WO2019002442A1 - Chemical compounds - Google Patents

Abstract

The present invention relates to compounds of formula (I) and pharmaceutically acceptable salts thereof, processes and intermediates used for their preparation, pharmaceutical compositions containing them and their use in the treatment of cellular proliferative disorders.

Description

CHEMICAL COMPOUNDS

This specification relates to certain indole compounds and pharmaceutically acceptable salts thereof that selectively down-regulate the estrogen receptor and possess anti-cancer activity. This specification also relates to use of said indole compounds and pharmaceutically acceptable salts thereof in methods of treatment of the human or animal body, for example in prevention or treatment of cancer. This specification also relates to processes and intermediate compounds involved in the preparation of said indole compounds and to pharmaceutical compositions containing them.

Estrogen receptor alpha (ERa, ESR1, NR3A) and estrogen receptor beta (ER , ESR2, NR3b) are steroid hormone receptors which are members of the large nuclear receptor family. Structured similarly to all nuclear receptors, ERa is composed of six functional domains (named A-F) (Dahlman- Wright, et al., Pharmacol. Rev., 2006, 58:773- 781) and is classified as a ligand-dependent transcription factor because after its association with the specific ligand, (the female sex steroid hormone 17b estradiol (E2)), the complex binds to genomic sequences, named Estrogen Receptor Elements (ERE) and interacts with co-regulators to modulate the transcription of target genes. The ERa gene is located on 6q25.1 and encodes a 595 AA protein and multiple isoforms can be produced due to alternative splicing and translational start sites. In addition to the DNA binding domain (Domain C) and the ligand binding domain (Domain E) the receptor contains a N- terminal (A/B) domain, a hinge (D) domain that links the C and E domains and a C- terminal extension (F domain). While the C and E domains of ERa and ER are quite conserved (96% and 55%> amino acid identity respectively) conservation of the A/B, D and F domains is poor (below 30% amino acid identity). Both receptors are involved in the regulation and development of the female reproductive tract and in addition play roles in the central nervous system, cardiovascular system and in bone metabolism. The genomic action of ERs occurs in the nucleus of the cell when the receptor binds EREs directly (direct activation or classical pathway) or indirectly (indirect activation or non-classical pathway). In the absence of ligand, ERs are associated with heat shock proteins, Hsp90 and Hsp70, and the associated chaperone machinery stabilizes the ligand binding domain (LBD) making it accessible to ligand. Liganded ER dissociates from the heat shock proteins leading to a conformational change in the receptor that allows dimerisation, DNA binding, interaction with co-activators or co-repressors and modulation of target gene expression. In the non-classical pathway, AP-1 and Sp-1 are alternative regulatory DNA sequences used by both isoforms of the receptor to modulate gene expression. In this example, ER does not interact directly with DNA but through associations with other DNA bound transcription factors e.g. c-Jun or c-Fos (Kushner et al, Pure Applied Chemistry 2003, 75: 1757-1769). The precise mechanism whereby ER affects gene transcription is poorly understood but appears to be mediated by numerous nuclear factors that are recruited by the DNA bound receptor. The recruitment of co-regulators is primarily mediated by two protein surfaces, AF2 and AF1 which are located in E-domain and the A/B domain respectively. AF1 is regulated by growth factors and its activity depends on the cellular and promoter environment whereas AF2 is entirely dependent on ligand binding for activity. Although the two domains can act independently, maximal ER transcriptional activity is achieved through synergistic interactions via the two domains (Tzukerman, et al, Mol. Endocrinology, 1994, 8:21-30). Although ERs are considered transcription factors they can also act through non-genomic mechanisms as evidenced by rapid ER effects in tissues following E2 administration in a timescale that is considered too fast for a genomic action. It is still unclear if receptors responsible for the rapid actions of estrogen are the same nuclear ERs or distinct G-protein coupled steroid receptors (Warner, et al, Steroids 2006 71:91-95) but an increasing number of E2 induced pathways have been identified e.g. MAPK/ER pathway and activation of endothelial nitric oxide synthase and PI3K/Akt pathway. In addition to ligand dependent pathways, ERa has been shown to have ligand independent activity through AF-1 which has been associated with stimulation of MAPK through growth factor signalling e.g. insulin like growth factor 1 (IGF-1) and epidermal growth factor (EGF). Activity of AF-1 is dependent on

phosphorylation of Serl 18 and an example of cross-talk between ER and growth factor signalling is the phosphorylation of Ser 118 by MAPK in response to growth factors such as IGF-1 and EGF (Kato, et al, Science, 1995, 270: 1491-1494).

A large number of structurally distinct compounds have been shown to bind to ER. Some compounds such as endogenous ligand E2, act as receptor agonists whereas others competitively inhibit E2 binding and act as receptor antagonists. These compounds can be divided into 2 classes depending on their functional effects. Selective estrogen receptor modulators (SERMs) such as tamoxifen have the ability to act as both receptor agonists and antagonists depending on the cellular and promoter context as well as the ER isoform targeted. For example tamoxifen acts as an antagonist in breast but acts as a partial agonist in bone, the cardiovascular system and uterus. All SERMs appear to act as AF2 antagonists and derive their partial agonist characteristics through AFl . A second group, fulvestrant being an example, are classified as full antagonists and are capable of blocking estrogen activity via the complete inhibition of AFl and AF2 domains through induction of a unique conformation change in the ligand binding domain (LBD) on compound binding which results in complete abrogation of the interaction between helix 12 and the remainder of the LBD, blocking co-factor recruitment (Wakeling, et al, Cancer Res., 1991, 51 :3867-3873; Pike, et al, Structure, 2001, 9: 145-153).

Intracellular levels of ERa are down-regulated in the presence of E2 through the ubiquitin/proteosome (Ub/26S) pathway. Polyubiquitinylation of liganded ERa is catalysed by at least three enzymes; the ubiquitin-activating enzyme El activated ubiquitin is conjugated by E2 with lysine residues through an isopeptide bond by E3 ubiquitin ligase and polyubiquitinated ERa is then directed to the proteosome for degradation. Although ER-dependent transcription regulation and proteosome -mediated degradation of ER are linked (Lonard, et al, Mol. Cell, 2000 5:939-948), transcription in itself is not required for ERa degradation and assembly of the transcription initiation complex is sufficient to target ERa for nuclear proteosomal degradation. This E2 induced degradation process is believed to necessary for its ability to rapidly activate transcription in response to requirements for cell proliferation, differentiation and metabolism (Stenoien, et al., Mol. Cell Biol., 2001, 21 :4404-4412). Fulvestrant is also classified as a selective estrogen receptor down- regulator (SERD), a subset of antagonists that can also induce rapid down-regulation of ERa via the 26S proteosomal pathway. In contrast a SERM such as tamoxifen can increase ERa levels although the effect on transcription is similar to that seen for a SERD.

Approximately 70% of breast cancers express ER and/or progesterone receptors implying the hormone dependence of these tumour cells for growth. Other cancers such as ovarian and endometrial are also thought to be dependent on ERa signalling for growth. Therapies for such patients can inhibit ER signalling either by antagonising ligand binding to ER e.g. tamoxifen which is used to treat early and advanced ER positive breast cancer in both pre and post menopausal setting; antagonising and down-regulating ERa e.g. fulvestrant which is used to treat breast cancer in women which have progressed despite therapy with tamoxifen or aromatase inhibitors; or blocking estrogen synthesis e.g.

aromatase inhibitors which are used to treat early and advanced ER positive breast cancer. Although these therapies have had an enormously positive impact on breast cancer treatment, a considerable number of patients whose tumours express ER display de novo resistance to existing ER therapies or develop resistance to these therapies over time. Several distinct mechanisms have been described to explain resistance to first-time tamoxifen therapy which mainly involve the switch from tamoxifen acting as an antagonist to an agonist, either through the lower affinity of certain co-factors binding to the tamoxifen-ERa complex being off-set by over-expression of these co-factors, or through the formation of secondary sites that facilitate the interaction of the tamoxifen-ERa complex with co-factors that normally do not bind to the complex. Resistance could therefore arise as a result of the outgrowth of cells expressing specific co-factors that drive the tamoxifen-ERa activity. There is also the possibility that other growth factor signalling pathways directly activate the ER receptor or co-activators to drive cell proliferation independently of ligand signalling.

More recently, mutations in ESR1 have been identified as a possible resistance mechanism in metastatic ER-positive patient derived tumour samples and patient-derived xenograft models (PDX) at frequencies varying from 17-25%. These mutations are predominantly, but not exclusively, in the ligand-binding domain leading to mutated functional proteins; examples of the amino acid changes include Ser463Pro, Val543Glu, Leu536Arg, Tyr537Ser, Tyr537Asn and Asp538Gly, with changes at amino acid 537 and 538 constituting the majority of the changes currently described. These mutations have been undetected previously in the genomes from primary breast samples characterised in the Cancer Genome Atlas database. Of 390 primary breast cancer samples positive for ER expression not a single mutation was detected in ESR1 (Cancer Genome Atlas Network, 2012 Nature 490: 61-70). The ligand binding domain mutations are thought to have developed as a resistance response to aromatase inhibitor endocrine therapies as these mutant receptors show basal transcriptional activity in the absence of estradiol. The crystal structure of ER, mutated at amino acids 537 and 538, showed that both mutants favoured the agonist conformation of ER by shifting the position of helix 12 to allow co-activator recruitment and thereby mimicking agonist activated wild type ER. Published data has shown that endocrine therapies such as tamoxifen and fulvestrant can still bind to ER mutant and inhibit transcriptional activation to some extent and that fulvestrant is capable of degrading Try537Ser but that higher doses may be needed for full receptor inhibition (Toy et al, Nat. Genetics 2013. 45: 1439-1445; Robinson et al, Nat. Genetics 2013, 45: 144601451; Li, S. et al. Cell Rep. 4, 1116-1130 (2013). It is therefore feasible that certain compounds of the Formula (I) or pharmaceutically acceptable salts thereof (as described hereinafter) will be capable of down-regulating and antagonising mutant ER although it is not known at this stage whether ESR1 mutations are associated with an altered clinical outcome.

Regardless of which resistance mechanism or combination of mechanisms takes place, many are still reliant on ER-dependent activities and removal of the receptor through a SERD mechanism offers the best way of removing the ERa receptor from the cell.

Fulvestrant is currently the only SERD approved for clinical use, yet despite its

mechanistic properties, the pharmacological properties of the drug have limited its efficacy due to the current limitation of a 500mg monthly dose which results in less than 50% turnover of the receptor in patient samples compared to the complete down-regulation of the receptor seen in in vitro breast cell line experiments (Wardell, et al., Biochem. Pharm., 2011, 82:122-130). Hence there is a need for new ER targeting agents that have the required pharmaceutical properties and SERD mechanism to provide enhanced benefit in the early, metastatic and acquired resistance setting.

The compounds of the specification have been found to possess potent anti-tumour activity, being useful in inhibiting the uncontrolled cellular proliferation which arises from malignant disease. The compounds of the specification provide an anti-tumour effect by, as a minimum, acting as SERDs. For example, the compounds of the specification may exhibit anti -tumour activity via the ability to down-regulate the estrogen receptor in a number of different breast cancer cell-lines, for example against the MCF-7, CAMA-1, BT474 and/or MDA-MB-134 breast cancer cell-lines. Such compounds may be expected to be more suitable as therapeutic agents, particularly for the treatment of cancer.

The compounds of the specification may also exhibit advantageous physical properties (for example, lower lipophilicity, higher aqueous solubility, higher permeability, lower plasma protein binding, and/or greater chemical stability), and/or favourable toxicity profiles (for example a decreased activity at hERG), and/or favourable metabolic or pharmacokinetic profiles, in comparison with other known SERDs. Such compounds may therefore be especially suitable as therapeutic agents, particularly for the treatment of cancer.

According to one aspect of the specification there is provided a compound of Formula (I):

wherein:

A is CR⁶ or N;

R¹ is H, F or OMe;

R³ is H, F, Me or CH₂OH;

R⁴ is F or Me;

R⁵ is H or F; and

R⁶ is H or F;

or a pharmaceutically acceptable salt thereof.

This specification also describes pharmaceutical compositions which comprise a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable excipient.

This specification also describes a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use as a medicament. This specification also describes a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer.

This specification also describes combinations of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, with another anti-tumour agent, for use in the treatment of cancer.

Further aspects of the specification will be apparent to one skilled in the art from reading this specification.

In one embodiment there is provided a compound of Formula (I) as defined above.

In one embodiment there is provided a pharmaceutically acceptable salt of a compound of Formula (I).

In a further embodiment A is CR⁶.

In one embodiment A is N.

In one embodiment A is CH.

In one embodiment R¹ is F.

In one embodiment R¹ is OMe.

In one embodiment R¹ is H.

In one embodiment A is N and R¹ is H.

In one embodiment A is CH and R¹ is OMe.

In one embodiment ²

In one embodiment ³

In one embodiment ³

In one embodiment ³

In one embodiment

In one embodiment

In one embodiment

In one embodiment

or a pharmaceutically acceptable salt thereof.

In one embodiment A is N; R¹ is H; R³ is F or Me; R⁴ is F or Me; and R⁵ is H; or a pharmaceutically acceptable salt thereof. In one embodiment A is N; R¹ is H; R³ is F; R⁴ is F; and R⁵ is H; or a

pharmaceutically acceptable salt thereof.

In one embodiment A is N; R¹ is H; R³ is Me; R⁴ is Me; and R⁵ is H; or a pharmaceutically acceptable salt thereof.

In one embodiment A is CH; R¹ is F or OMe; R³ is F or Me; R⁴ is F or Me; and R⁵ is H; or a pharmaceutically acceptable salt thereof.

In one embodiment A is CH; R¹ is OMe; R³ is F; R⁴ is F; and R⁵ is H; or a pharmaceutically acceptable salt thereof.

In one embodiment A is CH; R¹ is OMe; R³ is Me; R⁴ is Me; and R⁵ is H; or a pharmaceutically acceptable salt thereof.

In a further aspect there is provided a compound of Formula (IZ):

(IZ)

wherein:

A is CR⁶ or N;

R¹ is H F or OMe;

R³ is H, F, Me or CH₂OH;

R⁴ is F or Me;

R⁵ is H or F; and

R⁶ is H or F: or a pharmaceutically acceptable salt thereof.

In a further embodiment there is provided the compound of Formula (IZ) or a pharmaceutically acceptable salt thereof wherein the stereochemistry at the 1 -position of the tetrahydro-3H-pyrrolo[3,2-f]isoquinoline ring is S.

In a further embodiment there is provided the compound of Formula (IZ) or a pharmaceutically acceptable salt thereof wherein the stereochemistry at the 1 -position of the tetrahydro-3H-pyrrolo[3,2-f]isoquinoline ring is R.

In a further embodiment there is provided the compound of Formula (IZ) or a pharmaceutically acceptable salt thereof wherein the stereochemistry at the 3 -position of the tetrahydro-3H-pyrrolo[3,2-f]isoquinoline ring is S.

In a further embodiment there is provided the compound of Formula (IZ) or a pharmaceutically acceptable salt thereof wherein the stereochemistry at the 3 -position of the tetrahydro-3H-pyrrolo[3,2-f]isoquinoline ring is R.

In one embodiment there is provided a compound of Formula (I), wherein the compound is selected from the group consisting of:

N-(l-(3-fiuoropropyl)azetidin-3-yl)-6-((6S,8R)-8-methyl-7-(2,2,2-trifiuoroethyl)-6,7,8,9- tetrahydro-3H-pyrrolo[3,2-fJisoquinolin-6-yl)pyridin-3-amine; and

(6S,8R)-7-(2-fluoro-2-methylpropyl)-6-(4-(2-(3-(fluoromethyl)azetidin-l-yl)ethoxy)-2- methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrrolo[3,2-f]isoquinoline;

or a pharmaceutically acceptable salt thereof.

Reference herein to compounds of Formula (I) is to be understood as referring to compounds of Formula (I) and/or (IZ) unless stated otherwise.

In one embodiment there is provided a compound of Formula (I), or a

pharmaceutically acceptable salt thereof, wherein the compound is selected from any of the Examples in the specification. A further feature is any of the embodiments described in the specification with the proviso that any of the specific Examples are individually disclaimed. A further feature is any of the embodiments described in the specification with the proviso that any one or more of the compounds selected from the above list of examples of compounds of the specification are individually disclaimed.

The compounds of Formula (I) have two or more chiral centres and it will be recognised that the compounds of Formula (I) may be prepared, isolated and/or supplied with or without the presence, in addition, of one or more of the other possible enantiomeric and/or diastereomeric isomers of the compounds of Formula (I) in any relative proportions. The preparation of enantioenriched/ enantiopure and/or diastereoenriched/ diastereopure compounds may be carried out by standard techniques of organic chemistry that are well known in the art, for example by synthesis from enantioenriched or enantiopure starting materials, use of an appropriate enantioenriched or enantiopure catalyst during synthesis, and/or by resolution of a racemic or partially enriched mixture of stereoisomers, for example via chiral chromatography.

For use in a pharmaceutical context it may be preferable to provide a compound of Formula (I) or a pharmaceutically acceptable salt thereof without large amounts of the other stereoisomeric forms being present.

Accordingly, in one embodiment there is provided a composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, optionally together with one or more of the other stereoisomeric forms of the compound of Formula (I), or pharmaceutically acceptable salt thereof, wherein the compound of Formula (I), or pharmaceutically acceptable salt thereof is present within the composition with a diastereomeric excess (%de) of > 90%.

In a further embodiment the %de in the above-mentioned composition is > 95%.

In a further embodiment the %de in the above-mentioned composition is > 98%.

In a further embodiment the %de in the above-mentioned composition is > 99%.

In a further embodiment there is provided a composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, optionally together with one or more of the other stereoisomeric forms of the compound of Formula (I), or

pharmaceutically acceptable salt thereof, wherein the compound of Formula (I), or pharmaceutically acceptable salt thereof is present within the composition with an enantiomeric excess (%>ee) of > 90%>.

In a further embodiment the %ee in the above-mentioned composition is > 95%.

In a further embodiment the %ee in the above-mentioned composition is > 98%.

In a further embodiment the %ee in the above-mentioned composition is > 99%.

In a further embodiment there is provided a composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, optionally together with one or more of the other stereoisomeric forms of the compound of Formula (I), or pharmaceutically acceptable salt thereof, wherein the compound of Formula (I), or pharmaceutically acceptable salt thereof is present within the composition with an enantiomeric excess (%ee) of > 90% and a diastereomeric excess (%de) of > 90%.

In further embodiments of the above-mentioned composition the %ee and %de may take any combination of values as listed below:

• The %ee is <5% and the %de is≥ 80%.

• The %ee is <5% and the %de is≥ 90%.

• The %ee is <5% and the %de is≥ 95%.

• The %ee is <5% and the %de is≥ 98%.

• The %ee is≥ 95% and the %de is≥ 95%.

• The %ee is≥ 98% and the %de is≥ 98%.

• The %ee is≥ 99% and the %de is≥ 99%.

In a further embodiment there is provided a pharmaceutical composition which comprises a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable excipient.

In one embodiment there is provided a pharmaceutical composition which comprises a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable excipient, optionally further comprising one or more of the other stereoisomeric forms of the compound of (I), or pharmaceutically acceptable salt thereof, wherein the compound of Formula (I) or pharmaceutically acceptable salt thereof is present within the composition with an enantiomeric excess (%ee) of > 90%.

In a further embodiment the %ee in the above-mentioned composition is > 95%.

In a further embodiment the %ee in the above-mentioned composition is > 98%.

In a further embodiment the %ee in the above-mentioned composition is > 99%.

In one embodiment there is provided a pharmaceutical composition which comprises a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable excipient, optionally further comprising one or more of the other stereoisomeric forms of the compound of Formula (I), or pharmaceutically acceptable salt thereof, wherein the compound of Formula (I), or pharmaceutically acceptable salt thereof is present within the composition with a diastereomeric excess (%de) of > 90%.

In a further embodiment the %de in the above-mentioned composition is > 95%.

In a further embodiment the %de in the above-mentioned composition is > 98%.

In a further embodiment the %de in the above-mentioned composition is > 99%.

In one embodiment there is provided a pharmaceutical composition which comprises a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in association with a pharmaceutically acceptable excipient, optionally further comprising one or more of the other stereoisomeric forms of the compound of Formula (I), or pharmaceutically acceptable salt thereof, wherein the compound of Formula (I), or pharmaceutically acceptable salt thereof is present within the composition with an enantiomeric excess (%>ee) of > 90%> and a diastereomeric excess (%>de) of > 90%>.

In further embodiments of the above-mentioned pharmaceutical composition the %ee and %de may take any combination of values as listed below:

• The %ee is≥ 95% and the %de is≥ 95%.

• The %ee is≥ 98% and the %de is≥ 98%.

• The %ee is≥ 99% and the %de is≥ 99%.

The compounds of Formula (I), and pharmaceutically acceptable salts thereof may be prepared, used or supplied in amorphous form, crystalline form, or semicrystalline form and any given compound of Formula (I), or pharmaceutically acceptable salt thereof may be capable of being formed into more than one crystalline / polymorphic form, including hydrated (e.g. hemi-hydrate, a mono-hydrate, a di-hydrate, a tri-hydrate or other stoichiometry of hydrate) and/or solvated forms. It is to be understood that the present specification encompasses any and all such solid forms of the compound of Formula (I), and pharmaceutically acceptable salts thereof.

In further embodiments there is provided a compound of Formula (I), which is obtainable by the methods described in the 'Examples' section hereinafter.

The present specification is intended to include all isotopes of atoms occurring in the present compounds. Isotopes will be understood to include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. Isotopes of carbon include C and C. Isotopes of nitrogen include ¹⁵N.

A suitable pharmaceutically acceptable salt of a compound of the Formula (I), is, for example, an acid addition salt. A suitable pharmaceutically acceptable salt of a compound of Formula (I), may be, for example, an acid-addition salt of a compound of the Formula (I).

A further suitable pharmaceutically acceptable salt of a compound of the Formula (I), is, for example, a salt formed within the human or animal body after administration of a compound of the Formula (I), to said human or animal body.

The compound of Formula (I), or pharmaceutically acceptable salt thereof may be prepared as a co-crystal solid form. It is to be understood that a pharmaceutically acceptable co-crystal of a compound of the Formula (I), or pharmaceutically acceptable salts thereof, form an aspect of the present specification.

For the avoidance of doubt it is to be understood that where in this specification a group is qualified by 'hereinbefore defined' or 'defined herein' the said group

encompasses the first occurring and broadest definition as well as each and all of the alternative definitions for that group.

For the further avoidance of doubt, the group denotes the point of attachment to the remainder of the compound of Formula (I).

Another aspect of the present specification provides a process for preparing a compound of the Formula (I), or a pharmaceutically acceptable salt thereof. A suitable process is illustrated by the following representative process variants in which, unless otherwise stated, A and R¹ to R⁵ have any of the meanings defined hereinbefore.

Necessary starting materials may be obtained by standard procedures of organic chemistry. The preparation of such starting materials is described in conjunction with the following representative process variants and within the accompanying Examples. Alternatively, necessary starting materials are obtainable by analogous procedures to those illustrated which are within the ordinary skill of an organic chemist.

Compounds of Formula (I), illustrated here wherein R⁵ is hydrogen, may be prepared by, for example:

a) etherification of a suitable aryl or heteroaryl halide of Formula (II), bromide shown although other halides and activating groups may be appropriate, with an alcohol corresponding to R² using a suitable metal catalyst (for example RockPhos 3rd Generation Precatalyst) in a suitable solvent (such as toluene or DME) in the presence of a suitable base (such as cesium carbonate) and a suitable temperature (such as 90-120 °C).

b) amination of a suitable aryl or heteroaryl halide of Formula (II) bromide shown although other halides and activating groups may be appropriate, with an amine corresponding to R² using a suitable metal catalyst (for example BrettPhos or RuPhos, and Pd₂(dba)3) in a suitable solvent (for example 1 ,4-dioxane) in the presence of a suitable base (for example cesium carbonate, sodium tert-butoxide, or LiHMDS) at a suitable temperature (such as 90-130 °C).

c) alkylation of a suitable phenol or hydroxyl heteroaryl compound of Formula (III) with an alcohol corresponding to R² via Mitsunobu reaction using appropriate reagents (such as triphenylphosphine and diisopropyl (E)-diazene-l ,2-dicarboxylate) in a suitable solvent (such as THF); removal of the protecting group (PG), such as Boc, in Formula (III), using acidic conditions (for example anhydrous HC1 in a methanol) at suitable temperature (such as 10-30 °C).

d) Alkylation of amines of Formula (IV) with a suitable alkyl halide (such as bromide as shown although other halides such as iodide or chloride and other leaving groups may be appropriate) of Formula (V) in a suitable solvent (such as acetonitrile) in the presence of a suitable base (such as potassium carbonate) at a suitable temperature (such as 80-90 °C).

e) alkylation of a suitable amine of Formula (VI) with alkylhalides (such as iodide as shown although other halides such as bromide or chloride and other leaving groups may be appropriate) of Formula VII) in a suitable solvent (such as acetonitrile) in the presence of a suitable base (such as potassium carbonate) and a suitable temperature (such as 80-90°C).

(VI) (VII)

Compounds of Formula (II) may be prepared by a number of methods, for example: a) cyclization reactions of aniline compounds of Formula (VIII) in the presence of pentamethylcyclopentadienyl iridium dichloride with a suitable base (such as potassium carbonate) in a suitable solvent (such as toluene) after stirring at a suitable temperate (such as 90-110 °C) for a suitable duration (such as 10-30 hours).

(VIII) b) alkylation of a compound of Formula (IX) with a compound of Formula (X), wherein LG is a suitable leaving group, for example a halogen atom (such as bromo or chloro) or trifluoromethanesulfonate, in the presence of a suitable base (such as diisopropylethylamine) in a suitable solvent (for example DCM or 1 ,4-dioxane) and at a suitable temperature (such as 20- 85°C).

(IX) (X)

Compounds of Formula (III) may be prepared from compounds of Formula (II) through a reaction sequence involving protection of the indole nitrogen with a suitable group (such as Boc) and conversion of the halide to a hydroxyl group in the presence of an alkaline solution (such as aqueous potassium hydroxide), a suitable palladium catalyst (such as Pd₂dba3), and a suitable ligand (such as di-tert-butyl(2',4',6'-triisopropyl-3,4,5,6- tetramethyl-[l,l'-biphenyl]-2-yl)phosphane) in a suitable solvent (such as 1 ,4-dioxane) at a suitable temperature (such as 80- 110 °C). Compounds of Formula (IV) may be prepared from a compound of Formula (II) under conditions known in the art as suitable for Mitsunobu reactions using appropriate reagents (such as triphenylphosphine and diisopropyl (E)-diazene-l ,2-dicarboxylate) with 2-haloethanol (such as 2-bromoethan-l-ol) in a suitable solvent (such as THF).

Compounds of Formula (VI) may be prepared from aryl halides of Formula (II) with tert-butyl 3-aminoazetidine-l-carboxylate using a suitable metal catalyst (such as for example RuPhos or BrettPhos and Pd₂(dba)3) in a suitable solvent (for example 1 ,4- dioxane) in the presence of a suitable base (for example cesium carbonate, sodium tert- butoxide, or LiHMDS) at a suitable temperature (such as 90-130 °C); the Boc protecting group may be subsequently removed using an acid (such as trifluoro acetic acid) in a suitable solvent (such as DCM).

Compounds of Formula (VIII) may be prepared by reaction of a compound of Formula (XI) with a aldehyde compound of Formula (XII), under conditions known in the art as suitable for Pictet-Spengler reactions, such as in the presence of a suitable Lewis acid (such as Ytterbium(III) trifluoromethanesulfonate) and in a suitable solvent (for example acetonitrile) and at a suitable temperature (such as 50-80 °C), followed by the removal of the protecting group (such as 4-MeO-benzyl) in the presence of a suitable acidic condition (such has anhydrous hydrochloride in methanol) at a suitable temperature (such as 10 -30

^•C). r

(XI) (XII)

Compounds of Formula (XI) may be prepared by, for example:

a) reaction of a compound of Formula (XIII) with an aldehyde of Formula (XIV), in a suitable solvent (for example THF) in the presence of a suitable reducing agent (such as sodium triacetoxyborohydride) and at a suitable temperature (such as 20-30°C).

(XIII) (XIV) (XV)

b) reaction of a compound of Formula (XIII) with a compound of Formula (X), wherein LG is a suitable leaving group (for example a halogen atom (such as bromo or chloro) or trifluoromethanesulfone), in the presence of a suitable base (such as

diisopropylethylamine) in a suitable solvent (for example DCM or 1,4-dioxane) and at a suitable temperature (such as 20-85°C).

c) reaction of a compound of Formula (XIII) with an acid of Formula (XV) under standard amide bond forming conditions (for example in the presence of an amide coupling reagent (such as HATU) and a suitable base (such as triethylamine) in a suitable solvent (such as DMF)), followed by reduction of the resultant amide bond using a suitable reducing agent (such as borane) in a suitable solvent (such as THF) at a suitable temperature (such as 60-70°C).

Compounds of Formula (XIII) may be prepared by a number of methods known to the art for the synthesis of chiral amines, for example, via ring opening of sulfamidates of Formula (XIV) according to the scheme shown below.

(XIV) (XV) (XI II)

Step 1 : Alkynation, e.g. n-butyllithium/THF/-78 °C to room temperature

Step 2: Amine deprotection, e.g. anhydrous HCI/MeOH/1 ,4-dioxane

Step 3: Aniline deprotection, e.g. NH₂OH«HCI/ethanol/reflux to remove 4-MeO-benzyl

Compounds of Formula (IX) may be prepared from compounds of Formula (XVI) using a suitable reducing reagent (such as lithium aluminum hydride) in the presence of a suitable chelating Lewis acid (such as trimethylaluminum) in a suitable solvent (such as THF) at a suitable temperature (such as -78 °C to room temperature). Diastereomeric and enantiomeric mixtures of the reduction reaction may be separated using suitable chiral chromatography methods to provide enantiomeric pure compounds of Formula (IX). r

(XVI)

Compounds of Formula (XVI) may be prepared by reaction of a compound of Formula (XVII) via a sequence of reactions: (i) amide formation of amine compounds of Formula (XVI) and carboxylic acid compounds of Formula (XVIII) using a suitable coupling reagent (such as HATU) in the presence of a suitable base (such as triethylamine) in a suitable solvent (such as DMF); (ii) cyclization under conditions known in the art as suitable for Bischler-Napieralski reaction, such as heating under microwave in the presence of phosphorus chrolide at a suitable temperature (such as 100-120 °C).

I

(XVII) (XVIII)

Compounds of Formula (XVII) may be prepared by a number of methods known to the art for the synthesis of secondary amines, for example, via reductive amination of 1- (lH-indol-4-yl)propan-2-one using a suitable amination reagent (such as ammonium formate) and using a suitable metal calalyst (such as palladium on carbon) in a suitable solvent (such as methanol). It is to be understood that other permutations of the process steps in the process variants described above are also possible.

When a pharmaceutically acceptable salt of a compound of Formula (I) is required it may be obtained by, for example, reaction of said compound with a suitable acid or suitable base.

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

A suitable protecting group for an amino or alkylamino group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or t-butoxycarbonyl group, an

arylmethoxycarbonyl group, for example benzyloxycarbonyl, or an aroyl group, for example benzoyl. The deprotection conditions for the above protecting groups necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or alkoxycarbonyl group or an aroyl group may be removed for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide. Alternatively an alkoxycarbonyl group such as a t-butoxycarbonyl group may be removed, for example, by treatment with a suitable acid as hydrochloric, sulphuric, formic, phosphoric or trifluoroacetic acid, and an arylmethoxycarbonyl group such as a benzyloxycarbonyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon, or by treatment with a Lewis acid, such as boron tris(trifluoroacetate). A suitable alternative protecting group for a primary amino group is, for example, a phthaloyl group, which may be removed by treatment with an alkylamine, for example dimethylaminopropylamine, or hydrazine.

A suitable protecting group for a hydroxy group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an aroyl group, for example benzoyl, an arylmethyl group, for example benzyl, or a trialkyl or diarylalkyl silane, such as TBDMS or TBDPS. The deprotection conditions for the above protecting groups will necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or an aroyl group may be removed, for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide.

Alternatively an arylmethyl group such as a benzyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.

A suitable protecting group for a carboxy group is, for example, an esterifying group, for example a methyl or an ethyl group which may be removed, for example, by hydrolysis with a base such as sodium hydroxide, or for example a t-butyl group which may be removed, for example, by treatment with an acid, such as trifluoroacetic acid, or for example a benzyl group which may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.

The protecting groups may be removed at any convenient stage in the synthesis using conventional techniques well known in the chemical art.

Certain of the intermediates defined herein are novel and these are provided as further features of the specification.

Biological Assays

The following assays were used to measure the effects of the compounds of the present specification.

ERq binding assay

The ability of compounds to bind to isolated Estrogen Receptor Alpha Ligand binding domain (ER alpha - LBD (GST)) was assessed in competition assays using a LanthaScreen™ Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) detection end-point. For the LanthaScreen TR-FRET endpoint, a suitable fluorophore (Fluormone ES2, ThermoFisher, Product code P2645) and recombinant human Estrogen Receptor alpha ligand binding domain, residues 307-554 (expressed and purified in-house) were used to measure compound binding. The assay principle is that ER alpha -LBD (GST) is added to a fluorescent ligand to form a receptor/fluorophore complex. A terbium- labelled anti-GST antibody (Product code PV3551) is used to indirectly label the receptor by binding to its GST tag, and competitive binding is detected by a test compound's ability to displace the fluorescent ligand, resulting in a loss of TR-FRET signal between the Tb- anti-GST antibody and the tracer. The assay was performed as follows with all reagent additions carried out using the Beckman Coulter BioRAPTR FRD microfluidic workstation:

1. Acoustic dispense 120 nL of the test compound into a black low volume 384 well assay plates.

2. Prepare lx ER alpha -LBD/Tb-antiGST Ab in ES2 screening buffer and incubate for 15 minutes.

3. Dispense 6 of the lx AR-LBD/Tb-anti-GST Ab reagent into each well of the assay plate followed by 6 of Fluorophore reagent into each well of the assay plate

4. Cover the assay plate to protect the reagents from light and evaporation, and incubate at room temperature for 4 hours.

5. Excite at 337 nm and measure the fluorescent emission signal of each well at 490 nm and 520 nm using the BMG PheraSTAR.

Compounds were dosed directly from a compound source microplate containing serially diluted compound (4 wells containing 10 mM, 0.1 mM, 1 mM and 10 nM final compound respectively) to an assay microplate using the Labcyte Echo 550. The Echo 550 is a liquid handler that uses acoustic technology to perform direct microplate -to-microplate transfers of DMSO compound solutions and the system can be programmed to transfer multiple small nL volumes of compound from the different source plate wells to give the desired serial dilution of compound in the assay which is then back-filled to normalise the DMSO concentration across the dilution range.

In total 120 nL of compound plus DMSO were added to each well and compounds were tested in a 12-point concentration response format over a final compound

concentration range of 10, 2.917, 1.042, 0.2083, 0.1, 0.0292, 0.0104, 0.002083, 0.001, 0.0002917, 0.0001042, and 0.00001 μΜ respectively. TR-FRET dose response data obtained with each compound was exported into a suitable software package (such as Origin or Genedata) to perform curve fitting analysis. Competitive ER alpha binding was expressed as an IC50 value. This was determined by calculation of the concentration of compound that was required to give a 50% reduction in tracer compound binding to ER alpha-LBD.

MCF-7 ER downregulation assay The ability of compounds to down-regulate Estrogen Receptor (ER) numbers was assessed in a cell based immuno-fluorescence assay using the MCF-7 human ductal carcinoma breast cell line. MCF-7 cells were revived directly from a cryovial (approx 5 x 10⁶ cells) in Assay Medium (phenol red free Dulbecco's Modified Eagle's medium

(DMEM); Sigma D5921) containing 2mM L-Glutamine and 5% (v/v) Charcoal/Dextran treated foetal calf serum. Cells were syringed once using a sterile 18G x 1.5 inch (1.2 x 40 mm) broad gauge needle and cell density was measured using a Coulter Counter

(Beckman). Cells were further diluted in Assay Medium to a density of 3.75 x 10⁴ cells per mL and 40 per well added to transparent bottomed, black, tissue culture -treated 384 well plates (Costar, No. 3712) using a Thermo Scientific Matrix WellMate or Thermo Multidrop. Following cell seeding, plates were incubated overnight at 37 °C, 5% C0₂ (Liconic carousel incubator). Test data was generated using the LabCyte Echo™ model 555 compound reformatter which is part of an automated workcell (Integrated Echo 2 workcell). Compound stock solutions (10 mM) of the test compounds were used to generate a 384 well compound dosing plate (Labcyte P-05525-CV1). 40 of each of the 10 mM compound stock solutions was dispensed into the first quadrant well and then 1 : 100 step-wise serial dilutions in DMSO were performed using a Hydra II (MATRIX UK) liquid handling unit to give 40 of diluted compound into quadrant wells 2 (0.1 mM), 3 (1 μΜ) and 4 (0.01 μΜ), respectively. 40 μΕ of DMSO added to wells in row P on the source plate allowed for DMSO normalisation across the dose range. To dose the control wells 40 μΕ of DMSO was added to row 01 and 40 μΕ of 100 μΜ fulvestrant in DMSO was added to row 03 on the compound source plate.

The Echo uses acoustic technology to perform direct microplate-to-microplate transfers of DMSO compound solutions to assay plates. The system can be programmed to transfer volumes as low as 2.5 nL in multiple increments between microplates and in so doing generates a serial dilution of compound in the assay plate which is then back-filled to normalise the DMSO concentration across the dilution range. Compounds were dispensed onto the cell plates with a compound source plate prepared as above producing a 12 point duplicate 3 μΜ to 3 pM dose range with 3 -fold dilutions and one final 10-fold dilution using the Integrated Echo 2 workcell. The maximum signal control wells were dosed with DMSO to give a final concentration of 0.3%, and the minimum signal control wells were dosed with fulvestrant to give a final concentration of 100 nM accordingly. Plates were further incubated for 18-22 hours at 37 °C, 5% C0₂ and then fixed by the addition of 20 μΐ, of 11.1% (v/v) formaldehyde solution (in phosphate buffered saline (PBS)) giving a final formaldehyde concentration of 3.7% (v/v). Cells were fixed at room temperature for 20 mins before being washed two times with 250 μΐ_^ PBS/Proclin (PBS with a Biocide preservative) using a BioTek platewasher, 40 μΐ_^ of PBS/Proclin was then added to all wells and the plates stored at 4 °C. The fixing method described above was carried out on the Integrated Echo 2 workcell. Immunostaining was performed using an automated AutoElisa workcell. The PBS/Proclin was aspirated from all wells and the cells permeabilised with 40 PBS containing 0.5% Tween™ 20 (v/v) for 1 hour at room temperature. The plates were washed three times in 250 of PBS/0.05%> (v/v) Tween 20 with Proclin (PBST with a Biocide preservative) and then 20 μΐ_^ of ERa (SP1) Rabbit monoclonal antibody (Thermofisher) 1 : 1000 in PBS/Tween™/3% (w/v) Bovine Serum Albumin was added. The plates were incubated overnight at 4 °C (Liconic carousel incubator) and then washed three times in 250 μΐ_^ of PBS/0.05% (v/v) Tween™ 20 with Proclin (PBST). The plates were then incubated with 20 μΕΛνεΙΙ of a goat anti-rabbit IgG AlexaFluor 594 or goat anti-rabbit AlexaFluor 488 antibody (Molecular Probes) with Hoechst at 1 :5000 in PBS/Tween™/3% (w/v) Bovine Serum Albumin for lhour at room temperature. The plates were then washed three times in 250 μΐ, of PBS/0.05%> (v/v) Tween™ 20 with Proclin (PBST with a Biocide preservative). 20 μΐ, of PBS was added to each well and the plates covered with a black plate seal and stored at 4 °C before being read. Plates were read using a Cellomics Arrayscan reading the 594 nm (24 hr time point) or 488 nm (5 hr timepoint) fluorescence to measure the ERa receptor level in each well. The mean total intensity was normalized for cell number giving the total intensity per cell. The data was exported into a suitable software package (such as Origin) to perform curve fitting analysis. Down-regulation of the ERa receptor was expressed as an IC50 value and was determined by calculation of the concentration of compound that was required to give a 50% reduction of the average maximum Total Intensity signal.

The data shown in Table A were generated (the data below may be a result from a single experiment or an average of two or more experiments): Table A

According to a further aspect of the specification there is provided a pharmaceutical composition, which comprises a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in association with a pharmaceutically acceptable excipient.

Suitable pharmaceutically acceptable excipients for a tablet formulation include, for example, inert diluents, granulating and disintegrating agents, binding agents, lubricating agents, preservative agents and antioxidants. A further suitable pharmaceutically acceptable excipient may be a chelating agent. Tablet formulations may be uncoated or coated either to modify their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, in either case, using conventional coating agents and procedures well known in the art.

Compositions for oral use may alternatively be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil.

Aqueous suspensions generally contain the active ingredient in finely powdered form together with one or more suspending agents, dispersing or wetting agents. The aqueous suspensions may also contain one or more preservatives, anti-oxidants, colouring agents, flavouring agents, and/or sweetening agents.

Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil or in a mineral oil. The oily suspensions may also contain a thickening agent. Sweetening agents such as those set out above, and flavouring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water generally contain the active ingredient together with a dispersing or wetting agent, suspending agent and one or more preservatives. Additional excipients such as sweetening, flavouring and colouring agents, may also be present.

The pharmaceutical compositions of the specification may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil or a mineral oil or a mixture of any of these. The emulsions may also contain sweetening, flavouring and preservative agents.

Syrups and elixirs may be formulated with sweetening agents, and may also contain a demulcent, preservative, flavouring and/or colouring agent.

For further information on formulation the reader is referred to Chapter 25.2 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.

The amount of active ingredient that is combined with one or more excipients to produce a single dosage form will necessarily vary depending upon the host treated and the particular route of administration. For example, oral administration to humans will generally require, for example, from 1 mg to 2 g of active agent to be administered compounded with an appropriate and convenient amount of excipients which may vary from about 3 to about 98 percent by weight of the total composition. It will be understood that, if a large dosage is required, multiple dosage forms may be required, for example two or more tablets or capsules, with the dose of active ingredient divided conveniently between them. Typically, unit dosage forms will contain about 10 mg to lg of a compound of this specification.

The size of the dose for therapeutic or prophylactic purposes of compounds of the present specification will naturally vary according to the nature and severity of the disease state, the age and sex of the animal or patient and the route of administration, according to well known principles of medicine.

In using compounds of the present specification for therapeutic or prophylactic purposes it will generally be administered so that a daily dose in the range, for example, 1 mg/kg to 100 mg/kg body weight is received, given if required in divided doses.

In one aspect of the specification, compounds of the present specification or pharmaceutically acceptable salts thereof, are administered as tablets comprising lOmg to lOOOmg of the compound of the specification (or a pharmaceutically acceptable salt thereof), wherein one or more tablets are administered as required to achieve the desired dose.

As stated above, it is known that signalling through ERa causes tumourigenesis by one or more of the effects of mediating proliferation of cancer and other cells, mediating angiogenic events and mediating the motility, migration and invasiveness of cancer cells. We have found that the compounds of the present specification possess potent anti-tumour activity which it is believed is obtained by way of antagonism and down-regulation of ERa that is involved in the signal transduction steps which lead to the proliferation and survival of tumour cells and the invasiveness and migratory ability of metastasising tumour cells.

Accordingly, the compounds of the present specification may be of value as anti- tumour agents, in particular as selective inhibitors of the proliferation, survival, motility, dissemination and invasiveness of mammalian cancer cells leading to inhibition of tumour growth and survival and to inhibition of metastatic tumour growth. Particularly, the compounds of the present specification may be of value as anti-proliferative and anti- invasive agents in the containment and/or treatment of solid tumour disease. Particularly, the compounds of the present specification may be useful in the prevention or treatment of those tumours which are sensitive to inhibition of ERa and that are involved in the signal transduction steps which lead to the proliferation and survival of tumour cells and the migratory ability and invasiveness of metastasising tumour cells. Further, the compounds of the present specification may be useful in the prevention or treatment of those tumours which are mediated alone or in part by antagonism and down-regulation of ERa, i.e. the compounds may be used to produce an ERa inhibitory effect in a warm-blooded animal in need of such treatment.

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore for use as a medicament.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, as a medicament.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament. According to a further aspect of the specification, there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore for use in the production of an anti-proliferative effect in a warm-blooded animal such as man.

According to a further aspect of the specification, there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for the production of an anti-proliferative effect in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in the production of an antiproliferative effect in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided a method for producing an anti-proliferative effect in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore for use in a warm-blooded animal such as man as an anti-invasive agent in the containment and/or treatment of solid tumour disease.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, as an anti-invasive agent in the containment and/or treatment of solid tumour disease.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in a warm-blooded animal such as man as an anti-invasive agent in the containment and/or treatment of solid tumour disease.

According to a further aspect of the specification there is provided a method for producing an anti-invasive effect by the containment and/or treatment of solid tumour disease in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification, there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for use in the prevention or treatment of cancer in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in the prevention or treatment of cancer in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in the manufacture of a medicament for use in the prevention or treatment of cancer in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided a method for the prevention or treatment of cancer in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification, there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore for use in the prevention or treatment of solid tumour disease in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the prevention or treatment of solid tumour disease in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in the prevention or treatment of solid tumour disease in a warm-blooded animal such as man.

According to a further aspect of the specification there is provided a method for the prevention or treatment of solid tumour disease in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for use in the prevention or treatment of those tumours which are sensitive to inhibition of ERa that are involved in the signal transduction steps which lead to the proliferation, survival, invasiveness and migratory ability of tumour cells.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the prevention or treatment of those tumours which are sensitive to inhibition of ERa that are involved in the signal transduction steps which lead to the proliferation, survival, invasiveness and migratory ability of tumour cells.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in the prevention or treatment of those tumours which are sensitive to inhibition of ERa that are involved in the signal transduction steps which lead to the proliferation, survival, invasiveness and migratory ability of tumour cells.

According to a further aspect of the specification there is provided a method for the prevention or treatment of those tumours which are sensitive to inhibition of ERa that are involved in the signal transduction steps which lead to the proliferation, survival, invasiveness and migratory ability of tumour cells which comprises administering to said animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore for use in providing an inhibitory effect on ERa.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in providing an inhibitory effect on ERa. According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in the manufacture of a medicament for use in providing an inhibitory effect on ERa.

According to a further aspect of the specification there is provided a method for providing an inhibitory effect on ERa which comprises administering an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for use in providing a selective inhibitory effect on ERa.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in providing a selective inhibitory effect on ERa.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in providing a selective inhibitory effect on ERa.

According to a further aspect of the specification there is also provided a method for providing a selective inhibitory effect on ERa which comprises administering an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

Described herein are compounds that can bind to ERa ligand binding domain and are selective estrogen receptor degraders. In biochemical and cell based assays the compounds of the present specification are shown to be potent estrogen receptor binders and reduce cellular levels of ERa and may therefore be useful in the treatment of estrogen sensitive diseases or conditions (including diseases that have developed resistance to endocrine therapies), i.e. for use in the treatment of cancer of the breast and gynaecological cancers (including endometrial, ovarian and cervical) and cancers expressing ERa mutated proteins which may be de novo mutations or have arisen as a result of treatment with a prior endocrine therapy such as an aromatase inhibitor. According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for use in the treatment of breast or gynaecological cancers.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the treatment of breast or gynaecological cancers.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in the treatment of breast or gynaecological cancers.

According to a further aspect of the specification there is provided a method for treating breast or gynaecological cancers, which comprises administering an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for use in the treatment of cancer of the breast, endometrium, ovary or cervix.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the treatment of cancer of the breast, endometrium, ovary or cervix.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in the treatment of cancer of the breast, endometrium, ovary or cervix.

According to a further aspect of the specification there is provided a method for treating cancer of the breast, endometrium, ovary or cervix, which comprises administering an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for use in the treatment of breast cancer. According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the treatment of breast cancer.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in the treatment of breast cancer.

According to a further aspect of the specification there is provided a method for treating breast cancer, which comprises administering an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for use in the treatment of breast cancer, wherein the cancer has developed resistance to one or more other endocrine therapies.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the treatment of breast cancer, wherein the cancer has developed resistance to one or more other endocrine therapies.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, in the manufacture of a medicament for use in the treatment of breast cancer, wherein the cancer has developed resistance to one or more other endocrine therapies.

According to a further aspect of the specification there is provided a method for treating breast cancer, wherein the cancer has developed resistance to one or more other endocrine therapies, which comprises administering an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

In one feature of the specification, the cancer to be treated is breast cancer. In a further aspect of this feature, the breast cancer is Estrogen Receptor +ve (ER+ve).

According to a further aspect of the specification there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore, for use in the treatment of ER+ve breast cancer. According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined herein before in the treatment of ER+ve breast cancer.

According to a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined herein before in the manufacture of a medicament for use in the treatment of ER+ve breast cancer.

According to a further aspect of the specification there is provided a method for treating ER+ve breast cancer, which comprises administering an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore.

The anti-cancer treatment defined herein may be applied as a sole therapy or may involve, in addition to the compounds of the specification, conventional surgery or radiotherapy or chemotherapy.

Accordingly, in one embodiment there is provided a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and an additional anti-tumour substance for the conjoint treatment of cancer.

In a further embodiment there is provided the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in combination with an additional anti-tumour substance for the conjoint treatment of cancer.

In a further embodiment there is provided the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in combination with an additional anti-tumour substance for manufacture of a medicament for the conjoint treatment of cancer.

In one embodiment there is provided a method of the conjoint treatment of cancer in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and an additional anti-tumour substance.

In a further embodiment there is provided a combination suitable for use in the treatment of cancer comprising a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, and another anti-tumour agent, wherein the another anti -tumour agent is the standard of care for the specific cancer to be treated; the person skilled in the art will understand the meaning of "standard of care". In a further embodiment there is provided the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in combination with another anti-tumour agent, for the treatment of cancer, wherein the another anti -tumour agent is the standard of care for the specific cancer to be treated; the person skilled in the art will understand the meaning of "standard of care".

In a further embodiment there is provided the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in combination with an another anti-tumour agent, for manufacture of a medicament for the treatment of cancer, wherein the another anti-tumour agent is the standard of care for the specific cancer to be treated; the person skilled in the art will understand the meaning of "standard of care".

In one embodiment there is provided a method of treating cancer in a warmblooded animal, such as man, in need of such treatment which comprises administering to said animal a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and another anti-tumour agent, wherein the another anti-tumour agent is the standard of care for the specific cancer to be treated; the person skilled in the art will understand the meaning of "standard of care".

In a further embodiment there is provided a combination suitable for use in the treatment of cancer comprising a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, and an antihormonal agent, such as an antioestrogen (for example tamoxifen, fulverstrant, toremifene, raloxifene, droloxifene and iodoxyfene), a

progestrogen (for example megestrol acetate) or an aromatase inhibitor (for example anastrozole, letrozole, vorazole and exemestane).

In a further embodiment there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in combination with and an antihormonal agent, such as an antioestrogen (for example tamoxifen, fulverstrant, toremifene, raloxifene, droloxifene and iodoxyfene), a progestrogen (for example megestrol acetate) or an aromatase inhibitor (for example anastrozole, letrozole, vorazole and exemestane) for the treatment of cancer.

In a further embodiment there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in combination with and an antihormonal agent, such as an antioestrogen (for example tamoxifen, fulverstrant, toremifene, raloxifene, droloxifene and iodoxyfene), a progestrogen (for example megestrol acetate) or an aromatase inhibitor (for example anastrozole, letrozole, vorazole and exemestane) in the manufacture of a medicament for the treatment of cancer.

In a further embodiment there is provided a method of treating cancer in a warmblooded animal, such as man, in need of such treatment which comprises administering to said animal a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and an antihormonal agent, such as an antioestrogen (for example tamoxifen, fulverstrant, toremifene, raloxifene, droloxifene and iodoxyfene), a progestrogen (for example megestrol acetate) or an aromatase inhibitor (for example anastrozole, letrozole, vorazole and exemestane).

In a further aspect of the specification there is provided a combination suitable for use in the treatment of cancer comprising a compound of the Formula (I), or a

pharmaceutically acceptable salt thereof, and an mTOR inhibitor, such as AZD2014.

In a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in combination with an mTOR inhibitor, such as AZD2014, for the treatment of cancer.

In a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in combination with an mTOR inhibitor, such as AZD2014, in the manufacture of a medicament for the treatment of cancer.

In a further aspect of the specification there is provided a method of treating cancer in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and an mTOR inhibitor, such as AZD2014.

In a further aspect of the specification there is provided a combination suitable for use in the treatment of cancer comprising a compound of the Formula (I), or a

pharmaceutically acceptable salt thereof, and a CDK4/6 inhibitor, such as palbociclib.

In a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in combination with a CDK4/6 inhibitor, such as palbociclib, for the treatment of cancer.

In a further aspect of the specification there is provided the use of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in combination with a CDK4/6 inhibitor, such as palbociclib, in the manufacture of a medicament for the treatment of cancer.

In a further aspect of the specification there is provided a method of treating cancer in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and a CDK4/6 inhibitor, such as palbociclib.

In one aspect the above combinations of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, with another anti-tumour agent, is suitable for use in the treatment of breast or gynaecological cancers, such as cancer of the breast, endometrium, ovary or cervix, particularly breast cancer, such as ER+ve breast cancer.

Herein, where the term "combination" is used it is to be understood that this refers to simultaneous, separate or sequential administration. In one aspect of the specification "combination" refers to simultaneous administration. In another aspect of the specification "combination" refers to separate administration. In a further aspect of the specification "combination" refers to sequential administration. Where the administration is sequential or separate, the delay in administering the second component should not be such as to lose the beneficial effect of the combination. Where a combination of two or more components is administered separately or sequential, it will be understood that the dosage regime for each component may be different to and independent of the other components.

Conveniently, the compounds of the present specification are dosed once daily.

According to a further aspect of the present specification there is provided a kit comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof in combination with another anti-tumour agent selected from those described herein.

According to a further aspect of the present specification there is provided a kit comprising:

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

b) another anti-tumour agent selected from those described herein herein above in a second unit dosage form; and

c) container means for containing said first and second dosage forms.

According to a further aspect of the present specification there is provided a kit comprising: a) a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, in a first unit dosage form;

b) an anti-tumour agent selected from an anti-hormonal agent, an mTOR inhibitor, or a CDK4/6 inhibitor, in a second unit dosage form; and

c) container means for containing said first and second dosage forms.

Combination therapy as described above may be added on top of standard of care therapy typically carried out according to its usual prescribing schedule.

Although the compounds of the Formula (I), are primarily of value as therapeutic agents for use in warm-blooded animals (including man), they are also useful whenever it is required to inhibit ER-a. Thus, they are useful as pharmacological standards for use in the development of new biological tests and in the search for new pharmacological agents.

Personalised Healthcare

Another aspect of the present specification is based on identifying a link between the status of the gene encoding ERa and potential susceptibility to treatment with a compound of Formula (I). In particular, ERa gene status may indicate that a patient is less likely to respond to exisiting hormone therapy (such as aromatase inhibitors), in part at least because some ERa mutations are though to arise as resistance mechanisms to existing treatments. A SERD, particularly a SERD which can be administered orally in potentially larger doses without excessive inconvenince, may then advantageously be used to treat patients with ERa mutations who may be resistant to other therapies. This therefore provides opportunities, methods and tools for selecting patients for treatment with a compound of Formula (I), particularly cancer patients. The present specification relates to patient selection tools and methods (including personalised medicine). The selection is based on whether the tumour cells to be treated possess wild-type or mutant ERa gene. The ERa gene status could therefore be used as a biomarker to indicate that selecting treatment with a SERD may be advantageous. For the avoidance of doubt, compounds of the Formula (I), as described herein, are thought to be similarly active against wild-type and mutant ERa genes, at least those mutations in ERa gene identified at the date of filing this application.

There is a clear need for biomarkers that will enrich for or select patients whose tumours will respond to treatment with a SERD, such as a compound of Formula (I). Patient selection biomarkers that identify the patients most likely to respond to one agent over another are ideal in the treatment of cancer, since they reduce the unnecessary treatment of patients with non-responding tumours to the potential side effects of such agents.

Tumours which contain wild type ERa are believed to be susceptible to treatment with a compound of Formula (I), for example as a first-line treatment. Tumours may also respond to treatment with a compound of Formula (I), as a second- line, third-line or subsequent therapy and this may be useful, in particular, where the tumours contain mutant ERa and may thus be resistant to existing therapies such as AIs.

For the purpose of this specification, a gene status of wild-type is meant to indicate normal or appropriate expression of the gene and normal function of the encoded protein. In contrast, mutant status is meant to indicate expression of a protein with altered function, consistent with the known roles of mutant ERa genes in cancer. Any number of genetic or epigenetic alterations, including but not limited to mutation, amplification, deletion, genomic rearrangement, or changes in methylation profile, may result in a mutant status. However, if such alterations nevertheless result in appropriate expression of the normal protein, or a functionally equivalent variant, then the gene status is regarded as wild-type. Examples of variants that typically would not result in a functional mutant gene status include synonymous coding variants and common polymorphisms (synonymous or non- synonymous). As discussed below, gene status can be assessed by a functional assay, or it may be inferred from the nature of detected deviations from a reference sequence.

In certain embodiments the wild-type or mutant status of the ERa gene is determined by the presence or absence of non-synonymous nucleic acid variations in the genes. Observed non-synonymous variations corresponding to known common

polymorphisms with no annotated functional effects do not contribute to a gene status of mutant.

Other variations in the ERa gene that signify mutant status include splice site variations that decrease recognition of an intron/exon junction during processing of pre- mRNA to mRNA. This can result in exon skipping or the inclusion of normally intronic sequence in spliced mRNA (intron retention or utilization of cryptic splice junctions). This can, in turn, result in the production of aberrant protein with insertions and/or deletions relative to the normal protein. Thus, in other embodiments, the gene has a mutant status if there is a variant that alters splice site recognition sequence at an intron/exon junction.

For ESR1, reference sequences are available for the gene (GenBank accession number: NG 008493), mRNA (GenBank accession number: NM 000125), and protein (GenBank accession number: NP 000116 or Swiss-Prot accession: P03372). A person of skill in the art will be able to determine the ESR1 gene status, i.e. whether a particular ESRlgene is wild type or mutant, based on comparison of DNA or protein sequence with wild type.

It will be apparent that the gene and mRNA sequences disclosed for ERa gene are representative sequences. In normal individuals there are two copies of each gene, a maternal and paternal copy, which will likely have some sequence differences, moreover within a population there will exist numerous allelic variants of the gene sequence. Other sequences regarded as wild type include those that possess one or more synonymous changes to the nucleic acid sequence (which changes do not alter the encoded protein sequence), non-synonymous common polymorphisms (e.g. germ-line polymorphisms) which alter the protein sequence but do not affect protein function, and intronic non-splice- site sequence changes.

There are numerous techniques available to the person skilled in the art to determine the gene status of ERa. The gene status can be determined by determination of the nucleic acid sequence. This could be via direct sequencing of the full-length gene or analysis of specific sites within the gene, e.g. commonly mutated sites.

According to one aspect of the specification there is provided a method for selecting a patient for treatment with a compound of Formula (I), the method comprising providing a tumour cell containing sample from a patient; determining whether the ERa gene in the patient's tumour cell containing sample is wild type or mutant; and selecting a patient for treatment with a compound of Formula (I), based thereon.

The method may include or exclude the actual patient sample isolation step. Thus, according to one aspect of the specification there is provided a method for selecting a patient for treatment with a compound of Formula (I), the method comprising determining whether the ERa gene in a tumour cell containing sample previously isolated from the patient is wild type or mutant; and selecting a patient for treatment with a compound of Formula (I), based thereon. In one embodiment, the patient is selected for treatment with a compound of Formula (I), if the tumour cell DNA has a mutant ERa gene. In other embodiments, a patient whose tumour cell DNA possesses a wild type ERa gene is selected for treatment with a compound of Formula (I).

In another aspect, the specification provides a method of treating a patient suffering from cancer comprising: determining the mutant or wild type status of the ERa gene in the patient's tumour cells and if the ERa gene is mutant, administering to the patient an effective amount of a compound of Formula (I).

According to another aspect of the specification there is provided the use of a compound of Formula (I), or a pharmaceutically acceptable salt thereof to treat a cancer patient whose tumour cells have been identified as possessing a mutant ERa gene.

According to another aspect of the specification there is provided a compound of Formula (I), or a pharmaceutically acceptable salt thereof for use in treating cancers with tumour cells identified as harbouring mutant ERa gene.

According to another aspect of the specification there is provided a method of treating cancers with tumour cells identified as harbouring mutant ERa gene comprising administering an effective amount of a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

For all the aspects above, mutant forms of ERa determined/identified are at all positions across the gene.

For all the aspects above, using tumours such as breast cancer as an example, particular mutant forms of ERa determined/identified are those at positions Ser463Pro, Val543Glu, Leu536Arg, Tyr537Ser, Tyr537Asn and Asp538Gly.

Examples

The specification will now be illustrated in the following Examples in which, generally:

(i) operations were carried out at ambient temperature, i.e. in the range 17 to 25 °C and under an atmosphere of an inert gas such as nitrogen unless otherwise stated;

(ii) evaporations were carried out by rotary evaporation or utilising Genevac equipment or Biotage vlO evaporator in vacuo and work-up procedures were carried out after removal of residual solids by filtration; (iii) flash chromatography purifications were performed on an automated Teledyne Isco CombiFlash® Rf or Teledyne Isco CombiFlash® Companion® using prepacked RediSep Rf Gold™ Silica Columns (20-40 μιη, spherical particles), GraceResolv™ Cartridges (Davisil® silica) or Silicycle cartridges (40 - 63 μιη).

(iv) preparative chromatography was performed on a Gilson prep HPLC instrument with UV collection or via supercritical fluid chromatography performed on a Waters Prep 100 SFC-MS instrument with MS- and UV- triggered collection or a Thar MultiGram III SFC instrument with UV collection;

(v) chiral preparative chromatography was performed on a Gilson instrument with UV collection (233 injector / fraction collector, 333 & 334 pumps, 155 UV detector) or a Varian Prep Star instrument (2 x SD1 pumps, 325 UV detector, 701 fraction collector) pump running with Gilson 305 injection;

(vi) yields, where present, are not necessarily the maximum attainable;

(vii) in general, the structures of end-products of the Formula (I) were confirmed by nuclear magnetic resonance (NMR) spectroscopy; NMR chemical shift values were measured on the delta scale [proton magnetic resonance spectra were determined using a Bruker Avance 500 (500 MHz) or Bruker Avance 400 (400 MHz) instrument];

measurements were taken at ambient temperature unless otherwise specified; the following abbreviations have been used: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; dd, doublet of doublets; ddd, doublet of doublet of doublet; dt, doublet of triplets; bs, broad signal

(viii) in general, end-products of the Formula (I) were also characterised by mass spectroscopy following liquid chromatography (LCMS or UPLC); UPLC was carried out using a Waters UPLC fitted with Waters SQ mass spectrometer (Column temp 40, UV = 220-300nm, Mass Spec = ESI with positive/negative switching) at a flow rate of lml/min using a solvent system of 97% A + 3% B to 3% A to 97% B over 1.50mins (total runtime with equilibration back to starting conditions etc 1.70min), where A = 0.1% formic acid in water (for acid work) or 0.1% ammonia in water (for base work) B = acetonitrile. For acid analysis the column used was Waters Acquity HSS T3 1.8μιη 2.1 x50 mm, for base analysis the column used was Waters Acquity BEH 1.7μιη 2.1x50mm; LCMS was carried out using a Waters Alliance HT (2795) fitted with a Waters ZQ ESCi mass spectrometer and a Phenomenex Gemini -NX (50x2. lmm 5μιη) column at a flow rate of 1. lml/min 95%A to 95%B over 4 min with a 0.5 min hold. The modifier is kept at a constant 5% C (50:50 acetonitrile: water 0.1% formic acid) or D (50:50 acetonitrile: water 0.1% ammonium hydroxide (0.88 SG) depending on whether it is an acidic or basic method.

(ix) ion exchange purification was generally performed using a SCX-2 (Biotage, Propylsulfonic acid functionalized silica. Manufactured using a trifunctional silane. Non end-capped) cartridge.

(x) intermediate purity was assessed by thin layer chromatographic, mass spectral, HPLC (high performance liquid chromatography) and/or NMR analysis;

(xi) RockPhos 3^rd Generation Precatalyst was sourced from Strem Chemicals Inc. and from Sigma-Aldrich.

(xii) the following abbreviations have been used:-

AcOH acetic acid

aq. aqueous

n-BuLi n-butyl lithium

tBuOH tert-butanol

Brettphos 2-(dicyclohexylphosphino)3 ,6-dimethoxy-

2',4',6'-triisopropyl- 1 , 1 '-biphenyl

CDC deutero-chloroform

Cone. concentrated

DCM dichloromethane

DEAD diethylazodicarboxylate

DIPEA diisopropylethylamine

DMA NN-dimethylacetamide

DMAP dimethylaminopyridine

DME dimethoxyethane

DMF A ,N-dimethylformamide

DMSO dimethyl sulphoxide

EtOH ethanol

EtOAc ethyl acetate

HATU l-[bis(dimethylamino)methylene]-lH-l,2,3- triazolo[4,5-£]pyridinium 3-oxid

hexafluorophosphate HPLC high performance liquid chromatography IPA isopropyl alcohol

MeCN acetonitrile

MeOH methanol

RockPhos 3rd Generation [(2-Di-tert-butylphosphino-3-methoxy-6- Precatalyst methyl-2',4',6'-triisopropyl-l , 1 '-biphenyl)-2-

(2-aminobiphenyl)]palladium(II)

methanesulfonate

rt/PvT room temperature

Ruphos 2-dicyclohexylphosphino -2 ', 6 '- diisopropoxybiphenyl

sat. saturated

SFC supercritical fluid chromatography sol. solution

TBDMS tert-butyldimethylsilyl

THF tetrahydrofuran

Example 1

(6S,8R)-7- -fluoro-2-methylpropyl)-6-(4-q-(3-(fluoromethyl)azetidin-l-yl)ethoxy)-2- methoxyphenyl)-8-methyl-6._l7._l8._l9-tetrahvdro-3H-pyrrolo[3._l2- 1iso uinoline

Triphenylphosphine (52.2 mg, 0.20 mmol) was added to a solution of tert-butyl tra/?5-7-(2- fluoro-2-methylpropyl)-6-(4-hydroxy-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H- pyrrolo[3,2-/]isoquinoline-3-carboxylate (48 mg, 0.10 mmol) and 2-(3- (fluoromethyl)azetidin-l-yl)ethan-l-ol (16 mg, 0.12 mmol) in THF (1 mL). The reaction was cooled to 0 °C, and then DEAD in toluene (40 wt%; 0.079 mL, 0.20 mmol) was added dropwise. The reaction was then warmed to room temperature and stirred under these conditions for 20 hours. The reaction was then concentrated under reduced pressure, and the resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in hexanes followed by isocratic 20% MeOH in DCM, to afford tert-butyl tra/75-7-(2-fluoro-2-methylpropyl)-6-(4-(2-(3-(fluoromethyl)azetidin-l-yl)ethoxy)-2- methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrrolo[3,2_: ]isoquinoline-3-carboxylate (120 mg) contaminated with triphenylphosphine as an oil This material was dissolved in DCM (0.5 mL) and cooled to 0 °C. Then trifluoroacetic acid (0.50 mL) was added dropwise. The reaction was allowed to warm to room temperature and was stirred for 1 hour under these conditions. The reaction was concentrated under reduced pressure and the resulting residue was purified by preparative HPLC (Column: Xbridge Pheny; Length: 150 mm; Diameter: 19 mm, 5 μιη; Flow Rate: 20 mL/min) eluting with 60 to 80% acetonitrile in water containing 0.2% NH4OH (pH 10). Product fractions were combined and concentrated under reduced pressure, and the resulting residue was purified by preparative SFC (Column: Chiralpak AD; Length: 250 mm; Diameter: 21.2 mm; 5 μιη; Flow Rate: 75 mL/min; Outlet pressure: 100 bar; Column Temperature: 40 °C) eluting with isocratic (20% ethanol containing 0.2% NH4OH) in CO2. This afforded the title compound (5 mg, 10%) as a dry film. ^!H NMR (300MHz, DMSO-de) 0.97 (3H, d), 1.16 - 1.32 (6H, m), 2.17 - 2.36 (1H, m), 2.62 - 2.84 (5H, m), 2.95 - 3.13 (3H, m), 3.26 - 3.34 (2H, m), 3.48 - 3.63 (1H, m), 3.81 - 3.89 (5H, m), 4.50 (2H, dd), 5.26 (1H, s), 6.28 (1H, dd), 6.37 - 6.43 (2H, m), 6.53 (1H, d), 6.63 (1H, d), 7.04 (1H, d), 7.27 (1H, t), 10.92 - 10.98 (1H, m). m/z: ES+ [M+H]+ 498.

Procedures used to prepare the starting material tert- vXy\ tra/?5-7-(2-fluoro-2- methylpropyl)-6-(4-hydroxy-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H- pyiTolo[3,2-/]isoquinoline-3-carboxylate are described below. l-(lH-indol-4-yl)propan-2-one

Prop-l-en-2-yl acetate (4.49 mL, 40.8 mmol), tributyl(methoxy)stannane (11.8 mL, 40.8 mmol), tri-o-tolylphosphine (0.621 g, 2.04 mmol), and palladium(II) acetate (0.229 g, 1.02 mmol) were added to a solution of 4-bromo-lH-indole (2.56 mL, 20.4 mmol) in toluene (120 mL). The reaction was stirred at 100 °C for 6 hrs and then quenched by addition of a solution of 15.5 g KF in 50 mL H₂0. After 10 minutes, the resulting suspension was filtered through a pad of Celite®. The layers were separated, and the organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was adsorbed onto silica and purified by flash silica chromatography, elution gradient 0 to 80% ethyl acetate in hexanes. Product fractions were concentrated under reduced pressure and the resulting residue was triturated with 1 : 1 ether:hexanes to afford l-(lH-indol-4-yl)propan-2-one (3.24 g, 92%) as a solid. ^!H NMR (300 MHz, DMSO-de, 27 °C) 2.06 (3H, s), 3.90 (2H, s), 6.36 - 6.43 (1H, m), 6.83 (1H, d), 6.97 - 7.07 (1H, m), 7.24 - 7.35 (2H, m), 11.09, (1H, br s). l-( lH-indol-4-vDpropan-2-amine, formic acid salt

Ammonium formate (11.85 g, 187.9 mmol) and palladium on carbon (5 wt%; 2.0 g, 0.94 mmol) were added sequentially to l-(lH-indol-4-yl)propan-2-one (2.17 g, 12.5 mmol) dissolved in MeOH (60 mL). The reaction was stirred at room temperature under nitrogen for 18 hours. The reaction was filtered through a pad of Celite®, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase HPLC (Column: CSH CI 8; Length: 100 mm; Diameter: 30 mm; 5 μιη; Flow rate: Flow rate: 40 mL/min) eluting with 1 to 17% acetonitrile in water containing 0.1% formic acid (pH 3) over 4 minutes. Product fractions were combined and concentrated under reduced pressure to afford l-(lH-indol-4-yl)propan-2-amine (2.35 g, 85%) as a formic acid salt and a gum. ^!H NMR (300MHz, DMSO-de, 27 °C) 1.05 (3H, d), 2.81 (1H, dd), 3.06 - 3.15 (1H, m), 3.32 - 3.46 (1H, m), 6.51 - 6.53 (1H, m), 6.80 (1H, d), 7.01 (1H, t), 7.27 (1H, d), 7.32 (1H, t), 8.39 (1H, br s), 11.11 (1H, br s). N⁺H₃ not observed, m/z: ES+ [M+H]+ 175.

Alternative preparation of l-(lH-indol-4-yr)propan-2-amine, free base

Ammonium formate (27.3 g, 433 mmol) and palladium on carbon (10 wt%; 2.30 g, 2.16 mmol) were added sequentially to l-(lH-indol-4-yl)propan-2-one (5 g, 28.9 mmol) dissolved in MeOH (100 mL). The reaction was stirred at room temperature under nitrogen for 19 hours. The reaction was filtered through a pad of Celite®, and the filtrate was concentrated under reduced pressure. The resulting residue was suspended in saturated aqueous K2CO3 and extracted with EtOAc. The organic layer was washed with saturated aqueous sodium chloride, dried over sodium sulfate, filtered, and concentrated under reduced pressure to afford l-(lH-indol-4-yl)propan-2-amine (4.90 g, 97%) as a free base and a solid. ^lH NMR (300MHz, DMSO-de, 27 °C) 0.97 (3H, d), 1.33 (2H, br s), 2.66 - 2.85 (2H, m), 3.15 (1H, sxt), 6.45 - 6.50 (1H, m), 6.78 (1H, d), 6.99 (1H, dd), 7.23 (1H, d), 7.27 - 7.30 (1H, m), 11.01 (lH, br s). m/z: ES+ [M+H]+ 175. V-fl-flH-indol-4-yl)propan-2-yl)-4-chloro-2-methoxybenzamide

l-(lH-Indol-4-yl)propan-2-amine, formic acid salt (2.35 g, 10.7 mmol) dissolved in 50%> methanol in DCM (40 mL) was added to a suspension of MP -carbonate resin (20 g, 21.3 mmol carbonate at 1.08 mmol/g) in 50% methanol in DCM (40 mL). After stirring for one hour, the mixture was filtered, and the filtrate concentrated under reduced pressure to afford a white solid (1.35 g). This solid was dissolved in DMF (22 mL) and 4-chloro-2- methoxybenzoic acid (1.59 g, 8.52 mmol), HATU (3.24 g, 8.52 mmol), and TEA (2.16 mL, 15.5 mmol) were added. The reaction was stirred at room temperature for 2 hours and was then quenched with water and extracted with EtOAc. The organic layer was washed with saturated aqueous sodium chloride, dried over sodium sulfate, and filtered. The filtrate was adsorbed onto silica gel and purified by flash silica chromatography, elution gradient 0 to 80% ethyl acetate in hexanes, to afford N-(l-(lH-indol-4-yl)propan-2-yl)-4- chloro-2-methoxybenzamide (2.28 g, 62.4%) as a foam solid. ^!H NMR (300MHz, DMSO- <k, 27 °C) 1.14 (3H, d), 2.91 (1H, dd), 3.17 (1H, dd), 3.85 (3H, s), 4.36 (1H, dt), 6.62 - 6.66 (1H, m), 6.85 (1H, d), 6.99 - 7.10 (2H, m), 7.20 (1H, d), 7.26 (1H, d), 7.29 - 7.33 (1H, m), 7.70 (1H, d), 7.99 (1H, d), 11.04 (1H, br s).

6-f4-chloro-2-methoxyphenyl)-8-methyl-8._l9-dihvdro-3H-pyrrolo[3._l2- 1iso uinoline

N-(l-(lH-Indol-4-yl)propan-2-yl)-4-chloro-2-methoxybenzamide (0.815 g, 2.38 mmol) was dissolved in phosphorus oxychloride (4.43 mL, 47.6 mmol) and subjected to microwave conditions (300W, 110 °C) for 1 hour. The reaction was then concentrated under reduced pressure. The resulting residue was dissolved in EtOAc, washed with saturated aqueous sodium bicarbonate, dried over sodium sulfate, filtered, and

concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 30 to 100% EtOAc in hexanes, to afford 6-(4-chloro-2- methoxyphenyl)-8-methyl-8,9-dihydro-3H-pyrrolo[3,2_: ]isoquinoline (558 mg, 72.3%) as a yellow film. ^!H NMR (300MHz, DMSO-de, 27 °C) 1.31 - 1.53 (3H, br m), 2.59 - 2.79 (1H, m), 3.03 - 3.22 (1H, m), 3.53 - 3.84 (4H, br m), 6.52 - 6.72 (2H, m), 7.06 - 7.35 (4H, m), 7.40 - 7.47 (1H, m), 11.03 - 11.63 (lH, br s). m/z: ES+ [M+H]+ 325.

^TO¾s-6-f4-chloro-2-methoxyphenyl)-8-methyl-6._l7._l8._l9-tetrahvdro-3H-pyrrolo[3._l2-

/lisoquinoline

racemic 6-(4-Chloro-2-methoxyphenyl)-8-methyl-8,9-dm^ (0.793 g, 2.44 mmol) was suspended in THF (30 mL) and cooled to -78 °C. Then lithium aluminium hydride in THF (2 M; 9.03 mL, 18.1 mmol) was added dropwise, slowly, followed by dropwise addition of trimethylaluminium in toluene (2 M; 9.03 mL, 18.1 mmol). The reaction was maintained at -78 °C for 30 min and then warmed to -45°C. After one hour, the reaction was warmed to -25 °C. After 1 hour, the reaction was warmed to 0 °C. After stirring under these conditions for 3.5 hours, the reaction was cooled to -45 °C and quenched by sequential addition of water (700 μί), 15% NaOH in water (700 μί), and water (2.1 mL). The resulting suspension was diluted with EtOAc and allowed to warm to room temperature. After 15 minutes, the mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was adsorbed onto silica and purified by flash silica chromatography, elution gradient 0 to 20% methanol in ethyl acetate, to afford tra/?5-6-(4-chloro-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H- pyrrolo[3,2- Jisoquinoline (218 mg, 27.3%) as a foam solid. ^!H NMR (300MHz, DMSO- <k, 27 °C) 1.07 (3H, d), 1.99 - 2.25 (1H, br m), 2.82 - 3.01 (2H, m), 3.93 (3H, s), 5.49 (1H, s), 6.36 (1H, d), 6.41 (1H, t), 6.51 (1H, d), 6.76 (1H, dd), 7.10 (1H, d), 7.13 (1H, d), 7.30 (1H, t), 11.02 (1H, br s). One hydrogen obscured by DMSO. m/z: ES+ [M+H]+ 327.

^TO¾s-6-f4-chloro-2-methoxyphenyl)-7-f2-fluoro-2-methylpropyl)-8-methyl-6.,7._l8._l9- tetrahvdro-3H-pyrrolo[3,2- 1isoquinoline

racemic

2-Fluoro-2-methylpropyl trifluoromethanesulfonate (444 mg, 1.98 mmol) and DIPEA (0.427 mL, 2.45 mmol) were added sequentially to a solution of tra/?5-6-(4-chloro-2- methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrrolo[3,2_: ]isoquinoline (216 mg, 0.66 mmol) in dioxane (6 mL). The reaction was then heated at 60 °C for 4 days. The reaction was then cooled, concentrated under reduced pressure, and the resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% EtOAc in hexanes, to afford tra/75-6-(4-chloro-2-methoxyphenyl)-7-(2-f uoro-2-methylpropyl)-8-methyl-6,7,8,9- tetrahydro-3H-pyiTolo[3,2-/]isoquinoline (185 mg, 69.8%) as a light yellow foam solid. ^!H NMR (300MHz, DMSO-de, 27 °C) 0.97 (3H, d), 1.15 - 1.30 (6H, m), 2.16 - 2.35 (1H, m), 2.70 - 2.86 (2H, m), 3.10 (1H, br dd), 3.49 - 3.57 (1H, m), 3.90 (3H, s), 5.31 (1H, s), 6.37 - 6.43 (2H, m), 6.76 - 6.82 (2H, m), 7.03 - 7.09 (2H, m), 7.28 (1H, t), 10.98 (1H, br s). m/z: ES+ [M+H]+ 401. fe -butyl ^m¾s-6-f4-chloro-2-methoxyphenyl)-7-f2-fluoro-2-methylpropyl)-8-methyl- 6,7._l8._l9-tetrahv(iro-3H-pyrrolo[3._l2-f|iso uinoline-3-carboxylate

racemic

Di-tert-butyldicarbonate (0.107 mL, 0.46 mmol) was added to a solution of tra/?5-6-(4- chloro-2-methoxyphenyl)-7-(2-fluoro-2-methylpropyl)-8-methyl-6,7,8,9-tetrahydro-3H- pyiTolo[3,2-/]isoquinoline (185 mg, 0.46 mmol) and DMAP (67.6 mg, 0.55 mmol) in acetonitrile (2 mL). The reaction was stirred at room temperature for 2 hours. The reaction was then diluted with EtOAc, washed with water, and the organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes, to afford tert-butyl tra/75-6-(4-chloro-2-methoxyphenyl)-7-(2-fluoro-2-methylpropyl)-8- methyl-6,7,8,9-tetrahydro-3H-pyrrolo[3,2-fJisoquinoline-3-carboxylate (188 mg, 81%) as a yellow film. ^!H NMR (300MHz, DMSO-de, 27 °C) 0.96 (3H, d), 1.09 - 1.31 (6H, m), 1.60 (9H, s), 2.13 - 2.30 (1H, m), 2.74 - 2.86 (2H, m), 3.15 (1H, br dd), 3.52 - 3.64 (1H, m),

3.89 (3H, s), 5.32 (1H, s), 6.63 (1H, d), 6.74 (1H, d), 6.77 - 6.85 (2H, m), 7.10 (1H, s), 7.66 (1H, d), 7.70 (1H, d). m/z: ES+ [M+H]+ 501. fe -butyl ^m¾s-7-f2-fluoro-2-methylpropyl)-6-f4-hydroxy-2-methoxyphenyl)-8- methyl-6._l7._l8._l9-tetrahvdro-3H-pyrrolo[3._l2- liso uinoline-3-carboxylate

7¾rt-butyl tran5-6-(4-chloro-2-methoxyphenyl)-7-(2-fiuoro-2-methylpropyl)-8-methyl- 6,7,8,9-tetrahydro-3H-pyrrolo[3,2_: ]isoquinoline-3-carboxylate (188 mg, 0.38 mmol), Pd₂dba3 (7 mg, 8 μιηοΐ), di-tert-butyl(2',4',6'-triisopropyl-3,4,5,6-tetramethyl-[l, - biphenyl]-2-yl)phosphane (14 mg, 0.030 mmol), and KOH (42.1 mg, 0.75 mmol) were added to a 25 mL round-bottom flask, and the flask was evacuated and backfilled with nitrogen (3x). Then 1,4-dioxane (0.50 mL) and water (0.5 mL) were added, and the reaction was placed in an oil bath that had been preheated to 90 °C. The reaction was maintained under these conditions for 3 hours. The reaction was then cooled to room temperature and carefully neutralized with aqueous HCl (IN). The mixture was extracted with EtOAc, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in hexanes, to afford tert-butyl tra/?5-7-(2-fluoro-2-methylpropyl)-6-(4- hydroxy-2-methoxyphenyl)-8-methyl-6,7,8,9-tetrahydro-3H-pyrrolo[3,2-fJisoquinoline-3- carboxylate (48 mg, 27%) as a yellow film. ^!H NMR (300MHz, DMSO-de, 27 °C) 0.96 (3H, d), 1.11 - 1.26 (6H, m), 1.54 - 1.67 (9H, m), 2.12 - 2.37 (IH, m), 2.69 - 2.82 (2H, m), 3.12 (IH, br dd), 3.53 - 3.65 (IH, m), 3.72 - 3.87 (3H, m), 5.23 (IH, s), 6.16 (IH, dd), 6.41 (IH, d), 6.55 (IH, d), 6.62 (IH, d), 6.72 (IH, d), 7.63 (IH, d), 7.68 (IH, d), 9.28 (IH, s). m/z: ES+ [M+H]+ 483.

Example 2:

V-a-(3-fluoropropyl)azetidin-3-yl)-6- 6S,8R)-8-methyl-7-q,2,2-trifluoroethyl)-

6._l7._l8._l9-tetrahvdro-3H-pyrrolo[3._l2- liso uinolin-6-yl)pyridin-3-amine

l-(3-Fluoropropyl)azetidin-3-amine (13 mg, 0.080 mmol) was added to a red-orange solution of (65^,,8i?)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6, 7,8,9- tetrahydro-3H-pyrrolo[3,2-/]isoquinoline (30 mg, 0.070 mmol) in 1,4-dioxane (0.35 mL). Nitrogen was bubbled through the solution for 5 minutes. Then solid sodium tert-butoxide (27 mg, 0.28 mmol) was added followed by BrettPhos 3rd Generation Precatalyst (3.2 mg, 3.5 μιηοΐ), and the reaction was heated at 50 °C for 1.5 hours.

In a separate flask l-(3-fluoropropyl)azetidin-3-amine (72 mg, 0.43 mmol) was added to a red-orange solution of (65',8i?)-6-(5-bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)- 6,7,8,9-tetrahydro-3H-pyrrolo[3,2_: ]isoquinoline (167 mg, 0.39 mmol) in 1,4-dioxane (2 mL). Nitrogen was bubbled through the solution for 5 minutes. Solid sodium tert-butoxide (151 mg, 1.57 mmol) was added followed by BrettPhos 3rd Generation Precatalyst (18 mg, 0.020 mmol), and reaction was heated at 50 °C for 2.5 hours.

The contents of both reactions were combined and concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% methanol in DCM, to afford an amber foam solid (216 mg). This material was further purified by preparative SFC (Column: (S,S) Whelk-Ol, Length: 250 mm, Diameter: 30 mm, 5 μιη; Flow rate: 120 mL/min), eluting with 30% (0.2% NH₄OH in methanol) in CO2, as a yellow solid (175 mg). This solid was repurified by flash silica chromatography, elution gradient 0 to 40% methanol in DCM. Product fractions were concentrated under reduced pressure to afford the title compound (116 mg, 62.0%>) as a pale yellow foam. ^!Η NMR (300MHz, DMSO-de, 27 °C) 1.07 (3H, d), 1.55 - 1.75 (2H, m), 2.42 - 2.49 (2H, m), 2.69 - 2.81 (3H, m), 2.88 - 3.05 (2H, m), 3.42 - 3.56 (2H, m), 3.58 - 3.67 (2H, m), 3.87 - 4.00 (1H, m), 4.45 (2H, dt), 4.91 (1H, s), 6.18 (1H, d), 6.39 - 6.43 (1H, m), 6.54 (1H, d), 6.78 - 6.83 (1H, m), 6.85 - 6.93 (1H, m), 7.09 (1H, d), 7.29 (1H, t), 7.75 (1H, d), 11.00 (1H, s). m/z: ES+ [M+H]+ 503.

Procedures used to prepare the starting material (65',8i?)-6-(5-bromopyridin-2-yl)-8- methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrrolo[3,2- Jisoquinoline are described below. 2-f2-bromo-6-nitrophenyl)ethan-l-ol

Potassium hydroxide (5.71 g, 101.8 mmol) was added to a stirring solution of l-bromo-2- methyl-3 -nitrobenzene (20 g, 93 mmol) and paraformaldehyde (2.78 g, 92.6 mmol) in N,N- dimethylacetamide (200 mL) at 20 °C. The resulting mixture was stirred at room temperature for 45 minutes and then diluted with ethyl acetate (200 mL) and washed with water (3x) and saturated aqueous sodium chloride. The organic layer was concentrated under reduced pressure, and the resulting residue was purified by flash silica

chromatography, elution gradient 0 to 60% EtOAc in hexanes. Product fractions were concentrated under reduced pressure to afford 2-(2-bromo-6-nitrophenyl)ethan-l-ol (16 g,

72%) as a yellow solid. ^lH NMR (300MHz, DMSO-de, 27 °C) 3.09 (2H, t), 3.51 - 3.66 (2H, td), 4.89 (1H, t), 7.41 (1H, t), 7.86 (1H, dd), 7.96 (1H, dd). m/z: ES+ [M-OH+H]+

227.

2-f2-amino-6-bromophenyl)ethan-l-ol

Ammonium formate (36.6 g, 581 mmol) was added slowly, portionwise to a stirred slurry of 2-(2-bromo-6-nitrophenyl)ethan-l-ol (14.3 g, 58.1 mmol) and zinc (38.0 g, 581 mmol) in MeOH (300 mL) at 20 °C over a period of 10 minutes. The resulting mixture was stirred at room temperature for one hour and then filtered through Celite® with a methanol wash. The filtrate was concentrated under reduced pressure, and the resulting residue was taken up in ethyl acetate (150 mL). The mixture was washed with water (150 mL), the layers were separated, and the aqueous layer was extracted with ethyl acetate (2 x 100 mL). The combined organic layers were dried over Na₂S04, filtered, and concentrated under reduced pressure to afford 2-(2-amino-6-bromophenyl)ethan-l-ol (12.10 g, 96 %) as light yellow solid. ^!H NMR (300MHz, DMSO-de, 27 °C) 2.82 (2H, t), 3.48 - 3.57 (2H, 4.74 (1H, t), 5.18 (2H, s), 6.62 (1H, dd), 6.70 - 6.86 (2H, m). m/z: ES+ [M+H]+ 216.

2-f2-bromo-6-f2,5-dimethyl-lH-pyrrol-l-yl)phenyl)ethan-l-ol

pTsOH monohydrate (1.02 g, 5.37 mmol) was added to a stirring solution of 2-(2-amino-6- bromophenyl)ethan-l-ol (11.6 g, 53.7 mmol) and hexane-2,5-dione (6.95 mL, 59.1 mmol) in toluene (127 mL). The resulting solution was stirred at 100 °C for one hour. The reaction was then concentrated under reduced pressure, and the resulting residue was taken up in water (200 mL) and saturated aqueous sodium bicarbonate (50 mL). The mixture was extracted with ethyl acetate (2 x 100 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% EtOAc in hexanes. Product fractions were concentrated under reduced pressure to afford 2-(2- bromo-6-(2,5-dimethyl-lH-pyrrol-l-yl)phenyl)ethan-l-ol (15.5 g, 98.0%) as a yellow waxy solid. ^!H NMR (300MHz, DMSO-de, 27 °C) 1.85 (6H, s), 2.53 - 2.64 (2H, m), 3.32 - 3.48 (2H, m), 4.70 (1H, t), 5.84 (2H, s), 7.18 (1H, dd), 7.31 (1H, t), 7.73 (1H, dd). m/z: ES+ [M+H]+ 294. l-f3-bromo-2-f2-ff4-methoxybenzyl)oxy)ethyl)phenyl)-2,5-dimethyl-lH-pyrrole

Sodium hydride (60 wt% in mineral oil; 190 mg, 4.74 mmol) was added to a stirred solution of 2-(2-bromo-6-(2,5-dimethyl-lH-pyrrol-l-yl)phenyl)ethan-l-ol (930 mg, 3.16 mmol) in DMF (9 mL) at 0 °C. The resulting mixture was stirred at 0 °C for 10 minutes and then l-(chloromethyl)-4-methoxybenzene (0.601 mL, 4.43 mmol) was added followed by sodium iodide (47 mg, 0.32 mmol). The reaction was stirred at room temperature for

18 hours.

In a separate flask, sodium hydride (60 wt% in mineral oil; 2.93 g, 73.37 mmol) was added to a stirred solution of 2-(2-bromo-6-(2,5-dimethyl-lH-pyrrol-l-yl)phenyl)ethan-l-ol (13.5 g, 45.9 mmol) in DMF (82 mL) at 0 °C. The resulting mixture was stirred at 0 °C for 10 minutes and then l-(chloromethyl)-4-methoxybenzene (9.33 mL, 68.78 mmol) was added followed by sodium iodide (0.687 g, 4.59 mmol). The ice bath was removed, and the reaction was stirred under these conditions for six hours.

The separate reactions were slowly quenched with water until no gas evolution was observed. Both reaction mixtures were then combined. The resulting mixture was extracted with ethyl acetate (2x), and the combined organic layers were washed with saturated aqueous sodium chloride, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% EtOAc in hexanes. Product fractions were concentrated under reduced pressure to afford l-(3-bromo-2-(2-((4-methoxybenzyl)oxy)ethyl)phenyl)-2,5- dimethyl-lH-pyrrole (16.3 g, 80.3%) as a pale yellow oil. ^lH NMR (300MHz, DMSO-de, 27 °C) 1.81 (6H, s), 2.66 (2H, t), 3.35 (2H, t), 3.73 (3H, s), 4.25 (2H, s), 5.85 (2H, s), 6.83 - 6.92 (2H, m), 7.07 - 7.15 (2H, m), 7.18 - 7.24 (1H, m), 7.33 (1H, t),7.73 (1H, dd). m/z: ES+ [M+H]+ 414. fe^-butvnR)-a-(3- ,5-dimethyl-lH-pyrrol-l-yl)-2-q- 4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-yl)carbamate

n-Butyllithium in hexanes (2.5 M; 0.649 mL, 1.62 mmol) was added slowly over 5 minutes to a stirred solution of l-(3-bromo-2-(2-((4-methoxybenzyl)oxy)ethyl)phenyl)-2,5- dimethyl-lH-pyrrole (560 mg, 1.35 mmol) in THF (4 mL) at -78 °C. The resulting solution was stirred at -78 °C for 30 minutes. 7¾rt-butyl (4i?)-4-methyl-l,2-oxathiolane-3- carboxylate 2,2-dioxide (351 mg, 1.49 mmol) was then added in 3 equal portions to the above reaction mixture. The reaction was stirred at -78 °C for another 30 minutes and then allow to warm to room temperature. After 18 hours, the reaction was quenched with aqueous citric acid (IN), and the resulting mixture was stirred for 1 hour at room temperature. The reaction mixture was then diluted with water, extracted with ethyl acetate, and the layers were separated. The aqueous layer was extracted with ethyl acetate (2 x 100 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure.

In a separate flask, n-butyllithium in hexanes (2.5 M; 1.07 mL, 2.66 mmol) was added slowly over 5 minutes to a stirring solution of l-(3-bromo-2-(2-((4- methoxybenzyl)oxy)ethyl)phenyl)-2,5-dimethyl-lH-pyrrole (920 mg, 2.22 mmol) in THF (7 mL) at -78 °C. The resulting solution was stirred at -78 °C for 30 minutes, and then tert-butyl (4i?)-4-methyl-l,2-oxathiolane-3-carboxylate 2,2-dioxide (577 mg, 2.44 mmol) was added in 3 equal portions. The reaction was stirred at -78 °C for 30 minutes and then was allowed to warm to room temperature. After 18 hours, the reaction was quenched with aqueous citric acid (IN), and the resulting mixture was stirred at room temperature for one hour. The reaction mixture was diluted with water, extracted with ethyl acetate, and the layers were separated. The aqueous layer was extracted with ethyl acetate (2 x 100 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure.

The resulting residue was combined with that from the first reaction, and the resulting material was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. Product fractions were concentrated under reduced pressure to afford tert-butyl (i?)-(l-(3-(2,5-dimethyl-lH-pyrrol-l-yl)-2-(2-((4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-yl)carbamate (1.08 g, 61.3%) as a colorless gum. ^!H NMR (300MHz, DMSO-de, 27 °C) 1.03 (3H, d), 1.31 (9H, s), 1.80 (6H, s), 2.52 - 2.87 (4H, m), 3.21 (2H, br t), 3.69 - 3.80 (4H, m (singlet overlapped with multiplet)), 4.24 (2H, s), 5.81 (2H, s), 6.71 - 6.82 (1H, m), 6.82 - 6.89 (2H, m), 6.91 - 7.00 (1H, m), 7.07 - 7.15 (2H, m), 7.22 - 7.32 (2H, m). m/z: ES+ [M+H]+ 493. (R)-l-(3- ,5-dimethyl-lH-pyrrol-l-yl)-2-q- 4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-amine

HCl in dioxane (4 M; 1.22 mL, 4.87 mmol) was added to a stirred solution of tert-butyl (i?)-(l-(3-(2,5-dimethyl-lH-pyrrol-l-yl)-2-(2-((4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-yl)carbamate (800 mg, 1.62 mmol) in methanol (7 mL), and the resulting solution was stirred at room temperature for one hour. Additional HCl in dioxane (4 M; 0.800 mL) was added, and the reaction was stirred under these conditions for another hour. The reaction was concentrated under reduced pressure, and the resulting residue was diluted with ethyl acetate, washed with saturated aqueous sodium hydrogencarbonate, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 45% MeOH in DCM. Product fractions were concentrated under reduced pressure to afford (i?)-l-(3-(2,5-dimethyl-lH-pyrrol-l-yl)-2-(2-((4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-amine (364 mg, 57.1%) as a colorless oil. ^!H

NMR (300MHz, DMSO-de, 27 °C) 0.97 (3H, d), 1.79 (6H, s), 2.10 (2H, br s), 2.53 - 2.77 (4H, m), 3.05 (1H, sxt), 3.20 (2H, br t), 3.73 (3H, s), 4.22 (2H, s), 5.82 (2H, s), 6.79 - 6.89 (2H, m), 6.90 - 7.02 (1H, m), 7.06 - 7.16 (2H, m), 7.26 - 7.35 (2H, m). m/z: ES+ [M+H]+

393. fR)-3-f2-aminopropyl)-2-f2-ff4-methoxybenzyl)oxy)ethyl)aniline

Aqueous hydroxylamine (50 wt%; 0.302 mL, 4.94 mmol) and hydroxylamine

hydrochloride (172 mg, 2.74 mmol) were added to a stirred solution of (i?)-l-(3-(2,5- dimethyl- lH-pyrrol- 1 -yl)-2-(2-((4-methoxybenzyl)oxy)ethyl)phenyl)propan-2-amine ( 194 mg, 0.49 mmol) in EtOH (2 mL). The reaction was heated at 50 °C over for 18 hours and then concentrated under reduced pressure.

In a separate flask, aqueous hydroxylamine (50 wt%; 2.62 mL, 42.8 mmol) was added to a stirred solution of (R)- 1 -(3-(2,5-dimethyl- lH-pyrrol- 1 -yl)-2-(2-((4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-amine (1.68 g, 4.28 mmol) in EtOH (20 mL). The reaction was heated at 80 °C for 18 hours. Then hydroxylamine hydrochloride (1.49 g, 21.4 mmol) and hydroxylamine in water (50 wt%; 2.62 mL, 42.80 mmol) were added, and reaction was stirred under these conditions for an additional seven hours. The reaction was cooled and concentrated under reduced pressure.

The resulting residue was combined with that from the first reaction and then diluted with water, basified with saturated aqueous sodium hydrogencarbonate, and extracted with DCM and then ethyl acetate (2x). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by reverse phase flash chromatography (Column: RediSep Rf Gold^® Reversed- phase CI 8), elution gradient 0 to 90% acetonitrile in water conaining 0.1% NH₄OH.

Product fractions were concentrated under reduced pressure to dryness to afford (R)-3-(2- aminopropyl)-2-(2-((4-methoxybenzyl)oxy)ethyl)aniline (0.538 g, 35.8%) as a colorless oil. ^!H NMR (300MHz, DMSO-de, 27 °C) 0.96 (3H, d), 1.50 (2H, br s), 2.47 (2H, br dd), 2.71 - 2.87 (2H, m), 2.88 - 3.02 (1H, m), 3.50 (2H, br t), 3.76 (3H, s), 4.42 (2H, s), 4.75 (2H, s), 6.39 (1H, dd), 6.51 (1H, dd), 6.83 (1H, t), 6.87 - 6.95 (2H, m), 7.21 - 7.27 (2H, m).

fR)- V-fl-f3-amino-2-f2-ff4-methoxybenzyl)oxy)ethyl)phenyl)propan-2-yl)-2,2,2- trifluoroacetamide

Ethyl 2,2,2-trifluoroacetate (0.24 mL, 2.0 mmol) was added to a stirred solution of ( ?)-3- (2-aminopropyl)-2-(2-((4-methoxybenzyl)oxy)ethyl)aniline (527 mg, 1.68 mmol) and DIPEA (0.293 mL, 1.68 mmol) in MeOH (6 mL) at 20 °C, and the resulting solution was stirred at room temperature for two hours. The reaction was then concentrated under reduced pressure, and the resulting residue was purified by flash silica chromatography, elution gradient 0 to 25% MeOH in DCM. Product fractions were concentrated under reduced pressure to afford (i?)-N-(l -(3-amino-2-(2-((4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-yl)-2,2,2-trifluoroacetamide (706 mg, 103%) contaminated with a small amount of dichloromethane as a colorless gum. ^!H NMR

(300MHz, DMSO- e, 27 °C) 1.12 (3H, d), 2.68 (1H, dd), 2.73 - 2.98 (3H, m), 3.45 - 3.59 (2H, m), 3.76 (3H, s), 3.94 - 4.1 1 (1H, m), 4.42 (2H, s), 4.79 (2H, s), 6.39 (1H, dd), 6.52 (1H, dd), 6.83 (1H, t), 6.86 - 6.95 (2H, m), 7.13 - 7.37 (2H, m), 9.29 (1H, br d). m/z: ES+ [M+H]+ 41 1.

(R)-2- -«4-methoxybenzyl)oxy)ethyl)-3-(2-((2,2,2- trifluoroethyl)amino)propyl)aniline

Borane tetrahydrofuran complex in THF (1 M; 2.92 mL, 2.92 mmol) was added dropwise to a stirred solution of (i?)-N-(l-(3-amino-2-(2-((4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-yl)-2,2,2-trifluoroacetamide (200 mg, 0.49 mmol) in THF (3 mL) at 20 °C. The resulting solution was stirred at 65 °C for 5 hours and then cooled to room temperature. Methanol (3 mL) was added slowly, and the reaction was then stirred at room temperature for 15 minutes. Palladium on carbon (10 wt%; 52 mg) was added under a nitrogen atmosphere, and the reaction was stirred at 65 °C for one hour before being cooled to room temperature.

In a separate flask, borane tetrahydrofuran complex in THF (7.31 mL, 7.31 mmol) was added dropwise to a stirred solution of (i?)-N-(l-(3-amino-2-(2-((4- methoxybenzyl)oxy)ethyl)phenyl)propan-2-yl)-2,2,2-trifluoroacetamide (500 mg, 1.22 mmol) in THF (4.9 mL) at 20 °C. The resulting solution was stirred at 65 °C for 5 hours. The reaction was cooled to room temperature and quenched by slow addition of methanol (3 mL). After 15 minutes, palladium on carbon (10 wt%; 130 mg) was added, and the reaction was stirred at 65 °C for one hour before being cooled to room temperature.

Both reactions were filtered through Celite®, and the combined filtrates were concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 30% MeOH in DCM. Product fractions were concentrated under reduced pressure to afford (i?)-2-(2-((4-methoxybenzyl)oxy)ethyl)-3-(2-((2,2,2- trifluoroethyl)amino)propyl)aniline (524 mg, 77%) as a colorless gum. ^!H NMR (300MHz, DMSO-de, 27 °C) 0.90 (3H, d), 2.07 - 2.21 (1H, m), 2.27 - 2.42 (1H, m), 2.69 - 2.88 (4H, m), 3.14 - 3.27 (2H, m), 3.49 (2H, br t), 3.74 (3H, s), 4.40 (2H, s), 4.76 (2H, s), 6.38 (1H, dd), 6.49 (1H, dd), 6.82 (1H, t), 6.86 - 6.91 (2H, m), 7.20 - 7.26 (2H, m). m/z: ES+

[M+H]+ 397. aS,3R)-l-(5-bromopyridin-2-yl)-5- -q4-methoxybenzyl)oxy)ethyl)-3-methyl-2- ,2,2- trifluoroethyl)- 1 ,2,3i4-tetrahydroisoq uinolin-6-amine

Ytterbium(III) trifluoromethanesulfonate (3.13 mg, 5.04 μηιοΐ) was added to a stirring solution of (i?)-2-(2-((4-methoxybenzyl)oxy)ethyl)-3-(2-((2,2,2- trifluoroethyl)amino)propyl)aniline (100 mg, 0.25 mmol), 5-bromopicolinaldehyde (51.6 mg, 0.28 mmol), and water (0.023 mL, 1.26 mmol) in acetonitrile (2 mL). The resulting mixture was stirred at 70 °C for 45 minutes. The reaction was concentrated under reduced pressure, and the resulting residue was purified by flash silica chromatography, elution gradient 0 to 80% EtOAc in hexanes. Product fractions were concentrated under reduced pressure to afford (15',3i?)-l-(5-bromopyridin-2-yl)-5-(2-((4-methoxybenzyl)oxy)ethyl)-3- methyl-2-(2,2,2-trifluoroethyl)-l,2,3,4-tetrahydroisoquinolin-6-amine (109 mg, 77%) as a 4: 1 transxis ratio of diastereomers and a pale yellow foam. Trans: ^lH NMR (300 MHz, DMSO-de, 27 °C) 0.98 (3H, d), 2.52 - 2.61 (1H, m), 2.67 - 2.82 (3H, m), 2.83 - 2.98 (1H, m), 3.18 - 3.28 (1H, m), 3.38 - 3.54 (3H, m), 3.73 (3H, s), 4.41 (2H, s), 4.74 (2H, s), 4.83 (1H, s), 6.45 (2H, d), 6.84 - 6.90 (2H, m), 7.11 - 7.28 (3H, m), 7.94 (1H, dd), 8.56 (1H, d). m/z: ES+ [M+H]+ 564.

2- iS,3R)-6-amino-l-(5-bromopyridin-2-yl)-3-methyl-2-q,2,2-trifluoroethyl)-l,2,3,4- tetrahvdroiso uinolin-5-yl)ethan-l-ol

HCl in dioxane (4 M; 2.50 mL, 9.99 mmol) was added to a stirred solution of (^^^-^(S- bromopyridin-2-yl)-5-(2-((4-methoxybenzyl)oxy)ethyl)-3-methyl-2-(2,2,2-trifluoroethyl)- l,2,3,4-tetrahydroisoquinolin-6-amine (470 mg, 0.83 mmol) in MeOH (7 mL). The resulting solution was stirred at room temperature for 17 hours. The reaction was concentrated under reduced pressure, and the resulting residue was taken up in ethyl acetate, washed with saturated aqueous sodium hydrogencarbonate, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% DCM in MeOH.

Product fractions were concentrated under reduced pressure to afford 2-((15',3i?)-6-amino- l-(5-bromopyridin-2-yl)-3-methyl-2-(2,2,2-trifluoroethyl)-l,2,3,4-tetrahydroi

yl)ethan-l-ol (292 mg, 79%) as a 7: 1 transxis ratio of diastereomers and a pale yellow foam. ^!H NMR (300MHz, DMSO-de, 27 °C) 1.03 (3H, d), 2.54 - 2.69 (3H, m), 2.71 - 3.01 (2H, m), 3.18 - 3.29 (1H, m), 3.39 - 3.56 (3H, m), 4.66 (1H, t), 4.73 (2H, s), 4.84 (1H, s), 6.29 - 6.62 (2H, m), 7.23 (1H, d), 7.95 (1H, dd), 8.57 (1H, d). m/z: ES+ [M+H]+ 444.

(6S,8R)-6-(5-bromopyridin-2-yl)-8-methyl-7-q,2,2-trifluoroethyl)-6,7,8,9-tetrahvdro- 3H-pyrrolo [3,2- 1 isoq uinoline

Pentamcthylcyclopentadienyl iridium dich!oride (97 mg, 0. 12 mmol ) was added to a stirred mixture of 2-((15^,,3i?)-6-amino-l-(5-bromopyridin-2-yl)-3-methyl-2-(2,2,2-trifluoroethyl)- l,2,3,4-tetrahydroisoquinolin-5-yl)ethan-l-ol (360 mg, 0.81 mmol) and K2CO3 (112 mg, 0.81 mmol) in toluene (6 mL), and the resulting mixture was stirred at 100 °C for 20 hours. The reaction was then concentrated under reduced pressure, and the resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% EtOAc in hexanes. Product fractions were concentrated under reduced pressure to afford (65^*,8i?)-6-(5- bromopyridin-2-yl)-8-methyl-7-(2,2,2-trifluoroethyl)-6,7,8,9-tetrahydro-3H-pyrrolo[3,2- Jisoquinoline (275 mg, 80%>) as a 5: 1 transxis ratio of diastereomers and a brown gum. Trans: ¾ NMR (DMSO-de) 1.09 (3Η, d), 2.81 (1Η, dd), 2.94 - 3.08 (2Η, m), 3.39 - 3.49 (1Η, m), 3.49 - 3.64 (1Η, m), 5.06 (1Η, s), 6.41 - 6.45 (1Η, m), 6.60 (1Η, d), 7.13 (1Η, d), 7.25 (1Η, d), 7.31 (1Η, t), 7.95 (1Η, dd), 8.41 - 8.78 (1Η, m), 11.05 (1Η, br s). m/z: ES+ [M+H]+ 424.

The above description of illustrative embodiments is intended only to acquaint others skilled in the art with the Applicant's specification, its principles, and its practical application so that others skilled in the art may readily adapt and apply the specification in its numerous forms, as they may be best suited to the requirements of a particular use. This description and its specific examples, while indicating embodiments of this specification, are intended for purposes of illustration only. This specification, therefore, is not limited to the illustrative embodiments described in this specification, and may be variously modified. In addition, it is to be appreciated that various features of the specification that are, for clarity reasons, described in the context of separate embodiments, also may be combined to form a single embodiment. Conversely, various features of the specification that are, for brevity reasons, described in the context of a single embodiment, also may be combined to form sub -combinations thereof.

Claims

1. A compound of Formula (I) :

R³ is H, F, Me or CH₂OH;

R⁴ is F or Me;

R⁵ is H or F; and

R⁶ is H or F;

or a pharmaceutically acceptable salt thereof.

2. A compound of Formula (I), or a pharmaceutically acceptable salt thereof, as claimed in claim 1 , for use as a medicament.

3. A compound of Formula (I), or a pharmaceutically acceptable salt thereof, as claimed in claim 1 , for use in the prevention or treatment of cancer in a warm-blooded animal.

4. A method for the prevention or treatment of cancer in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt thereof, as

5 claimed in claim 1.

5. A pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, as claimed in claim 1 , and a pharmaceutically acceptable excipient.

o

6. A compound of Formula (I), or a pharmaceutically acceptable salt thereof, and an additional anti-tumour substance, for the conjoint treatment of cancer.