX variant (uH2AX); which is a well-known marker of DNA double-strand breaks (An et al., 2010).

Double-strand breaks can occur endogenously due to the production of reactive oxygen species during metabolism or due to V(D)J recombination during differentiation in the immune system, and exogenously due to ionizing radiation, radiomimetic drugs such as bleomycin, and inhibitors topoisomerase II, such as etoposide and doxorubicin. Therefore, DNA-PK inhibitors are likely to increase the damaging effects of such agents. DNA-PK inhibitors may also be effective as monotherapy in tumors with high endogenous levels of DNA damage caused by defects in other DNA repair pathways, such as HR and mismatch repair. For example, DNA-PK inhibitors have been shown to be effective as monotherapy against ATM-defective lymphoma types (Riabinska et al., 2013). ATM is important for repair through HR, and if cancer cells are deficient in ATM, then these cells are dependent on NHEJ for their survival. Synthetic lethal-type interactions have also been demonstrated for DNA-PK and MSH3 (Deitlein et al., 2014). DNA-PK is a member of the phosphatidylinositol 3-kinase (PIKK) family of protein kinases and is a disadvantage of earlier generation DNA-PK inhibitors such as NU7026, NU7441,

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KU-0060648 and CC-115 have unsatisfactory selectivity against other members of the PIKK family. However, these compounds demonstrated therapeutic potential for targeting DNA-PK, which is consistent with the known mechanisms of action of the DNA-PK protein. For example, NU7026 and KU-0060648 can enhance the cytotoxicity of topoisomerase II inhibitors (Willmore et al, 2004; Munck et al., 2012), and NU7441 enhanced the effect of ionizing radiation in breast cancer models (Ciszewski et al., 2014).

Accordingly, there is a need for DNA-PK inhibitors that are selective, exhibit good bioavailability, and are suitable for dosage administration. In addition, there is a need for DNA-PK inhibitors with a longer half-life to provide increased duration of action with chronic dosing.

Brief description of the invention

Briefly, the present specification specifically describes a compound of formula (I)

(O, or a pharmaceutically acceptable salt thereof, wherein A ¹ is N or CR ^2A , A ² is N or CR ^2B , and A ³ is N or CR ^2C , wherein not more than one of A ¹ , A ^2, and A ³ represents N;

R ¹ represents C _4-6 cycloalkyl or 4-6 membered heterocycloalkyl containing one heteroatom selected from O, S and N, wherein C _4-6 cycloalkyl or 4-6 membered heterocycloalkyl is optionally substituted by one or more groups selected from fluorine, C _1-3 alkyl (optionally substituted by a group selected from hydroxyl and C _1-2 alkoxy), cyclopropyl, hydroxyl, NH ₂ , dioxo, C(O)C _1-2 alkyl, azetidinyl and oxetanyl;

R ^2A , R ^2B and R ^2C are each independently hydrogen, methyl or methoxy.

The present description in particular also describes a pharmaceutical composition which contains a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient.

The present specification also specifically describes a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in therapy.

The present specification also specifically describes a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of cancer.

The present specification also specifically describes a compound of formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of cancer.

Also specifically described herein is a method of treating cancer in a warm-blooded animal in need of such treatment, which comprises administering to said warm-blooded animal a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

Brief description of graphic materials

In fig. Figure 1 shows XRPD results for Form A of 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin-8-one ( compound A, example 44).

In fig. Figure 2 shows the DSC results for Form A of 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin-8-one ( compound A, example 44).

In fig. Figure 3 shows XRPD results for form A^)-7-methyl-2-((7-methylcinnolin-6-yl)amino)9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purine-8- she (compound B, example 21).

In fig. Figure 4 shows the DSC results for form A^)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purine-8- she (compound B, example 21).

In fig. Figure 5 shows XRPD results for Form A 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H -purin-8-one (compound C, example 52).

Description of Illustrative Embodiments

Many embodiments of the present invention have been described in detail throughout the present specification and will be apparent to the reader skilled in the art. The present invention should not be interpreted as being limited to any particular embodiment(s).

In the first embodiment, a compound of formula (I) is provided

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or a pharmaceutically acceptable salt thereof, wherein A ¹ is N or CR ^2A , A ² is N or CR ^2B and A ³ is N or CR ^2C , wherein at most one of A ¹ , A ² and A ³ is N ;

R ¹ represents C _4-6 cycloalkyl or 4-6 membered heterocycloalkyl containing one heteroatom selected from O, S and N, wherein C _4-6 cycloalkyl or 4-6 membered heterocycloalkyl is optionally substituted by one or more groups selected from fluorine, C _1-3 alkyl (optionally substituted by a group selected from hydroxyl and C _1-2 alkoxy), cyclopropyl, hydroxyl, NH ₂ , dioxo, C(O)C _1-2 alkyl, azetidinyl and oxetanyl;

R ^2A , R ^2b and R ^2C are each independently hydrogen, methyl or methoxy.

C4-6cycloalkyl is a saturated non-aromatic carbocyclic ring containing no heteroatoms. C _4-6 cycloalkyl is any such carbocyclic ring containing from 4 to 6 carbon atoms. C _4-6 cycloalkyl groups include cyclobutyl, cyclopentyl and cyclohexanyl, for example cyclohexanyl.

The term cyclohexanyl refers to a carbocyclic ring containing six carbon atoms. 1-Hydroxycyclohex-4-yl groups and 4-hydroxycyclohex-1-yl groups have the same structure, which is shown below.

BUT.

The cis-1-hydroxy-cyclohex-4-yl group is equivalent to cis-4-hydroxy-cyclohex-1-yl and has the following structure:

BUT^/x.

In the structures presented above, the dotted line indicates the position at which the corresponding group is bound.

A 4-6 membered heterocycloalkyl is a saturated non-aromatic ring containing one heteroatom independently selected from nitrogen, oxygen or sulfur, the remaining ring members being carbon. 4-6 membered heterocycloalkyl groups include piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, azetidinyl and oxetanyl, for example piperidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, oxetanyl and pyrrolidinyl. To avoid ambiguity, substituents on the heterocycloalkyl ring can be linked either through a carbon atom or through a heteroatom.

The term dioxo means two oxo substituents that are attached to the same atom. Examples of dioxo substitution include those where R ¹ is thianyl, which may also be referred to as tetrahydrothiopyranyl, wherein the ring sulfur atom is replaced by two oxo groups, i.e. tetrahydrothiopyran-1,1-dioxide.

The prefix C _pq in C _p-q alkyl and other terms (where p and q are integers) indicates the range of the number of carbon atoms present in the group and, unless otherwise noted, the alkyl groups and alkoxy groups containing the required number of carbon atoms , can be branched or unbranched. C _1-3 alkyl groups include methyl (Me), ethyl (Et), n-propyl and isopropyl, for example methyl and ethyl.

The term C _p-q alkoxy includes -O-C _p-q alkyl groups. C _1-2 alkoxy groups include methoxy and ethoxy, for example methoxy.

The use of the term does not necessarily imply that the subsequent feature may or may not exist. In this regard, the use of the term does not necessarily include cases when this characteristic is present, as well as cases when this characteristic is absent. For example, a group optionally substituted with one methoxy group includes groups with and without a methoxy substituent.

The term substituted means that one or more hydrogen atoms (for example, 1 or 2 hydrogen atoms, or alternatively 1 hydrogen atom) in the designated group are replaced.

- 3 043669 by any substituent(s) (e.g. 1 or 2 substituents or alternatively 1 substituent), provided that any atom(s) bearing the substituent retain(s) the permissible valence. Combinations of substituents cover only stable compounds and stable synthetic intermediates. The term stable means that the corresponding compound or intermediate is sufficiently stable to be isolated and has utility either as a synthetic intermediate or as a drug having potential therapeutic utility. If a group is not described as substituted or optionally substituted, it should be considered unsubstituted (i.e., none of the hydrogen atoms in the designated group have been replaced).

The term pharmaceutically acceptable is used to indicate that an entity (eg, a salt, dosage form, or excipient) is suitable for use in patients. An illustrative list of pharmaceutically acceptable salts can be found in the Handbook of Pharmaceutical Salts: Properties, Selection and Use, P.H. Stahl and C. G. Wermuth, editors, Weinheim/Zurich:WileyVCH/VHCA, 2002.

In a further embodiment, any of the embodiments defined herein (e.g., the embodiment of claim 1) is provided, provided that one or more specific examples (e.g., one, two, or three specific examples) selected from the group consisting of examples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 , 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 , 50, 51 and 52, are individually excluded.

In one embodiment, R ¹ is cyclohexanyl or a 4-6 membered heterocycloalkyl containing one heteroatom selected from O, N or S.

In another embodiment, R ¹ is a 4-6 membered heterocycloalkyl containing one heteroatom selected from O or N.

In another embodiment, R ¹ is a 4-6 membered heterocycloalkyl containing one N heteroatom.

In another embodiment, R ¹ is selected from cyclohexanyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl and tetrahydrothiopyranyl.

In another embodiment, R ¹ is selected from pyrrolidinyl and piperidinyl.

In another embodiment, R ¹ is selected from cyclohexanyl, oxetan-3-yl, tetrahydrofuran-3-yl, tetrahydropyran-3-yl, tetrahydropyran-4-yl, pyrrolidin-3-yl, piperidin-4-yl and tetrahydrothiopyran-4-yl.

In another embodiment, R ¹ is selected from pyrrolidin-3-yl and piperidin-4-yl.

In one embodiment, R ¹ is optionally substituted with one or two substituents selected from fluorine, methyl, ethyl, hydroxyl, NH ₂ , dioxo, C(O)Me and oxetanyl, where ethyl is optionally substituted with hydroxyl or methoxy. In one embodiment, R ¹ is optionally substituted with fluorine, methyl, ethyl, hydroxyl, NH2 and oxetanyl. In one embodiment, R ¹ is optionally substituted with fluorine or methyl.

In one embodiment, R ¹ is pyrrolidinyl or piperidinyl and is optionally substituted with one or two substituents selected from fluorine, methyl, ethyl, hydroxyl, NH ₂ and oxetanyl.

In one embodiment, R ¹ is cyclohexanyl, optionally substituted with hydroxyl, methyl or NH2.

In another embodiment, R ¹ is oxetan-3-yl.

In another embodiment, R ¹ is tetrahydrofuran-3-yl.

In another embodiment, R ¹ is tetrahydropyran-3-yl or tetrahydropyran-4-yl.

In another embodiment, R ¹ is pyrrolidin-3-yl, optionally substituted with methyl. In another embodiment, R ¹ is pyrrolidin-3-yl, optionally substituted with fluorine.

In another embodiment, R ¹ is 4-fluoropyrrolidin-3-yl.

In another embodiment, R ¹ is piperidin-4-yl, optionally substituted with a group selected from methyl, ethyl (unsubstituted or substituted with methoxy or hydroxyl), C(O)Me and oxetan-3-yl. In another embodiment, R ¹ is piperidin-4-yl, optionally substituted with methyl.

In another embodiment, R ¹ is 1-methylpiperidin-4-yl.

In another embodiment, R ¹ is dioxidotetrahydro-2H-thiopyran-4-yl.

In one embodiment, A ¹ is CR ^2A , A ² is CR ^2B , and A ³ is CR ^2C

In another embodiment, A ¹ is N, A ² is CR ^2B , and A ³ is CR ^2C .

In another embodiment, A ² is N, A ¹ is CR ^2A and A ³ is CR ^2C .

In another embodiment, A ³ is N, A ¹ is CR ^2A , and A ² is CR ^2C .

In another embodiment, A ¹ is CR ^2A or N, A ² is CR ^2B , and

And ³ represents CR ^2C .

In one embodiment, R ^2A is hydrogen. In one embodiment, R ^2B is hydrogen. In one embodiment, R ^2C is hydrogen.

In another embodiment, one, two or three groups selected from ^R2A , ^R2B and ^R2C are independently selected from methyl and methoxy, and any remaining ^R2A , ^R2B and/or ^R2C groups are hydrogen.

In another embodiment, one, two or three groups selected from R ^2A , R ^2B and R ^2C are methyl.

In another embodiment, one, two or three groups selected from R ^2A , R ^2B and R ^2C are methoxy.

In another embodiment, one or two groups selected from R ^2A , R ^2B and R ^2C are independently selected from methyl and methoxy.

In another embodiment, one or two groups selected from R ^2A , R ^2B and R ^2C are methyl.

In another embodiment, one or two groups selected from R ^2A , R ^2B and R ^2C are methoxy.

In another embodiment, one group selected from R ^2A , R ^2B and R ^2C is methyl.

In another embodiment, one group selected from R ^2A , R ^2B and R ^2C is methoxy.

The present invention also provides compounds of formula (I) wherein A ¹ is N or CR ^2A , A ² is N or CR ^2B and A ³ is N or CR ^2C , wherein at most one of A ¹ , A ² and A ³ represents N;

R ¹ is cyclohexanyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl or tetrahydrothiopyranyl and is optionally substituted with one or two groups selected from fluorine, methyl, ethyl, hydroxyl, NH ₂ , dioxo, C(O)Me and oxetanyl, where ethyl optionally substituted with hydroxyl or methoxy; and R ^2A , R ^2B and R ^2C are each independently hydrogen, methyl or methoxy.

The present invention also provides compounds of formula (I) wherein A ¹ is N or CR ^2A , A ² is N or CR ^2B and A ³ is N or CR ^2C , wherein at most one of A ¹ , A ² and A ³ represents N;

R ¹ is pyrrolidinyl or piperidinyl and is optionally substituted with one or two groups selected from fluorine, methyl, ethyl, hydroxyl, NH2, dioxo and oxetanyl, where ethyl is optionally substituted with hydroxyl or methoxy; and R ^2A , R ^2B and R ^2C are each independently hydrogen, methyl or methoxy.

In one embodiment

A ¹ is N or CR ^2A , A ² is CR ^2B and A ³ is CR ^2C ;

R ¹ is cyclohexanyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl or tetrahydrothiopyranyl and is optionally substituted with one or two groups selected from fluorine, methyl, ethyl, hydroxyl, NH2, dioxo, C(O)Me and oxetanyl, where ethyl is optional substituted with hydroxyl or methoxy; and R ^2A , R ^2B and R ^2C are each independently hydrogen, methyl or methoxy.

In one embodiment

A ¹ is CR ^2A or N, A ² is CR ^2B and A ³ is CR ^2C ;

R ¹ is pyrrolidinyl or piperidinyl and is optionally substituted with one or two groups selected from fluorine, methyl, ethyl, hydroxyl, NH ₂ , dioxo and oxetanyl, where ethyl is optionally substituted with hydroxyl or methoxy; and R ^2A , R ^2B and R ^2C are each independently hydrogen, methyl or methoxy.

In one embodiment

A ¹ is CR ^2A , A ² is CR ^2B and A ³ is CR ^2C ;

R ¹ is cyclohexanyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl or tetrahydrothiopyranyl and is optionally substituted with one or two groups selected from fluorine, methyl, ethyl, hydroxyl, NH2, dioxo, C(O)Me and oxetanyl, where ethyl is optional substituted with hydroxyl or methoxy; and R ^2A , R ^2B and R ^2C are each independently hydrogen, methyl or methoxy.

In one embodiment

A ¹ is CR ^2A , A ² is CR ^2B and A ³ is CR ^2C ;

R ¹ is pyrrolidinyl or piperidinyl and is optionally substituted with one or two

- 5 043669 groups selected from fluorine, methyl, ethyl, hydroxyl, NH2, dioxo and oxetanyl, where ethyl is optionally substituted with hydroxyl or methoxy; and R ^2A , R ^2B and R ^2C are each independently hydrogen, methyl or methoxy.

In one embodiment, both A ¹ and A ² are CH and A ³ is CR ^2C ;

R ¹ is cyclohexanyl, oxetanyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl or tetrahydrothiopyranyl and is optionally substituted with one or two groups selected from fluorine, methyl, hydroxyl, NH2, dioxo, C(O)Me and oxetanyl; And

R ^2C represents hydrogen, methyl or methoxy.

In another embodiment

A ¹ represents N, and both A ² and A ³ represent CH;

R ¹ is cyclohexanyl, tetrahydrofuranyl, tetrahydropyranyl or piperidinyl and is optionally substituted with hydroxyl, methyl or C(O)Me.

In one embodiment

A ¹ represents CH or N, and both A ² and A ³ represent CH; And

R ¹ represents a 5- or 6-membered heterocycloalkyl containing one heteroatom selected from N or O, optionally substituted with fluorine or methyl.

In another embodiment, A ¹ , A ² and A ³ are each CH; And

R ¹ represents piperidinyl substituted with methyl.

In another embodiment, A ¹ , A ² and A ³ are each CH; And

R ¹ represents pyrrolidinyl substituted with fluorine.

In another embodiment

A ¹ represents N, and A ² and A ³ represent CH; And

R ¹ represents tetrahydrofuranyl.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein the compound is selected from the group consisting of

9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinazolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

9-(1-acetylpiperidin-4-yl)-2-((2,7-dimethylquinoxalin-6-yl)amino)-7-methyl7,9-dihydro-8R-purin-8-one;

9-(1-acetylpiperidin-4-yl)-2-((3,7-dimethylquinoxalin-6-yl)amino)-7-methyl7,9-dihydro-8R-purin-8-one;

9-((1g,4g)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

9-((1g,4g)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)7,9-dihydro-8R-purin-8-one;

2-((4,7-dimethylquinolin-6-yl)amino)-9-((1 g,4g)-4-hydroxycyclohexyl)-7methyl-7,9-dihydro-8R-purin-8-one;

9-((1g,4g)-4-hydroxycyclohexyl)-2-((4-methoxy-7-methylquinolin-6-yl)amino)7-methyl-7,9-dihydro-8R-purin-8-one;

9-((1g,4g)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)7,9-dihydro-8R-purin-8-one;

9-((1y45)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

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9-((15,45)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)7,9-dihydro-8R-purin-8-one;

9-((15,45)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)7,9-dihydro-8//-purin-8-one;

9-((15.45)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8//-purin-8-one ;

9-((1g,4g)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8//-purin-8-one ;

9-((15.45)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylcinnolin-6yl)amino)-7,9-dihydro-8//-purin-8-one ;

9-((1g,4g)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylcinnolin-6yl)amino)-7,9-dihydro-8R-purin-8-one;

9-((1g,4g)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6yl)amino)-7,9-dihydro-8//-purin-8-one ;

9-((15.45)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6yl)amino)-7,9-dihydro-8R-purin-8-one;

9-((15.45)-4-hydroxy-1-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6yl)amino)-7,9-dihydro-8//-purin-8-one ;

(E-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9dihydro-8R-purin-8-one;

(E-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9dihydro-8R-purin-8-one;

(7?)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9dihydro-8R-purin-8-one;

(7?)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9dihydro-8R-purin-8-one;

(R)-7-methyl-2-((7-methyl cinol in-6-yl)am and no)-9-(tetrahydro-2R-pyran-3-yl)7,9-dihydro-8R-purine -8-she;

(7?)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2//-pyran-3-yl)7,9-dihydro-8H-purine-8 -she;

($)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydro-2//-pyran-3-yl)7,9-dihydro-8//-purin- 8-she;

($)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2R-pyran-3-yl)7,9-dihydro-8R-purin-8-one;

7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydro-2//-pyran-4-yl)-7,9dihydro-8H-purin-8-one;

7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(tetrahydro-2/-pyran-4-yl)-7,9dihydro-8R-purin-8-one;

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7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2R-pyran-4-yl)-7,9dihydro-8R-purin-8-one;

7-methyl-2-((7-methylquinazolin-6-yl)amino)-9-(tetrahydro-2R-pyran-4-yl)-7,9dihydro-8R-purin-8-one;

2-((2,7-dimethylquinoxalin-6-yl)amino)-7-methyl-9-(tetrahydro-2R-pyran-4-yl)7,9-dihydro-8R-purin-8-one;

2-((3,7-dimethylquinoxalin-6-yl)amino)-7-methyl-9-(tetrahydro-2R-pyran-4-yl)7,9-dihydro-8R-purin-8-one;

9-(1,1-otetrahydro-2R-thiopyran-4-yl)-7-methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8R-purin-8-one;

7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(oxetan-3-yl)-7,9-dihydro-8Japurin-8-one;

7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(piperidin-4-yl)-7,9-dihydro-8Japurin-8-one;

9-((3Q47?)-3-fluoropiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

9-((15.45)-4-amino-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8R-purin-8-one;

9-((1g,4g)-4-amino-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8R-purin-8-one;

(7?)-7-methyl-9-(1-methylpyrrolid in-3-yl)-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

(5)-7-methyl-9-(1-methylpyrrolidin-3-yl)-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(1-methylpiperidin-4-yl)-7,9dihydro-8R-purin-8-one;

7-methyl-9-(1-methyl piperidin-4-yl)-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

7-methyl-9-(1-methylpiperidin-4-yl)-2-((7-methylquinoxalin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

9-((35.47?)-3-fluoro-1-methylpiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8R-purine- 8-she;

7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(1-(oxetan-3-yl)piperidin-4-yl)7,9-dihydro-8R-purin-8-one ;

9-(1-(2-hydroxyethyl)piperide in-4-yl)-7-methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8R-purin-8-one;

9-(1-(2-methoxyethyl)piperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)7,9-dihydro-8R-purin-8-one;

9-(1-ethylpiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8R-purin-8-one;

9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8R-purin-8-one and

9-((37?,47?)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)7,9-dihydro-8R-purine-8- she.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein the compound is selected from the group consisting of

- 8 043669

7-methyl-9-(1-methylpiperidin-4-yl)-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8//-purin-8-one;

0-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9dihydro-8//-purin-8-one and

9-((3D47?)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)7,9-dihydro-8//-purin-8-one .

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein the compound is 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7methylquinolin-6-yl)amino)-7 ,9-dihydro-8H-purin-8-one (also called compound A).

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein the compound is (S)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9(tetrahydrofuran-3-yl )-7,9-dihydro-8H-nurin-8-one (also called compound B).

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided, wherein the compound is 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7methylquinoline-6- yl)amino)-7,9-dihydro-8H-purin-8-one (also called compound C).

The compounds and salts described herein may exist in solvated forms and unsolvated forms. For example, the solvated form may be a hydrated form such as a hemihydrate, monohydrate, dihydrate, trihydrate, or other degree of hydration. The present invention covers all such solvated and unsolvated forms of the compounds of formula (I), in particular in those cases where such forms have DNA-PK inhibitory activity, which is, for example, measured using the tests described herein.

The atoms of the compounds and salts described herein may exist as isotopes thereof. The present invention covers all compounds of formula (I) where an atom is replaced by one or more isotopes thereof (for example, a compound of formula (I) where one or more carbon atoms is an 11C or ^13C carbon isotope, or where one or more hydrogen atoms are is the ^2H or ^3H isotope, or where one or more nitrogen atoms is the ^15N isotope, or where one or more oxygen atoms is the ^17O or ^18O isotope).

Certain compounds and salts described herein include one or more chiral centers (ie, centers of asymmetricity). Included in the present invention is any optically active or racemic form of a compound of formula (I) that has DNA-PK inhibitory activity as measured, for example, by the assays described herein. Where a structure or chemical name herein does not indicate chirality, the structure or name is intended to cover any single stereoisomer (i.e., any single chiral isomer) corresponding to such structure or name, and any mixture of stereoisomers (for example, a racemate). In some embodiments, a single stereoisomer is obtained by isolating it from a mixture of isomers (eg, a racemate) using, for example, chiral chromatographic separation. In other embodiments, a single stereoisomer is obtained by direct synthesis, for example, from a chiral starting material.

In one embodiment, there is provided a compound of formula (I) or a pharmaceutically acceptable salt thereof that is a single enantiomer present in an enantiomeric excess (% ee) of >95, >98, or >99%. Preferably, an individual enantiomer is present in an enantiomeric excess (% ee) of >99%.

In another embodiment, a pharmaceutical composition is provided that contains a compound of formula (I) that is a single enantiomer present in an enantiomeric excess (% ee) of >95, >98, or >99%, or a pharmaceutically acceptable salt thereof , in combination with one or more pharmaceutically acceptable excipients. Preferably, an individual enantiomer is present in an enantiomeric excess (% ee) of >99%.

Certain compounds of formula (I) and pharmaceutically acceptable salts of any such compounds exist as diastereomers.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided in a diastereomeric excess (% d.i.) of >95, >98 or >99%. In one embodiment, the compound of formula (I) or a pharmaceutically acceptable salt thereof is present in a diastereomeric excess (% d.i.) of >99%.

Some of the compounds of formula (I) may be crystalline and may have more than one crystalline form. It should be understood that the present invention covers any crystalline or amorphous form or mixtures thereof that have properties useful for DNA-PK inhibitory activity. It is well known how to determine the effectiveness of a crystalline or amorphous form using standard tests described later in this document.

It is well known that crystalline materials can be analyzed using conventional techniques such as, for example, X-ray powder diffraction (hereinafter XRPD) analysis and differential scanning calorimetry (hereinafter DSC).

As an example, the compound of Example 44, 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7methylquinolin-6-yl)amino)-7,9-dihydro-8H-purine-8- It is characterized by crystallinity, and one crystalline form, Form A, has been identified.

Accordingly, in an additional aspect there is provided Form A of Compound A (Example 44, 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro- 8H-purin-8-one).

The present invention provides a crystalline form, Form A of Compound A, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of approximately 7.1° as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound A, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of approximately 8.5° as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound A, which is characterized by an XRPD pattern with at least two characteristic peaks at 2-theta angle values of approximately 7.1 and 8.5° as measured using CuKa radiation .

The present invention provides a crystalline form, Form A of Compound A, which is characterized by an XRPD pattern with characteristic peaks at 2-theta angle values of approximately 7.1, 8.5, 12.7, 14.2, 15.4, 16.3, 18.8, 19.8, 21.5, 26.2°, as measured using CuKa radiation.

In accordance with the present invention, there is provided a crystalline form, Form A of Compound A, which is characterized by an XRPD XRD pattern substantially the same as the XRD pattern shown in FIG. 1.

The present invention provides a crystalline form, Form A of Compound A, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of 7.1° plus or minus 0.2° 2-theta as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound A, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle value of 8.5° plus or minus 0.2° 2-theta as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound A, which is characterized by an XRPD with at least two characteristic peaks at a 2-theta angle of 7.1 and 8.5°, which values may vary by plus or minus minus 0.2° 2-theta angle as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound A, which is characterized by an XRPD X-ray diffraction pattern with characteristic peaks at 2-theta angle values of 7.1, 8.5, 12.7, 14.2, 15.4, 16 ,3, 18.8, 19.8, 21.5, 26.2°, where the indicated values may vary by plus or minus 0.2° 2-theta, as measured using CuKa radiation.

The results of DSC analysis of Form A of Compound A demonstrated a melting endotherm with an onset at approximately 245°C and a peak at approximately 246°C (FIG. 2).

In an additional aspect, Form A of Compound B is provided (Example 21, (S)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9 -dihydro-8H-purin-8-one).

The present invention provides a crystalline form, Form A, of Compound B that is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of approximately 9.7° as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound B, which is characterized by an XRPD pattern with at least one characteristic peak at a 2-theta angle of approximately 12.9°.

The present invention provides a crystalline form, Form A of Compound B, which is characterized by XRPD with at least two characteristic peaks at 2-theta angle values of approximately 9.7 and 12.9° as measured using CuKa radiation.

- 10 043669

The present invention provides a crystalline form, Form A of Compound B, which is characterized by an XRPD pattern with characteristic peaks at 2-theta angle values of approximately 9.7, 12.5, 12.9, 15.8, 17.6, 17.9, 19.4, 21.0, 26.0, 26.4°, as measured using CuKa radiation.

In accordance with the present invention, there is provided a crystalline form, Form A, of Compound B that exhibits an XRPD pattern substantially the same as the XRPD pattern shown in FIG. 3.

The present invention provides a crystalline form, Form A of Compound B, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of 9.7° plus or minus 0.2° 2-theta as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound B, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of 12.9° plus or minus 0.2° 2-theta as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound B, which is characterized by an XRPD pattern with at least two characteristic peaks at 2-theta angle values of 9.7 and 12.9°, where these values may deviate by plus or minus minus 0.2° 2-theta, as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound B, which is characterized by an XRPD X-ray diffraction pattern with characteristic peaks at 2-theta angle values of 9.7, 12.5, 12.9, 15.8, 17.6, 17 ,9, 19.4, 21.0, 26.0, 26.4°, where the indicated values may vary by plus or minus 0.2° 2-theta, as measured using CuKa radiation.

The results of DSC analysis of Compound B Form A showed a melting endotherm with an onset at approximately 227°C and a peak at approximately 228°C (FIG. 4).

According to a further aspect, Form A of Compound C is provided (Example 52, 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino) -7,9-dihydro-8H-purin-8-one).

The present invention provides a crystalline form, Form A of Compound C, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of approximately 7.3° as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound C, which is characterized by an XRPD pattern with at least one characteristic peak at a 2-theta angle of approximately 15.0°.

The present invention provides a crystalline form, Form A of Compound C, which is characterized by XRPD with at least two characteristic peaks at 2-theta angle values of approximately 7.3 and 15.0° as measured using CuKa radiation.

The present invention provides a crystalline form, Form A, of Compound C, which is characterized by an XRPD pattern with characteristic peaks at 2-theta angle values of approximately 7.3, 15.0, 14.6, 26.5, 12.2, 26.0, 17.0, 15.9, 27.3, 10.8°, as measured using CuKa radiation.

In accordance with the present invention, there is provided a crystalline form, Form A of Compound C, which exhibits an XRPD pattern substantially the same as the XRPD pattern shown in FIG. 5.

The present invention provides a crystalline form, Form A of Compound C, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of 7.3° plus or minus 0.2° 2-theta as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound C, which is characterized by an XRPD X-ray diffraction pattern with at least one characteristic peak at a 2-theta angle of 15.0° plus or minus 0.2° 2-theta as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound C, which is characterized by an XRPD pattern with at least two characteristic peaks at 2-theta angle values of 7.3 and 15.0°, where these values may deviate by plus or minus minus 0.2° 2-theta, as measured using CuKa radiation.

The present invention provides a crystalline form, Form A of Compound C, which is characterized by an XRPD pattern with characteristic peaks at 2-theta angle values of 7.3, 15.0, 14.6, 26.5, 12.2, 26 ,0, 17.0, 15.9, 27.3, 10.8°, where the indicated values

- 11 043669 may deviate by plus or minus 0.2° 2-theta, as measured using CuKa radiation.

If the present invention is said to provide a crystalline form that is Form A of Compound A, Form A of Compound B, and Form A of Compound C, the degree of crystallinity is preferably greater than about 60%, more preferably greater than about 80%, preferably greater than about 90% or greater preferably greater than about 95%. Most preferably, the degree of crystallinity is greater than about 98%.

It will be appreciated that the 2-theta angle values in an XRPD radiograph may vary slightly from one device to another or from one sample to another, and therefore the values given should not be considered absolute.

It is known that it is possible to obtain an XRPD radiograph that is characterized by one or more measurement errors depending on the measurement conditions (such as the equipment or device used). In particular, it is common knowledge that the intensity values in an XRPD pattern can fluctuate depending on the measurement conditions. Therefore, it should be understood that Form A of Compound A, Form A of Compound B, and Form A of Compound C of the present invention are not limited to crystals that provide XRPD patterns identical to the XRPD patterns shown in FIG. 1, 3 and 5, and any crystals providing XRPD patterns are substantially the same as those shown in FIGS. 1, 3 and 5 are within the scope of the present invention. One skilled in the art of XRPD can conclude that the XRPD radiographs are substantially identical.

Those skilled in the art of XRPD will recognize that the relative intensity of the peaks can be influenced, for example, by grains larger than 30 microns and aspect ratios far from unity, which can affect sample analysis. One skilled in the art will also appreciate that the position of the reflections can be influenced by the precise height at which the sample is placed in the diffractometer and the calibration of the diffractometer zero. The flatness of the sample surface may also have a minor effect. Therefore, the XRD data presented should not be taken as absolute values. (Jenkins, R. & Snyder, R.L., 'Introduction to X-Ray Powder Diffractometry', John Wiley & Sons 1996; Bunn, C.W. (1948), Chemical Crystallography, Clarendon Press, London; Klug, H.P. & Alexander, L.E. (1974 ), X-Ray Diffraction Procedures).

Typically, the measurement error in diffraction angle on a powder X-ray diffraction pattern is approximately plus or minus 0.2° 2-theta, and this level of measurement error must be taken into account when considering the XRPD patterns in FIG. 1, 3 and 5 and when reading the table. A, B, and C. Additionally, it should be understood that intensities may fluctuate depending on experimental conditions and sample preparation (preferred orientation).

Compounds of formula (I) can be prepared, for example, by reacting a compound of formula (II)

or a salt thereof, wherein R ¹ is as defined in any of the embodiments herein, or a protected form thereof, and X represents a leaving group (e.g., a halogen atom, such as a chlorine atom), with a compound of formula (III)

or a salt thereof, wherein A ¹ , A ² and A ³ are as defined in any of the embodiments herein. Advantageously, the reaction is carried out in a suitable solvent (eg 1,4-dioxane) in the presence of a base (eg cesium carbonate) and optionally in the presence of a suitable catalyst (eg Brettphos, 3rd generation catalyst) at a suitable temperature (eg temperature in the range of approximately 80 -100°C).

Therefore, the compounds of formula (II) or (III) and their salts are useful as intermediates in the preparation of compounds of formula (I) and provide an additional option

- 12 043669 distribution. In one embodiment, there is provided a compound of formula (II) or a salt thereof, wherein R ¹ is C _4-6 cycloalkyl or a 4-6 membered heterocycloalkyl containing one heteroatom selected from O, S and N, wherein C _4-6 cycloalkyl or 4-6 membered heterocycloalkyl is optionally substituted with one or more groups selected from fluoro, C _1-3 alkylα (optionally substituted by a group selected from hydroxyl and C _1-2 alkoxy), cyclopropyl, hydroxyl, NH2, dioxo, C(O )C _1-2 alkyl, azetidinyl and oxetanyl; And

X represents a leaving group.

In one embodiment, X represents a halogen atom or a triflate group. In one embodiment, X is a chlorine atom.

In any of the embodiments in which a compound of formula (II) or (III) or a salt thereof is mentioned, it should be understood that such salts need not be pharmaceutically acceptable salts.

Compounds of formula (II) can be prepared, for example, by reacting a compound of formula (IV)

wherein R ¹ is as defined in any of the embodiments herein, and X represents a leaving group (eg, an iodine, bromine or chlorine atom or a triflate group) with a methylating agent. Suitable methylating agents include methyl iodide, DMF-DMA.

Compounds of formula (IV) can be prepared, for example, by reacting a compound of formula (V)

where R ¹ is as defined in any of the embodiments herein;

R ^a represents hydrogen; And

X represents a leaving group (eg iodine, bromine, chlorine or triflate group) with diphenylphosphoryl azide (DPPA).

The reaction can be carried out under standard conditions well known to those skilled in the art, for example, using DPPA, triethylamine, THF, reflux.

Therefore, the compounds of formulas (IV) and (V) are useful as intermediates in the preparation of compounds of formula (I) and provide a further embodiment.

The compounds of formulas (IV) and (V) can be prepared by methods similar to those shown in the Examples section.

The compound of formula (III) can be prepared, for example, by reacting the compound of formula (VI) o ₂ n

A ¹ il N (VI), where A ¹ , A ² and A ³ are as defined in any of the embodiments herein, with a reducing agent. Suitable reducing agents include 10% Pd/C and hydrogen, 10% Pd/C and ammonium formate, iron/ammonium chloride.

The compound of formula (III) can also be prepared, for example, by reacting the compound of formula (VII)

- 13 043669

(VII), wherein A ¹ , A ² and A ³ are as defined in any of the embodiments herein; And

X represents a leaving group (eg a bromine or chlorine atom or a triflate group).

Advantageously, the reaction is carried out in a suitable solvent (eg 1,4-dioxane) in the presence of a base (eg sodium t-butoxide) and an equivalent amine (eg benzophenonymine) and optionally in the presence of a suitable catalyst (eg tris(dibenzylideneacetone)dipalladium) and ligand (BINAP) at a suitable temperature (eg, a temperature in the range of approximately 80-100°C).

The compound of formula (III) can also be prepared, for example, by reacting the compound of formula (VIII)

2 ^(VIII) 'where A ¹ , A ² and A ³ are as defined in any of the embodiments herein; a

PG is a suitable protecting group, for example BOC or benzophenonymine.

Advantageously, the reaction is carried out in a suitable solvent (eg methanol) in the presence of an acid (hydrochloric acid) at a suitable temperature (eg temperature in the range of about 20°C).

The compounds of formulas (III), (VI), (VII) and (VIII) can be prepared by methods similar to those shown in the Examples section.

It will be understood that certain substituents from the various ring substituents in the compounds of the present invention can be introduced through standard aromatic ring substitution reactions or obtained through conventional functional group modifications either before or immediately after the methods mentioned above, and are therefore included in method aspect of the present invention. For example, compounds of formula (I) can be converted into additional compounds of formula (I) through standard aromatic ring substitution reactions or through conventional functional group modifications. Such reactions and modifications include, for example, the introduction of a substituent through a substitution reaction on the aromatic ring, the reduction of substituents, the alkylation of substituents, and the oxidation of substituents. The reagents and reaction conditions for such procedures are well known in the field of chemistry. Specific examples of aromatic ring substitution reactions include introduction of a nitro group using concentrated nitric acid, introduction of an acyl group using, for example, an acyl halide and a Lewis acid (such as aluminum trichloride) under Friedel-Crafts conditions; introducing an alkyl group using an alkyl halide and a Lewis acid (such as aluminum trichloride) under Friedel Crafts conditions and introducing a group representing a halogen. Specific examples of modifications include reduction of the nitro group to an amino group, for example, by catalytic hydrogenation with a nickel catalyst or treatment with iron in the presence of hydrochloric acid with heating; oxidation of alkylthio to alkylsulfinyl or alkylsulfonyl.

It will also be understood that in some of the reactions mentioned herein, it may be necessary/desirable to protect any sensitive groups in the compounds. Cases where protection is necessary or desirable and suitable methods of protection are known to those skilled in the art. Traditional protecting groups can be used according to standard practice (for illustration, see T. W. Green, Protective Groups in Organic Synthesis, John Wiley and Sons, 1991). Thus, if the reactants include groups such as amino, carboxy or hydroxy, in some of the reactions mentioned herein, protection of this group may be desirable.

The compounds of formulas (I), (II) and (III) and any intermediates used for their preparation can be prepared by methods similar to those shown in the Examples section.

Biological analyses.

The following assays were performed to measure the effects of the compounds described herein:

a) analysis of activity against the DNAPK enzyme;

b) DNAPK activity assay in cells.

When describing the analyzes as a whole, the following is implied.

i) The following abbreviations have been used:

DMSO=dimethyl sulfoxide;

DTT=dithiothreitol;

EDTA=ethylenediaminetetraacetic acid;

TR-FRET=time resolved fluorescence inductive resonance energy transfer;

ATP=adenosine triphosphate;

DTT=dithiothreitol;

DNA=deoxyribonucleic acid;

HEPES=(2-hydroxyethyl)-1-piperazineethanesulfonic acid.

ii) The IC ₅₀ value represented the concentration of the test compound that provided 50% inhibition of biological activity.

Analysis.

a) Analysis of activity against the DNAPK enzyme (DNA-PK enzyme).

The DNAPK inhibitory activity of the compounds was determined by TR-FRET, which measures the conversion of a fluorescently tagged peptide substrate to a phosphorylated product. Fluorescently labeled peptide substrate was purchased from Thermo Fisher Scientific. 12-point compound concentration-response curves on a semi-log scale at a maximum concentration of 100 μM were obtained from 10 mM stock solutions of the compound solubilized in DMSO using Echo 555 (Labcyte Inc., Sunnyvale, CA). All assays were performed in Greiner 1536-well white low-well plates (Greiner Bio-One, UK) with a total reaction volume of 3 μL and a final concentration of 1% (v/v) DMSO.

Enzymes and substrates were added separately to compound plates and incubated at room temperature. The kinase reaction was then quenched by adding 3 μl of stopping buffer. After stopping the reaction, the assay plates were read using BMG Pherastar. IC ₅₀ values were calculated using Genedata Screener® software (Genedata, Inc., Basel, Switzerland).

Full-length human DNAPK protein was purified from HeLa cell extract by ion exchange. First, the DNAPK protein was incubated with the compound for 30 min at room temperature in reaction buffer (50 mM Hepes, pH 7.5, 0.01% Brij-35, 10 mM MgCl2 _, 1 mM EGTA, 1 mM DTT, 2 μg/ ml of DNA isolated from calf thymus). The reaction was then initiated by adding ATP and a fluorescently labeled peptide substrate (fluorescein-EPPLSQEAFADLWKK, Thermo Fisher Scientific). The kinase reaction (18 μM ATP, 35 pM DNAPK, 1.6 μM peptide substrate) was quenched after 40 min by adding 3 μl of stopping buffer (20 mM Tris, pH 7.5, 0.02% sodium azide, 0.01% Nonidet -P40, 20 µM EDTA, 4 nM Tb-labeled phospho-p53 antibody [Ser15] The reaction mixture was incubated for an additional hour and the plates were read on a BMG Pherastar.

Data were analyzed and IC ₅₀ values were calculated using Genedata Screener® software (Genedata, Inc., Basel, Switzerland). pIC ₅₀ values were calculated as the negative logarithm of the molar concentration of compound required to produce a 50% reduction in the measured response.

b) Activity assay against DNAPK enzyme in cells (DNA-PK in cells).

Compounds or DMSO (dimethyl sulfoxide) were dispensed from stock plates containing 10 mM compounds in 100% (v/v) DMSO or 100% DMSO directly into cell assay plates using an Echo 555 acoustic dispenser (Labcyte Inc™ ). Stock solutions of compounds at a concentration of 10 mM were diluted 1:100 using an Agilent VPrep liquid pipette with a 96-channel fixed tip head (Agilent Technologies, Santa Clara, CA) to obtain four intermediate dilutions (10 mM, 100, 1 µM, 10 nM). This plate, with intermediate 1:100 dilutions, was then used to measure compounds and DMSO using Echo directly into the cell plates at a range of doses to produce 12 plot points (30, 10, 3.125, 1.25, 0.3, 0 ,1, 0.03125, 0.0125, 0.003, 0.001, 0.0003125, 0.00003 µM) to calculate the _IC50 values of the compounds, with the total DMSO concentration in the assay being 0.3% (v/v). ).

ELISA analysis for DNAPK on cells was performed using the A549 cell line. A549 cells were cultured in cell culture medium consisting of MEM-F12 (F12 Minimal Essential Medium, Sigma No. D6421), 10% (v/v) fetal calf serum and 1% (v/v) 200 mM L -glutamine. After collection, cells were measured into black 384-well Costar plates (#3712, Coining)

- 15 043669 to obtain 15,000 cells per well in a total volume of 40 μl of cell culture medium and incubated overnight at 37°C, 90 relative humidity and 5% CO2 in a rotary incubator. Greiner 781077 all-black 384-well high-binding ELISA plates were coated with 0.5 μg/ml anti-DNAPK antibody (Abcam no. ab1832) in PBS/A overnight at 4°C. The next day, Greiner ELISA plates were washed 3x with PBS-T and blocked with 3% BSA/PBS for ∼2 h before an additional 3x wash with PBS-T.

Test compounds and reference controls were dosed directly into cell plates using a Labcyte Echo 555 acoustic dispenser. Cell plates were then incubated for 1 hour at 37°C prior to treatment with 8 Gy of radiation (XRAD 320, table height 65). Cells were incubated for an additional 1 h before cell culture medium was removed. Lysis buffer (homemade with the addition of protease inhibitor mixture tablets, Roche no. 04693116001, and phosphatase inhibitor tablets, Roche no. 04906837001) was measured at 25 μl/well and the plates were incubated at 4°C for 30 min. Cell lysates (20 μl/well) were transferred to DNAPK antibody-coated ELISA plates using the CyBio Felix liquid dispensing platform, and the ELISA plates were incubated at 4°C overnight.

The next day, ELISA plates were washed 3x with PBS-T and homemade anti-pS2056-DNAPK antibody (0.5 μg/ml in 3% BSA/PBS) was measured at 20 μl/well. The plates were incubated with the antibody for 2 h at room temperature (RT) before washing 3x with PBS-T. HRP-conjugated goat anti-rabbit secondary antibody (1:2000 dilution in 3% BSA/PBS; Cell Signaling #7074) was dispensed at 20 μl/well and plates were incubated at room temperature for 1 h before washing 3 times with PBS-T.

QuantaBlu Substrate Working Solution (Thermo Scientific #15169, prepared according to the manufacturer's instructions) was dispensed at 20 µl/well and the plates were incubated at RT for 1 h before further dispensing 20 µl/well of the QuantaBlu stopping solution provided in the kit (Thermo Scientific #15169). Fluorescence intensity of individual wells was determined using a PerkinElmer EnVision plate reader.

Data were analyzed and IC ₅₀ values were calculated using Genedata Screener® software (Genedata, Inc., Basel, Switzerland). pIC ₅₀ values were calculated as the negative logarithm of the molar concentration of compound required to produce a 50% reduction in the measured response.

c) Enzyme assay with TTC.

The TTC inhibitory activity of the compounds was determined through a Eu LanthaScreen® kinase binding assay performed at ThermoFisher Scientific as part of their biochemical SelectScreen® kinase profiling service. The Eu LanthaScreen® kinase binding assay format involves the binding of an Alexa Fluor® conjugate or tracer to the kinase, which is detected by adding a Eu-labeled antibody to the tag. Binding of the tracer and antibody to the kinase results in high levels of FRET, while displacement of the tracer by a kinase inhibitor results in loss of FRET. The level of FRET measured in the assay is used to determine the binding of the compound.

10-point concentration-response curves for three-fold dilutions of the compound with a maximum concentration of 10 μM were obtained from 10 mM stock solutions of the compound solubilized in DMSO. All assays were performed in Greiner white 384-well low-well plates (cat. no. 784207, Greiner) with a total reaction volume of 16 μL and a final concentration of 1% (v/v) DMSO. 3.84 µl kinase buffer (50 mM HEPES, pH 7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA), 8 µl 2x kinase/antibody mixture (final concentrations 5 nM TTK, 2 nM Eu-labeled anti-GST antibody prepared in kinase buffer) and 4 µl of 4x AlexaFluor®-labeled tracer solution (final concentrations of 30 nM tracer 236 prepared in kinase buffer) were added separately to compound plates, placed on a plate shaker for 30 s, and then incubated for 60 min. at room temperature. The plates were then read using a plate reader for fluorescence imaging. IC ₅₀ values were calculated using XLfit software (IDBS Ltd, Surrey, UK) with curve fitting model number 205 (sigmoidal dose-response curve model).

d) Enzyme assays with Aurora-A, Aurora-B, JAK1, JAK2, JAK3.

The inhibitory activity of the compounds against AURKA, AURKB, JAK1, JAK2 and JAK3 was determined in Z'-LYTE® assays performed by ThermoFisher Scientific as part of their biochemical SelectScreen® kinase profiling service. The Z'-LYTE® biochemical assay format provides a fluorescence-based format with a coupled enzyme and is based on the differential sensitivity of phosphorylated and non-phosphorylated peptides to proteolytic degradation. The peptide substrate is labeled with two fluorophores, one at each end, which are paired for FRET. In the primary reaction, the kinase transfers ATP gamma phosphate to a single tyrosine, serine, or threonine residue in a synthetic FRET peptide. In a secondary reaction, a site-specific protease recognizes and degrades nonphosphorylated FRET peptides. Phosphorylation

- 16 043669

FRET peptides inhibit cleavage by the developing reagent. Cleavage disrupts FRET between donor (i.e., coumarin) and acceptor (i.e., fluorescein) fluorophores on the FRET peptide, while uncleaved phosphorylated FRET peptides maintain FRET. To quantify the reaction progress, the ratio method was used, which calculates the ratio (emission ratio) of donor emission to acceptor emission after excitation of the donor fluorophore at 400 nm. Both cleaved and uncleaved FRET peptides affect fluorescence signals and hence the emission ratio. Based on the emission ratio, the degree of phosphorylation of the FRET peptide can be calculated. The emission ratio will remain low if the FRET peptide is phosphorylated (ie there is no kinase inhibition) and will be high if the FRET peptide is not phosphorylated (ie there is kinase inhibition).

10-point concentration-response curves for three-fold dilutions of the compound with a maximum concentration of 10 μM were obtained from 10 mM stock solutions of the compound solubilized in DMSO. All assays were performed in Corning black 384-well low-volume nonbinding well plates (Cat. No. 4514, Corning) in a total reaction volume of 10 μL and a final concentration of 1% (v/v) DMSO. 2.4 µl kinase buffer (50 mM HEPES, pH7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA), 5 µl 2x peptide/kinase mixture (detailed below for each kinase) and 2.5 µl of 4x ATP solution (prepared in kinase buffer) was added individually to compound plates, placed on a plate shaker for 30 s and then incubated for 60 min. at room temperature. The kinase reaction was then quenched by adding 5 μl of development reagent (proprietary from ThermoFisher Scientific). Assay plates were placed on a plate shaker for 30 s, incubated for 60 min at room temperature, and then read using a fluorescence imaging plate reader. IC ₅₀ values were calculated using XLfit software (IDBS Ltd, Surrey, UK) with curve fitting model No. 205 (sigmoidal dose-response curve model).

Aurora A (AurA): prepared a 2X mixture of AURKA (Aurora A)/Ser/Thr 01 (a ThermoFisher Scientific proprietary product) in 50 mM FLEPES, pH 7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA. The final 10 µl kinase reaction solution was 15 nM AURKA (Aurora A), 2 µM Ser/Thr 01 and 10 µM ATP (each Km) in 50 mM HEPES, pH 7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA. After incubating the kinase reaction for 1 hour, 5 μl of developing reagent was added at a dilution of 1:4096.

Aurora B (AurB): a 2X mixture of AURKB (Aurora B)/Ser/Thr 01 (a ThermoFisher Scientific patented product) was prepared in 50 mM HEPES, pH 7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA. The final 10 µl kinase reaction solution was 23 nM AURKB (Aurora B), 2 µM Ser/Thr 01 and 75 µM ATP (each Km measured for ATP is 81 µM) in 50 mM HEPES, pH 7.5.0 .01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA. After incubating the kinase reaction for 1 hour, 5 μl of developing reagent was added at a dilution of 1:4096.

JAK1: Prepare a 2X mixture of JAK1/Tyr 06 (proprietary of ThermoFisher Scientific) in 50 mM HEPES, pH 6.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA, 0.02% NaN3. The final 10 µl kinase reaction solution was 74 nM JAK1, 2 µM Tyr 06 and 75 µM ATP (each Km measured for ATP is 87 µM) in 50 mM HEPES, pH7.0, 0.01% BRIJ-35, 10 mM MgCb, 1 mM EGTA, 0.01% NaN ₃ . After incubating the kinase reaction for 1 hour, 5 μl of developing reagent was added at a dilution of 1:128.

JAK2: Prepare a 2X mixture of JAK2/Tyr 06 (proprietary of ThermoFisher Scientific) in 50 mM HEPES, pH 7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA. The final 10 µl kinase reaction solution was 0.27 nM JAK2, 2 µM Tyr 06 and 25 µM ATP (each Km measured for ATP is 31 µM) in 50 mM HEPES, pH7.5, 0.01% BRTJ- 35, 10 mM MgCl ₂ , 1 mM EGTA. After incubating the kinase reaction for 1 hour, 5 μl of developing reagent was added at a dilution of 1:128.

JAK3: Prepare a 2X mixture of JAK3/Tyr 06 (proprietary of ThermoFisher Scientific) in 50 mM HEPES, pH 7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ , 1 mM EGTA. The final 10 µL kinase reaction solution consisted of 2.4 nM JAK3, 2 µM Tyr 06 and 10 µM ATP (each Km measured for ATP is 14 µM) in 50 mM HEPES pH 7.5, 0.01% BRTJ-35, 10 mM MgCl ₂ , 1 mM EGTA. After incubating the kinase reaction for 1 hour, 5 μl of developing reagent was added at a dilution of 1:128.

Examples were tested in the above assays and the following data were observed (data shown are the arithmetic average of pIC ₅₀ values observed in two or more experiments).

- 17 043669

Example p!C50 FOR DNAPK enzyme p!C50 FOR DNARK in cells ah p!C50 FOR TTK enzyme p!C50 FOR JAK1 enzyme p!C50 FOR JAK2 enzyme p!C50 FOR JAK3 enzyme p!C50 FOR enzyme AngA p!C50 FOR the enzyme AngB 1 10.0 6.7 5.9 <5 <5.2 <5 <5 <5 2 9.8 7.3 5.8 <5 <5 <5 <5 <5 3 >10 7.2 5.4 <5 <5 <5 <5 <5 4 9.0 6.6 6.0 <5 <5 <5 <5 <5 5 9.1 7.0 5.3 <5 <5 <5 <5 <5 6 >10 8.3 7.2 <5 <5 <5 <5 <5 7 >10 7.3 6.9 <5 <5 <5 <5 <5 8 >10 8.0 6.2 <5 <5 <5 <5 <5 9 >10 8.1 6.1 <5 <5 <5 <5.1 <5 10 >10 8.0 6.4 <5 <5 <5 <5 <5 eleven 9.8 7.7 6.4 <5 <5 <5 <5 <5 12 >10 7.7 6.3 <5 <5 <5 <5 <5 13 9.7 7.8 6.5 <5 <5 <5 <5 <5 14 >10 8.0 7.0 <5 <5 <5 <5 <5 15 >10 7.8 6.6 <5 <5 <5 <5 <5 16 >10 7.8 6.7 <5 <5 <5 <5 <5 17 >10 7.0 6.5 <5 <5 <5 <5 <5 18 9.6 7.3 5.8 <5 <5 <5 <5 <5 19 9.8 7.6 6.7 <5 <5 <5 <5 <5

- 18 043669

20 10.0 7.7 6.4 <5 <5 <5 <5 <5 21 >10 7.8 6.4 <5 <5 <5 <5 <5 22 9.8 7.6 5.5 <5 <5 <5 <5 <5 23 9.4 7.2 5.9 <5 <5 <5 <5 <5 24 9.1 7.1 5.6 <5 <5 <5 <5 <5 25 9.7 7.4 6.2 <5 <5 <5 <5 <5 26 9.8 7.4 5.8 <5 <5 <5 <5 <5 27 >10 7.9 6.4 <5 <5 <5 <5 <5 28 >10 7.8 6.1 <5 <5 <5 5.3 <5 29 >10 7.6 6.4 <5 <5 <5 <5 <5 thirty 9.9 7.9 6.4 <5 <5 <5 <5 <5 31 9.9 7.6 5.9 <5 <5 <5 <5 <5 32 >10 7.7 5.8 <5 <5 <5 <5 <5 33 9.3 6.9 5.9 <5 <5 <5 <5 <5 34 >10 7.5 5.3 <5 <5 <5 <5 <5 35 >10 7.4 5.9 <5 <5 5.2 <5 <5 36 9.7 7.5 6.0 <5 <5.1 <5 <5 <5 37 9.2 6.5 5.7 <5 <5 <5 <5 <5 38 8.0 5.5 <5.1 <5 <5 <5 <5 <5 39 9.0 7.1 5.4 <5 <5 <5 <5 <5 40 9.5 6.8 5.8 <5 <5 <5.1 <5 <5 41 8.6 6.2 <5.1 <5 <5 <5 <5 <5 42 8.9 6.5 <5.1 <5 <5 <5 <5 <5 43 7.8 5.7 <5 <5 <5 <5 <5 <5 44 8.5 6.7 <5.1 <5 <5 <5 <5 <5 45 9.2 7.0 5.2 <5 <5 <5 <5 <5 46 8.1 6.2 5.4 <5 <5 <5 <5 <5 47 9.0 6.7 5.8 <5 <5 <5 <5 <5 48 9.0 6.5 5.9 <5 <5 <5 <5 <5 49 8.8 6.6 5.7 <5 <5 <5 <5 <5 50 8.4 6.4 5.2 <5 <5 <5 <5 <5 51 >10 7.9 6.4 <5 <5 <5 <5 <5 52 9.3 7.0 5.6 <5 <5 <5.1 <5 <5

Based on the measured data, it can be seen that the examples are DNA-PK inhibitors that are selective for these specific targets: TTK, JAK1, JAK2, JAK3, Aurora A, Aurora B. Comparison of pIC ₅₀ values for the enzymes indicates that the examples are characterized selectivity for DNA-PK that is >2.5 log units greater than that for the other targets shown. This equates to >300-fold selectivity between IC ₅₀ values.

Moreover, certain DNA-PK inhibitors of formula (I) exhibit a favorable volume of distribution and, as a consequence, may have a longer half-life in vivo, in particular when R ¹ represents a basic group, for example, where R ¹ represents pyrrolidinyl or piperidinyl. The longer half-life means that these compounds may be useful in certain combination therapy scenarios.

In table D shows pharmacokinetic data for Examples 44 and 52. Data were based on plasma collected within 24 hours of acute dosing of the compound.

- 19 043669 of formula (I) to either han/wistar rats or beagle dogs. Han/wistar rats were dosed

i) Example 44 as either a 1 mg/kg intravenous dose (100% deionized water adjusted to pH 3.96 with 1 M HCl) or a 5 mg/kg oral dose (5% DMSO/95% SBE-B -CD (30% w/v) in water adjusted to pH 3.87 with 1 M HCl); or ii) Example 52 as either an intravenous dose of 0.5 mg/kg (5% DMSO/95% SBE-B-CD (30% w/v) in water) or an oral dose of 1 mg/kg (100% (0.5% HPMC/0.1% TWEEN in water)).

Beagle dogs were given a dose

i) Example 44 (DMSO/95% SBE-B-CD (30% w/v) in water for both routes of administration) as either a 0.5 mg/kg intravenous dose or a 1 mg/kg oral dose ; or ii) Example 52 as either a 0.5 mg/kg intravenous dose (5% DMSO/95% SBE-B-CD (30% w/v) in water) or a 1 mg/kg oral dose ( 100% (0.5% HPMC/0.1% TWEEN in water)).

Table D

Example 44 Example 52 Selected PK indicators in rats: ground clearance (CL) 13 ml/min/kg 12.2 ml/min/kg volume of distribution (V) 3.4 l/kg 1.65 l/kg 1½ for oral dose 3.9 h 3.6 h 1½ for intravenous dose 4.3 h 4.6 h Individual RK- dog indicators: ground clearance (CL) 7.5 ml/min/kg 16 ml/min/kg volume of distribution (V) 7.8 l/kg 5.4 l/kg 1½ for oral dose undefined 6.3 h 1½ for intravenous dose 15 h 6.2 h

Compounds may further be selected based on additional biological or physical properties that can be measured using techniques known in the art and that can be used to evaluate or select compounds for therapeutic or prophylactic use.

Because of their DNA-PK inhibitory activity, the compounds of formula (I) and their pharmaceutically acceptable salts are expected to be useful in therapy.

Accordingly, the compounds of the present invention have value as antitumor agents. In particular, the compounds of the present invention are useful as antiproliferative, apoptotic and/or anti-invasive agents for containing and/or treating solid tumor and/or hematological malignancies. In particular, the compounds of the present invention are expected to be useful in the prevention or treatment of those tumors that are sensitive to DNA-PK inhibition. Additionally, the compounds of the present invention are expected to be useful in the prevention or treatment of those tumors that are fully or partially mediated by DNA-PK. Thus, the compounds can be used to provide an inhibitory effect on the DNA-PK enzyme in a warm-blooded animal in need of such treatment.

As stated herein, DNA-PK inhibitors should have therapeutic value for the treatment of proliferative diseases such as cancer and, in particular, solid tumors and their metastases, and various forms of hematological tumors.

Anticancer effects that are accordingly useful in treating cancer in a patient include, but are not limited to, antitumor effects, response rate, time to disease progression, and survival rate. The antitumor effects of the treatment method of the present invention include, but are not limited to, suppression of tumor growth, inhibition of tumor growth, tumor regression, reduction in tumor size, increased time to tumor regrowth upon cessation of treatment, and inhibition of disease progression. Anti-cancer effects include preventative treatment as well as treatment of existing disease.

When cancer is mentioned, it includes both non-metastatic cancer and metastatic cancer, whereby cancer treatment involves treating both primary tumors and metastatic tumors.

DNA-PK inhibitory activity refers to the reduction in DNA-PK activity as a direct or indirect response to the presence of a compound of formula (I) or a pharmaceutically acceptable salt thereof relative to DNA-PK kinase activity in the absence of a compound of formula (I) or a pharmaceutically acceptable salt thereof . This decrease in activity may be due to the direct interaction of a compound of formula (I) or a pharmaceutically acceptable salt thereof with DNA-PK or due to the interaction of a compound of formula (I) or a pharmaceutically acceptable salt thereof with one or more other factors, which in turn influence DNA-PK activity. For example, a compound of formula (I) or a pharmaceutically acceptable salt thereof may reduce the level of DNA-PK by directly binding to DNA-PK, by influencing (directly or indirectly) another factor to reduce DNA-PK activity, or by (directly or indirectly) reducing the amount of DNA-PK present in a cell or organism.

The term therapy is intended to have its ordinary meaning when referring to a disease, to denote the complete or partial relief of one, more or all of its symptoms or the elimination or relief of the pathology underlying it. The term therapy also includes prophylaxis unless specifically stated otherwise. The terms therapeutic and therapeutically should be interpreted accordingly.

The term prophylaxis is intended to have its ordinary meaning and includes primary prevention to prevent the development of a disease and secondary prevention in which the disease has already developed and the patient is temporarily or permanently protected from exacerbation or worsening of the disease or the development of new symptoms associated with the disease.

The term treatment is used as a synonym for the term therapy. Likewise, the term treat can be thought of as the application of a therapy, where the therapy is as defined herein.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in therapy.

In one embodiment, the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicinal product is provided.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of a DNA-PK mediated disease. In one embodiment, said DNA-PK mediated disease is cancer. In one embodiment, said cancer is a solid cancer or hematologic malignancy. In one embodiment, said cancer is selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, hematologic malignancy, non-small cell lung cancer, small cell lung cancer, gastric cancer, and squamous cell carcinoma of the head and neck.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer.

In one embodiment, the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for the manufacture of a medicament for the treatment of a disease mediated by DNA-PK. In one embodiment, said DNA-PK-mediated disease is cancer. In one embodiment, said cancer is a solid cancer or hematologic malignancy. In one embodiment, said cancer is selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, hematologic malignancy, non-small cell lung cancer, small cell lung cancer, gastric cancer, and squamous cell carcinoma of the head and neck.

In one embodiment, the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for the manufacture of a medicament for the treatment of cancer.

In one embodiment, a method of treating a disease benefiting from DNA-PK inhibition in a warm-blooded animal in need of such treatment is provided, which comprises administering to said warm-blooded animal a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, said disease is cancer. In one embodiment, said cancer is a solid cancer or hematologic malignancy. IN

- 21 043669 In one embodiment, said cancer is selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, hematological malignancy, non-small cell lung cancer, small cell lung cancer, gastric cancer and squamous cell carcinoma of the head and neck.

In one embodiment, a method of treating cancer in a warm-blooded animal in need of such treatment is provided, which comprises administering to said warm-blooded animal a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof.

The term therapeutically effective amount refers to the amount of a compound of formula (I) described in any of the embodiments herein that is effective to provide therapy in a subject or to treat a disease or disorder in a subject. In the case of cancer, a therapeutically effective amount may cause any of the changes observed or measured in the subject as described above in the definition of therapy, treatment and prevention. For example, an effective amount may reduce the number of cancer or tumor cells;

reduce overall tumor size;

suppress or stop the infiltration of tumor cells into peripheral organs, including, for example, soft tissue and bone;

suppress and stop tumor metastasis;

suppress and stop the growth of tumors;

relieve to some extent one or more symptoms associated with cancer;

reduce disease severity and mortality;

improve quality of life; or provide a combination of such effects.

An effective amount may be an amount sufficient to reduce the severity of symptoms of a disease sensitive to inhibition of DNA-PK activity. In the case of cancer therapy, in vivo effectiveness can be measured, for example, by assessing survival time, time to disease progression (TTP), response rates (RR), duration of response and/or quality of life. As recognized by those skilled in the art, effective amounts may vary depending on the route of administration, the use of the excipient, and the concomitant use of other agents. For example, if a combination therapy is used, the amount of a compound of formula (I) or a pharmaceutically acceptable salt described herein and the amount of other pharmaceutically active agent(s), if combined, are collectively effective for treating a target disorder in an animal patient. As used herein, combined amounts are a therapeutically effective amount, when combined, if they are sufficient to reduce the severity of symptoms of a disease sensitive to inhibition of DNA-PK activity as described above. Typically, such amounts can be determined by one skilled in the art, for example, based on the dosage range described herein for a compound of formula (I) or a pharmaceutically acceptable salt thereof, and the approved or otherwise published drug(s). dosage range(s) of the other pharmaceutically active compound(s).

Warm-blooded animals include, for example, humans.

In one embodiment, a method of treating cancer in a warm-blooded animal in need of such treatment is provided, which comprises administering to said warm-blooded animal a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof. In one embodiment, said cancer is a solid cancer or hematologic malignancy. In one embodiment, said cancer is selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, hematologic malignancy, non-small cell lung cancer, small cell lung cancer, gastric cancer, and squamous cell carcinoma of the head and neck.

In any embodiment, where cancer is referred to in a general sense, said cancer may be a solid cancer or a hematologic malignancy, such as breast cancer, ovarian cancer, pancreatic cancer, non-small cell lung cancer, small cell lung cancer, gastric cancer, and squamous cell carcinoma of the head and neck.

The anticancer treatment described herein may be useful as monotherapy or may include, in addition to administration of a compound of formula (I), conventional surgery, radiation therapy or chemotherapy, or a combination of such additional therapies. Such conventional surgery, radiation therapy or chemotherapy may be used simultaneously, sequentially or separately from treatment with a compound of formula (I).

When combination therapy is administered simultaneously, this involves treating the patient with a single dosage form (eg, tablet) containing both a compound of formula (I) or a pharmaceutically acceptable salt thereof and an additional anticancer agent; as well as the simultaneous administration of a dose of separate dosage forms, each of which separately contains one of the corresponding components of the combination.

If the combination therapy is administered sequentially or separately, it involves treating a patient with a first dosage form (eg, a tablet) containing a compound of formula (I) or a pharmaceutically acceptable salt thereof, and then treating the same patient with a second dosage form containing an additional anticancer agent. substance; or treating a patient with one dosage form (eg, a tablet) containing a particular anticancer agent and then treating the same patient with a second dosage form containing a compound of formula (I) or a pharmaceutically acceptable salt thereof.

The interval between successive or divided doses can be determined by the practitioner based on the information provided herein.

Radiation therapy may include one or more of the following treatment categories:

i) external radiation therapy, which uses electromagnetic radiation, and intraoperative radiation therapy, which uses electromagnetic radiation;

ii) internal radiotherapy or brachytherapy, including interstitial radiotherapy or intraluminal radiotherapy; or iii) systemic radiation therapy, including, but not limited to, iodine 131 and strontium 89 therapy.

Therefore, in one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof and radiation therapy are provided for use in the treatment of cancer. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with radiation therapy.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof and radiation therapy are provided for use in simultaneous, separate or sequential treatment of cancer. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered simultaneously, separately or sequentially with radiation therapy. In one embodiment, there is provided a method of treating cancer in a warm-blooded animal in need of such treatment, which comprises administering to said warm-blooded animal a compound of formula (I) or a pharmaceutically acceptable salt thereof and administering radiation therapy, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof and Radiation therapies are collectively effective in providing anticancer effects.

In one embodiment, there is provided a method of treating cancer in a warm-blooded animal in need of such treatment, which comprises administering to said warm-blooded animal a compound of formula (I) or a pharmaceutically acceptable salt thereof and simultaneous, separate or sequential application of radiation therapy, wherein the compound of formula (I) or its pharmaceutically acceptable salt and radiation therapy are collectively effective in providing an anticancer effect. In one embodiment, the cancer is a solid cancer.

In any embodiment, the radiation therapy is selected from the group consisting of one or more of the categories of radiotherapy listed under items (i)-(iii) above.

Chemotherapy may include one or more of the following categories of anticancer agents:

i) antiproliferative/antineoplastic drugs and their combinations used in the drug therapy of malignant tumors, such as antitumor antibiotics (for example, anthracyclines such as adriamycin, bleomycin, doxorubicin, liposomal doxorubicin, pyrarubicin, daunomycin, valrubicin, epirubicin, idarubicin, mitomycin- C, dactinomycin, amrubicin and mithramycin); and topoisomerase inhibitors (eg, epipodophyllotoxins such as etoposide and teniposide, amsacrine, irinotecan, topotecan and camptothecin);

ii) inhibitors of DNA repair mechanisms such as CHK kinase; ATM inhibitors (such as AZD0156 and AZD1390); poly(ADP-ribose) polymerase inhibitors (PARP inhibitors, including olaparib); ATR kinase inhibitors (such as cerelasertib/AZD6738) and WEE1 kinase inhibitors (such as adavosertib/AZD1775/MK-1775); and iii) immunotherapeutic approaches, including, for example, ex vivo and in vivo approaches to enhance the immunogenicity of a patient's tumor cells, such as PD-L1 blocking antibodies (eg, durvalumab/MEDI473 6).

Therefore, in one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one additional antitumor agent are provided for use in the treatment of cancer. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with an additional antitumor agent. In one embodiment, there is one additional antitumor agent. In one embodiment, there are two additional antitumor agents. In one embodiment, there are three or more additional antitumor agents.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one additional antitumor agent are provided for use in simultaneous, separate or sequential treatment of cancer. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered simultaneously, separately or sequentially with an additional antitumor agent.

In one embodiment, a method of treating cancer in a warm-blooded animal in need of such treatment is provided, which comprises administering to said warm-blooded animal a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one additional antitumor agent, wherein an amount of a compound of formula (I) or its pharmaceutically acceptable salt and the additional antitumor agent are collectively effective in providing an anticancer effect.

In one embodiment, there is provided a method of treating cancer in a warm-blooded animal in need of such treatment, which comprises administering to said warm-blooded animal a compound of formula (I) or a pharmaceutically acceptable salt thereof and simultaneously, separately or sequentially administering to said warm-blooded animal at least one additional antitumor agent wherein the amounts of a compound of formula (I) or a pharmaceutically acceptable salt thereof and the additional antitumor agent are collectively effective to produce an anticancer effect.

In any embodiment, the additional antitumor agent is selected from the group consisting of one or more antitumor agents listed in paragraphs (i)-(iii) above.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one antineoplastic agent are provided for use in the treatment of cancer. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with at least one antineoplastic agent. In one embodiment, the antineoplastic agent is selected from the list of antineoplastic agents in (i) above.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one antineoplastic agent are provided for use in simultaneous, separate or sequential treatment of cancer. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered simultaneously, separately or sequentially with at least one antineoplastic agent. In one embodiment, the antineoplastic agent is selected from the list of antineoplastic agents in (i) above.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one additional antitumor agent selected from the group consisting of doxorubicin or liposomal doxorubicin, olaparib, AZD6738 and AZD0156, for use in the treatment of cancer are provided. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with at least one additional antitumor agent selected from the group consisting of doxorubicin, liposomal doxorubicin, olaparib, AZD6738 and AZD0156.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one additional antitumor agent selected from the group consisting of doxorubicin or liposomal doxorubicin, olaparib, AZD6738 and AZD0156 are provided for use in simultaneous, separate or sequential treatment cancer. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered simultaneously, separately or sequentially with at least one additional antitumor agent selected from the group , consisting of doxorubicin, liposomal doxorubicin, olaparib, AZD6738 and AZD0156.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with olaparib. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered simultaneously, separately

- 24 043669 or sequentially with olaparib.

In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered in combination with doxorubicin or liposomal doxorubicin. In one embodiment, a compound of formula (I) or a pharmaceutically acceptable salt thereof is provided for use in the treatment of cancer, wherein the compound of formula (I) or a pharmaceutically acceptable salt thereof is administered simultaneously, separately or sequentially with doxorubicin or liposomal doxorubicin.

In one embodiment, a pharmaceutical composition is provided comprising a compound of formula (I) and at least one additional antitumor agent. In one embodiment, the pharmaceutical composition also contains at least one pharmaceutically acceptable excipient.

In one embodiment, a pharmaceutical composition is provided comprising a compound of formula (I) and at least one additional antitumor agent for use in the treatment of cancer. In one embodiment, the pharmaceutical composition also contains at least one pharmaceutically acceptable excipient.

According to a further embodiment, there is provided a kit containing

a) a compound of formula (I) or a pharmaceutically acceptable salt thereof in a first unit dosage form;

b) another additional antitumor agent in an additional unit dosage form;

c) container means for containing the said first and additional standard dosage forms; and optional

d) instructions for use.

In one embodiment, the antineoplastic agent comprises an antineoplastic agent.

In any embodiment where an antineoplastic agent is mentioned, the antineoplastic agent is one or more of the agents listed in (i) above.

The compounds of formula (I) and their pharmaceutically acceptable salts can be administered in the form of pharmaceutical compositions containing one or more pharmaceutically acceptable excipients.

Therefore, in one embodiment, a pharmaceutical composition is provided comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient.

The compositions may be in a form suitable for oral administration (for example, tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or tinctures), for topical application (for example, creams , ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example, as a fine powder or liquid aerosol), for administration by insufflation (for example, as a fine powder), or for parenteral administration (for example, as sterile aqueous or oily solution for intravenous, subcutaneous or intramuscular administration of the dose) or in the form of a suppository for rectal administration of the dose. The compositions can be prepared by conventional procedures using conventional pharmaceutical excipients well known in the art. Thus, compositions intended for oral administration may contain, for example, one or more coloring agents, sweeteners, flavoring agents and/or preservatives.

In one embodiment, a pharmaceutical composition is provided comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient for use in therapy.

In one embodiment, a pharmaceutical composition is provided comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient for use in the treatment of cancer. In one embodiment, said cancer is a solid cancer or hematologic malignancy. In one embodiment, said cancer is selected from the group consisting of breast cancer, ovarian cancer, pancreatic cancer, hematologic malignancy, non-small cell lung cancer, small cell lung cancer, gastric cancer, and squamous cell carcinoma of the head and neck.

Normally, a compound of formula (I) will be administered to a warm-blooded animal at a standard dose within the range of 2.5-5000 mg/m ² body surface area of the animal, i.e. approximately 0.05-100 mg/kg, and this amount normally provides a therapeutically effective dose. A unit dosage form such as a tablet or capsule will typically contain, for example, 0.1-250 mg of active ingredient. The daily dose will necessarily vary depending on the subject receiving treatment,

- 25 043669 the specific route of administration, any co-administered therapeutic agents and the severity of the disease being treated.

Examples

Various embodiments are illustrated through the following examples. The embodiment should not be interpreted as limited to the examples.

Unless otherwise stated, starting materials were commercially available. All solvents and commercial reagents were of laboratory grade and were used as received.

General information about experiments.

In the process of obtaining examples, the following was generally done.

(i) The operations were carried out at room temperature (RT), i.e. in the range from 17 to 25°C, and under an atmosphere of an inert gas such as N ₂ or Ar, unless otherwise indicated.

(ii) Typically, the progress of the reactions was monitored using thin layer chromatography (TLC) and/or analytical high performance liquid chromatography (HPLC or UPLC), which is usually coupled to a mass spectrometer (LCMS); The reaction times given are not necessarily the minimum achievable.

(iii) If necessary, organic solutions were dried over anhydrous MgSO ₄ or Na ₂ SO ₄ , work-up procedures were carried out using conventional phase separation techniques or by using SCX as described in (xiii), evaporation operations were carried out either by rotary evaporation in vacuo, or in Genevac HT-4/EZ-2 or Biotage V10.

(iv) Yields, when given, are not necessarily the maximum achievable and reactions were repeated as necessary if more product was required to be obtained.

(v) Typically, the structures of the final products of formula (I) were confirmed by nuclear magnetic resonance (NMR) and/or mass spectroscopy techniques; Electrospray mass spectroscopy data were typically acquired using a Waters Acquity UPLC coupled to a Waters single quadrupole detector mass spectrometer acquiring data for both positive and negative ions, and typically only ions relevant to the parent structure were reported; chemical shift values for proton NMR were measured on the delta scale using either a Bruker AV500 spectrometer operating at a field strength of 500 MHz, a Bruker AV400 operating at 400 MHz, or a Bruker AV300 operating at 300 MHz. Unless otherwise stated, NMR spectra were obtained at 500 MHz in d6-dimethyl sulfoxide. The following abbreviations were used: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad; qn, quintet.

(vi) Unless otherwise stated, compounds containing asymmetric carbon and/or sulfur atoms were not separated.

(vii) The intermediates were not necessarily fully purified, but their structures and purity were assessed by TLC, analytical HPLC/UPLC, and/or NMR analysis, and/or mass spectrometry.

(viii) Unless otherwise stated, flash column chromatography (fcc) was performed on Merck Kieselgel silica (No. 9385), or reverse phase chromatography silica (Fluka 90 C18 silica gel), or Silicycle cartridges (40 silica -63 µm, weight 4-330 g), or on Grace Resolv cartridges (4-120 g), or on RediSep Rf 1.5 Flash columns, or on high-efficiency RediSep Rf Gold Flash columns (weight 150-415 g), or on RediSep Rf Gold C18 reverse phase chromatography columns (silica 20-40 µm) either manually or automatically using an Isco CombiFlash Companion or similar system.

(ix) Preparative reverse phase HPLC (RP HPLC) was performed on C18 silica for reverse phase chromatography, typically using a Waters XSelect CSH C18 column (5 µm silica, 30 mm diameter, 100 mm length) using mixtures with decreasing polarity as eluents, for example [containing 0.1% formic acid or 0.3-5% aqueous ammonium hydroxide (d=0.91)] as solvent A and acetonitrile as solvent B; A typical procedure would be: solvent gradient over 10-20 min at a flow rate of 40-50 ml/min from a 95:5 mixture of solvents A and B, respectively, to a 5:95 mixture of solvents A and B (or an alternative ratio if necessary).

(x) The following analytical UPLC methods were used; Typically, C18 silica was used for reverse phase chromatography at a flow rate of 1 ml/min and detection was carried out using electrospray mass spectrometry and UV absorbance detection at wavelengths in the range 220-320 nm. Analytical UPLC was performed on CSH C18 silica reverse phase chromatography using a Waters XSelect CSH C18 2.1 x 50 mm 1.7 micron particle size column. Gradient analysis was used using mixtures of decreasing polarity as eluent, for example mixtures of decreasing polarity water (containing 0.1% formic acid or 0.1% ammonia) as solvent A and acetonite.

- 26 043669 rila as solvent B. A typical 2 min analytical UPLC method would use a solvent gradient of 1.3 min, at a flow rate of approximately 1 ml/min, from a 97:3 mixture of solvents A and B, respectively, to mixture 3: 97 solvents A and B.

(xi) When certain compounds were prepared as an acid addition salt, such as a monohydrochloride salt or a dihydrochloride salt, the stoichiometric composition of the salt was based on the number and nature of the main groups in the compound, and the exact stoichiometric composition of the salt was not usually determined, for example, by elemental data analysis.

(xii) When reactions refer to the use of microwave processing, one of the following microwave reactors was used: Biotage Initiator, Personal Chemistry Emrys Optimizer, Personal Chemistry Smithcreator, or CEM Explorer.

(xiii) Compounds were purified by strong cation exchange chromatography (SCX) using Isolute SPE Flash SCX-2 or SCX-3 columns (International Sorbent Technology Limited, MidGlamorgan, UK).

(xiv) The following preparative chiral HPLC methods were performed using Gilson GX-281 HPLC and DAICEL CHIRALPAK IC (2x25 cm, 5 µm), DAICEL CHIRALPAK IF (2x25 cm, 5 µm) or XBridge Prep OBD C18 column (3x15 cm, 5 µm ); Typically, the flow rate was 10-350 ml/min, and detection was carried out using UV absorbance at a typical wavelength of 254 nm. A sample concentration of approximately 1-100 mg/mL in a suitable solvent mixture was used, with an injection volume of 0.5-10 mL, a run time of 10-150 min, and a typical oven temperature of 25-35°C.

(xv) The following analytical chiral HPLC methods were performed using Shimadzu UFLC and Daicel CHIRALPAK IC-3 (50x4.6 mm, 3 µm) or Daicel CHIRALPAK IF-3 (50x4.6 mm, 3 µm); Typically the flow rate was 1 ml/min and detection was performed using UV absorbance at a typical wavelength of 254 nm. A sample concentration of approximately 1 mg/mL in a suitable solvent such as EtOH was used, with an injection volume of approximately 10 μL, a run time of 10-60 min, and a typical oven temperature of 25-35°C.

(xvi) The following preparative chiral supercritical fluid chromatography (SFC) methods were used; Typically, the flow rate was approximately 70 ml/min and detection was carried out using UV absorbance at a typical wavelength of 254 nm. A sample concentration of approximately 100 mg/mL in a suitable solvent such as MeOH was used, with an injection volume of approximately 0.5 mL, a run time of 10-150 min, and a typical oven temperature of 25-35°C.

(xvii) Typically, examples and intermediates were named using ACD Name, the Structure to Name portion of ChemDraw Ultra (CambridgeSoft) or Biovia Draw 2016.

(xviii) In addition to those above, the following abbreviations have been used:

ASGO acetic anhydride HPLC high performance liquid chromatography BINAP 2,2'-bis(diphenylphosphino)1,G-binaphthalene iPrOH iso-propanol CDCL deuterated chloroform MeCN acetonitrile END concentrated Mel iodomethane DBU 1.8- diazabicyclo[5,4,0]undec- 7-en MeOD O4-methanol DCM dichloromethane MeOH methanol

- 27 043669

DIPEA AA-diisopropylethylamine MTBE methyl threot butyl ether DMA Α,Α-dimethylacetamide mass/charge mass spectrometric peak(s) DMAP 4-dimethylaminopyridine NaH sodium hydride DME 1,2-dimethoxyethane N.B.S. A-bromosuccinimide DMF Α,Α-dimethylformamide NH4CI ammonium chloride DMF- DMA dimethylformamide- dimethyl acetal NMP 1-methylpyrrolidin-2-one DMSO dimethyl sulf oxide k.t. room temperature DSC differential scanning calorimetry sat. saturated Et _3N triethylamine SCX strong cation exchange EtOAc ethyl acetate SFC supercritical fluid chromatography Et ₂ O diethyl ether TV AB tetra-n-butylammonium bromide EtOH ethanol TBAF tetra-n-butylammonium fluoride F.A. formic acid THF tetrahydrofuran fee flash column chromatography XRPD X-ray powder diffraction 4 watch) RuPhos 2-d icyclohexylphosphino-26'-diisopropoxybiphenyl RuPhos Pd G3 (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,G-biphenyl)[2-(2'amino-1,1'-biphenyl)]palladium(P) methanesulfonate Brettphos Pd G3 [(2-dicyclohexylphosphino-3,6-dimethoxy-2',4',6'-triisopropyl-1, Gbiphenyl)-2-(2'-amino-1,1'-biphenyl)] palladium(II) methanesulfonate methanesulfonate

(xix) For XRPD analysis, a Bruker D4 instrument was used. The X-ray powder diffraction pattern was determined by mounting a sample of crystalline material on a Bruker single crystal silicon (SSC) plate holder and spreading the sample as a thin layer using a glass slide. The sample was spun at 30 rpm (to improve counting statistics) and irradiated with X-rays generated by a long fine-focus copper tube operating at 40 kV and 40 mA, at a wavelength of 1.5418 angstroms. The collimated X-ray source was passed through an auto-divergence slit mounted on V20, and the reflected radiation was directed through a 5.89 mm anti-scatter slit and a 9.55 mm detector slit. Samples were measured for reflection geometry in a θ - 2θ configuration over a scan range of 2 to 40° 2Θ at a nominal exposure of 0.12 s in 0.02° increments. The device was equipped with a coordinate-sensitive detector (Lynxeye). Those skilled in the art of powder X-ray diffraction will recognize that the relative intensity of the peaks can be influenced, for example, by grains larger than 30 microns and aspect ratios far from unity, which can affect the analysis of the samples. One skilled in the art will also appreciate that the position of the reflections can be influenced by the precise height at which the sample is placed in the diffractometer and the calibration of the diffractometer zero. The flatness of the sample surface may also have a minor effect. Therefore, the XRD data presented should not be taken as absolute values.

(xx) In the case of differential scanning calorimetry, a TA Instruments Q2000 DSC device was used. Typically less than 3 mg of material contained in a standard aluminum cookware

- 28 043669 lid, heated in the temperature range from 25 to 300°C at a constant heating rate of 10°C/min. A nitrogen purge gas was used at a flow rate of 50 ml/min.

Temperature data was analyzed using standard software, such as Universal V.4.5A from TA INSTRUMENTS®.

Intermediate 1: 5-amino-2-bromo-4-methylbenzoic acid.

NBS (11.87 g, 66.15 mmol) was added portionwise to 3-amino-4-methylbenzoic acid (10.00 g, 66.15 mmol) in DMF (50 ml) at 5° C. over a period of 5 min so that the temperature does not rise above 15°C. The resulting solution was stirred at 5°C for 1 hour. The reaction mixture was poured onto ice water (250 ml) with stirring. The resulting solid was filtered, washed with ice water and dried to give the title compound (15.21 g, 100%) as a pale pink solid.

^1H NMR (400 MHz, DMSO) 2.04-2.09 (3H, m), 5.21 (2H, s), 7.06 (1H, s), 7.21 (1H, d), 12 .84 (1H, s).

Intermediate 2: Methyl 5-amino-2-bromo-4-methylbenzoate.

Thionyl chloride (4.80 ml, 66.11 mmol) was added dropwise to 5-amino-2-bromo-4-methylbenzoic acid (15.21 g, 66.11 mmol) in MeOH (200 ml) at 20°C in over a period of 5 minutes. The resulting solution was stirred at reflux for 2 hours. The reaction mixture was allowed to cool, quenched with a small amount of water, and the solvent was removed in vacuo. The reaction mixture was diluted with EtOAc (250 ml) and washed successively with saturated NaHCO ₃ (100 ml), water (100 ml) and brine (100 ml). The organic layer was passed through a phase separation filter paper and evaporated to give the title compound (14.85 g, 92%) as a red oil.

1H NMR (400 MHz, DMSO) 2.06-2.11 (3H, m), 3.80 (3H, s), 5.28 (2H, s), 7.06 (1H, s), 7. 22-7.28 (1H, m); mass/charge MN ⁺ 244.

Intermediate 3: Methyl 2-bromo-5-((tert-butoxycarbonylamino))-4-methylbenzoate.

Di-tert-butyl dicarbonate (20.40 g, 93.47 mmol) was added to methyl 5-amino-2-bromo-4methyl benzoate (15.21 g, 62.31 mmol) in ethanol (125 ml) at room temperature The solution was stirred at room temperature for 18 hours.An additional 0.5 eq. di-tert-butyl dicarbonate (6.80 g, 31.15 mmol) and the reaction mixture was stirred at room temperature for an additional 18 hours. EtOH was removed in vacuo and the resulting suspension was diluted with n-heptane (250 ml). The solid was filtered, washed with n-heptane and dried to give the title compound (14.82 g, 69%) as a gray solid.

^1H NMR (400 MHz, DMSO) 1.48 (9H, s), 2.25 (3H, s), 3.84 (3H, s), 7.56-7.6 (1H, m), 7 .89 (1H, s), 8.73 (1H, s);

mass/charge MN ⁺ 344.

Intermediate 4: tert-butyl (4-bromo-5-formyl-2-methylphenyl)carbamate.

Di-iso-butylaluminum hydride (1 M, 100 mL, 100.0 mmol) was added to methyl 2-bromo-5-((tert-butoxycarbonyl)amino)-4-methylbenzoate (11.47 g, 33.33 mmol) in DCM (200 ml) at -78°C. Solution ne

- 29 043669 stirred at -78°C for 1 hour. Methanol (20 ml) was added slowly to stop the reaction and the solution was heated to room temperature. The reaction mixture was diluted with 0.5 M aq. HCl (250 ml) and diethyl ether (250 ml). The organic phase was isolated, washed with brine (200 ml), passed through filter paper to separate the phases and the solvent was removed in vacuo to obtain a mixture of the desired aldehyde and alcohol. Manganese(IV) oxide (10.78 g, 124.1 mmol) was added in one portion to the mixture in DCM (260 ml) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. An additional 5 equivalents of oxide were added manganese(IV) (14.48 g, 166.7 mmol) and the reaction mixture was stirred at RT for 1 hour. The reaction mixture was filtered through Celite®, washed with DCM (500 ml) and the solvent was removed in vacuo with obtaining the title compound (9.97 g, 96%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.48 (9H, s), 2.29 (3H, s), 7.63-7.66 (1H, m), 7.95 (1H, s), 8 .78 (1H, s), 10.14 (1H, s);

mass/charge MN ⁺ 314.

Intermediate 5: tert-butyl-N-[4-bromo-5-[(Z)-2-bromovinyl]-2-methyl-phenyl]carbamate.

HN^O./

O I

Potassium 2-methylpropan-2-olate (4.18 g, 37.24 mmol) was added portionwise to (bromomethyl)triphenylphosphonium bromide (16.24 g, 37.24 mmol) in THF (500 ml) at -78°C. To the resulting suspension was added dropwise tert-butyl-(4-bromo-5-formyl-2-methylphenyl)carbamate (9.75 g, 31.03 mmol) in THF (100 ml), the mixture was stirred for 16 hours and given let it slowly warm up to room temperature. N-heptane (500 ml) was added and the precipitate was filtered through celite. The solvent was removed in vacuo to obtain a light brown solid. The product was purified by fcc, eluting with 0-10% ethyl acetate in n-heptane to give the title compound (9.12 g, 75%) as a white solid.

1H NMR (400 MHz, DMSO) 1.47 (9H, s), 2.21 (3H, s), 6.87 (1H, d), 7.20 (1H, d), 7.51 (1H, s), 7.74 (1H, s), 8.62 (1H, s);

An ion with the required mass was not found for this intermediate.

Intermediate 6: Diethyl 6-((tert-butoxycarbonyl)amino)-7-methylcinnoline-1,2dicarboxylate.

Tert-butyl-N-[4-bromo-5-[(Z)-2-bromovinyl]-2-methyl-phenyl]carbamate (9.12 g, 23.32 mmol) was dissolved in 1,4-dioxane (150 ml) at room temperature in a nitrogen atmosphere. The reaction mixture was degassed by bubbling nitrogen through the mixture for 5 minutes. Add potassium carbonate (8.06 g, 58.30 mmol), diethylhydrazine-1,2-dicarboxylate (6.16 g, 34.98 mmol), N1,N2-dimethylethane-1,2-diamine (1.255 ml, 11 .66 mmol) and copper(I) iodide (1.110 g, 5.83 mmol) and the resulting dark green suspension was stirred at 100°C for 18 hours. The reaction mixture was allowed to cool to room temperature, filtered, washed with DCM (400 ml) and the filtrate was concentrated in vacuo. The product was purified by fcc eluting with 0-25% ethyl acetate in n-heptane to give the title compound (6.63 g, 70%) as a white foam.

¹ NMR (400 MHz, DMSO) 1.21 (6H, dt), 1.47 (9H, s), 2.22 (3H, s), 4.18 (4H, dq), 6.30 (1H, d), 7.11 (1H, s), 7.19 (1H, s), 7.25 (1H, s), 8.57 (1H, s);

mass/charge MN ⁺ 406.

- 30 043669

Intermediate 7: tert-butyl (7-methylcinnolin-6-yl)carbamate.

M aq. NaOH (7.95 mL, 15.91 mmol) was added to diethyl 6-((tert-butoxycarbonyl)amino)-7methylcinnoline-1,2-dicarboxylate (2.15 g, 3.18 mmol) in EtOH (25 mL ) at room temperature. The reaction mixture was stirred at room temperature for 18 hours, then concentrated in vacuo. The reaction mixture was diluted with EtOAc (100 ml) and the organic layer was isolated and washed successively with water (50 ml) and sat. saline solution (50 ml). The organic layer was passed through a phase separation filter paper, concentrated in vacuo, then purified by fcc, eluting with 0-100% EtOAc in n-heptane to give the title compound (0.370 g, 45%) as an orange foam.

1H NMR (400 MHz, DMSO) 1.53 (9H, s), 2.54 (3H, d), 8.09 (1H, dd), 8.24 (2H, d), 8.89 (1H, s), 9.19 (1H, d);

mass/charge MH+ 260.

Intermediate 8: 7-methylcinnolin-6-amine.

M HCl in 1,4-dioxane (1.54 ml, 6.17 mmol) was added to tert-butyl-(7-methylcinnolin-6yl)carbamate (320 mg, 1.23 mmol) in MeOH (5 ml) at and the reaction mixture was stirred at room temperature for 18 hours, then concentrated in vacuo to give the title compound (245 mg, 100%) as a bright orange solid.

1H NMR (400 MHz, DMSO) 2.42 (3H, d), 6.96 (1H, s), 7.91 (2H, s), 8.10 (1H, s), 8.22 (1H, d), 8.86 (1H, d);

mass/charge MH+ 160.

Intermediate 9: N-(7-methyl-6-quinolyl)-1,1-diphenylmethanimine.

ΪΎ J

Ph^Ph

Pd ₂ (dba) ₃ (0.918 g, 1.13 mmol) and sodium tert-butoxide (6.49 g, 67.54 mmol) were added to the degassed suspension of 6-bromo-7-methylquinoline (10 g, 45.03 mmol ), diphenylmethanimine (8.31 ml, 49.53 mmol) and rac-BINAP (1.40 g, 2.25 mmol) in toluene (172 ml). The reaction mixture was heated at 90°C for 1 hour. The reaction mixture was allowed to cool to room temperature, diluted with EtOAc (200 ml) and washed with water (100 ml). The aqueous layer was extracted with EtOAc (2x100 ml), then the combined organic layers were passed through filter paper to separate the phases and the solvent was removed in vacuo. The crude product was purified by fcc eluting with 0-100% EtOAc in n-heptane to give the title compound (14.30 g, 99%) as an orange solid.

1H NMR (400 MHz, DMSO) 2.35-2.4 (3H, m), 6.92 (1H, s), 7.20 (2H, dd), 7.25-7.34 (4H, m ), 7.49-7.55 (2H, m), 7.58 (1H, ddd), 7.72-7.79 (3H, m), 7.94 - 8.02 (1H, m), 8.66 (1H, dd);

mass/charge MH+ 323.

Intermediate 10: 7-methylquinoline-6-amine.

M aq. a solution of HCl (93.0 mL, 186.1 mmol) was added to N-(7-methyl-6-quinolyl)-1,1-diphenylmethanimine (15.00 g, 46.52 mmol) in THF (35 mL) and the reaction mixture was stirred for 1 hour. The reaction mixture was diluted with EtOAc (100 ml) and the layers were separated. The aqueous layer was further extracted with EtOAc (50 ml). The aqueous phase was neutralized with 2 M aq. The NaOH and the resulting solid were collected by filtration, washed with a small volume of water and dried to give the title compound (6.70 g, 91%) as a cream solid.

- 31 043669

1H NMR (400 MHz, DMSO) 2.25-2.32 (3H, m), 5.35 (2H, s), 6.87 (1H, s), 7.22 (1H, dd), 7. 61 (1H, s),

7.87-7.98 (1H, m), 8.46 (1H, dd);

mass/charge MN ⁺ 159.

Intermediate 11: 6-bromo-4-methoxy-7-methylquinoline.

Sodium methanolate (221 mg, 4.09 mmol) was added in one portion to 6-bromo-4-chloro-7-methylquinoline (350 mg, 1.36 mmol) in methanol (8 ml) at room temperature. The reaction mixture was stirred at 65°C for 1 day, then allowed to cool to room temperature. The reaction mixture was diluted with EtOAc (75 ml) and washed with water (20 ml) and sat. saline solution (20 ml). The organic phase was dried over MgSO ₄ , filtered and concentrated in vacuo to give the title compound (275 mg, 80%) as a cream solid.

1H NMR (400 MHz, DMSO) 2.54 (3H, s), 4.05 (3H, s), 7.03 (1H, d), 7.94 (1H, s), 8.30 (1H, s), 8.74 (1H, d);

mass/charge MN ⁺ 252.

Intermediate 12: 4-methoxy-7-methylquinoline-6-amine.

Brettphos G3 precatalyst (126 mg, 0.14 mmol) was added in one portion to 6-bromo-4-methoxy-7methylquinoline (350 mg, 1.39 mmol), cesium carbonate (905 mg, 2.78 mmol) and 0.5 M ammonia in 1,4-dioxane (5.55 ml, 2.78 mmol) in dioxane (9 ml) at room temperature. The reaction mixture was heated in a microwave reactor at 100°C for 3 days. The reaction mixture was filtered and the solid was washed with DCM (50 ml). The organic layers were combined and concentrated in vacuo, then purified by fcc, eluting with 0-5% MeOH in DCM to give the title compound (200 mg, 77%) as a white solid.

1H NMR (400 MHz, DMSO) 2.27 (3H, s), 3.96 (3H, s), 5.30 (2H, s), 6.75 (1H, d), 7.15 (1H, s), 7.55 (1H, s), 8.33 (1H, d);

mass/charge MN ⁺ 189.

Intermediate 13: N-(4-chloro-7-methyl-6-quinolyl)-1,1-diphenylmethanimine.

Pd2(dba) ₃ (23.8 mg, 0.03 mmol) and sodium tert-butoxide (169 mg, 1.75 mmol) were added to a degassed suspension of 6-bromo-4-chloro-7-methylquinoline (300 mg, 1 .17 mmol), diphenylmethanimine (216 μl, 1.29 mmol) and rac-BINAP (36.4 mg, 0.06 mmol) in toluene (4.46 ml). The reaction mixture was heated at 90°C for 1 hour, then allowed to cool to room temperature, diluted with EtOAc and washed with water. The aqueous layer was extracted with EtOAc, then the combined organic layers were dried over MgSO ₄ , concentrated in vacuo and purified by fcc eluting with 0-25% EtOAc in n-heptane to give the title compound (408 mg, 98%) in the form of a yellow resin.

^1H NMR (400 MHz, DMSO) 2.43 (3H, s), 7.14 (3H, s), 7.23 (3H, s), 7.28 (1H, d), 7.38-7 .56 (3H, m), 7.84 (3H, d), 8.55 (1H, d);

mass/charge MN ⁺ 357.

Intermediate 14: N-(4,7-dimethyl-6-quinolyl)-1,1-diphenylmethanimine.

The PdCl ₂ (dppf)-CH ₂ Cl ₂ adduct (37.3 mg, 0.05 mmol) was added in one portion to the degassed mixture of 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane ( 0.192 ml, 1.37 mmol), N-(4-chloro-7-methyl-6-quinolyl)-1,1diphenylmethanimine (326 mg, 0.91 mmol) and cesium carbonate (595 mg, 1.83 mmol) in 1,4-dioxane (12 ml) at room temperature. The resulting suspension was stirred in a capped flask at 120°C for 15 hours. The reaction mixture was allowed to cool to room temperature and filtered, then evaporated to dryness and purified by fcc, eluting with 0-40% EtOAc in n-heptane to give the title compound (254 mg, 83%) as a yellow gum.

1H NMR (400 MHz, DMSO) 2.38 (3H, d), 2.43 (3H, d), 6.86 (1H, s), 6.94-7.07 (1H, m), 7. 13 (2H, dd), 7.18-7.24 (3H, m), 7.38-7.59 (3H, m), 7.74-7.94 (3H, m), 8.55 ( 1H, d);

- 32 043669 mass/charge MH+ 337.

Intermediate 15: 4,7-dimethylquinoline-6-amine.

M aq. a solution of HCl (0.092 ml, 3.02 mmol) was added to N-(4,7-dimethyl-6-quinolyl)-1,1diphenylmethanimine (254 mg, 0.75 mmol) in THF (2 ml) and the reaction mixture was stirred in for 1 hour. Water was added to the mixture (to dissolve the solid) and the solution was loaded onto a 10 g SCX-2 column. Benzophenone was washed with MeOH (2x column volume), then the column contents were eluted with 1 M NH3 in MeOH to give the title compound (129 mg, 99%) as a cream solid.

1H NMR (400 MHz, DMSO) 2.29 (3H, d), 2.52 (3H, s), 5.37 (2H, s), 7.00 (1H, s), 7.04-7, 13 (1H, m), 7.51-7.66 (1H, m), 8.33 (1H, d);

mass/charge MH+ 173.

Intermediate 16: 6-chloro-7-nitroquinoxaline.

Oxalaldehyde (40% in water, 4.26 ml, 37.32 mmol) was added to 4-chloro-5-nitrobenzene-1,2-diamine (5.00 g, 26.65 mmol) in ethanol (100 ml) at The resulting solution was refluxed for 1 hour. The reaction mixture was cooled to rt and the resulting precipitate was filtered and dried to give the title compound (4.50 g, 81%) as a brown solid. substances.

1H NMR (400 MHz, DMSO) 8.57 (1H, s), 8.91 (1H, s), 9.15 (1H, d), 9.17 (1H, d);

mass/charge MH+ 210.

Intermediate 17: 6-methyl-7-nitroquinoxaline.

2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (2.97 ml, 21.27 mmol) was added to 6-chloro-7nitroquinoxaline (3.43 g, 16.37 mmol) in 1,4-dioxane (50 ml) and water (5 ml) at room temperature. Add K ₂ CO ₃ (6.79 g, 49.10 mmol) and dichloro-1,1'-bis(diphenylphosphino)ferrocene palladium(II) (1.197 g, 1.64 mmol) and the reaction mixture was refluxed for 2 hours. The reaction mixture was allowed to cool to room temperature, diluted with EtOAc (100 ml), washed with water (50 ml) and sat. brine (50 ml) and the organic layer was filtered through phase separation filter paper and concentrated in vacuo to give the crude product (2.50 g, 85% if pure), which was directly used in the next step.

^1H NMR (400 MHz, DMSO) 2.65 (3H, s), 8.19 (1H, s), 8.66 (1H, s), 9.04 (2H, dd);

mass/charge MH+ 190.

Intermediate 18: 7-methylquinoxaline-6-amine.

Iron (4.43 g, 79.29 mmol) and ammonium hydrochloride (0.707 g, 13.22 mmol) were added to 6-methyl-7nitroquinoxaline (2.50 g, 13.22 mmol) in EtOH (85 ml) and water (15 ml) at room temperature. The resulting mixture was heated at 100°C for 1 hour. The reaction mixture was allowed to cool to room temperature, filtered and washed with EtOH. The filtrate was concentrated in vacuo and the crude product was purified by fcc, eluting with 0-5% MeOH in DCM to give the title compound (1.80 g, 86%) as a yellow solid.

1H NMR (400 MHz, DMSO) 2.24-2.38 (3H, m), 5.83 (2H, s), 7.02 (1H, s), 7.61-7.68 (1H, m ), 8.44 (1H, d), 8.57 (1H, d);

mass/charge MH+ 160.

Intermediates 19 and 20: 7-chloro-2-methyl-6-nitroquinoxaline (major) and 6-chloro-2methyl-7-nitroquinoxaline (minor).

2-Oxopropanal (4.80 g, 26.65 mmol) was added to 4-chloro-5-nitrobenzene-1,2-diamine (5.00 g, 26.65 mmol) in EtOH (100 ml) at RT The resulting solution was heated in a reflux flask for 1 hour. The reaction mixture was allowed to cool to room temperature and the resulting precipitate was isolated by filtration and dried in vacuo to give a 6:1 mixture of 7-chloro-2-methyl -6-nitroquinoxaline (basic) and 6-chloro-2-methyl-7-nitroquinoxaline (4.67 g, 78%, common to both isomers).

^1H NMR (400 MHz, DMSO, main isomer) 2.78 (3H, s), 8.43 (1H, s), 8.84 (1H, s), 9.05 (1H, s);

mass/charge [M-N] ^- 222.

Intermediates 21 and 22: 2,7-dimethyl-6-nitroquinoxaline (major) and 2,6-dimethyl-7nitroquinoxaline (minor).

2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (1.60 ml, 11.40 mmol) was added to a 6:1 mixture of 7-chloro2-methyl-6-nitroquinoxaline and 6- chloro-2-methyl-7-nitroquinoxaline (2.04 g, 9.12 mmol) in 1,4-dioxane (50 ml) and water (5 ml) at room temperature. Potassium carbonate (3.15 g, 22.81 mmol) and dichloro-1,1'bis(diphenylphosphino)ferrocene palladium (II) (Pd106) (0.57 g, 0.76 mmol) and the reaction mixture was refluxed for 2 hours. the mixture was cooled, diluted with EtOAc (100 ml) and filtered. The filtrate was washed with water (50 ml) and sat. saline solution (50 ml). The organic layer was isolated, filtered through phase separation filter paper, concentrated in vacuo and purified by fcc eluting with 0-50% EtOAc in n-heptane to give the title compounds as a 6:1 mixture of isomers (1.01 g, 65%, total) as a tan-colored solid.

^1H NMR (400 MHz, DMSO, main isomer) 2.69 (3H, s), 2.76 (3H, s), 8.10 (1H, s), 8.66 (1H, s), 8. 98(1H, s);

mass/charge MN ⁺ 204.

Intermediates 23 and 24: 2,7-dimethylquinoxaline-6-amine (major) and 3,7-dimethylquinoxaline-6-amine (minor).

Iron (1.43 g, 25.69 mmol) and ammonium hydrochloride (229 mg, 4.28 mmol) were added to a 6:1 mixture of 2,7-dimethyl-6-nitroquinoxaline and 3,7-dimethylquinoxaline-6-amine ( 1.01 g, 4.28 mmol) in EtOH (30 ml) and water (5 ml) at room temperature under nitrogen. The resulting mixture was stirred at 100°C for 1 hour. The reaction mixture was cooled, filtered and washed with EtOH. The solvent was removed in vacuo and the crude product was purified by fcc, eluting with a gradient of 0 to 5% MeOH in DCM to give the title compounds (600 mg, 81%) as a brown solid (~5:1 mixture).

^1H NMR for main isomer (400 MHz, DMSO) 2.30 (3H, s), 2.56 (3H, s), 5.63 (2H, s), 7.01 (1H, s), 7. 56 (1H, s), 8.50 (1H, s);

mass/charge MH+ 174.

Intermediate 25: 5-chloroquinazoline-6-amine.

N-chlorosuccinimide (4.47 g, 33.48 mmol) was added in one portion to quinazoline-6-amine (4.86 g, 33.48 mmol) in DCM (150 ml) at 25°C. The resulting solution was stirred at 25°C for 16 hours. The solvent was removed under reduced pressure and the residue was purified by fcc, eluting with a gradient of 0 to 30% EtOAc in petroleum ether. The pure fractions were evaporated to dryness to give the title compound (6.30 g, 105%) as a pale yellow solid.

^1H NMR (400 MHz, DMSO) 6.31 (2H, s), 7.60 (1H, d), 7.76 (1H, d), 9.04 (1H, s), 9.41 (1H , s); mass/charge MN ⁺ 180.

Intermediate 26: 7-bromo-5-chloroquinazoline-6-amine.

NBS (5.00 g, 28.09 mmol) was added in one portion to 5-chloroquinazolin-6-amine (6.10 g, 33.96 mmol) in DCM (150 mL) at room temperature. The resulting solution was stirred at room temperature. t. for 16 hours. The reaction mixture was quenched using sat. aq. Na ₂ CO ₃ (200 ml), extracted with DCM (4x200 ml) and the combined organic layers were dried over MgSO ₄ , filtered and evaporated. The crude product was purified by fcc, eluting with a gradient of 0 to 20% EtOAc in petroleum ether to give the title compound (0.91 g, 10%) as a gray solid.

- 34 043669

1H NMR (400 MHz, CDCI3) 5.03 (2H, s), 8.22 (1H, s), 9.19 (1H, s), 9.56 (1H, s);

mass/charge MN ⁺ 258.

Intermediate 27: 5-chloro-7-methylquinazolin-6-amine.

Cl

Pd(Ph ₃ P) ₄ (384 mg, 0.33 mmol) was added in one portion to 7-bromo-5-chloroquinazoline-6-amine (860 mg, 3.33 mmol), trimethylboroxine (2506 mg, 9.98 mmol ) and K2CO3 (920 mg, 6.65 mmol) in THF (15 ml) at RT. The reaction mixture was stirred at 80°C for 48 h, then allowed to cool to RT and purified by fcc, gradient elution 0 to 30% EtOAc in petroleum ether to give the title compound (540 mg, 84%) as a pale yellow solid.

1H NMR (400 MHz, DMSO) 2.43 (3H, d), 6.03 (2H, s), 7.68 (1H, s), 9.02 (1H, s), 9.38 (1H, s);

mass/charge MN ⁺ 194.

Intermediate 28: 7-methylquinazoline-6-amine.

Pd/C 10% (500 mg, 4.70 mmol), 5-chloro-7-methylquinazolin-6-amine (500 mg, 2.58 mmol) and Et ₃ N (0.720 ml, 5.16 mmol) in MeOH (25 ml) was stirred under hydrogen pressure of 3 atm. at room temperature for 3 days. The reaction mixture was filtered through celite and concentrated in vacuo. The resulting crude product was purified by flash chromatography with a C18 flash column, eluting from 0 to 30% MeCN in water to give the title compound (80 mg, 19%) as a yellow solid.

1H NMR (400 MHz, DMSO) 2.32 (3H, d), 5.71 (2H, s), 6.95 (1H, s), 7.62 (1H, s), 8.86 (1H, s), 9.14 (1H, s);

mass/charge MN ⁺ 160.

Intermediate 29: Ethyl 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate.

CI^N^NH ό

Potassium carbonate (62.50 g, 452.41 mmol) was added to ethyl 2,4-dichloropyrimidine-5-carboxylate (40.00 g, 180.97 mmol) and tetrahydro-2H-pyran-4-amine hydrochloride (24 .90 g, 181.0 mmol) in acetonitrile (1 L). The reaction mixture was stirred at room temperature for 16 hours. The resulting precipitate was collected by filtration, washed with THF (750 ml) and the combined organic layers were concentrated in vacuo. The crude product was purified by fcc, eluting with a gradient of 0 to 2% THF in DCM to give the title compound (37.74 g, 73%) as a pale yellow solid.

1H NMR (400 MHz, DMSO) 1.32 (3H, t), 1.54-1.63 (2H, m), 1.85-1.89 (2H, m), 3.43-3.49 (2H, m), 3.83-3.88 (2H, m), 4.12-4.26 (1H, m), 4.29-4.34 (2H, m), 8.34 (1H , d), 8.64 (1H, s);

mass/charge MN ⁺ 286.

Intermediate 30: 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylic acid.

A solution of LiOH (13.11 g, 547.37 mmol) in water (800 ml) was added to a stirred solution of ethyl 2-chloro-4-((tetrahydro-2H-pyran-4-yl)amino)pyrimidine-5-carboxylate (78 .20 g, 273.7 mmol) in THF (800 ml). The reaction mixture was stirred at RT for 3 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting precipitate was collected by filtration, washed with water (500 ml) and dried in vacuo to give the title compound (66.4 g, 92%) as a white solid.

1H NMR (300 MHz, DMSO) 1.48-1.61 (2H, m), 1.85-1.89 (2H, m), 3.41-3.49 (2H, m), 3.81 -3.87 (2H, m), 4.10-4.22 (1H, m), 8.54 (1H, d), 8.59 (1H, s);

- 35 043669 mass/charge MN ⁺ 258.

Intermediate 31: 2-chloro-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8-one.

Triethylamine (25.4 g, 251.5 mmol) was added to 2-chloro-4-((tetrahydro-2H-pyran-4yl)amino)pyrimidine-5-carboxylic acid (64.8 g, 251.5 mmol) and diphenylphosphoryl azide (69.2 g, 251.5 mmol) in DMA (330 ml). The reaction mixture was stirred at room temperature for 1 hour, then stirred at 120°C for 16 hours. The reaction mixture was poured into ice (2 L), the precipitate was collected by filtration, washed with water (400 ml) and dried in vacuo with obtaining the title compound (44.8 g, 70%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.66-1.70 (2H, m), 2.39-2.47 (2H, m), 3.45 (2H, t), 3.95-3, 99 (2H, m), 4.38-4.46 (1H, m), 8.14 (1H, s), 11.65 (1H, s);

mass/charge MN ⁺ 255.

Intermediate 32: 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8Hpurin-8-one.

A solution of NaOH (31.0 g, 775.50 mmol) in water (80 ml) was added to a stirred solution of 2-chloro9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purine-8 -one (39.5 g, 155.1 mmol) and MeI (48.5 ml, 775.5 mmol) in THF (720 ml). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was partially concentrated in vacuo, then diluted with water. The resulting precipitate was collected by filtration, washed with water (300 ml) and dried in vacuo to give the title compound (32.5 g, 69%) as a white solid.

¹ H NMR (400 MHz, DMSO) 1.67-1.71 (2H, m), 2.39-2.48 (2H, m), 3.37 (3H, s), 3.46 (2H, t), 3.96-3.99 (2H, m), 4.42-4.50 (1H, m), 8.37 (1H, s);

mass/charge MN ⁺ 269.

Intermediate 33: Ethyl 2-chloro-4-(((1s,4s)-4-hydroxycyclohexyl)amino)pyrimidine 5-carboxylate.

Potassium carbonate (78.00 g, 565.5 mmol) was added to ethyl 2,4-dichloropyrimidine-5-carboxylate (50.00 g, 226.2 mmol) and (1s,4s)-4-aminocyclohexane-1- ol hydrochloride (34.30 g, 226.2 mmol) in acetonitrile (700 ml) at room temperature in an air atmosphere. The reaction mixture was stirred at room temperature for 16 hours, then filtered through a Celite® pad. The filtrate was partially concentrated in vacuo and the resulting precipitate was collected by filtration, washed with MeCN (100 mL) and dried in vacuo to give the title compound (41.0 g, 61%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.32 (3H, t), 1.42-1.58 (2H, m), 1.60-1.75 (6H, m), 3.66 (1H, d), 4.06 (1H, dd), 4.33 (2H, q), 4.57 (1H, d), 8.46 (1H, d), 8.63 (1H, s);

mass/charge MN ⁺ 300.

- 36 043669

Intermediate carboxylic acid.

connection 34:

2-chloro-4-((( 1 s,4s)-4-hydroxycyclohexyl)amino)pyrimidine-5 LiOH (9.75 g, 407.00

mmol) was added to ethyl 2-chloro-4-(((1s,4s)-4hydroxycyclohexyl)amino)pyrimidine-5-carboxylate (61.0 g, 203.50 mmol) in THF (400 ml) and water (400 ml) at room temperature in air atmosphere. The reaction mixture was stirred at room temperature for 16 hours, then partially concentrated in vacuo and acidified to pH~2 with 2 M aq. HCl solution. The resulting precipitate was collected by filtration, washed with water (500 ml) and dried in vacuo to give the title compound (52 g, 94%) as a white solid.

1H NMR (400 MHz, DMSO) 1.51 (2H, d), 1.58-1.75 (6H, m), 3.66 (1H, s), 4.00-4.07 (1H, m ), 4.56 (1H, s), 8.59 (1H, s), 8.69 (1H, d), 13.82 (1H, s);

mass/charge MH+ 272.

Intermediate 35: 2-chloro-9-((1s,4s)-4-hydroxycyclohexyl)-7,9-dihydro-8H-purin-8-one.

Triethylamine (28.2 ml, 202.4 mmol) was added to 2-chloro-4-(((1s,4s)-4hydroxycyclohexyl)amino)pyrimidine-5-carboxylic acid (55.0 g, 202.4 mmol) in acetonitrile (550 ml) at room temperature in an air atmosphere. The reaction mixture was stirred at room temperature for 15 minutes. DPPA (55.7 g, 202.4 mmol) was added and the reaction mixture was stirred at room temperature for 30 minutes and then heated at 90°C for 6 hours. The reaction mixture was allowed to cool to room temperature and poured into water (4 l). The precipitate was collected by filtration, washed with water (1 L) and dried in vacuo to give the title compound (34.9 g, 64.1%) as a white solid.

Mass/charge MN ⁺ 269.

Intermediate 36: 2-chloro-9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl-7,9-dihydro-8Hpurin-8-one.

Iodomethane (31.70 g, 223.30 mmol) was added to 2-chloro-9-((1s,4s)-4-hydroxycyclohexyl)-7,9dihydro-8H-purin-8-one (30.00 g, 111 .65 mmol), NaOH (22.33 g, 558.24 mmol) in THF (300 ml) and water (150 ml) at 25°C. The reaction mixture was stirred at 25°C for 16 hours. The reaction mixture was concentrated in vaccuo. The precipitate was collected by filtration, washed with water (250 ml) and dried in vacuo to give the title compound (24.02 g, 76%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.43-1.61 (4H, m), 1.79 (2H, d), 2.54-2.68 (2H, m), 3.34 (3H, s), 3.87 (1H, s), 4.15-4.21 (1H, m), 4.46 (1H, d), 8.34 (1H, s);

mass/charge MN ⁺ 283.

- 37 043669

Intermediate 37: Ethyl 2-chloro-4-(((1g,4g)-4-hydroxycyclohexyl)amino)pyrimidine 5-carboxylate.

DIPEA (35.10 g, 271.5 mmol) was added dropwise to ethyl 2,4-dichloropyrimidine-5-carboxylate (40 g, 181.0 mmol) and (1 g, 4 g)-4-aminocyclohexan-1-ol (20.84 g, 181.0 mmol) in acetonitrile (1.25 L) at 0°C. The reaction mixture was allowed to warm to room temperature, then stirred at room temperature for 16 hours. The resulting precipitate was collected by filtration, washed with THF (500 ml), and the combined organic layers were isolated and concentrated in vacuo. The resulting crude product was purified by fcc, eluting gradient from 0 to 3% THF in DCM to give the title compound (42.0 g, 77%) as a pale yellow solid.

¹ H NMR (300 MHz, DMSO) 1.23-1.45 (7H, m), 1.82-1.95 (4H, m), 3.47-3.48 (1H, m), 3, 86-3.95 (1H, m), 4.27-4.34 (2H, m), 4.63 (1H, d), 8.26 (1H, d), 8.60 (1H, s) ;

mass/charge MN ⁺ 300.

Intermediate 38: 2-chloro-4-(((1g,4g)-4-hydroxycyclohexyl)amino)pyrimidine-5-carboxylic acid.

A solution of LiOH (6.71 g, 280.23 mmol) in water (420 ml) was added to a stirred solution of ethyl 2-chloro-4-(((1g,4g)-4-hydroxycyclohexyl)amino)pyrimidine-5-carboxylate (42 .00 g, 140.1 mmol) in THF (420 ml). The reaction mixture was stirred at RT for 16 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting precipitate was collected by filtration, washed with water (350 ml) and dried in vacuo to give the title compound (34.29 g, 90%) as a white solid, which was used without further purification.

1H NMR (300 MHz, DMSO) 1.24-1.43 (4H, m), 1.84 (2H, d), 1.94 (2H, d), 3.44-3.50 (1H, m ), 3.88-3.90 (1H, m), 8.47 (1H, d), 8.58 (1H, s), 13.79 (1H, s), 1 exchanged proton was not observed;

mass/charge MN ⁺ 272.

Intermediate 39: 2-chloro-9-((1g,4g)-4-hydroxycyclohexyl)-7,9-dihydro-8H-purin-8-one.

Diphenylphosphoryl azide (38.3 g, 139.1 mmol) was added dropwise to 2-chloro-4-(((1g,4g)-4hydroxycyclohexyl)amino)pyrimidine-5-carboxylic acid (36.0 g, 132.5 mmol ) and added triethylamine (18.47 ml, 132.50 mmol) in THF (800 ml) at room temperature. The reaction mixture was stirred at 100°C for 12 hours, then allowed to cool to room temperature, concentrated in vacuo and the residue was diluted with water (700 ml). The solid was collected by filtration, dried in vacuo and triturated with DCM to give the title compound (18.36 g, 51%) as a white solid.

Mass/charge MN ⁺ 269.

Intermediate 40: 2-chloro-9-((1g,4g)-4-hydroxycyclohexyl)-7-methyl-7,9-dihydro-8Hpurin-8-one.

- 38 043669

Sodium hydroxide (26.0 g, 651.28 mmol) in water (350 ml) was added in one portion to 2-chloro-9((1g,4g)-4-hydroxycyclohexyl)-7,9-dihydro-8H-purine- 8-one (35.0 g, 130.3 mmol) and methyl iodide (40.7 ml, 651.3 mmol) in THF (700 ml) at room temperature. The reaction mixture was stirred at room temperature for one hour, then partially concentrated in vacuo and the resulting solid isolated by filtration and dried in vacuo to give the title compound (31.6 g, 86%) as a light yellow solid.

1H NMR (300 MHz, DMSO) 1.16-1.45 (2H, m), 1.61-1.81 (2H, m), 1.87-2.03 (2H, m), 2.15 -2.39 (2H, m), 3.35 (3H, s), 3.40-3.60 (1H, m), 4.00-4.28 (1H, m), 4.70 (1H , d), 8.35 (1H, s);

mass/charge MN ⁺ 283.

Intermediate 41: Ethyl (S)-2-chloro-4-((tetrahydro-2H-pyran-3-yl)amino)pyrimidine-5-carboxylate.

rr ^a CI^N^NH (S)-tetrahydro-2H-pyran-3-amine hydrochloride (1.99 g, 14.48 mmol) in MeCN (10 ml) was added dropwise to the DIPEA mixture (6.30 ml, 36.19 mmol) and ethyl 2,4-dichloropyrimidine-5-carboxylate (3.20 g, 14.48 mmol) in MeCN (60 ml) at 0° C. for a period of 5 minutes under air. The reaction mixture was stirred for 4 hours, allowed to warm slowly to room temperature, then stirred at room temperature for 18 hours and concentrated in vacuo, diluted with EtOAc (100 ml) and washed with water, then sat. saline solution. The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo and purified by fcc eluting with 0-40% EtOAc in n-heptane to give the title compound (3.24 g, 78%) as a yellow oil.

1H NMR (400 MHz, DMSO) 1.32 (3H, t), 1.49-1.6 (1H, m), 1.63-1.79 (2H, m), 1.83-1.94 (1H, m), 3.48 (1H, dd), 3.54 -3.65 (2H, m), 3.74 (1H, dd), 4.08-4.19 (1H, m), 4.33 (2H, q), 8.57 (1H, d), 8.64 (1H, s);

mass/charge [M-N] ^- 284.

Intermediate 42: 2-chloro-4-[[(3S)-tetrahydropyran-3-yl]amino]pyrimidine-5-carboxylic acid.

ABOUT

CI^N^NH

Lithium hydroxide hydrate (0.933 g, 22.23 mmol) was added in one portion to ethyl (S)-2-chloro-4((tetrahydro-2H-pyran-3-yl)amino)pyrimidin-5-carboxylate (3.241 g, 11.12 mmol) in THF (20 ml) and water (20 ml) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 16 hours. The reaction mixture was partially concentrated in vacuo, then acidified with 2 M aq. HCl solution. The resulting precipitate was collected by filtration, washed with water (50 ml) and air dried overnight. The resulting white solid was further dried in vacuo at 50°C for 24 hours to obtain the title compound (2.40 g, 84%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.55 (1H, dq), 1.61-1.77 (2H, m), 1.85-1.95 (1H, m), 3.45 (1H, dd), 3.59 (2H, t), 3.75 (1H, dd), 4.06-4.16 (1H, m), 8.60 (1H, s), 8.76 (1H, d ), 13.62 (1H, s);

mass/charge MN ⁺ 258.

Intermediate 43: (S)-2-chloro-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin-8-one.

Diphenylphosphoryl azide (2.00 ml, 9.29 mmol) was added in one portion to a solution of 2-chloro-4-[[(3S)tetrahydropyran-3-yl]amino]pyrimidine-5-carboxylic acid (2.40 g, 9. 29 mmol) and triethylamine (1.30 ml, 9.29 mmol) in THF (50 ml) at room temperature. The reaction mixture was stirred at 80°C for 24 hours. The reaction mixture was allowed to cool to room temperature, then poured into water (40 ml), then partially concentrated in vacuo, which led to the formation of a white precipitate, which was isolated by filtration, dried in vacuo, washed with water, air dried, then dried in vacuo at 50 ° C with obtaining the title compound (1.84 g, 78%) as a white solid.

1H NMR (400 MHz, DMSO) 1.61-1.82 (2H, m), 1.88-1.99 (1H, m), 2.4-2.49 (1H, m), 3.3 -3.37 (1H, m),

3.78-3.93 (3H, m), 4.2-4.32 (1H, m), 8.13 (1H, s), 11.63 (1H, s);

mass/charge MN ⁺ 255.

Intermediate 44: 2-chloro-7-methyl-9-[(3S)-tetrahydropyran-3-yl]purin-8-one.

Sodium hydride (60%) (0.434 g, 10.86 mmol) was added portionwise to (S)-2-chloro-9-(tetrahydro-2Hpyran-3-yl)-7,9-dihydro-8H-purine-8- onu (1.843 g, 7.24 mmol) in DMF (25 ml) at 0°C. The reaction mixture was stirred for 30 min, then iodomethane (1.36 mL, 21.71 mmol) was added dropwise. The reaction mixture was stirred at 0°C for 1 hour, then quenched with water (50 ml) and the resulting precipitate was filtered and dried in vacuo to obtain the title compound (1.62 g, 83%) as a cream solid.

^1H NMR (400 MHz, DMSO) 1.64-1.82 (2H, m), 1.9-1.98 (1H, m), 2.41-2.48 (1H, m), 3, 32-3.38 (4H, m), 3.79-3.91 (3H, m), 4.25-4.34 (1H, m), 8.35 (1H, s);

mass/charge MN ⁺ 269.

Intermediate 45: Ethyl 2-chloro-4-[[(3R)-tetrahydropyran-3-yl]amino]pyrimidine-5-carboxylate.

ABOUT

CI^N^NH (R)-tetrahydro-2H-pyran-3-amine hydrochloride (1.00 g, 7.27 mmol) in acetonitrile (5 ml) was added dropwise to the DIPEA mixture (3.16 ml, 18. 17 mmol) and ethyl 2,4-dichloropyrimidine-5-carboxylate (1.606 g, 7.27 mmol) in acetonitrile (30 ml) at 0° C. for a period of 5 minutes under air. The reaction mixture was stirred for 4 hours, allowed to warm slowly to room temperature and then stirred at room temperature overnight. The reaction mixture was concentrated in vacuo, diluted with EtOAc (100 ml) and washed with water, then sat. saline solution. The organic layer was isolated and dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in n-heptane to give the title compound (0.936 g, 45%) as a yellow oil.

^1H NMR (400 MHz, DMSO) 1.33 (3H, t), 1.57 (1H, dt), 1.71 (2H, dtd), 1.91 (1H, ddt), 3.48 (1H , dd), 3.55-3.66 (2H, m), 3.75 (1H, dd), 4.11-4.2 (1H, m), 4.33 (2H, q), 8, 58 (1H, d), 8.65 (1H, s);

mass/charge MN ⁺ 286.

Intermediate 46: 2-chloro-4-[[(3R)-tetrahydropyran-3-yl]amino]pyrimidine-5-carboxylic acid.

ABOUT

CI^N^NH

Lithium hydroxide hydrate (276 mg, 6.57 mmol) was added in one portion to ethyl 2-chloro-4-[[(3R)tetrahydropyran-3-yl]amino]pyrimidine-5-carboxylate (939 mg, 3.29 mmol ) in THF (1.23 ml) and water (4.10 ml) at RT. The reaction mixture was stirred at RT for 30 min, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting solid was isolated by filtration and dried at 45°C in vacuo overnight to give the title compound (806 mg, 95%) as a white solid.

1H NMR (400 MHz, DMSO) 1.56 (1H, dq), 1.70 (2H, ddt), 1.91 (1H, ddt), 3.46 (1H, dd), 3.60 (2H, t), 3.76 (1H, dd), 4.12 (1H, d), 8.61 (1H, s), 8.77 (1H, d), one exchanged proton was not observed;

mass/charge MN ⁺ 258.

- 40 043669

Intermediate 47: 2-chloro-9-[(3R)-tetrahydropyran-3-yl]-7H-purin-8-one.

Diphenylphosphoryl azide (0.674 ml, 3.13 mmol) was added in one portion to a solution of 2-chloro-4-[[(3R)tetrahydropyran-3-yl]amino]pyrimidine-5-carboxylic acid (806 mg, 3.13 mmol) and triethylamine (0.436 ml, 3.13 mmol) in THF (17.3 ml) at room temperature. The reaction mixture was heated at 80°C for 24 hours, then allowed to cool to room temperature and poured into water (20 ml ). The resulting mixture was partially concentrated in vacuo and the resulting precipitate was isolated by filtration, washed with water, air dried for 2 hours, then in vacuo overnight at 45°C to give the title compound (565 mg, 71%) as white solid matter.

^1H NMR (400 MHz, DMSO) 1.64-1.83 (2H, m), 1.93 (1H, d), 2.4-2.49 (1H, m), 3.35 (1H, dd), 3.8-3.92 (3H, m), 4.21-4.36 (1H, m), 8.13 (1H, s), 11.64 (1H, s);

mass/charge MH+ 255.

Intermediate 48: 2-chloro-7-methyl-9-[(3R)-tetrahydropyran-3-yl]purin-8-one.

Sodium hydride (60%) (133 mg, 3.33 mmol) was added portionwise to 2-chloro-9-[(3R)-tetrahydropyran3-yl]-7H-purin-8-one (565 mg, 2.22 mmol) in DMF (5.13 ml) at 0°C. The reaction mixture was stirred for 30 min, then iodomethane (0.42 mL, 6.66 mmol) was added dropwise. The reaction mixture was stirred at 0°C for 1 hour, then quenched with water (50 ml) and the resulting precipitate was isolated by filtration and dried in vacuo to give the title compound (535 mg, 90%) as a white solid.

1H NMR (400 MHz, DMSO) 1.73 (2H, dddd), 1.94 (1H, d), 2.41-2.49 (1H, m), 3.36 (4H, s), 3, 81-3.92 (3H, m), 4.24-4.36 (1H, m), 8.36 (1H, s);

mass/charge MH+ 269.

Intermediate 49: Ethyl 2-chloro-4-[(4-oxocyclohexyl)amino]pyrimidine 5-carboxylate.

DIPEA (4.19 ml, 24.0 mmol) was added dropwise to ethyl 2,4-dichloropyrimidine-5-carboxylate (4.42 g, 20.0 mmol) and 4-aminocyclohexan-1-one hydrochloride (2, 99 g, 20.0 mmol) in acetonitrile (100 ml) at 0°C for a period of 2 minutes. The reaction mixture was allowed to warm to RT, then stirred at RT for 16 h, then concentrated in vacuo and purified by fcc, eluting with a gradient of 0 to 5% EtOAc in DCM to give the title compound (3, 42 g, 57%) as a white solid.

^1H NMR (400 MHz, CDCl3) 1.41 (3H, t), 1.82-1.97 (2H, m), 2.28-2.41 (2H, m), 2.44-2, 62 (4H, m), 4.38 (2H, q), 4.52-4.66 (1H, m), 8.51-8.59 (1H, m), 8.71 (1H, s) ;

mass/charge MH+ 298.

- 41 043669

Intermediate 50: 2-chloro-4-((4-oxocyclohexyl)amino)pyrimidine-5-carboxylic acid.

LiOH (0.502 g, 20.96 mmol) was added in one portion to ethyl 2-chloro-4-[(4oxocyclohexyl)amino]pyrimidine-5-carboxylate (3.12 g, 10.48 mmol) in THF (25 ml) and water (25 ml) at 0°C. The reaction mixture was stirred at RT for 48 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting precipitate was isolated by filtration, washed with water (20 mL) and dried in vacuo to give the title compound (2.80 g, 99%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.80-1.98 (2H, m), 2.11-2.31 (4H, m), 2.50-2.63 (2H, m), 4, 38-4.52 (1H, m), 8.62 (2H, d), 13.81 (1H, s);

mass/charge MN ⁺ 270.

Intermediate 51: 2-chloro-9-(4-oxocyclohexyl)-7,9-dihydro-8H-purin-8-one.

Diphenylphosphoryl azide (4.00 mL, 18.54 mmol) was added in one portion to 2-chloro-4-((4oxocyclohexyl)amino)pyrimidine-5-carboxylic acid (5.00 g,

18.54 mmol) and Et ₃ N (2.58 ml,

18.54 mmol) in THF (80 ml) at room temperature. The reaction mixture was heated at 80°C for 16 hours, then allowed to cool to room temperature and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 40% EtOAc in DCM to give the title compound (3.50 g, 71%) as a white solid.

¹ H NMR (400 MHz, DMSO) 2.03-2.14 (2H, m), 2.25-2.35 (2H, m), 2.54-2.64 (2H, m), 2, 64-2.77 (2H, m), 4.72-4.85 (1H, m), 8.15 (1H, s), 11.65-11.71 (1H, m);

mass/charge MN ⁺ 267.

Intermediate 52: 2-chloro-7-methyl-9-(4-oxocyclohexyl)-7,9-dihydro-8H-purin-8-one.

NaH (0.525 g, 13.12 mmol) was added in one portion to 2-chloro-9-(4-oxocyclohexyl)-7,9-dihydro8H-purin-8-one (3.50 g, 13.12 mmol) in DMF (50 ml) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 30 minutes. MeI (2.462 mL, 39.37 mmol) was added and the reaction mixture was stirred at RT for 16 h, then poured into water (150 mL). The resulting precipitate was isolated by filtration, washed with water (50 ml) and dried in vacuo to give the title compound (3.30 g, 90%) as a white solid.

^1H NMR (400 MHz, DMSO) 2.03-2.14 (2H, m), 2.26-2.37 (2H, m), 2.53-2.65 (2H, m), 2, 65-2.77 (2H, m), 3.37 (3H, s), 4.76-4.89 (1H, m), 8.38 (1H, s);

mass/charge MN ⁺ 281.

Intermediate 53: 2-chloro-9-(4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro8H-pURin-8-one.

Bromomethylmagnesium (3 M, 0.89 ml, 2.67 mmol) was added to 2-chloro-7-methyl-9-(4-oxocyclohexyl)- 42 043669

7,9-dihydro-8H-purin-8-one (500 mg, 1.78 mmol) in THF (10 ml) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 4 hours, then concentrated in vacuo. The resulting crude product was purified by flash chromatography on a C18 column, eluting from 0 to 100% MeOH in water to give the title compound (400 mg, 76%) as a white solid (mixture of diastereoisomers).

1H NMR (main diastereoisomer) (300 MHz, CDCl ₃ ) 1.30 (3H, s), 1.47 (1H, s), 1.51-1.92 (6H, m), 2.44-2, 83 (2H, m), 3.44 (3H, s), 4.26-4.50 (1H, m), 8.01 (1H, s);

mass/charge MH+ 297.

Intermediate 54: Ethyl 2-chloro-4-(((1s,4s)-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate.

DIPEA (8.76 mL, 50.31 mmol) was added dropwise to a mixture of (1s,4s)-4-amino-1-methylcyclohexan-1ol (5.00 g, 38.70 mmol) and ethyl-2,4- dichloropyrimidine-5-carboxylate (8.55 g, 38.70 mmol) in acetonitrile (143 ml) at -5°C for a period of 15 minutes under air. The reaction mixture was stirred for 2 hours, then allowed to warm slowly to room temperature, concentrated in vacuo, diluted with EtOAc (200 ml) and washed with water, then sat. saline solution. The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude mixture was suspended in DCM (20 ml) and the resulting solid was isolated by filtration and washed with DCM (5 ml) to obtain the title compound (3.8 g). The filtrate was purified by fcc, eluting with a gradient of 0 to 70% EtOAc in n-heptane to give additional title compound (5.3 g). Both batches were combined to give the title compound (9.10 g, 75%) as a white solid.

1H NMR (400 MHz, DMSO) 1.13 (3H, s), 1.32 (3H, t), 1.43 (2H, td), 1.53-1.61 (2H, m), 1, 69 (4H, tt), 3.85-3.99 (1H, m), 4.15 (1H, s), 4.32 (2H, q), 8.27 (1H, d), 8.62 (1H, s);

mass/charge MH+ 314.

Intermediate 55: 2-chloro-4-(((1s,4s)-4-hydroxy-4methylcyclohexyl)amino)pyrimidine-5-carboxylic acid.

CI^N^NH

Lithium hydroxide hydrate (2.17 g, 51.63 mmol) was added in one portion to ethyl 2-chloro-4-(((1s,4s)-4hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate (8 .10 g, 25.81 mmol) in THF (97 ml) and water (32.3 ml) at RT. The reaction mixture was stirred at RT for 3 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution, and the resulting solid was isolated by filtration to obtain the title compound (7.35 g, 100%) as a white solid.

1H NMR (400 MHz, DMSO) 1.13 (3H, s), 1.43 (2H, td), 1.52-1.75 (6H, m), 3.89 (1H, qd), 4, 15 (1H, s), 8.50 (1H, d), 8.58 (1H, s), 13.75 (1H, s);

mass/charge MH+ 286.

Intermediate 56: 2-chloro-9-((1s,4s)-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H purin-8-one.

Diphenylphosphoryl azide (4.79 mL, 22.22 mmol) was added in one portion to a solution of 2-chloro-4(((1s,4s)-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (6.35 g, 22.22 mmol) and triethylamine (3.10 ml, 22.22 mmol) in THF (123 ml) at room temperature. The reaction mixture was heated at 80°C in

- 43 043669 for 24 hours, then allowed to cool to room temperature and poured into water (100 ml). The resulting mixture was partially concentrated in vacuo and the resulting precipitate was isolated by filtration, washed with water, then dried in vacuo at 45°C to give the title compound (5.39 g, 86%) as a white solid.

Ή NMR (400 MHz, DMSO) 1.15 (3H, s), 1.39-1.52 (4H, m), 1.66 (2H, d), 2.54-2.71 (2H, m ), 4.10 (2H, qd), 8.11 (1H, s), 11.55 (1H, s);

mass/charge MH+ 283.

Intermediate 57: 2-chloro-9-((1s,4s)-4-hydroxy-4-methylcyclohexul)-7-methyl-7,9dihydro-8H-purin-8-one.

M aq. sodium hydroxide solution (37.5 mL, 74.98 mmol) was added in one portion to 2-chloro-9((1s,4s)-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purine-8 -one (4.24 g, 15.00 mmol) and iodomethane (4.69 ml, 74.98 mmol) in THF (73.2 ml) at room temperature in air. The reaction mixture was stirred at room temperature for 3 hours, then partially concentrated in vacuo. The resulting white precipitate was isolated by filtration, washed with water and dried in vacuo at 45°C to obtain the title compound (3.64 g, 82%) as a white solid.

1H NMR (400 MHz, DMSO) 1.15 (3H, s), 1.47 (4H, d), 1.66 (2H, d), 2.58-2.65 (2H, m), 3, 36 (3H, s), 4.1-4.19 (2H, m), 8.33 (1H, s);

mass/charge MH+ 297.

Intermediate 58: Ethyl 2-chloro-4-(((1t,4t)-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate.

CI^N^NH

BUT ^g

DIPEA (17.53 mL, 100.62 mmol) was added dropwise to a mixture of (1t,4t)-4-amino-1-methylcyclohexan1-ol (10.00 g, 77.40 mmol) and ethyl-2,4- dichloropyrimidine-5-carboxylate (17.11 g, 77.40 mmol) in acetonitrile (300 ml) at -5°C for 5 min in air. The reaction mixture was stirred for 18 hours, allowed to warm slowly to room temperature, then concentrated in vacuo, diluted with EtOAc (200 ml) and washed with water, then sat. saline solution. The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in n-heptane to give the title compound (17.85 g, 74%) as a white solid.

1H NMR (400 MHz, DMSO) 1.16 (3H, s), 1.32 (3H, t), 1.46-1.58 (6H, m), 1.82-1.94 (2H, m ), 4.06 (1H, dq), 4.26 (1H, s), 4.32 (2H, q), 8.45 (1H, d), 8.61 (1H, s);

mass/charge MH+ 314.

Intermediate 59: 2-chloro-4-(((1t,4t)-4-hydroxy-4methylcyclohexyl)amino)pyrimidine-5-carboxylic acid.

n^ ^CO2H

CI^N^NH

Lithium hydroxide hydrate (4.77 g, 113.77 mmol) was added in one portion to ethyl 2-chloro-4-(((1t,4t)4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylate ( 17.85 g, 56.89 mmol) in THF (213 ml) and water (71.1 ml) at RT. The reaction mixture was stirred at RT for 30 min, then partially concentrated in vacuo and acidified with using 2 M aq. HCl solution. The resulting precipitate was isolated by filtration to give the title compound (14.78 g, 91%) as a white solid.

1H NMR (400 MHz, DMSO) 1.16 (3H, s), 1.43-1.56 (6H, m), 1.89 (2H, dt), 3.96-4.12 (1H, m ), 4.26 (1H,

- 44 043669

s), 8.58 (1H, s), 8.69 (1H, d), 13.73 (1H, s);

mass/charge MN ⁺ 286.

Intermediate 60: 2-chloro-9-((1g,4g)-4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8Hpurin-8-one.

Η ci^n^n / yu Η

Diphenylphosphoryl azide (11.15 mL, 51.73 mmol) was added in one portion to a solution of 2-chloro-4(((1r,4r)-4-hydroxy-4-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (14.78 g, 51.73 mmol) and triethylamine (7.21 ml, 51.73 mmol) in THF (286 ml) at room temperature. The reaction mixture was stirred at 80°C for 24 hours, then allowed to cool to room temperature ., then poured into water (200 ml). The resulting mixture was partially concentrated in vacuo. The resulting precipitate was isolated by filtration, washed with water and dried in vacuo at 45°C to obtain the title compound (12.53 g, 86%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.27 (3H, s), 1.53 (2H, td), 1.6-1.72 (4H, m), 2.24-2.44 (2H, m), 4.15 (1H, tt), 4.41 (1H, s), 8.12 (1H, s), 11.60 (1H, s);

mass/charge MN ⁺ 283.

Intermediate 61: 2-chloro-9-((1g,4g)-4-hydroxy-4-methylcyclohexyl)-7-methyl-7,9dihydro-8H-purin-8-one.

/n X >°ci^n^n /'OH

M sodium hydroxide solution (44.8 mL, 89.66 mmol) was added in one portion to 2-chloro-9-((1g,4g)4-hydroxy-4-methylcyclohexyl)-7,9-dihydro-8H-purine- 8-one (5.07 g, 17.93 mmol) and iodomethane (5.61 ml, 89.66 mmol) in THF (88 ml) at room temperature in air. The reaction mixture was stirred at room temperature for 5 hours, then partially concentrated in vacuo. The resulting solid was isolated by filtration, washed with water and dried in vacuo at 45°C to obtain the title compound (4.00 g, 75%) as a white solid.

1H NMR (400 MHz, DMSO) 1.27 (3H, s), 1.46-1.6 (2H, m), 1.67 (4H, d), 2.33 (2H, ddd), 3, 36 (3H, s), 4.19 (1H, ddt), 4.45 (1H, s), 8.35 (1H, s);

mass/charge MN ⁺ 297.

Intermediate 62: Ethyl 2-chloro-4-((4-hydroxy-1methylcyclohexyl)amino)pyrimidine-5-carboxylate.

DIPEA (4.28 mL, 24.49 mmol) was added dropwise to ethyl 2,4-dichloropyrimidine-5-carboxylate (2.46 g, 11.13 mmol) and 4-amino-4-methyl-cyclohexanol hydrochloride ( 2.00 g, 11.13 mmol) in acetonitrile (40 ml) at 0°C for 5 min. The reaction mixture was allowed to warm to room temperature, then stirred at room temperature for 6 hours and concentrated in vacuo, diluted with EtOAc (300 ml) and washed with sat. saline solution (100 mlx2). The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 20% EtOAc in n-heptane to give the title compound (2.82 g, 81%) as a pale yellow gum.

1H NMR (400 MHz, DMSO) 1.36-1.44 (3H, m), 1.44-1.58 (6H, m), 1.57-1.71 (1H, m), 1.72 -2.13 (3H, m), 2.41-2.54 (2H, m), 3.63-3.75 (1H, m), 4.30-4.42 (2H, m), 8 .52-8.59 (1H, m), 8.67 (1H, d);

mass/charge MN ⁺ 314.

- 45 043669

Intermediate 63: 2-chloro-4-((4-hydroxy-1-methylcyclohexyl)amino)pyrimidine-5-carboxylic acid.

LiOH (0.43 g, 17.97 mmol) was added in one portion to ethyl 2-chloro-4-((4-hydroxy-1methylcyclohexyl)amino)pyrimidine-5-carboxylate (2.82 g, 8.99 mmol) in THF (25 ml) and water (25 ml) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 5 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting precipitate was isolated by filtration, washed with water (20 mL) and dried in vacuo to give the title compound (2.17 g, 85%) as a white solid.

¹ H NMR (400 MHz, DMSO) 1.18-1.32 (2H, m), 1.34-1.52 (5H, m), 1.52-1.79 (2H, m), 2, 21-2.30 (2H, m), 3.37-3.49 (1H, m), 4.55 (1H, s), 8.59 (1H, d), 8.74 (1H, s) , 13.85 (1H, s);

mass/charge MH+286.

Intermediate 64: 2-chloro-9-(4-hydroxy-1-methylcyclohexyl)-7,9-dihydro-8H-purin8-one.

Diphenylphosphoryl azide (1.64 ml, 7.59 mmol) was added in one portion to 2-chloro-4-((4-hydroxy-1methylcyclohexyl)amino)pyrimidine-5-carboxylic acid (2.17 g, 7.59 mmol) and Et ₃ N (1.06 ml,

7.59 mmol) in THF (20 ml) at RT. The reaction mixture was heated at 80°C for 2 days, then concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in DCM to give the title compound (1.79 g, 83%) as a white solid.

L NMR (400 MHz, DMSO) 1.09-1.25 (2H, m), 1.34-1.64 (5H, m), 1.65-1.77 (2H, m), 3.17 (2H, d), 3.41-3.57 (1H, m), 4.07-4.15 (1H, m), 8.10 (1H, d), 11.61 (1H, s);

mass/charge MN ⁺ 283.

Intermediates 65 and 66: 2-chloro-9-((1s,4s)-4-hydroxy-1-methylcyclohexyl)-7-methyl-7,9dihydro-8H-purin-8-one and 2-chloro-9- ((1r,4r)-4-hydroxy-1-methylcyclohexyl)-7-methyl-7,9-dihydro-8Hpurin-8-one.

A solution of NaOH (1.27 g, 31.66 mmol) in water (24 ml) was added to the stirred mixture of 2-chloro-9-(4hydroxy-1-methylcyclohexyl)-7,9-dihydro-8H-purin-8-one (1.79 g, 6.33 mmol), iodomethane (1.97 ml, 31.66 mmol) and tetrabutylammonium bromide (0.204 g, 0.63 mmol) in DCM (40 ml) at room temperature. The reaction mixture was stirred at room temperature for 16 h, then extracted with DCM (3x50 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 40% EtOAc in DCM to give the title compounds. Minor product 2-chloro-9-((1s,4s)-4-hydroxy-1methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (0.26 g, 14%) as white solid.

^1H NMR (400 MHz, CDCl ₃ ) 1.66 (3H, s), 1.67-1.85 (4H, m), 2.19-2.31 (2H, m), 2.91-3 .02 (2H, m), 3.41 (3H, s), 3.89-3.99 (1H, m), 7.99 (1H, s), one exchange proton missing;

mass/charge MN ⁺ 297.

The main product is 2-chloro-9-((1g,4g)-4-hydroxy-1-methylcyclohexyl)-7-methyl-7,9-dihydro-8Hpurin-8-one (1.440 g, 77%) as a white solid substances.

^1H NMR (400 MHz, CDCl ₃ ) 1.42-1.50 (2H, m), 1.51 (3H, s), 1.58-1.88 (2H, m), 1.88-2 .00 (2H, m), 3.40 (3H, s), 3.52-3.63 (2H, m), 3.72-3.84 (1H, m), 7.99 (1H, s ), one exchange proton is missing;

mass/charge MN ⁺ 297.

- 46 043669

Intermediate 67: 2-chloro-7-methyl-9-(1-methyl-4-oxocyclohexyl)-7,9-dihydro-8Hpurin-8-one.

Dess-Martin periodinane (1.07 g, 2.53 mmol) was added to 2-chloro-9-((1g,4g)-4-hydroxy-1methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purine -8-one (0.50 g, 1.68 mmol) in DCM (10 ml). The reaction mixture was stirred at room temperature for 4 hours, then quenched with sat. aq. NaHCO ₃ solution (20 ml) and extracted with DCM (3x20 ml). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by flash chromatography with a C18 flash column, eluting gradient from 0 to 50% MeOH in water to give the title compound (0.43 g, 87%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.52 (3H, s), 1.88-2.05 (2H, m), 2.11-2.25 (2H, m), 2.37-2, 49 (2H, m), 3.34 (3H, s), 3.45-3.60 (2H, m), 8.37 (1H, s); mass/charge MN ⁺ 295.

Intermediate 68:

Dihydro-8H-purin-8-one.

2-chloro-9-((1s,4s)-4-hydroxy-1-methylcyclohexyl)-7-methyl-7,9-

Sodium borohydride (135 mg, 3.56 mmol) was added to 2-chloro-7-methyl-9-(1-methyl-4-oxocyclohexyl)7,9-dihydro-8H-purin-8-one (700 mg, 2 .37 mmol) in MeOH (15 ml) at room temperature. The reaction mixture was stirred for 2 hours at room temperature, then quenched with sat. aq. NH4Cl solution (1 ml) and concentrated in vacuo. The resulting crude mixture was purified by fcc, eluting gradient from 0 to 40% DCM in EtOAc to give the title compound (50 mg, 7%) as a white solid.

1H NMR (400 MHz, DMSO) 1.43-1.53 (5H, m), 1.54-1.65 (2H, m), 2.01-2.13 (2H, m), 2.79 -2.84 (2H, m), 3.32 (3H, s), 3.59-3.70 (1H, m), 4.51 (1H, d), 8.33 (1H, s);

mass/charge MN ⁺ 297.

Intermediate 69: Ethyl 2-chloro-4-[[(3R)-tetrahydrofuran-3-yl]amino]pyrimidine-5-carboxylate.

O rf ⁰ CI^N^NH <r

DIPEA (4.74 ml, 27.14 mmol) was added to ethyl 2,4-dichloropyrimidine-5-carboxylate (5.00 g, 22.62 mmol) and (R)-tetrahydrofuran-3-amine (1.97 g, 22.62 mmol) in MeCN (100 ml) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 4 hours, then concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 9% EtOAc in petroleum ether to give the title compound (4.90 g, 80%) as a yellow solid.

1H NMR (400 MHz, CDCl ₃ ) 1.40 (3H, t), 1.80-1.98 (1H, m), 2.29-2.46 (1H, m), 3.69-3, 78 (1H, m), 3.79-3.93 (1H, m), 3.95-4.05 (2H, m), 4.36 (2H, q), 4.68-4.90 ( 1H, m), 8.55 (1H, s), 8.66-8.71 (1H, m);

mass/charge MN ⁺ 272.

Intermediate 70: 2-chloro-4-[[(3R)-tetrahydrofuran-3-yl]amino]pyrimidine-5-carboxylic acid.

- 47 043669

A solution of LiOH (0.864 g, 36.07 mmol) in water (40.0 mL) was added to a stirred solution of ethyl 2-chloro-4-[[(3R)-tetrahydrofuran-3-yl]amino]pyrimidine-5-carboxylate (4 .90 g, 18.03 mmol) in THF (40 ml) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 3 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting precipitate was isolated by filtration, washed with water (20 mL) and dried in vacuo to give the title compound (3.90 g, 89%) as a white solid.

1H NMR (400 MHz, CDCl ₃ ) 2.01-2.14 (1H, m), 2.40-2.54 (1H, m), 3.89-4.23 (4H, m), 5, 01-5.13 (1H, m), 8.78 (1H, s), 9.08 (1H, d), one exchange proton was not observed;

mass/charge MN ⁺ 244.

Intermediate 71: 2-chloro-9-[(3R)-tetrahydrofuran-3-yl]-7H-purin-8-one.

Diphenylphosphoryl azide (3.45 ml, 16.01 mmol) was added to 2-chloro-4-[[(3R)-tetrahydrofuran-3yl]amino]pyrimidine-5-carboxylic acid (3.90 g, 16.01 mmol) and triethylamine (2.23 ml, 16.01 mmol) in THF (70 ml). The reaction mixture was heated at 80°C for 24 hours and then concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in DCM to give the title compound (3.20 g, 83%) as a white solid.

1H NMR (400 MHz, DMSO) 2.16-2.30 (1H, m), 2.35-2.48 (1H, m), 3.81-3.94 (2H, m), 3.94 -4.02 (1H, m), 4.05-4.15 (1H, m), 4.91-5.03 (1H, m), 8.14 (1H, s), 11.68 (1H , s);

mass/charge MN ⁺ 241.

Intermediate 72: 2-chloro-7-methyl-9-[(3R)-tetrahydro-3-furanyl]-7,9-dihydro-8Hpurin-8-one.

gАо ci^n^ ⁿ _u

ΟΑ

NaH (0.532 g, 13.30 mmol) was added to 2-chloro-9-[(3R)-tetrahydrofuran-3-yl]-7H-purin-8-one (3.2 g, 13.30 mmol) in DMF (40 ml) at 0°C. The reaction mixture was stirred at 0°C for 1 hour, then MeI (5.66 g, 39.89 mmol) was added and the reaction mixture was stirred at room temperature for 5 hours, then concentrated in vacuo. The crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in DCM to give the title compound (3.20 g, 94%) as a yellow solid.

1H NMR (300 MHz, MeOD) 2.28-2.47 (1H, m), 2.50-2.67 (1H, m), 3.46 (3H, s), 3.94-4.15 (3H, m), 4.23-4.37 (1H, m), 5.08-5.24 (1H, m), 8.23 (1H, s);

mass/charge MN ⁺ 255.

Intermediate 73: Ethyl 2-chloro-4-[[(3S)-tetrahydrofuran-3-yl]amino]pyrimidine-5-carboxylate.

DIPEA (4.74 ml, 27.14 mmol) was added dropwise to ethyl 2,4-dichloropyrimidine-5-carboxylate (5 g, 22.62 mmol) and (S)-tetrahydrofuran-3-amine (1.97 g, 22.62 mmol) in acetonitrile (100 ml) at 0°C for a period of 2 minutes. The reaction mixture was allowed to warm to room temperature, then stirred at room temperature for 16 hours and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 5% EtOAc in petroleum ether to give the title compound (4.60 g, 75%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.32 (3H, t), 1.83-1.95 (1H, m), 2.21-2.35 (1H, m), 3.61-3, 69 (1H, m), 3.69-3.92 (3H, m), 4.27-4.37 (2H, m), 4.57-4.68 (1H, m), 8.44 ( 1H, d), 8.63 (1H, s);

mass/charge MN ⁺ 272.

- 48 043669

Intermediate 74: 2-chloro-4-[[(3S)-tetrahydrofuran-3-yl]amino]pyrimidine-5-carboxylic acid.

LiOH (0.811 g, 33.86 mmol) was added in one portion to ethyl 2-chloro-4-[[(3S)-tetrahydrofuran-3yl]amino]pyrimidine-5-carboxylate (4.60 g, 16.93 mmol) in THF (50 ml) and water (25 ml) at 0°C. The reaction mixture was allowed to warm to room temperature, stirred at room temperature for 2 hours, partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting precipitate was isolated by filtration, washed with water (20 ml) and dried in vacuo to give the title compound (3.50 g, 85%) as a white solid.

1H NMR (400 MHz, DMSO) 1.81-1.93 (1H, m), 2.21-2.35 (1H, m), 3.60-3.68 (1H, m), 3.69 -3.94 (3H, m), 4.56-4.68 (1H, m), 8.63 (2H, d), 13.84 (1H, s);

mass/charge MN ⁺ 244.

Intermediate 75: 2-chloro-9-[(3S)-tetrahydro-3-furanyl]-7,9-dihydro-8H-purin-8-one.

Diphenylphosphoryl azide (3.10 mL, 14.37 mmol) was added in one portion to 2-chloro-4-[[(3S)tetrahydrofuran-3-yl]amino]pyrimidine-5-carboxylic acid (3.5 g, 14.4 mmol) and Et ₃ N (2.00 ml, 14.4 mmol) in THF (100 ml) at RT. The reaction mixture was heated at 80°C for 2 days. The solvent was removed under reduced pressure. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in petroleum ether to give the title compound (3.20 g, 93%) as a white solid.

^1H NMR (400 MHz, DMSO) 2.16-2.32 (1H, m), 2.35-2.48 (1H, m), 3.81-3.92 (2H, m), 3, 97 (1H, t), 4.10 (1H, q), 4.91-5.03 (1H, m), 8.14 (1H, s), 11.66 (1H, s);

mass/charge MN ⁺ 241.

Intermediate 76: 2-chloro-7-methyl-9-[(3S)-tetrahydro-3-furanyl]-7,9-dihydro-8Hpurin-8-one.

NaH (0.532 g, 13.30 mmol) was added in one portion to 2-chloro-9-[(3S)-tetrahydro-3-furanyl]-7,9dihydro-8I-purin-8-one (3.2 g, 13 .30 mmol) in DMF (30 ml) at 0°C. The reaction mixture was stirred at room temperature for 30 minutes. MeI (2.49 mL, 39.9 mmol) was added. The reaction mixture was stirred at room temperature for 16 hours, then quenched with water (5 ml) and concentrated in vacuo. The crude product was purified by fcc, eluting with a gradient of 0 to 40% EtOAc in petroleum ether to give the title compound (2.90 g, 86%) as a yellow solid.

1H NMR (400 MHz, DMSO) 2.18-2.32 (1H, m), 2.35-2.48 (1H, m), 3.36 (3H, s), 3.82-3.94 (2H, m), 3.98 (1H, t), 4.11 (1H, q), 4.95-5.07 (1H, m), 8.36 (1H, s);

mass/charge MN ⁺ 255.

Intermediate 77: Ethyl 2-chloro-4-[(1,1-dioxothian-4-yl)amino]pyrimidine-5-carboxylate.

DIPEA (7.68 ml, 44.0 mmol) was added dropwise to ethyl 2,4-dichloropyrimidine 5-carboxylate

- 49 043669 (4.42 g, 20.0 mmol) and 4-aminotetrahydro-2H-thiopyran-1,1-dioxide hydrochloride (3.71 g, 20.0 mmol) in acetonitrile (80 ml) at 0°C for a period of 5 minutes. The resulting mixture was stirred at RT for 6 hours, then concentrated in vacuo, diluted with EtOAc (500 ml) and washed with sat. saline solution (100 mlx2). The organic layer was isolated and dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 6% EtOAc in DCM to give the title compound (2.40 g, 36%) as a white solid.

1H NMR (400 MHz, CDCls) 1.42 (3H, t), 2.23-2.38 (2H, m), 2.40-2.51 (2H, m), 3.10-3.27 (4H, m), 4.34-4.50 (3H, m), 8.57 (1H, d), 8.73 (1H, s);

mass/charge MN ⁺ 334.

Intermediate 78: 2-chloro-4-[(1,1-dioxothian-4-yl)amino]pyrimidine-5-carboxylic acid.

LiOH (0.344 g,

14.38 mmol) was added in one portion to ethyl 2-chloro-4-[(1,1-dioxothian-4yl)amino]pyrimidine-5-carboxylate (2.40 g, 7.19 mmol) in THF (25 ml) and water (25 ml) at 0°C. The reaction mixture was stirred at RT for 5 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting precipitate was collected by filtration, washed with water (50 ml) and dried in vacuo to give the title compound (2.08 g, 95%) as a white solid.

^1H NMR (400 MHz, DMSO) 2.00-2.15 (2H, m), 2.18-2.30 (2H, m), 3.02-3.14 (2H, m), 3, 34-3.55 (2H, m), 4.27-4.42 (1H, m), 8.57 (1H, d), 8.61 (1H, s), 13.84 (1H, s) ;

mass/charge MN ⁺ 306.

Intermediate 79: 2-chloro-9-(1,1-dioxothian-4-yl)-7H-purin-8-one.

Diphenylphosphoryl azide (1.46 ml, 6.80 mmol) was added in one portion to 2-chloro-4-[(1,1dioxotian-4-yl)amino]pyrimidine-5-carboxylic acid (2.08 g, 6.80 mmol ) and triethylamine (0.948 ml, 6.80 mmol) in THF (40 ml) at RT. The reaction mixture was stirred at 80°C for 2 days, then poured into water (75 ml). The resulting precipitate was collected by filtration, washed with water (25 mL) and dried in vacuo to give the title compound (1.72 g, 84%) as a white solid.

1H NMR (400 MHz, DMSO) 2.05-2.15 (2H, m), 2.81-3.03 (2H, m), 3.07-3.23 (2H, m), 3.43 -3.56 (2H, m), 4.59-4.72 (1H, m), 8.15 (1H, s), 11.69 (1H, s);

mass/charge MH+303.

Intermediate 80: 2-chloro-9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-7,9dihydro-8I-purin-8-one.

NaH (0.337 g, 8.42 mmol) was added in one portion to 2-chloro-9-(1,1-dioxothian-4-yl)-7H-purin-8one (1.70 g, 5.62 mmol) in DMF (25 ml) at room temperature and the reaction mixture was stirred at room temperature for 30 minutes. MeI (0.53 ml, 8.42 mmol) was added and the reaction mixture was stirred at room temperature for 2 hours, then diluted with water (50 ml). The resulting precipitate was collected by filtration, washed with water (20 mL) and dried in vacuo to give the title compound (1.67 g, 94%) as a white solid.

1H NMR (400 MHz, DMSO) 2.05-2.15 (2H, m), 2.81-2.96 (2H, m), 3.09-3.21 (2H, m), 3.36 (3Н, s),

3.44-3.57 (2H, m), 4.64-4.77 (1H, m), 8.38 (1H, s);

mass/charge MN ⁺ 317.

Intermediate 81: 2-chloro-5-nitro-N-(oxetan-3-yl)pyrimidin-4-amine.

Oxetan-3-amine (1.507 g, 20.62 mmol) was added dropwise to 2,4-dichloro-5-nitropyrimidine (4.00 g, 20.62 mmol) and DIPEA (4.67 mL, 26.81 mmol ) in DCM (100 ml) at -78°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 2 hours, then washed successively with water (100 ml) and sat. saline solution (100 ml). The organic layer was filtered through phase separation filter paper and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 100% EtOAc in n-heptane to give the title compound (3.70 g, 78%) as a white solid.

1H NMR (400 MHz, DMSO) 4.71 (2H, t), 4.77 (2H, t), 5.09 (1H, qd), 9.06 (1H, s), 9.34 (1H, d).

Intermediate 82: 2-chloro-N4-(oxetan-3-yl)pyrimidin-4,5-diamine.

Platinum (10% on carbon) (0.313 g, 1.60 mmol) was added to 2-chloro-5-nitro-N-(oxetan-3yl)pyrimidin-4-amine (3.70 g, 16.04 mmol) in EtOAc (50 ml) at RT. The reaction mixture was purged with hydrogen and stirred under a hydrogen atmosphere at RT for 18 hours. The reaction mixture was diluted with MeOH (to solubilize the product) and filtered, washing the precipitate with MeOH. The combined MeOH layers were concentrated in vacuo to give the title compound (3.10 g, 96%) as an off-white solid.

^1H NMR (400 MHz, DMSO) 4.49 (2H, t), 4.83 (2H, t), 4.9-5.06 (3H, m), 7.45 (1H, s), 7 .50 (1H, d);

mass/charge MN ⁺ 201.

Intermediate 83: 2-chloro-9-(oxetan-3-yl)-7,9-dihydro-8H-purin-8-one.

2-chloro-N4-(oxetan-3-yl)pyrimidin-4,5-diamine (3.10 g, 15.45 mmol) was placed in a flask in THF (100 ml) at room temperature. Di(1H- imidazol-1-yl)methanone (4.01 g, 24.72 mmol) and the reaction mixture was refluxed for 1 hour, then allowed to cool to room temperature and partially concentrated (~50%) in vacuo . The resulting precipitate was isolated by filtration and dried in vacuo to give the title compound (2.75 g, 79%) as a white solid.

Ή NMR (400 MHz, DMSO) 4.77 (2H, dd), 5.24 (2H, t), 5.46 (1H, p), 8.15 (1H, s), 11.68 (1H, s); mass/charge MN ⁺ 227.

Intermediate 84: 2-chloro-7-methyl-9-(oxetan-3-yl)-7,9-dihydro-8H-purin-8-one.

Sodium hydride (60%) (0.73 g, 18.20 mmol) was added portionwise to 2-chloro-9-(oxetan-3-yl)-7,9dihydro-8H-purin-8-one (2.75 g , 12.13 mmol) in DMF (25 ml) at room temperature. The reaction mixture was stirred for 30 min, then cooled to 0°C and iodomethane (2.28 ml, 36.40 mmol) was added dropwise. The resulting solution was stirred at RT for 1 hour. The reaction mixture was poured into water, the solid was filtered and dried to give the title compound (2.80 g, 96%) as a cream solid.

1H NMR (400 MHz, DMSO) 3.36 (3H, s), 4.79 (2H, dd), 5.23 (2H, t), 5.50 (1H, p), 8.38 (1H, s);

- 51 043669 mass/charge MN ⁺ 241.

Intermediate 85: Ethyl 4-[(1-tert-butoxycarbonyl-4-piperidyl)amino]-2chloropyrimidine-5-carboxylate.

DIPEA (20.49 mL, 117.63 mmol) was added dropwise to a mixture of tert-butyl 4-aminopiperidine-1-carboxylate (18.12 g, 90.48 mmol) and ethyl 2,4-dichloropyrimidine-5-carboxylate ( 20.00 g, 90.48 mmol) in acetonitrile (334 ml) at -5°C for a period of 15 minutes under air. The reaction mixture was stirred for 2 hours, allowed to warm slowly to room temperature, then concentrated in vacuo, diluted with EtOAc (300 ml) and washed with water, then sat. saline solution. The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 40% EtOAc in n-heptane to give the title compound (24.56 g, 71%) as a pale yellow solid.

^1H NMR (400 MHz, DMSO) 1.32 (3H, t), 1.41 (9H, s), 1.43-1.53 (2H, m), 1.84-1.91 (2H, m), 2.9-3.03 (2H, m), 3.87 (2H, d), 4.17 (1H, ddt), 4.32 (2H, q), 8.31 (1H, d ), 8.64 (1H, s);

mass/charge MN ⁺ 385.

Intermediate 86: 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2chloropyrimidine-5-carboxylic acid.

Lithium hydroxide hydrate (5.36 g, 127.63 mmol) was added in one portion to ethyl 4-[(1-tert-butoxycarbonyl-4-piperidyl)amino]-2-chloropyrimidine-5-carboxylate (24.56 g, 63. 82 mmol) in THF (239 ml) and water (80 ml) at 20°C. The resulting solution was stirred at 25°C for 3 hours. The organic solvent was removed under reduced pressure. The resulting mixture was acidified with 2 M HCl. The resulting white solid was filtered to obtain the title compound (22.72 g, 100%) as a white solid.

1H NMR (400 MHz, DMSO) 1.37-1.51 (11H, m), 1.89 (2H, dd), 2.97 (2H, s), 3.86 (2H, d), 4, 14 (1H, qd), 8.56 (1H, d), 8.60 (1H, s);

mass/charge MN ⁺ 357.

Intermediate 87: tert-butyl 4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9yl)piperidin-1-carboxylate.

Diphenylphosphoryl azide (13.72 mL, 63.68 mmol) was added in one portion to a solution of 4-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid (22.72 g, 63 .68 mmol) and triethylamine (8.88 ml, 63.68 mmol) in THF (352 ml) at room temperature. The reaction mixture was heated at 80°C for 24 hours, allowed to cool to room temperature, then poured into water (200 ml) and partially concentrated in vacuo. The resulting precipitate was isolated by filtration, washed with water and dried in vacuo to give the title compound (23.56 g, 105%) as a white solid, which was used in the next step without purification.

1H NMR (400 MHz, DMSO) 1.44 (9H, s), 1.68-1.8 (2H, m), 2.19-2.36 (2H, m), 2.87 (2H, s ), 4.07 (2H, d),

- 52 043669

4.38 (1H, tt), 8.14 (1H, s), 11.63 (1H, s);

mass/charge MH+ 354.

Intermediate 88:

yl)piperidine-1-carboxylate.

tert-butyl-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purine-9-

M aq. NaOH solution (159 ml, 317.97 mmol) was added in one portion to tert-butyl 4-(2-chloro-8oxo-7,8-dihydro-9H-purin-9-yl)piperidin-1-carboxylate (22, 50 g, 63.59 mmol) and iodomethane (19.88 ml, 317.97 mmol) in THF (310 ml) at room temperature in air. The reaction mixture was stirred at room temperature for 3 hours, then partially concentrated in vacuo. The resulting precipitate was collected by filtration, washed with water and dried in vacuo at 45°C to obtain the title compound (17.98 g, 77%) as a white solid.

1H NMR (400 MHz, DMSO) 1.44 (9H, s), 1.7-1.78 (2H, m), 2.26 (2H, qd), 2.88 (2H, s), 3, 36 (3H, s), 4.07 (2H, d), 4.42 (1H, tt), 8.36 (1H, s);

mass/charge MH+ 368.

Intermediate 89: 2-chloro-7-methyl-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one hydrochloride.

M HCl in 1,4-dioxane (6.80 mL, 27.19 mmol) was added to tert-butyl-4-(2-chloro-7-methyl-8-oxo7,8-dihydro-9H-purine-9- yl)piperudin-1-carboxylate (2.0 g, 5.44 mmol) in methanol (25 ml) at room temperature and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. The resulting solid was triturated with EtOAc and a small amount of methanol to give the title compound (1.61 g, 97%) as the HCl salt.

1H NMR (400 MHz, DMSO) 1.97 (2H, d), 2.58 (2H, dd), 3.04-3.16 (2H, m), 3.37 (3H, s), 3. 41 (2H, d), 4.58 (1H, ddd), 8.39 (1H, s), 8.52 (1H, s), 9.08 (1H, s);

mass/charge MH+ 268.

Intermediate 90: 2-chloro-7-methyl-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin8-one.

Formaldehyde (37% in water) (0.20 ml, 2.72 mmol) was added in one portion to tert-butyl-4-(2-chloro7-methyl-8-oxo-7,8-dihydro-9H-purine-9 -yl)piperidin-1-carboxylate (500 mg, 1.36 mmol) in formic acid (2 ml) at room temperature. The reaction mixture was heated at 55°C for 18 hours, then allowed to cool to room temperature. concentrated in vacuo and absorbed sat. aq. a solution of NaHCO ₃ (20 ml) and EtOAc (20 ml). The organic layer was isolated and passed through a phase separation filter and concentrated in vacuo to give the title compound (200 mg, 52%) as a cream solid.

1H NMR (400 MHz, DMSO) 1.62-1.72 (2H, m), 1.94-2.04 (2H, m), 2.21 (3H, s), 2.45 (2H, td ), 2.85-2.93 (2H, m), 3.36 (3H, s), 4.1-4.23 (1H, m), 8.35 (1H, s);

mass/charge MH+ 282.

- 53 043669

Intermediate 91: 9-(1-acetylpiperidin-4-yl)-2-chloro-7-methyl-7,9-dihydro-8H-purin8-one.

Acetyl chloride (0.472 ml, 6.62 mmol) in DCM (5 ml) was added to triethylamine (2.51 ml, 18.04 mmol) and 2-chloro-7-methyl-9-(piperidin-4-yl)-7 ,9-dihydro-8H-purin-8-one hydrochloride (1.61 g, 5.29 mmol) in DCM (50 ml) at 0°C and the reaction mixture was stirred at room temperature for 1 hour. Reaction mixture diluted with DCM (50 ml), washed successively with water (50 ml) and sat. saline solution (40 ml). The organic layer was isolated and passed through a phase separation filter and concentrated in vacuo to give the title compound (1.36 g, 83%) as a beige solid.

^1H NMR (400 MHz, DMSO) 1.80 (2H, t), 2.06 (3H, s), 2.18 (1H, qd), 2.26-2.38 (1H, m), 2 .66 (1H, t), 3.20 (1H, t), 3.36 (3H, s), 3.96 (1H, d), 4.4-4.6 (2H, m), 8, 37 (1H, s);

mass/charge MN ⁺ 310.

Intermediate 92: tert-butyl (4-(benzylamino)-1-methylcyclohexyl)carbamate.

H ^Ph Μ ° ν

I n

Benzylamine (2.309 ml, 21.12 mmol) was added in one portion to tert-butyl (1-methyl-4oxocyclohexyl)carbamate (4 g, 17.60 mmol) in DCM (45 ml) at room temperature. The reaction mixture was stirred at RT for 2 hours. Sodium triacetoxyborohydride (7.46 g, 35.20 mmol) and AcOH (0.050 ml, 0.88 mmol) were added and the reaction mixture was stirred at RT for 16 hours, then quenched using saturation aq. Na ₂ CO ₃ solution (100 ml) and extracted with EtOAc (3x100 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in petroleum ether to give the title compound (5.60 g, 100%) as a colorless oil.

^1H NMR (300 MHz, CDCl3) 1.17-1.54 (15H, m), 1.73-1.89 (3H, m), 2.00-2.16 (1H, m), 2, 44-2.69 (1H, m), 3.80 (2H, d), 4.40-4.49 (1H, m), 7.18-7.41 (5H, m), NH protons not observed;

mass/charge MN ⁺ 319.

Intermediate 93: tert-butyl (4-amino-1-methylcyclohexyl)carbamate.

Pd/C 10% (1.00 g, 9.40 mmol) and tert-butyl (4-(benzylamino)-1-methylcyclohexyl)carbamate (5.60 g, 17.58 mmol) in ethanol (50 ml) stirred under hydrogen pressure of 3 atm. at RT for 30 hours. The reaction mixture was filtered through Celite® and concentrated in vacuo to give the title compound (4.06 g, 101%) as a pale yellow oil.

¹ H NMR (400 MHz, CDCl ₃ ) 1.26-1.34 (5H, m), 1.42 (9H, s), 1.59-1.86 (5H, m), 2.00-2 .12 (1H, m), 2.70-2.75 (1H, m), 3.45 (2H, s), 4.42 (1H, d).

Intermediate 94: Ethyl 4-((4-((tert-butoxycarbonyl)amino)-4methylcyclohexyl)amino)-2-chloropyrimidine-5-carboxylate.

ABOUT

Λ Λ

CI^N^NH

Φθ

Λ4 LΗ

DIPEA (3.69 mL, 21.12 mmol) was added dropwise to ethyl 2,4-dichloropyrimidine-5-carboxylate (3.89 g, 17.60 mmol) and tert-butyl-(4-amino-1- methylcyclohexyl)carbamate (4.02 g, 17.6 mmol) in acetonitrile (80 ml) at 0°C for a period of 2 minutes. The reaction mixture was stirred at room temperature for 16 hours and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 10% EtOAc in petroleum ether to give the title compound (6.0 g, 83%) as a pale yellow gum.

1H NMR (400 MHz, CDCl ₃ ) 1.33-1.43 (6H, m), 1.43-1.64 (11H, m), 1.70-1.82 (1H, m), 1, 85-2.01 (4H, m),

2.17 (1H, s), 4.07-4.24 (2H, m), 4.30-4.42 (2H, m), 8.24-8.57 (1H, m), 8, 67 (1H, s);

mass/charge MN ⁺ 413.

Intermediate 95: 4-((4-((tert-butoxycarbonyl)amino)-4-methylcyclohexyl)amino)2-chloropyrimidine-5-carboxylic acid.

LiOH (0.696 g, 29.06 mmol) was added in one portion to ethyl 4-((4-((tert-butoxycarbonyl)amino)4-methylcyclohexyl)amino)-2-chloropyrimidine-5-carboxylate (6.0 g, 14.5 mmol) in THF (50 ml) and water (50 ml) at 0°C. The reaction mixture was stirred at RT for 5 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting precipitate was isolated by filtration, washed with water (20 mL) and dried in vacuo to give the title compound (5.24 g, 94%) as a white solid.

1H NMR (400 MHz, DMSO) 1.16-1.63 (16H, m), 1.67-1.89 (3H, m), 2.08-2.18 (1H, m), 3.82 -4.08 (1H, m), 6.44 (1H, d), 8.56 (1H, s), 8.57-8.82 (1H, m);

mass/charge MN ⁺ 385.

Intermediate 96: tert-butyl (4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)-1methylcyclohexyl)carbamate.

Diphenylphosphoryl azide (2.91 mL, 13.51 mmol) was added in one portion to 4-((4-((tertbutoxycarbonyl)amino)-4-methylcyclohexyl)amino)-2-chloropyrimidine-5-carboxylic acid (5.20 g, 13.51 mmol) and triethylamine (1.88 ml, 13.51 mmol) in THF (50 ml) at room temperature. The reaction mixture was heated at 80°C for 2 days, then allowed to cool to room temperature and poured into water (150 ml). The resulting precipitate was isolated by filtration, washed with water (25 mL) and dried in vacuo to give the title compound (4.53 g, 88%) as a white solid.

1H NMR (400 MHz, DMSO) 1.24 (3H, d), 1.34-1.51 (10H, m), 1.58-1.80 (3H, m), 1.93 (1H, d ), 2.27-2.46 (3H, m), 4.07-4.20 (1H, m), 6.52 (1H, d), 8.12 (1H, d), 11.62 ( 1H, d);

mass/charge MN ⁺ 382.

Intermediate 97: tert-butyl-(4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)1-methylcyclohexyl).

DMF-DMA (2.01 mL, 15.0 mmol) was added in one portion to tert-butyl-(4-(2-chloro-8-oxo-7,8dihydro-9H-purin-9-yl)-1-methylcyclohexyl )carbamate (1.909 g, 5.00 mmol) in DMF (20 ml) at room temperature. The reaction mixture was heated at 80°C for 6 hours, allowed to cool to room temperature, then quenched with water (100 ml) and extracted with EtOAc (3x50 ml). The combined organic layers were dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 30% EtOAc in petroleum ether to give the title compound (1.540 g, 78%) as a pale yellow solid.

- 55 043669 ^1H NMR (400 MHz, CDCl ₃ ) 1.36-1.55 (14H, m), 1.59-1.80 (2H, m), 1.99-2.09 (1H, m ), 2.25-2.34 (1H, m),

2.49-2.66 (2H, m), 3.45 (3H, d), 4.29-4.47 (1H, m), 4.58 (1H, d), 8.01 (1H, d);

mass/charge MN ⁺ 396.

Intermediate 98: Ethyl 4-(((3S,4R)-1-(tert-butoxycarbonyl)-3-fluoropiperidin-4yl)amino)-2-chloropyrimidine-5-carboxylate.

DIPEA (5.19 mL, 29.78 mmol) was added portionwise to ethyl 2,4-dichloropyrimidine-5-carboxylate (5.06 g, 22.91 mmol) and tert-butyl-(3S,4R)-4- amino-3-fluoropiperidine-1-carboxylate (5.00 g, 22.91 mmol) in acetonitrile (100 ml) at 0°C. The reaction mixture was stirred at RT for 4 hours, then concentrated in vacuo, diluted with EtOAc (200 ml) and washed successively with water (100 ml) and sat. saline solution (100 ml). The organic layer was filtered through phase separation filter paper and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in n-heptane to give the title compound (4.87 g, 53%) as a white crystalline solid.

1H NMR (400 MHz, DMSO) 1.32 (3H, t), 1.41 (9H, s), 1.56-1.67 (1H, m), 1.82 (1H, d), 2, 93 (1H, s), 4.01 (1H, s), 4.33 (5H, q), 4.86 (1H, d), 8.53 (1H, d), 8.69 (1H, s );

mass/charge MN ⁺ 403.

Intermediate 99: 4-(((3S,4R)-1-(tert-butoxycarbonyl)-3-fluoropiperidin-4yl)amino)-2-chloropyrimidine-5-carboxylic acid.

Lithium hydroxide hydrate (0.99 g, 23.58 mmol) in water (45 ml) was added to ethyl 4-(((3S,4R)-1-(tert-butoxycarbonyl)-3-fluoropiperidin-4-yl)amino) -2-chloropyrimidine-5-carboxylate (4.75 g, 11.79 mmol) in THF (45 ml) at RT. The reaction mixture was stirred at RT for 1 h, then partially concentrated in vacuo and acidified using 2 M aq. HCl solution. The resulting precipitate was filtered, washed with water and dried to give the title compound (4.24 g, 96%) as a white solid.

1H NMR (400 MHz, DMSO) 1.41 (9H, s), 1.60 (2H, d), 1.82 (1H, d), 4.01 (1H, s), 4.18-4, 49 (3H, m), 4.85 (1H, d), 8.64 (1H, s), 8.79 (1H, s);

mass/charge MN ⁺ 375.

Intermediate 100: tert-butyl-(3S,4R)-4-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)3-fluoropiperidin-1-carboxylate.

Diphenylphosphoryl azide (2.44 ml, 11.31 mmol) was added to 4-(((3S,4R)-1-(tert-butoxycarbonyl)-3fluoropiperidin-4-yl)amino)-2-chloropyrimidine-5-carboxylic acid ( 4.24 g, 11.31 mmol) and triethylamine (1.58 ml, 11.31 mmol) in THF (75 ml) at room temperature. The reaction mixture was stirred at 80°C for 16 hours. The reaction mixture was allowed to cool to rt and water (150 ml) was added and partially concentrated in vacuo. The resulting solid was isolated by filtration and dried in vacuo to give the title compound (3.94 g, 94%) as a white solid.

- 56 043669

1H NMR (400 MHz, DMSO) 1.43 (9H, s), 1.82 (1H, d), 4.09-4.37 (4H, m), 4.53 (1H, ddd), 4, 82 (2Н, d),

8.17(1H, s), 11.71(1H, s);

mass/charge MN ⁺ 372.

Intermediate 101: tert-butyl-(3S,4R)-4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9Hpurin-9-yl)-3-fluoropiperidin-1-carboxylate.

M aq. NaOH solution (23.13 mL, 46.26 mmol) was added in one portion to tert-butyl-(3S,4R)-4(2-chloro-8-oxo-7,8-dihydro-9H-nurin-9-yl )-3-fluorineridine-1-carboxylate (3.44 g, 9.25 mmol) and iodomethane (2.89 ml, 46.26 mmol) in THF (50 ml) at room temperature in air. The reaction mixture was stirred at room temperature for 3 hours, then concentrated in vacuo and diluted with water (50 ml). The resulting precipitate was isolated by filtration and washed with water to give the title compound (2.08 g, 58%) as a pale orange solid.

1H NMR (400 MHz, DMSO) 1.43 (9H, s), 1.82 (1H, d), 3.03-3.15 (2H, m), 3.38 (3H, s), 4, 09-4.36 (3H, m), 4.58 (1H, ddt), 4.82 (1H, d), 8.39 (1H, s);

mass/charge MN ⁺ 386.

Intermediate 102: Ethyl 4-[[(3R)-1-tert-butoxycarbonylpyrrolidin-3-yl]amino]-2chloropyrimidine-5-carboxylate.

DIPEA (6.59 mL, 37.83 mmol) was added dropwise to a mixture of tert-butyl (R)-3-aminopyrrolidine-1carboxylate (5.42 g, 29.10 mmol) and ethyl 2,4-dichloropyrimidine 5-carboxylate (6.43 g, 29.10 mmol) in acetonitrile (108 ml) at -5°C for a period of 15 minutes under air. The reaction mixture was stirred for 2 hours, allowed to warm slowly to room temperature. The reaction mixture was concentrated in vacuo, diluted with EtOAc (100 ml) and washed with water, then sat. saline solution. The organic layer was dried over MgSO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc eluting with 0-70% EtOAc in n-heptane to give the title compound (7.92 g, 73%) as a pale yellow solid.

1H NMR (400 MHz, DMSO) 1.32 (3H, t), 1.41 (9H, s), 1.92-2.03 (1H, m), 2.19 (1H, s), 3, 21 (1H, dd), 3.37 (2H, t), 3.62 (1H, dd), 4.32 (2H, q), 4.59 (1H, s), 8.39 (1H, d ), 8.65 (1H, s);

mass/charge MN ⁺ 371.

Intermediate 103: 4-[[(3R)-1-tert-butoxycarbonylπpyrrolidin-3-yl]amino]-2chloropyrimidine-5-carboxylic acid.

Lithium hydroxide hydrate (1.79 g, 42.71 mmol) was added in one portion to ethyl 4-[[(3R)-1-t-butoxycarbonylpyrrolidin-3-yl]amino]-2-chloropyrimidine-5-carboxylate (7.92 g, 21.36 mmol) in THF (80 ml) and water (26.7 ml) at RT. The reaction mixture was stirred at RT for 3 hours, then partially concentrated in vacuo and acidified with 2 M aq. HCl solution. The resulting white solid was isolated by filtration and dried in vacuo at 45°C to give the title compound (7.07 g, 97%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.41 (9H, s), 1.95 (1H, s), 2.19 (1H, s), 3.20 (1H, dd), 3.37 (2H , t), 3.62 (1H,

- 57 043669 dd), 4.57 (1H, s), 8.61 (1H, s), 8.67 (1H, d), 13.80 (1H, s);

mass/charge MN ⁺ 343.

Intermediate 104: tert-butyl-(3R)-3-(2-chloro-8-oxo-7H-purin-9-yl)pyrrolidin-1-carboxylate.

Diphenylphosphoryl azide (4.44 mL, 20.63 mmol) was added in one portion to a solution of 4-[[(3R)-1-t-butoxycarbonylpyrrolidin-3-yl]amino]-2-chloropyrimidine-5-carboxylic acid (7.07 g, 20.63 mmol) and triethylamine (2.87 ml, 20.63 mmol) in THF (114 ml) at room temperature. The resulting solution was stirred at 80°C for 24 hours. The mixture was allowed to cool, then poured into water ( 100 ml), and no precipitate was formed. The solvent was removed in vacuo, resulting in the formation of a white precipitate in the water.

The precipitate was vacuum filtered, washed with water, air dried in vacuo for 30 minutes, then placed in a vacuum oven for 4 hours at 45°C to obtain the title compound (5.45 g, 78%) as a white solid substances that were used in the next step without purification.

1H NMR (400 MHz, DMSO) 1.42 (9H, d), 2.19 (1H, s), 2.50 (1H, s) 3.37 (1H, s), 3.53 - 3.67 (2H, m), 3.68-3.74 (1H, m), 4.94 (1H, q), 8.14 (1H, s), 11.64 (1H, s);

mass/charge [M-N] ^- 338.

Intermediate 105: tert-butyl (3R)-3-(2-chloro-7-methyl-8-oxopurin-9-yl)pyrrolidin 1-carboxylate.

M sodium hydroxide solution (36.8 ml, 73.6 mmol) was added in one portion to tert-butyl-(3R)-3(2-chloro-8-oxo-7H-purun-9-ul)nurrolidin-1-carboxylate (5.00 g, 14.72 mmol) and iodomethane (4.60 ml, 73.58 mmol) in THF (71.8 ml) at room temperature in air. The reaction mixture was stirred at RT for 3 hours, then partially concentrated in vacuo and extracted with DCM (100 ml). The organic layer was isolated and passed through a phase separation filter, then concentrated in vacuo to give the title compound (5.17 g, 99%) as a yellow gum.

^1H NMR (400 MHz, DMSO) 1.42 (9H, d), 2.21 (1H, s), 2.53 (1H, d), 3.36 (4H, s), 3.5-3 .75 (3H, m), 4.91-5.07 (1H, m), 8.37 (1H, s);

mass/charge MN ⁺ 354.

Intermediate 106: Ethyl 4-[[(38)-1-tert-butoxycarbonylpyrrolidin-3-yl]amino]-2chloropyrimidine-5-carboxylate.

Tert-butyl-(S)-3-aminopyrrolidine-1-carboxylate (5.00 g, 26.84 mmol) was slowly added to ethyl 2,4-dichloropyrimidine-5-carboxylate (5.93 g, 26.84 mmol ) and DIPEA (6.08 ml, 34.90 mmol) in acetonitrile (100 ml) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred at room temperature for 4 hours, then concentrated in vacuo, diluted with EtOAc (200 ml) and washed successively with water (100 ml) and sat. saline solution (100 ml). The organic layer was filtered through phase separation filter paper and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 50% EtOAc in n-heptane to give the title compound (5.40 g, 54%) as a white solid.

1H NMR (400 MHz, DMSO) 1.32 (3H, t), 1.41 (9H, s), 1.99 (1H, d), 2.19 (1H, s), 3.21 (1H, dd), 3.37 (2H, t), 3.62 (1H, dd), 4.32 (2H, q), 4.59 (1H, s), 8.39 (1H, d), 8, 65 (1H, s);

- 58 043669 mass/charge [M-N] ^- 369.

Intermediate 107: Chloropyrimidine-5-carboxylic acid.

4-[[(3S)-1 -tert-butoxycarbonylpyrrolidin-3-yl] amino] -2 -

Lithium hydroxide hydrate (1.22 g, 29.12 mmol) in water (50 ml) was added to ethyl 4-[[(38)-1-t-butoxycarbonylpyrrolidin-3-yl]amino]-2-chloropyrimidine-5-carboxylate (5.40 g, 14.56 mmol) in THF (50 ml) at RT. The reaction mixture was stirred at RT for 2 hours, then partially concentrated in vacuo, acidified with 2 M aq. HCl solution and extracted into DCM (100 ml). The organic layer was washed with brine (50 mL), passed through a phase separation filter paper and concentrated in vacuo to give the title compound (4.95 g, 99%).

1H NMR (400 MHz, DM8O) 1.41 (9H, s), 1.95 (1H, s), 2.19 (1H, s), 3.37 (2H, t), 3.59-3, 64 (2H, m), 4.57 (1H, s), 8.61 (2H, s), 13.80 (1H, s);

mass/charge MN ⁺ 343.

Intermediate 108: tert-butyl (38)-3-(2-chloro-8-oxo-7H-purin-9-yl)pyrrolidin-1-carboxylate.

Diphenylphosphoryl azide (3.08 ml, 14.29 mmol) was added to 4-[[(38)-1-t-butoxycarbonylpyrrolidin-3-yl]amino]-2-chloropyrimidine-5-carboxylic acid (4.90 g, 14.29 mmol) and triethylamine (1.99 ml, 14.29 mmol) in THF (50 ml) at room temperature. The reaction mixture was refluxed for 16 hours, allowed to cool to room temperature and diluted with water ( 100 ml). The resulting precipitate was isolated by filtration, washed with water and dried in vacuo to give the title compound (4.35 g, 90%) as a white solid.

1H NMR (400 MHz, DM8O) 1.41 (9H, s), 2.19 (1H, s), 3.37 (1H, d), 3.5-3.67 (3H, m), 3, 67-3.74 (1H, m), 4.89-5.01 (1H, m), 8.14 (1H, s), 11.66 (1H, s);

mass/charge MN ⁺ 340.

Intermediate 109: tert-butyl-(38)-3-(2-chloro-7-methyl-8-oxopurin-9-yl)pyrrolidin 1 -carboxylate.

Sodium hydride (60%) (0.662 g, 16.55 mmol) was added in portions to tert-butyl-(38)-3-(2-chloro-8oxo-7H-purin-9-yl)pyrrolidin-1-carboxylate (3 .75 g, 11.04 mmol) in DMF (30 ml) at room temperature. The reaction mixture was stirred for 1 hour, then cooled to 0°C and iodomethane (2.07 ml, 33.11 mmol) was added dropwise. . The reaction mixture was allowed to warm to room temperature, then stirred at room temperature for 1 hour. The reaction mixture was diluted with EtOAc (100 ml) and washed successively with water (3x50 ml) and sat. saline solution (50 ml). The organic layer was passed through filter paper to separate the phases and concentrated in vacuo. The resulting crude product was triturated with EtOAc to give the title compound (1.65 g, 42%) as a yellow solid.

1H NMR (400 MHz, DMSO) 1.42 (9H, d), 2.21 (1H, s), 2.47 (1H, d), 3.36 (4H, s), 3.61 (2H, d), 3.70 (1H, dd), 4.92-5.04 (1H, m), 8.37 (1H, s);

mass/charge MN ⁺ 354.

- 59 043669

Intermediate 110: tert-butyl-4-(7-methyl-2-((7-methylquinolin-6-yl)amino)-8-oxo7,8-dihydro-9H-purin-9-yl)piperidin-1- carboxylate.

Cesium carbonate (33.4 g, 102.4 mmol) was added to 7-methylquinoline-6-amine (5.40 g, 34.1 mmol) and tert-butyl-4-(2-chloro-7-methyl-8 -oxo-7,8-dihydro-9H-purin-9-yl)piperidin-1-carboxylate (12.56 g, 34.13 mmol) in 2-methyltetrahydrofuran (110 ml). The reaction mixture was bubbled with nitrogen for 5 min. 2,2'-bis(diphenylphosphanyl)-1,1'-binaphthalene (1.06 g, 1.71 mmol) and diacetoxypalladium (0.192 g, 0.85 mmol) were added and the reaction mixture was heated at 80°C for 1 h, then allowed to cool to room temperature. The reaction mixture was cooled to room temperature, filtered and the solid was washed with 10% MeOH in DCM (100 ml). The filtrate was concentrated in vacuo. The crude material was purified by fcc, 0 to 100% gradient elution (10% MeOH in EtOAc) in n-heptane to give the title compound (14.70 g, 88%) as a cream solid.

1H NMR (400 MHz, DMSO) 1.35 (9H, s), 1.78 (2H, d), 2.36 (2H, qd), 2.50 (3H, s), 2.84 (2H, s), 3.30 (3H, s), 4.05-4.17 (2H, m), 4.37 (1H, tt), 7.38 (1H, dd), 7.84 (1H, s ), 8.07-8.15 (1H, m), 8.16 (1H, s), 8.30 (1H, s), 8.51 (1H, s), 8.72 (1H, dd) ;

mass/charge MH+ 490.

Intermediate 111: tert-butyl-(3R)-3-[7-methyl-2-[(7-methyl-6-quinolyl)amino]-8oxopurin-9-yl]pyrrolidine-1-carboxylate.

Cesium carbonate (368 mg, 1.13 mmol) was added in one portion to 7-methylquinoline-6-amine (89 mg, 0.57 mmol) and tert-butyl-(3R)-3-(2-chloro-7-methyl -8-oxopurin-9-yl)pyrrolidin-1-carboxylate (200 mg, 0.57 mmol) in 1,4-dioxane (3.2 ml) at 20°C and degassed by bubbling nitrogen through the mixture for 5 min . Brettphos G3 precatalyst (25.6 mg, 0.03 mmol) was added and the reaction mixture was heated at 100° C. for 2 hours. The reaction mixture was filtered hot and the filter cup was washed with DCM (20 ml). The DCM layer was evaporated and the residue was absorbed onto silica gel, then purified by fcc (12 g Interchim column), eluting gradient from 0 to 5% MeOH in DCM, to give the title compound (174 mg, 64.7%) as a yellow gum .

^1H NMR (400 MHz, DMSO) 1.37 (9H, s), 2.18 (1H, d), 2.5 (3H, s), 2.64-2.76 (1H, m), 3 .33 (4H, s), 3.53 (1H, s), 3.64 (1H, s), 3.75 (1H, s), 4.96 (1H, s), 7.38 (1H, dd), 7.85 (1H, s), 8.12 (1H, s), 8.18 (1H, s), 8.30 (1H, d), 8.54 (1H, d), 8, 73 (1H, dd);

mass/charge MH+ 476.

Intermediate 112: tert-butyl-(3S)-3-[7-methyl-2-[(7-methyl-6-quinolyl)amino]-8oxopurin-9-yl]pyrrolidin-1-carboxylate.

Cesium carbonate (368 mg, 1.13 mmol) was added to 7-methylquinoline-6-amine (71.5 mg, 0.45 mmol) and tert-butyl-(3S)-3-(2-chloro-7-methyl -8-oxopurin-9-yl)pyrrolidin-1-carboxylate (200 mg, 0.57 mmol) in 1,4-dioxane (5 ml). Brettphos G3 precatalyst (25.6 mg, 0.03 mmol) was added and the reaction mixture was heated at 100°C for 1 hour. An additional 5% Brettphos G3 precatalyst was added and the reaction mixture was stirred for 1 hour, then allowed to cool to room temperature. t., filtered and the solid

- 60 043669 washed with DCM (10 ml). The combined organic layers were concentrated in vacuo and the resulting crude product was purified by fcc, elution gradient from 0 to 10% MeOH in

DCM to give the title compound (80 mg, 30%) as a cream solid.

^1H NMR (400 MHz, CDCl ₃ ) 1.47 (9H, d), 2.23 (1H, s), 2.59 (3H, s), 2.91 (1H, dq), 3.43 ( 4H, s), 3.66-3.91 (2H, m), 4.02 (1H, d), 5.10 (1H, s), 7.08 (1H, d), 7.31 (1H , s), 7.92 (1H, s), 7.97 (1H, s), 8.04 (1H, s), 8.69 (1H, d), 8.75 (1H, dd);

mass/charge MN ⁺ 476.

Intermediate 113: tert-butyl-4-(7-methyl-2-((7-methylcinnolin-6-yl)amino)-8-oxo7,8-dihydro-9H-purin-9-yl)piperidin-1- carboxylate.

Cesium carbonate (531 mg, 1.63 mmol) was added to 7-methylcinnolin-6-amine (130 mg, 0.82 mmol) and tert-butyl-4-(2-chloro-7-methyl-8-oxo-7 ,8-dihydro-9H-purin-9-yl)piperidin-1-carboxylate (300 mg, 0.82 mmol) in 1,4-dioxane (6 ml). Brettphos G3 precatalyst (37 mg, 0.04 mmol) was added and the reaction mixture was stirred at 100°C for 1 hour. An additional 5% precatalyst was added

Brettphos G3 and the reaction mixture was heated at 100°C for 2 hours. An additional 5% catalyst was added and the reaction mixture was heated at 100°C for 16 hours. An additional 5% catalyst was added and the reaction mixture was heated at 100°C for 1 hour. The reaction mixture was allowed to cool to room temperature, filtered and the solid washed with 10% MeOH in DCM (3 ml). The combined organic layers were concentrated in vacuo and the resulting crude mixture was purified by fcc, eluting with a gradient of 0 to 10% MeOH in DCM to give the title compound (138 mg, 35%) as a brown oil.

^1H NMR (400 MHz, DMSO) 1.33 (9H, s), 1.80 (2H, d), 2.08 (2H, s), 2.63 (3H, s), 2.88 (2H , s), 3.37 (3H, s), 4.11 (2H, s), 4.40 (1H, d), 7.88 (1H, d), 8.24 (1H, s), 8 .28 (1H, s), 8.52 (1H, s), 8.68 (1H, s), 9.10 (1H, d);

mass/charge MN ⁺ 491.

Intermediate 114: tert-butyl-(3S,4R)-3-fluoro-4-(7-methyl-2-((7-methylquinolin-6yl)amino)-8-oxo-7,8-dihydro-9H- purin-9-yl)piperidin-1-carboxylate.

Cesium carbonate (760 mg, 2.33 mmol) was added to 7-methylquinoline-6-amine (123 mg, 0.78 mmol) and tert-butyl-(3S,4R)-4-(2-chloro-7-methyl -8-oxo-7,8-dihydro-9H-purin-9-yl)-3-fluoropiperidine-1carboxylate (300 mg, 0.78 mmol) in 2-methyltetrahydrofuran (4 ml). The solution was degassed by bubbling nitrogen through the reaction mixture for 5 minutes. 2,2'-Bis(diphenylphosphanyl)1,1'-binaphthalene (24.2 mg, 0.04 mmol) and diacetoxypalladium (4.36 mg, 0.02 mmol) were added and the reaction mixture was heated at 80°C for 3 hours, then allowed to cool to room temperature, concentrated in vacuo, taken up with DCM (3 ml) and filtered. The filtrate was purified by fcc, eluting with a gradient of 0 to 10% methanol in DCM to give the title compound (132 mg, 33%) as a golden solid.

^1H NMR (400 MHz, DMSO) 1.21-1.51 (9H, m), 1.88 (1H, d), 2.96 (1H, s), 3.11-3.29 (2H, m), 3.30 (3H, s), 3.36 (3H, s), 4.17 (1H, s), 4.31 (1H, s), 4.53 (1H, dd), 4, 88 (1H, d), 7.36 (1H, dd), 7.83 (1H, s), 8.10 (1H, d), 8.21 (1H, s), 8.38 (1H, s ), 8.49 (1H, s), 8.72 (1H, dd);

mass/charge MN ⁺ 508.

- 61 043669

Intermediate 115: 7-methyl-2-[(7-methyl-6-quinolyl)amino]-9-[(3R)-pyrrolidin-3yl]purin-8-one.

M HCl in 1,4-dioxane (0.45 ml, 1.83 mmol) was added to tert-butyl-(3R)-3-[7-methyl-2-[(7-methyl-6quinolyl)amino]-8 -oxopurin-9-yl]pyrrolidin-1-carboxylate (174 mg, 0.37 mmol) in methanol (2.2 ml) at room temperature and the reaction mixture was stirred at room temperature for 1 hour. The mixture was diluted water (3 ml) to dissolve solids, then loaded onto a 5 g SCX column, washing with MeOH, then eluting with 1 N. NH ₃ in MeOH. The solvent was removed in vacuo to obtain the title compound (122 mg, 89%) as a yellow gum, which was directly used in the next step.

m/z [MH] ^- 374.

Intermediate 116: 7-methyl-2-[(7-methyl-6-quinolyl)amino]-9-[(3S)-pyrrolidin-3yl]purin-8-one.

M HCl in 1,4-dioxane (0.21 ml, 0.84 mmol) was added to tert-butyl-(3S)-3-[7-methyl-2-[(7-methyl-6quinolyl)amino]-8 -oxopurin-9-yl]pyrrolidin-1-carboxylate (80 mg, 0.17 mmol) in methanol (1 ml) at room temperature and the reaction mixture was stirred at room temperature for 2 hours, then concentrated in vacuo to obtain the HCl salt of the title compound (70 mg, 101%) as a pale yellow solid.

^1H NMR (400 MHz, DMSO) 2.32-2.41 (2H, m), 2.73 (3H, s), 3.32 (1H, dd), 3.37 (3H, s), 3 .57 (2H, q), 3.77 (1H, d), 5.11-5.22 (1H, m), 7.95 (1H, dd), 8.23 (1H, s), 8. 26 (1H, s), 8.78 (1H, s), 9.07 (2H, dd), 9.51 (2H, s), 9.84 (1H, s);

mass/charge [M-N] ^- 374.

Intermediate 117: 7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(piperidin-4-yl)7,9-dihydro-8H-purin-8-one.

M HCl in 1,4-dioxane (0.262 ml, 1.05 mmol) was added to tert-butyl-4-(7-methyl-2-((7methylcinnolin-6-yl)amino)-8-oxo-7,8 -dihydro-9H-purin-9-yl)piperidin-1-carboxylate (103 mg, 0.21 mmol) in methanol (1.2 ml) at room temperature and the reaction mixture was stirred at room temperature for 1 h, then concentrated in vacuo and loaded onto a 5 g SCX column, washing with MeOH, then eluting with 1 N. NH ₃ /MeOH. The solvent was removed in vacuo, MeCN (3 mL) was added and concentrated in vacuo to give the title compound (61 mg, 74%) as a brown solid.

1H NMR (400 MHz, DMSO) 1.75 (2H, d), 2.53 (2H, s), 2.63 (1H, s), 2.65 (3H, s), 3.1-3, 19 (3H, m), 3.37 (3H, s), 4.32 (1H, t), 8.08 (1H, d), 8.24 (1H, s), 8.28 (1H, s ), 8.63 (1H, s), 8.75 (1H, s), 9.12 (1H, d);

mass/charge MN ⁺ 391.

- 62 043669

Intermediate 118: Ethyl 4-(((3R,4R)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidin-3yl)amino)-2-chloropyrimidine-5-carboxylate.

DIPEA (7.33 mL, 41.97 mmol) was added in one portion to a stirred solution of ethyl 2,4dichloropyrimidine-5-carboxylate (4.64 g, 20.98 mmol) and tert-butyl-(3R,4R)-3 -amino-4-fluoropyrrolidine 1-carboxylate (obtained from a commercial source) (4.5 g, 22.03 mmol) in MeCN (100 ml) at room temperature. The reaction mixture was stirred at room temperature for 13 hours. the mixture was combined with a smaller amount of the reaction mixture using the same reagents and conditions (max. 2.26 mmol of desired product) and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 30% EtOAc in petroleum ether to give the title compound (7.60 g, 84% overall) as a white solid.

1H NMR (400 MHz, DMSO) 1.31 (3H, q), 1.43 (9H, s), 3.39-3.80 (4H, m), 4.21-4.38 (2H, m ), 4.70 (1H, s), 5.26 (1H, d), 8.34 (1H, d), 8.69 (1H, s);

mass/charge MN ⁺ 389.

Intermediate 119: 4-(((3R,4R)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidin-3yl)amino)-2-chloropyrimidine-5-carboxylic acid.

A solution of lithium hydroxide (0.91 g, 38.06 mmol) in water (150 ml) was added in one portion to a stirred solution of ethyl 4-(((3R,4R)-1-(tert-butoxycarbonyl)-4-fluoropyrrolidine- 3-yl)amino)-2chloropyrimidine-5-carboxylate (7.40 g, 19.03 mmol) in THF (150 ml) at room temperature. The reaction mixture was stirred at room temperature for 16 hours, then concentrated in vacuo, diluted with water (100 ml) and acidified to pH 2 using 2 M aq. HCl solution. The resulting precipitate was collected and dried under vacuum to give the title compound (6.10 g, 89%) as a white solid.

1H NMR (400 MHz, DMSO) 1.43 (9H, s), 3.39-3.81 (5H, m), 4.67 (1H, s), 5.25 (1H, d), 8. 64 (1H, s), 13.78 (1H, br s);

mass/charge MN ⁺ 361.

Intermediate 120: tert-butyl-(3R,4R)-3-(2-chloro-8-oxo-7,8-dihydro-9H-purin-9-yl)4-fluoropyrrolidin-1-carboxylate.

Diphenylphosphonic acid azide (3.57 g, 12.97 mmol) was added in one portion to 4-(((3R,4R)-1(tert-butoxycarbonyl)-4-fluoropyrrolidin-3-yl)amino)-2-chloropyrimidine- 5-carboxylic acid (3.90 g, 10.81 mmol) and Et ₃ N (4.52 ml, 32.43 mmol) in toluene (140 ml) at room temperature. The reaction mixture was heated at 100°C for 16 h, then allowed to cool to room temperature. The reaction mixture was combined with a smaller amount of the reaction mixture using the same reagents and conditions (max. 4.16 mmol of desired product) and the resulting mixture was diluted with DCM (200 ml) . The organic layer was isolated and washed with sat. saline solution, then dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 5% MeOH in DCM to give the title compound (3.50 g, 67% overall) as a brown oil.

1H NMR (400 MHz, DMSO) 1.43 (9H, s), 3.53-4.06 (4H, m), 5.03 (1H, d), 5.44-5.64 (1H, m ), 8.18 (1H, s), 11.82 (1H, s);

mass/charge MN ⁺ 358.

- 63 043669

Intermediate 121: tert-butyl-(3R,4R)-3-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9Hpurin-9-yl)-4-fluoropyrrolidin-1-carboxylate.

60% w/w sodium hydride in mineral oil (0.67 g, 16.77 mmol) was added portionwise to tert-butyl-(3R,4R)-3-(2-chloro-8-oxo-7,8-dihydro-9H -purin-9-yl)-4-fluoropyrrolidin-1-carboxylate (4.00 g, 11.18 mmol) in DMF (100 ml) at 0°C and the reaction mixture was stirred at 0°C for 10 minutes. Iodomethane (1.40 mL, 22.36 mmol) was added and the reaction mixture was stirred at RT for 1 hour. The reaction mixture was combined with a smaller amount of the reaction mixture using the same reagents and conditions (max. 1.40 mmol of the required product). The resulting mixture was poured into water (1 L) and the resulting precipitate was collected by filtration and dried under vacuum, then triturated with MTBE and purified by fcc, eluting gradient from 0 to 3% MeOH in DCM to give the title compound (1.80 g , 39% overall) as a yellow solid.

^1H NMR (400 MHz, DMSO) 1.43 (9H, s), 3.36 (3H, s), 3.51-4.02 (4H, m), 5.06 (1H, d), 5 .36-5.68 (1H, m), 8.40 (1H, s);

mass/charge MN ⁺ 372.

Intermediate 122: tert-butyl-(3R,4R)-3-fluoro-4-(7-methyl-2-((7-methylquinolin-6yl)amino)-8-oxo-7,8-dihydro-9H- purin-9-yl)pyrrolidin-1-carboxylate.

Brettphos Pd G3 (0.51 g, 0.56 mmol) was added in one portion to a stirred mixture of tert-butyl(3R,4R)-3-(2-chloro-7-methyl-8-oxo-7,8-dihydro- 9H-purin-9-yl)-4-fluoropyrrolidine-1-carboxylate (1.60 g, 4.30 mmol), CS ₂ CO ₃ (3.51 g, 10.76 mmol) and 7-methylquinoline-6- amine (0.72 g, 4.52 mmol) in 1,4-dioxane (60 ml) at room temperature. The reaction mixture was heated at 100°C for 16 hours. The mixture was allowed to cool to room temperature and diluted with using DCM (200 ml). The resulting mixture was washed with sat. saline solution, and the resulting organic layer was isolated and dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0 to 2% MeOH in DCM to give the title compound (1.80 g, 85%) as a yellow solid.

^1H NMR (400 MHz, DMSO) 1.35 (9H, s), 3.30 (3H, s), 3.31 (3H, s), 3.22-3.94 (4H, m), 5 .02 (1H, d), 5.57 (1H, ddt), 7.38 (1H, dd), 7.83 (1H, s), 8.06-8.30 (3H, m), 8. 62 (1H, d), 8.72 (1H, dd);

mass/charge MN ⁺ 494.

Example 1: 9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one.

Cesium carbonate (210 mg, 0.65 mmol) was added to 9-(1-acetylpiperidin-4-yl)-2-chloro-7-methyl-7,9dihydrO-8H-pURin-8-oneU (100 mg, 0. 32 mmol) and 7-methylcinnolin-6-amine (51.4 mg, 0.32 mmol) in 1,4-dioxane (3 ml). Brettphos Pd G3 (14.6 mg,

0.02 mmol) and the resulting suspension was stirred at 100°C for 1 hour. An additional 5% Brettphos Pd G3 catalyst was added and the reaction mixture was heated at 100°C for 2 hours, then an additional 5% Pd catalyst was added and the reaction mixture heated at 100°C for 16 hours. The reaction mixture was allowed to cool to room temperature and concentrated in vacuo. The residue was taken up in DMF (2 ml), filtered and purified by preparative HPLC. The residue was diluted in DCM (1 ml) and a few drops of n-heptane were added. The solvent was removed in vacuo to give yellow

- 64 043669 oil. Acetonitrile (2 mL) was added and the solid was isolated by filtration and dried in vacuo to give the title compound (8.0 mg, 6%) as a yellow solid.

1H NMR (400 MHz, DMSO) 1.84 (2H, s), 1.95 (3H, s), 2.62 (5H, s), 3.1-3.24 (2H, m), 3, 36 (3H, s), 3.96 (1H, d), 4.48 (1H, t), 4.59 (1H, d), 7.86 (1H, d), 8.23 (1H, s ), 8.27 (1H, s), 8.49 (1H, s), 8.73 (1H, s), 9.09 (1H,

d);

mass/charge MN ⁺ 433.

Example 2: 9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro8H-purin-8-one.

O ⁰

U-N

Α.. Α ,Ν n |

Cesium carbonate (210 mg, 0.65 mmol) was added to 9-(1-acetylpiperidin-4-yl)-2-chloro-7-methyl-7,9dihydro-8H-purin-8-one (100 mg, 0. 32 mmol) and 7-methylquinoxaline-6-amine (51.4 mg, 0.32 mmol) in 1,4-dioxane (3 ml). Brettphos Pd G3 (14.6 mg, 0.02 mmol) was added and the resulting suspension was stirred at 100°C for 1 hour. An additional 5 mol% Brettphos Pd G3 (14.6 mg, 0.02 mmol) was added and the reaction the mixture was heated at 100°C for 2 hours. An additional 5 mol% Brettphos Pd G3 (14.6 mg, 0.02 mmol) was added and the reaction mixture was heated at 100°C for 1 hour. The reaction mixture was allowed to cool to ... and concentrated in vacuo. The residue was diluted with DMF (2 ml), filtered and purified by preparative HPLC. DCM (2 ml) and a few drops of diethyl ether were added and the mixture was concentrated in vacuo to give the title compound (27 mg, 19%) as a green solid.

1H NMR (400 MHz, DMSO) 1.76 (1H, d), 1.85 (1H, d), 1.98 (3H, s), 2.15-2.27 (1H, m), 2, 55 (5H, s), 3.15 (1H, t), 3.34 (3H, s), 3.96 (1H, d), 4.39-4.49 (1H, m), 4.52 (1H, d), 7.89 (1H, s), 8.22 (1H, s), 8.55 (1H, s), 8.66 (1H, s), 8.73 (1H, d) , 8.76 (1H, d);

mass/charge MN ⁺ 433.

Example 3: 9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinazolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one.

RuPhos Pd G3 (9.2 mg, 11 µmol) was added to 9-(1-acetylpiperidin-4-yl)-2-chloro-7-methyl-7,9dihydro-8H-purin-8-one (68.1 mg , 0.22 mmol), 7-methylquinazoline-6-amine (35 mg, 0.22 mmol), RuPhos (10.3 mg, 0.02 mmol) and CS ₂ CO ₃ (215 mg, 0.66 mmol) in 1,4-dioxane (1 ml) under nitrogen atmosphere. The reaction mixture was heated at 100°C for 16 hours, then allowed to cool to room temperature and concentrated in vacuo and purified by preparative HPLC to give the title compound (22 mg, 23%) as a white solid.

1H NMR (400 MHz, DMSO) 1.74-1.87 (2H, m), 1.95 (3H, s), 2.16-2.31 (1H, m), 2.34-2.49 (1H, m), 2.53-2.67 (4H, m), 3.09-3.21 (1H, m), 3.34 (3H, s), 3.95 (1H, d), 4.38-4.48 (1H, m), 4.48-4.58 (1H, m), 7.86 (1H, s), 8.20 (1H, s), 8.43 (1H, s), 8.78 (1H, s), 9.13 (1H, s), 9.36 (1H, s);

mass/charge MN ⁺ 433.

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Example 4: 9-(1-acetylpiperidin-4-yl)-2-((2,7-dimethylquinoxalin-6-yl)amino)-7-methyl-7,9dihydro-8H-purin-8-one.

Example 5: 9-(1-acetylpiperidin-4-yl)-2-((3,7-dimethylquinoxalin-6-yl)amino)-7-methyl-7,9dihydro-8H-purin-8-one.

Cesium carbonate (1.05 g, 3.23 mmol) was added to 9-(1-acetylpiperidin-4-yl)-2-chloro-7-methyl-7,9dihydro-8H-purin-8-one (500 mg, 1.61 mmol), a mixture of 2,7-dimethylquinoxalin-6-amine (224 mg, 1.29 mmol) and 3,7-dimethylquinoxalin-6-amine (55.9 mg, 0.32 mmol) in 1.4 -dioxane (10 ml). Brettphos Pd G3 (73.2 mg, 0.08 mmol) was added and the reaction mixture was heated at 100°C for 1 hour. An additional 5 mol% Brettphos Pd G3 (73.2 mg, 0.08 mmol) was added and the reaction the mixture was heated at 100°C for 1 hour, then allowed to cool to room temperature, filtered and washed with DCM (10 ml). The filtrate was concentrated in vacuo and purified by fcc, eluting with a gradient of 0 to 10% MeOH in DCM to give the title compounds as a mixture. Examples 4 and 5 were separated by SFC to obtain the title compounds 4 (62 mg, 10%) as a cream solid and 5 (20 mg, 3%) as a cream solid.

Example 4: ^1H NMR (400 MHz, DMSO) 1.75 (1H, d), 1.84 (1H, d), 1.98 (3H, s), 2.21 (1H, qd), 2. 45 (1H, dd), 2.52 (3H, s), 2.54-2.64 (1H, m), 2.66 (3H, s), 3.13 (1H, d), 3.34 (3H, s), 3.96 (1H, d), 4.44 (1H, tt), 4.52 (1H, d), 7.77-7.81 (1H, m), 8.19 ( 1H, s), 8.45 (1H, s), 8.61 (1H, s), 8.68 (1H, s);

mass/charge MH+447.

Example 5: 1H NMR (400 MHz, DMSO) 1.77 (1H, d), 1.85 (1H, d), 1.94 (3H, s), 2.15-2.27 (1H, m) , 2.4-2.48 (1H, m), 2.52 (3H, s), 2.57-2.64 (1H, m), 2.66 (3H, s), 3.13 (1H , d), 3.34 (3H, s), 3.94 (1H, d), 4.45 (1H, ddd), 4.53 (1H, d), 7.83 (1H, s), 8 .20 (1H, s), 8.39 (1H, s), 8.63 (2H, d);

mass/charge MH+447.

Example 6: 9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro8H-purin-8-one.

RuPhos Pd G3 (40.1 mg, 0.04 mmol) was added to 2-chloro-9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl7,9-dihydro-8H-purin-8-one ( 125 mg, 0.44 mmol), 7-methylquinoline-6-amine (70 mg, 0.44 mmol) and CS2CO3 (288 mg, 0.88 mmol) in 1,4-dioxane (4 ml). The reaction mixture was heated at 100°C for 16 hours, then allowed to cool to room temperature, filtered and the solid washed with MeOH (10 ml). The combined organic layers were concentrated in vacuo and purified by preparative HPLC to give the title compound (20 mg, 11%) as a white solid.

1H NMR (300 MHz, DMSO) 1.18-1.37 (2H, m), 1.67-1.78 (2H, m), 1.87-1.98 (2H, m), 2.28 -2.43 (2H, m), 2.51 (3H, s), 3.32 (3H, s), 3.35-3.37 (1H, m), 4.06-4.24 (1H , m), 4.66 (1H, d), 7.42 (1H, dd), 7.85 (1H, s), 8.08-8.14 (1H, m), 8.14 - 8, 21 (1H, m), 8.29 (1H, s), 8.61 (1H, s), 8.74 (1H, dd);

mass/charge MH+ 405.

Example 7: 9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)αmino)-7,9dihydro-8H-purin-8-one.

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Cesium carbonate (230 mg, 0.71 mmol) was added to 2-chloro-9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl7,9-dihydro-8H-purin-8-one (100 mg, 0.35 mmol) and 7-methylcinnolin-6-amine (56.3 mg, 0.35 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (32 mg, 0.04 mmol) was added. The reaction mixture was heated at 100°C for 2 hours. Additional 5% Brettphos Pd G3 was added and stirred for 1 hour. The reaction mixture was allowed to cool to room temperature and concentrated. The resulting crude product was purified by preparative HPLC. The pure fractions were combined and partially concentrated and the product crystallized from water where it was isolated by filtration and dried in vacuo to give the title compound (30 mg, 21%) as a beige solid.

1H NMR (400 MHz, DMSO) 1.32 (2H, q), 1.76 (2H, d), 1.96 (2H, d), 2.3-2.44 (2H, m), 2, 64 (3H, s), 3.35 (3H, s), 3.43 (1H, s), 4.20 (1H, ddd), 4.67 (1H, d), 7.88 (1H, d ), 8.26 (2H, s), 8.52 (1H, s), 8.71 (1H, s), 9.15 (1H, d);

mass/charge MN ⁺ 406.

Example 8: 2-((4,7-dimethylquinolin-6-yl)amino)-9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl-7,9dihydro-8H-purun-8-one.

Cesium carbonate (488 mg, 1.50 mmol) was added in one portion to 4,7-dimethylquinoline-6-amine (129 mg, 0.75 mmol) and 2-chloro-9-((1r,4r)-4-hydroxycyclohexyl )-7-methyl-7,9-dihydro-8H-purin-8-one (212 mg, 0.75 mmol) in 1,4-dioxane (5 ml) at room temperature and the reaction mixture was degassed by bubbling nitrogen through the mixture for 5 minutes. Brettphos Pd G3 (67.9 mg, 0.07 mmol) was added and the reaction mixture was heated at 100°C for 4 hours, then allowed to cool to room temperature, diluted with DCM and filtered through celite. The DCM layer was concentrated in vacuo and purified by fcc, eluting gradient from 0 to 5% MeOH in DCM to give the title compound (175 mg, 56%) as a cream solid.

1H NMR (400 MHz, DMSO) 1.26 (2H, q), 1.71 (2H, d), 1.90 (2H, d), 2.22-2.38 (2H, m), 2, 50 (3H, s), 2.59-2.66 (3H, m), 3.32 (3H, s), 3.33-3.37 (1H, m), 4.18 (1H, ddd) , 4.61 (1H, d), 7.26-7.31 (1H, m), 7.84 (1H, s), 8.14 (1H, s), 8.40 (1H, s), 8.56(1H, s), 8.60(1H, d);

mass/charge MN ⁺ 419.

Example 9: 9-(( 1 r,4r)-4-hydroxycyclohexyl)-2-((4-methoxy-7-methylquinolin-6-yl)amino)-7-methyl7,9-dihydro-8H-purine-8 -He.

Cesium carbonate (346 mg, 1.06 mmol) was added in one portion to 4-methoxy-7-methylquinoline-6amine (100 mg, 0.53 mmol) and 2-chloro-9-((1r,4r)-4-hydroxycyclohexyl )-7-methyl-7,9-dihydro-8H-purin-8one (150 mg, 0.53 mmol) in 1,4-dioxane (4 ml) at RT and the reaction mixture was degassed by bubbling nitrogen through the mixture within 5 min. Brettphos Pd G3 (48.1 mg, 0.05 mmol) was added and the reaction mixture was heated at 100°C for 18 hours, then allowed to cool to room temperature, diluted with DCM and filtered through celite. The DCM layer was concentrated in vacuo and purified by fcc, eluting gradient from 0 to 5% MeOH in DCM to give the title compound (70 mg, 30%) as a cream solid.

^1H NMR (400 MHz, DMSO) 1.26 (2H, q), 1.70 (2H, d), 1.89 (2H, d), 2.32 (3H, q), 2.48 (3H , s), 3.32 (3H, s), 4.02 (3H, s), 4.09-4.19 (1H, m), 4.58 (1H, d), 6.92 (1H, d), 7.78 (1H, s), 8.14 (1H, s), 8.46 (1H, s), 8.51 (1H, s), 8.58 (1H, d);

mass/charge MN ⁺ 435.

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Example 10: 9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9dihydro-8H-purin-8-one.

Cesium carbonate (1.73 g, 5.31 mmol) was added to 2-chloro-9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl7,9-dihydro-8H-purin-8-one (750 mg, 2.65 mmol) and 7-methylquinoxaline-6-amine (422 mg, 2.65 mmol) in 1,4-dioxane (15 ml). Brettphos Pd G3 (120 mg, 0.13 mmol) was added and the reaction mixture was heated at 100°C for 2 hours, then allowed to cool to room temperature, filtered and the solid washed with DCM (10 ml). The combined DCM layers were concentrated in vacuo and purified by fcc, eluting gradient from 0 to 10% MeOH in DCM. The resulting solid was suspended in MeCN (50 ml). The suspension was refluxed, then allowed to cool to room temperature, filtered, washed with a little MeCN and the solid dried at 45°C in vacuo to give the title compound (460 mg, 43%) as a yellow solid substances.

^1H NMR (400 MHz, DMSO) 1.29 (2H, q), 1.73 (2H, d), 1.94 (2H, d), 2.41 (2H, d), 2.58 (3H , s), 3.34 (3H, s), 3.54 (1H, d), 4.13-4.24 (1H, m), 4.60 (1H, d), 7.90 (1H, s), 8.22 (1H, s), 8.61 (1H, s), 8.69 (1H, s), 8.73 (1H, d), 8.80 (1H, d);

mass/charge MN ⁺ 406.

Example 11: 9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8H-purin-8-one.

BrettPhos Pd G3 (64.1 mg, 0.07 mmol) was added to 2-chloro-9-((1s,4s)-4-hydroxycyclohexyl)-7methyl-7,9-dihydro-8H-purin-8-one ( 100 mg, 0.35 mmol), 7-methylquinoline-6-amine (84 mg, 0.53 mmol) and CS ₂ CO ₃ (346 mg, 1.06 mmol) in 1,4-dioxane (3 ml). The reaction mixture was heated at 100°C for 16 hours. The reaction mixture was cooled to room temperature and directly purified by fcc, elution gradient from 0 to 35% MeCN in water (with 0.1% FA), then further purified by preparative HPLC to obtain the title compound (45 mg, 32%) as a white solid.

¹ H NMR (400 MHz, DMSO) 1.47-1.61 (4H, m), 1.79-1.89 (2H, m), 2.52 (3H, s), 2.67-2, 82 (2H, m), 3.33 (3H, s), 3.87-3.94 (1H, m), 4.15-4.28 (1H, m), 4.48 (1H, d) , 7.38 (1H, dd), 7.83 (1H, s), 8.16 (1H, s), 8.32 (1H, dd), 8.38 (1H, s), 8.47 ( 1H, s), 8.70 (1H, dd);

mass/charge MN ⁺ 405.

Example 12: 9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-7,9dihydro-8H-purin-8-one.

Cesium carbonate (230 mg, 0.71 mmol) was added to 2-chloro-9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl7,9-dihydro-8H-purin-8-one (100 mg, 0.35 mmol) and 7-methylcinnolin-6-amine (56.3 mg, 0.35 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (16.0 mg, 0.02 mmol) was added. The reaction mixture was heated at 100°C for 2 hours. An additional 5% Pd catalyst was added and the reaction mixture was heated at 100°C for 2 hours, then allowed to cool to room temperature, concentrated in vacuo and purified by preparative HPLC. then further purified by trituration with MeCN to give the title compound (35 mg, 24%) as a beige solid.

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1H NMR (400 MHz, DMSO) 1.58 (4H, q), 1.87 (2H, d), 2.65 (3H, s), 2.79 (2H, q), 3.37 (3H, s), 3.95 (1H,

s), 4.25 (1H, t), 4.56 (1H, d), 8.18 (1H, d), 8.23 (1H, s), 8.28 (1H, s), 8, 49 (1H, s), 8.74 (1H, s), 9.10 (1H, d);

mass/charge MH+ 406.

Example 13: 9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9dihydro-8H-purin-8-one.

HE

Cesium carbonate (230 mg, 0.71 mmol) was added to 2-chloro-9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl7,9-dihydro-8H-purin-8-one (100 mg, 0.35 mmol) and 7-methylquinoxaline-6-amine (56.3 mg, 0.35 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (16.0 mg, 0.02 mmol) was added. The reaction mixture was heated at 100°C for 2 hours. Additional Brettphos Pd G3 (16.0 mg, 0.02 mmol) was added and the reaction mixture was then heated at 100°C for 1 hour, then allowed to cool to room temperature. and concentrated in vacuo. The residue was diluted with MeOH (3 ml), filtered and purified by preparative HPLC. Pure fractions were combined and partially concentrated. The resulting precipitate was isolated by filtration and dried in vacuo to give the title compound (65 mg, 45%) as a yellow solid.

1H NMR (400 MHz, DMSO) 1.45-1.61 (4H, m), 1.83 (2H, d), 2.58 (3H, s), 2.66-2.79 (2H, m ), 3.34 (3H, s), 3.91 (1H, s), 4.25 (1H, td), 4.34 (1H, d), 7.90 (1H, s), 8.20 (1H, s), 8.54 (1H, s), 8.70 (1H, s), 8.73 (1H, d), 8.79 (1H, d);

mass/charge MN ⁺ 406.

Example 14: 9-((1s,4s)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)αmino)7,9-dihydro-8H-purine-8 -He.

Example 15: 9-((1 r,4r)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)7,9-dihydro-8H-purine- 8-on.

RuPhos Pd G3 (169 mg, 0.20 mmol) and 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (94 mg, 0.20 mmol) were added to 2-chloro-9-(4 -hydroxy-4-methylcyclohexyl)-7-methyl-7,9-dihydro-8Hpurin-8-one (300 mg, 1.01 mmol), 7-methylquinolin-6-amine (240 mg, 1.52 mmol) and CS ₂ CO ₃ (988 mg, 3.03 mmol) in 1,4-dioxane (5 ml). The reaction mixture was heated at 100°C for 5 hours, then allowed to cool to room temperature and directly purified by fcc, elution gradient from 0 to 32% MeCN in water (with 0.1% FA), then further purified by preparative HPLC to give 14 (102 mg, 24%) as a white solid and 15 (36 mg, 9%) as a white solid.

Example 14: 1H NMR (400 MHz, DMSO) 1.17 (3H, s), 1.37-1.56 (4H, m), 1.66-1.75 (2H, m), 2.52 ( 3H, s), 2.65-2.80 (2H, m), 3.32 (3H, s), 4.11-4.24 (2H, m), 7.39 (1H, dd), 7 .83 (1H, s), 8.15 (1H, s), 8.31 (1H, dd), 8.38 (1H, s), 8.45 (1H, s), 8.71 (1H, dd);

mass/charge MH+419.

Example 15: 1H NMR (400 MHz, DMSO) 0.69 (3H, s), 1.35-1.48 (2H, m), 1.48-1.62 (4H, m), 2.18- 2.34 (2H, m), 2.46 (3H, s), 3.31 (3H, s), 4.03-4.16 (1H, m), 4.31 (1H, s), 7 .41 (1H, dd), 7.85 (1H, s), 8.08 (1H, s), 8.15 (2H, d), 8.68 (1H, s), 8.74 (1H, dd);

mass/charge MN ⁺ 419.

- 69 043669

Example 16: 9-((1 s,4s)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)7,9-dihydro-8H-purine- 8-on.

Example 17: 9-((1 g,4 g)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)7,9-dihydro-8H-purine- 8-on.

Cesium carbonate (220 mg, 0.67 mmol) was added to 2-chloro-9-((1s,4s)-4-hydroxy-4methylcyclohexyl)-7-methyl-7,9-dihydro-8H-purin-8-one (100 mg, 0.34 mmol) and 7-methylcinnoline-6amine (53.6 mg, 0.34 mmol) in 1,4-dioxane (2 ml). Brettphos Pd G3 (15.3 mg, 0.02 mmol) was added and the reaction mixture was heated at 100°C for 2 hours. An additional 5% catalyst was added and the reaction mixture was heated at 100°C for 2 hours, then allowed to cool to KT and concentrated in vacuo. The residue was diluted with DCM (3 ml), filtered and purified by fcc, eluting with a gradient of 0 to 10% MeOH in DCM to give Example 16 (15 mg, 11%) as a light brown solid and Example 17 (25 mg, 18 %) as a beige solid.

Example 16: ^1H NMR (400 MHz, DMSO) 1.19 (3H, s), 1.50 (4H, q), 1.73 (2H, d), 2.64 (3H, s), 2. 76 (2H, q), 3.36 (3H, s), 4.21 (2H, s), 8.15 (1H, d), 8.22 (1H, s), 8.27 (1H, s ), 8.47 (1H, s), 8.72 (1H, s), 9.10 (1H, d);

mass/charge MN ⁺ 420.

Example 17: 1H NMR (400 MHz, DMSO) 0.85 (3H, s), 1.42-1.54 (2H, m), 1.57-1.69 (4H, m), 2.27- 2.4 (2H, m), 2.57-2.64 (3H, m), 3.35 (3H, s), 4.17 (1H, dq), 4.34 (1H, s), 7 .92 (1H, dd), 8.25 (2H, d), 8.33 (1H, s), 8.79 (1H, s), 9.15 (1H, d);

mass/charge MN ⁺ 420.

Example 18: 9-((1g,4g)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6yl)amino)-7,9-dihydro-8H-nurine-8- He.

Example 19: 9-((1s,4s)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6yl)amino)-7,9-dihydro-8H-purine-8- He.

Cesium carbonate (220 mg, 0.67 mmol) was added to 2-chloro-9-((1r,4r)-4-hydroxy-4-methylcyclohexyl)7-methyl-7,9-dihydro-8H-πpurine-8- on (100 mg, 0.34 mmol) and 7-methylquinoxaline-6-amine (53.6 mg, 0.34 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed and Brettphos Pd G3 (15.3 mg, 0.02 mmol) was added. The reaction mixture was heated at 100°C for 2 hours. An additional 5 mol% Brettphos Pd G3 (15.3 mg, 0.02 mmol) was added and the reaction mixture was heated at 100°C for 2 hours, then allowed to cool to ... and concentrated in vacuo. The residue was diluted with MeOH (3 ml), filtered and purified by preparative HPLC. The pure fractions were combined and partially concentrated in vacuo, and the resulting precipitate was isolated by filtration and dried in vacuo to give Example 18 (35 mg, 25%) as a yellow solid and Example 19 (55 mg, 39%) as a beige solid. .

Example 18: 1H NMR (400 MHz, DMSO) 0.82 (3H, s), 1.4-1.51 (2H, m), 1.59 (4H, t), 2.22-2.4 ( 2H, m), 2.54 (3H, s), 3.34 (3H, s), 4.14 (1H, ddt), 4.30 (1H, s), 7.88-7.95 (1H , m), 8.21 (1H, s), 8.41 (1H, s), 8.71 (1H, s), 8.75 (1H, d), 8.79 (1H, d);

mass/charge MN ⁺ 420.

Example 19: 1H NMR (400 MHz, DMSO) 1.16 (3H, s), 1.38-1.55 (4H, m), 1.68 (2H, d), 2.57 (3H, s) , 2.66-2.78 (2H, m), 3.33 (3H, s), 4.10 (1H, s), 4.20 (1H, td), 7.89 (1H, s), 8.19 (1H, s), 8.50 (1H, s), 8.69-8.74 (2H, m), 8.78 (1H, d);

mass/charge MN ⁺ 420.

- 70 043669

Example 20: 9-((1s,4s)-4-hydroxy-1-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6yl)amino)-7,9-dihydro-8H-purine-8- He.

RuPhos Pd G3 (14.1 mg, 0.02 mmol) was added to 2-chloro-9-((1s,4s)-4-hydroxy-1-methylcyclohexyl)7-methyl-7,9-dihydro-8H-purine -8-one (50 mg, 0.17 mmol), 7-methylquinoxaline-6-amine (26.8 mg, 0.17 mmol), CS ₂ CO ₃ (453 mg, 1.39 mmol) and RuPhos (15 .7 mg, 0.03 mmol) in 1,4-dioxane (2 ml). The reaction mixture was heated at 100°C for 16 hours and then concentrated in vacuo. The residue was purified by fcc, eluting with a gradient of 0 to 50% MeCN in water to give the title compound (45 mg, 64%) as a yellow solid.

^1H NMR (400 MHz, DMSO) 1.44-1.56 (5H, m), 1.56-1.66 (2H, m), 1.99-2.11 (2H, m), 2, 57 (3H, s), 2.90-2.95 (2H, m), 3.30 (3H, s), 3.62-3.68 (1H, m), 4.47 (1H, s) , 7.89 (1H, s), 8.22 (1H, s), 8.51 (1H, s), 8.56 (1H, s), 8.73 (1H, d), 8.79 ( 1H, d);

mass/charge MN ⁺ 420.

Example 21: (8)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro8H-purin-8-one.

ABOUT

ABOUT \__/

V-N'' r^N

VJ fj

H I

RuPhos Pd G3 (263 mg, 0.31 mmol) and 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (147 mg, 0.31 mmol) were added to 2-chloro-7-methyl- 9-[(3S)-tetrahydro-3-furanyl]-7,9-dihydro-8H-purin8-one (960 mg, 3.77 mmol), 7-methylcinnolin-6-amine (500 mg, 3.14 mmol ) and CS2CO3 (3.07 g, 9.42 mmol) in 1,4-dioxane (15 ml). The reaction mixture was heated at 100°C for 16 h, then allowed to cool to room temperature and directly purified by fcc, eluting gradient from 0 to 40% MeCN in water, then further purified by preparative HPLC to give the title compound ( 847 mg, 72%) as a light yellow solid.

1H NMR (300 MHz, DMSO) 2.27-2.35 (1H, m), 2.36-2.48 (1H, m), 2.62 (3H, s), 3.35 (3H, s ), 3.74-3.88 (1H, m), 3.92-4.19 (3H, m), 4.97-5.15 (1H, m), 7.94 (1H, d), 8.22 (1H, s), 8.27 (1H, s), 8.63 (1H, s), 8.67 (1H, s), 9.13 (1H, d);

mass/charge MN ⁺ 378.

Form A.

The final product, (8)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9dihydro-8H-purin-8-one, was analyzed by XRPD and D8C and found it to be crystalline. Using XRPD of a sample of the material, the diffraction pattern shown in FIG. 3. Form A (8)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8Hpurin-8-one is characterized by with at least one peak at a 29 value of 9.7° or 12.9° as measured using CuKa radiation. The ten most pronounced XRPD peaks are shown in Table. A.

- 71 043669

Table A

Ten most prominent XRPD peaks for Form A (8)-7-methyl-2-((7methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purine-8 -she

Angle 2-theta (2Θ) Intensity, % 9.7 100.0 12.9 14.7 17.9 P,9 12.5 By 21.0 By 17.6 10.9 19.4 10.4 26.4 6.5 26.0 5.5 15.8 5.1

where 2-theta values are ±0.2°.

Example 22: (S)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro8H-purin-8-one.

RuPhos Pd G3 (32.8 mg, 0.04 mmol) and 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (18.3 mg, 0.04 mmol) were added to 2-chloro- 7-methyl-9-[(3S)-tetrahydro-3-furanyl]-7,9-dihydro-8H-purin8-one (100 mg, 0.39 mmol), 7-methylquinoxaline-6-amine (68.8 mg, 0.43 mmol) and CS ₂ CO ₃ (384 mg, 1.18 mmol) in 1,4-dioxane (2 ml). The reaction mixture was heated at 100°C for 16 hours. The reaction mixture was allowed to cool to room temperature and directly purified by fcc, eluting gradient from 0 to 70% MeCN in water, then further purified by preparative HPLC to give the title compound (44 mg, 30%) as a yellow solid.

1H NMR (300 MHz, DMSO) 2.18-2.32 (1H, m), 2.51-2.62 (4H, m), 3.35 (3H, s), 3.76-3.84 (1H, m), 3.88 (1H, dd), 3.97 (1H, t), 4.11 (1H, q), 4.92-5.05 (1H, m), 7.91 ( 1H, d), 8.24 (1H, s), 8.58 (1H, s), 8.66 (1H, s), 8.74 (1H, d), 8.80 (1H, d);

mass/charge MN ⁺ 378.

Example 23: (R)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro8H-purin-8-one.

RuPhos Pd G3 (26.3 mg, 0.03 mmol) and 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (14.7 mg, 0.03 mmol) were added to 2-chloro- 7-methyl-9-[(3R)-tetrahydro-3-furanyl]-7,9-dihydro-8H-purin8-one (96 mg, 0.38 mmol), 7-methylcinnolin-6-amine (50 mg, 0.31 mmol) and CS ₂ CO ₃ (205 mg, 0.63 mmol) in 1,4-dioxane (1.5 ml). The reaction mixture was heated at 100°C for 16 hours. The reaction mixture was allowed to cool to room temperature and directly purified by fcc, eluting gradient from 0 to 40% MeCN in water, then further purified by preparative HPLC to give the title compound (60 mg, 51%) as a pink solid.

^1H NMR (400 MHz, DMSO) 2.21-2.35 (1H, m), 2.38-2.51 (1H, m), 2.64 (3H, d), 3.37 (3H, s), 3.76-3.86 (1H, m), 3.99-4.15 (3H, m), 5.02-5.14 (1H, m), 7.96 (1H, d) , 8.24 (1H, s), 8.30 (1H, s), 8.69 (2H, d), 9.15 (1H, d);

mass/charge MN ⁺ 378.

- 72 043669

Example 24: (R)-7-methyl-2-((7-methylquinoxalin-6-ul)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro8H-purin-8-one.

RuPhos Pd G3 (32.8 mg, 0.04 mmol) and 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (18.32 mg, 0.04 mmol) were added to 2-chloro- 7-methyl-9-[(3R)-tetrahydro-3-furanyl]-7,9-dihydro-8Hpurin-8-one (100 mg, 0.39 mmol), 7-methylquinoxalin-6-amine (68.8 mg, 0.43 mmol) and CS ₂ CO ₃ (256 mg, 0.79 mmol) in 1,4-dioxane (3 ml). The reaction mixture was heated at 100°C for 3 hours. The reaction mixture was allowed to cool to room temperature and directly purified by fcc, gradient elution of 0-34% MeCN in water, then further purified by preparative HPLC to give the title compound ( 71 mg, 48%) as a white solid.

^1H NMR (400 MHz, DMSO) 2.14-2.33 (1H, m), 2.50-2.62 (4H, m), 3.33 (3H, s), 3.72-4, 02 (3H, m), 4.03-4.17 (1H, m), 4.89-5.05 (1H, m), 7.89 (1H, d), 8.23 (1H, s) , 8.57 (1H, s), 8.64 (1H, s), 8.72 (1H, d), 8.78 (1H, d);

mass/charge MN ⁺ 378.

Example 25: (R)-7-methyl-2-((7-methylcinnolin-6-ul)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9dihydro-8H-purine-8- He.

Cesium carbonate (243 mg, 0.74 mmol) was added to 2-chloro-7-methyl-9-[(3K)-tetrahydropyran-3yl]purin-8-one (100 mg, 0.37 mmol) and 7-methylcinnoline -6-amine (59.2 mg, 0.37 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (16.9 mg, 0.02 mmol) was added. The reaction mixture was heated at 100°C for 2 hours. An additional 5% Pd catalyst was added and the reaction mixture was stirred at 100°C for 2 hours. The reaction mixture was allowed to cool to room temperature and concentrated in vacuo. The residue was diluted with DMF (3 ml), filtered and purified by preparative HPLC to give the title compound (30 mg, 21%) as a brown solid.

1H NMR (400 MHz, DMSO) 1.70 (1H, d), 1.79 (1H, d), 1.97 (1H, d), 2.64 (3H, s), 3.2-3, 3 (2H, m), 3.36 (3H, s), 3.85 (2H, d), 3.99 (1H, t), 4.29-4.4 (1H, m), 7.92 (1H, d), 8.27 (2H, s), 8.52 (1H, s), 8.76 (1H, s), 9.16 (1H, d);

mass/charge MN ⁺ 392.

Example 26: (R)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9dihydro-8H-purine-8- He.

Cesium carbonate (243 mg, 0.74 mmol) was added to 2-chloro-7-methyl-9-[(3R)-tetrahydropuran-3yl]purin-8-one (100 mg, 0.37 mmol) and 7-methylquinoxaline -6-amine (59.2 mg, 0.37 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (16.9 mg, 0.02 mmol) was added. The reaction mixture was heated at 100°C for 2 hours. An additional 5 mol% Brettphos G3 precatalyst (16.87 mg, 0.02 mmol) was added and the reaction mixture was heated at 100°C for 2 hours. The reaction mixture was allowed to cool to KT and concentrated in vacuo. The residue was diluted with DMF (3 ml), filtered and purified by preparative HPLC. The pure fractions were combined and partially concentrated and the resulting precipitate was isolated by filtration and dried in vacuo to give the title compound (50 mg, 34%).

1H NMR (400 MHz, DMSO) 1.71 (2H, dt), 1.94 (1H, d), 2.59 (4H, s), 3.35 (3H, s), 3.41 (1H, td), 3.79-3.9

- 73 043669 (2H, m), 4.05 (1H, t), 4.33 (1H, ddt), 7.91 (1H, s), 8.25 (1H, s), 8.65 (1H , s), 8.7-8.76 (2H, m), 8.79 (1H, d);

mass/charge MN ⁺ 392.

Example 27: (S)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9dihydro-8H-purine-8- He.

⁰ VZ°

V-N r^N 'Δ Η I

Cesium carbonate (1.21 g, 3.72 mmol) was added to 2-chloro-7-methyl-9-[(3S)-tetrahydropyran-3yl]purin-8-one (500 mg, 1.86 mmol) and 7 -methylcinnolin-6-amine (296 mg, 1.86 mmol) in 1,4-dioxane (12 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (84 mg, 0.09 mmol) was added. The reaction mixture was heated at 100°C for 24 hours. The reaction mixture was filtered while hot, then the solid was washed with DCM (10 ml). The combined organic layers were concentrated in vacuo. The resulting crude product was purified by fcc, eluting gradient from 0 to 5% MeOH in DCM, then further purified by preparative HPLC. The resulting solid was stirred overnight in MeCN then filtered to give the title compound (153 mg, 21%) as a light brown solid.

^1H NMR (400 MHz, DMSO) 1.63-1.83 (2H, m), 1.97 (1H, d), 2.53-2.58 (1H, m), 2.64 (3H, s), 3.25 (1H, td), 3.35 (3H, s), 3.82-3.9 (2H, m), 3.99 (1H, t), 4.34 (1H, ddt ), 7.91 (1H, dd), 8.26 (2H, d), 8.52 (1H, s), 8.75 (1H, s), 9.15 (1H, d); ₊ MN mass/charge ⁺ 392.

Example 28: (S)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9dihydro-8H-purine-8- He.

° ίΖ? ⁰ Ύι Α ^Η I

Cesium carbonate (243 mg, 0.74 mmol) was added to 2-chloro-7-methyl-9-[(3S)-tetrahydropyran-3yl]purin-8-one (100 mg, 0.37 mmol) and 7-methylquinoxaline -6-amine (59.2 mg, 0.37 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (16.9 mg, 0.02 mmol) was added. The reaction mixture was heated at 100°C for 2 hours. An additional 5 mol% Brettphos G3 precatalyst (16.9 mg, 0.02 mmol) was added and the reaction mixture was heated at 100°C for 2 hours. The reaction mixture was allowed to cool to KT and concentrated in vacuo. The residue was diluted with DMF (3 ml), filtered and purified by preparative HPLC. The combined pure fractions were partially concentrated in vacuo and the resulting solid isolated by filtration and dried in vacuo to give the title compound (65 mg, 45%) as a light brown solid.

1H NMR (400 MHz, DMSO) 1.70 (2H, dt), 1.93 (1H, d), 2.59 (4H, s), 3.35 (3H, s), 3.36-3, 46 (1H, m), 3.8-3.9 (2H, m), 4.05 (1H, t), 4.33 (1H, ddd), 7.91 (1H, s), 8.25 (1H, s), 8.66 (1H, s), 8.71-8.75 (2H, m), 8.79 (1H, d);

mass/charge MN ⁺ 392.

Example 29: 7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro8H-purin-8-one.

Cesium carbonate (243 mg, 0.74 mmol) was added to 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9dihydro-8H-purin-8-one (100 mg, 0.37 mmol) and 7-methylcinnolin-6-amine (59.2 mg, 0.37 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (16.9 mg, 0.02 mmol) was added. The reaction mixture was heated at 100°C for 2 hours. An additional 5% catalyst was added and the reaction mixture was heated at 100°C for 2 hours, then allowed to cool to room temperature and

- 74 043669 concentrated in vacuo. The residue was diluted with MeOH (3 ml), filtered and purified by preparative HPLC then trituration with MeCN to give the title compound (15 mg, 10%) as a beige solid.

1H NMR (400 MHz, DMSO) 1.75 (2H, d), 2.58-2.71 (5H, m), 3.37 (3H, s), 3.49 (2H, t), 4, 03 (2H, dd), 4.51 (1H, ddd), 7.95 (1H, d), 8.25 (1H, s), 8.29 (1H, s), 8.70 (2H, d ), 9.14 (1H, d);

mass/charge MH+392.

Example 30: 7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(tetrahydro-2H-πpyran-4-yl)-7,9-dihydro8H-pURin-8-one.

RuPhos Pd G3 (218 mg, 0.24 mmol) was added to 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'biphenyl (112 mg, 0.24 mmol), 7-methylquinoline-6-amine (380 mg, 2.40 mmol), 2-chloro-7-methyl-9(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purun-8-one (645 mg, 2.40 mmol ) and CS ₂ CO ₃ (1.56 g, 4.80 mmol) in 1,4-dioxane (4 ml). The reaction mixture was heated at 100°C for 16 hours, then allowed to cool to room temperature, filtered and the solid washed with MeOH (10 ml). The combined organic layers were concentrated in vacuo and purified by preparative HPLC to give the title compound (208 mg, 22%) as a white solid.

1H NMR (300 MHz, DMSO) 1.64-1.76 (2H, m), 2.52 (3H, s), 2.54-2.71 (2H, m), 3.33 (3H, s ), 3.44 (2H, dd), 3.92-4.04 (2H, m), 4.36-4.53 (1H, m), 7.41 (1H, dd), 7.84 ( 1H, s), 8.17 (1H, s), 8.19 (1H, d), 8.44 (1H, s), 8.55 (1H, s), 8.71 (1H, dd);

mass/charge MN ⁺ 391.

Example 31: 7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9dihydro-8H-purin-8-one.

Cesium carbonate (1.81 g, 5.58 mmol) was added to 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9dihydro-8H-purin-8-one (750 mg, 2.79 mmol) and 7-methylquinoxaline-6-amine (444 mg, 2.79 mmol) in 1,4-dioxane (2 ml). Brettphos Pd G3 (127 mg, 0.14 mmol) was added and the reaction mixture was heated at 100°C for 2 hours, then allowed to cool to room temperature, filtered and the solid washed with DCM (10 ml). The combined organic layers were concentrated in vacuo and purified by fcc, eluting gradient from 0 to 10% MeOH in DCM, and triturated with MeCN to give the title compound (340 mg, 31%) as a pale yellow solid.

^1H NMR (400 MHz, DMSO) 1.68-1.75 (2H, m), 2.54-2.63 (5H, m), 3.35 (3H, s), 3.43 (2H, t), 3.98 (2H, dd), 4.46 (1H, ddt), 7.91 (1H, s), 8.22 (1H, s), 8.60 (1H, s), 8, 65 (1H, s), 8.74 (1H, d), 8.79 (1H, d);

mass/charge MH+392.

Example 32: 7-methyl-2-((7-methylquinazolin-6-yl)amino)-9-(tetrahydro-2H-niran-4-yl)-7,9-dihydro8H-purin-8-one.

RuPhos Pd G3 (9.2 mg, 11.0 µmol) was added to 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9dihydro-8H-purin-8-one ( 59.1 mg, 0.22 mmol), 7-methylquinazoline-6-amine (35 mg, 0.22 mmol), RuPhos (10.3 mg, 0.02 mmol) and CS ₂ CO ₃ (215 mg, 0 .66 mmol) in 1,4-dioxane (1 ml). The reaction mixture was heated at 100°C for 16 hours, then allowed to cool to room temperature and concentrated in vacuo. The resulting crude product was purified by fcc, eluting gradient from 0 to 40% MeCN in water as eluent with 0.1% NH4HCO3, then further purified by preparative HPLC to obtain the title compound (16 mg, 19%) as a white solid substances.

- 75 043669

1H NMR (400 MHz, DMSO) 1.68-1.77 (2H, m), 2.54-2.70 (5H, m), 3.35 (3H, s), 3.41-3.52 (2Н, m),

3.96-4.05 (2H, m), 4.41-4.54 (1H, m), 7.87 (1H, s), 8.23 (1H, s), 8.67 (1H, s), 8.73 (1H, s), 9.12 (1H, s), 9.40 (1H, s);

mass/charge MH+ 392.

Example 33: 2-((2,7-dimethylquinoxalin-6-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9dihydro-8H-purin-8-one.

Example 34: 2-((3,7-dimethylquinoxalin-6-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9dihydro-8H-purin-8-one.

Cesium carbonate (1.21 mg, 3.72 mmol) was added to 2-chloro-7-methyl-9-(tetrahydro-2H-pyran-4-yl)7,9-dihydro-8H-purin-8-one ( 500 mg, 1.86 mmol), a mixture of 2,7-dimethylquinoxalin-6-amine (258 mg, 1.49 mmol) and 3,7-dimethylquinoxalin-6-amine (64.5 mg, 0.37 mmol) in 1,4-dioxane (10 ml). Brettphos Pd G3 (84 mg, 0.09 mmol) was added and the reaction mixture was heated at 100°C for 1 hour. An additional 5 mol% Brettphos G3 precatalyst (84 mg, 0.09 mmol) was added and the reaction mixture was stirred for 1 hour then allowed to cool to room temperature, filtered and the solid was washed with DCM (10 ml). The combined filtrate was concentrated in vacuo and the crude product was purified by fcc, eluting with a gradient of 0 to 10% MeOH in DCM, then trituration with EtOAc to give a mixture of the title compounds. The isomers were separated by SFC to give 33 (125 mg, 17%) as a cream solid and 34 (25 mg, 3%) as a cream solid.

Example 33: 1H NMR (400 MHz, DMSO) 1.66-1.74 (2H, m), 2.53-2.62 (5H, m), 2.66 (3H, s), 3.34 ( 3H, s), 3.42 (2H, t), 3.97 (2H, dd), 4.45 (1H, tt), 7.76-7.82 (1H, m), 8.18 (1H , s), 8.48 (1H, s), 8.60 (1H, s), 8.70 (1H, s);

mass/charge MH+406.

Example 34: 1H NMR (400 MHz, DMSO) 1.71 (2H, d), 2.53-2.63 (5H, m), 2.65 (3H, s), 3.35 (3H, s) , 3.43 (2H, t), 3.98 (2H, dd), 4.46 (1H, ddd), 7.84 (1H, s), 8.20 (1H, s), 8.51 ( 1H, s), 8.60 (1H, s), 8.64 (1H, s);

mass/charge MH+ 406.

Example 35: 9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8H-purine-8- He.

RuPhos Pd G3 (26.4 mg, 0.03 mmol) was added to 2-chloro-9-(1,1-dioxidotetrahydro-2H-thiopyran-4yl)-7-methyl-7,9-dihydro-8H-purine- 8-one (100 mg, 0.32 mmol), 7-methylquinoline-6-amine (59.9 mg, 0.38 mmol), RuPhos (14.7 mg, 0.03 mmol) and CS2CO3 (309 mg, 0.95 mmol) in 1,4-dioxane (2 ml). The reaction mixture was heated at 100°C for 16 hours. The reaction was cooled to room temperature and the mixture was purified by flash chromatography with a C18 flash column, elution gradient from 0 to 50% MeCN in water, then further purified by preparative HPLC to obtain the title compound (42 mg, 30%) as a white solid.

^1H NMR (400 MHz, DMSO) 2.14 (2H, d), 2.52 (3H, s), 3.00 (2H, q), 3.17 (2H, d), 3.34 (3H , s), 3.42-3.55 (2H, m), 4.60-4.73 (1H, m), 7.39 (1H, dd), 7.85 (1H, s), 8, 20 (1H, s), 8.30-8.33 (1H, m), 8.34 (1H, s), 8.46 (1H, s), 8.72 (1H, dd);

mass/charge MH+ 439.

Example 36: 7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(oxetan-3-yl)-7,9-dihydro-8H-purin-8-one.

Cesium carbonate (271 mg, 0.83 mmol) was added to 2-chloro-7-methyl-9-(oxetan-3-yl)-7,9-dihydro-8H-76 043669 purin-8-one (100 mg, 0.42 mmol) and 7-methylquinoline-6-amine (65.7 mg, 0.42 mmol) in 1,4-dioxane (2 ml). The reaction mixture was degassed with nitrogen and Brettphos Pd G3 (18.8 mg, 0.02 mmol) was added. The reaction mixture was heated at 100°C for 1 hour. An additional 5% Pd catalyst was added and the reaction mixture was stirred for 1 hour at 100°C. An additional 2 equivalents of cesium carbonate and 5% Pd catalyst were added and the reaction mixture was heated at 100°C for 18 hours, then allowed to cool to room temperature and concentrated in vacuo. The residue was diluted with DCM (3 mL), filtered and purified by fcc, eluting with a gradient of 0 to 8% MeOH in DCM to give the title compound (15 mg, 10%) as an off-white solid.

^1H NMR (400 MHz, DMSO) 2.52 (3H, d), 3.33 (3H, s), 4.83 (2H, dd), 5.36 (2H, t), 5.49 (1H , qn), 7.39 (1H, dd), 7.84 (1H, s), 8.17 (1H, d), 8.21 (1H, s), 8.51 (1H, s), 8 .57 (1H, s), 8.71 (1H, dd);

mass/charge MH+ 363.

Example 37: 7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one.

M HCl in 1,4-dioxane (0.23 ml, 0.92 mmol) was added to tert-butyl-4-(7-methyl-2-((7methylquinolin-6-yl)amino)-8-oxo-7 ,8-dihydro-9H-purin-9-yl)piperidin-1-carboxylate (90 mg, 0.18 mmol) in methanol (1 ml) at room temperature and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed in vacuo and the residue was loaded onto a 5 g SCX column, washing with MeOH, then eluting with 1 N. NH ₃ /MeOH. The solvent was removed in vacuo and MeCN (2 mL) was added and concentrated in vacuo to give the title compound (62 mg, 87%) as a yellow solid.

1H NMR (400 MHz, DMSO) 1.71 (2H, d), 2.45 (2H, dt), 2.53 (3H, s), 2.61 (2H, t), 3.10 (2H, d), 3.33 (3H, s), 4.27 (1H, ddd), 7.40 (1H, dd), 7.85 (1H, s), 8.16 (1H, s), 8, 27 (1H, d), 8.47 (2H, d), 8.72 (1H, dd);

mass/charge MH+ 390.

Example 38: 9-((3S,4R)-3-fluoropiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8H-purine-8- He.

M HCl in 1,4-dioxane (1.00 mL, 4.00 mmol) was added to m-butyl-(3S,4R)-3-fluoro-4-(7-methyl-2((7-methylquinoline-6 -yl)amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)piperidin-1-carboxylate (85 mg, 0.17 mmol) in methanol (3 ml) at room temperature and the reaction mixture was stirred at room temperature for 1 hour, then concentrated in vacuo. 1M ammonia in methanol (1.5 mL) and water (1 mL) were added and the resulting solid was isolated by filtration to give the title compound (50 mg, 73%) as a white solid.

1H NMR (400 MHz, DMSO) 1.77 (1H, d), 2.06 (1H, s), 2.52 (3H, s), 2.62 (1H, d), 2.82 (1H, dd), 3.04 (1H, d), 3.20 (2H, d), 3.36 (3H, s), 4.35-4.5 (1H, m), 4.73 (1H, d ), 7.40 (1H, dd), 7.84 (1H, s), 8.20 (2H, s), 8.47 (2H, d), 8.72 (1H, dd);

mass/charge MH+ 408.

Example 39: 9-((1s,4s)-4-amino-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8H-purine-8- He.

Example 40: 9-((1r,4r)-4-amino-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8H-purine-8- He.

3rd generation precatalyst RuPhos (84 mg, 0.10 mmol) and RuPhos (47.1 mg, 0.10 mmol) adding

- 77 043669 li to tert-butyl-(4-(2-chloro-7-methyl-8-oxo-7,8-dihydro-9H-purin-9-yl)-1-methylcyclohexyl) (400 mg, 1, 01 mmol), 7-methylquinoline-6-amine (240 mg, 1.52 mmol) and CS2CO3 (988 mg, 3.03 mmol) in 1,4-dioxane (5 ml). The reaction mixture was stirred at 100°C for 16 hours, then allowed to cool to room temperature and concentrated in vacuo to give tert-butyl-(1-methyl-4-(7-methyl-2-((7-methylquinoline- 6yl)amino)-8-oxo-7,8-dihydro-9H-purin-9-yl)cyclohexyl)carbamate as a yellow resin, which was used without further purification. 4 M HCl in 1,4-dioxane (10 ml, 40 mmol) was added dropwise to the crude mixture and the reaction mixture was stirred at room temperature for 3 hours, then directly purified by flash chromatography with a C18 flash column. eluting gradient from 0 to 70% MeCN in water, then further purified by preparative HPLC to give Example 39 (48 mg, 11%) as a white solid and Example 40 (46 mg, 11%) as a yellow solid.

Example 39: ^1H NMR (300 MHz, DMSO) 1.03 (3H, s), 1.32-1.59 (6H, m), 2.49 (4H, s), 2.51-2.59 (1H, m), 3.32 (3H, s), 4.02-4.19 (1H, m), 7.40 (1H, dd), 7.84 (1H, s), 8.14 ( 1H, s), 8.19 (1H, d), 8.23 (1H, s), 8.60 (1H, s), 8.73 (1H, dd);

mass/charge MN ⁺ 418.

Example 40: 1H NMR (300 MHz, DMSO) 0.61 (3H, s), 1.20-1.39 (2H, m), 1.39-1.59 (4H, m), 2.18- 2.39 (2H, m), 2.46 (3H, s), 3.31 (3H, s), 3.98-4.16 (1H, m), 7.40 (1H, dd), 7 .84 (1H, s), 8.08 (1H, s), 8.15 (2H, d), 8.67 (1H, s), 8.74 (1H, dd);

mass/charge MN ⁺ 418.

Example 41: (R)-7-methyl-9-(1-methylpyrrolidin-3-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro8H-purin-8-one.

Sodium triacetoxyborohydride (138 mg, 0.65 mmol) was added to 7-methyl-2-[(7-methyl-6quinolyl)amino]-9-[(3R)-pyrrolidin-3-yl]purin-8-one (122 mg, 0.32 mmol), formaldehyde (37% aq. solution) (0.048 ml, 0.65 mmol) and acetic acid (0.074 ml, 1.30 mmol) in DCM (1 ml) at room temperature and reaction the mixture was stirred at room temperature for 24 hours, then concentrated in vacuo. The residue was purified by preparative HPLC to give the title compound (32 mg, 25%) as a white solid.

1H NMR (400 MHz, DMSO) 2.07-2.18 (4H, m), 2.27 (1H, ddd), 2.41 (1H, q), 2.48 (3H, s), 2, 59-2.7 (2H, m), 2.90 (1H, t), 3.32 (3H, s), 4.85 (1H, dtd), 7.41 (1H, dd), 7.86 (1H, s), 8.15 (1H, s), 8.17-8.22 (2H, m), 8.58 (1H, s), 8.74 (1H, dd);

mass/charge [M-N] ^- 388.

Example 42: (S)-7-methyl-9-(1-methylpyrrolidin-3-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro8H-purin-8-one.

Sodium triacetoxyborohydride (51.5 mg, 0.24 mmol) was added to a mixture of formaldehyde (37% solution in water, 19.7 mg, 0.24 mmol) and HCl salt of 7-methyl-2-[(7-methyl- 6-quinolol)amino]-9-[(3S)-pyrrolidin3-yl]purin-8-one (50 mg, 0.12 mmol) in MeOH at room temperature. The reaction mixture was stirred at room temperature for 30 min. An additional 2 eq. was added. sodium triacetoxyborohydride (51.5 mg, 0.24 mmol) and the reaction mixture was stirred for 30 min, concentrated in vacuo and purified by preparative HPLC to give the title compound (20 mg, 42%) as a white solid.

1H NMR (400 MHz, DMSO) 2.13 (4H, s), 2.22-2.31 (1H, m), 2.41 (1H, q), 2.48 - 2.49 (3H, m ), 2.64 (2H, dq), 2.90 (1H, t), 3.32 (3H, s), 4.85 (1H, dtd), 7.41 (1H, dd), 7.86 (1H, s), 8.15 (1H, s), 8.19 (2H, d), 8.58 (1H, s), 8.74 (1H, dd);

mass/charge MN ⁺ 390.

- 78 043669

Example 43: 7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(1-methylpiperudin-4-yl)-7,9-dihydro-8Hpurin-8-one.

Sodium triacetoxyborohydride (60.8 mg, 0.29 mmol) was added to 7-methyl-2-((7-methylcinnolin-6yl)amino)-9-(piperidin-4-yl)-7,9-dihydro-8H- purin-8-one (56 mg, 0.14 mmol), formaldehyde (37% aq. solution) (0.021 ml, 0.29 mmol) and acetic acid (0.033 ml, 0.57 mmol) in DCM (1 ml) at RT and the reaction mixture was stirred for 3.5 hours, then concentrated in vacuo and purified by preparative HPLC to obtain the title compound (22 mg, 38%) as a white solid.

1H NMR (400 MHz, DMSO) 1.73 (2H, d), 2-2.08 (3H, m), 2.29 (3H, s), 2.64-2.66 (3H, m), 2.72 (1H, d), 2.95 (2H, d), 3.37 (3H, s), 4.17-4.27 (1H, m), 8.07 (1H, d), 8 .24 (1H, s), 8.29 (1H, s), 8.64 (1H, s), 8.84 (1H, s), 9.14 (1H, d);

mass/charge MN ⁺ 405.

Example 44: 7-methyl-9-(1-methylpiperudin-4-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one.

Paraformaldehyde (568 mg, 1.46 mmol) was added to 7-methyl-2-((7-methylquinolin-6-yl)amino)-9(piperudin-4-yl)-7,9-dihydro-8H-purin- 8-one (568 mg, 1.46 mmol) in MeOH (6 ml). The reaction mixture was stirred at room temperature for 30 minutes. Sodium cyanotrihydroborate (275 mg, 4.38 mmol) was added at room temperature, then the reaction mixture was heated at 80°C for 30 minutes, then allowed to cool to room temperature and concentrated in vacuo. The resulting crude product was purified by fcc, eluting with a gradient of 0-10% MeOH in (0.5% NH3 in DCM) to give the title compound (390 mg, 66%) as a white solid.

1H NMR (400 MHz, DMSO) 1.67-1.71 (2H, m), 1.99-2.08 (2H, m), 2.25 (3H, s), 2.54 (3H, s ), 2.60-2.74 (2H, m), 2.92 (2H, d), 3.34 (3H, s), 4.12-4.22 (1H, m), 7.34- 7.42 (1H, m), 7.84 (1H, s), 8.18 (1H, s), 8.28-8.30 (1H, m), 8.50 (1H, s), 8 .58 (1H, s), 8.70-8.72 (1H, m);

mass/charge MN ⁺ 404.

Form A.

The final product, 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8H-purin-8-one, was analyzed by XRPD and DSC and found it to be crystalline. Using XRPD of a sample of the material, the diffraction pattern shown in FIG. 1. Form A of 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin-8one is characterized by at least with one peak at a 2θ value of 7.1 or 8.5°, as measured using CuKa radiation. The ten most pronounced XRPD peaks are shown in Table. IN.

- 79 043669

Table B

Ten most pronounced XRPD peaks for Form A of 7-methyl-9-(1methylpiperidin-4-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin-8-one

Angle 2-theta (2Θ) Intensity, % 8.5 100.0 7.1 51.2 18.8 47.1 21.5 44.8 15.4 39.1 26.2 37.5 16.3 29.7 14.2 21.0 12.7 17.8 19.8 17.4

where 2-theta values are ±0.2°.

Example 45: 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro8H-purin-8-one.

RuPhos Pd G3 (47.5 mg, 0.06 mmol) and RuPhos (26.5 mg, 0.06 mmol) were added to 2-chloro-7-methyl-9-(1methylpiperidin-4-yl)-7.9 -dihydro-8H-purin-8-one (80 mg, 0.28 mmol), 7-methylquinoxaline-6-amine (54.2 mg, 0.34 mmol) and CS ₂ CO ₃ (185 mg, 0.57 mmol) in 1,4-dioxane (6 ml) at room temperature. The reaction mixture was heated at 100°C for 4 hours, then allowed to cool to room temperature, concentrated in vacuo and the residue purified by fcc, elution gradient from 0 to 10% MeOH in DCM, then further purified by preparative HPLC to give the title compound (49 mg, 43%) as a yellow solid.

^1H NMR (400 MHz, DMSO) 1.68 (2H, d), 1.97 (2H, t), 2.17 (3H, s), 2.52-2.63 (5H, m), 2 .88 (2H, d), 3.34 (3H, s), 4.11-4.25 (1H, m), 7.90 (1H, s), 8.20 (1H, s), 8, 62 (2H, d), 8.73 (1H, d), 8.79 (1H, d);

mass/charge MN ⁺ 405.

Example 46: 9-((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)7,9-dihydro-8H -purin-8-one.

Sodium triacetoxyborohydride (118 mg, 0.55 mmol) was added to a suspension of formaldehyde (37% solution in water, 90 mg, 1.11 mmol) and 9-((3S,4R)-3-fluoropiperidin-4-yl)-7 -methyl-2-((7-methylquinolin-6yl)amino)-7,9-dihydro-8H-purin-8-one (113 mg, 0.28 mmol) in iPrOH (2 ml) at room temperature. Add MeOH (1 ml) and the reaction mixture was stirred at room temperature for 30 min. Sodium triacetoxyborohydride (59 mg, 0.27 mmol) was added and the reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was quenched using sat. aq. NaHCO ₃ solution (2 ml) and extracted with EtOAc (2x2 ml). The combined organic layers were washed with sat. saline solution (1 ml), passed through filter paper to separate the phases and concentrated in vacuo. MeCN (3 mL) was added to the resulting solid, and the solid was isolated by filtration to obtain the title compound (25 mg, 21%) as a yellow solid.

1H NMR (400 MHz, DMSO) 1.03-1.11 (1H, m), 1.83 (1H, s), 2.29 (3H, s), 2.53 (3H, s), 2, 89 (1H, s), 3.18 (1H, s), 3.36 (3H, s), 3.44-3.57 (1H, m), 4.25-4.41 (1H, m) , 4.56-4.72 (1H, m), 4.85 (1H, d), 7.40 (1H, dd), 7.83

- 80 043669 (1H, s), 8.21 (1H, s), 8.25 (1H, d), 8.46 (1H, s), 8.58 (1H, s), 8.71 (1H ,dd);

mass/charge MN ⁺ 422.

Example 47: 7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(1-(oxetan-3-yl)piperidin-4-yl)-7,9dihydro-8H-purine-8 -He.

Oxetan-3-one (30.5 mg, 0.42 mmol) was added to

7-methyl-2-((7-methylquinolin-6-yl)amino)-9(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one (150 mg, 0.39 mmol) in MeOH (5 ml). The reaction mixture was heated at 60°C for 1 hour. NaBH ₃ CN (36.3 mg, 0.58 mmol) was added and the reaction mixture was heated at 60°C for 16 hours, then allowed to cool to room temperature, and was also poured into water (125 ml) and extracted with DCM (3x125 ml). The combined organic layers were dried over Na ₂ SO ₄ , filtered and evaporated, then purified by preparative HPLC to give the title compound (21 mg, 12%) as a white solid.

^1H NMR (400 MHz, DMSO) 1.91-2.08 (2H, d), 2.33-2.53 (2H, t), 2.56 (3H, d), 2.67-2, 71 (2H, m), 2.89 (2H, d), 3.33 (3H, s), 3.44-3.47 (1H, m), 4.22-4.23 (1H, m) , 4.48-4.59 (2H, t), 4.61-4.62 (2H, t), 7.42-7.45 (1H, m), 7.85 (1H, s), 8 .21 (1H, s), 8.48 (1H, s), 8.52-8.54 (1H, m), 8.71-8.75 (2H, m);

mass/charge MN ⁺ 446.

Example 48: 9-(1-(2-hydroxyethyl)piperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8H-purin-8-one .

2-Bromoethan-1-ol (52.9 mg, 0.42 mmol) was added to

7-methyl-2-((7-methylquinolin-6-yl)amino)-9(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one (150 mg, 0.39 mmol) and DIPEA (0.202 ml, 1.16 mmol) in iPrOH (3 ml). The reaction mixture was heated at 80°C for 16 hours, allowed to cool to room temperature and concentrated in vacuo. The resulting crude product was purified by preparative HPLC to obtain the title compound (18 mg, 11%) as a white solid.

1H NMR (300 MHz, CD3OD) 1.81-1.85 (2H, m), 2.25-2.59 (2H, m), 2.59-2.65 (5H, m), 2.74 -2.88 (2H, m), 3.16-3.20 (2H, m), 3.42 (3H, s), 3.73 (2H, t), 4.31-4.41 (1H , m), 7.43-7.47 (1H, m), 7.87 (1H, s), 8.09 (1H, s), 8.34-8.37 (1H, m), 8, 67-8.69 (2H, m);

mass/charge MN ⁺ 434.

Example 49: 9-(1-(2-methoxyethyl)piperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8H-purin-8-one .

1-Bromo-2-methoxyethane (58.9 mg, 0.42 mmol) was added to 7-methyl-2-((7-methylquinolin-6-yl)amino)9-(piperidin-4-yl)-7, 9-dihydro-8H-purin-8-one (150 mg, 0.39 mmol) and DIPEA (0.202 ml, 1.16 mmol) in iPrOH (5 ml). The reaction mixture was heated at 80°C for 16 hours. The reaction mixture was allowed to cool to room temperature and directly purified by preparative HPLC to obtain the title compound (26 mg, 15%) as a yellow solid.

- 81 043669

1H NMR (300 MHz, CDCI3) 1.69 (2H, d), 2.27 (2H, s), 2.64 (3H, s), 2.73 (2H, s), 2.92 (2H, s), 3.19 (2H,

d), 3.21-3.63 (6H, m), 3.63-3.68 (2H, m), 4.40 (1H, t), 7.15 (1H, s), 7.28 -7.34 (1H, m), 7.96 (2H, d), 8.30 (1H,

d), 8.76-8.78 (1H, m), 8.97 (1H, s);

mass/charge MN ⁺ 448.

Example 50: 9-(1-ethylpinerudin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one.

Acetaldehyde (18.7 mg, 0.42 mmol) was added to 7-methyl-2-((7-methylquinolin-6-yl)amino)-9(piperidin-4-yl)-7,9-dihydro-8H- purin-8-one (150 mg, 0.39 mmol) in MeOH (5 ml). The reaction mixture was stirred at RT for 1 hour. NaBH ₃ CN (36.9 mg, 0.58 mmol) was added and the reaction mixture was stirred at RT for 1 hour, then poured into water (150 ml) and extracted with DCM (3 x 150 ml). The combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo, then purified by preparative HPLC to give the title compound (49 mg, 31%) as a yellow solid.

1H NMR (300 MHz, CDCl ₃ ) 1.22 (3H, s), 1.85-1.88 (2H, d), 2.15 (2H, s), 2.57-2.63 (5H, s), 2.91 (2H, s), 3.20 (2H, s), 3.46 (3H, s), 4.41-4.45 (1H, m), 7.16 (1H, s ), 7.28-7.33 (1H, m), 7.95 (2H, d), 8.35-8.37 (1H, m), 8.77 (1H, s), 9.01 ( 1H, s);

mass/charge MN ⁺ 418.

Example 51: 9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H purin-8-one.

Cesium carbonate (3.09 g, 9.49 mmol) was added to 9-(1-acetylpiperidin-4-yl)-2-chloro-7-methyl-7,9dihydro-8H-purin-8-one (840 mg, 2.71 mmol) and 7-methylquinolin-6-amine hydrochloride (528 mg, 2.71 mmol) in 1,4-dioxane (15 ml). Brettphos Pd G3 (123 mg, 0.14 mmol) was added and the reaction mixture was heated at 100°C for 1 hour, then allowed to cool to room temperature, filtered and washed with DCM (10 ml). The combined filtrate was concentrated in vacuo and the residue was purified by fcc, eluent with 0-10% MeOH in DCM. The resulting oil was taken up in MeCN (20 ml) and the resulting suspension was briefly refluxed before being allowed to cool to room temperature. The resulting precipitate was isolated by filtration, washed with a small amount of MeCN and dried in vacuo to give the title compound ( 550 mg, 47.0%) as a pale yellow solid.

1H NMR (400 MHz, DMSO) 1.81 (2H, t), 1.93 (3H, s), 2.21 -2.32 (1H, m), 2.38-2.47 (1H, m ), 2.52 (3H, s), 2.62 (1H, t), 3.15 (1H, t), 3.34 (3H, s), 3.94 (1H, d), 4.44 (1H, ddd), 4.57 (1H, d), 7.39 (1H, dd), 7.85 (1H, s), 8.13 (1H, d), 8.17 (1H, s) , 8.29 (1H, s), 8.59 (1H, s), 8.74 (1H, dd);

mass/charge MN ⁺ 432.

Example 52: 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9dihydro-8H-purine-8- He.

M HCl in 1,4-dioxane (11.40 mL, 45.59 mmol) was added in one portion to the stirred solution of tert-butyl-(3R,4R)-3-fluoro-4-(7-methyl-2-(( 7-methylquinolin-6-yl)amino)-8-oxo-7,8-dihydro-9H-82 043669 purin-9-yl)pyrrolidin-1-carboxylate (1.5 g, 3.04 mmol) in 1, 4-dioxane (50 ml) at room temperature and the reaction mixture was stirred at room temperature for 12 hours, then concentrated in vacuo. The resulting residue was diluted with DCM (200 ml), then made basic with 7 M NH ₃ in MeOH (10 ml). The mixture was washed with sat. saline solution, dried over Na ₂ SO ₄ , filtered and concentrated in vacuo. The resulting crude product was purified by preparative HPLC (XBridge Prep C18 OBD column, 5 µm silica, 19 mm diameter, 150 mm length) using mixtures of decreasing polarity water (containing 0.1% NH4HCO3) and MeCN as eluents, then triturated with MeCN to produce one batch of the title compound. The liquids obtained after trituration were purified by preparative HPLC (column: XBridge Shield RP18 OBD column, 19x250 mm, 10 µm; mobile phase A: water (10 mmol/l NH4HCO3), mobile phase B: ACN; flow rate: 25 ml/min; gradient: 25% B to 35% B in 7 min; 254; 220 nm; retention time: 6.83 min) to give additional title compound. The two batches were combined to give the title compound (620 mg, 52%) as a white solid.

¹ H NMR (300 MHz, DMSO, 23°C) 2.49 (3H, s), 2.80-3.02 (2H, m), 3.02-3.31 (2H, m), 3, 34 (3H, s), 4.75 (1H, dt), 5.57 (1H, dd), 7.41 (1H, dd), 7.86 (1H, s), 8.12-8.25 (3H, m), 8.67 (1H, s), 8.74 (1H, dd), one NH missing;

mass/charge MH+394.

Form A.

Final product, 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)7,9-dihydro-8H-purine-8 -one was analyzed by XRPD and found to be crystalline. XRPD of a sample of the material produced the diffraction pattern shown in FIG. 5. Form A 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8Hpurin-8 -it is characterized by at least one peak at a 2θ value of 7.3° or 15.0°, as measured using CuKa radiation. The ten most pronounced XRPD peaks are shown in Table. WITH.

Table C

Ten most prominent XRPD peaks for Form A 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2((7-methylquinolin-6-yl)amino)-7,9-dihydro- 8H-purin-8-one, ranked by intensity, with the first value being the highest intensity

where 2-theta values are ±0.2°.

Bibliography.

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Buisson R et al. Distinct but concerted roles of ATR, DNA-PK and Chkl in countering replication stress during S phase. Molecular Cell 2015; 59:1011-1024

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Claims (13)

Medunjanin S et al. Interaction of the double strand break repair kinase DNA-PK and estrogen receptor alpha. Mol Biol Cell 2010; 21: 1620-1628 Munck JM et al. Chemosensitization of cancer cells by KU-0060648, a dual inhibitor of DNA-PK and PI-3K. Mol Cancer Ther 2012; 11: 1789-1798 Neal JA and Meek K. Choosing the right path: does DNA-PK help make the decision? Mutat Res 2011; 711:73-86 Riabinska A et al. Therapeutic targeting of a robust non-oncogene addiction to PRKDC in ATM-defective tumors. Science Translational Medicine 2013; 189: 189ra78 San Filippo J et al. Mechanism of ukaryotic homologous recombination. Annu Rev Biochem 2008; 77: 229-257 Smith GCM and Jackson SP. The DNA dependent protein kinase. Genes and Development 1999;13: 916-934 Symington LS and Gautier J. Double strand break end resection and repair pathway choice. Annu Rev Genet 2011; 45: 247-271 Willmore E et al. A novel DNA-dependent protein kinase inhibitor, NU7026, potentiates the cytotoxicity of topoisomerase II poisons used in the treatment of leukemia Blood 2004; 103:4659-4665 Yoo S and Dynan WS. Geometry of a complex formed by double strand break repair proteins at a single DNA end: recruitment of DNA-PKcs induces inward translocation of Ku protein. Nucleic Acids Res 1999; 27: 4679-4 CLAIM 1. Compound of formula (I) (I), or a pharmaceutically acceptable salt thereof, wherein A ¹ is N or CR ^2A , A ² is N or CR ^2B and A ³ is N or CR ^2C , wherein not more than one of A ¹ , A ² and A ³ represents N; R1 is cyclohexanyl, piperidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, oxetanyl and pyrrolidinyl, where ^R1 is optionally substituted with one or more groups selected from fluorine, methyl, ethyl, hydroxyl, NH ₂ , dioxo, C(O)Me and oxetanyl, wherein ethyl is optionally substituted with hydroxyl or methoxy; R ^2A , R ^2b and R ^2C are each independently hydrogen, methyl or methoxy. 2. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein A ¹ is CR ^2A , A ² is CR ^2B and A ³ is CR ^2C . 3. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any one of the preceding - 85 043669 points, where each of R ^2A , R ^2B and R ^2C represents hydrogen. 4. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of the preceding claims, wherein R ¹ is selected from piperidinyl and pyrrolidinyl. 5. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of the preceding claims, wherein R ¹ is optionally substituted with one or more groups selected from fluorine, methyl, ethyl, hydroxyl, NH ₂ and oxetanyl. 6. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of the preceding claims, wherein R ¹ is pyrrolidin-3-yl. 7. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to any of the preceding claims, wherein R ¹ is 4-fluoropyrrolidin-3-yl. 8. A compound of formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is selected from the group consisting of 9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-7,9-dihydro-8H-purin-8one; 9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro-8H-purin-8one; 9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinazolin-6-yl)amino)-7,9-dihydro-8H-purin-8one; 9-(1-acetylpiperidin-4-yl)-2-((2,7-dimethylquinoxalin-6-yl)amino)-7-methyl-7,9-dihydro-8Hpurin-8-one; 9-(1-acetylpiperidin-4-yl)-2-((3,7-dimethylquinoxalin-6-yl)amino)-7-methyl-7,9-dihydro-8Hpurin-8-one; 9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin8-one; 9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one; 2-((4,7-dimethylquinolin-6-yl)amino)-9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl-7,9-dihydro-8Hpurin-8-one; 9-((1r,4r)-4-hydroxycyclohexyl)-2-((4-methoxy-7-methylquinolin-6-yl)amino)-7-methyl-7,9-dihydro8H-purin-8-one; 9-((1r,4r)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one; 9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin8-one; 9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one; 9-((1s,4s)-4-hydroxycyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one; 9-((1s,4s)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro8H-purin-8-one; 9-((1r,4r)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro8H-purin-8-one; 9-((1s,4s)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-7,9-dihydro8H-purin-8-one; 9-((1r,4r)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-7,9-dihydro8H-purin-8-one; 9-((1r,4r)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro8H-purin-8-one; 9-((1s,4s)-4-hydroxy-4-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro8H-purin-8-one; 9-((1s,4s)-4-hydroxy-1-methylcyclohexyl)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro8H-purin-8-one; (S)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8one; (S)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin8-one; (R)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin-8one; (R)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydrofuran-3-yl)-7,9-dihydro-8H-purin8-one; (R)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin8-one; - 86 043669 (R)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8Hpurin-8- she; (S)-7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8H-purin8-one; (S)-7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2H-pyran-3-yl)-7,9-dihydro-8Hpurin-8-one; 7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8one; 7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8one; 7-methyl-2-((7-methylquinoxalin-6-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin8-one; 7-methyl-2-((7-methylquinazolin-6-yl)amino)-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8H-purin-8one; 2-((2,7-dimethylquinoxalin-6-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8Hpurin-8-one; 2-((3,7-dimethylquinoxalin-6-yl)amino)-7-methyl-9-(tetrahydro-2H-pyran-4-yl)-7,9-dihydro-8Hpurin-8-one; 9-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro8H-purin-8-one; 7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(oxetan-3-yl)-7,9-dihydro-8H-purin-8-one; 7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(piperidin-4-yl)-7,9-dihydro-8H-purin-8-one; 9-((3S,4R)-3-fluoropiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin8-one; 9-((1s,4s)-4-amino-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one; 9-((1g,4g)-4-amino-4-methylcyclohexyl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one; (R)-7-methyl-9-(1-methylpyrrolidin-3-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin8-one; (S)-7-methyl-9-(1-methylpyrrolidin-3-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin-8one; 7-methyl-2-((7-methylcinnolin-6-yl)amino)-9-(1-methylpiperidin-4-yl)-7,9-dihydro-8H-purin-8-one; 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin-8-one; 7-methyl-9-(1-methylpiperidin-4-yl)-2-((7-methylquinoxalin-6-yl)amino)-7,9-dihydro-8H-purin-8one; 9-((3S,4R)-3-fluoro-1-methylpiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro8H-purine-8 -she; 7-methyl-2-((7-methylquinolin-6-yl)amino)-9-(1-(oxetan-3-yl)piperidin-4-yl)-7,9-dihydro-8Hpurin-8-one; 9-(1-(2-hydroxyethyl)piperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one; 9-(1-(2-methoxyethyl)piperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one; 9-(1-ethylpiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin-8-one; 9-(1-acetylpiperidin-4-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8H-purin-8-one; and 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino)-7,9-dihydro-8Hpurin-8-one. 9. A compound of formula (I) according to claim 1, where the compound is 9-((3R,4R)-4fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino) -7,9-dihydro-8H-purin-8-one or a pharmaceutically acceptable salt thereof. 10. The compound of formula (I) according to claim 1, where the compound is 9-((3R,4R)-4fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinolin-6-yl)amino) -7,9-dihydro-8H-purin-8-one. 11. The compound of formula (I) according to claim 1, where the compound is the crystalline form of 9-((3R,4R)-4-fluoropyrrolidin-3-yl)-7-methyl-2-((7-methylquinoline-6- yl)amino)-7,9-dihydro-8H-purin-8one and is characterized by X-ray powder diffraction (XRPD) peaks at 2 theta (2θ) values of 7.3° plus/minus 0.2°, 14.6° plus/minus 0.2° and 15.0° plus/minus 0.2°, as measured using CuKa radiation. 12. The compound of formula (I) according to claim 11, wherein the compound is characterized by an X-ray powder pattern (XRPD) substantially as shown in FIG. 5 as measured using CuKa radiation. 13. The compound of formula (I) according to claim 1, where the compound is a crystalline form -